00:00:00GRAYSON: You have lived in Alton Bay, New Hampshire, since 1965?

BIEMANN: Yes, and this is our permanent home since 1998, after my retirement.

GRAYSON: Do you stay here in the winter?

BIEMANN: When it gets really tough in the beginning of January, we go down to the Boston area. We have a condo in Concord, Massachusetts. I use the time to go to my office at MIT [Massachusetts Institute of Technology] and try to straighten out the loose ends.

GRAYSON: I read all of the material you sent me, and I was aware of some other things from prior reading. The major points of your career, in your life, are fairly well defined in the literature that exists from the various articles that you have written. I would like to explore things that are a little more intimate and personal that you do not normally put in journal articles. My understanding is that you were born in 1926 in Austria.

BIEMANN: I was born in Innsbruck, which is in the western part of Austria. At the age of four, my family moved to the Vienna area, which is in the eastern 00:01:00part of Austria. I grew up more or less in a little town north of Vienna, Klosterneuburg, which is a bedroom town of Vienna. I went to school there.

GRAYSON: How did you become interested in science?

BIEMANN: The European system of grammar school, high school, and university is 00:02:00quite different from that in the U.S.

My father, Willibald Biemann, was a pharmacist. It was more or less self-understood that I would study pharmacy. My older sister was also a pharmacist. It was a family profession, even though my father was the first. I did not get excited about science for any one reason; it was just understood because my father was a pharmacist, which in Austria and Germany is quite different from what it is here. The social order goes a medical doctor, a 00:03:00lawyer, and then a pharmacist.

GRAYSON: Pharmacist was a higher-ranking social position.

BIEMANN: Also, a professional position. My father owned the pharmacy, which was in the house where we lived. I more or less lived in the apartment above the pharmacy. I had access to all the things, and of course, my father was involved in it. At that time, the pharmacist made many of the prescriptions from individual components.

GRAYSON: How does that compare to the gamut of medicines that would be in a 00:04:00pharmacy today, as opposed to the 1920s and 1930s?

BIEMANN: There were much fewer chemical medicines. Back then, there were many natural products--teas made from plants and extracts of plants--and some chemical things like aspirin, and antipyrin. The doctor prescribed mixtures of those on an individual basis, so the pharmacist had to measure them out, mix them up, and put them into little paper envelopes, or make them into pills. 00:05:00There were lots of chemicals around. In high school, I just turned out to be interested in chemistry, to the point that I always had an "A." I was ahead of the other kids. That is why I got into science.

Then I went to the university after World War II. My father had died by that time, and it was understood that I would study pharmacy. There was never any question. While doing so, I realized that I was very interested in chemistry. My family had lost the pharmacy at the end of the war because that part of Austria 00:06:00was under Russian control, and my mother, Margarethe Biemann, and my two sisters did not want to live there. They left to stay with a friend of the family who had a large house in a small village near Innsbruck. Although there was no longer a reason for me to study pharmacy, I did so and finished the course with a master's degree in pharmacy in 1948 from the University of Innsbruck which I had entered in 1945.

GRAYSON: The western part of Austria was not under Russian control?

BIEMANN: Austria was divided into four zones, the east was under Russian 00:07:00control, the southeast under British control, the central part was American, and the western part was under French control. You could easily travel around in the three western zones but once you entered the Russian zone, crossed the preliminary border, you had to watch out a little bit.

When I finished with my degree in pharmacy, I had become interested in organic chemistry. It was not necessary for me to take a job as a pharmacist, so I 00:08:00continued with chemistry. In the curriculum, the first four semesters for pharmacy and chemistry were practically the same, so it was relatively easy to switch without having to start from the bottom.

GRAYSON: You decided to go into chemistry, specifically organic chemistry?

BIEMANN: Yes. That was closest to my pharmacy beginning. It also was more interesting, or it seemed to be more interesting. My mathematics background and 00:09:00talents were not that great, so I could not go into physical chemistry.

GRAYSON: When did you opt to go for a Ph.D. degree?

BIEMANN: In 1948. Because of the circumstances, after the war it turned out that I was the only graduate student in organic chemistry because, at that time, girls -- now you have to say young ladies -- generally did not go into chemistry; there were lots in pharmacy, but none in chemistry. And the men of my age were still prisoners of war in various places; unfortunately some of them 00:10:00never came back, so I was the only student in organic chemistry.

GRAYSON: Did you get involved in any military activities during that period?

BIEMANN: When I was about sixteen and a half the boys in the school had to man anti-aircraft guns to free the real soldiers who were at the front lines. We 00:11:00attended school with our teachers during the day, except when an alarm sounded and we had to run out. That lasted for a year and then my term was over. After that, I had three months at home because my father had just died. My sister was running the pharmacy, but needed some help. Since I was the next in line to be a pharmacist in the family, I was allowed to help her. At that time, one had to practice two years in a pharmacy before one could study, a practice that was later changed to the old Austrian system.

GRAYSON: You actually had to work as an apprentice before you could study pharmacy.

00:12:00BIEMANN: Yes. That was the German system. At that time, Austria was part of Germany. After the war, it changed back to the Austrian system where one studied first and then did your two-year apprenticeship. That was another fortunate thing because otherwise, if I decided to do pharmacy, I would have to spend my first two years in the pharmacy. I spent three months at home, before I was drafted into the army in June of 1944. I was able to survive unscratched. I was 00:13:00in the eastern theatre, and avoided becoming a prisoner of war by, on the last day of the war, putting on civilian clothes and simply walking home. Since I was very young, and looked even younger, I had no troubles getting back to where my mother and my sisters were.

GRAYSON: When the war ended, was it common knowledge that the war was over?

BIEMANN: By that time, we were in a place north of Dresden which is pretty far into Germany so it was quite obvious that the war would be over in a very short time. In fact, it was over on that day.

00:14:00GRAYSON: You assumed that the war was over to a certain degree.

BIEMANN: Yes, before the Russians arrived.

GRAYSON: So the objective of your family was to avoid being in a Russian occupied area at all costs.

BIEMANN: Both my mother and sisters moved from Vienna to Tyrol, and I managed to get out of what then became East Germany.

GRAYSON: So then, you went to Innsbruck University and pursued a Ph.D. degree in organic chemistry. How did you select the professor you studied under?

BIEMANN: Again, I was the only student at that level at the time. There was only 00:15:00one professor and he was head of the organic chemistry department. I did not have a choice, so I worked for him. He was a pharmaceutical chemist. He spent his career before that in a pharmaceutical company in Hungary, and later on in Germany. Only at the end of the war, he somehow ended up at the University of Innsbruck as a professor.

GRAYSON: What was his name?

BIEMANN: Hermann Bretschneider. It took three years to get it all done, from 00:16:001948 to 1951. I worked quite closely with him, even before I was his graduate student when I had to make up the difference between the pharmacy and chemistry curricula. I had to do all the laboratory experiments, which were mainly synthesizing compounds. It was more or less my job to work out those experiments 00:17:00just like a teaching assistant. However, there was no curriculum in that detail and he wasn't familiar with it, so he had to start out making up a curriculum. I was his guinea pig for lots of laboratory experiments. That generated a relatively close relationship with him, and then the obvious choice was to be a graduate student under his mentorship. Once I got my PhD, I was right away appointed as an instructor.

GRAYSON: At Innsbruck?

00:18:00BIEMANN: At the University of Innsbruck. That involved taking care of the undergraduate laboratory, which had pharmacy and chemistry students combined.

GRAYSON: I want to get a feeling for what the university existence was like then because the war had just finished. There was a great deal of unrest and the normal flow of daily life was really fairly abnormal. There weren't many students at the university, or, as you say, most of the men were still in prisoner of war camps. But the university still tried to go forward.

BIEMANN: Yes. It was mostly filled with people who just graduated from high 00:19:00school and therefore hadn't served in the army. There was not a full set of students around; but everything was in turmoil and parts of the university were destroyed. Things were partly in temporary quarters, and as far as the laboratory goes, it was hard because in a chemical laboratory you have to use gas for Bunsen burners. But there was gas only available for two hours every 00:20:00day, mainly so people could cook lunch. We could only work in the laboratory for those two hours. It was also cold, during the winter of 1945. Our professor told us to bring a brick or two to the lab. He told us that when the gas came on to heat them up over the Bunsen burner, then stand on them, so that the students could at least keep warm and do the experiment. As soon as the gas went off the students could only write up what they did or do anything that didn't need any gas and then go home.

The European -- particularly the Austrian-German academic system (and to a certain extent also the French) is very different from the one in the United 00:21:00States. There is no set curriculum. The students would just have to take the required courses and then take the required exams. It was not that in mid-term the students did an exam, and at the end of the term you took an exam and that was it. The students took the exams sometimes a year later after the course. In the United States, if you know when the person is born, you can calculate in 98 percent of the cases, when he or she gets a Bachelors degree; but in Europe, that's not at all the case.

At European universities, the entering students already know what field they 00:22:00want to major in. But in Europe, it's up to the student how they progress; so many students take a long time to finish for all kinds of reasons. But after the war, everything was in reasonable disarray and everybody tried to pick up their life, and usually under quite different circumstances. People worked hard and tried to get things done. This was not only at the university but everywhere--that was how the "Germany economic miracle" after World War II came about--that people were very dedicated to get things rebuilt. Nobody worried 00:23:00about benefits or pensions. Now it's just the opposite.

I was the only student, and in a not rigorously defined system. I could either goof off, or I could get things done, and get ahead, at my own speed. I was fortunate to be in that situation, so, I could do things as fast as possible. It 00:24:00was almost automatic that after I got my Ph.D. that I would be a lecturer. I could have stayed in the academic system, which in the Austrian-German system is somewhat different from here. Namely, that if you kind of do your job, since the universities are run by the government, all the employees are government employees, so if you make a certain step that would be equivalent to getting tenure here, it was even more fixed in what you needed to do to stay on and get 00:25:00your paycheck.

GRAYSON: So I understand that they still do in Germany have this period of Habilitation?

BIEMANN: Yes. That is the equivalent of tenure here. I understand that it is now being reconsidered and changed. But I could have stayed on for that.

GRAYSON: How long would that have typically been for the Habilitation?

BIEMANN: Between five and ten years. Once when I went over to the dean's office at Innsbruck, I saw a little announcement for a program at the Massachusetts 00:26:00Institute of Technology, which I had no idea what it was, for a summer program for foreign students. I applied for it, and was accepted along with one other Austrian, there were sixty every year.

GRAYSON: Sixty throughout Europe?

BIEMANN: There were sixty at that time throughout the world that had been affected by the war.

GRAYSON: Was this for students only?

BIEMANN: It was for people who had a final degree and had a job at a university or a government research laboratory that could not come to the U.S. for an 00:27:00entire year, but could come for the summer. It was started by a group of MIT undergraduates who had been in Europe during the war, and had seen the destruction of practically everything including universities. They felt that MIT could do something by having people come over to do some of their research that they couldn't do at their home university for lack of instrumentation or facilities. I applied, and came here to MIT by boat, as was common.

GRAYSON: Was that a week to get across the Atlantic?

00:28:00BIEMANN: It took eleven days, coming and I returned on the S.S. United States, which, at that time, had the Blue Ribbon. It was the fastest passenger liner and took only four days to get back, it's a little different than today.

GRAYSON: What summer was that?

BIEMANN: In 1954. I was three years out of graduate school at that time.

GRAYSON: You were teaching organic chemistry at Innsbruck and taking care of teaching assistants during that time?

BIEMANN: I was teaching a course in analysis of pharmaceuticals, which later had some effect.

GRAYSON: What methods were you teaching in that course?

00:29:00BIEMANN: Wet chemistry, melting points, and color reactions.

GRAYSON: What did you do at MIT when you were there that summer?

BIEMANN: I worked with Professor George Buchi who received his Ph.D. at ETH [Swiss Federal Institute of Technology] in Zurich, Switzerland and was an assistant professor here at MIT. I was assigned to his laboratory because the head of the department Arthur Cope, felt that it would be easier if I was in a laboratory with somebody who could speak German. But one of the points of coming 00:30:00to the U.S. was to perfect my English, so we really didn't speak much German.

GRAYSON: You had taken English as a language during your schooling?

BIEMANN: Yes. That was automatic. I had eight years of Latin, and five years of English. I can remember more of English than Latin. But Latin helps in foreign languages, because of the grammar, it is related to the grammar of German, French, and Italian, and not so much to English, because English grammar, fortunately, is so simple.

GRAYSON: Summer is not a very long period of time, what were you able to accomplish in that time period?

BIEMANN: I worked on some structural problems, and extended my stay until the 00:31:00end of November because the visa was for six months, so I could stay for that long. I didn't have to be back by the beginning of term. It certainly was a big turning point in my career because Professor Bretschneider was the typical professor in the Austrian-German system, like he is the general and he has some lieutenants, working under him, who take care of the troops. To stray out of 00:32:00that system was not advisable and I applied for the fellowship without his support but it wasn't needed because the application didn't ask for reference letters. The application asked what I was doing and where I lived.

When I came back, he never asked me what I did, or what I learned because it was outside of the box. I could have gone towards Habilitation with his blessing, and that required - on paper -- that it would be independent research but it was also understood that I could work on some of my own ideas. In general, I had to 00:33:00work on his research, and eventually he would permit me to publish some papers under my own name without his name on it too, to satisfy the requirement. But after seeing that there was another way of academic life in the U.S., I didn't want to go that track. So, I decided to come back to the United States.

GRAYSON: You stayed at Innsbruck until the next summer?

BIEMANN: I came back to Innsbruck in the beginning of December of 1954, and I left end of September of 1955. When George Buchi found out that I was interested 00:34:00in coming back, he offered me a post-doctoral fellowship, which I accepted.

GRAYSON: You expressed an interest in coming back to the United States and then he offered you the position?

BIEMANN: Yes. Originally, there was a little twist to it because when I arrived in 1954, on 1 June, Professor Erika Cremer, the head of physical chemistry at the University of Innsbruck was on a sabbatical at MIT. She knew that I was here, and knew George Buchi. She told him that he had to promise her that he 00:35:00will not keep me here, because she wanted me to stay at the University of Innsbruck. When I left, he didn't say "If you want to come back, you always are welcome here." But I had made contacts with a well-known steroid chemist at the University of Pennsylvania in Philadelphia, at the medical school. So the steroid chemist offered me a job when I met him at ACS [American Chemical Society] meeting in New York, in the fall of 1954.

I wrote a letter to George Buchi telling him that I was thinking about coming 00:36:00back to the U.S. and what did he think? Should I take that job with Professor Ehrenstein? He said, "That would be okay. It would be a good entry into the pharmaceutical industry, but why don't you come back to MIT." Since I decided to leave Innsbruck, he wasn't bound anymore by the request from Erika Cremer. By the way, I should mention that she was the original inventor of gas chromatography. But, that is hardly known, because she did gas solid chromatography as a physical chemistry experiment to determine heats of 00:37:00adsorption. She wrote the paper in 1943, maybe 1944 and sent it in. Her publication was accepted. It was set in print and before the journal was published the publishers' building was bombed, and everything disappeared. Later, after the war, Professor Martin from the UK [United Kingdom] came to her laboratory and saw her experiment. Soon thereafter, he published his paper on gas chromatography with Dr. James. Cremer's original paper was finally printed about thirty years later in the journal Chromatographia in Germany.

00:38:00By October 1955 I was back at MIT as a post-doc with George Buchi.

GRAYSON: When you came this time, were you planning on staying in the States?

BIEMANN: Yes, I had an immigration visa, a green card; a legal immigrant.

GRAYSON: MIT was also probably in some form of turmoil as well. Obviously, it wasn't affected by the war physically, but the people's lives had been interrupted, and this was in the early 1950s. Things had probably settled down by then.

BIEMANN: Yes. That summer program started in 1947 or 1948 because those 00:39:00undergraduates who started it had come back to finish their studies. The program was financed by the Alfred P. Sloan Foundation.

GRAYSON: I was wondering where the money came from for that. It's a neat idea but the undergraduates didn't have any money.

BIEMANN: At the time when I came, in 1954, the effect of the war was done and it was an important period for MIT because after World War II the president of MIT wanted to change and modernize the institution. Until then MIT was an 00:40:00engineering school, with science more or less supporting the engineering school. All of the scientific developments generated during World War II because of the Manhattan Project; also because of the need to make synthetic rubber and improve the refining of crude oil, of course, the great thing was medicine. For example, penicillin was discovered in England a few years before. There was a big push to manufacture penicillin so that it could be made chemically and not just 00:41:00biologically for the battlefield. By the end of World War II, chemistry had a much bigger impact on everyday life than it did before. Therefore, it was important to upgrade the chemistry at the educational level.

The president of MIT appointed Professor Arthur C. Cope as a new head of the department with the mandate to put chemistry at the top. He began to hire a 00:42:00completely new group of faculty who were in the line of those new fields. He let the untenured ones go and the tenured ones just stayed and had offices and a laboratory; and of course, salary. George Buchi was one of the new hires because he was a natural products chemist, trained in Switzerland. I should mention Professor John Sheehan who was also an organic chemist and eventually developed a synthetic route to penicillin so that penicillin could be made chemically and not just bacteriologically.

00:43:00Amongst those new outcomes was to revitalize the analytical chemistry division, and he hired David Hume and Lockhart "Buck" Rogers. David Hume I think was at the Manhattan Project, which produced lots of analytical and physical chemistry and inorganic chemistry. In the process of upgrading the analytical division 00:44:00Cope wanted to have an organic chemist in there because since he was an organic chemist he felt that there had to be an organic chemist in analytical chemistry to make sure that organic chemistry were taken care of. I had taught that course in analysis of pharmaceuticals in Innsbruck and had done a good job in the project I was working on with George Buchi, so Cope felt I might be an appropriate candidate for the position and he offered it to me. I took it because I had to think of what I would do after my two years of post-doctoral 00:45:00work. Staying at MIT was a good opportunity.

GRAYSON: What year was that?

BIEMANN: That was 1 September 1957. I came back on 1 October 1955. While most organic chemists would have wanted to stay in organic chemistry, I felt that I could do just as well in analytical chemistry if I worked on the right thing.

GRAYSON: What did you do for that two-year post-doctoral period?

00:46:00BIEMANN: I worked on the synthesis of a natural product, called muscopyridine, which is a perfume component. It comes from the perfume gland of the musk deer, which lives in Mongolia and Tibet. Some funds for my post-doctoral position came from a company for which George Buchi was a consultant in Switzerland, Firmenich Company in Geneva. At that time, this compound was not anymore used but it was an interesting structure and George Buchi said it was certainly important to 00:47:00synthesize it, proof of the structure, a ten to eleven step synthesis that a graduate student of his had completed half. When I started, there was no material left to continue the remaining synthesis, so I had to repeat the graduate student's work on a larger scale and carry it through. That got me very well acquainted with physical chemical methods, particularly ultraviolet, and infrared spectroscopy.

GRAYSON: What was the state of the art, of the IR, UV at that period? It was fairly primitive still, wasn't it?

00:48:00BIEMANN: The UV spectrophotometer we used at the laboratory was still the Beckman model, where you determined point-by-point the spectrum. The earlier infrared spectrograph was the same way. When I finished the work on muscopyridine, we already had a bench top recording, little infrared machine specifically designed for qualitative, organic, infrared spectroscopy.

GRAYSON: Do you recall what that was back then?

BIEMANN: I don't remember.

GRAYSON: It had advanced to a recording bench top instrument.

00:49:00BIEMANN: Yes. It was about two feet by three feet, and maybe one foot tall. The paper was on a vertical cylinder, so it was sticking out like a smokestack.

GRAYSON: It could be a PerkinElmer [PerkinElmer, Inc]. It sounds a little bit like PerkinElmer design.

BIEMANN: The original big machine was a Baird, built exactly after a prototype that was developed either by duPont or Dow -- I don't remember. Again, another development that happened during World War II.

GRAYSON: As an instructor, that's the starting level for a tenure track at MIT 00:50:00at the time. You started doing organic synthesis.

BIEMANN: No, I could not do organic synthesis, which was the area in which I was trained. I had to look for something that was more analytical. I figured out a scheme to determine the C-terminal amino acid in a small peptide. Fred Sanger in the UK had developed a method for marking the N-terminal amino acid of small peptides, which he had used for determining the amino acid sequence of insulin; 00:51:00the first protein -- insulin -- for which a primary structure was determined. I figured if I could do the same thing with the other end of the peptide, I could simplify a rather tedious work. I planned to use a reaction, which I had developed, while still in Innsbruck, for making certain compounds that may have medical applications. I thought of using it on a micro-chemical level on small peptides.

I sent in a grant application to NIH [National Institutes of Health] that was 00:52:00funded, but in the meantime, I found out about mass spectrometry. That was by accident that was a fortunate happening. While I was still a post-doc, Firmenich, the Swiss company which had financially supported my position, was in the flavor and fragrance business and wanted to find out what happens at a conference in that field in Chicago. Instead of sending someone from Switzerland to Chicago just to attend the conference, nowadays routine, they asked George Buchi to go there and listen to the talks. He was not interested in it because 00:53:00it wasn't anything that really concerned his field. So he asked if I would go for a few days to Chicago and attend a conference and write a report. I did, and took my first airplane ride.

GRAYSON: What did you fly in? Do you remember? Was it the old DC-3?

BIEMANN: No, it was Lockheed. I listened to the talks and one of them was by Dr. Stahl from the Quartermaster Research Center in Natick, Massachusetts. He talked 00:54:00about identification of flavors using a mass spectrometer to identify the compounds. The Quartermaster Corps, that part was involved in the preparation of food rations for the armed forces, so there was a lot of dried food and reconstituting, and the important thing was that it should taste reasonably well. So they had to look at the flavor components. He looked at the volatile stuff that comes off using a mass spectrometer on simple things like methyl butyrate and butyl acetate which he then identified by matching the spectra with 00:55:00the data in the American Petroleum Institute collection. They had compiled mass spectra of all kinds of compounds, mainly hydrocarbons, but also other small, volatile molecules.

At that time, one identified mass spectra by matching them with those of known compound. So, the significance of the method didn't sink in to me right away. Later on, when I was thinking about what to do since I had been working on structure of natural products by synthesis where at each step I had to identify and make sure that what I wanted to make was what I had at the end. That 00:56:00involved a lot of qualitative analysis of the products. During the writing of the NIH proposal on the sequencing of peptides by chemical methods, I realized that mass spectrometry might be able to do that as well and better. I got the grant for the chemical method but the rules allowed me to use different methods like mass spectrometry. That started my work on mass spectrometry of peptides.

00:57:00GRAYSON: Was the conference a full week conference or couple of days?

BIEMANN: It was three days.

GRAYSON: Did this fellow at Natick mention the type of instrumentation he used?

BIEMANN: Yes. He used a CEC 21-103.

GRAYSON: Was this in the 1957-58 timeframe?

BIEMANN: It was in the spring of 1957.

GRAYSON: When this grant came through you decided that maybe mass spec has potential, but you didn't have a mass spectrometer, so what did you do?

BIEMANN: That was the problem. I went to Arthur Cope, the head of the 00:58:00department, and asked, "Why don't we have a mass spectrometer?" He said, "Because it's a big expensive instrument, that needs a full time engineer to run it and keep it running, so we don't have one." By that time I had looked into it quite a bit so I said, "No, I think I can keep it running without an electrical engineer." He said, "Okay if you promise that it won't collect dust, I promise to find some money." When he was appointed head of the department, after the war 00:59:00with the task to revamp the department, he said he needed money for that. He had at his disposal an unrestricted amount of money. Not unrestricted in dollars, but unrestricted in what he could do with it; it was completely up to him.

He used $50,000 of that money towards that instrument, and $10,000 I got from that company in Switzerland, Firmenich. The reason why he made that comment about collecting dust was that he was worried that we would buy that instrument, and it would be delivered, it wouldn't do what I wanted it to do, and therefore 01:00:00it would just sit there and would be hard to get rid of it. But fortunately, that wasn't the case.

GRAYSON: Then you purchased the 103 instrument?

BIEMANN: The 103C, from CEC, which was delivered in May of 1958.

GRAYSON: You had been working on the chemistry you needed prior to that so you would be able to use the instrument right away?

BIEMANN: Because the peptides are not volatile, I had to devise a method of chemical conversion of the peptide to something that is more volatile -- which I did by converting it to a polyamino alcohol; which is a linear molecular exactly 01:01:00like the peptide, but it was much more volatile. And of course you weren't supposed to put things like that into a mass spectrometer because the reason why the mass spectrometer existed at that time -- the commercial types -- was that the petroleum industry during World War II had to produce more and better fuels, in part, for the Air Force. The analysis of the product from crude oil to gasoline to jet fuel was very important.

Mass spectrometry was both sensitive and highly accurate so it could do that quantitative analysis, but because of the accuracy that was required, when one 01:02:00was dealing with complex mixtures the signals had to be very accurate, and very reproducible, because we had to use the intensities of the signals from standard pure compounds, as a matrix, a mathematical matrix I mean. Since you didn't want to have to run the standards each time that you did an analysis you had to rely on the standard data being very, not only accurate, but also highly reproducible over a long period of time. This required that the mass spectrometer was always in good shape. A crucial part of the spectrometer is the ion source where the 01:03:00ions are produced. It ran at high potentials, like three kilovolts, so the ion source had to be very clean. Otherwise, the potentials through electrical leakage would fluctuate and you couldn't get a good reproducible spectrum.

The rules that could not be violated were that the ion source had to be extremely clean and it had to be at a highly precise temperature, 250 degrees, plus or minus a tenth of a degree. The compounds that I wanted to put in would immediately dirty up the ion source; I mean dirty in the context of quantitative analysis. It always had to pumped out of the ion source before putting in the 01:04:00next sample. It had to be very easily pumped off and it had to be very volatile; it could not contaminate the surfaces of the ion source. Even the acceptance test, which called for an analysis of ten hydrocarbons in the range from four carbons to eight carbons, was of no use to me because that wasn't what I wanted to do. I asked them that instead of hydrocarbons of that size to use alcohols of that molecular size.

CEC, the manufacturer was very worried about that because they thought it might not work, and then I say it doesn't work and they would have to take it back. 01:05:00But I wasn't interested in a real quantitative long-lasting analysis of that kind. The installation engineer, Don DiQuasie, after he installed it, which took four weeks and two weeks to train us to run it. He said he would do the acceptance test and that it would work. When it was time to do that test he was called away on an emergency to some refinery, so he couldn't do it, and he said for me to do it.

I ran the acceptance test and was perfectly satisfied with the performance of 01:06:00the instrument. But that was one aspect that sort of helped me, except I didn't know all the rules of the games of mass spectrometry--what you do, what you don't do, and what you never should do.

As I said, it worked out quite well. Also, because one had to keep the ion source very clean, you're not supposed to clean it yourself. It needed cleaning every two or three months perhaps; so you treated the instrument with kid gloves. You had to send the ion source back to the manufacturer in Pasadena who would clean it and send it back. We needed to have two ion sources because it 01:07:00took a while to get it back. Well, we didn't want to have that problem and associated expense. After two or three times of doing that, we just took it apart ourselves, cleaned it and put it back together.

GRAYSON: Did this still have all glass inlet system, glass connections, and glass flight tube?

BIEMANN: Yes, but not the flight tube, which was metal.

GRAYSON: So every time you got into the instrument you had to have a glass blower?

BIEMANN: Yes, you had a glass blower. There was a big ball joint which connected the glass part of the inlet system to the stainless steel part, which was the flight tube, and the part the ion source was in was sealed by Apiezon wax. The 01:08:00glass blower had to come with his torch, heat up that ball joint so that the wax got soft and then we could move the entire inlet system away and get to the ion source, and take it out.

GRAYSON: If they are going to run the ion source at 250 degrees, then that ball joint has to be greased with something that's pretty non-volatile. It sounds like glue I guess.

BIEMANN: You had to heat it up quite a bit to get it flowing enough that you could take it apart. Putting it back together was again a problem. That inlet 01:09:00system could be heated to 250 degrees I think, at least 230 degrees.

GRAYSON: You had a mass spectrometer. You started in on this problem with the sequencing of the small peptides?

BIEMANN: Yes. We started on two separate projects. One was peptide sequencing which required me to work out the chemistry to convert the non-volatile peptide to a more volatile material that retained the sequence information; it retained the backbone but removed the polar groups. That took quite a bit of laboratory 01:10:00work. With the grant from NIH came a post-doc and then I got another grant from NSF that provided for another post-doc; so those two worked on that chemistry at the outset. They were both from the University of Innsbruck because that was my main connection. I wrote to two people, Sepp Seibl and Fritz Gapp, and asked if they want to join me in the U.S. They had just graduated, got their Ph.D. in organic chemistry, and were each working at a different pharmaceutical company 01:11:00in Austria. They saw that I was doing pretty well at MIT, so they agreed to come and work for me.

So I did something else, I started work on the structure of alkaloids that is a completely different field from peptide sequencing, but more closely related to my work with George Buchi, as a post-doc. I had learned a lot about alkaloids and other natural products and there was a compound available to me which was an 01:12:00indole alkaloid called sarpagine. I determined its structure which had been proposed in the literature; but people couldn't prove it because the proof by conventional means was quite complicated and time consuming. I did it by mass spectrometry, where you didn't have to make exactly the same compound that you could use for comparison, but only a similar one. That was a quick project that was successful and led me into the field of structure determination of alkaloids by mass spectrometry.

GRAYSON: You did that independently of the work of the post-docs?

BIEMANN: Yes, because it didn't need much lab work. I just needed to run the 01:13:00spectrum of the compound, then one or two chemical reactions, and the thing was done.

GRAYSON: The whole idea of determining the structure of an alkaloid by mass spectrometry, that came to you as a result of your own background. Also at that time there was some information being published about people understanding how hydrocarbons fragmented. There were some fundamental studies that were being done, and people were trying to understand why certain fragments were formed and others weren't. Did the fragmentation studies that had been done prior give you any help or insight?

BIEMANN: Yes. Those studies were done mainly on small volatile compounds of known structure, and of course, Fred McLafferty was one of those, at that time 01:14:00at Dow Chemicals [The Dow Chemical Company] who was involved with running lots of mass spectra of known compounds, and then trying to correlate the spectra with the structure. Those were quite a few experimental rules available of which bonds cleave and which don't. That information could be used to make sense out of the mass spectra of much more complicated molecules.

GRAYSON: This is probably the first time alkaloids had ever been run by a mass spec?

BIEMANN: Yes.

GRAYSON: Because most people were interested in the simpler compounds or the hydrocarbons.

BIEMANN: The idea of determining the structure of an unknown compound by mass 01:15:00spectrometry was not around. It was mainly determining the mass spectra of known compounds to characterize them, and to be able to identify them when you run across them in another situation. Because of that, you had to have the mass spectrum of the known compound. But that would never have been possible in the case of peptides. Consider the dipeptide, in which two amino acids are linked together; since there are 20 naturally occurring amino acids, this gives you 400 dipeptides that are possible. This is only the smallest one.

Then tripeptides (they are three amino acids long) would be 8,000 so you never 01:16:00could generate a library of authentic spectra to match it. You had to be able to interpret the spectrum from scratch. The same thing was true for alkaloids. There were many known ones, but nobody was interested in those anymore, once you know the structure, that's it. But at that time there were lots of alkaloids of unknown structure around because the pharmaceutical industry was looking for them for medicinal purposes. They all came from plants. Since the determination of such a structure was very complicated, tedious and time consuming, people 01:17:00published the intermediate steps of their work. They did some chemical conversions of the molecule and various reactions on it. From the outcome of some of these reactions you could tell whether it has a hydroxyl group on it or an amino group or things like this. And from the UV spectrum you could tell what aromatic substitution it had.

People published those steps and said that they think it's about this type, and the next time around that it may be this structure, or maybe that structure and now we have to prove that. There was lots of information in the literature about 01:18:00the incomplete structures. I could just read those papers and see with which one, one might be able to do it by mass spectrometry simpler, and that worked out well.

Since I needed some alkaloids of known structures, I wrote to people who had published a real complete structure and asked them to send me a sample of the compound. I need only very, very little which was important in that field. I established contacts, first with Bill Taylor [William Taylor] at Ciba Pharmaceuticals, in Summit, New Jersey. It was a Swiss company but they had a research laboratory in New Jersey. And then shortly afterwards with Norbert Neuss at the Lilly Research Laboratories in Indianapolis and we worked together. 01:19:00They had isolated many new alkaloids but could work only one after the other, the conventional way took a long time.

GRAYSON: You gave them results of your mass spec analyses and studies. Was it a reciprocal arrangement there?

BIEMANN: Yes. I got samples from them of both known ones and unknown ones, and helped them in their work by running the mass spectrum on the ones they were working on. That was a really sort of fruitful collaboration. I became a consultant to Eli Lilly, which lasted for over twenty-five years. Later many of 01:20:00those companies established their own mass spectrometry laboratories; sometimes with my help. Lilly even hired one of my post-docs, John Occolowitz. But the alkaloid work was the type where suddenly it became very fast to determine the structure of the alkaloid, particularly since they all came from tropical plants. Since the plant produces not just one alkaloid but usually a family of alkaloids with related structures, it became easy to determine their structures once you had determined the structure of one of them. You could roll up the rest 01:21:00of it relatively quickly just from the differences in their mass spectra. By the end of the 1950s there were many alkaloids around, particularly in the pharmaceutical laboratories. And by the early 1970s all of their structures were known.

GRAYSON: Am I correct in assuming that this is the first departure of mass spec into an area of chemistry? All the other applications were either in the petroleum industry, or some flavors work was being done. I could go back and 01:22:00look at the early ASMS proceedings, but I think almost everything being done was on fundamental studies. There was some ionization potential work being done. But in terms of moving the analytical capability of the instrument outside of a fairly small niche of organic chemicals this is probably one of the first departures into a different area of chemistry.

BIEMANN: It was an expansion of the analytical side of mass spectrometry into the qualitative identification of structures of natural products; structure 01:23:00determination of molecules of unknown structures. That made it part of the toolbox of the organic chemist. For the biochemist, it took quite a bit longer. Our peptide sequencing method was much more difficult to do experimentally. Also, Pehr Edman in Sweden had developed a chemical method to chip off one amino acid after the other from a large peptide or protein and determine the sequence that way. This technique was then automated and commercialized.

01:24:00Most of the peptide and protein problems in biochemistry were at that time solved by that method. Only in those situations where it did not work or could not be applied did we use mass spectrometry to solve those problems; but the mass spectrometric peptide sequencing involved complex chemistry on a very small scale so it was not easily adapted by other laboratories. My laboratory was practically the only one that used that chemistry for mass spectrometry. All that changed when Mickey Barber invented fast atom bombardment ionization.

01:25:00GRAYSON: The limiting problem was the need to get the sample into the vapor phase so that it could be ionized?

BIEMANN: The other problem was that it dealt with a very complex mixture of relatively similar molecules. In order to do a protein structure you have to degrade the protein into small pieces, into peptides. If you have a hundred amino acid long protein, which is not an unusual size, you have, theoretically 99 dipeptides, 98 tripeptides, and 97 tetrapeptides, so the mixture is very 01:26:00complex. Now that part we solved by developing the online GC-MS method because the products had to be relatively volatile to get in the mass spectrometer. So they were also sufficiently volatile to be separated by gas chromatography.

GRAYSON: You did the degradation first and then the chemistry?

BIEMANN: We did the degradation (partial hydrolysis) and the chemistry first, and then the separation afterwards. If we did it the other way around and separated the peptides first -- which was necessary before the advent of gas chromatography; -- for example Fred Sanger used paper chromatography for his insulin work -- then you would end up with up to 99 separate dipeptides, which would have to be chemically converted separately; which would be an enormous 01:27:00task. You had to do the chemistry on the mixture and then separate it and get it into the mass spectrometer. At first we did gas chromatography off-line, and then later on-line.

GRAYSON: When did you start using gas chromatography in your lab?

BIEMANN: Off-line when I started in 1958, on-line probably 1962. It was published in 1964. At first we used it on the CEC 21-110B high-resolution instrument. The first examples were on alkaloids where we separated a mixture of alkaloids from a certain plant by gas chromatography directly into the high resolution mass spectrometer. Since the alkaloids contained carbon, hydrogen, 01:28:00nitrogen and oxygen, the elemental composition -- which you could get from a high-resolution mass spectrometer -- was very helpful.

GRAYSON: You started out with the 103, a CEC instrument. It was designed for petroleum chemists but you modified or simply used it in the way you wanted to use it. Did you use their gas inlet system?

BIEMANN: Yes. Instead of expanding the sample from a glass bulb into the inlet system, we had to inject the sample into the inlet system, through a silicon 01:29:00rubber disk with a needle. We collected it from the gas chromatograph in a melting point capillary and then took it into a needle or actually we had another little adapter that we could drop the melting point capillary into and heated it to vaporize the sample into the inlet system. There were lots of experimental tricks which I learned and weren't published in a separate paper; but that's all in my 1962 book on mass spectrometry.

GRAYSON: Then did you get a second CEC instrument?

01:30:00BIEMANN: Then we first got a Bendix Time-of-Flight mass spectrometer which we used to get things directly into the ion source. When we had worked out that way of doing it we adapted it to the CEC instrument because it had much better resolution. We put the sample directly into the ion source of the CEC 21-103C through a vacuum lock.

GRAYSON: So the Bendix Time-of-Flight did have a direct insertion apparatus?

BIEMANN: Actually it came with a pyrolzying filament, which was sitting right 01:31:00under the ion source. The idea was to pyrolyze non-volatile compounds into the ion source, which of course ruined the compound and you had then to piece the information together to guess what it was. But we converted the Bendix insertion apparatus to make it possible to just vaporize the sample into the ion source.

GRAYSON: So you could bypass the gas inlet system altogether.

BIEMANN: We didn't really use it, because with the Time-of-Flight instrument you saw the spectrum on an oscilloscope screen and you could take a Polaroid picture of that spectrum; which worked okay below mass 150, at best 200; and it had 01:32:00relatively broad and fuzzy peaks. But once we transferred that methodology to the CEC instrument, we got good mass spectra. Then all that had some influence on the manufacturing side of mass spectrometry because now there was an area where high accuracy and long-term reproducibility of the spectra wasn't important anymore. One could scan the instruments faster and record the spectra 01:33:00faster. This made the recording of effluents from a gas chromatograph possible, but since we first used it on a Mattauch-Herzog instrument that recorded the complete spectrum simultaneously on the photographic plate, that wasn't so much of a problem. But, recording fast became important when the photographic plate was not used, so then we developed the online computer recording of GC-MS data. For that we got an IBM 1800 computer in my laboratory which made it possible to record data directly into the core memory.

GRAYSON: I'm a little bit curious about the 110, you were using it for organic applications, so you got an EI source in that instrument?

BIEMANN: Yes.

01:34:00GRAYSON: But my understanding is that originally the instrument was designed as an inorganic instrument with a spark source.

BIEMANN: A spark source, yes.

GRAYSON: Do you know what motivated CEC to develop an EI source for the instrument?

BIEMANN: That may stem from John Beynon's work at ICI Imperial Chemical Industries. John Beynon had built a Nier-Johnson double-focusing mass spectrometer for ICI. They were interested in trace analysis of organic compounds particularly in order to figure out the processes which their 01:35:00competitors used in synthesis. Dye stuff was ICI's main moneymaker. John Beynon built that mass spectrometer to look at the traces of left over starting materials and intermediates in the final product to figure out how they made it. He felt that to do that he should use high resolution so he could determine the elemental composition, because those were usually aromatic compounds with halogen atoms attached to them.

01:36:00He built that and the design was taken over by Metropolitan Vickers [Electrical Company] which was a precursor of AEI, Associated Electrical Industries, which later on became Kratos. His instrument was officially named the MS-8, which was never produced because it was not a Metropolitan Vickers product anyway. Then Metropolitan Vickers used that design and concept to build their own high-resolution mass spectrometer, which became the MS-9. It was a Nier-Johnson 01:37:00geometry that didn't have a focal plane, only a focal point. It had to use an electron multiplier to detect the ions and record the spectra. You had to scan it or you had to do what was called "peak matching", namely to put one known ion onto the detector and then switch accelerating potential to get the unknown ion into focus on the detector slit and then calculate the relative mass ratio from the accelerating potential ratio. So you had to do one ion after the other. 01:38:00Since CEC had that double focusing instrument for inorganic analysis, they could just put an EI ion source on it. They could just build an electron ionization source for that and record the entire spectrum on a photograph plate without scanning.

We then developed the means of measuring, in semiautomatic and then automatic ways, the position of all the lines on that plate, and from the position you could calculate the exact mass down to a part per million.

GRAYSON: The photographic plate had an advantage in that it was an integrating 01:39:00detector, whereas you really had a mass spectrograph with the 110 while the MS-9 was a spectrometer with a point detector. In terms of sensitivity it seems the CEC instrument would be more sensitive since you were able to integrate the signal continuously from all the ions that are produced in the ion source.

BIEMANN: You could of course, the longer you exposed it, the more signal you collected. But the only thing that was very difficult to get was the exact intensity measurements because you had to convert the blackness on the photographic plate to an abundance of the ions striking it, which is not a linear function. But again, we didn't care about whether that peak represented 01:40:0010,000 ions or 10,050 ions. All we were interested in was the exact mass of that ion and the relative abundance. In other words which ones were the abundant ions, and which ones are minor ions. That was done easily by looking at it on the microscope.

GRAYSON: You bought the 110 instrument sometime in the early 1960s.

BIEMANN: Yes. It was bought 1962, the first one.

GRAYSON: This is at the same time you wrote a book on mass spec?

BIEMANN: Yes.

GRAYSON: Were you teaching any courses?

01:41:00BIEMANN: Yes. There was a course in analytical chemistry that was in four segments, one each semester, so it was a two-year cycle, and I taught the organic analysis part of that. I had to teach one term every two years. My teaching load was always very light. One reason was the department felt my research was very important and I was doing a good job at it, also not having been brought up in the American strict course system, I wasn't too well adapted 01:42:00to going to class and teaching a lecture, then giving problems, giving exams, all the time. Things worked out well. At one point when I got the NIH facility grant which was a big grant in terms of money, people, instrumentation, and effort which I had to devote to it, I had an arrangement with the Dean of Science, who at that time was Jerry Wiesner, to not do any teaching at all, only once in a while teach a course in high resolution mass spectrometry.

01:43:00GRAYSON: Did tenure come somewhere in there?

BIEMANN: I was appointed assistant professor in 1959, associate professor in 1962, without tenure, and tenured full professor in 1963.

GRAYSON: This is a fairly long period of time after you started, about eight years.

BIEMANN: Short; from starting as an instructor in 1957 to 1963, six years. From assistant professor it was only four years.

GRAYSON: I'm sure writing the book helped.

BIEMANN: Yes.

GRAYSON: What was the inspiration for that?

BIEMANN: First, I was asked in 1960 to write a chapter on mass spectrometry in 01:44:00organic structural chemistry. I wrote that chapter and it became obvious that there was a need for a book on mass spectrometry. I had all this information, because most of it was done in my laboratory. McGraw-Hill, a publisher that had just done a book on NMR, a fledgling technique at that time, and they did 01:45:00Djerassi's optical rotatory dispersion, suggested to me after they found out about that chapter, to write a book. That was a tour de force thing of writing a book. It got published almost four years after I had run my first mass spectrum.

GRAYSON: That probably helped with your tenure position and established you pretty well in the field. I know McLafferty had done an interpretation book, was that out by then, do you recall?

BIEMANN: No, it was not.

GRAYSON: Because it's used primarily as a textbook.

BIEMANN: That was quite a bit later.

01:46:00GRAYSON: I understand that you did have problems publishing the alkaloid work because of the fact that you were determining a structure by a new method and not the old method. Were there any other details about trying to get through that publication that you left out of your literature?

BIEMANN: No, I think that was probably about it. Except that there was the question in general of how mass spectrometric data should and could be used instead of the then obligatory element analysis. I had some correspondence with 01:47:00Dr. Richard Gates who was the editor of the Journal of the American Chemical Society, so I came up with some rules or definitions as to what has to be recorded if one wants to use mass spectral data, particularly to establish a molecular weight or molecular composition. That was before high resolution mass spectrometry. That alkaloid work came a little bit later because we already used elemental compositions there.

GRAYSON: Normally papers go out to reviewers, but you're saying the reviewers were okay with what you'd done, but the editor of the Journal of the American Chemical Society objected?

01:48:00BIEMANN: Yes. An associate editor of the Journal gave me a hard time because I guess he was worried that now everybody would say "This is a mass spectrum of that compound and therefore the structure is such and such -- that's it -- believe me". Originally, he wanted us to provide melting points and combustion analyses for each one of the alkaloids, which we had found in that plant, and claimed that we actually have isolated and determined the structure of. The entire idea of using mass spectrometry was to eliminate all those steps, and all 01:49:00that wasteful burning of valuable compound just to get the carbon, hydrogen, nitrogen and oxygen values. The oxygen number was a difference anyway, so it didn't really mean much.

Eventually, he was convinced and softened it a little bit, but one of the things he indirectly complained about was that we didn't have the compound in a bottle, nicely crystalline, to take a melting point. It was one relatively valid point at that time and since it was very common that if you had a compound to 01:50:00determine the structure and thought it was identical to one which had been found before, that you wrote to that person and said "Please send me a small sample so that I can do a mixed melting point."; which at that time was the absolute criteria for identity or non-identity. Since we never had it in a bottle and there were only a few of them that we could crystallize, that wasn't possible. So he said "How would people be able to tell that they had isolated the same known compound for which you had already determined the structure because you can't send him the sample and he may not have a mass spectrometer." Few laboratories had a mass spectrometer at that time

01:51:00I said that the person could send me a very small sample, much less than was required for a melting point, and we could run the spectrum for him, and show whether it was identical or not. That finally set the associate editor's mind to rest. In addition to the fact that the reviewers who were more familiar with what we were doing. In the alkaloid community word got around very quickly about our work, because they either read my papers or I had contacted them for samples of some known compounds.

[END OF AUDIO, FILE 1.1]

01:52:00GRAYSON: We are talking about the mid-1960s and you have really pretty much established your career at MIT and now you say you actually had a second 110 instrument purchased at this time.

BIEMANN: Yes.

GRAYSON: And you had the 1800 IBM computer?

BIEMANN: We got that because of the importance of mass spectrometry in organic and biochemistry, particularly in organic chemistry and natural products chemistry and this became obvious to NIH. They thought that other chemistry 01:53:00departments would want to buy mass spectrometers, but the problem there was that nobody was trained in the use and operation of the mass spectrometer, and particularly not in the interpretation of the data. So NIH came to me and said that they wanted me to apply for a training grant, which NIH just started, a program to improve training in various biomedical sciences. I told them to explain to me what's involved and then I said yes. Then I need another high resolution mass spectrometer at that time. And I needed my own computer. Up 01:54:00until then we put all the data, particularly from the high resolution mass spectrometer, onto IBM cards, took them to the MIT computer center and had them processed and picked up the processed data. With more people working in my laboratory that wouldn't work. Of course, I wanted to have my own computer.

GRAYSON: When you say IBM cards, you mean IBM punched cards. At this point in time, the technology was such that each line of code and each piece of data required one punched card.

BIEMANN: Yes. We had automated our photoplate reader. We attached to it a 01:55:00cardpunch, which could run automatically. And the line positions and intensities were punched as maybe four or five data pairs on one card, that was as much data as could fit on a card. All that ran up the required budget quite a bit, and a training grant really was more to support graduate students and post-docs, not instrumentation. But NIH had just started yet another research grant category, the so-called "Research Resource" grant, with which we could buy all kinds of 01:56:00things. So I put in an application for a training grant and for a research resource grant which then provided a second high resolution mass spectrometer, an IBM 1800 computer, and in addition money to put the laboratory space in order, because I needed also more space.

Space is the number one problem in universities. Money is the number two problem. Everything else is below that. But it so happened that a new biology building had just been built and for some technical reasons there was an empty basement. They hadn't planned for the basement so there was no money to put any 01:57:00walls in it, and MIT didn't need it for faculty because the building itself held them. NIH paid for outfitting that part and even for the one room that was completely electromagnetically shielded for one of the two high resolution mass spectrometers. We wanted to do very long exposures for which any influence of outside electric or magnetic fields would be detrimental. It turned out that it was actually necessary because when the subway pulled out of the nearby subway 01:58:00station the current was affecting the entire area around it. We could see that on the mass spectra. It penetrated the shielded room because of the low frequency of that event. By shielding the part of the flight tube between the ion source and the electric sector, which was the longest exposed part of the flight beam, we could handle that.

GRAYSON: You picked up the electromagnetic field from the current draw when the subway went by?

BIEMANN: Yes, when it pulled out of the station.

GRAYSON: This caused the beam to--

BIEMANN: Move.

01:59:00GRAYSON: Diffuse, to move.

[RECORDER OFF]

GRAYSON: You picked up the electromagnetic field from the current draw when the subway went by?

BIEMANN: Yes, when it pulled out of the station.

GRAYSON: This caused the beam to--

BIEMANN: Move.

GRAYSON: Diffuse, to move.

BIEMANN: It moved in what's supposed to be a field-free region. It wasn't field-free for maybe 15 seconds every five or ten minutes. Anyway, this large NIH grant provided us with our own computer and the IBM 1800 model was the only one that was actually designed as a process control computer, which wasn't used as much as IBM thought it would. But it had the proper characteristics which lent itself quite well to recording streaming data, which was important for us 02:00:00to record the signal of the microdensitometer which we used to read the plates, and then later to the GC-MS online operation. That led us into lots of computer programming because we needed to write code. And of course, there was none available.

GRAYSON: You had this shop established by the mid-1960s?

BIEMANN: That grant started in 1966, and terminated in 1996. The NIH grant, the 02:01:00peptide grant as we called it, was established in 1958, and terminated in 1996. Those two were some of the longest lasting NIH grants.

GRAYSON: Yes. Was that renewed on an annual basis?

BIEMANN: At first it was three years, and then five years.

GRAYSON: What was the typical budget for those per year?

BIEMANN: The peptide grant started out with $10,000 a year, and ended up with probably $250,000 per year. The facility grant in the first year started with a 02:02:00large amount but normally ran about $500,000 a year, except in those years when -- for example we bought later on in the mid-1980s the Tandem Mass Spectrometer which itself cost over $900,000 -- in that year it was close to one and a half million dollars. MIT made quite a bit of money on my overhead, much more than it paid me salary. But that's the way it goes.

GRAYSON: It seems like you were able to get funding from NIH fairly readily, they even came to you and offered opportunities, is this a little unusual?

BIEMANN: Yes. Of course, I was really very lucky in that respect, namely that I 02:03:00started my academic career when federal research funding started to be significant. That had to do with Sputnik, when the Russians launched their first satellite. The U.S. was behind, that was a big awakening that something needs to be done and better be done quickly. At that time, money was reasonably available, World War II was long past, Korean War was past, and so there was no 02:04:00serious budget problem. They could put money into NSF and NIH. I rode up that steep slope of the federal funding curve until it leveled off and got tough. But by that time, my research program, after the late 1970s was exclusively NIH. When that leveled off it became difficult to get funding, but I was very well established, with a good track record. By the time I retired I didn't care anymore.

02:05:00When the NIH came to me, for those two programs my Research Resource grant was the first one in mass spectrometry. And Fred McLafferty got the second one a year later, when he was at Purdue.

The same thing happened with NASA. At one time in the early 1960s somebody from NASA came around. He was sort of a talent scout. It was Jerry Soffen who at that time was at JPL. He came to see what we were doing and I showed him some mass spectra of free amino acids, which we had measured on the Bendix Time-of-Flight 02:06:00instrument. And he said, "Oh, you have data!" because at that time, particularly in the space research area, people were mainly thinking and planning. That led to my involvement in the Space Program, first with the Apollo Project, and then with the Viking Project, which were sort of sidelines of my research. I called it "scientific charity", because it wasn't really something that I was interested in at the outset. But the methods I had developed lent themselves 02:07:00well to that, so if it should be done it might as well be done well and by people who are experts on it.

GRAYSON: The involvement in this Space Program was another situation where they came to you?

BIEMANN: Yes.

GRAYSON: Then you said you had run the free amino acids on the Bendix Time-of-Flight, were these small standard amino acid--

BIEMANN: Yes, a few natural amino acids to see what their mass spectra looked like, and also other things like carbohydrates and nucleosides and nucleotides. We tried to cover the biochemical area from all sides, and all those types of compounds are not volatile so putting them directly into the ion source was the 02:08:00way to do it. We translated that to the CEC instrument to get better spectra. But we never did really anything except show that one can get the mass spectra of those compounds, and what they looked like, and what one can tell from them. But it was only much later that we got into carbohydrate type things.

GRAYSON: I'm familiar with the Viking work but what happened on the Apollo Project?

BIEMANN: We checked whether there were any organic compounds, and if so, what was on the lunar surface, from the samples brought back on Apollo 11, then 12, and of course 13 didn't work, and 14. By that time, we had established that 02:09:00there were no organics on the moon, at least on the surface, and did not continue on 15, 16, and 17. But, because of the worry that there could be pathogenic microorganisms on the moon, everything was quarantined for three weeks including the astronauts. We had a mass spectrometer down at the Houston Space Center, at Johnson Space Center, where the so-called lunar receiving laboratory had been built, and half of it was completely biologically isolated. 02:10:00The astronauts lived there for the first three weeks. We had a number of instruments behind what's called the biological barrier and we had a mass spectrometer there to analyze the samples that they brought back. I was there and two or three of my graduate students and post-docs and a technician--Bob Murphy was one of my graduate students who participated in that.

We had to go through a shower to enter the laboratory, and through a shower to get out of it. We couldn't take anything with us because of this barrier, so the 02:11:00data where Xeroxed through that wall by having a Xerox machine inside but the paper came out on the outside. It was an interesting way of doing things. That was to certify that the samples are not hazardous to humans, so that they could then be distributed to other research laboratories to carry out more detailed and specific investigations on them.

After that, we got the samples into our laboratory and used a high resolution instrument and again Bob Murphy did a lot of that work. There were other problems with the lunar samples; there was worry that they could be stolen and 02:12:00sold for a lot of money. NASA shipped them to an office at MIT and the way of shipping highly classified materials was that they had to put it into a safe and in the morning we had to go get a sample, sign for the sample, weigh it, then take it to the laboratory, and at the end of the day return it there, weigh it again, and the weight difference was what we used in the mass spectrometer.

No sample was ever stolen except I think one which was then put somewhere in a mailbox, and mailed back to NASA just to show them that the security was not 02:13:00that tight. It turns out that they brought back so much material because there was no problem collecting it and bringing it back. I think about 95 percent of the material is still in the vaults at the lunar receiving laboratory.

GRAYSON: What types of analyses did you perform on these samples?

BIEMANN: We directly heated it in the ion source of the high resolution mass spectrometer and did some extracts with solvents and looked whether there was anything. But there wasn't anything.

GRAYSON: That's not terribly surprising considering the environment that the samples experienced for most of their life.

BIEMANN: But of course, nobody knew that; I remember [Harold C.] Urey, who was 02:14:00on the scientific committee and led the planning of what to do. Since one didn't know how much sample would come back, there was the question of who should get which sample, and how much. There was a competition between the organic geochemists, the inorganic geologists, and mineralogists, and at that time there was a hypothesis that the moon could be rich in meteorite material that was called carbonaceous chondrite; because they contained carbon material, and were 02:15:00chondrite type mineralogically. And he (Urey) at one point said, "I think the moon is a carbonaceous chondrite, but if it isn't the mineralogists can have it all." And it turned out not to be a carbonaceous chondrite. We had done, with NASA's support, some studies of meteorites and carbonaceous chondrites; John Hayes was one of those who worked on that.

GRAYSON: There is an interesting little anecdote that I ran across with respect 02:16:00to the Apollo mission. There is an aircraft carrier that is now a museum on the west coast probably, in the Bay Area, and when I was out there visiting my son we went and looked at it. Maybe it was the one that was used to pick up the astronauts after they had come back. I was wandering around and I saw this little plaque on the wall, which I should have taken a photograph of. It was a custom's form about the fact that these astronauts were bringing in material from outside of the country. It's an amazing kind of bureaucratic statement that they had to sign a form they're bringing back rocks from the moon, and bringing them into the country. (Laughter)

BIEMANN: But what did it say in that box where it says "Value?" (Laughter)

02:17:00GRAYSON: I don't know. But I'm going to have to go back and take a picture of that. I wish I had when I saw it. While you're continuing to do the science that you want to do, you're also doing this science for charity, so to speak.

Then you were involved also in the Mars project, was that the Viking? Were you trying to look for life or signs of organic compounds? This came naturally from your involvement...?

02:18:00BIEMANN: Involvement in Apollo. In 1968, NASA proposed and was authorized to fly a landing mission to Mars. Before that it was just a Mariner fly-by. One of the objectives was to look for any signs of biology, past or present, associated with a search for organic compounds because it was not only related to potential biology questions but also, of course, to the origin and past history and 02:19:00present state of the chemistry of Mars. Since no one had an idea what to look for and had to be prepared to find any kind of compound, I proposed to use mass spectrometry because of its wide general applicability and sensitivity; a gas chromatograph for separation and the mass spectrometer for identification. That proposal was accepted in 1969, and the rules of the game required that each 02:20:00experiment had a team of scientists to do the experiment and have the capability of interpreting the data.

That team was constituted in 1969, and I was appointed team leader. We were told that the Jet Propulsion Laboratory had a miniaturized gas chromatograph from earlier lunar proposals, but was never sent, and also a miniaturized mass 02:21:00spectrometer. So, the instrument side was all set and all we needed were people who could make sense out of the data. That turned out to be not really the case. They had built a miniaturized double-focusing Nier-Johnson mass spectrometer planned for some atmospheric studies on earth in rocket flights and it was just sitting there. Al Nier had done very successful flights in the upper atmosphere with his instrument; ironically, he used Mattauch-Herzog geometry, not a Nier-Johnson system.

02:22:00JPL's instrument had to be redesigned because the miniaturized mass spectrometer part was okay but the gas chromatograph was not suitable, nor was the data system suitable. The original plans called for doing all the interpretation of the mass spectra automatically on Mars, and send the results back. The data interpretation was to use our low resolution mass spectral identification 02:23:00algorithms. I said that wouldn't work. We had to get all the raw data back because we had by that time developed mass chromatograms, data interpretation modes which were based on having complete mass spectra of each scan, of each mass spectrum of continuously recorded spectra coming from the effluent of the gas chromatograph.

I wanted to have all the data because only that way could we be sure. The reason for not doing that was the data limitation for recording it on the lander and 02:24:00then sending it back to the orbiter, and from there sending it back to Earth. Fortunately, JPL had developed a tape recorder that didn't use a tape because that couldn't be sterilized. One problem with the entire Space Program at that time was that in order to go to extraterrestrial bodies, the rule was that you could not contaminate that body, like Mars, with terrestrial living things. Therefore, to get the Viking landers there, they had to be sterilized, and that required heat sterilization.

02:25:00So because you could not heat up and sterilize a tape, JPL had developed for some earlier un-manned lunar missions which were never flown, a recorder that didn't use a plastic tape. It used a metal wire. They could revitalize that recording system, and it was put on Viking. That also made it possible to record 02:26:00a large number of images, which required much more data than a simple GC-MS data set.

GRAYSON: They actually recorded the data on the tape recorder and sent it back?

BIEMANN: You had to wait for the orbiter, which circled around Mars on a one day Mars schedule and was in view of the lander for forty-five minutes. The data had to be stored for that time, and then quickly spewed up in those 45 minutes to the orbiter, which had a more powerful transmitter to send it back to Earth.

02:27:00GRAYSON: That made it possible to do the whole data set?

BIEMANN: That was the only way it could work reliably, because a compound that was found was perhaps not of the type that an algorithm could interpret. Even on earth such automated systems don't work for everything.

But what was missing was an appropriate gas chromatograph, gas chromatographic 02:28:00column, and the interface to the mass spectrometer. At that time the interface which we had developed, pumped off the helium carrier gas and the one which Ragnar Ryhage in Sweden had developed also had to pump off the helium. There was just no pumping capacity that could handle so much helium, particularly since helium is difficult to pump with a getter pump, an electromagnetic pump, which was the only one we had available. We finally devised an interface where hydrogen was the carrier gas that could be removed from the effluent through an 02:29:00electrolytic process through a palladium wall, that passes hydrogen very easily; but then you still have to pump on the outside.

The pumping was done by an electrical potential pump, and so we could remove 99.9 percent of the hydrogen. Because the instrument had to be pumped down and sealed off in January of 1975, then shipped from JPL to Martin-Marietta in Denver for installation into the lander and then from there to the Kennedy 02:30:00Spaceflight Center to put into the spacecraft in June. Then it had to be launched in September and spend ten months in space, land on Mars, get turned on and work. So it all worked perfectly.

One problem was that all the valves had to be tight, so the hydrogen didn't get lost. Therefore, the hydrogen was sealed off from the tank to the GC with a glass break seal that could be broken with a magnetically held piece of iron. 02:31:00Just before the launch some engineers, who had the task of figuring out all the possible failure modes that could happen, came to the conclusion that the chances that the seal was not going to break was not insignificant and they can't take that chance. They said it must be broken on earth to make sure that it was actually broken because if it wasn't, they had to take it out and put in the spare model. Therefore, the seal was broken and hydrogen was available during the interplanetary flight. We could actually run a blank after it had 02:32:00left earth and before it got to Mars. This turned out to be very fortunate because it showed a spectrum of traces of the solvents remaining from the cleaning of the instrument and it showed that the mass spectrometer worked and showed what the contaminants were. The leak rate of the valves was such that we didn't lose any hydrogen, and we established the instrument background before it ever got into the atmosphere of Mars.

GRAYSON: So what was the column like for this?

BIEMANN: The column was a micro-bore column and the material was developed by 02:33:00Milos Novotny at Indiana University. After the project was approved it was changed while under construction, it turned out that the column that was originally going to be used didn't work quite well, and we needed a Tenax column.

GRAYSON: The selection of the column seemed to be a bit of a challenge since you don't know what you were going to be analyzing . . . was there some pyrolysis involved in this?

BIEMANN: Yes, there were two. Originally, it was designed to have two modes of 02:34:00operation. One was a direct heating into the ion source; which is what we did for the meteorites and the lunar samples. The other was to send the sample through the gas chromatograph. Of course that was a period of the Vietnam War and the federal budget, including NASA's was cut. It turned out that we had to scale down not only our instrument but also others on the lander and drop off 02:35:00one of the biology experiments and delete that from the payload. We also had to reduce the cost of building the GC-MS which meant the deletion of the direct over. So we were left with heating the sample onto the GC column at various temperatures, ambient, 250 degrees, 350 degrees, and 500 degrees Celsius. We could choose various ways of operating the instrument. It also analyzed the 02:36:00atmosphere and that did not go through the GC but directly into the MS.

There was a very complicated valving system, between the GC and the hydrogen separator because we didn't know how much material there was and the ion pump or sputtering pump had to be very small and used the magnetic field of the mass spectrometer, so it was part of its flight tube. It had a very limited pumping capacity. We expected to get at least some water coming off that was absorbed on 02:37:00the material, or in part present as hydrates of minerals, and would overload the pump. This valving system operated by a feedback circuit from the ion pump current to a valving system so if at first the ion pump current became very high that meant that there was a lot of stuff coming into the mass spectrometer, it would open one valve after the other, of a gas dividing circuit, so that we could cut the flow into the MS down to one-third, one-tenth, one-thirtieth up to one-eight thousandth; and then after that, it cut it off completely. Another valve controlled the inlet system for the analysis of the atmosphere.

02:38:00Since the atmosphere was known to contain mainly carbon dioxide with some carbon monoxide, and we wanted to look at trace constituents like noble gases, we devised a chemical scrubbing system which absorbed the carbon dioxide, oxidized the carbon monoxide with silver oxide to carbon dioxide and created a vacuum--a low pressure in the sample volume so we could open the valve to the atmosphere again to get in more atmosphere; remove the CO2, and CO repeatedly so we could enrich the trace gases. That way we could measure the amount of nitrogen that 02:39:00was there, and the amount and isotope distribution of the noble gases. All of that could be commanded from earth.

GRAYSON: Isotope distribution would be interesting to the geochemists.

BIEMANN: That isotope distribution is now used to tell whether a meteorite is originally from Mars, by looking at the noble gases, occluded in the mineral particles of the meteorite.

GRAYSON: Al Nier was part of the Viking team, did you guys interact at all?

BIEMANN: Yes. In fact, he had his own experiment, looking at the atmospheric composition during entry and descent of the spacecraft. But, it could not 02:40:00survive to the surface because it was put on the part that had to be thrown off to be able to analyze the upper atmosphere. It was one of his Mattauch-Herzog instruments. He was in the middle of the project and joined my team, the organic analysis team, because some advisory group outside of NASA said that we were flying a complicated instrument, but we had nobody on our team who was an instrumentalist. They wanted Al Nier on the team so he joined our group. He was 02:41:00leading his team and was a member of my team. That led to a close friendship with him, whom I had known before that.

GRAYSON: When did the Viking Project terminate?

BIEMANN: The first lander, landed on 20 July 1976 and the second one on 3 September 1976. We didn't find any organic compounds, which some people still don't want to believe because they want to believe that there is or was life on 02:42:00Mars; and that would not jibe with no organic compounds there. We'll all just have to wait until another lander gets there that's equipped with the appropriate instrumentation or even better, until real samples come back to 02:43:00earth that can be looked at, at leisure, if such a thing exists in science.

02:44:00[END OF AUDIO, FILE 1.2]

02:45:00GRAYSON: We're in the middle of your career in the 1970s. Let's explore the business with analytical chemistry in an academic setting. Many schools treat 02:46:00analytical chemistry as a poor stepchild to chemistry. How do you feel about it?

BIEMANN: It depends on which side you view it from. As I mentioned when Art Cope was charged with making a new high power front line chemistry department, it 02:47:00also contained an analytical chemistry division. It flourished for a while. Cope was a dictator of the same class as my professor in Austria was, but of a completely different mentality. He was not a general, but rather a very benevolent dictator. That was important at that time because if you want to create a new energetic faculty out of a low level one, you can't have faculty meetings to discuss new appointments to replace old ones, etc.

02:48:00He made his own decisions, and consulted with the new faculty members he brought in because they were the ones making the place run. That kind of a faculty leader is good for that purpose, to generate something new. But when it's all running well then they run into difficulties. If a scientist starts a high tech company, but then wants to be the CEO, the CFO and the human relations person and everything else forever, it goes to pot. He's wise to turn things over to business people.

So Art Cope finally resigned under slight pressure in 1964, after almost twenty 02:49:00years as the Head of the Chemistry Department. Part of the malcontent amongst the faculty was that some didn't like analytical chemistry. Mainly the physical chemists considered the analytical chemists as failed physicists or failed physical chemists, which had some truth to it. Analytical methodology was either physics, or physical chemistry, not even organic chemistry.

02:50:00The conventional analytical chemistry of that time, in the early 1960s, fifteen years after World War II had ended, was to analyze for elements or molecules in some medium, either qualitatively -- but more quantitatively. That doesn't lend itself to great research advances that catch the public's eye or even the 02:51:00chemists' or scientists' eye.

I was in that group. But, I didn't have that problem because I wasn't pretending to be a physical chemist. I wasn't pretending to be a classical analytical chemist -- because I wasn't even trained in that. I was an organic chemist who had earned his credentials at the Institute as an organic chemist, and I was completely acceptable as a peer to the organic chemist, to the physical chemist, and the inorganic chemist because they couldn't care less. Dave Hume was mainly 02:52:00an electrochemist of inorganic background, and so was Buck Rogers to a certain extent, he was also in separations. Separation at that time just became involved with all the chromatography that was going on.

That was an important field that contributed to lots of fields including biology. But once the inventor developed a procedure, he didn't have anything to do with it -- didn't have any problems to solve with it. He had to turn it over 02:53:00to the physical chemist, to the biologist, or to the organic chemist. That created sort of the aura of service activity; either while you developed it or thereafter because you ran it for those people. Then you just turn it over and start on something new.

Now all that changed with the advent of instrumentation, starting with ultraviolet, infrared, and NMR and then eventually mass spectrometry. People took a physical phenomenon and used it to solve chemical problems. That opened 02:54:00up a new area of activity for analytical chemists and also washed out the lines. They could either turn it over to the end user, who was in the other discipline, or they became part of that other discipline. In that aspect, I am an extreme example of an "analytical chemist" who wasn't affected by that shift in views because I started out as an organic chemist; had my training in organic chemistry; but had passed through an analytical division and then established my own discipline almost.

02:55:00With the resignation of Art Cope, which was somewhat forceful, the anti-analytical forces took over and the non-tenured people were not kept. Fortunately, I had already tenure. If I hadn't, I would probably have gone back into organic chemistry, either at MIT or somewhere else.

There was only Dave Hume and Buck Rogers left. Buck Rogers once wanted to have a substantial salary increase and more space where he had to put his fist down on 02:56:00the table and say, "I want this." This was still under Art Cope at the time and Buck said, "I have an offer from Purdue University." Cope said, "Okay, if that fits you better, you're not out in the cold, it's a very good offer (it was the head of the analytical division there) so why don't you take it." Rogers went to Purdue with the task of re-invigorating the analytical division there, which is now one of the best known in the country.

After quite a few years, he left for Athens, Georgia where he stayed for the 02:57:00rest of his life. He was the one who hired Fred McLafferty because he knew from MIT firsthand what mass spectrometry could do, and he knew Fred who was at that time at Dow Research Laboratory in Framingham, right outside of Boston. So he asked Fred to come to Purdue, and Fred at first said no; but then he went. So that took care of -- I shouldn't say took care of -- the leaving of one of the two other tenured people in analytical chemistry at MIT.

Then there was Dave Hume who was more analytical inorganic. He had been working 02:58:00on the Manhattan Project and came to MIT after the War. Until the late 1950s, early 1960s MIT had a large center grant. It was a huge amount of money to establish and run the Research Laboratory for Electronics. After World War II there were lots of electronics, things had been developed, and started so MIT grabbed a big part of that support and had the laboratory where people of any department at MIT could find a funding home by working on something which fell 02:59:00under the rather broad cover of the Research Laboratory of Electronics, and get their research funded from there.

GRAYSON: Was this a part of the chemistry department?

BIEMANN: No, that was an Institute-wide thing. It resided in electrical engineering but not in the department. It was separate, like any other inter-departmental institute wide entity, and was spread over different buildings because they had to find room wherever they had. Most of this research activity was going on in the various departments, and individual faculty members who had something to do with electronics. It was rather broadly defined, so analytical methodology fell under it. Dave Hume had research support from that - 03:00:00besides a small NSF grant -- and most of his money came from there. The Laboratory of Electronics was dissolved sometime during the 1960s because funding started to become more difficult, particularly for such a grand scale -- I shouldn't say ill-defined, but--

GRAYSON: --Broadly defined.

BIEMANN: Broadly defined activities. They started to cut down, and of course, chemistry at that time didn't have that much to do with electronics so it was one of the first things that was not renewed. That's when the annual support 03:01:00disappeared. It became more difficult to get new grants for people who hadn't been supported by NSF or NIH, Department of Defense, NASA or someplace like that. Hume had troubles getting research money and his graduate students were working more in civil engineering on water pollution problems. His research declined for various reasons, and MIT had an early retirement program where as soon as you got in the sixty to sixty-five age bracket you could retire. At that time it was mandatory to retire at age 65. So, he decided to take early 03:02:00retirement because it was most beneficial and that was the end of him.

On paper there was an analytical division of which I was the only faculty member. But for practical purposes, it didn't exist anymore. I ended up having all the analytical graduate students. All the lectures in analytical chemistry I had to give, but only in two-year cycles. All the exams, like the general exams, and the various exams the graduate students took, I had to write. I knew exactly what each of those students knew and didn't know. It was very hard to write a 03:03:00three-hour written exam for the class, which was to apply to all of my graduate students.

I finally told the department head at the time that this arrangement didn't make sense anymore. All of my graduate students always came to MIT because they wanted to work in my research group; it was not that I interviewed incoming graduate students and all of them talked to me, and then one made a decision. That was just due to the peculiarity of my research laboratory and its relative uniqueness. Also, whether their undergraduate mentor knew about me and what I 03:04:00was doing. Many of my students came from certain university groups--I had quite a few students from Bowdoin College because one of George Buchi's post-docs joined the faculty there in organic chemistry and so he knew from the beginning what I was doing, and he told them "Go to MIT, work with Klaus Biemann, that's a new field."

That was the end of the analytical division at MIT except the process was slowly spread over a period of fifteen years. Since I came out of the organic group, I was always in communication with everybody in the department because not only 03:05:00organic chemists were interested in mass spectrometry but also biochemists, some physical chemists, and inorganic chemists as well. So I had no problems with communicating within the department, and didn't at all feel isolated. But that's kind of an unusual position.

To get back to analytical chemistry: as I said earlier, I think it's now the instrumentation, from optical to electron spectroscopy, x-ray etc. which provide 03:06:00a wide area of research possibilities. On these you can work in any part of the department. For example, John Waugh in our physical chemistry division was very much in the forefront of the early development of NMR [nuclear magnetic resonance].

GRAYSON: Was he at MIT then?

BIEMANN: Yes. And he still is.

GRAYSON: Okay. He was part of the group, a mainliner but he was actually doing 03:07:00analytical instrumentation work.

BIEMANN: He didn't at all consider himself to have anything to do with analytical chemistry. He was doing research in nuclear magnetic resonance, both in the theory of it, and the instrumentation, building new instruments, or buying new instruments, and modifying them.

GRAYSON: But this is exactly what you were doing in mass spectrometry?

BIEMANN: Yes, I think that's what analytical chemistry has become. A lot of separation science methodology and technique was developed in organic chemistry and in biochemistry, and biology, because you need it. Therefore, you think of 03:08:00ways to do it or ways to improve what has been developed by somebody else. The question really is whether what's called analytical chemistry is needed or viable as its own little box. If you stay in the box it is kind of only making things and sending them out of the box. And IF you go out of the box, then you are not in the box anymore!! Purdue is a perfect example. Graham Cooks did an 03:09:00excellent job in all the ion traps. But, he could have done that just as well in a physics department or in a chemistry department with physical chemists or in a chemistry department, as an organic chemist. The only difference between him and me is that he was more interested in the instrumentation and the technology, and to advance that; while I was more interested in the instrumentation and the technology because I wanted to do something with it. In my laboratory we never built something or made something or invented something if we didn't need it.

For example, the GC-MS was not developed because I wanted to connect a gas 03:10:00chromatograph with a mass spectrometer. I wanted to do it because we had to collect tediously peak by peak in one melting point capillary after the other, put it aside, label it, and make sure we didn't confuse it. Then we put them into the mass spectrometer, one after another. This takes a long time. But it also gives you a lot of time to think about how to convert that tedious manual process into an automatic one, which finally led to Jack Watson's development of the fritted tube connector.

Those are just two different sides of that part of instrumentation. Others are 03:11:00intrigued by the physical principle and want to make it useful for something where they think there is a need for it. Most of the time they are right, that there is a need for it; or perhaps one doesn't hear about the ones that didn't turn out to be of any use. Therefore, there are some universities who have very well functioning and productive analytical divisions but one wonders whether the existence of that division as a management organization is really necessary to produce that effect; because the advantage of doing it in the other field is 03:12:00that you are much closer to the problems that cry out for a solution or they may be in your own laboratory. They don't have to be found out by chance, like when I went to the meeting in Chicago. They don't sit around unused because it takes a while for people to find out what you did. If you're not actually developing it for a certain purpose you publish it in an instrument journal. You don't publish it in a biology journal or in an organic chemistry journal. So the chances that things are recognized by the beneficiaries of the technique are 03:13:00lower and less quick, than if that development goes on in an environment for which it is actually predestined.

I think that's what one has to think of when one talks about "analytical chemistry" and division of analytical chemistry. As I said, some like the one at Purdue is an excellent one. The one at Northeastern is a very good one. And those are the only two ones, I know of off hand because one is very close and some of my students are there. The other one is well known in the field. Most other universities can do without the specific field labeled as such, without any disadvantage.

03:14:00GRAYSON: The lab that you started at MIT was the first laboratory that taught mass spectrometry, even in the world.

BIEMANN: You mean as far as--

GRAYSON: In terms of students coming to learn the technique and then to leave your institution and have a career based on their ability to use a mass spectrometer effectively.

BIEMANN: I think it was the first one because there was no university in the United States that even had a suitable mass spectrometer -- there were lots of isotope ratio mass spectrometry in biology because of 15N studies. There was one 03:15:00of these at MIT in biology. They had a mass spectrometer but it was a Nier-type isotope ratio machine that could only measure two masses, but very accurately. But of the organic analytic type, my laboratory was the first one at the university, in the United States. There was one group in Australia, but that was a government laboratory.

GRAYSON: Was there anything going on at the Karolinska Institute?

BIEMANN: Yes. The mass spectrometer at the Karolinska Institute was built by Ragnar Ryhage with his own hands. Professor Einar Stenhagen was at the 03:16:00University of Gotheborg. But there were no students involved, certainly not at Karolinska because it was a pure research institute. Stenhagen was a biologist more or less, and the reason why he was in mass spectrometry was because he wanted to study the fatty acids produced by the tubercule bacillus, in part because his wife had tuberculosis. As far as actually training students--of course that was one of my problems that I had to start out with post-docs 03:17:00because the average graduate student walking into MIT wouldn't have tried to work with me. I needed more well-trained organic chemists, which I fortunately had access to at the University of Innsbruck.

GRAYSON: Most of the training that is going on today in the field of mass spectrometry is spread around the country; Burlingame has an operation on the west coast; McLafferty was at Purdue--

BIEMANN: And then at Cornell--

GRAYSON: Yes, and Cornell. He (McLafferty) wasn't at Purdue very long was he?

03:18:00BIEMANN: No. All of my early graduate students went into academia and started their own laboratory and trained a lot of students.

GRAYSON: Today there is a reasonable number of places.

BIEMANN: Today's mass spectrometry is so different from what it was fifty years ago, thirty years ago, or even twenty years ago. Now it's so highly automated to 03:19:00the point that when you put in the sample and click the 'on' button it tells you what it is by searching the NIST library right away. Then even in protein chemistry what's now called proteomics, you don't have to interpret anything anymore because it's all automated with the human genome and many other genes of many other classes of organisms known.

You can, as you know, digest the protein with trypsin and separate it on a gel. Then run it either by ESI [electrospray ionization] or MALDI [matrix assisted 03:20:00laser desorption/ionization] get all the molecular weights or at least two-thirds of the molecular weights, and then it's purely a computer problem to match those with all the proteins which the genome information contains, and you'll find out what it is. And then all you need to find out is how it was modified. But again, just look at the shifts in molecular weights to see whether there is a phosphate group or not, and if so, whether it's one or two, and things like that. It's not that you look at the data and have to use your experience to interpret it. It's all very much faster. It produces so much more 03:21:00data that the human mind could not possibly do it in the intellectual way. You have to use computers to help, and it's certainly a great help. It solves the problems and you don't have to have five years experience in basic mass spectrometry.

Most of the people nowadays push the 'on' button, and don't know what's behind the panel. If it doesn't work, you call in the service man or push another button, and it tells you to check this or check that or tells you that you don't have accelerating voltage and you better check that connection. It has changed very much. Now, I think the largest use is routine applications of the method. 03:22:00It's like milk where the cream is on the top. There is a small layer of people who do actual research in mass spectrometry, and it would be an interesting paper study to see in what kind of institutions and departments that layer is. I think that there would be relatively few of them found in an analytical division at a university.

GRAYSON: There was a period when environmental applications dominated the field, at least in terms of the applications I was thinking about. Did you ever do much in the way of environmental mass spectrometry?

03:23:00BIEMANN: No. Maybe my greatest contribution to environmental mass spectrometry is the connection of the gas chromatograph to the mass spectrometer and to a certain extent the use of high-resolution mass spectrometry. Now in environmental analysis that is mainly used for halogenated compounds, particularly dioxin. That has probably declined in the last twenty years by elimination of the product which was an impurity in 2,4,5-T, the Agent Orange type herbicide. It's kind of unfortunate, because it's a great herbicide. We did 03:24:00the first experiments to look for dioxin in collaboration with Professor Matthew [S.] Meselson, a biochemist at Harvard University. He got involved in the Agent Orange situation and asked me to be part of a group looking into how one could come up with an analytical technique.

We ran some spectra on the 21-110 high resolution mass spectrometer to show that 03:25:00one can obtain a clean signal because of the many chlorines, so different in accurate mass that they can be detected in the presence of lots of other stuff with the same nominal mass. Meselson then had one of his graduate students work out the methodology on the MS-9 which they had at Harvard. Otherwise, I have not done much in environmental chemistry -- except I started Ron Hites in that field. It was his own idea to use mass spectrometry for that purpose, and he went out to collect gunk from the bottom of the Charles River in front of MIT, 03:26:00lowering a bottle down to the bottom of the river from the Harvard Bridge that goes by MIT, and collected a sample and looked at that.

Later on, I was a little bit involved when Ron, then a faculty member in chemical engineering at MIT, had a program that collaborated with other people looking at the combustion products of fossil fuels, mainly looking for carcinogenic polycyclic aromatics. I took that program over from him when he left MIT for Indiana University, for a few years until it ran out. But, it was 03:27:00never close to what I was doing, it was not biological enough to be of lasting interest. It was more of an analytical technique where pushing detection level and accuracy was very important. Then it became so politically enmeshed and legally enmeshed, and I always stayed out of things where I had to appear in court and testify for one side or another.

GRAYSON: Did you ever have to go to court?

BIEMANN: No. Once I came close which is an interesting anecdote. There was a racehorse which belonged to the Aga Khan and it won one of the New York State 03:28:00big horse races. They always tested the horses afterwards and the sample was sent to the laboratory at Cornell.

GRAYSON: Is that where Hunt was? Don Hunt.

BIEMANN: No, no, Jack Henion was there. He did a lot of GC/MS, and LC/MS and was in on the early parts of electrospray which they called a different name. He found one of the doping compounds in the urine sample from the Aga Kahn's horse. 03:29:00Not the Aga Khan himself, but his managers said, "No, we didn't dope the horse." So they sent the sample to another laboratory in the Chicago area, and they didn't find anything. A group was convened to sort things out, and I was the mass spectrometry person on it. It turned out that the evidence at that Cornell laboratory was not kept properly. The laboratory was located at Cornell University but it was actually operated for the government of the State of New 03:30:00York by the Department of Veterinary medicine and Jack Henion did the GC-MS analyses.

Unfortunately since the scientist was in academia, it was a side job for him to do those analyses and he didn't do all the blanks necessary to make sense out of it, and did not do those things on which so much rides. You have to have a very detailed flow of evidence and custody, and sign out, when you take the sample out of the refrigerator, and then sign it back in again. Those records were not 03:31:00kept properly so you couldn't even tell if the spectrum was from that sample or from another horse, or another blank or whatever. The state office which had filed the complaint finally gave up, so I didn't have to go to court over it. But in general, I tried to avoid those things since plainly that was the thing to do.

GRAYSON: That has become a fairly important use of mass spectrometry today with the animal and human sports, testing, as we found out with this bicycle race in France.

03:32:00BIEMANN: And of course the Olympics.

GRAYSON: Yes. This is a very necessary tool, small amounts of material can provide a lot of information quickly, and it's important.

BIEMANN: That reminds me of another project which was of interest in the early part of the 1960s. We developed a procedure for looking for drugs and 03:33:00metabolites in newborn babies and children, mainly the accidental ingestion of things which they found in the bathroom cabinet; overdoses, un-intentioned, rarely any foul play was involved. But it was important to identify it quickly and reliably. That was just after we had developed GC/MS and used it for alkaloids for example. We made a connection with an anesthesiologist at Harvard Medical School and then developed that as sort of emergency service.

03:34:00We generated a computer program that would look through the GC traces and the mass spectra to identify what didn't belong. We had a 24 hour operation, that if a child was brought into the emergency with some funny symptoms which indicated some toxic problem, they would take the blood and the urine sample, call a cab to send it over to our laboratory and call someone from my laboratory at home 03:35:00and that person then went to the lab. The cab-driver was told exactly where to go. That was at a time when not all the buildings were completely locked and he was told to go to the basement of building fifty-six and hand over the samples. The person on call would quickly extract the two samples, run the GC-MS, figure out what it was and call the physician back and tell them what it was. We did that for two or three years, until it became established. It became so common that analytical laboratories, at least in the greater Boston metropolitan areas, made it a business to run all kinds of analyses; including having mass spectrometrists so that it could be turned over to those commercial 03:36:00laboratories. There was a need for it, with clients that would use it, so they could set up the procedure, and assign or hire someone to do it.

GRAYSON: One of the areas that I discovered where mass spectrometry is used to detect inborn errors of metabolism in newborns. We had a speaker from the state government laboratory in Columbia, Missouri talk to our local discussion group about it a year or two ago. I was really impressed with the way they go about 03:37:00it. I mean they're really not mass spec people. They're more clinicians, but the use of the tool to detect these inborn errors in metabolism is such a life-saving thing and it just gives people information immediately if there's any of these really quirky diseases that even though the baby seems normal at birth, after a short period of time their health may start to deteriorate and it's just a fascinating and wonderful application of the tool.

BIEMANN: One of the pioneers of this field was Isamu Matsumoto in Japan, at Kanazawa Medical University, who really established his entire laboratory that 03:38:00was geared at finding all those errors of metabolism by identifying the products that accumulated because some enzyme was missing. He built a library of mass spectra for that and actually had a nationwide laboratory. That was in the mid-1970s. He died ten years ago. He was a big person in medical mass spectrometry. It then spread all over Japan to different laboratories doing it. It became--not commercialized -- but with the methodology being done by certain 03:39:00central laboratories, like the one you mentioned. But, one can say it was another outcome of our original introduction of mass spectrometry to organic chemistry.

GRAYSON: You talked about moving from the accurate mass determination on the 110 to the use of tandem mass spectrometry. What kind of instrument did you have for that application?

BIEMANN: That was the JEOL HX-110. Tandem mass spectrometry has two forks: one 03:40:00is quadrupoles; the triple quadrupole, and now with ion traps and things like that.

The other one was the magnetic instruments which started with two sector instruments where one could use the linked scan mode. It worked quite well but the resolution and sensitivity was quite low. Then one went to three-sector instruments where you used the magnet as a single focusing, first mass spectrometer, and then a double-focusing geometry with an electric and a magnetic sector as a second analyzer; or the reverse. Then the ultimate was a 03:41:00four sector instrument with two high resolution mass spectrometers in tandem and in between the collision chamber.

The first of those were built by VG and went to the National Institute of Environmental Health Laboratory in Research Triangle, North Carolina. It worked reasonably well, but not good. It was two ZAB-SEs. The first one had the 03:42:00magnetic sector followed by the electric sector, but then the second mass spectrometer, for some reason or another, they turned it around so that it had the electric sector first and the magnet second. If the ion optics of the Nier-Johnson geometry would be absolute correct and symmetrical, it should work. But it's not quite perfect and therefore for some ion optical reasons it never 03:43:00worked very well.

So when I wanted a high resolution tandem mass spectrometer, I looked at that instrument but I knew that JEOL also had a high resolution, double-focusing mass spectrometer based on the Nier-Johnson geometry. I happened to be a consultant with JEOL; just starting at that time. They showed me the prototype. So I said, "By the way, you should also think of making it a tandem mass spectrometer" by 03:44:00putting two of them together. But neither of us could understand why there was any reason to turn the second one around, you might as well use it as it is designed and shoot the second ion beam into the second mass spectrometer in the right way.

GRAYSON: Electrostatic and then magnetic?

BIEMANN: Yes. I asked them to build one (tandem mass spectrometer) for me. They first took a step back because they were just getting the single one off the ground, and trying to market it. But to make a double one?? Finally they agreed to do it. But they didn't know how much to charge for it because it was the first one, there was a lot of research to do, so we agreed that they would build 03:45:00it and sell it to me for the same price as the VG tandem mass spectrometer that was on the market. I thought that was fair, so it cost them a lot of money, but I also said I need it in nine months. I explained to them that in nine months my grant year runs out and I had to spend the grant money for that instrument in that grant year. I had money in the grant for the VG instrument and that's the reason I also had that price in mind for the JEOL instrument.

If it wasn't in that year (I think it was 1985), I couldn't pay for it, and of 03:46:00course, they didn't want that to happen. They put together a completely separate group of ion optics people, engineers, manufacturing people, and management people for that project. My grant year ran from 1 August to 31 July and they delivered it on 29 July. They built the entire thing from scratch having just designed a single unit and did not sell any one of those yet, and they pulled it off. They had to do it because the president of the company, Dr. Ito, had talked 03:47:00to his managers, and the top manager said that they could do it, so he promised that to the president. From that point on, in the Japanese system, that had to happen come hell or high water.

They put everything into it and built a marvelous instrument that worked very well from the first day on and I never had any problems. They probably sold between fifteen and twenty instruments having the price increase in the end to about $1.3 million dollars, so they did make some money back. But what they spent on the first one, who knows? Of course, for them it was also a prestige thing to have it delivered to MIT and to my laboratory. So it was kind of a 03:48:00'perfect storm' in a positive way; everything came together. Then we put another level of sophistication, performance, and results on to the peptide sequencing because we used a FAB source, and could sequence a peptide, twenty amino acids long, in a single mass spectrum.

GRAYSON: This instrument worked with the FAB source, the first MS (of the tandem instrument) probably had a resolving power on the order of a couple thousand for the purpose of--

BIEMANN: Up to ten thousand, but we set the resolution for whatever was needed 03:49:00to resolve the isotopic multiplet at that mass. That gives a mono-isotopic parent ion species which we used -- although for large molecules it is no longer the biggest signal -- but it gave a simple secondary spectrum. That was the big difference compared with the triple quadrupole which Don Hunt used very successfully; but he had to use the isotopic cluster, therefore the daughter ion spectrum was polyisotopic.

GRAYSON: Much more complicated. The second mass spec (of the tandem instrument) operated at similar resolving power?

03:50:00BIEMANN: Again, depending on what the mass range was. But you couldn't run it below 1,000 resolving power anyway. So, we ran it at about 2,000.

GRAYSON: Then you could ionize the material in the FAB source and got the ion that represented the peptide.

BIEMANN: The molecular ion in very good yield with no fragmentation. Of course fragmentation was irrelevant.

GRAYSON: Right.

BIEMANN: We could do mixtures of ten peptides in one sample, no problem. We did an enzymatic digest of the protein and ran it through an HPLC [high performance liquid chromatograph]. We didn't collect single peaks but groups of peaks, so we 03:51:00ended up with the primary spectrum, maybe ten signals for ten different molecular ions. Then we could just pick off one after the other. We had to reload the sample but that was easy.

GRAYSON: So now you're using a mass spectrometer as opposed to a spectrograph?

BIEMANN: Yes.

GRAYSON: Electrical single point detection.

BIEMANN: As mentioned earlier, since the 1960s we had two CEC 21-110B instruments. In the late 1970s we bought an Atlas MAT 731 mass spectrometer that is not quite the same geometry but a more advanced model, mainly designed for organic compounds.

03:52:00GRAYSON: Was the 731 Mattauch-Herzog geometry?

BIEMANN: Yes. The 731 was Mattauch-Herzog geometry. [The 711 was also Mattauch-Herzog geometry, but not as well corrected to get a focal plane; it had a focal point. So one used it as a scanning instrument.] The 731 was the one we right away converted to FAB once we read Michael Barber's paper and it became known that the secret to FAB was to add glycerol to the sample. So that instrument remained mainly in that mode until we got the JEOL instrument in 03:53:001985. After that we didn't use photo plates anymore.

GRAYSON: Was there an electrical detection system on the 711?

BIEMANN: Yes, with an electron multiplier.

GRAYSON: At one time, weren't you using field ionization/field desorption as an ionization method?

BIEMANN: Yes. The MAT-731 had a field desorption source which we used briefly, 03:54:00but again, a year or two later FAB came about and that was just so superior that we used the field desorption source and the vacuum lock that carried the emitter wire, to convert that emitter wire stage to the stage for FAB. Then all we had to do was put on an argon gun from the top where we had a viewing port anyway, and that was all done. FAB-MS really opened up the field of peptide mass spectrometry to anybody who had a good mass spectrometer. Before that, there was 03:55:00only our chemical method, using GC/MS for it and small peptides and the permethylation method which Dudley Williams and Howard Morris developed in England. Morris came to Don Hunt's laboratory to show him how to do that chemistry because it was even trickier than our reduction method.

The other disadvantage was that the permethylated peptides were not as volatile as polyamino alcohols, which we used so you couldn't use gas chromatography. You had to fractionate or sublime the sample mixture into the ion source and scan 03:56:00continuously and see which peaks came up and which ones disappeared to sort it out. That was quite tedious. Then Don Hunt used LC/MS to do that, but again, since FAB came out, everybody scrapped everything and did FAB -- without any chemistry.

GRAYSON: Yes.

BIEMANN: Since the chemistry was the thing, you had to be very experienced to use it. This prevented the spread of the methodology; but with FAB anybody could do it. And that was really when peptide and protein mass spectrometry took off. 03:57:00But by that time the biochemistry community had already been introduced to mass spectrometry by our work; because there were problems in protein chemistry which Edman degradation could not solve. Those people then had most of it done but then needed to deal with those other problems; like when the N-terminus of the protein was blocked, which is the case in 30 percent of mammalian proteins. These post translational modifications the Edman degradation couldn't do, so then they sent us the N-terminal peptide and we determined that sequence. And that was done.

Or if the peptide was too small and it wouldn't stick in the spinning cup which 03:58:00was the heart of the Edman automatic method then that was duck soup for us. Same thing for hydrophobic peptides like those containing leucine, isoleucine or proline, which if there are too many of them left at the end of the chopped-off peptide makes it very hydrophobic and is then washes out by the solvent. But of course, the more hydrophobic they are, that means the less polar they are, the easier it was for mass spectrometry. So it was a perfect match to do those pieces by our technique.

03:59:00By that time biochemists already knew about that one can do certain things in peptide chemistry by mass spectrometry so they were ready to get going and use it on a larger scale, which FAB made possible. But even before that, in 1977 Maxam and Gilbert at Harvard developed their DNA sequencing and Sanger in England developed his, so DNA sequencing just came about as a way of indirectly establishing protein sequences by translation. But there were lots of experimental problems which we can't go into in one afternoon.

04:00:00It so happened, that Paul Schimmel in our biology department at MIT, was interested in a very large protein, perhaps 1,000 amino acids long. It had not just one enzymatic function but more than one, which had to work in concert to recognize the transfer RNA to pick up a specific amino acid and then transfers it to the growing protein chain. That enzyme had to wrap the entire thing around in proper functionalities and sequences to do that, and hand it more or less from one step to the next. These were large proteins. To do that by the Edman 04:01:00degradation, it would have taken a long time, very tedious.

Paul figured out that if he does a DNA sequence he can translate it, so we talked about it and about the problems with DNA sequencing at that time, in concept and practice. I realized that all those problems could be easily solved by our GC/MS method. So we collaborated on that; he was doing DNA sequencing and we were doing mass spectrometric partial sequencing only of the protein. We could tell where there was a mistake in the DNA sequence because a peptide didn't fit anywhere to the translated amino acid sequence. Within one year we 04:02:00finished the work on a protein that was 875 amino acids long. That area of protein chemistry got people perking up and even before that was finished someone else asked to collaborate on a similar protein. But then, even while we worked on that second one, FAB came around and so we just completely switched to FAB. Then we could do big peptides and cover much more of the DNA sequence.

From then on lots of people could start using mass spectrometry for protein work. Of course, we had spent twenty years on that GC-MS method from the first 04:03:00experiment in 1958 to the first real protein that we determined the sequence of just with mass spectrometry in 1976. So over night that methodology became obsolete. Fortunately we were in the position of working on other problems that were waiting to be done right away, and makes things faster.

GRAYSON: Looking back you can see that the real limitation with mass spec was that it could be considered to be a little bit of a niche technique in the analytical scheme because it depended so much on getting the molecule into the gas phase and then doing ionization with electron impact, electron ionization. 04:04:00It wasn't until FAB came along that it just opened up the whole range of molecules that could be studied by mass spectrometry by orders of magnitude. It became a much more universal technique with that simple movement from being free of getting molecules into the gas phase before you could ionize them.

BIEMANN: FAB also was more applicable to polar, particularly basic molecules, and not so much for the fatty acids and fats. But it so happens that biologically significant and important molecules are polar. Many of them have 04:05:00amino groups, so FAB made it even more useful for that important area, namely biology, than it would have been in any other, vaporizing technique. Of course, electrospray and MALDI [matrix-assisted laser desorption/ionization] were in the same direction; even more applicable to those things. To do fatty things by MALDI is almost hopeless, and very difficult by electrospray. But fortunately, nobody but the food industry was interested in fats. Since you can't even easily distinguish trans from cis fatty acids, that makes it even less significant for 04:06:00the cooks nowadays.

GRAYSON: Was the tandem instrument the last big purchase for your lab or did you acquire other equipment following it?

BIEMANN: No. The curious thing is that by the time of my retirement in 1996 it was hard to even give it away, because of electrospray and MALDI--well; electrospray was difficult to do on a magnetic instrument. We tried it but because it used 10 kilovolt accelerating voltage the electrical insulation 04:07:00problems were substantial in using electrospray for that kind of instrument. MALDI was also not appropriate because that's a pulse technique and the Time-of-Flight instrument was much more suitable for that. Now the Time-of-Flight instrument had, at the beginning, the problem of low resolving power. Now that I think about it, I should say that we did buy another instrument, namely a Time-of-Flight, MALDI instrument which was the first one 04:08:00that Marvin Vestal built. He knew that I had a renewal application for my research resource grant with NIH and he was a member of the site visit committee, so he knew that I had the money to buy such an instrument.

It took me quite some time to decide which one to buy because the one which Franz Hillenkamp used was really a commercially available instrument for inorganic analysis which he modified. Then that company wanted to make a 04:09:00commercial instrument for organic and biochemical applications. Marvin Vestal had been mainly working on quadrupoles for electrospray. Since he was one of the inventors of chemical ionization, electrospray was a little bit in his line. He had left the University of Utah, and started his own company in Houston, after a short intermediate stay at the University of Houston.

04:10:00At the same time, or shortly before, Brian Chait at Rockefeller University had built his own instrument, actually put a MALDI ion source into Time-of-Flight instrument which he had. So, he had more or less the design of a MALDI Time-of-Flight instrument, which he had built for his own laboratory, and Marvin Vestal made a deal with him to use his design to make it commercially available. So knowing that I have the money, he called me up and said. "By the way did you buy your Time-of-Flight instrument yet?" I said, "No, we are still thinking." He said, "Why don't I build one for you?" Since Chait had proven that it works and 04:11:00I knew that Marvin Vestal could build instruments. I said, "Okay." We ordered it, he built it and in a few months we got that instrument. Later on Perceptive Biosystems in Framingham started to build some and we bought one of those.

GRAYSON: So the Vestal instrument, that was just a standard linear Time-of-Flight.

BIEMANN: Standard linear Time-of-Flight. As far as my laboratory goes there were a number of problems. One is that I had to think about retiring, and I thought 04:12:00that at age 70 I would retire. But one or two years before that MIT decided to completely redo what used to be called the new biology building, which was thirty-five years old. I was in the basement of the building. At first, they said they would only redo the upstairs. But then, as a design and an engineering company was hired and architects plans were drawn up, they figured out that they 04:13:00couldn't completely gut and redo five stories above ground and not disturb things in the basement below. So, eventually they decided I had to move out.

But they realized that there was this huge, heavy big expensive instrument in the basement and they promised to keep it running so as not to disturb my research. But, everything else was to move out. So, it was finally decided that we would move on the fifth floor of the new chemistry building which by that time was twenty-five years old. I was already reducing the number of people in my laboratory because of my impending retirement. So that Time-of-Flight 04:14:00instrument -- actually by that time two Time-of-Flight instruments -- were moved up to the fifth floor. Of course, the big instrument couldn't go on the fifth floor. We moved up there, but then, in one of the boondoggles of MIT, they found out that they had to shut off the air conditioning to gut the building; but this instrument needed air conditioning.

They built a separate air conditioning system for that room so we could run it. But, then it turned out that we would have to wear a hard hat to get in to the 04:15:00room. This escalated to the point that I said, "Let's just put it to sleep, turn it off, and when the construction is finished we'll turn it on again." Everything was sealed off so that no dust got in there. You can imagine dust on a 10 kilovolt instrument. That went on and in the meantime fortunately the MALDI Time-of-Flight methodology improved at a relatively fast pace so that it then got to the point that one could do the delayed extraction and get sort of tandem mass spectra out of it even without having a reflectron, just a linear one. That 04:16:00we could get working, for our purposes of peptide sequencing, pretty close to the data that was more or less equivalent to what we got on the huge four-sector instrument.

While we had to shut down the instrument, it had no impact on my already reduced research activities. Then when they finally were finished it turned out that everything was covered with dust because in order to keep dust out there had to 04:17:00be a hood running in the room adjacent to the instrument itself. Because it used a laser, which had to be filled with fluorine, we had to have a tank of fluorine. However, the safety regulations didn't allow that, even though we only used it once in a while to fill up the laser cavity. If anything leaked, it had to be automatically vented, so it had to be in the hood. The hood had to run next to the tandem mass spectrometer. At one time, the people working on the reconstruction had to turn off the hood fans on the roof of the building.

04:18:00At first, they didn't know that there was one that they shouldn't shut off, so they did shut it off. But when somebody pointed out to them that that one has to be running, they turned it back on but in the wrong flow. So instead of having it under over-pressure inside -- which you don't use a hood normally that way, but in this case it had to be over-pressure -- they turned it on the wrong way. It sucked air in from the outside dusty environment. So it ran like that for a few months. Even though the room was sealed pretty well with duct tape, still dust got in. I said, "Let's not even turn it on, because I'm going to retire in 04:19:00a year or less." In the meantime others developed another way, with a much simpler instrument of getting the same kind of data.

For ten years it had served its purpose; let's just not turn it on again. Then we had to look for someone who would use, not buy it, but at least pay for the shipping, which is no simple task for a ten-ton piece of equipment. We finally found a laboratory in Boulder, Colorado, which is a government research laboratory. They only needed one of the two mass spectrometers. They wanted to use it to develop new detectors, so they needed something that generates an ion 04:20:00beam that can go up to eight, ten kilovolts of energy. They took it and I must say that I have never heard of it again. And the same thing happened with others; Catherine Fenselau had exactly the same instrument. It was just like FAB on peptide projects wiped out the need for doing the chemistry and GC-MS; so did MALDI Time-of-Flight, particularly with the reflectron on it, replace the large tandem instruments -- at least for polar molecules that one would use a tandem instrument to analyze.

Within those ten years magnetic tandem mass spectrometry did a great thing, and 04:21:00suddenly it was obsolete. That is one of the things that make chemistry and instrumentation nowadays so rapidly changing because it gets so highly developed; and in the process so expensive. People think of doing it better, cheaper and faster, and the moment that succeeds; that of course makes previous expensive, complex, very sophisticated, and very elegant instrumentation overnight useless. It is left for some very narrow fringe things in which you 04:22:00can do some detailed studies.

GRAYSON: The primary use nowadays for those old sector tandem instruments is if you want to do high-energy collision work. Most of these other instruments the collision cells operate at relatively low energies. That's adequate for most applications. But if you have an interest in high-energy collision then that's where these instruments can still be useful but not a whole lot of people do that kind of work. You retired in 1996 from MIT? You slowly wound down your operation?

BIEMANN: Yes, I wound down the operations over a period of three or four years. We closed down the laboratory in 1999. When my last graduate student graduated 04:23:00in May of 1998, I had two post-docs still for another year and then that was it. I got out of chemistry because the work I was doing, and the way I was doing it and the way it had to be done, you couldn't do it with one or two post-docs, which was the way many of my colleagues at MIT or other places operate. They 04:24:00officially retire because they don't want to teach anymore -- which I didn't have to do much anyway -- but still they can't let go. They continue to work, scrape up some money to pay one or two post-docs -- and it's even harder for those to get research grants because everybody says, "Why don't we give it to a young active person, other than to this old person who has gotten his Nobel Prize already?"

I am referring to one of my colleagues, Gobind Khorana, who had a great research career, a great research laboratory. He got the Nobel Prize for synthesizing the 04:25:00first gene in the 1960s. But he still writes research grant applications to carry on with one or two post-docs doing some nice biological experiments; which you can do with even one person to work on a project. He has fun with it. But I don't know as of today if he still does it, but at least for many years after his retirement he did that. You can do that if you study some biological process where you just need a few test tubes, a spectrophotometer, or an HPLC. But in my business without instruments, you can't do anything. To keep those instruments, even Time-of-Flight instruments, running you need a number of people to make it 04:26:00worthwhile. I could do some work by borrowing time on some mass spectrometer in the biology department; but it's not my cup of tea. As you know, just keeping up with the literature these days is a full time job.

GRAYSON: Time to hang it up. I'd like to explore a little bit more the importance of the mass spec meetings; the ASTM E-14 meetings and the ASMS [American Society of Mass Spectrometry] meetings. ASTM E-14 started very close to when you started mass spec back in 1954.

04:27:00BIEMANN: I went to the first one in 1958, when it was in New Orleans because the instrument was delivered in beginning of May of 1958. I went to the first one when I had a mass spectrometer.

GRAYSON: They were more like Gordon Conferences I would think at the time they were kind of small.

BIEMANN: Small, very informal, and just one speaker at a time. It grew out of CEC users' meetings. It started by chatting about common problems and solutions, probably not even with a real program. But, in 1958, it had a program for that--

04:28:00GRAYSON: I have copies of all of those old programs, courtesy of Charlie Judson. It's interesting to go back and look at some of the titles. There was a lot of work done that you could put in the title of today's meetings, which show how things have changed -- but they're the same. The meeting has been, I think, a fairly dynamic event for mass spectrometry. What is your feeling about the conferences? Do you feel that they have contributed?

BIEMANN: You mean today?

GRAYSON: Well just to the evolution of the technique from where it was in the 1950s to where it is today.

BIEMANN: Of course, in the beginning it was a small group of people -- I would 04:29:00think that probably 80 percent of the attendees knew each other very well. To a certain extent it was almost a social get together to chat with old friends. And of course, I was a newcomer. Always at the time, first I should say that I think it is a very good thing, has been over the many years and it's just getting very big. It started out as a users meeting and became just a little bit more 04:30:00organized by the time I joined it in 1958. For me it was an opportunity to meet in person many people whose work I had read about in the previous one and a half years since many of them gave talks at the meeting. I could listen to and talk of course to a lot of CEC people about the instrument. CEC was never an application research entity. They were very good engineers, ion optical people, but application -- they left that to the users.

04:31:00That was another purpose of the meeting; CEC could harvest last year's 'crop' of ideas. They used it to get new customers and to pick up this or that modification, which the user had come up in his laboratory because he needed it. I learned about most of the people, and could ask all kinds of questions. I told them about how I had just installed a mass spectrometer and they asked what I was going to do with it. When I told them what I was going to do with it they said that I was crazy. One doesn't do that. On the other side were those that 04:32:00thought it was interesting and they wished me good luck. I asked some questions that pertained to what I was going to do.

The next year it was in Los Angeles. It was close to Pasadena and so we visited the plant and met some of the plant people there. I gave a talk about peptide sequencing. People saw me and knew it was my second meeting and noticed that I was already giving a talk, which was unusual. I have given talks ever since. 04:33:00That was one way of showing to that group of people what one can do. My very first paper on peptide sequencing was a brief communication, just one page in JACS [Journal of the American Chemical Society]. I'm sure nobody in that group read it. The alkaloid and some of the alkaloid things were published in Tetrahedron Letters which was almost exclusively a natural products journal for fast publication that people in that group never read, or in Biochemical, Biophysical Research Communications that they also didn't read. So, the meeting provided me the opportunity to bring directly to that community what else one 04:34:00can do with mass spectrometry. And this surely happened with other people coming in from the outside and rounding out the program of the meeting.

Then it became so large that it became an incorporated society and became the ASMS conference. At this point, the last one I attended was in 2002 for the fiftieth meeting. Even before, the program was getting, to a certain extent out 04:35:00of hand, because there are so many things going on in parallel. Everyone has to really make a detailed schedule of what papers to attend. Early on, the question was why not divide it into a biological meeting, an inorganic meeting, and an environmental meeting, and so on.

But I think one of the advantages of keeping it together is that if one wants and makes the effort, one can learn what's going on outside one's immediate 04:36:00field. And I think that's always very important because it's sort of a cross-fertilization process. It does take time and effort and energy to handle this meeting. Then there is the proliferation of posters, which nobody can possibly see, short of just walking by and I think 80 percent are unimportant. It's just a way to justify students and post-docs to attend meetings, so they 04:37:00can charge it to their research, which is sort of an unfortunate formality. In the 1970s or 1980s for a few years there were rules that from each laboratory you can only have so many presentations. Since those are counted by the head of that laboratory whose name is on it, you could easily avoid that rule by just leaving your name off five papers that exceeded the magic number. Those rules were hard to enforce and didn't make too much sense. But I think nobody has come 04:38:00up with a better idea, and of course there's always the argument that give the students and post-docs exposure and people can associate a face with the name.

GRAYSON: It's an opportunity to meet the people.

BIEMANN: You can ask some questions, and sometimes people even get into heated discussions around the poster. But that's fortunately only in a few instances, 04:39:00and it never became physical, but you just have to take it as it is unless somebody comes up with a better way of doing it. But splitting it up into smaller separate meetings would lose that cross fertilization aspect. Of course, one could then go to all of those meetings or at least some of them and that's a possibility. But, anyway, it is a very useful thing, and one just has to manage it.

GRAYSON: It's not perfect but it's the best thing we've got, right now. I know that the program committee does review the abstracts now, and does actually 04:40:00reject some.

BIEMANN: Only a small number are rejected, mainly on technical grounds because it's hard from the short abstract to tell whether it's an also ran or a bombshell, particularly if the person who writes the abstract isn't well-versed in abstract writing. That's a negative aspect which the annual conference has is 04:41:00on the resulting annual cycle of work and manuscript writing in this field, because in the two weeks before the abstracts are due everybody is busy getting some final data which you can just scrape up and write an abstract for it that doesn't commit yourself to the impossible. But sometimes it comes close to it.

Then there is a second peak, the few weeks before the meeting where everybody scrambles to get the data that were promised, plus a few which were not in the abstract just to have a little bit of frosting on the cake. Then you have to 04:42:00create the posters, which is to a certain extent much easier because of all the computer and imaging systems that are available, but also more demanding because now it has to look good, because you have to attract the attention of the people who just have the time to walk by. The people have to be caught not only by the preliminary abstract, which is in the book which you get two weeks ahead of time, but also by the graphs and pictures in your poster. After a few weeks of recovery, now you can write the manuscript for publication.

04:43:00Of course, before you get to the meeting you have to write the long abstract, which is the intermediate stage. Then you feel you have to publish it because now that everyone knows what you are doing, you better not be caught by somebody who's doing similar thing, like the three posters down from you. Once those things are published, you really can't say I did this, but I'm too late. So that's one of the scientific management aspects of the annual meeting where everything happens at that time, and all the deadlines are created.

GRAYSON: They do have the Sanibel Conference, the Asilomar Conference and what 04:44:00they call Fall Workshops, which are more focused topic wise. These are all small group meetings. They probably have more of the feel of the early conference. I think that's what the society is doing to try and compensate for the fact that the annual conference has become like an ACS meeting. These smaller conferences are more congenial and people interact better.

BIEMANN: I only went to two Asilomar Conferences I think. And two Sanibel, one which I chaired and the other one the year before to figure out what's going on. 04:45:00And of course they were all very interesting but if I remember correctly, the number of attendees is limited.

GRAYSON: They tend to limit them.

BIEMANN: I think it's somewhere along the line of 100, which then lends itself to an in-group mechanism. Depending on the topic, you have to invite certain people who are well known to work in that field, the leaders in that field. And 04:46:00then you have to find the people that maybe are coming up and then you have to get in some students and post-docs and those from other laboratories. With the Asilomar Conference, I don't know the management connections, they just invited me to give a talk. In the first Sanibel I went to see how it's run, and in the second one, I was involved in helping run it.

GRAYSON: Well, I think we've covered pretty much the area that I was interested in trying to talk about. As I indicated at the beginning there's a reasonable amount about your career in the literature that you know is easily accessible. I think this is good because it gets a little bit more personal and down to 04:47:00details that you do not normally publish. This is the philosophizing part of the interview--you've had a career, a very high-powered scientific career, and in a dynamic field. And what does the future look like to you? Where do you think mass spec in particular and science in a broader sense is headed?

BIEMANN: I think mass spectrometry now is obviously about usability to a certain extent, because the more widely it's used the more routine it becomes. Also the 04:48:00pioneering number of events will be the same as it was 50 years ago, and it shouldn't be judged on that part. As we have seen since 1981 when Mickey Barber developed fast atom bombardment ionization, the advances in mass spectrometry have always been caused by new ionization techniques. Even before there was a 04:49:00number of things; chemical ionization of course, and californium plasma ionization, field desorption but they never really caught on because they're too difficult to do and didn't really work that well, or weren't really applicable to many things. Ionization techniques really have opened and enlarged the applicability of the field, and made it even faster.

Computers ran in parallel with that and played a great role in the automation of 04:50:00the instruments. The faster and more easy to use the instruments became, the more data you could generate; particularly in the automated ways that you now can run mass spectrometers. The amount of data generated could not possibly be handled without modern computer techniques. In that respect mass spectrometry without computers wouldn't be where it is now.

That applies to the application on a broad basis in many laboratories where it's 04:51:00just used as a routine instrument. It is hard to say if that will continue at that pace because there's only so much data you can use productively. What it does is it makes the operation of the instrument and the use of the data more generally useful, but it has the danger that the data are generated, produced and consumed by people who don't have the experience to do it right. Now for 04:52:00routine things, that's probably okay because routine means that you do the same thing often enough that it's hard to come up with the wrong answer because if it's really wrong, it would be such an outlier that it would be recognized as such. So, that is a problem that one always has to worry about.

Science in general depends on what new instruments will be developed and what new uses they will find. For example, DNA sequencing, thirty years ago in 1977 04:53:00-- next year will be thirty years -- the first DNA sequencing method was published. It was very tedious; very error prone. But then, when the human genome project push was on, that provided an impetus to do something to handle the enormous amount of analysis that needed to be done. People at Applied Biosystems and PerkinElmer came up with a new instrument, made 250 units and gave them away to Craig Ventnor's company. They -- just by brute force -- solve the problem swiftly. And here it is. But now there is only nibbling beyond that 04:54:00in other genomes, of other organisms. But the big thing is done. Then the interest always kind of falls off a little bit after something like that.

Certainly computers are important in every respect -- starting from designing an instrument for production, computing ion beams, and then testing the instrument, automatic testing of instruments twenty four hours a day to make sure the parameters are all right when you come in, in the morning and you push a button and it's going. The running of the instruments, the interpretation of the data and distribution of the data, relies on computers. The database searching, that 04:55:00is on the internet. The only trouble nowadays is that the internet is just greatly abused by business, on one side and criminals on the other side, which will eventually lead to a crash of the system. But, it certainly is part of the rapid development of instrumentation and the development of techniques and methods in all areas of science.

Now whether mass spectrometry is just one of them or for one reason or another unique, I don't know but I really don't think so. I think it had its great 04:56:00exciting times but also in the past some unexciting times; in the 1970s for example we were in a period where not much was happening in mass spectrometry. The instruments were refined, pushing the resolution.

GRAYSON: That was the biggest thing for a while.

BIEMANN: Pushing the mass accuracy was the big thing, but nothing much else happened. To a certain extent, I was lucky because I could devote some of my time to the NASA project, moon and Mars while my laboratory was running smoothly and people worked on the peptide sequencing, refining the methods. But then as 04:57:00we already discussed at length, the jump came with fast atom bombardment.

I don't think any comparable thing happened in the last ten years; comparable to how FAB changed the field and what electrospray and MALDI did -- and the instrumentation aspects that went with it. The driver now is to identify more and more proteins in fewer and fewer spots on a two-dimensional gels with less and less material and finding more and more modifications of those known or 04:58:00predictable proteins. It's a very important thing, no doubt about that.

But it's now at the stage where you need to accumulate more and more information to round out the biological system to understand all the things that go on in the cell at once or consecutively, which is very important. Now mass spectrometry becomes an automatic tool, semi-automatic tool, not quite automatic. You still have to do lots of things with your own hands, with pipettes. But it will be interesting to see what the next thing is that comes 04:59:00along in general. Not just in spectrometry. And often people ask me why MIT did not continue in mass spectrometry. Some people think that MIT closed my laboratory by decree. But that's not at all the case.

When I was asked by department heads and deans what to do when I retired, I said find someone who is going to work on a field that was as new as mass 05:00:00spectrometry when you hired me. Because that's what an academic institution should do with their faculty, move things ahead, and not just continue the things that can be continued just as well elsewhere.

It may be that other factors, unrelated factors, like renovating of the building that my laboratory was in had some effect. I didn't even tell you that where I moved, two years later that building was completely renovated and I had to move at least my office temporarily to another building and then back -- but at least not the laboratory because it's easier to close it than to try to keep it running.

05:01:00GRAYSON: Are you familiar with this ion mobility spectrometry that's being developed by a handful of young people?

BIEMANN: Yes. And I understand that the person who got the Biemann Medal this year is in that field.

GRAYSON: Yes, David Clemmer I think.

BIEMANN: I was told the name and I sent him a note, but I don't remember it right now. Of course, ion mobility spectrometry is nothing new. But then, mass spectrometry was not new in 1958 either. It just took a new twist. I don't know what those people are doing with it.

GRAYSON: It's a tandem instrument in that you get the ion mobility and the mass spec so you get some information about the--

BIEMANN: The three dimensional--

05:02:00GRAYSON: . . .molecular conformation with ion mobility and then you get some information about the primary structure at the mass spec end. It's a technique that probably would fit in the category that you're defining, that MIT should find. It's nothing new but the way it's being applied and implemented in the biological community is pretty interesting. That would be an ideal starting point I think. But I'm just curious because it is a really interesting idea. We've got a couple of speakers coming in to talk to our local discussion group about it. It seems like it's going to be quite powerful.

I don't know if there's a whole lot more to cover. We've covered a fairly broad range of topics including most of your career I think. You never really had any quadrupole instrumentation in your laboratory?

05:03:00BIEMANN: We had a quadrupoles for a short period. It was when Guy Arsenault was in my laboratory. You can look up in the MIT school of mass spectrometry paper for the spelling of his name. In fact, all the names of my graduate students and post-docs are there. But we only worked on positive/negative ion one after the 05:04:00other with the quadrupole. I think that was it. But quadrupoles never reached the performance of magnetic instruments, except speed. So, there was no incentive for me to get involved in a completely different type of instrument; 05:05:00except with Time-of-Flight mass spectrometers in the 1990s. But that was because they had particular performance characteristics which quadrupoles in the early times didn't have compared to other instruments.

We had lots of instruments; of course, we had the first Time-of-Flight mass spectrometer, a Bendix system, as we already discussed, which I got in 1961 or 1962.

GRAYSON: Did you ever go to any of those Bendix symposia conferences that they used to have?

BIEMANN: No. Because again, it was used sort of on the side for things which we 05:06:00tried out and looked promising and then moved to the magnetic instruments.

GRAYSON: So it was kind of test bed instrument?

BIEMANN: Where we couldn't do much damage.

GRAYSON: You couldn't hurt the instrument. So, you tried some weird things and if it looked like it would work, then--

BIEMANN: The only tangent connection with quadrupole mass spectrometers I had was during the Viking project when we needed to see whether the quadrupole instrument would be a better mass spectrometer for a flight instrument. Bendix Aerospace built a miniaturized prototype of one. But it turned out that the 05:07:00power requirements were greater for that than for a magnetic instrument using a permanent magnet. The performance of the magnetic instrument, being a double-focusing one, was much better than the quadrupole which would have required more power. But, they did build one and we looked at it carefully. One of the most important reasons for actually building it was that project 05:08:00management required it. At that time in the early 1970s everyone on the project management was saying that we had to use the quadrupole because it's so simple.

The project management wanted to make sure that they couldn't be blamed if something with the magnetic instrument didn't work either before flight or afterwards. But it was built and tested. Now almost all the mass spectrometers that fly in space are quadrupoles, even the ion traps. And I never had anything to do with ICR [ion cyclotron resonance] or FTMS [fourier transform mass spectrometer].

05:09:00GRAYSON: The Bendix Time-of-Flight was used quite a bit or at least looked like it would have been a good instrument for GC-MS. It had the speed in terms of the scan speed.

BIEMANN: Yes.

GRAYSON: But it was eventually beat out by quadrupoles?

BIEMANN: And the reason was that the early Time-of-Flight mass spectrometers, and the Bendix was the major commercially available one, had relatively low resolution. When it came out there were no recorders that were fast enough to 05:10:00record the signal. The electronics were of the 1960s or even 1950s. Gohlke used it in McLafferty's laboratory to more or less show that you can record and see the mass spectrum of toluene while it comes off the GC column.

The mass identification was more or less measuring off the oscilloscope screen using a Polaroid camera. So if you know what it is, you can say, this is mass 92 and mass 91. But when it is completely unknown it would have been more 05:11:00difficult. The only advantage it had over magnetic instrument was the speed but it was almost too fast. It could also tolerate a large gas load which with GC-MS was the main problem to overcome before separators were developed. As history shows, aside from that one paper, maybe two, that was it. There were a number of other ways to try to get mass spectra of GC eluates: by complex trapping and 05:12:00then injecting methods, but they never got beyond the demonstrations that it can be done.

GRAYSON: It was too cumbersome. I'm sure there are probably twenty or thirty methods of interfacing an LC to MS prior to electrospray. But everyone had their own method they were trying to develop and once electrospray came along it was like problem solved, and all the other stuff was cast aside.

BIEMANN: Then there was the big comeback of Time-of-Flight mass spectrometry when MALDI was developed because it needed a pulsed instrument. But at that time, the Time-of-Flight mass spectrometer had been around only since Bendix. They were steadily improved for very special things. But then that ionization 05:13:00method made it necessary to build a high performance instrument of that type. By that time electronics and computer control had advanced so much that it was relatively easy to build a high performance instrument almost overnight, and it has been improved since.

For example even at that intermediate time Mamyrin in Russia had developed the reflectron. Nobody was using it, except him and a few other people, until MALDI 05:14:00came around and it was found of great use. So now all MALDI Time-of-Flight mass spectrometers are reflectrons in one form or another.

GRAYSON: I started using Time-of-Flight instrument when I was at McDonnell-Douglas Research labs when I first started there, that's what they had. I've always been fascinated with the fact that today you can get a Time-of-Flight instrument with resolving power of 10 or 20,000 and back with a Bendix machine a couple hundred resolving power was the most you could get. It's just fascinating to me to think that you could improve the technology of that instrument, that analyzer, so much that there's two orders of magnitude improvement in the resolving power, due primarily to developments in electronics.

05:15:00BIEMANN: Faster electronics and computer control of all the parameters down to high limits made it possible. While on the magnetic instruments computer control wouldn't have done much to the performance. It was the ion optics that had to be tuned. Of course computers helped there by calculating the important parameters. First, Matsuda and then Matsuo, his student, refined ion optics for magnetic sector instruments to achieve higher resolving powers.

05:16:00GRAYSON: So the Nobel Prize was awarded to Tanaka in Japan for his work on MALDI. I think a lot of people in the mass spec community felt that Hillenkamp and Karas should have been awarded that. Do you have any thoughts on that?

BIEMANN: I completely agree with that. I can't understand the notions behind that decision. The only one that could cause it was that he is not a European or American, but to give it to a Japanese once in a while, which of course doesn't really carry any water. But, I think there were some people who say that Tanaka 05:17:00had the first experiment successfully in that area, and presented it at a meeting in Japan. Many of the advocates of his priority claim are people who attended that meeting and they were quite impressed. But, I think for the Nobel Prize, you have not only to show who published the first paper but also whether he actually did it right and whether it had any practical usefulness and I don't mean in a commercial way but in a scientific way.

The thing that I would fault him on was that he did not disclose what particle 05:18:00size and I don't think he was quite clear whether he used palladium or platinum. But certainly, he did not tell us the particle size, which is the most important parameter of that method. That made it therefore impossible to reproduce and use it in other laboratories. While Franz Hillenkamp laid it out in great detail how to do it and therefore we all could do it. I think since they gave it to Tanaka, they should not have given it to John Fenn, but to Malcolm Dole, who did the 05:19:00first experiments twenty years earlier. I don't remember whether Dole was still alive.

GRAYSON: That is a problem.

BIEMANN: If he wasn't alive, he couldn't get it. Dole published more details in his paper than Tanaka did in his, but he didn't get anywhere because they didn't know what was going on, nor did Tanaka and he didn't carry out the necessary experiments to figure out what was going on. I know that in quite a bit of detail because I was a member of the NIH Study Section when he applied for a grant to figure out, and develop that method and do it right and show what it can do.

GRAYSON: He being?

05:20:00BIEMANN: Dole. He was at Northwestern University and was going to retire and move to the University of Texas. The review committee was quite convinced that it's an important thing, very important if it works. Since he was sort of doing the experiments and making the measurements -- he used polyethylene glycols as 05:21:00molecules to measure the molecular weight -- which he might be able to do it and if it worked, it would be great. But since he is going to retire, and he's going to move, the chances are low that he would make it work. It was decided to fund his application because of that slight chance that it might work, and it would be important if it did work, so it was worth some money to fund it. But of course, he moved and then never did anything else.

GRAYSON: What was the year?

BIEMANN: I don't know.

GRAYSON: Around the 1970s?

05:22:00BIEMANN: Either in the late 1960s or early 1970s. It was around the time when he published those two or three papers. I probably could find out. But then it took someone like John Fenn who came from the other side, mainly the engineering.

GRAYSON: The nozzle design, engineering . . .

BIEMANN: . . . the molecular beam side to figure out what's going on. Sandy Lipsky who was at Yale bumped into Fenn some time after he moved to Yale. Lipsky 05:23:00had read Dole's papers and suggested to John Fenn that he look into it. He might be able to understand what's going on. So Fenn looked at it--

GRAYSON: So Lipsky interacted with Fenn in passing?

BIEMANN: Yes. They never collaborated because Lipsky was in the medical school.

GRAYSON: Fenn was in engineering.

BIEMANN: It was by accident that it happened -- like when I listened to the papers at the flavor conference, and found out about mass spectrometry.

GRAYSON: So one has to take advantage of these opportunities and make the most 05:24:00of them whenever they occur.

BIEMANN: John Fenn could have said, "I'll look into it" and fifteen minutes later forgotten about it. It was a coincidence and then he recognized that there is something that is worth looking at.

GRAYSON: Science is really a gray, non-linear process in many ways. I think the average person just thinks it progresses in a very natural step-wise fashion but it's a very tortured path, and then little things mean a lot.

I've covered I think pretty much of the areas. If there's anything else now's a good time, to leave your thoughts to posterity.

05:25:00BIEMANN: Something I think about lately is planetary and space research. During the Viking project and before that the Apollo, it was all new and very exciting. You never knew what you would find and you had to keep an open mind. All those factors in addition to the fact that there was money available, for one political reason or another, to do those very expensive things. They had much 05:26:00scientific appeal but much more public appeal which made it possible to scrape up some money. But there was no NASA community. So people from academic institutions and independent or government research organizations were drawn in to do it more or less. None of them had a particular ax to grind. None of them had their scientific livelihood and research funding depending on the project 05:27:00because whatever they did was funded by something else before NASA existed.

The people who got together and volunteered or were drafted to do it, were enthusiastic about it, had nothing to lose, had no ax to grind, and just did it going along with their other work. The Viking project was organized, the experimental science aspect was organized, into teams of scientists for each experiment; and they came from all over the place. On the GC/MS team, one came 05:28:00from the University of Texas, one from the Salk Institute, one from the Geological Survey in Washington, D.C., one from the Low Temperature Research Laboratory here in Hanover, New Hampshire and one was an astronomer from Stony Brook.

We met in person because on the telephone it was hard to do. There was no E-mail. So we just had to get together. We talked things over. One of them was from JPL. A second one was also from JPL who then moved to the UK during the 05:29:00design of the mission. So we had to meet in general at JPL or maybe once or twice in Washington. Once we met at this house here. We had to get together for a few days. None of us knew each other before. It became a very congenial group.

Then there was a science steering committee, which consisted of all of the team leaders where those things were discussed where everybody was affected by it. For example, the data allocation was a big thing, particularly before that wire 05:30:00recorder was brought on board. That lasted from 1969, through the mission at the end of 1976.

The science steering group got to know each other very well, not socially so much, but in terms of science philosophy. The team leader of the imagery on the surface said he needed a picture at that time, on that day, three years from now and therefore we couldn't run a GC-MS experiment on the same day because it 05:31:00would be too much data. Then I knew exactly why he thought it was important to do it on that day, and he knew exactly what we were giving up, and vice versa. Anybody who didn't fit into that way of doing and mutual understanding fell by the wayside in the meantime and was just, either voted off the team or resigned or just was kept on the list but never attended the meeting and for all practical purposes wasn't there. That was from a human and philosophical and scientific aspect.

Maybe what I took away from the Viking project personally, particularly since we 05:32:00didn't find anything, was the fact that not finding anything had no affect on me at all. We went there to find out if there was something and if something is there, what it is. But even to find that there is nothing, at least in the top ten centimeters at the two places we went to; to me that was a full success. But sometimes people thought that I was terribly disappointed that I didn't find anything. And I said no. We answered the question.

GRAYSON: You were just elated that the experiment worked.

BIEMANN: My career wasn't depending on it. That brings me to the opposite side of the situation, which exists now. Now, those missions are run by NASA or ESA 05:33:00[European Space Agency] and the experiments are done by people whose whole career and research funding depends on proving some preconceived notion. They are all government scientists and engineers or are in departments at universities which are solely directed to what's called space research. For them it's a catastrophe if the instrument doesn't work because there was no power; 05:34:00that's one thing. Then it's embarrassing of course to the people who built it. But in spite of all that many of the projects are doing well, and finding something useful and reliable.

But on a recent mission a probe was sent through the atmosphere of Titan, the largest moon of Saturn, and landed on the surface. It happened on 14 January 2005; about one and a half years ago. Things went reasonably well. There is an instrument that is supposed to look at the chemistry of the haze particles in 05:35:00the atmosphere of Titan -- which is a big mystery, why there is haze on Titan. It was discovered first in 1980 by a Voyager fly-by. So in 1997, the Cassini mission was launched to Saturn and it took the Huygens probe along and then dropped it in the atmosphere.

There was an experiment that was to determine what those haze particles consisted of. In the twenty-five years since Voyager discovered not only the haze but also that the atmosphere consists of mainly nitrogen and a few percent 05:36:00of methane, lots of people have tried to do the Titan equivalent of the Miller-Urey experiment on earth -- which as you may remember sort of shed light on how life on earth got started. But, it was a very famous experiment and of course Stanley Miller got famous and Harold Urey was already famous.

Everybody and his cousin were irradiating over the last twenty-five years mixtures of nitrogen and methane with all kinds of things from UV to microwaves 05:37:00and electrical discharges to particle bombardment. One of the first ones was Carl Sagan at Cornell. In all those experiments a solid deposit formed if you waited long enough. This was an amorphous goo, slightly yellowish brown in all those cases. Carl Sagan with his flare for expression, since he was the first one who made this stuff, or at least the first one who felt that one should give 05:38:00it some name so that we can talk about it -- he called it tholin. It comes from the Greek word for mud because that's what it looked like to him, and what it actually kind of was, not mud in my driveway but mud chemically speaking.

Over those twenty-five years this group of planetary scientists -- some people call them Titan-ologists because they are concerned about what is happening on Titan -- became convinced that that's what is in Titan's atmosphere. One knows that the atmosphere consists of nitrogen and methane, which is irradiated; when methane gets irradiated, it forms ethane; and eventually forms acetylene, which 05:39:00can polymerize and the nitrogen gets involved in it. But nobody ever bothered to analyze what that gunk is. They did carbon/hydrogen analyses, and nitrogen, and they came out all over the place depending on what you irradiated and what temperatures and pressures you used in the experiment.

Recently a group in Texas did a FT-ICR MS measurement on such a tholin and got the elemental composition of all that stuff. It turned out that it's about at least one hundred and eight different things, as far as elemental composition goes; not even counting isomers. It's fifteen series of different 05:40:00carbon-hydrogen-nitrogen compositions. But nobody ever tried to separate it or do anything but run an ultraviolet spectrum of tholin. But people were sort of convinced that this was there and called it "Titan" tholins.

That experiment was designed by a group in France because it was an international effort, mainly run by the European Space Agency. The Cassini mission was mainly run by NASA and JPL. The Huygens probe to Titan was mainly 05:41:00European with some Americans involved in it. The mission was to collect the sample, put it in an oven, treat it at ambient, 250 degrees, and finally 600 degrees (Celsius) to eventually pyrolyze what didn't come off before at the lower temperatures. In that respect, it was similar to our Viking experiment.

What comes off is introduced into a mass spectrometer, which was flying on the mission to analyze the atmosphere and what happens on the surface. By the way, the surface of Titan at one time was supposed to be an ocean of liquid ethane and methane, which was later reduced to just lakes, and oceans. There were people that worried that the probe could sink when it gets to the surface.

Great emphasis was therefore on what happens before it touches down. That 05:42:00experiment just collected those particles in the atmosphere then heated them up and what they found in the mass spectrum was ammonia and HCN. The title of the paper published in Nature on 8 December of last year was "Complex Organic Matter in Titan's Atmospheric Aerosols by in situ Pyrolysis and Analysis."

Basically they found complex organic matter making up those aerosols; but only 05:43:00by detecting ammonia and HCN. When you read that paper, it doesn't really say what they mean by complex organic matter. But then since it's a "Letter" in Nature they can't put everything in the printed form because they have to be short, so there is supplemental material that you can get only online.

I read the paper, and I said, "That's funny, just from ammonia and HCN, how can you get to complex organic matter?" It was in January when I got to my office -- I went to the library and looked at the hard copy, and I noticed that there was 05:44:00supplemental material and there were six figures in it and the last figure is the structure of that molecule. It is a very detailed structure which looks almost like a large steroid; it has an aromatic ring and a cyclohexane ring and is connected with aliphatic side chains which have various methyl and ethyl groups on them -- and then, hanging off, an amino group, two imino groups and two nitryl groups. So that's where the ammonia and the HCN comes from when you heat it and you pyrolyze it. I was so flabbergasted to see that somebody can 05:45:00draw a structure of that detail which, on Earth would take you at the minimum two months of detailed analysis but probably a year or so, that one can conclude that from ammonia and HCN. But it was published. So I read it in detail and it shows three figures of mass spectra collected from two samples. They showed the 600 degree pyrolyzate for sample 1at 130 kilometers altitude; sample 2 at 25 to 20 kilometers altitude and then another figure that shows the mass 17 and mass 05:46:0016 signal for 18 spectra, which they collected while they flushed the pyrolyzate into the mass spectrometer.

The mass spectrometer had measured the carbon isotope ratio in the methane of the atmosphere and calculated that ratio. So I measured on the paper, enlarged on the Xerox machine, the peak heights and it turned out that the entire mass 17 signal with the exception of one scan is accounted for by the required 13C methane signal, so there is no ammonia. They said in the legend that for sample 05:47:001 the results are similar. In other words, there is no ammonia. And then they say that in sample 1 mass 27 (for HCN) is the same as in the background and in the other sample it's a little bit. But you really don't know whether that is from all HCN or whether it's a fragment from ethylene or ethane, which you can't tell because there's so much nitrogen at mass 28 and its isotope at 29. There is a big peak at mass 30 because it's from the flush gas -- they used pure 15N2 to differentiate it from Titan's atmospheric component. So, there's no evidence for 05:48:00ammonia and for HCN but they say in the title they have discovered complex organics; and to show what they mean by complex organics they show that structure of a "tholin".

As another proof they show the pyrolysis gas chromatogram of the pyrolyzate, which they had produced in their own laboratory. That's a sixty-minute chromatogram, but they cut it off at 10 minutes to only show from two minutes to ten minutes to show that ammonia and HCN is produced when they pyrolyzed their terrestrial junk (tholin). So that's the other proof of that finding. There are 05:49:00two U.S. authors on it; the one who did the GC-MS and the atmospheric experiment, and the other one is a good friend of mine -- an astronomer who was on the Viking project doing the atmospheric analysis and isotope distribution of the noble gases.

I finally called him up and asked how he came to that information, and the mass spectrometer person said, "I just gave them the data and that's their interpretation and so they told me they found ammonia and HCN, therefore there are complex organics there." He was worried about it, and how they got to that, 05:50:00and I said (there are 22 authors on it), "Why did you leave your name on it?" He said, "I tried to take my name off, but it would have caused an international incident because it was a collaboration of the European Space Agency and NASA in the U.S." Of course, my astronomer friend had even less to do with that interpretation.

But I'm now publishing a rebuttal of that in "Nature." They agreed to do that but I haven't cut it down to seven hundred words yet. Nature said they won't publish it unless I can do that. It turns out that those people for many years didn't do anything but work on their ideas that those compounds must be there 05:51:00because we can make it in the laboratory. Then on top of it is the fact that they say that the ambient temperature and the 250 degrees didn't get any data because the gaskets which they used to close the oven up weren't soft enough to close at those temperatures because they were very cold through that interplanetary flight. Therefore it leaked, there are no data for those temperatures -- that part didn't work. The GC connection didn't work either. So they only have those two 600 degree pieces of data.

I wrote to them first. But they are so convinced that since you can make it on 05:52:00earth it has to be there, that they just believe it. None of them understand any organic chemistry apparently. In France it's all government laboratories and they either didn't realize that it didn't work or can't admit it or aren't allowed to admit it because the marching orders of the European Space Agency was if we give you money--

GRAYSON: It works.

BIEMANN: -- for that experiment, you better publish some data. Not that I want to say NASA tells people to publish or perish a terrible death even if it's 05:53:00wrong. But it's kind of the same idea; because the people have been in that business for the last thirty years, so they are very entrenched and are close to retirement. Therefore, they think more about their retirement pay than the experiments that they do; and that is somewhat unfortunate. It's very different from the situation thirty years ago when there were no preconceived notions of what was on Mars because we didn't have twenty-five years to dream about what could be on Mars and therefore had to be on Mars.

05:54:00GRAYSON: Careers didn't depend on the results.

BIEMANN: It would be good if something could be done to inject fresh independent blood and even more fresh and independent thinking into that system than to do things that way now.

GRAYSON: So you sound a little bit like the editor of JACS [Journal of the American Chemical Society] who said where is the melting point data? But you have better proof in your case where you had real hard information in front of you. But now these gentlemen are trying to say that there's something there because they see--I mean it's really hard to believe that they only saw those two compounds and they're going to say that represents a complex organic--

BIEMANN: On top of it, if what they say is not so.

GRAYSON: Yes, well I remember you sent an email asking for the relative 05:55:00abundance of the peaks in the methane spectrum. I'm thinking, "What does Professor Biemann want to do with methane?" That's an interesting commentary on the state of science.

BIEMANN: Of course one has to say that in that field you can't do one experiment today and another one tomorrow, and a third the day after and decide which one of the two were right, you wait another twenty years until you can do it.

GRAYSON: They need another career.

BIEMANN: You have to do it step wise and at each step just add the real fact. Not . . .

GRAYSON: . . . imagination.

BIEMANN: One of the reviewers of my rebuttal said it is important to point out 05:56:00that the original paper is incorrect because if it stands, then for the next twenty years people will base their terrestrial experiments on that. It wouldn't take long until one of the new brands of astrobiologists would say that we just have to move fifteen bonds around and we come up with something like ergosterol. Therefore there must be life on Titan, even at 98 degrees Kelvin.

GRAYSON: I think that is very good--

BIEMANN: Enough of philosophy.

GRAYSON: Yes, well that's a problem because there's this gentleman in Germany who got caught publishing something about organic semiconductors about five or 05:57:00six years ago, and he was able to show that he could create organic compounds that would behave as semiconductors. Obviously, this would be something pretty tremendous but then people tried to reproduce the work but it wasn't reproducible and eventually all his papers on the subject were withdrawn. But it's unfortunate that people get involved in these situations where they feel that their interpretation of something is greater than it really is.

BIEMANN: People can easily get caught in an idea and follow that and don't see the red lights flashing left and right because they only see the green light 05:58:00ahead of them, and don't realize that it's a reflection from the back. I always say that the horse is an animal with four legs, but not every animal that has four legs is a horse.

GRAYSON: Yes. I just wanted to make you aware of one comment that I read recently in the "Scientist". A gentleman at the University of Pennsylvania by the name of Blair who's involved in their proteomics effort, and there's a quote in this article in the "Scientist" where he says that "Mass spectrometers are like handbags, you can never have too many." So, that's a pretty interesting attitude about mass spectrometers that you just want to have more and more and more of them.

But I understand -- Dr. Gross consults with some of these pharmaceutical 05:59:00companies--they try to have one high performance mass spec per two investigators in the pharmaceutical research lab. So you can imagine that there are a lot of mass spectrometers in the pharmaceutical business. That whole idea of never having too many comes from that kind of attitude or approach. It definitely is a technique that has burgeoned in the last fifty years and to a large degree, your work has been an important part of that actually happening and being what it is today.

Thank you very much sir. I'm glad we were able to do this, and I hope that your wife is feeling better. Can I take a couple of quick pictures before I leave?

BIEMANN: Let me comb my hair.

06:00:00GRAYSON: Okay. Sure. You have a word list for me?

BIEMANN: Yes, but now it's outdated, but it's probably only a quarter of the names we mentioned and I can expand it from my memory.

06:01:00GRAYSON: Okay. You'll have an opportunity to see the transcript and we'll be able to read through it, and then it should prompt you also for the names that we didn't get.

BIEMANN: But I can send you that expanded list by email.

GRAYSON: Okay, that'd be great. Very good. You want to grab a seat there and I'll just take some informal pictures while we're chatting a little bit. And so you're going to a conference in, this October is it.

BIEMANN: Yes, end of October.

GRAYSON: And that one's going to be on the--

BIEMANN: I think I sent you the copy of that manuscript about laying the groundwork of proteomics.

06:02:00GRAYSON: Yes, that's going to be kind of a retrospective--

BIEMANN: Retrospective.

GRAYSON: Is this a conference of protein chemists?

BIEMANN: Proteomics.

GRAYSON: Oh, that's the HUPO conference. That's kind of an awkward sounding name; HUPO. I believe that Judith Sjoberg is actually doing that conference now.

BIEMANN: Yes, I just got her email with the layout for the program. It mentions about the organization and gives an address in New Mexico, and I thought it's her.

GRAYSON: Yes, it is the same organization. She apparently has just starting doing that.

BIEMANN: Of course, it's logical branching out of mass spectrometry.

06:03:00GRAYSON: Yes, she's been very effective in getting these things going real well, and then she's really been great I think for the society (American Society for Mass Spectrometry). I think she's done a really good job and helped in many ways to expand and make it a better conference.

BIEMANN: And I understand her son now works with her.

GRAYSON: Yes, Brent is working with her in that business. I think her daughter worked for a while with her but I guess all the details didn't work out. Her daughter is a pretty energetic person.

BIEMANN: Two energetic persons together sometimes doesn't work so well.GRAYSON: I should actually take some pictures out your back window if I could. So you moved out here in '98?

BIEMANN: 1998.

06:04:00GRAYSON: That looks so wonderful. Let me step out on the porch here.

[END OF AUDIO, FILE 1.3]

[END OF INTERVIEW]