Reminiscences of a career in mass spectrometry in the US and in Spain

This has been indeed a totally unexpected and surprising invitation from the Editors of Mass Spectrometry Reviews, and I want to thank them, as well as any other colleagues whom I suspect, may have had something to do with it, for brightening my retirement. After being retired for 10 years now I have no way of contributing a scientific manuscript for this issue. Instead, I think that it may be a good occasion to review my past professional experiences in Mass Spectrometry (MS), including those acquired in attempting to develop biological and biomedical MS after my return to Spain and compare them with my earlier experience in the United States. I could summarize the US experience as most satisfactory and unforgettable on all fronts while the years in Barcelona have been like jumping on a roller-coaster for a wild ride, full of unexpected turns and surprises, but challenging and unforgettable as well. Now, a full 10 years since my retirement, I feel that I can be candid enough and freely write on whatever comes to mind and expose some situations, experiences, and unexpected courses of events that I have not shared with anybody yet, not even family or coworkers. My professional reminiscences start on September 7, 1964, in Barcelona, when, after packing up all my dreams and plans, I boarded a PANAM flight to New York and from there the trip went on to Houston where I had been granted a teaching assistantship at the Chemistry Department of the University of Houston and conditionally accepted for Ph.D. work at the laboratory of Dr. John Oró. He was a prominent biochemist from Lleida, Spain, who years before had obtained his Ph.D. in Biochemistry at Baylor University in Houston and who, was interested in unraveling the mystery of the origin of life, had just demonstrated the abiotic synthesis of adenosine. I guess I should add here that my acceptance at the University of Houston was the culminating result of an unwavering resolve to achieve goals I had begun to define almost 8 years before. When I was 14–15 years old, I started studying English and after 6 months into it, I remember going to the US Library in Barcelona for English books. I came away with Tom Sawyer and Huckleberry Finn. A huge challenge, indeed! After having digested these, and many other books and magazines, I was already translating Western novels for a local publishing house as well as corresponding regularly with various pen pals in different US states. I was also thrilled by American democracy, its institutions, and its scientific and technological achievements. All of which finally coalesced into a firm will to continue my studies in America. I will spare the reader the description of the shock of moving from a gloomy and politically dull Barcelona to the fast-paced and booming environment of Houston in the early 60s. Lately, both my wife Esperanza and I have been enjoying the TV series entitled "Mad Men" for the accurate general and detailed depiction of life in the United States in those early years. The series is not about science labs but advertising agencies. However, the representation of the American way of life in the sixties is just what we remember (astonishing cars, comfortable and well-equipped homes, a free society with plenty of opportunities). Life in Houston was hectic and very demanding, both at the department and outside the University, but my English was proficient enough by then and so I managed to adapt readily to the new environment and use its facilities to my advantage. Therefore, in January 1965, on the occasion of the semester break, I flew back to Barcelona, got married, and returned to Houston with my wife. We were a couple of dreaming newlyweds projecting a family, a home, and a scientific career. A few months after that, because then research money was generously flowing to the Universities under NASA's Moon sample analysis grants and contracts, one of the first three commercial units of the Swedish LKB-9000 gas chromatograph-mass spectrometers (GC-MS) ended up in Oro's laboratory (Figure 1). The other two units, as I remember, went to the department of Biochemistry directed by George Waller at Oklahoma State University and to the Mario Negri Institute in Italy. No one in the lab was prepared or knew exactly how to best manage the new instrument, due for arrival in mid-1965, but I do remember that only 2 weeks after my arrival in Houston, in September 1964, Dr. Oró had already proposed that I specialize in this new analytical technique. I guess that to catch my attention and interest he told me that I could become one of the few world experts on it and asked me to focus my doctoral work on its applications in organic geochemistry, abiotic organic chemistry, extraterrestrial sample analysis (meteorites and eventually moon samples) as well as the possible environmental contamination of extraterrestrial samples, in general. All I knew of MS at the time was from a short paragraph in one of the chemistry textbooks I had been using at the University of Barcelona. Therefore, the pressure and responsibility I felt since I laid hands on the new GC-MS instrument were overwhelming and my initial productivity while accommodating to the new environment and responsibilities was perhaps somewhat lower than expected. This was to the point that during the first half of 1965 Oró summoned me to his office to warn me that I needed to shape up and get my act together if I wanted to continue at the University. With the help of my wife, my own determination, and the intensive lectures on Chromatography, Physical Chemistry, and MS, then taught by Professors Albert Zlatkis, Ralph Becker, and Gerhart Meisels at the Chemistry Department, I managed to keep things under control until I, very successfully, graduated in August 1968. I should add here that during my first 5 months in Houston, before I flew back to Barcelona, got married, and returned with my wife, Oró was like a father to me, always trying to facilitate and speed up my adaptation to the new environment. Until I got a car, he used to take me around, when needed, or even occasionally he let me use his. He also made me feel at home with his family. Later, with my wife Esperanza and our two Texan sons, both families got very close together. Nevertheless, at the same time he was very serious and demanding at work and I soon realized that I was not to expect any professional advantages from him, which is how I thought it should be. Undoubtedly, I was at the right place at the right time because thanks to the recent development of the jet separator by Ragnar Ryhage at the Karolinska Institute in collaboration with George Waller, the direct coupling of GC to MS just had been made commercially available. This separator was to me the real core of the LKB-9000 instruments because it allowed a significant number of high-efficiency separations and identifications of organic matter with GC-MS. Further, the collaboration with Professor Zlatkis, a well-known chromatographer at the Chemistry Department, paved my way to the use of the, then experimental, 31- to 310-m-long coiled stainless-steel capillary columns (Figure 2). These were heavy and demanded a good physical form to lift and place them in the GC oven. Some of these analytical runs lasted for 24 h or more. To reconcile work with family life, I found a method to cool the column, stop carrier gas flow at night and resume it in the morning without losing any appreciable separation efficiency. GC coupled to MS was not only indispensable to identify the dozens or hundreds of peaks eluting from a GC column, but the GC stationary phase bleeding was also very useful in practice. At that time mass spectra were recorded on UV-sensitive rolls of paper and had to be counted by hand. This tedious task was facilitated by the well-known column bleed mass peaks that helped to set the correct mass axis. Since early on, Oró had assigned me an office desk located in one of the laboratories at the department of Chemistry where I spent most of my time till graduation in August 1968 (Figure 3). The boxes full of rolls of mass spectra, my daily companions, can be seen on the large center table. There is no way I could explain the numerous unforgettable experiences and satisfactions from that period but one that comes to mind is the extensive work I did on the characterization of possible contaminants of the moon samples when they were delivered by the astronauts. We had to make sure that if any traces of organic matter were to be found on these samples, possible terrestrial contamination could be discarded. One of the many items I analyzed was tobacco smoke and for this purpose, I built a mechanical smoking device that "smoked" a full package of Camel cigarettes. The smoke was trapped in a cold finger and the horrible-looking residue left was analyzed with GC-MS (Gelpí & Oró, 1970b). Just the sight of this residue before analysis was enough to make one stop smoking forever when one realized that all this residue ended in the smokers' lungs! I always wondered why this residue is not shown to people that want or need convincing proof to quit smoking! Although by 1967, thanks to my new toy, the LKB-9000, I had become already well familiar with MS and chromatography as well as with their coupling, I was following with much interest the work from other prominent groups doing top MS research at that time. Names like Klaus Biemann, Fred McLafferty, Geoffrey Eglinton, Al Burlingame, J. Seibl, Ragnar Ryhage, among others, come to mind now. I mention this list because in that year, Dominic Desiderio, who had graduated from Biemann's lab in 1966, came to Houston in 1967 as an Assistant Professor of Chemistry at the Baylor College of Medicine. Only naturally, I was immediately attracted by his credentials and, as we both now remember, I visited him several times at Baylor to learn and discuss sMy with a real expert. By the time I graduated, I was thoroughly enjoying my work and family life. What else could I ask for? I knew our LKB GC-MS equipment to the last bolt and had mastered and completed all the labaratory procedures to become the most-prolific Ph.D. student and postdoctoral fellow in that Department, as publicly acknowledged later by Prof. Oró himself. In fact, I had obtained so much experimental data that after my graduation Dr. Oró exceptionally asked me to stay for 1 year to process and incorporate all of it into various manuscripts. In the 5 years I stayed in Houston (September 1964–July 1969), I obtained enough laboratory data and information to publish 27 scientific manuscripts in top-ranking journals (see Gelpí & Oró, 1967, 1970a, 1970b; 1970c, 1970d; Gelpí et al., 1968, 1969, 1970; Gelpí, Han, et al., 1970; Gelpi, Schneider, et al., 1970; Oró et al., 1967; Schneider et al., 1970). Some of these publications are still being cited today, and one of them continues to be cited rather frequently (Gelpí, Schneider, et al., 1970). I guess I should add that I was also prolific outside the Department because by the time I got my doctoral degree in 1968 we already had added two newborn Texans to the family. Although it was absolutely thrilling to be able to be part of NASA research programs leading to the Moon landing and to have access to the new techniques of MS coupled to GC for the analysis of trace organic compounds of biological and abiotic origin, it was not only the scientific and familiar aspects that became imprinted in my memory but also many of the events that have been said to have changed America at that time. I refer mostly to the social and historic events we lived through during our stay in the United States like the Vietnam war and the antiwar protests, the marches for voting rights, the race riots in various US cities, the assassinations of Martin Luther King and Bob Kennedy, and many other political and social events that bring to my mind a song like "7'O Clock news" by Simon and Garfunkel. More than its melody, it is its lyrics that bring goosebumps to my skin every time I play it because it brings memories of actual news heard on TV at that time. Given my earlier enchantment with America and its way of life, all this social and historic upheaval was rather disappointing to me at that time but even so I never imagined, not even in my wildest dreams, that 1 day I would see on TV, a mob assault on the US Capitol! The following year, after pondering the pros and cons of the offers I had for postdoctoral work elsewhere, including one for joining a NASA science team at its Houston headquarters, I decided to accept the position of specialist research chemist with Al Burlingame at the Space Sciences Laboratory in the Berkeley campus. Thus, our family of four arrived there on the 1st week of August 1969, just a couple of months after the famous Peoples' Park protests, brutally quenched then by Governor Reagan. The trip from Houston took about 2 weeks of driving through the most scenic roads and National Parks of the West and Southwest, pulling a U-Haul with all our belongings all the way to Berkeley. On that trip, I fell in love with the American West so much so that I must admit that, over the years and after multiple trips to the most scenic places of the west from Alaska to Southern California, I now know quite extensively all of the US West coast states, including the neighboring Rocky Mountain states and British Columbia, better than my own country. I have always been interested in photography and before coming to the United States, with Alfonso Gutierrez, a good friend of mine, I learned to process and develop all my rolls of film including color slides, so I guess it was only natural that in my last year in Houston, I bought myself a Hasselblad camera with wide angle and tele lenses. With this unique equipment, all the trips I just mentioned above were properly recorded and documented in my extensive photo collection. In fact, I paid for the camera by selling the rights of selected images to a photo agency in Spain (agefotostock.com). The children were then too young to know, but my wife and I will always remember the last thing we did the night before we left Houston on the morning of July 21, 1969. That was watching the moon landing on large screen color TV at a Houston friend's home! My leaving Houston meant that I was not to be part of the University of Houston team receiving lunar samples for study. Instead, I was at the Space Sciences Laboratory in Berkeley when these most special samples arrived and did help to process them for the public exhibit of one selected piece at the Lawrence Hall of Science (Figure 4). With all that personal and scientific baggage, we began our new life in Berkeley. Living and working in the San Francisco Bay area was a truly delightful and unforgettable experience because it gave us the opportunity to visit and enjoy the most scenic places in California, Oregon, and Washington. For me, it was also a very challenging and productive year. Al Burlingame put me in charge of a project to isolate milligram amounts of steranes and triterpanes from the Green River oil shale. I did not engage in much MS work there but concentrated on setting up an automated preparative system for the gas chromatographic isolation of selected steroid precursors (Gelpí et al., 1971). All this study gave rise to three more publications and so altogether, since my first paper in 1967 till 1970, I had managed to produce a total of 30 original publications. One of the often ignored or taken-for-granted factors that contributed to all this productivity was the well-equipped and efficiently manned mechanical and electronic and glas blowing lab support facilities at the US universities (Figure 5). These were crucial for the servicing and maintenance of instrumental equipment in Houston and in Berkeley as well as for the projects that required the setting up or development of new and complex instrument modifications like in the case of the preparative GC system, I put together at the Space Sciences Lab. Thus, in the laboratories where I worked, one came to feel that your work was crucial and well supported. For instance, when working at Orò's lab on a project on the abiotic synthesis of hydrocarbons by Fisher-Tropsch mechanisms (Gelpí & Oró, 1970b), I urgently needed a tank of CO which was not readily available from local suppliers but which the Department, through NASA channels, got flown to me in record time from I do not remember where. Until this point, it all had been about hard work and high productivity on my part within a very supportive environment, but my productive situation would eventually change in Spain in ways I could not even imagine at the time. Al Burlingame was very interested in my continued stay at his lab for a longer period and was already processing, through the university official channels, all the paperwork for my Fulbright US visa extension. Under the Fulbright program visas were limited in time and exceptions had to go through the US Senate. However, maybe because I was not confident enough on the success of that Visa extension request or maybe due to the drive and incentive of implementing in Spain the high possibilities and bright future of MS in Biomedical research, I ended up accepting the offer and tempting promises from the newly established Autonomous University of Barcelona (UAB). Thus, in 1971, once back in Barcelona, I joined the staff of the recently created Institute of Fundamental Biology (IBF) at the UAB where I was told that any contract renewal, unlike in other Spanish universities, would not depend on various candidates competing for one tenured position, as it was the rule in Spain and, with some exceptions still is, but only on my own scientific productivity. A few years later I found out that bureaucracy and false promises were going to be one of my biggest disappointments. For instance, it took 10 years for my Ph.D. to be validated in Spain, which prevented me to access a properly tenured position. In the end, I was forced to submit to competitive oppositions. I guess that the only autonomy that the new Autonomous University really had, was reflected in its name! I could mention here that all other promises to encourage my return to Barcelona were never duly fulfilled. Once at the IBF, and unaware of all the difficulties ahead, all my attention centered on building up a good research team in the field of MS applied to biomedical projects. Attracted by my US credentials and CV, several young predoctoral fellows asked to be enrolled in my newly formed research group. To cut a long history short I will say that, after many broken promises and delays, the new Hitachi RMU-6H GC-MS unit (Figure 6) that the university had pledged to buy for our lab was finally installed in 1972, together with other indispensable instrumentation like gas- and liquid-chromatographs and general laboratory equipment. The lab startup was heavily delayed by politics and red tape, to which I had had no previous experience and exposure, while in the United States. For instance, in July that year, a coordination protocol was signed between the Autonomous University of Barcelona and the Spanish Research Council (CSIC) officially creating the research division of the IBF as a coordinated center between the two institutions. More paperwork but no real advantages for its research teams! Nevertheless, we started working in earnest and our first MS publication did appear in 1974 (Gelpí et al., 1974). In April that year, I received a letter, that I always have kept, from Al Burlingame in Berkeley offering me to come back and join his clinical biomedical area. If accepted my move back to Berkeley should have taken place at the latest in the fall of 1974 because, per his words, they needed "someone of my talents, drive, and organizational capabilities." These were indeed flattering words, and though I was really tempted, I could not find the drive and courage to leave behind all the students that had deposited their confidence in me and dismiss everything I had so painstakingly organized in the first 3 years in Barcelona and which was showing signs of being somewhat successful. I guess that I figured I had invested too much effort and determination to leave it so abruptly. In retrospect, this decision was possibly the most ethical but maybe not the most productive one. To date, I am still not sure it was the best for me and my family. At this point, I should explain that I was trying to replicate in my lab what I had learned in the United States. In other words, as a young and inexperienced Ph.D. candidate you struggle to get accepted by an appropriate theses' director who, along the lines of his/her interest, will guide you to develop your research work for your successful graduation. Once graduated, you are free to look for a postdoctoral position in an institution different from where you earned your Ph.D. There, depending on the place and research group you selected, you can venture into new areas of research. After that, you are totally on your own to find a suitable place to develop your own research and open new lines adjusted to your own interests. As I recall, when I graduated from the University of Houston, I was told that the rules there even prevented me from reapplying in the future for a new position in that University. I will try to explain below why this normal course of events for a young Ph.D. did not happen in Barcelona and where this led me. As indicated above, within the first 2–4 years in Barcelona, eight young students started to work in my laboratory towards their respective doctoral degrees and I laid out to them my plans to open two main research lines, one related to the neurochemistry of biogenic amines as neurotransmitters and their role in neurological diseases, and later, as the first one was taking hold, another on the physiological role of prostaglandins as inflammatory mediators in various clinical conditions. Both were new and challenging research fields. In both cases, I relied on the development of MS techniques, mostly coupled to gas- and liquid chromatography for the detailed study of these metabolites and their physiological roles as biomarkers of human disease. In my thesis, I had developed methods to detect and identify biomarkers of terrestrial and extraterrestrial origin, but after my return from the states, it was evident that it would have been foolish to continue my earlier work on the development of MS and its applications to geochemistry, the analysis of extraterrestrial samples, and the origin of life in general. I simply would not have been able to locally raise sufficient academic interest and financial support. Furthermore, as a recently graduated Ph.D. and because it was only natural, I needed to get away from the research of my former adviser and establish my own research lines. Therefore, I decided to organize a laboratory specialized in biomedical MS in Barcelona. In fact, the dream of becoming a biochemist and running someday my own research unit on the improvement of human health had been my driving force to emigrate to the United States in the first place. For this task to take hold I was forced to secure in Spain the help and complicity of relevant professionals at the University, the Ministry of Education, and private commercial firms. All of it was hard work made especially complicated by the precarious economical support that, despite my efforts, these students were getting from the university or the CSIC. The situation brought back to mind the contrast with my laboratory assistantship in Houston. This position carried with it a monthly salary of $220. Not much by US standards at that time but enough to rent a furnished apartment, get married, and barely make it to the end of the month. Because my standing and productivity at Oró's department were improving, this salary was soon supplemented with NASA grants and/or contracts so that I was able to quit lab teaching duties and concentrate on my Ph.D. research. My students in Barcelona were working as hard as I had done at the Universities of Houston and California and in my view, they were certainly entitled to more generous support. However, because of the way things were then in Spain, and unfortunately, not much has changed to date, most of my time was consumed fighting the rules and prevailing bureaucracy to secure grants and contracts for doctoral and postdoctoral students. In fact, it was relatively easier to get funds for instrumentation rather than for personnel. I need to admit that, during my first years in Spain, I took advantage of my glamorous role as a reclaimed promising scientist and submitted myself to the ordeal of radio, TV, and press interviews. Being part of a reversed "Brain Drain," as it was referred to, carried with it an aura of success. My hope and expectations were that this privileged status could somehow counteract the gridlock of bureaucracy and thus accelerate the assignment of funds for personnel and upgrade the very basic and indispensable instrumentation. This glamour, however, did not last long, and we were forced to make do with what we had. In this regard, I remember with pride the research we did ourselves to upgrade our Hitachi MS. The RMU-6H was not equipped for selected ion monitoring (SIM) and it also lacked a suitable interface for GC-MS work, which was an absolute must for our research projects. In Berkeley or in Houston, I would most likely have had new instrumentation installed for this purpose, but not in Barcelona. So, with outside technical help, we built a homemade four-channel SIM unit that was coupled to the Hitachi MS and we also coupled to it a single-stage gold jet separator for GC-MS. This study was published in Analytical Chemistry in 1977 (Artigas et al., 1977) and proved crucial for our continuing work in the laboratory. Unfortunately, at that time, no computerized control and data handling facilities were available and we defined the mass scale of the spectra aided by column bleed, as mentioned above. Also, the detailed study of molecular fragmentation was facilitated by a very handy mechanical aid. That gadget was the slide rule! (Figure 7). With it, we could easily identify the path of the metastable mass transitions which were very informative about molecular decompositions of ions in transit from the ion source to the magnetic field. Despite all the difficulties, after our first publication in 1974 and up to 1980, we managed to publish 28 papers (selected refs: Artigas & Gelpí, 1979; Artigas et al., 1977; Gelpí et al., 1974; Roselló et al., 1981; Tusell & Gelpí, 1980) on the study of the metabolic pathways and roles of neurotransmitters and prostaglandins. By 1980, all my initial eight students had graduated and obtained their doctoral degrees where they used various liquid chromatographic and GC-MS approaches for their experimental work. Accordingly, I thought they would ask me for recommendations for postdoctoral work within Spain or abroad, however, here is where things started to diverge from all I had learned in the United States! If these young predocs would have done as expected to gain their independence from my direct predoctoral supervision and had settled themselves elsewhere, then I would have been free to admit new students into the lab. Academically, you renew your research team with every graduation. That's what I had thought would happen, however, it simply was not possible because neither the academic nor the business environment was ready to absorb them. Furthermore, the tremendous personal and household mobility I had witnessed in the United States was unheard of at the time in Spain. Things have changed now but then people as a rule studied, married, and got to work in the city or area where they had been born. So, except for three of them, who went elsewhere, the rest stayed and, rightly so, demanded a proper professional stabilization of their postdoctoral situation. After the high dedication and effort shown on their part, I took it upon myself to get them stable tenured positions within the group, and in one way or another, I managed to provide working place stability by ensuring the necessary openings for stable positions. I remember that this task was extremely difficult but that was the least of the problems. Even before gaining access to tenured positions, they soon started to show the normal behavior to be expected from a young postdoc. In other words, they started to claim their independence and bring about their own ideas for new research projects. I had learned that I could not treat them as predocs or qualified lab assistants, as they were naturally looking for their own independent ways. At that time in the early 80s, mostly because of my earlier GC-MS work in the United States and the biomedically oriented publications we had managed to produce from our lab in Barcelona, I was a recognized mass spectrometrist in Spain and abroad. In line with this status, I had been invited to attend the 1982 Vienna International Mass Spectrometry Conference as a member of its Scientific Committee, and in 1983 I was invited to join the Editorial Board of the Biomedical Mass Spectrometry journal. So, I guess that if my students were looking for real academic independence, then they needed to find new areas and accordingly, they slowly started to reorient their interests to projects away from MS. As I see it, there were various factors that explained this academic anomaly, all of them arose from the precarious situation of science that prevailed in Spain. On the one hand, and quite naturally, my recently graduated students felt pressured to build up their own names and own individual reputations and this attitude somehow collided with my personal plans. On the other hand, we were stuck with the old RMU-6H MS, installed in 1972, which we had retrofitted with