September 2016 Volume 29: Supplement
The Chromatographic Society Diamond Jubilee Supplement 1956–2016
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www.restek.com The Chromatographic Society Diamond Jubilee Supplement 1956–2016
6 ChromSoc: The First 60 Years Paul Ferguson An introduction to The Chromatographic Society’s 60th Anniversary Supplement.
8 The Changing Face of The Chromatographic Society and Chromatography in a Changing World Chris Bevan, the longest serving President of The Chromatographic Society in recent times, is interviewed by another former President of The Chromatographic Society, John Lough, on the evolution of The Chromatographic Society and chromatography in a changing world.
13 Progress in LC for Complex Biological Samples — Smaller, Better, Faster Robert S. Plumb and Ian D. Wilson Ian Wilson was a principal speaker at The Chromatographic Society’s Diamond Jubilee meeting on “Advances in Microcolumn and Related Separation Technologies”, which took place in London on 22 March this year. In this article Ian and co-author Rob Plumb elaborate on the theme of the meeting.
17 Past Contributions and Future Visions A popular feature of The Chromatographic Society’s Golden Jubilee publication was an article by Waters describing their heritage and setting out their contributions to the development of chromatography. In this special feature, some of the companies who are regular exhibitors at Society events answer questions on the current and future status of chromatography.
25 Meeting Review: Advances in Microcolumn and Related Separation Technologies Paul Ferguson A review of the first Chromatographic Society meeting in 2016.
26 ChromSoc Activities: The Reid International Bioanalytical Forum Tony Edge, Ray Briggs, and Derek Stevenson This article describes how the Reid International Bioanalytical Forum was initiated and developed over the past 40 years and how its unique philosophy came about.
28 High Impact Chromatographic Developments of the Past 60 Years W. John Lough from the University of Sunderland, UK, in conversation with (old) friends from academia and industry to celebrate 60 years of advances in chromatography.
34 ChromSoc Activities: Medals Chris Bevan and W. John Lough A recollection of The Chromatographic Society’s medals and medallists over the years.
35 Impact of Chromatography on Life and Society Tony Edge Tony Edge, The Chromatographic Society Vice-President, discusses just how much chromatography underpins everyday life.
39 ChromSoc Activities: Academic Support Karen Rome A look at some of the opportunities ChromSoc currently offers.
42 The Future Revisited Peter Myers
Photo Credit: Volker Gugath / EyeEm/Getty Images Peter Myers ponders the future for separation scientists.
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Paul Ferguson, President, The Chromatographic Society, UK.
An introduction to The Chromatographic Society’s 60th Anniversary supplement.
impact of separation science not just in providing you with insight into The industry, but its relevance to everyday Chromatographic Society in a similar life and to ensuring the quality of format to our 50th golden anniversary everything that we consume, that we booklet, which we produced in 2006 use, and even the quality of the air (and may still be viewed on our website we breathe. It is the most important at www.chromsoc.com). As we will branch of analytical chemistry discuss later, we are also looking for and in itself should be celebrated. your feedback regarding what you feel Separation science has developed are the most important developments in so many ways over the lifetime of in separation science over the last 60 the Society — encompassing liquid years (we’ll be sharing some of these chromatography (LC) and its numerous thoughts in this publication) through a forms, electrophoresis and its variants, short on-line survey and will share the supercritical fluid chromatography results early next year. (SFC), miniaturization, and hyphenation We would also like to thank the — not only from a practical companies who have sponsored this supplement. We are fortunate As President of The that the Society has close links with Chromatographic Society, many chromatographic instrument Paul Ferguson, President of The and consumable manufacturers Chromatographic Society (ChromSoc) I hope you enjoy this who support our many activities. As special supplement. We’d you will read in this publication, The It gives me great pleasure to be able be delighted to receive Chromatographic Society is at its core to introduce this special supplement a meetings organization. We organize produced in collaboration with your feedback regarding at least four scientific meetings each LCGC Europe, which is dedicated it, and hopefully it will year in the UK and vendor sponsorship to celebrating the 60th (Diamond) encourage you to join us, of these meetings is incredibly anniversary of The Chromatographic important in allowing us to hold these Society (ChromSoc). Our Society has attend our meetings, and events, but also to support our altruistic its roots in a meeting in London in help support our activities activities. Their sponsorship of this late May 1956 organized by the late in the future. magazine is as equally important Denis Desty to discuss the rapidly as supporting our events because it growing field of gas chromatography perspective, but also from a theoretical provides a platform to showcase our (GC). For those of you reading this understanding too. activities and helps us in our goal of publication and have not heard To help us inform you more about supporting chromatographers. of us, we are a charitable society the Society and various aspects of My final thanks go to John Lough run by volunteers dedicated to separation science, we are delighted of The Chromatographic Society, who “supporting chromatography and to be collaborating with LCGC Europe has been the driving force for this chromatographers”. We do this in many in producing this supplement. LCGC is publication. Without his significant ways — primarily by holding scientific a very important publication in the field efforts, you would not be reading this meetings in the UK but also supporting of separation science and has a huge today. international chromatography meetings, readership that seamlessly connects As President of The through awarding our internationally scientists at the bench with the latest Chromatographic Society, I hope recognized Jubilee and Martin Medals in practical and cutting-edge research. you enjoy this special supplement. (which you may have seen presented We very much hope to work with LCGC, We’d be delighted to receive your at the HPLC or ISC meetings this not only on this publication, but also in feedback regarding it, and hopefully year) and awarding travel bursaries to the future too. it will encourage you to join us, attend enable students and chromatographers We hope this celebratory magazine our meetings, and help support our based in industry to attend national and containing historical articles, activities in the future. international separation science events. interviews, and technical articles If you are reading this publication, reflects some of the fundamental Paul Ferguson is the President of The you are undoubtedly aware of the values of LCGC while also Chromatographic Society.
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©2016 Waters Corporation. Waters, The Science of What’s Possible, and ACQUITY are registered trademarks of Waters Corporation. Arc is a trademark of Waters Corporation. The Changing Face of The Chromatographic Society and Chromatography in a Changing World
Chris Bevan, the longest serving President of The Chromatographic Society in recent times, is interviewed by another former President of The Chromatographic Society, John Lough, on the evolution of The Chromatographic Society and chromatography in a changing world.
John Lough: Going back to when pharmaceutical company venues the surpluses, although high, were you first became President in 2000, with the commensurate increase in too infrequent to deliver a regular and it was actually quite an unusual rates for exhibiting involved. Was sufficient income stream to meet our event. For only the second time this some kind of grand master expenditure. Furthermore, we felt that in two instances from 1990 to plan or were you just having to more countries should be allowed and 2016 it came down to a vote. In respond very quickly to changing encouraged to host these meetings keeping with the theme of this circumstances? More importantly, and therefore the ChromSoc’s interview, the issues were around what was the exact nature of these proportion of the surplus would be the external context. In challenging circumstances? less in the future. The Society has circumstances, it was down to CB: Surprisingly the challenge was always had support from its volunteer whether The Chromatographic not just from the difficult prevailing committee members and from various Society could continue to thrive as external financial circumstances. companies who allowed us to use their an independent group or whether I have mentioned our “outdated premises for meetings for free or low it was necessary to cooperate financial model”. We were, if you rentals. We ran an office for many years extensively or actually merge with like, the author of our own potential with two secretaries and for these we other bodies operating in the same downfall by our insistence in doing were paying full commercial rates. or overlapping areas. Looking what was right for chromatography. I reviewed the charges we were back now, do you still believe that The Society was one of the three asking from companies to exhibit retaining the Society and its name founder members of the International their products at our meetings and was the correct thing to do to meet Symposium on Chromatography realized that they were insufficient to the challenging circumstances at (ISC) series of symposia along with meet our expenditure. Closing offices that time and has that decision the French and German separation to save money is the emotionally easy served us well since that time? societies. The financial surpluses from bit, but making old friends and loyal Chris Bevan: I most certainly these symposia provided the Society colleagues’ jobs redundant is another do believe it was right to retain with most of its funds. Unfortunately, matter altogether. The way I saw it was the Chromatographic Society’s well-established name and to continue to promote its mission as Figure 1: Chris Bevan, former President of The Chromatographic Society (2001–2006). an independent scientific charity representing the interests of separation scientists. The problem was our outdated financial model, not our good reputation, and performance in delivering symposia and discussion meetings on chromatography and allied techniques. To dilute our impact by merging us with other groups would have weakened us and I suspect we would have disappeared.
JL: You went on to transform the Society’s finances (a) by reorganizing our management with the concomitant closure of our office, and (b) moving more towards symposia held in major
8 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Chromatography in a Changing World
that if we didn’t act quickly then we Figure 2: Chris Bevan changed the way that Society meetings were run, in particular wouldn’t have enough money to pay a with respect to the use of Big Pharma venues for meetings. The peak of this trend was fair redundancy payment to our loyal in ChromSoc’s Golden Jubilee year when Chris was the mastermind behind the Triad staff so the decision was inevitable if series of meetings, which were held through the year at AstraZeneca (Loughborough), Pfizer (Sandwich), and GSK (Stevenage). Here we see an image of the impressive Pfizer the society was to continue. (Sandwich) site that was used to promote the 2006 Spring Symposium. JL: You have held an important position in a major multinational pharmaceutical company. The Society had strong links with UK pharmaceutical companies and so it was fitting that the Society celebrated its Golden Anniversary in 2006 with your Triad series of Big Pharma meetings. Times have changed. The Triad could not be repeated today. You are ideally placed to comment: Have you any observations to make on the changes in the pharmaceutical industry in the last 10 years? Have these changes had an effect on “what chromatographers do”? CB: I have witnessed the global expansion and subsequent contraction of the pharmaceutical industry over the past 40 years — both from an international and a UK perspective — having been employed by Hoechst,
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www.chromsoc.com 9 Chromatography in a Changing World
Glaxo, GlaxoWellcome, and GSK. The scientific innovations and their Figure 3: Chris Bevan with Chromatographic Society associates, Ted Adlard and Steve Rumblelow, alongside a regular distinguished speaker at ChromSoc meetings, realizations coupled with the wide Pat Sandra, who is with his wife Martina. The photo was taken at the November 2013 application of ingenious technologies meeting on Advances in Natural Product Analysis: Separating The Wheat From The have impressed me. The changing Chaff at the Syngenta Research Laboratories in Bracknell, Berkshire, UK. markets and the lack of drug-sensitive targets and diseases for which there are no animal models have challenged the industry. Chromatographers used to be an elite group within analytical and process science departments, but nowadays they are dispersed and have become nonspecialists. Much of the analysis that used to be performed manually is now fully automated and run in a preprogrammed manner. The oddball situation is often ignored if the result doesn’t quite fit into the automated systems agenda. Chromatographic scientists now must have a broader appreciation and understanding of mass spectrometry delivering separation science Europeans, while the number of (MS), nuclear magnetic resonance activities in Europe, with all the UK delegates attending European (NMR), and the world of the disparate groupings failing to conferences was dwindling. The large-molecular-weight biomolecules, come together. Looking back, Society’s horizons must be determined rather than the small drug molecules. notwithstanding the point that by and reflect the interests of its The advances in computational ChromSoc committee members members and attendees and these molecular modelling have also had an do not have infinite time available seem to be UK-centric at present. impact in separation sciences and in to them, do you think that was a European members and non-members physicochemical property prediction moment when ChromSoc could have have always been welcome to come to and measurement. grasped the nettle and widened its our national and international meetings It is quite unusual to find an isolated horizons to become a Europe-wide and have done so in considerable specialist chromatographer in big Chromatographic Society? numbers over the years. pharma anymore and multidisciplinary CB: As the Society’s Honorary analytical teams have been disbanded Secretary and later as its President I JL: Your current role involves because of economies and a was very much in favour of maintaining being one of the principal meeting reluctance to risk future venture capital the Society’s connections with societies organizers. Elsewhere in this on longer-term goals. Late failures abroad and met many scientists who supplement, there are several other to deliver new profitable drugs in a were equally passionate to cooperate individuals looking ahead to the reasonable time is making even large internationally. A great deal of sterling future of chromatography. So, the companies somewhat risk-averse. work was undertaken by the Germans, question here is simple — what are It is important that the young French, Austrians, Belgians, Italians, the areas of chromatography that multidisciplinary chromatographers Spanish, Scandinavians, Polish, and are currently exciting you and which of today still strive to maintain the the Hungarians to establish the legal will feature in your forthcoming excellent team spirit and inventive framework and financial foundations of meetings? cooperation that has always been EuSSS. I went to a number of excellent CB: I see the further development the hallmark of the UK’s pharma R&D conferences and committee meetings and application of supercritical fluids scientific fraternity. that were run by these pioneering to chromatography coming back into EuSSS scientists. However, it became vogue to enable purification JL: Again, looking outside of the apparent that most of the European and isolation of biochemicals and Society, there are other societies separation science and chemical natural products. I am told that working on the development of societies had their primary focus it is more selective, eco-friendly, chromatography. I am thinking on running symposia in their own and energy efficient than using here mainly about Europe. You countries. The emergence of Eastern conventional solvents. On the were noted in the early 2010s for European countries also affected analytical scale I see progress in the your sterling efforts in trying to the financial model for running a further miniaturization and automation modernize the European Society Europe-wide society, encountering of separations science, particularly for for Separation Science (EuSSS). many similar issues to those facing the separation of complex biological However, you eventually came to the Eurozone countries today. Our molecules and those related to the the conclusion that EuSSS was UK-based Chromsoc symposia were understanding of the human genome ineffective at coordinating and running well and attracted many and its complex mechanisms. The
10 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Chromatography in a Changing World
techniques are essential prerequisites Figure 4: 2016 and Chris Bevan is still organizing successful meetings for The for success. Chromatographic Society. The next meeting will be on Automation & Data Handling for Diverse Separation Science Applications at the Syngenta Research Laboratories in JL: You have seen and heard a Bracknell, Berkshire, UK, on 25 October 2016. lot, so, to go back to the meetings you have staged in the past 60 (sorry, 25–30!) years and chromatographers you have met at Chromatographic Society and international meetings, are there any memories you’d like to share with us? Who were your favourite speakers? Who impressed you most as a chromatographer? CB: Following a PhD in neutron activation analysis and chemical oceanography, I started applying and developing liquid chromatography (LC) methods for blood drug analysis with Hoechst Pharmaceutical Research laboratories back in 1976 using early Waters high performance LC (HPLC) equipment. I was curious to meet and get to know Jim Waters and at the ChromSoc 50th anniversary “Triad” meeting in 2006 at GSK, I had the honour of presenting him with the biggest current challenge for analytical fields, and the coupling of Martin Medal. Talking of Archer Martin, chemistry is from the complex chromatographic and electrophoretic I heard his last lecture because he biochemical and biological-related techniques with structural elucidation was our invitee plenary speaker at the
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www.chromsoc.com 11 Chromatography in a Changing World
London ISC symposium. Tragically, the to catalyze these discourses. I hope for my PhD. Gamma ray photo peak next time I saw Archer Martin was on that the society will continue to foster areas and background subtraction a BBC TV programme on Alzheimer’s such opportunities for all scientists were calculated and processed on a with his daughter. His Nobel to ensure that chromatography and large mainframe computer at Harwell’s Laureate-winning brain had been made electrophoresis are recognized by Rutherford labs ATLAS computing dysfunctional by plaques; surely a the public as backroom tools in the centre and at the University of London’s challenge for future chromatographers analysts’ armoury, so essential to many IBM370, employing a program called to make sure they make an important companies. SAMPO written by Routti (1). Data was contribution to the underpinning Should any one reading this have recorded onto punched paper tape technological advances needed to an aspiration to become a Society and punched cards from the 4096 elucidate the mechanisms of this evil committee member don’t hesitate — it multichannel analyzer gamma ray and disabling disease. will enrich your social and scientific spectrometer. The instrument produced Discussions in the bar and at dinners life and you will meet a great bunch of massive amounts of data very quickly and social events at symposia are the people and their families. and had I not had the computers and places where one has an opportunity to programs I would still be there now get to know other scientists personally JL: Thanks, Chris. That’s a very with my slide rule and calculator. I and gain insights into their work. I uplifting thought with which to could have shaved at least six months was particularly delighted to see that finish. Thank you very much for your from my PhD if I had had access to a my freeze-thaw switching invention time and, by way of thanks, we will modern PC. had found an important application give you a final bonus question! Nowadays, no one considers it in the analysis of the human genome Several others have been given the unusual to deconvolute overlapping as a result of a casual conversation opportunity through other articles in peaks, baseline subtract, or derive at York University HPCE course with this publication to give their views linear and non-linear least squares Ed Yeung. Ed had mentioned that on what have been the important regressions for calibration: they use he had problems switching flows developments in chromatography many sophisticated statistical functions and derivatizing analyte peaks in his over the past 60 years and we hear to measure the quality of their data. multiple (96) capillary electrophoretic from you again. However, for you, So, the most historical development systems and needed a reliable, what has been THE major step must be computational power and such very low volume valve. I had been forward? easy access to it, with chromatographs developing a suitable method for CB: Today this may seem really becoming perhaps more intelligent achieving this and he decided to apply obvious to our readers, but back than their operators before too long! it in his laboratory. This sort of instance in the 1960s when I was doing my is what makes innovative science so BSc degree in analytical chemistry Reference rewarding. it would not have been predicted (1) J.T. Routti, SAMPO, A Fortran IV Program for Computer Analysis of Gamma Spectra Nearly 50 years in analytical by the average analyst. In the late from Ge(Li) Detectors and for other chemistry and associated sciences 1960s most analytical chemistry was Spectra with Peaks (UCRL-19452, UC-34 have left me with a perspective that performed manually and without PHYSICS. T10-4500, Lawrence Radiation I believe is unique. The scales of sophisticated instrumentation. Burettes Laboratory, University of California, Berkeley, USA, 1969). chromatography I have worked on and pipettes for wet volumetric have been from the ultraminiaturized analysis and balances for gravimetric Chris Bevan is a former Group 50-μm CE capillaries to the 6-inch analysis coupled with quantitative Leader of the physicochemical diameter large-scale prep columns, measurements obtained from characterization team at GSK all performing separations: a scaling manually transcribed readings from and former President of The factor range of over 3 log orders. spectrometers were commonplace. Chromatographic Society. He is now Determining nanogram per millilitre Statistical measurements derived from retired and works as Chromsoc’s concentrations in trace analysis to tens many thousands of determinations were Events Coordinator. of grams separated out preparatively unusual and quite tedious to perform, John Lough is a Reader in — all-in-a-day’s (or a career’s) work for even with the help of a mechanical Pharmaceutical Analysis in the the practising separations scientist. calculation machine (the FACITs and Sunderland Pharmacy School at I have thoroughly enjoyed my career Brunsvigas were my mechanical the University of Sunderland, in in science and met many excellent GameBoy toys at that time). Sunderland, UK, and a member scientists in many different countries. The ubiquitous peak area of the Executive Committee of The I admire and enjoy the company of measurement for quantitative Chromatographic Society. those people who are willing to share calibrations was done by printing out their thoughts and make real attempts on a chart recorder and cutting out the at explaining their discoveries to others peaks and weighing them — I kid you The next meeting will be on who may not be from their discipline not! Automation & Data Handling for or understand the specialized terms I was very fortunate to start my Diverse Separation Science of their language. Interdisciplinary analytical career immersed in the world Applications at the Syngenta Research discussions are fundamental to the of nuclear physics. Nuclear physicists Laboratories in Bracknell, Berkshire, solving of most of today’s problems had access to the best tools for rapid UK, on 25 October 2016. and a convivial atmosphere is vital calculations and I used them widely
12 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Progress in LC for Complex Biological Samples — Smaller, Better, Faster
Robert S. Plumb and Ian D. Wilson, Division of Computational and Systems Medicine, Department of Surgery and Cancer, Imperial College London, London, UK.
Ian Wilson was a principal speaker at The Chromatographic Society’s Diamond Jubilee meeting on “Advances in Microcolumn and Related Separation Technologies”, which took place in London on 22 March this year. In this article Ian and co-author Rob Plumb elaborate on the theme of the meeting.
Chromatography has always had from 5 μm to 3.5 μm starting from 3.0-mm i.d column-based assay to a a major role in the separation the early to mid-1980s were major 2.1-mm i.d format proved surprisingly of complex mixtures and in improvements, but were not achieved challenging (1), in no small matter the last half-century there has easily. The improved performance related to the dead volume and been remarkable progress in of these phases also resulted in a dispersion in the chromatography the tools available to the liquid reduction in column length from system and connecting tubing. chromatographer for routine 25 cm to 10 cm or 15 cm for most However, once the difficulties applications in separations and routine applications. For those of associated with this further reduction analysis. These are now incomparably us working in application-focused in column internal diameter had better than those available in the industrial laboratories, attempts at been overcome, it did provide gains 1970s when high performance miniaturization for routine analysis in reduced sample and solvent liquid chromatography (HPLC) for a long time consisted of reducing consumption together with increased began to make serious inroads into column internal diameters from sensitivity and better compatibility general analytical laboratories. The 4.6 mm to 3.0 mm, which whilst with mass spectrometry (MS). Further early relatively primitive systems relatively easy and, no doubt, useful reductions in column internal diameter (25 cm × 4.6-mm i.d columns packed did not exactly represent a paradigm to 1 mm, however, did not readily with 5–10-μm packing material, shift. However, moving a pre-existing yield further gains in sensitivity and sometimes also very primitive pumps and detectors, coupled to Figure 1: High resolution reversed-phase UHPLC–MS analysis of human chart recorders for data capture) urine. Adapted and reproduced with permission from Analytical Chemistry were a world away from the modern 78(20), Robert.S. Plumb, Paul Rainville, Brian W. Smith, Kelly A. Johnson, Jose instruments currently found in the Castro-Perez, Ian D. Wilson, and Jeremy K. Nicholson, Generation of Ultrahigh laboratory. Compared to the available Peak Capacity LC Separations via Elevated Temperatures and High Linear gas chromatography (GC) systems Mobile-Phase Velocities, 7278–7283 (2016)¶American Chemical Society. they were expensive (for many years the “HP” in HPLC was presumed by 100 some to mean “high priced” rather than high performance), unreliable, and incapable of achieving the performance of GC for complex mixture analysis. However, the potential was obvious, and despite the clear problems, the major advantages that HPLC offered in terms of its ability to deal with % (relatively) polar and involatile
analytes made it extremely attractive Intensity to separations scientists faced with the problems of analyzing biological samples. The intervening 50 years since the introduction of HPLC has been a story of steady, if irregular, progress in both stationary phases and instrumentation. So the methods 0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 for the routine production of spherical silica particles, with a narrow size Time (min) range, and reduction in particle sizes www.chromsoc.com 13 Complex Biological Analysis
this was probably a result of the difficulty in packing them efficiently. Figure 2: Base peak intensity (BPI) mass chromatograms with +ve ESI detection In parallel with these modest human urine (a) 2.1 mm i.d. and (b) 1 mm i.d. 10 cm UHPLC columns. The solvent advances in LC there were major gradient profile was the same for both separations with the flow rate scaled from 0.6 mL/min to 0.14 mL/min. The inset shows extracted ion chromatograms for gains in sensitivity, specificity, and kynurenic acid showing increased response for the 1-mm separation. Adapted the ability to identify peaks eluting and reproduced with permission from Journal of Proteome Research 14(6), Nicola from the column because of the Gray, Matthew R. Lewis, Robert S. Plumb, Ian D. Wilson, and Jeremy K. Nicholson, introduction of efficient methods for High-Throughput Microbore UPLC–MS Metabolic Phenotyping of Urine for hyphenating the technique to mass Large-Scale Epidemiology Studies, 2714–2721 (2015)¶American Chemical Society. spectrometers. These advances in detection significantly helped with (a) 2.1 mm i.d. enabling the application of HPLC to 100 ESI+ complex mixture analysis in areas such as proteomics, metabonomics or metabolomics, drug metabolism, % clinical analysis, and environmental analysis, and helped to mask the (a) 2.1 mm i.d. relatively slow progress in the 0 100 B/N. P/P. 151 development of LC itself. However, 1.00 2.00 3.00 4.00 5.00 % (b) 1 mm i.d. 0 there was no disguising the problems 3.00 3.50 4.00 4.50 100 (b) 1 mm i.d. S/N. P/P. 190 associated with LC–MS, the most 100 2.6 x peak area
egregious of which was probably % 2.4 x peak height 5 x S/N 0 ion-suppression, resulting from 3.00 3.50 4.00 4.50
co-elution of compounds. For this % reason the next major gain for Intensity Intensity complex mixture analysis came from a further reduction in particle size 0 1.00 2.00 3.00 4.00 5.00 6.00 7.00 from 3.5-μm to sub-2-μm packing Time (min) materials for chromatography and leading to the introduction of ultrahigh-performance LC (UHPLC). it is no longer necessary to sacrifice might be thought that these reduced The availability of sub-2-μm packing an animal for each time point: the sample sizes have less benefit for materials was not enough on its own entire blood concentration versus studies in larger species, including but these materials also required time profile can be obtained from a humans, where obtaining larger the development of solvent delivery single animal. So, whilst detection sample volumes is less problematic. and autosampler systems capable limits for a particular analyte may still But in, for example, epidemiological of providing higher pressures than be 1 pg/mL, it now only requires a studies or biobanking, where “conventional” HPLC systems were 20-μL blood or urine sample to be collections of samples can number capable of. Indeed, ensuring that the obtained for analysis, rather than the hundreds to hundreds of thousands, benefits of separations on UHPLC 100–1000 μL needed in the past (and the storage of large volumes comes columns were not lost required the the analysis is over in less than 5 min at significant economic cost so that, optimization of the whole analytical compared to 10–15 min!). So it is now even here, the benefits of sensitive system if performance was not to possible in theory, and in practice, analysis are very clear. be compromised by extra column to obtain from a single 20-μL blood band broadening. The results of the spot or microsample target analysis The early relatively introduction of UHPLC for complex for a drug, qualitative analysis for mixture analysis were transformational drug metabolites, and a metabolic primitive systems were and this explains why the penetration phenotype of the sort needed for a world away from the of such systems into general routine metabonomic or metabolomic modern instruments use was so rapid. An example of the analysis for detection of biomarkers application of UHPLC–MS for urine of efficacy and toxicity. Indeed, currently found in the analysis is given in Figure 1. with appropriate design of the laboratory. The combination of efficient analysis method, a single “all-in-one” chromatographic separations determination of all of the above can Congruent with the development together with sensitive detection has be done. These new opportunities of UHPLC it is also clear that the also powered another revolution as represent real benefits, including a quality of the 1-mm i.d. microbore they enable significant reductions in possible 10-fold reduction in animals UHPLC columns had much improved sample size. In the case of studies used and reduced costs. The major compared to the microbore HPLC in rodents this has resulted in the benefit, of course, is better science columns that we had tried to use potential for a dramatic reduction in because the resulting PK profiles in the past and was now sufficient animal use, such as in, for example, are from individual animals rather for them to be considered as a pharmacokinetic (PK) studies, where than being a composite of several. It replacement for 2.1-mm i.d columns
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for routine metabolic phenotyping. + The resulting method (2), which Figure 3: A chromatographic trace obtained from the UHPLC–MS ( ve ESI) analysis of a rat urine sample using the RAMMP method. Adapted and reproduced with permission required minimal alteration to a from Analytical Chemistry 88, Nicola Gray, Kyrillos Adesina-Georgiadis, Elena standard UHPLC system, was able to Chekmeneva, Robert S. Plumb, Ian D. Wilson, and Jeremy K. Nicholson, Development demonstrate equivalent or superior of a Rapid Microbore Metabolic Profiling Ultraperformance Liquid Chromatography−Mass performance in terms of peak Spectrometry Approach for High-Throughput Phenotyping Studies, 5742–5751 (2016)¶ capacity, sensitivity, and robustness American Chemical Society. to the conventional method, giving (on average) a threefold increase 100 in sensitivity over the conventional 2.1-mm method and showing no decline in performance over the course of 1000 analyses of human urine samples. Examples of the separation of a urine sample at both column geometries are shown in Figure 2. Thus, miniaturizing our % UHPLC–MS method for metabolic Intensity phenotyping gave both economic and environmental benefits from a reduction of 75% in solvent consumption and similar or improved analytical performance compared to 0 conventional UHPLC–MS. 0.00 1.00 2.00 We believe that this example demonstrates that it should now be Time (min) possible to implement microbore methods for routine, large sample number assays with confidence. solvent into the source, or we would where complex samples must be However, throughput remains a have had to split the flow. These characterized and their components potential problem in situations where changes reduced peak capacity identified. For the future, developments there are large numbers of samples from 150 for the 2.1 mm × 10.0 cm in robust capillary LC separations and limited analytical capacity column method to 50 for the RAMMP and the more widespread use of ion methods for speeding up the analysis approach, with a corresponding mobility spectrometry provide further are required. An obvious response reduction in the number of features opportunities for enhancing our to increasing throughput is to reduce detected (from 19,000 to ca. 6000). abilities to perform rapid, routine, and the run time but, for complex mixture However, despite reduced peak complex mixture analysis analysis, this does pose a problem capacity and feature detection, the because, within limits, the longer the RAMMP method still achieved similar References time allowed for the separation the levels of group discrimination as (1) K.A. Oudhoff, T. Sangster, E. Thomas, and I.D. Wilson, J. Chromatogr. B 832, more features that will be detected. conventional UHPLC–MS. If 191–196 (2006). Nevertheless, in early studies we had the compromise between speed (2) N. Gray, M.R. Lewis. R.S. Plumb, I.D. demonstrated that even with a short and metabolome coverage is Wilson, and J.K. Nicholson, J. Proteome analysis time sufficient information accepted, then the RAMMP approach Res. 14, 2714–2721 (2015). (3) R.S. Plumb, J.H. Granger, C.L. Stumpf, was still obtained from rat urine provides a robust and sensitive K.A. Johnson, B.W. Smith, S. Gaulitz, samples to enable them to provide method that delivers high-throughput I.D. Wilson, and J. Castro-Perez, Analyst useful metabolic phenotypes (3). metabonomic analysis of complex 130, 844–849 (2005). (4) N. Gray, K. Adesina-Georgiadis, E. We therefore combined a reduced mixtures such as urine combined Chekmeneva, R.S. Plumb, I.D. Wilson, analysis time with 1-mm microbore with a fivefold reduction in analysis and J.K. Nicholson, Anal. Chem. 88, UHPLC in a high-throughput “rapid time compared with the conventional 5742–5751 (2016). microbore metabolic profiling” UHPLC–MS method. And of course, if (RAMMP) approach (4) by scaling the appropriate, samples can always be Ian D. Wilson is a Professor of Drug 2.1 mm × 10.0 cm column method interrogated in more detail using more Metabolism and Molecular Toxicology described above with its 12.5-min run comprehensive separations in the Department of Surgery and time down to 1 mm × 5.0 cm columns (or different modes of Cancer at Imperial College, in so as to provide an analysis time of chromatography such as hydrophilic London, UK, and a Chromatographic 2.5 min/sample (Figure 3). In addition, interaction liquid chromatography Society Martin and Jubilee Medallist. we increased the linear velocity by [HILIC]). Robert S. Plumb is the Director of a factor of 4. This was only possible The current suite of chromatographic Metabolic Phenotyping for the Waters using a 1-mm column, because if options available for the analysis of Corporation and Visiting Professor at we had used a 2.1-mm format we complex mixtures are enabling rapid the same department as Ian Wilson at would have been putting far too much progress in a wide range of fields Imperial College, in London, UK.
16 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Past Contributions and Future Visions
A popular feature of The Chromatographic Society’s Golden Jubilee publication was an article by Waters describing their heritage and setting out their contributions to the development of chromatography. In this special feature, some of the companies who are regular exhibitors at Society events answer questions on the current and future status of chromatography.
wider range of scientists. The initial exceed those of more conventional separation scientist was a highly technology. The academic world is trained academic who had the leading the way here, but they are Agilent Technologies ability to do front-line maintenance, being supported by manufacturers column manufacture, and complex such as Agilent, who are keen to Overall, what have been the data interpretation. The greater better understand the advantages most important developments in use of computer technology has of this approach to improving the chromatography over the past 60 vastly simplified the interface analysis of complicated sample sets. years (other than those made by with the instrumentation and the The application of this technology to your company)? advancements in the robustness of real-world analysis and in particular The developments in capillary GC the hardware have ensured that the the application to complicated and HPLC have seen some of the accessibility of the technology has protein samples has a very exciting most important developments. reached a much wider audience. future. In order to get the best of this Reference could be made to This does present challenges in technology, it is important to have the development of the particle ensuring that the technology is identified the types of stationary technology, with initially fully porous robust enough for customers. phases that need to be employed material and then the development to drive the best separation and the of superficially porous materials How does your company see technology required to support this. for HPLC. However, the biggest the current state-of-the-art of developments really have been the chromatography and, in this Has your company thought about coupling of separation technology context, what are your company where chromatography might be with detection technology — in doing to address unmet needs? in 10 years’ time? If so, how might particular the coupling of mass Agilent prides itself on the future look? spectrometry. Mass spectrometry understanding what the customer The future of chromatography is coupled to separation devices has really wants, and delivering solutions very difficult to predict, although revolutionized many industries in to support this. Customers have real if the last 10 years are anything to terms of the detection limits that samples to analyze and this can go by there have not been so many have been achieved, and also for be forgotten in the drive to provide significant changes. The biggest the identification of compounds, new exciting solutions. The sample users of chromatography are still the particularly in the field of protein types that are being analyzed are pharmaceutical industry, however, analysis. becoming more complex though, the focus is changing, moving from and this will drive the separation laboratories based solely in the What has been your company’s technology to greater separation West to a staggering growth in Asia highest impact contribution to power. Improving the separation Pacific countries. The growth of the chromatography over the past through brute force in HPLC, biopharmaceutical industry also has 60 years? Why you think the by simply changing the particle the potential to change the types development had such a high size, has seen improvements in of chromatography that are being impact? the separation performance for used. I think for certain there will be Agilent has been market leaders customers, however, this is not a improvements in the sensitivity of in both HPLC and also GC and so step change and certainly with the detectors, and also a reduction in choosing between these is difficult. developments of biopharmaceuticals the footprint. I think in terms of the The key to the success has very and the incredibly complicated chromatography, the more complex much been based on state-of-the-art mixtures comprising many tens of samples that customers are needing instrumentation that keeps on thousands of individual components, to separate will result in more performing, with industry standards the world of separation science does advanced forms of chromatography such as the Agilent 1200 series need to see a step change. becoming more commonplace, and HPLC Systems and the Agilent 7890 One of the approaches that the use of 2D LC will start to become GC system. is being investigated by Agilent mainstream. The biggest impact that this is the use of multidimensional The world of GC has seen technology has had is in making chromatography, an approach that developments and incremental
Photo credit: Stockbyte/Getty credit: Images Photo the market more accessible to a can provide resolutions that will far changes will continue to happen
www.chromsoc.com 17 Past and Future Visions
over the coming years. Looking was (and still is) the area where be a gold-standard reference back over the previous 10 years, it many of the errors and much of the technique, but we see more and more is a well-established industry with cost associated with GC analysis applications that require a much a large degree of inertia to change, resides. Anatune, together with greater density of data than GC (or driven primarily by the regulators of some other like-minded small LC) can currently deliver. Whichever the industries where the technology companies around the world, has way you look at these applications, is employed. Certainly the made the automation of sample the fundamental problem is that developments in mass spectrometry preparation and introduction a chromatography is slow — it takes will have an effect on the capabilities practical proposition; lowering costs time to generate theoretical plates. of gas chromatography. and improving data quality as a Right now, this is chromatography’s There is other technology that consequence. biggest weakness. At this point I catches the headlines such as don’t see where the improvements SFC and CE, however, these How does your company see needed are going to come from; but technologies, although they have the current state-of-the-art of you never know — there are a lot of their niche, have never really been chromatography and, in this clever people in this business. employed within the mainstream context, what are your company Anatune has always been a analytical laboratories as front-line doing to address unmet needs? dyed-in-the wool GC company, and separation instruments. Inertia has When compared to other for the first time we have to use driven this to a certain extent, but technologies, the truth is that non-chromatographic technologies the technology in both areas has analytical chemistry is slow to (such as selected-ion flow-tube yet to demonstrate a quantum leap change. The drive for better quality [SIFT]-MS) to give us the speed in performance over conventional data is good, but it does mean that of analysis some customers are liquid and gas chromatography. nowadays, the costs of implementing demanding. innovations are considerable. As a — Tony Edge, R&D Manager, result, for established applications — Ray Perkins, Agilent Technologies there is significant resistance to Managing Director, Anatune change unless there are compelling economic reasons to move away from the status quo. This is an observation, not a criticism. For Anatune, 20 years on from when we started, our mission to make Anatune the automation of GC sample preparation and injection the norm is Crawford Scientific Overall, what have been the still work in progress. most important developments in We pay attention to emerging Overall, what have been the chromatography over the past 60 trends and, currently, issues most important developments in years (other than those made by broadly related to health, safety, chromatography over the past 60 your company)? and environmental considerations years (other than those made by Anatune has been largely involved in are a significant source of the new your company)? GC since its formation and the most applications that we come across; Crawford Scientific has extensive important developments here are: with these emerging applications expertise in column technology the air-bath oven, a genius bit of there is much more scope for and applications, and so our design that enables different heated innovation. focus on the important innovations zones to be linked together without naturally falls with the column cold-spots; the fused silica capillary Has your company thought about technology, which includes the column, which made high-resolution where chromatography might be development of spherical silica with GC a practical proposition; and the in 10 years’ time? If so, how might low surface activity post-bonding low-cost bench-top, quadrupole MS the future look? and high mechanical strength. detector — How did we ever manage As we discussed earlier, there is These have been used for a host of without it? a lot of inertia built into separation applications including the plethora science — so in 10 years time, I of reversed-phase phases that are What has been your company’s think for established applications, available as well as chiral and mixed highest impact contribution to things will look much the same mode ligands. Latterly of course, chromatography over the past as they do now — instrument high efficiency phases have been 60 years? Why you think the (especially mass spectrometer) produced using sub-2-μm particles development had such a high performance will be better and this as well as being adapted to produce impact? will make the automation easier, core–shell morphologies that lend Anatune’s existence flowed from my but it will be evolution rather than higher efficiency without the large belief that insufficient attention was revolution. increases in back pressure. being given to sample preparation For emerging applications things In gas chromatography (GC), the and introduction for GC–MS. This will be different. GC–MS will still development of coated capillary
18 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Past and Future Visions columns was revolutionary and the of knowledge across the whole that we lack high quality, innovative, improvements in efficiency and peak spectrum of analytical techniques and inquisitive people — absolutely capacity propelled the technique and application areas. It is this not. Our role is to ensure that our into widespread use globally. experience, combined with the best clients have the right products to The development of polymeric columns and consumables available, enable them to do the best science substrates for liquid phase which we bring to bear when they can, and where they need separations heralded techniques supplying products and services some assistance to really push such as ion chromatography (IC) to our clients. In recent years we the envelope using new products and size-exclusion chromatography have begun to build our specialist or applications, then our large (SEC), which are so useful in analytical laboratory services with technical team is able to bring their industrial applications. the acquisition of Hall Analytical own wide-ranging experience and Important developments in Laboratories in Manchester, UK, expertise into play. instrumentation include the which allows us to support our Many challenges still remain in the development of (somewhat) clients’ requirements within our own industrial application of analytical affordable high resolution and facilities as well as at the client site. science: high mass accuracy detectors for We are a small, independent, The growth of biopharmaceutics HPLC and GC and of course the chromatography products and has seen rapid change in the way robust interfacing of HPLC with services provider based in the we characterize and measure these mass spectrometric detection via United Kingdom, however, through substances. We continue to work with the atmospheric pressure interface the vision of our Managing Director, clients to improve their capabilities (API) in the early 1980s was Sam Crawford, we have been able for understanding entities such as worthy of the Nobel Prize, which to make a global impact in learning monoclonal antibodies and other the technology earned. Multistage and development within analytical protein-based therapeutic agents. mass spectrometry brought huge science. We have been providing We will continue to assist clients advancements in sensitivity and the classroom and on-site training in to take advantage of the latest introduction of the computing data analytical science for 25 years, column technologies to improve the system (CDS) combined with the however, our global presence has quality and timeliness of their data automatic sampler delivered massive grown through the provision of our and to meet evolving regulatory efficiency savings in data acquisition on-line professional development requirements. and reporting. website CHROMacademy (www. Our experts at Hall Analytical chromacademy.com). This has been Laboratories are specialists in mass What has been your company’s in development for over 15 years, spectrometry and we continue to highest impact contribution to and in the early days, on-line and work with clients to provide advanced chromatography over the past computer-based learning were data and help them to understand 60 years? Why you think the not as well accepted as they are the information available from development had such a high now. However, the continuation spectrometric techniques. impact? of this programme was never in We will also continue to research Our biggest impact has been our question and today we are proud into new and effective ways of ability to remain independent, and to have many tens of thousands delivering the right information at the to offer our clients the best products of members worldwide, who all right time via the right platform to our and services to enable their science. receive much-needed training and clients globally. We have assembled a team of very development from the learning committed sales and technical resources within CHROMacademy. Has your company thought about staff, who work with a vast range of This global presence was in no small where chromatography might be resources to find the right products part made possible through our in 10 years’ time? If so, how might and applications to help solve partnership with LCGC magazine, the future look? our clients’ problems and enable who helped us to market and sell the For the most part, chromatography them to do better science. We can product on a worldwide basis. relies on chemistry to provide then help our clients to implement individual components for detection. these solutions through a range of How does your company see Whilst this remains the case, a consultancy services. the current state-of-the-art of “column” of some sorts will always We are small enough to remain chromatography and, in this be necessary, no matter what form very flexible and responsive to our context, what is your company it takes (miniature, narrow, encased, customers’ needs and have worked doing to address unmet needs? etched onto a chip, etc.). So, we hard to maintain our independence We continue to work with our will continue to support separation whilst continuing to work with the customers to provide knowledge and science through the provision of world’s leading suppliers who trust expertise. the best chemistries to enable our us to deliver a best-in-class service As any science matures, the clients to perform the separations on their behalf. spirit of innovation and necessity of they need. We have served analytical science invention decline in favour of higher This being said, we can see for 31 years and in those years the throughput and generic ways of systems becoming much more company has built a huge amount working. Of course this is not to say integrated, with lower dead volumes www.chromsoc.com 19 Past and Future Visions
and a more “plug and play” manufacturing and synthesis of Many customers remember our principle as we continue to see the stationary phases, for example, from integrators, which offered an easier industrialization of instrumentation. 10–20 μm and larger particles down and more reliable processing of Combined with improvements in the to sub-2-μm particles in LC and the obtained data. specificity of detector systems, this fused silica capillary columns for GC As a manufacturer of will lead to lower detection limits and contributed a lot to the acceptance chromatographs, our major improved sample throughput. and spread of chromatographic impact was to offer instrumentation We believe that mass techniques. that makes the most of the latest spectrometric and spectroscopic Last but not least, big developments in separation science. techniques will continue to flourish improvements on the detection side It was always our intention to and provide increasingly less extended the application range and make these instruments more reliable ambiguous data through improved offered new chances to use such and applicable for routine use. accuracy and sensitivity. If we systems: photo diode array detectors Supporting special developments, combine these advances with in LC, a variety of LC–MS and such as comprehensive LC×LC and improved data manipulation GC–MS detectors, and a GC×GC, in terms of hardware and techniques, we believe that the breakthrough in ionization techniques software is just one example where ability to elucidate even the most and practical implementation, we see our role having a big impact complex samples will become especially for LC–MS. For many on chromatography. increasingly simple. years, GC–MS was the “gold Recently, Shimadzu introduced These improvements in standard” in terms of high-sensitivity a supercritical fluid extraction/ instrumentation and software will systems, whereas nowadays supercritical fluid chromatography mean that the operator becomes LC–MS and GC–MS are seen more (SFE/SFC) system. We see this increasingly less likely to intervene as complementary techniques. as an interesting combination between loading up the autosampler Of course, each laboratory and of sample pretreatment and and reporting the result, which each analytical task has its own separation. Even without the use of of course takes us further away preferences. A special instrument to toxic organic solvents during the from the scientific processes that highlight in the field of detectors is extraction process, difficult and occur within the instrument. We will certainly the Orbitrap-MS. time-consuming extractions can be continue to develop services and Major impacts on the market performed faster and more efficiently. resources that enable analytical were the development of the scientists to understand the capillary GC columns and the How does your company see principles which underpin instrument release of ultrahigh-pressure liquid the current state-of-the-art of operation, software design, and chromatography (UHPLC) systems chromatography and, in this separation science in order to better (>10 0 M Pa). Both issues underlined context, what is the company understand what to do when things the necessity of renewing the doing to address unmet needs? go wrong, to spot when insidious complete system design by the State-of-the-art chromatography is problems are occurring, and to manufacturers and forced users very much driven by developments develop ever better methods to to reconsider existing “standard” in mass spectrometry. Standard serve their businesses. methods in terms of analytical chromatography matters, such as time, sensitivity, and suitability stationary phase development and — Anthony Taylor, Technical for state-of-the-art analytical technical aspects, seem to be less Director, instruments. important. Crawford Scientific Limited As well as the developments in Users in many application areas chromatography itself, thanks to the seem to be bound to existing development in PC technology and methods and techniques, which modern user interfaces for dedicated will probably remain the case for software, such systems have become quite a while, even though this more suitable for the operators and was predicted to change more surely supported the wide spread quickly. This also means that current Shimadzu of these techniques in modern developments are focused on just a laboratories. small group of users — which may Overall, what have been the be growing — while a larger group most important developments in What has been your company’s of analysts in routine laboratories chromatography over the past 60 highest impact contribution to gain less from the advantages of years (other than those made by chromatography over the past 60 those developments. For example, your company)? years? UHPLC was introduced years ago, Firstly, the realization of benchtop Instrumentation wise, Shimadzu but we still see a need to optimize instrumentation for routine laboratory started with GC 60 years ago and we the instrumentation in terms of use was the starting point for current have produced LC instruments for 40 robustness and system design. We chromatographs and forced a lot of years; we are proud of the release also see that current instrumentation related developments. of the photo diode array detector in has to be further improved to Secondly, improvements in the 1982 as one of the first suppliers. meet all requirements for ease of
20 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Past and Future Visions use, robustness, and flexibility. instrumentation compatible to the Ion chromatography (IC) has Improvements in current instrument ones we offer today. also improved. The single biggest control and data processing, and advance in IC was the idea of using if needed, on the concept of the — Björn-Thoralf Erxleben, Senior a suppressor to serve two functions: software, for example, would be Manager, Marketing Europe, the first to remove the eluent by necessary to enable truly “intuitive” Shimadzu a neutralization reaction to limit operation for software users. background and reduce noise, and From an end-user’s perspective, the second to convert analytes interchangeability of instruments into a more conductive form. This and data becomes more and more combination increases the sensitivity important. Instrument control by of IC, and today, all suppressors are another vendor’s software and data based on this fundamental concept. compatibility are also important Thermo Fisher Scientific There have also been advances issues. in GC; specifically the development Overall, what have been the of fused silica capillaries in the late Has your company thought about most important developments in 1970s dramatically improved the where chromatography might be chromatography over the past 60 performance and efficiency of GC. in 10 years’ time? If so, how might years (other than those made by The shift from single workstations the future look like? your company)? to enterprise deployments has Following the trends towards ease of The developments in simplified the chromatography use, faster separation, and high(er) chromatography over the past workflow and management of data. sensitivity detection, we see several 60 years have been immense. We aspects and challenges. have gone from primitive paper What has been your company’s We will see advances in chromatography, which was highest impact contribution to miniaturization of the system’s manual, time-consuming, and only chromatography over the past 60 hardware in parallel with a visually qualitative technique years? developments in using reduced flow to fully quantitated, automated, Thermo Fisher has made a rates and faster ways to achieve high-resolution, high-throughput number of contributions over the separations, including shorter chromatography in several forms. past 60 years to the different columns, smaller particles, new The basic principles of paper chromatography techniques. stationary phases, and coated chromatography have gone on to In HPLC, there has been a chips. Multidimensional systems spawn a range of chromatography significant industry shift in the last will become easy to use systems for techniques, each of which has seen 15 years to using higher pressure routine analysis. Sample preparation huge developments over the years. instrumentation for analyses. Thermo will be integrated into the analytical The development of UHPLC Fisher has played a large part in this process. is an example of where gradual transition and currently produces an Right now standard LC detectors improvements in chromatography instrument with the highest pressure are still UV-based (classic UV and both created and then overcame rating on the market, enabling rapid PDA) and in some areas still based limitations. Column chemistries analysis and improved resolution for on refractive index detection. In the have also advanced. The high complex samples. In addition, our future, MS techniques will definitely resolution form of HPLC came first zero-volume, fingertight fittings have play a more important role and new with an improvement in the column radically advanced ease-of-use in MS detectors may take over business chemistry, only to be limited by the standard and nanoflow HPLC. from the “standard” detectors, UV dispersion inherent in traditional In IC, the biggest contribution and PDA. HPLC systems. The low-dispersion by Thermo Fisher has been the There will also be an increase in UHPLC systems available now are development of the eluent generator. ready-to-use systems for specific essential to see the benefits of This development has truly analytical tasks. The appearance, high-resolution chemistries. revolutionized IC, as the automatic size of such systems, software In line with very narrow peak widths production of eluents provides control, and operation will be from gas chromatography (GC) and incredible ease of use, better different from current ones. We can UHPLC, the detector response rates detection limits, and the cleanest increasingly expect MS as standard had to be fast enough to collect possible eluent. detection in such systems, either as enough data points over a peak with Software, and the development of the only one, or as complementary a much decreased transit time in the the Thermo Scientific Chromeleon technique to highly specific detector. Electronics are now fast Chromatography Data System (CDS), detectors. enough to produce data collection is a marked contribution in the At the same time, established rates of up to 200 Hz. The ability chromatography software and data methods and approaches from to analyze thousands of samples management field. The system has today will remain, which means that each day produced the need for the now developed into a multivendor compatibility in terms of instruments automated high-throughput data software control for laboratories, and methods will play an important handling workstations we now see in the first to allow an analytical site role and force vendors to consider laboratories. to control all the instruments. When www.chromsoc.com 21 Past and Future Visions
integrated with our enterprise-level applications is a key driving force connections, which are fingertight informatics portfolio, including behind these demands. with near zero-dead-volume, allowing Thermo Scientific SampleManager Instruments now need to inexperienced users to set up nano LIMS, SDMS, and LES, it be essentially push button: flow rates. Developments in software increases laboratory efficiency easy-to-use and as user-friendly will further improve the ease of use for and productivity and connects as a smartphone, with the ability to chromatography systems, particularly the laboratory to our customer’s produce data at a high throughput in industries where compliance with enterprise. without compromising sensitivity regulations is crucial. In GC, the incorporation of and robustness. The incorporation high-performance MS directly of easy-to-use tools such as tablets, Has your company thought about into the gas chromatograph has as seen on instruments such as the where chromatography might be been a huge advantage to using Thermo Scientific Dionex Integrion in 10 years’ time? If so, how might high resolution accurate mass HPIC system, enables users to the future look like? spectrometry (HRAMS) because it intuitively interact with the instrument Chromatography will continue can help determine samples that to quickly obtain the data they need. to develop in many ways over haven’t yet been characterized. The introduction of automation the next few years, becoming an Detection of compounds of interest and the smart monitoring of increasingly powerful analytical tool. is another aspect where Thermo chromatography systems help In line with changing manufacturing Fisher has excelled. Universal laboratories reduce maintenance and laboratory needs, reducing detection with universal response and instrument downtime by flagging environmental impact and lowering was realized with the introduction when consumables need to be costs, the future will bring change of the charged aerosol detector changed and when the instrument and adaptability to accommodate in 2004. However, Thermo Fisher is conducting experiments under new and more efficient ways will always be remembered for conditions that are not ideal to of working. A need for more the introduction of Orbitrap MS, produce the best results, resulting in information at higher throughput first commercially introduced in optimal performance. in Quality by Design (QbD) 2005, which maintains a position of To look at resolution, a pushes the QC and production prominence as a technique for MS state-of-the-art chromatography divisions into the use of simple detection. system must have low dispersion and LC coupled to mass spectrometry In the area of sample preparation, high pressure capabilities to achieve (MS) assays to maintain control of breakthrough techniques such and maintain the resolution provided ongoing manufacturing processes. as accelerated solvent extraction by the modern chromatography Manufacturers will need to make (ASE) and autotrace automated media. The chromatography system these techniques amenable to SPE have significantly enhanced and column are integral to each other less-skilled operators, which will sample preparation workflows for and must both be up to the task. To involve simplified workflows and solid samples and liquid samples, achieve the goal of high-resolution, intuitive analysis software. respectively. More recently, chromatographic media have Another significant move that the Thermo Scientific SMART become smaller in particle size and is underway is the adoption of a Digest kit has enabled significant columns have become longer, both more holistic approach with greater improvements over conventional of which increase the demands support and collaboration in solving in-solution protein digestion on the pressure capabilities of the analytical problems rather than technologies. The kit reduces system. The new Vanquish Horizon simply just supplying the equipment. preparation times and associated UHPLC system increases the industry This has already been seen in errors, enabling fast, sensitive, standard upper pressure limit of clinical applications but will soon and reproducible results for UHPLC to 1500 bar. move to other markets too, such high-throughput applications. Sensitivity can come through as the biopharmaceutical industry, various approaches. The detector which has traditionally adopted very How does your company see of choice often reflects this, which complex workflows. For example, the current state-of-the-art of is why we provide a wide range peptide mapping involves a complex chromatography and, in this of detection capabilities from sample preparation step, for which context, what is the company fluorescence, electrochemistry, there is now a simple digest kit doing to address unmet needs? CAD, UV with light pipe technology, available. There are also several The requests of the analytical all the way to Orbitrap mass opportunities to remove sample community for improvements detection. To achieve higher preparation altogether and increase in chromatography have been sensitivity, one option is to reduce the information accessible with fairly consistent over the last few the scale of the chromatography. high-resolution intact MS using years: higher resolution, improved Nanoscale systems have proven Orbitrap technology. The use of sensitivity, faster analysis, and all essential in the field of proteomics. these global methods in analysis will with ease of use. The evolution of The advances that allow these reduce the amount of time taken in chromatography from a research systems to be used with the ease method development and increased tool used in a central facility to a of use expected from standard-flow automation of traditionally manual widely used instrument across many systems include nanoViper and often error-prone techniques
22 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Past and Future Visions will also be a driver for increased particles and bonded phases; the columns and deliver them on productivity. Software will continue first ion chromatograph in 1975; time, at an affordable price, and to play a critical role by turning data the first computer-driven robotic make a profit. Perhaps, most into knowledge and will adopt new sample preparation devices in 1984; importantly, Waters figured out how technology and delivery models, like the invention of the electrospray to educate and support customers the cloud, to accelerate processing source for mass spectrometry in the new technique. This strategy and simplify deployment. in 1984; the development of the became evident with the first GPC With regards to specific personal computer and the first Symposium in 1965. The strategy chromatography techniques, some chromatography data software was simple: get those who have great developments will be seen in for the PC in 1986; first sub-2-μm been successful with GPC together IC. The pressure rating is likely to particles in the 1990s; the in the same room with those who continue to increase, bringing with creation of the internet and on-line weren’t but who were motivated to it many performance gains. While commerce in the 1990s; first learn, and watch the magic take currently in the 10–15-min range, commercially-available LC systems place. in the future we could see analysis in 2004 built around sub-2-μm The early success with the times drop to the 30–60 s range, particles; and the emergence of GPC-100 and GPC-200 led Waters providing an instant information reliable and stable supercritical fluid Associates to introduce the stream. This will be particularly chromatography (SFC) systems and ALC-100, considered to be the first advantageous in many areas of columns employing 2.5-μm particle commercial HPLC system, in 1967. manufacturing that require a near technology in 2012. From 1967, HPLC improved continuous data output. incrementally until 2004, which Though the industry is prone to What has been your company’s is the year Waters introduced its change over the next 10 years, highest impact contribution to Acquity UltraPerformance LC (UPLC) Thermo Fisher will continue to chromatography over the past 60 System at Pittcon 2004 in Chicago, support the needs of industry and years? USA. This system became the first research, keeping aligned with future Scientists may not have known it commercially available instrument developments and listening to the then, but they were witnessing a built around sub-2-μm particles, for driving forces behind them. revolution in separations science which it earned a Pittcon Editors in the 1960s. The history and the Gold Award. Completely engineered — Jakob Gudbrand, President, growth of chromatography parallels from the ground up around the chromatography and mass the growth of Waters Corporation. column, the Acquity System proved spectrometry, Thermo Fisher From 1963 until Waters acquired the potential of sub-2-μm particle Scientific Micromass in 1997, chromatography column chemistries to increase was the company’s central focus. separation speed, sensitivity, and In 1963, Waters Associates resolution. Within five years, all introduced the GPC-100, an of the major instrument providers instrument that used the novel introduced systems of their own. technique of gel permeation chromatography (GPC), also known How does your company see Waters Corporation as size-exclusion chromatography the current state-of-the-art of (SEC), to measure the molecular chromatography and, in this Overall, what have been the weight distribution of the most context, what is the company most important developments in common polymers in use at the time doing to address unmet needs? chromatography over the past 60 — thermoplastics. Many consider Let me give you three good years (other than those made by this instrument to be the first high examples of how Waters is your company)? performance liquid chromatography meeting the unmet needs of The history of chromatography system to reach the market. Others, the chromatography laboratory. cannot be written without a consider the first HPLC system to Among them is the need to nod to outside influences be the Waters Associates ALC-100 get chromatographic data on including the development of system introduced in 1967. exceedingly small sample volumes, the personal computer and the During this time, Waters the need to detect more potent internet. In a nutshell, the most Associates’ signature contribution therapeutics dosed a lower levels, important developments to to chromatography was to take and, when doing biomarker occur with chromatography over it out of academic and research research, the need to find and the past 60 years include the laboratories — where scientists identify metabolites, peptides, and commercialization of the first gas tinkered with homemade columns proteins at ultralow concentrations. chromatograph in 1954 (okay and instrumentation — and put This calls for innovation in the area 62 years); the first gel permeation it into widespread commercial of microscale chromatography. chromatography system in 1963; use. This took entrepreneurship However, microscale the first high performance liquid and investment capital, and the chromatography hasn’t been easy chromatography (HPLC) system in operational expertise to reliably to pull off: It requires incredible 1967; the launch of rigid, fully-porous manufacture instruments and patience, dexterity, and time. Very www.chromsoc.com 23 Past and Future Visions
few chromatographers have been and are designed with scan rates processing and interrogating data able to put it to good use. that are compatible with today’s in new ways. In the future we can The Waters ionKey-MS system was UPLC/UHPLC instruments. They expect to see chromatography designed to make scientists much use less energy: they can run become even more useful. more successful with microscale effectively at 464W, and generate From an economical and chromatography. Incorporating a substantially less heat and use environmental sustainability point ceramic substrate inscribed with a 85% less electricity than legacy of view, we can expect to see more 150 μm (i.d.) channel packed with products. Moreover, they operate down-scaling of chromatography. 1.7-μm particles, the product was on 100–240 V, so that, no matter For example, microscale developed to provide reproducible where they are used, little in the chromatography greatly decreases and robust ultrahigh-pressure LC way of modifications are required solvent consumption, which reduces (UPLC/UHPLC) separations, provide to fit it into an existing laboratory solvent storage and disposal up to 40× sensitivity improvement infrastructure. Since they don’t costs and lessens the impact over 2.1 m i.d. columns, and provide require a roughing pump for on the environment. Microscale plug-and-play ease-of-use, making normal operation, they can be chromatography, by virtue of its it accessible to laboratory personnel turned on or off at the push of reduced injection volumes, also has at all skill levels. a button unlike conventional the potential of increasing the ROI of The ceramic substrate is then mass spectrometers. These investments in mass spectrometry, encased in an injection-moulded detectors also operate without the given its ability to improve the housing containing a column heater, sample-specific or user adjustments sensitivity of mass spectrometry and electronic and fluidic connections, of typical mass spectrometers. Any lower the limits of detection. and an electrospray emitter. When chromatographer can consistently The possibilities are always there the iKey device is locked into generate the highest quality mass for more simplified operation and position in the source, all of the spectral data routinely, without increased automation of the entire electronic and fluidic connections the need for any special training sample preparation and sample to the mass spectrometer are made or expertise. Waters Acquity QDa analysis workflow and we think we’ll automatically, thus eliminating any Detector is an excellent example. continue to see improvements there potential variability. Finally, if one takes a hard look at along with further improvements The sample is then introduced the sample preparation protocols in in separation technology allowing to and separated in the iKey use today, they’ll see a number of higher speeds and/or resolution for and transported directly to the slow, multistep workflows in need of the analysis of complex samples, integrated emitter, which converts improvement. Until recently that was such as mixtures of intact proteins the eluent into an aerosol. The true of released N-glycan analysis. It (such as top-down proteomics). plume of fine droplets in the aerosol can take 24 h or more to profile the This could potentially be achieved is ionized, giving the droplets a glycans released from a monoclonal by further reducing particle size positive charge at which point antibody. With a new chemistry and and by developing new particle they enter the vacuum of the MS labelling technique, Waters RapiFluor- morphologies and surface where they are further separated. MS reagent and kit, labs can now chemistries. Because mass spectrometers are release and tag the glycans and Waters continues to support the flow-sensitive detectors where profile them in a little more than 1 h. need of industry for process analytics, signal response is proportional to a growing market that is still young. the amount of sample reaching Has your company thought about Driving innovation in this regard is the the detector per unit of time, the where chromatography might be need for just-in-time manufacturing lower solvent flow generates a finer, in 10 years’ time? If so, how might and for product release data less disperse electrospray plume the future look? obtained at-line or on-line, rather and smaller droplets. These small Separations science continues to than in the laboratory. Some day in droplets go through fewer Coulomb be at the core of everything we do. the near future, we can expect to Fission events and have a greater The demand for information that see chromatography in one form surface-to-volume ratio compared to only chromatography can provide or another make its way out of the larger droplets. A higher percentage has never been greater. Also, the laboratory and onto the production of analytes is ionized, resulting in a need for greater separations fidelity floor and play a central role in fine- greater sampling efficiency by the and sensitivity continues to push tuning and checking the quality and mass spectrometer. investment into instrument and purity of drugs, chemicals, foods, Another driving force is the desire chemistry R&D. and beverages by providing real-time for ease of use, more information Resolving and characterizing information for batch or for continuous per chromatographic separation, more analytes per unit of time — manufacturing (flow chemistry) accurate quantification of sample increased peak capacity — is the environments. analytes, and quality of results. This driving force behind two-dimensional is where modular mass detectors LC, microscale chromatography, — Michael Yelle, Vice President, designed for chromatography fit supercritical fluid chromatography Separations Technology, Waters in. These detectors are about the (SFC), solid core particle technology, Corporation size of an ordinary optical detector and new software algorithms for
24 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Meeting Review: Advances in Microcolumn and Related Separation Technologies
Paul Ferguson, President, The Chromatographic Society, UK.
The first meeting in The Chromatographic Society’s diamond anniversary year brought together world-renowned speakers alongside past presidents of the Society, delegates from academia and industry, and scientific instrument and technology companies at the Institute of Engineering and Technology in central London on 22 March 2016. The presentations included discussions on liquid chromatography (LC), supercritical fluid chromatography (SFC), and gas chromatography (GC) from both an academic and industrial perspective — providing excellent insight into the state-of-the-art in these techniques for the delegates.
The Chromatographic Society’s analytes by gas chromatography (GC) Lynch, Peter Myers, Guy Wilson, and Advances in Microcolumn and by Pat Sandra; the diverse application Kevin van Geem. Related Separation Technologies was of supercritical fluid chromatography As part of our celebrations, we held at the Institute of Engineering (SFC) and SFC–MS — particularly of would be interested in finding and Technology (IET) on a beautiful drugs of abuse in sports science — by out what readers believe are the Spring day in Savoy Place, in London, Jean-Luc Veuthey; and a review of most important breakthroughs in UK, overlooking the River Thames. comprehensive multidimensional LC separation science in the lifetime There were over 130 attendees by Peter Schoenmakers, including his of The Chromatographic Society. registered for the meeting including group’s work in 3D chromatography Our proposals are listed in Table 1 14 vendor companies (including the for separation of complex peptide and more information for our principal meeting sponsors Agilent samples. This work on 3D-LC is reasoning may be found on The Technologies, Shimadzu, and Waters) a major endeavour that the group Chromatographic Society website presenting the latest from their is undertaking and comes under (www.chromsoc.com). product portfolios — all making for a the acronym, STAMP (separation To this end we are conducting a vibrant atmosphere. techniques for a million peaks). short on-line poll to ascertain your Some highlights included Ian One interesting aspect of Peter views, which can be accessed Wilson’s presentation of high Schoenmakers presentation was his through this link: https://www. throughput ultrahigh-pressure liquid thoughts on the 10 most important surveymonkey.co.uk/r/T7LBXTQ. chromatography–mass spectrometry breakthroughs in separation science We will publish the result early next (UHPLC–MS) metabolic profiling; an in the last 60 years. year in LCGC ’s The Column and on insightful discussion on approaches Other speakers included David the ChromSoc website. Please take for the analysis of thermally unstable McCalley, Frederic Lynen, Tom two minutes to share your views. There is an “other” option where you can suggest an alternative if you Table 1: In no particular order, the 10 most important advances or themes in separation don’t agree with what’s on our Top science in the last 60 years based loosely on some of Peter Schoenmakers thoughts at 10 list. the meeting. See our survey at https://www.surveymonkey.co.uk/r/T7LBXTQ to vote.
1. Comprehensive multidimensional chromatography (LC and GC) Paul Ferguson is the President of The Chromatographic Society. 2. Metal free spherical silica 3. Dual piston LC reciprocating pumps and gradient elution 4. Interfaces for hyphenation of LC and GC with MS detection A full review of the 5. Enantioselective separations Advances in Microcolumn and Related Separation Technologies 6. Sub-2-μm particles and UHPLC pumping systems meeting will be published 7. Capillary GC columns in LCGC ’s fortnightly digital 8. Reproducibly packed 1-mm i.d. LC columns and microflow pumping systems magazine, The Column, and The Chromatographic Society website 9. Solid core particles (both early and more recent variants) (www.chromsoc.com) 10. DNA sequencing using electrophoresis Photo credit: vladimir zakharov/Getty credit:Photo vladimir Images
www.chromsoc.com 25 ChromSoc Activities: The Reid International Bioanalytical Forum
Tony Edge1, Ray Briggs2, and Derek Stevenson3, 1Agilent Technologies, Stockport, Cheshire, UK, 2Eurofins Pharma Bioanalysis Services UK Limited., Oxford, UK, 3University of Surrey, UK.
The Reid International Bioanalytical Forum is a conference geared towards bioanalytical scientists. It has a unique philosophy where challenging data is often presented to an audience who are encouraged to engage. This article describes how the meeting series was initiated and developed over the past 40 years and how this philosophy has been embedded into the meeting.
Eric arranged the first set of Eric Reid was born in Aberdeen meetings in a style that was and studied chemistry at the unique to his own approach university from 1939 to 1943 before to “organizational excellence”. working at the Lister Institute This was part of the attraction of the meetings, as the informality Eric arranged the first associated with the registration set of meetings in a process and indeed the lecture slots fed very easily into the delegates style that was unique and the types of interactions that to his own approach to were often present within the forum. “organizational excellence”. This was further enhanced by the various social activities that often in London, UK, until 1946. He involved a degree of alcohol, but completed his PhD at UCL in 1949 always had a friendly atmosphere and then moved to Cambridge (with that was welcoming of both frequent F.C. Young), where he embarked visitors and those on their first visit. on a second PhD — Cambridge The meeting encouraged not only apparently not recognizing the one from UCL. In an illustrious The late Eric Reid, founder of The meeting attracted the academic career that saw Eric move The Reid International Bioanalytical best bioanalytical minds from the Chester Beatty Research Forum Institute, his studies moved towards and had a huge influence biochemistry, particularly nucleic The very first Reid International on the development of acid metabolism, subcellular, and Bioanalytical Forum was organized bioanalysis as a discipline. membrane biochemistry in normal by the late Eric Reid at the University and cancerous tissue. In 1964, he of Surrey, UK, in 1975. It was a great experienced presenters to come and joined the Battersea College of success and provided a launchpad share groundbreaking research but Technology where he founded the for an incredibly successful series were also a safe haven for first-time Biochemistry Unit. The award of the of meetings that have been held at presenters to develop and become Royal Charter in 1966 to Battersea Surrey ever since. The success of part of the community, including — forming the new University the meetings is down to the style of some of the present organizing of Surrey — was a moment of the lectures and the participation of committee. The tone and community celebration. In 1971 his department the delegates, whether it is in the of the meeting has fostered the moved to the current campus site intellectual confines of the lecture opportunity for people to present and a couple of years later, with hall or more likely the numerous some, not always, perfect data or the aid of a pump-priming grant social events that the committee scenarios — it has been easy to from the Wolfson Foundation, the has organized over the years. These present problems and say “we don’t Wolfson Bioanalytical Centre was events continued the intellectual know why” to get insights from the formed, with Eric steering it through debates and also fostered a community. The meeting attracted a very active, productive, and real community spirit within the the best bioanalytical minds and had academically successful period in meeting. This philosophy has a huge influence on the development the areas of analytical methodology been engendered by subsequent of bioanalysis as a discipline, often and subcellular biochemistry. It organizing committees to maintain leading the debate in important was during this period that the first the meeting in the ethos that Eric topics such as sample preparation Bioanalytical Forum was held in Reid had instilled from the start. and method validation. 1975.
26 The Chromatographic Society: Diamond Jubilee Supplement September 2016 The Reid International Bioanalytical Forum
carried on the Reid tradition stepped Figure 1: The Austin Pearce Building, the most recent University of Surrey home for the aside. The future for the Reid Forum is Reid event. still very bright. There are many more bioanalytical meetings globally than there ever has been and this gives competition for delegates, especially in the hectic and financially aware environment that we find ourselves in today. It should be stated though that these events can be seen as complementary, particularly when considering the approach that has been instilled as the ethos of the Reid Forum. In 2017 the Reid Forum will hold its 22nd meeting but for the first time at a new location, The Cambridge Belfry, on the 4–7 September but it will continue to encourage collaboration and debate in an informal setting. In addition to the main meeting, the organizers are offering training courses in bioanalysis and providing career The future for the Reid and his role in ensuring its success, advice to bioanalysts at its meetings. the organizing committee altered Forum is still very bright. the name to the Reid Forum, a very There were still the tales There are many more appropriate tribute to the great man. echoed throughout the bioanalytical meetings It was with great sadness that Eric passed away in 2010, a short time early morning sessions globally than there ever after the Forum had been renamed. of the night before antics, has been and this gives The following forum did miss with many legends being competition for delegates, Eric’s influence, however, the team of Ian Wilson, Howard Hill, Derek created in the confines of especially in the hectic Stevenson, Steve Westwood, Robin the university. and financially aware Whelpton, and Richard Houghton did an admirable job of ensuring The Reid International Bioanalytical environment that we find the continuing success of the event Forum is an event that was hewn by ourselves in today. and maintaining the ethos created the iron will of a man from Aberdeen, by Eric. There were still the tales which has seen the gradual With his wife Joan’s support and echoed throughout the early morning transformation into what is today a the recruitment of trustees, Eric sessions of the night before antics, suitable monument to his very ethos. started the Guildford Academic with many legends being created Associates (GAA), a charity in the confines of the university. Tony Edge is R&D Manager, devoted initially to continuing the Indeed, part of the attraction at that Polymeric Materials, Agilent Bioanalytical Conferences and time was that the accommodation CrossLab Group, UK. publication of methodological was very basic, but it did allow Ray Briggs is Head of Scientific surveys. GAA also endowed the for the after-party parties that saw Development, Eurofins Pharma “Guildford Bench Methodology Fund” the great intellectual minds of the Bioanalysis Services UK Limited., in within the Biochemical Society. bioanalysis world meet with the new Oxford, UK. Eventually the Chromatographic generation, thirsty for knowledge. Derek Stevenson was the Society took over the Bioanalytical This is absolutely the legacy that co-organizer of the Reid International Conferences in 1995, although Eric Eric has instilled within the forum, Bioanalytical Forum from 1995 to continued to have an input into the ensuring that the right atmosphere 2015 and visiting senior lecturer at organization of the event well beyond is created to allow this to happen. the University of Surrey, UK. this date. His academic career at Readers who have attended other Guildford ended in 1982, but the meetings will be aware that this focus of the GAA meant that Eric is not always the case and some The next Reid International was still very active, as well as being events can suffer as a consequence Bioanalytical Forum incredibly generous — donating the of too much formality. will be held at majority of his life savings to a range In 2015 the organization of The Cambridge Belfry of worthwhile causes. In recognition the event moved to a “younger from 4–7 September 2017. of his efforts to support the Forum generation” and the team that had www.chromsoc.com 27 High Impact Chromatographic Developments of the Past 60 Years
W. John Lough from the University of Sunderland, UK, in conversation with (old) friends from academia and industry to celebrate 60 years of advances in chromatography.
With the main theme of this publication technologies, etc. Also, we should issue and there we will see new being to celebrate 60 years of not forget it takes time to appreciate a developments, but there is a long developments in chromatography contribution as a revolution because way to go. The bottom line is that rather than just 60 years of The only the field (the users) will decide there is no standstill — certain Chromatographic Society, we thought whether or not it is a revolution. One application areas will significantly it might be useful to bring together can see this in very many areas. advance HPLC over the coming a small group of individuals, all of These developments are often years. These are related to life whom have been around long enough driven by challenging applications science analytics as to the various to have seen a good proportion of whereby these have also changed “omics” fields, biotechnology these 60 years. For the Society, John over the years. It is quite clear that fields (proteins, antibodies, etc.), Lough (JL) (Reader in Pharmaceutical at the beginning most of the HPLC and diagnostic areas. Synthetic Analysis in the Sunderland Pharmacy developments were driven by the polymers will get tackled in order School at the University of Sunderland, analytics of small molecules and of the to characterize them better from a Sunderland, UK) is talking to (in order pharmaceutical industry: now we are chemical point of view. of appearance!) Wolfgang Lindner shifting more and more towards large At this point it is worth noting that (WL) (Emeritus Professor, University molecules and proteins (RNA, DNA). this year two advanced European of Vienna and a Chromatographic This presents challenges on several Research Council (ERC) grants (ca. Society Martin and Jubilee Medal levels, including MS detection, which 2.5 million euros each) have been winner), Ted Adlard (TA) (former GC needs to be mastered. Although it has appointed to “chromatography”. One pioneer at Shell, a founding member for the potential of three-dimensional of The Chromatographic Society, and We should not forget it chromatography (Peter Schoenmakers Chairman 1970–1973, 1979–1983), [2]) and one for new silica-based Alan Handley (AH) (Senior Scientist takes time to appreciate a material using etching technologies at ICI and LGC, past President of the contribution as a revolution (Gert Desmet [3]). This is a clear sign RSC Analytical Division [2011–2014], because only the field( the about the importance of HPLC now and The Chromatographic Society and in the future. [2009–2014]), and Chris Bevan (CB) users) will decide whether (Big Pharma chromatographer, or not it is a revolution. Without the mastering Chromatographic Society President of selectivity issues, 2001–2007, and currently the been around for some time, the “slip Chromatographic Society’s Event flow” phenomenon explored by Mary multidimensional LC will Organizer). Wirth could offer new possibilities (1). not become routine in the Then we have the multidimensional future. The engineering of John Lough: Let’s be perverse LC×LC (LC×LC×LC) concepts, which and start with the here and now. continuously get better in order to the 2D-systems is already Wolfgang, it is timely that you substantially increase the overall peak at such a high level to the are just back from HPLC 2016 in capacity to deconvolute otherwise San Francisco. Is there anything overlapping peaks. This, however, point that the move is now exciting and new in separation builds on selectivity criteria, which to add further dimensions. science that we should be looking are in my view still one of the most forward to in eager anticipation? important issues for the success of JL: Well, that’s the present and the Wolfgang Lindner: I think we have the HPLC technology as a whole future all neatly wrapped up (for to face it: high performance liquid subject. Without the mastering of now) on a positive note. Having chromatography (HPLC) in its current selectivity issues, multidimensional LC said that, I can certainly support status is based on evolution rather will not become routine in the future. your provisos on selectivity issues. than on revolution. There were always The engineering of the 2D-systems There is still a lot to do on this. incremental improvements based on is already at such a high level to the Anyhow, let’s go back 60 years or theoretical predictions but also on point that the move is now to add so and have a think about how we technological improvements in material further dimensions. got here. In the beginning there science, engineering, software, Last, but not least, the material was packed column GC.
physics, mass spectrometry (MS) science component remains an Ted Adlard: The mid-1950s saw Photo Credit: Volker Gugath / EyeEm/Getty Images
28 The Chromatographic Society: Diamond Jubilee Supplement September 2016 High Impact Developments the emergence of gas chromatography (GC) with packed columns but with capillary columns already on the agenda. There was a great deal of development work being carried out in industry (in particular petrochemical) working alongside academia. The 1960s saw major detector development with the introduction of flame ionization (FID) DR. A. MAISCH GmbH and electron capture (ECD) detectors, and glass capillaries started to take over from packed columns. The real game HAS ACQUIRED SEVERAL changer came with the development of the flexible fused PRODUCT LINES FROM silica columns in the late 1970s to early 1980s, which really brought robust GC technology to the masses. GRACE DISCOVERY SCIENCES: Alan Handley: My career started in the GC area in the early 1970s and I had first-hand experience of the impact of some of the GC developments mentioned by Ted, but MEDIA AND COLUMN BRANDS: was also party to the emergence of a number of other Exsil™ and Exmere™ separation technologies in my role supporting ICI’s growing ™ ™ polymer businesses. Adsorbosphere , Allsphere , ™ ™ ™ Gel permeation chromatography (GPC), or size-exclusion Brava , Econosphere , Platinum , chromatography (SEC) as it now seems to be called, VisionHT and required both instrumentation and column technology GreatSmart capable of being operated with “exotic solvents” and at ™ high temperature — to facilitate the dissolution of the newer (formerly GraceSmart ) polymer types. This saw the major developments of both the “universal” refractive index detector and porous silica MODcol® preparative and cross linked polystyrene column packings. chromatography brands, GPC has always been a very robust including Spring® columns, technique thanks in the main to the Multipacker® packing equipment, overengineering required to handle the Vydac preparative columns solvents used in such systems and it is still the mainstay of the polymer industry. packed with Vydac TP, MS, Denali and Everest. Like all separation technologies, once established, we see the driver for quicker or better separations and more All associated hardware, information-rich detection. From the 1980s onwards we replacement parts saw the refinement of the column technologies — smaller diameter GPC packings, mixed bed and multipore columns and packing services (reducing the need for multiple individual pore -size column sets), the development of multidetector systems involving refractive index/viscometry and light scattering, and the associated challenges of handling and interpreting such data. GPC has always been a very robust technique thanks in the main to the overengineering required to handle the solvents used in such systems and it is still the mainstay of the polymer industry. I did eventually succeed in building an instrument in the late 1980s to run in concentrated sulphuric acid for the characterization of some of the newer engineering polymers — this is what chromatography challenges are all about! Beim Brückle 14 As ICI’s interests moved to polymer formulation I 72119 Ammerbuch-Entringen, Germany suddenly found myself swept up in the early days of HPLC, the progress of which is adequately summarized by some P: +49 7073 50357 of the other contributors, but I still look back at my earlier F: +49 7073 4216 experiences with a coil pump, syringe injector, and a UV E: [email protected] detector — one of the first modular HPLC systems. W: www.reprosil.com I have seen techniques such as field-flow fractionation (FFF), supercritical fluid chromatography (SFC), capillary electrophoresis (CE), and hydrodynamic chromatography (HDC) come and go — some have seen a recent www.chromsoc.com 29 High Impact Developments
resurgence but others have stayed I used a PerkinElmer F11 and had no of the pharmaceutical analytical as niche techniques awaiting an particular liking for it. I often had niggling laboratory. (Yes — chiral LC really industry need, a technology challenge thoughts around the consequences was difficult at that time but at least (human genome), or for instrument of the hydrogen or air flame on the one of my chiral stationary phases manufacturers to reinvent them. FID not lighting or the hydrogen flame [CSPs] proved to be more successful sucking back. Neither of these needs as a chiral catalyst [4]). For me, Two very important and have concerned me, what was more during these years as a postdoc, obvious high impact important was that it gave me low, highly the most striking development was variable results when analyzing the the introduction of the dual piston developments in the black tars produced when following a reciprocating pump. It was my rite early days of HPLC were patented industrial process. It took the of passage as a chromatographer advent of HPLC and a summer spent to break my first sapphire piston the introduction of 5-μm at ICI, Blackley in Manchester to sort all rod on a spanking new instrument. I totally porous spherical of this out. Using a system very similar successfully installed the replacement silica particles and then to the one described by Alan, I was and never made the same mistake soon able to identify and quantify all the again on the Waters pump or on the of reversed-phase HPLC major components in every black tar I LDC Constametric III pumps I later using alkyl-bonded silicas. produced. My beloved 100 × 4.6 mm used in industry. i.d. Corasil (40 μm pellicular) column JL: Wolfgang, as I recollect, your with septum injector followed me around JL: Moving on from the first decade early career involved using GC in everywhere thereafter until 1986 when or so of HPLC, the high impact investigations related to tobacco. an over-zealous laboratory tidy-up was of the commercial availability Did your switch to being an LC held at Beecham Pharmaceuticals of successful broad spectrum researcher involve that technique while I was away attending HPLC in San chiral LC columns is undeniable. offering you far more options Francisco! In the pharmaceutical industry in what you were doing or was it transformed the way in which it simply that you shifted to a During these years as a chiral drugs were developed. For different application area? postdoc, the most striking me, the big breakthrough was WL: Yes, I started with GC during the introduction of Hermansson’s development was the α my PhD studies at the Institute of 1-acid glycoprotein CSP in Organic Chemistry at the University introduction of the dual 1985, commercialized by LKB as of Graz, Austria. However, shortly piston reciprocating pump. Enantiopac. At Beecham, we went after I switched to the Institute of from being able to separate one Pharmaceutical Chemistry (at the It was my rite of passage racemic synthetic intermediate same university) — also doing GC. as a chromatographer to into its enantiomers using the I very soon saw the limitations for break my first sapphire Pirkle Type-1A CSP to being able real-world GC applications so I to achieve chiral separations for became interested in HPLC, for piston rod on a spanking every racemic drug development theoretical but also practical reasons. new instrument. candidate. Would you agree, In 1973 I bought the first HPLC Wolfgang, that Enantiopac was THE machine from Hewlett-Packard (serial Two very important and obvious major breakthrough in chiral LC? number 3), which was actually an high impact developments in the early WL: As you know, in 1978 I went as instrument designed and built by days of HPLC were the introduction a postdoc to Barry Karger and there Hupe and Busch. From thereon in, of 5-μm totally porous spherical silica I introduced them to chiral ligand while I worked in GC and LC, LC particles and then of reversed-phase exchange chromatography (CLCE) via became my addiction. (Later I did HPLC using alkyl-bonded silicas, the design and synthesis of a chiral also capillary electrophoresis [CE] though somehow they both rather ion pairing (chelating) compound and capillary electrochromatography passed me by. When I moved to being added to the mobile phase (of [CEC].) My passion has always been Edinburgh to work for Dick Wall, a reversed phase system). It was a the application and the understanding an important but less well known lipophilic chiral tri-amine and not an of “selectivity in LC” and very early senior member of the Edinburgh amino acid derivative as developed on I saw the potential and need, chromatography team, I found that the by Vadim Davankov. (He patented particularly when I worked for Sandoz use of 5-μm totally porous spherical his approach in 1968 in Russia and (now Novartis) in Basel in 1973 and silicas was already commonplace developed the first fully synthetic 1974 on a sabbatical for six months. there. Then, by the time I had chiral stationary phase). The year JL: I too was old enough to have started completed my postdocs involving the 1978 was the start of my “chiral” out working with GC. Such was the use of chiral mobile phase additives career. passion of practitioners of GC for their with silica and using silica to prepare To be frank, it is very difficult instrumentation, and one in particular immobilized chiral stationary phases to trace to one individual the that was set up in the early 1990s by (CSP) to move to the pharmaceutical success story of enantioselective The Chromatographic Society: The industry, I found that reversed-phase chromatography (see Vadim [5]). Very Pye 104 Club. It wasn’t like that for me. HPLC was already the workhorse early on in the game there were also
30 The Chromatographic Society: Diamond Jubilee Supplement September 2016 High Impact Developments
S. Allenmark, G. Blaschke, S. Hara, first chiral column on the market resolve small molecules went through Y. Okamoto, W. Pirkle, and so on. and described in the literature. In a a hype but slowed down significantly. You are right, the AGP column of J. broader sense and from a chemical However, for large(er) molecules, Hermansson became very popular point of view, it very soon became CE techniques clearly have true quickly in the pharmaceutical area clear that the polysaccharide-based advantages over LC techniques, because it showed a remarkable CSPs would win the game. However, particularly since the hyphenation with broad spectrum of selectivity even today, there remain niches to MS/MS has been elegantly solved. for many drugs. Many important be tackled and there is still room for So, CE and CEC have found their decisions within the pharmaceutical further developments. currently unbeatable niche. The industry may have been based on robustness of these techniques in this analytical technology. From It very soon became clear the course of small molecule analysis a theoretical point of view one that the polysaccharide- is not super strong, so it was not could not extract too much of an based CSPs would win surprising that people shied away insight on molecular recognition. It from these methodologies in routine became the area of other scientists the game. However, even laboratories. who contributed very important today, there remain niches SFC became first choice papers to the advancement of (at least judged from a large “chiral” chromatography. Now the to be tackled and there pharma company’s perspective) protein-based CSPs are more or less is still room for further in preparative chromatography outdated — this is the way it goes. developments. for medium-polar and non-polar Preparative enantioselective compounds. Analytically, however, it chromatography became extremely JL: Of course it hasn’t all been is still not fully accepted, particularly important in modern drug about LC and GC. From time to in routine laboratories. The merits development and this was the time other techniques have risen to are not so convincing that people clear limitation of the protein-type prominence. Overall though, would would switch from (U)HPLC to SFC. CSPs. The bottom line was that the anyone subscribe to the notion For lipidomics applications, however, Enantiopac column may have been that CE, CEC, and SFC have been SFC seems to have become the for a short time THE first choice chiral under-achievers rather than high first choice of separation technique column to have in the pharmaceutical impact developments? (speed and efficiency possibilities are industry, but it was certainly not the WL: The CE and CEC methods to the drivers). Method Development Refined
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www.chromsoc.com 31 High Impact Developments
JL: Chris, elsewhere in this and were not fully separable by the UK for your series of prep publication you have championed HPLC... at least not in our hands LC meetings. Are there any truly the impact of the much more at the time (mid-1990s). Micellar groundbreaking developments in sophisticated data handling we CZE (MEKC) separated them very that area that we should be noting? now have at our disposal. However, nicely, but of course we could not CB: Our series of BIG PREP symposia as an awardee for novelty in scale up this miniaturized technique was founded initially on a desire to capillary techniques, would you so were unable to prep them out in mix the analyst’s know-how with those also like to say a word on behalf of meaningful quantities. This spurred of the synthetic chemist’s and allow CE, CEC, or SFC? us on to research and develop CZE– a forum for cross exchange of ideas, Chris Bevan: The separation MS hyphenation to allow structural technologies, and working practices. of natural and unnatural elucidation of these and other Many of my synthetic chemistry oligonucleotides was routinely only molecules that had been successfully colleagues at Hoechst achievable by micellar electrokinetic separated by electrophoresis. In Pharmaceutical laboratories thought chromatography (MEKC) at the time. some specific cases we were able that it was somehow not the right way It was considered one of the most to separate preparatively using ion to use HPLC to isolate compounds of significant challenges to analytical exchange HPLC on an ionPac column interest and felt that they had failed in chemistry ever perhaps: certainly a because the separation principles their ability to synthesize specifically to really tricky problem neatly solved parallel some of the mechanisms produce pure desired products in high by electrophoresis (6). Micellar of CE. yields. electrokinetic chromatography was We were working on isomeric cis/ used to resolve diastereomers of Technological advances trans penem antibiotics at the time oligonucleotides possessing several using novel nano-switching (early-1980s) and these compounds chiral unnatural phosphoramidate were very difficult to synthesize pure in bridges. These materials were not and flow management the desired isomeric form. I had been resolved by conventional liquid have been applied to solve measuring these isomers analytically chromatographic techniques. capillary switching in by HPLC and suggested that we could Technological advances using scale-up HPLC to isolate both isomers novel nano-switching and flow instrumentation developed in a pure form. We could then test them management have been applied to decode the human for activity or toxicity and progress or to solve capillary switching in reject them more quickly. Pressure instrumentation developed to decode genome — arguably the from the parent company in Germany the human genome (7) —arguably biggest challenge ever to (Hoechst AG) was mounting to get the biggest challenge ever to analytical chemistry! pure isomers for testing so I built a analytical chemistry! A multiplexed large-scale HPLC system from scratch freeze-thaw switching principle (8) I would argue that the and purified both cis and trans penem and a distribution network were most-significant-to-man application isomers using a kilogram of 10-μm used to manage flow and sample of CE is in the separation of proteins Spherisorb ODS in a 4-inch diameter transportation. and DNA and RNA to elucidate the self-built axial compression prep HPLC structure of the human genome. system. This saved a lot of time and For lipidomics applications, The impact of SFC and the desire to went on to become common practice. however, SFC seems to develop and apply it comes from the The notion of fail fast and fail cheap ease at which the volatile fluids can changed the way in which our chemists have become the first be depressurized and evaporated thought, and led to high throughput choice of separation and the high plate efficiency with screening and combinatorial chemistry technique (speed and which separated products can be making impure mixtures for activity isolated very rapidly. testing. efficiency possibilities are Supercritical fluid extraction has It is curious to look back and the drivers). been applied industrially for many realize that chemists were routinely years to decaffeinate tea and coffee, using gravity column low resolution These three techniques and their and it does this very well. Many would chromatography and thin-layer derivatives have made a particular argue that SFC should have predated chromatography (TLC) to help purify impact on chiral resolution and on HPLC, but this ignores the difficulties their products, but they thought that the separation of biomolecules and of controlling pressures in systems and HPLC was a sledgehammer to crack structurally complex molecules. the poor solubility of many compounds a nut and such sophisticated tools Notable examples from my own in supercritical carbon dioxide, thus were the province of the analytical experience at Glaxo were separating limiting preparative applications. It may laboratories. some rather unusual natural be argued that CO2 is a “green” solvent WL: Final point from me: I think the product-derived compounds isolated but one should examine the whole separation techniques have strongly from fermentation broths. These procedure for “ecological greenness”. matured over the years and so have squalestatin compounds had complex the chromatographers. We have learnt and extraordinary chirality (eight JL: Chris, to turn to you again, to use the most appropriate technique chiral centres) and isomeric structures you are also very well known in for a given analytical question. There
32 The Chromatographic Society: Diamond Jubilee Supplement September 2016 High Impact Developments is no game anymore over which AJH: I agree with many of the there is still new monies coming technique is the better one. I always comments and whilst we do into chromatography, albeit in advocated that a good analytical now have more robust technologies Europe, but it does demonstrate chemist has to know to play the piano available, there are still challenges that chromatography is still, after 60 (keyboard) of methodologies and has out there, particularly in the area years, worthy of future investment to know the best key to meet the right of large biological molecules and interest. tone. This is what we need to teach. It and complex matrices. This will JL: Thanks, and thanks everyone for is based on a chemical understanding require “breakthrough technologies” your thoughts. I’d say let’s reconvene of the molecules to be analyzed and and will need the further in 10 years’ time to reconsider our the matrix it is dissolved in together development of both separation thoughts, but that would definitely be with a fundamental understanding of and detection technologies if we tempting providence. separation sciences, including MS. are going to break “the million compound separation” and provide References I would argue that the measurement solutions “at (1) B.A. Rogers, Z. Wu, B. Wei, X. Zhang, X. Cao, O. Alabi, and M.J. Wirth, Anal. source”. most-significant-to-man Chem. 87(5), 2520–2526 (2015). Capillary technologies have (2) http://www.ti-coast.com/news-events/ application of CE is in the brought much to modern society, latest-news/385-coast-congratulates- increasing scientific understanding prof-peter-schoenmakers-with-his-erc- separation of proteins and advanced-grant.html DNA and RNA to elucidate and fuelling product understanding (3) http://we.vub.ac.be/en/prof-gert-des- and development. met-wins-%E2%82%AC25m-grant-map- the structure of the human A number of instrument companies body%E2%80%99s-molecules (4) S.A. Matlin, W.J. Lough, L. Chan, genome. are worth a mention: Hewlett Packard D.M.H. Abram, and Z. Zhou, Chem. (now Agilent) successfully brought Soc. Chem. Commun. 1038–1040 JL: You are certainly right the chromatography community (1984). there. One of the key (sic) robust and fully validated GC and (5) V.A. Davankov, in Advances in Chromatography, J.C. Giddings, E. agenda items for now and for HPLC instrumentation when they Grushka, J. Cazes, and P.R. Brown, the future of chromatography were needed; Shandon (now Thermo) Eds. (Marcel Dekker, New York, USA, is to ensure that we have the and Phase Separations (now Waters) 1984) Chapter 17. (6) C.D. Bevan, I.M. Mutton, and A.J. Pipe, trained chromatographers to brought us high efficiency packings; Journal of Chromatography A 636(1), do and further develop the and Waters, who have brought 13–123 (1993). chromatography. However, to much to the table, gave us a (7) H. Tan and E.S. Yeung, Anal. Chem. 70(19), 4044–4053 (1998). round off the conversations “paradigm shift” with the launch (8) C.D. Bevan and I.M. Mutton, Journal of I would like to return to the of UPLC, which I feel reignited Chromatography A 697(1–2), 541–548 actual chromatography. What many people’s interest again in (1995). will we be working on and what chromatography. (9) See also: C.H. Arnaud, Chemical & Engineering News 94(24), 28–33 (2016). chromatographic techniques will we be using? Indeed, to use I always advocated that a John Lough is a Reader in Wolfgang’s metaphor, what tones good analytical chemist Pharmaceutical Analysis in the will we be trying to find and what Sunderland Pharmacy School at keys will we be using? I know that has to know to play the University of Sunderland, in we have skipped over the past the piano (keyboard) of Sunderland, UK, and a member of decade, and having celebrated methodologies and has to the Executive Committee of The 50 years it would seem logical to Chromatographic Society. emphasize the past decade when know the best key to meet Wolfgang Lindner is an Emeritus celebrating 60 years, but the whole the right tone. Professor at the University of Vienna, UHPLC vs. fused core–shell issue Austria, and a Chromatographic has been played out at ChromSoc The UK contributed much Society Martin and Jubilee Medal meetings and in ChromSoc-related to the early development of winner. media for a long time now. It seems chromatography: we had major Ted Adlard is a former GC pioneer to be politically correct these days companies who themselves pushed at Shell, a founding member of to say that they both have their the boundaries of research and had The Chromatographic Society, and merits, that they will continue to chromatographers who understood. Chairman 1970–1973 and 1979–1983. come together, and that eventually Sadly, many of these companies Alan Handley is a Senior Scientist it will be all about even smaller no longer exist, and with UK at ICI and LGC, past President of the but totally solid particles and academia not specifically focusing RSC Analytical Division (2011–2014), slip flow. For me though, Waters on this subject area, I do have and The Chromatographic Society deserve credit for their Acquity serious concerns as to where we (2009–2014). system, which, at the very least, will produce the next generation of Chris Bevan is a Big Pharma provided the catalyst stimulus for chromatographers needed to take up chromatographer, Chromatographic an exciting period of growth in future challenges. Society President 2001–2007, and LC instrumentation and particle However, as mentioned in currently the Chromatographic technology. Wolfgang’s opening paragraph, Society’s Event Organizer. www.chromsoc.com 33 ChromSoc Activities: Medals
Chris Bevan1 and W. John Lough2, 1Meetings Organizer, The Chromatographic Society, UK, 2University of Sunderland, UK.
A recollection of The Chromatographic Society’s medals and medallists over the years.
To the worldwide chromatographic fraternity at large, The Chromatographic Figure 1: Clockwise from top left: Roger Smith receives his Jubilee Medal from Society’s Martin Gold Medal and the President Derek Stevenson in 1998; Jack Henion, seen here with medal presenter, Silver Jubilee Medal are undoubtedly John Lough (right) at Edinburgh, is awarded the Martin Medal in 2003; a glittering array of medal winners at the Microcolumn meeting in March; and the Martin Medal. the most visible and recognized activities of the Society. In these days of awards proliferation, they are renowned as being amongst the highest distinctions awarded for excellence in the separation sciences. Perhaps to some this may be because they appear to have been around since the Society was formed. However, while they do have a long heritage, the facts do not quite bear this out. The Society was founded in 1956 but the Martin Medal was not awarded until 1978, to Ernst Bayer and C.E.H. (Ted) Knapman. The Jubilee Medal — as its name implies — followed on four years later just before the Society’s Silver Jubilee in 1982. The first Jubilee Medals were awarded to R. Tijssen, K. Grob Jr, and P.J. Simmonds. When first instituted the Jubilee Medal was originally intended outstanding contributions to the A complete list of medal winners may to be in recognition of the outstanding advancement of separation be viewed on the Society’s website achievements of young up-and-coming science. (www.chromsoc.com) Winners from chromatographers in the middle of their t The Jubilee Silver Medal is awarded 2006 are listed in Table 1. career. However, this has now been to younger separation scientists As we write, the process for selecting partially modified and the requirements who have made major use of medal winners for 2017 is underway. The for each award have been defined separation science in their own process has always been thorough but more clearly: field, or to scientists who have made nowadays it has become a protracted t The Martin Gold Medal is awarded contributions of merit to a particular in-depth affair involving the votes of the to scientists who have made area of separation science. members and examination of publications and CVs. Only in this way can we ensure that the medals are awarded to those Table 1: The Chromatographic Society Medal Winners since and including the Golden Jubilee Year, 2006. scientists and technologists considered to be of the very highest quality by Year Martin Medal Winners Jubilee Medal Winners Chromsoc’s committee and its members. 2006 Jim Waters Peter Myers, Stephen Wren 2007 Ronald E. Majors, Johan Roeraade Melvin Euerby Chris Bevan is a former Group Leader of the physicochemical characterization 2008No medal awarded David McCalley, Luigi Mondello team at GSK and former President of 2009 Wolfgang F. Lindner Gert Desmet The Chromatographic Society. He is 2010 Peter W. Carr Roman Szucs now retired and works as Chromsoc’s 2011 Peter J. Schoenmakers No medal awarded Events Coordinator. John Lough is a Reader in 2012 Edward S. Yeung Monica M. Dittman Pharmaceutical Analysis in the 2013 Gunther Bonn, Frantisek Svec Fabrice Gritti Sunderland Pharmacy School at 2014Nobuo Tanaka Michael Lammerhoffer the University of Sunderland, in Sunderland, UK, and a member 2015 Pavel Jandera Mary J. Wirth of the Executive Committee of The 2016 Ian Wilson, Peter Myers Derek Stevenson, Emily Hilder Chromatographic Society.
34 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Impact of Chromatography on Life and Society
Tony Edge, Vice-President, The Chromatographic Society, UK, and Agilent Technologies, Stockport, Cheshire, UK.
A theme for The Chromatographic Society Diamond Jubilee Year is to increase public awareness of chromatography. Often the general public, and even chromatographers themselves, fail to appreciate the importance of chromatography. Here Tony Edge, The Chromatographic Society Vice-President, discusses just how much chromatography underpins everyday life.
From the development of separation it is a robust technology that achieved. This presents an obvious science the world of analytical can separate relatively complex challenge in space where ordering chemistry has been radically samples. One of the challenges that a different column is simply not an transformed in the type of analyses faces many separation scientists option. that can be performed. It affects is accessing the level of technical In general, when considering the many aspects of our lives, from support that is required to ensure that selection of a detector, the more the food that we eat and the the chromatographic instrumentation universal its applicability the better, medication that we use, to the better is working in an appropriate because it will allow for a greater understanding of life outside of our fashion. The challenge of finding number of analytes to be detected. planet. Without chromatography an appropriate service engineer is The first mission to Mars used a the world would be a less wealthy, amplified substantially when dealing thermal conductivity detector (TCD), less clean, and a less healthy with extraterrestrial installations. with the robustness and general place. So how is applicability chromatography Figure 1: As far back as the Viking missions to Mars, gas chromatography (GC) was of this type actually being used to help scientists gain a better understanding of the solar system and the of detector incorporated possibility of life on other planets. being a major into these many consideration. varied aspects However, there of our lives? are limitations In this article, on the type of examples of detectors that separation science can be used; are discussed thus, flame alongside how the ionization use of this critical detectors (FIDs) science affects are not typically our standard of used in space living and also our because of the understanding of consideration of our place within transportation the Universe. of the air and hydrogen gas as Space well as the more Exploration obvious risk of The Universe is one of the more Another one of the challenges operating a flame at distance. esoteric areas where separation that faces extraterrestrial installations Sample pretreatment is also science has been employed. is ensuring that the analytical something that needs to be Chromatography has been used technology used is appropriate for considered. In most laboratories, extensively in space to help look the sample. Selection of the correct weighing out a sample and mixing for the building blocks of life. As far column, temperature gradient, and with a solvent, sieving, and back as the Viking missions to Mars detector is critical for ensuring that analyzing the resultant supernatant in 1976, gas chromatography (GC) the analysis is successful, and this is is considered routine: doing was being used to help scientists very dependent on knowledge this remotely presents severe gain a better understanding of the of what the sample is. Thus, challenges. The application of more solar system and the possibility of perversely, having a good indication complicated pretreatments presents life on other planets. This analytical of what the sample is will actually even more challenging issues.
Photo Credit: rajeshbac/Getty Images rajeshbac/Getty Credit: Photo procedure was chosen because ensure that the best separation is However, there have been a range of
www.chromsoc.com 35 Chromatography and Society
different sample matrices that have applications of chromatography, is used wherever it is necessary to been analyzed in space including with an expert panel from industry determine an organic chemical, such soil, ice, and dust. representing separation scientists, as a poison in the body, or residues Another consideration for space which included Dr Leon Barron, of gunpowder, or indeed to try and analysis is that the size and weight Professor Apryll Stalcup, and identify the type of rubber used in a are absolutely critical parameters. Professor Andrea Sella. Beyond the car tyre, as was noted in the film My Thus the electrical supply and radio and moving into television, Cousin Vinny. Samples taken from capabilities of the instrumentation separation science has also been the scene of a crime may require have to be very carefully considered. seen in The Simpsons on several some form of sample preparation to More complex instrumentation will occasions, including when Martin convert the raw sample into a form require larger batteries, which will Prince gave a presentation on that is suitable for chromatographic require a heavier fuel payload. For Archer Martin. High performance analysis, or sometimes it is possible Mars, solar panels are effective liquid chromatography (HPLC) was to analyze the sample directly (for at generating electricity, but for also seen in Professor Jonathan example if it is suspected that distances further away from the Frink’s laboratory in another episode. the contents of a glass of water sun, this approach is not viable: Beyond the cartoon characters, have been contaminated with a examples here include the Rosetta chromatography can also be deadly poison). In general the and Cassini-Huygens missions. seen in the popular crime scene chromatographic technique is As experience grew with investigation (CSI) programmes. coupled to MS to aid identification extraterrestrial missions so the The creators of these programmes and quantification. complexity of analyses developed. Within the film industry, The use of GC coupled to Beyond the radio and chromatography has also been mass spectrometry (MS) was incorporated successfully to ensure demonstrated with the second moving into television, a viable plot. This was particularly Viking mission to Mars, and designs separation science true in the much acclaimed film, for using liquid chromatography has also been seen in Medicine Man, starring Sean (LC)–MS are very advanced. The The Simpsons Connery. Connery, who plays the use of LC in space is an exciting on several lead character, is seen as a loner development, but also presents occasions, including employed by a pharmaceutical many challenges associated with the when Martin Prince gave company to discover new drugs storage of the liquid and what to do from the Amazonian rainforest with the waste. a presentation on Archer where he has been sent. Connery’s It is quite often that a news Martin. predecessor is a local medicine reporter will state that NASA has man who has the cure for cancer, identified chemical reagents that do use technical help from a range which he demonstrates when a boy hint at life, but what is it that they of analytical and forensic scientists is brought to him with malignant are actually looking for? In many to ensure that the commentary neoplasms. Connery, along cases it is simple organic species associated with the text on these with another colleague from the and an understanding of the gas programmes is factually correct, pharmaceutical company, then goes composition. All of this information although there have been examples in search of the elusive component helps scientists to determine if where this has not always been the that can cure one of man’s deadliest there is life or the potential for life case. For example, trying to analyze enemies. This is initially focused on distant planets. However, one of the residuals of liquid nitrogen on the flowers the medicine man is the challenges that scientists face is may prove challenging. Some of seen using to treat the boy. Using contamination and there have been the laboratories shown are actual a gas chromatograph, Connery is too many false hopes of detecting working laboratories, with one of able to confirm that the flowers are life, which subsequently have been them based on the R&D centre of not in fact the source of the cure, found to be a contamination issue. Thermo Fisher Scientific in San Jose, but that the source comes from an To date chromatographic systems USA. endangered species of ants, who, have been sent to our moon (Apollo), In the real world, in addition with a degree of irony, get wiped out Saturn (Cassini-Huygens), comets, to their laboratory role, forensic by a subsequent fire after the cure (Rosseta), and to Mars (Viking, scientists will also testify as expert for cancer is found using the Curiosity). witnesses in both criminal and civil GC–MS. There are a few cases, and can work for either inaccuracies in the film, relating to Popular Culture the prosecution or the defence. the use of suitable gas cylinders Chromatography has been seen While any field could technically and identification of an unknown in many forms of popular culture be forensic, certain sections have compound, but in general the film in particular on radio, television, developed over time to encompass portrays an accurate reflection of an and also in films. On the radio, the majority of forensically related analytical laboratory. the UK’s BBC Radio 4 recently cases. So what exactly does However, it is not just the arts devoted an hour to the discussion chromatography have to do with where chromatography has found a of what chromatography is and the forensics though? Chromatography new home. In the world of finance
36 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Chromatography and Society
chromatography has also appeared, the world of separation science is Figure 2: Ex-Chancellor of the with an old-style HPLC system on required. Exchequer and Tatton MP George the back of the Bank of Scotland £20 Osborne focuses on pork fillets at the These approaches work to identify note. Many suggestions have been opening of Waters’s Mass Spectrometry the original drug and to identify if put forward as to which system is headquarters in Wilmslow, Cheshire, a suspect has been using a drug. being used is but to date none have UK, on 11th September 2014. Unlike in the films and various been officially verified. TV situational dramas, it is not The pumping system is possible to determine by taste what undoubtedly a gradient system using a particular drug is. For seizures two Waters M6000A pumps, but of drugs in their pure form, the the identity of the detector is less analysis is relatively simple and certain The chromatographer in the requires dilution of the sample in an image is Janet Mullen, who took over appropriate solvent before analysis from Betty Trafford as Wolfson Unit can be performed. Over the years technician in around 1977. Other the approach to drug analysis has Wolfson members at that time, other changed, with recent innovations than Knox himself, were Dick Wall, such as GC–MS and LC–MS/MS who played a principal role in the becoming the mainstream approach development of a would-be Shandon for confirmation. LC system, and post-docs John Done and Jadwiga Jurand. Others In the world of finance who worked with Knox outside the chromatography has Wolfson Unit were post-doc Andrew also appeared, with an Pryde and PhD student Forbes Mclennan, who were succeeded old-style HPLC system by Mary Gilbert and Harry Ritchie, on the back of the Bank respectively. of Scotland £20 note.
Chromatography is used the use of monosodium glutamate, Urine or blood sampling is wherever it is necessary which is used to enhance the taste typically employed when testing a to determine an organic of food but has little nutritional value. potential drug user and will require some form of sample preparation. chemical, such as a Drug Testing This could be to remove the matrix poison in the body, or Many of the drugs that the in blood or plasma samples or residues of gunpowder, or pharmaceutical industry have indeed a simple dilution for urine indeed to try and identify developed have a variety of analysis. Sample dilution is used side effects, some of which at ever increasingly when LC–MS/ the type of rubber used in higher concentrations can have MS is used as the analytical a car tyre, as was noted in a stimulating effect on the user. technique because of the specificity the film My Cousin Vinny. However, these drugs can be very and sensitivity of the technique. addictive and long-term use will Even when mass spectrometry is Food Analysis invariably result in adverse effects involved, the separation is required The largest industry in the world is for the user, and ultimately this could to minimize ion suppression effects the food industry, which is worth result in death. Substance abuse because it is essential that the data about 10% of the global GDP, or is widespread, with an estimated can be relied upon. about 10 trillion dollars. Food is an 120 million users of hard drugs essential requirement for life and, as such as cocaine, heroin, and other Newborn Screening such, it is critical that appropriate synthetic drugs. In 2013, drug The first interaction that most measures are put in place to ensure use disorders resulted in 127,000 people have with chromatography that the quality of the food is at a deaths, up from 53,000 in 1990. is the heel prick test, which was high level. Unfortunately, because of The highest number of deaths are developed by Robert Guthrie for the value of this industry, it is often from opioid use disorders at 51,000. the analysis of phenylketonuria, a targeted by criminals and cheaper Cocaine use disorder resulted in condition where the body cannot substitutes can be introduced to 4300 deaths and amphetamine use take in phenylanaline, resulting in improve profit margins. However, in disorder resulted in 3800 deaths. neurological damage. The initial test some cases these substitutes can Alcohol use disorders resulted was developed in the early 1960s be harmful to humans, such as when in an additional 139,000 deaths. shortly after Guthrie’s 15-month-old melamine was substituted for milk Governments around the world niece had been diagnosed with powder in China. In some cases the have taken steps to address the phenylketonuria, and used a substitutes are deemed to be legal high death rates associated with bacterial inhibition assay, but it was and in general these will affect the the abuse of drugs, but in order to soon realized that the approach
Image Credit: Courtesy of the Author quality of the food. For example, determine what an individual is using could be applied to the diagnosis
www.chromsoc.com 37 Chromatography and Society
of many other compounds. The test involves taking a spot of blood from Figure 3: Separation science is used to improve the performance of athletes in a legal a heel prick on the newborn onto a manner and also to detect illegal performance-enhancing drugs. piece of what is essentially blotting paper. This original sample can then be analyzed in a laboratory. Across the world many of the 134 million babies born each year are tested using the heel prick test. This type of analysis has developed and now many of the test compounds are analyzed using LC–MS/MS. In the UK the tests that use separation science include: This was very much the beginning of sports science and the thirst to get a better understanding of how the human body works would eventually lead to the involvement of Greece. The noted Ancient It is sad to note that separation separation scientists. Greek physician Galen (131–201) science, as well as helping improve wrote 87 detailed essays about the performance of athletes in a tQIFOZMBOBMJOF QIFOZMLFUPOVSJB improving health (proper nutrition), legal manner, is also employed tPDUBOZMDBSOJUJOFEFDBOPZMDBSOJUJOF aerobic fitness, and strengthening to allow illegal improvements in (MCADD — Medium chain muscles. Assyrian Hunayn ibn performances to be detected. acyl-CoA dehydrogenase Ishaq translated Galen’s work, The use of a range of drugs has deficiency) along with that of Hippocrates, into been noted by many athletes and tBNJOPBDJEQSPGJMF Arabic, which led to the spread of trainers to improve performance. tQSPQJPOJDBDJEFNJB Greek physiology throughout the However, these drugs are methylmalonic acidemia, and Middle East and Europe. This was dangerous and the long-term effects isovaleric acidemia very much the beginning of sports are inevitably detrimental to the tEJTPSEFSTPGUIFEJTUBMVSFB science and the thirst to get a better athlete concerned. cycle, such as citrullinemia, understanding of how the human argininosuccinic aciduria, and body works would eventually lead Conclusion argininemia to the involvement of separation These are just a few of the areas tNBQMFTZSVQVSJOFEJTFBTF scientists. that separation science plays a (MSUD) — Leucine New ideas upon the working and fundamental role. There are many tIPNPDZTUJOVSJBNFUIJPOJOF functioning of the human body other areas where separation t*TPWBMFSZMBDJEBFNJB *7" $ emerged during the Renaissance scientists are employed, but in this (Isovalerylcarnitine) as anatomists and physicians brief snapshot it is not feasible to tHMVUBSJDBDJEBFNJBUZQF (" challenged the previously known highlight them all. In a world that — C5DC (glutarylcarnitine) theories. These new scholars went craves information, it is analytical t-$)"%% -POHDIBJO0) beyond the simplistic notions of scientists that feed that desire, acyl co-A dehydrogenase the early Greek physicians and and amongst analytical scientists, deficiency) — C16OH (3-OH shed light upon the complexities chromatographers are without doubt palmitoylcarnitine). of the circulatory and digestive one of the leading resources. As the systems. Today chromatography is world develops, it is envisaged that Other tests are being developed an essential analytical tool to fully separation scientists will continue to to allow greater use of LC–MS/MS describe these processes and play a critical role in ensuring that technology. In this area the use a better understanding allows the world is healthier, cleaner, and of chromatography is key to athletes to train harder and to get wealthier. ensuring our children are fit and more from their training sessions, healthy. with careful monitoring of metabolic Tony Edge is R&D Manager, markers. Polymeric Materials, Agilent Sports Science 2016 is not only the 60th CrossLab Group and Sports science can be traced anniversary of the ChromSoc, but Vice-President of The back to its origins in Ancient is also marks another Olympic year. Chromatographic Society. Photo Credit: Chris Ryan/Getty Images
38 The Chromatographic Society: Diamond Jubilee Supplement September 2016 ChromSoc Activities: Academic Support
Karen Rome, Academic Support and Multimedia Secretary, The Chromatographic Society, UK.
A look at some of the opportunities ChromSoc currently offers.
Since 2010 The Chromatographic were honoured to receive a working in small to medium Society has provided over significant donation from his UK-based pharmaceutical and £34,000 in bursaries, which trust fund. In keeping with John’s biopharmaceutical companies have enabled many students outlook on separation science and who find it difficult to get full and scientists to deliver research and development of young financial support to attend scientific projects, attend international chromatographers, the John Dolphin meetings. symposia, and make important Fellowship was set up to support Through the John Dolphin contributions by presenting their PhD students who wish to present Fellowship the society has work. Collaboration is key to the their work at international symposia. supported 19 students to attend 15 success of science, research, and international symposia — here are innovation and in our partnership Student and Industrial some testimonials demonstrating the with other societies, such as the Bursaries benefits. British Mass Spectrometry Society To support the development of (BMSS) and the Royal Society of up-and-coming separation scientists Nicola Gray, LC–MS Technical Chemistry (RSC), we have provided we provide a number of bursaries Specialist, Shimadzu UK: funding to academics to deliver each year that enable students and “Receiving a student bursary from multidisciplinary research projects. scientists to attend a conference the Chromatographic Society Involvement of scientists at which benefits their research during my PhD enabled me to the outset of their careers builds or studies. Student bursaries travel to the US to attend the HPLC an appreciation of how robust are available to UK full-time 2012 conference, providing the chromatographic methods can students or full-time students at a opportunity to gain confidence in improve business outcomes recognized UK university. Industrial presenting my own research as and ensures separation science bursaries are available to young a poster. The whole experience maintains its leading role in global and novice chromatographers allowed me to broaden my scientific research. As such, the academic support is very Figure 1: Several portraits of John Dolphin can be viewed in the Society’s Golden important to us. The following is Jubilee brochure (www.chromsoc.com) but here we see John with fellow luminaries an introduction to the bursaries (left to right: Harry Ritchie, John Dolphin, John Knox, and Mike Cooke) in a familiar and funding we provide. For more group photograph taken at a Hypersil Young Chromatographer of the Year symposium. information or to apply please visit As well as the ChromSoc 60th anniversary, 2016 is also the 40th anniversary for our website: www.chromsoc.com/ Hypersil, the Knox-invented spherical, totally-porous silica high performance liquid academic-support.aspx chromatography (HPLC) packing.
John Dolphin Fellowships This fund is dedicated to the memory of John Dolphin (Figure 1), an eminent chromatographer (perhaps best known for Shandon Southern’s collaborations with John Knox’s Wolfson Chromatography Unit at Edinburgh University), founder of the Hypersil “Young Chromatographer of the Year” Award, and a long-serving member of our Executive Committee, including serving as Chairman (President in “new money”) from 1985–1988. Following John’s untimely death in 2009, The Chromatographic Society www.chromsoc.com 39 Academic Support
chromatographic knowledge, develop essential networking skills, Figure 2: A prophetic photograph? King’s College early-PhD student James Heaton and build lasting connections with chats to David McCalley at HPLC 2010 in Boston, USA. A few years later he would others in the field.” be working for David as a post-doc at the University of the West of England.
James Heaton (Figure 2), Senior Analytical Scientist, Pfizer: ”ChromSoc bursaries played a massive role in allowing me to travel to several international conferences. The impact of attending these symposia have led to me finding opportunities both as a post-doctoral researcher and more recently employment at Pfizer. I’m very grateful indeed to ChromSoc for all of their support and hope that future bursary recipients make the most of receiving these career-enabling awards.”
Since 2010 The Chromatographic Society has provided over £34,000 in bursaries, which have enabled many students spectrometry content, relevance to t"OFWBMVBUJPOJOUPUIFDSJUJDBM and scientists to deliver current topics, novelty, deliverables, factors that control the accuracy and development of the student. of gradient and isocratic research projects, Summer studentship projects reversed-phase LC retention time attend international have spanned many scientific predictions from isocratic and symposia, and make disciplines and used a wide range gradient input data, respectively. important contributions by of techniques. Topics have included: t%FWFMPQNFOUPGBOFXFYUSBDUJPO Involvement of scientists presenting their work. method for licit and illicit drugs for GC–MS analysis at the outset of their Sophie Bailes, Analytical R&D t%FWFMPQNFOUBOEPQUJNJ[BUJPO careers builds an Scientist, AstraZeneca: of a GC–MS method for the appreciation of how “Attending HPLC 2013 gave me the identification and profiling of new robust chromatographic opportunity to speak to leading legal highs experts in the field and listening to t4FQBSBUJPOPGPMJHPOVDMFPUJEFT methods can improve their presentations was a great boost using nanoLC–MS business outcomes and to my understanding. The whole t%FWFMPQNFOUPG ensures separation science experience was extremely positive nanoflow-compatible and I would highly recommend this chromatographic separations for maintains its leading role in to any PhD student.” longer, larger peptides generated global scientific research. by chemical cleavage Joe Russell, QC Chemist, t($o.4QSPGJMJOHPGIPOFZCFF Luke Robinson (Figure 3), BSc University of Cambridge: colony volatiles as an alternative BioPharmaceutical Sciences, “The bursary enabled me to present indicator of honeybee diseases University of Sunderland; 2016 my research at HPLC 2015 Geneva, t5IFVTFPGJPOJDMJRVJEDPMVNOTJO summer studentship recipient: to discuss my results with leading the separation of pesticides using “We had good hands-on experience researchers, and join a conga with GC–FID, GC–ECD, and GC–MS of Agilent 1290 systems at Level 2. Fabrice Gritti.” t .FUBMMPQSPUFJOQSPGJMJOHGSPNESJFE However, it was nothing like driving blood spots by SEC–HPLC– a system on a day-to-day basis. I Summer Studentships ICP-MS, for clinical application in might have expected this but I could To provide opportunities to students human health and disease never have imagined the other many and increase research and practical t'PMMPXJOHDPOUJOVPVTGMPX benefits. For example, I have had to skills in separation science, biotransformations by HPLC or develop skills on presentation of we joined the BMSS summer GC data and have been able to studentship programme in 2010. t4UVEZPGTUBUJPOBSZQIBTF appreciate the finer points of Studentships receive funding based retentivity as a variable in integration and other aspects of data on their chromatography or mass gradient LC peak capacity analysis.”
40 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Academic Support
scientific disciplines and having Figure 3: Luke Robinson at his UHPLC system at Sunderland, studying stationary good practical skills ensures the right phase retentivity as a variable in gradient LC peak capacity. result. ChromSoc members receive a discount on a range of training courses.
Karen Rome is The Chromatographic Society’s Academic Support and Multimedia Secretary. She is also a member of The Chromatographic Society Executive Committee.
For more information on any of the academic support we provide or to apply please go to our website: www.chromsoc.com/ academic-support.aspx Applications for summer research grant. This will be open studentships reopen in the first to academics at recognized UK If you require support for your quarter of 2017. universities, who have a separation research and would like further science-related research project and guidance please don’t hesitate ChromSoc Research will be available throughout the year. to contact us: Studentships chromsoc@meeting To expand on the opportunities Training makers.co.uk provided by the summer studentships Separation science is a technique the society are developing a new that underpins research across many
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Peter Myers, Department of Chemistry, University of Liverpool, Liverpool, UK
On the occasion of the Golden Jubilee of The Chromatographic Society, Peter Myers wrote “The Future of Chromatography” in The Chromatographic Society’s souvenir brochure. Now Peter revisits his chromatographic crystal ball to review his predictions and deliberate on what the future might now hold for separation scientists.
company, developing and manufacturing chromatography (HPLC) columns. In miniature UV–vis absorbance detectors 2006, 4.6-mm i.d. columns were still the based on patented technology”. most popular; today this is changing to Maybe my prediction failed because I 2.1 mm and 1 mm. Why is this taking didn’t foresee the growth of the mass so long? Research leaders have been spectrometer as the detector of choice shown the advantages of smaller internal and the reduction in price of mass diameter for over 10 years. Again I put spectrometers for chromatography. the blame back on the instrument and However, I am now starting to see a new column manufacturers. When using a move in liquid chromatography (LC) into smaller internal diameter column, more some instrument miniaturization and the care has to be taken to connect the small move to make portable LC instruments and even smaller capillary columns into Peter Myers ponders the future for field use. the instrument to avoid dead volume. from the University of Liverpool. But my main point was that I hoped we The fittings we use today are just not fit would have intelligent columns by now. for modern use. They are essentially the For the Golden Jubilee of the My argument was based on the fact that same format as the fittings used over Chromatographic Society in 2006, I the column is the heart of any separation 10 years ago with a frit and a modified wrote a small article on the future of system. The columns should therefore Swagelok fitting. We need new fittings chromatography. I thought it might be carry a smart chip not only to identify the with snap on or simple pressure fittings. nice to review this after 10 years to see if column but also to carry the method, so But again, we need the instrument and I got anything correct. that we no longer need to purchase a C18 column manufacturers to make a step In the 2006 article I talked about the column but a column for PAH analysis or change in their column or instrument new monolithic columns that offered a a column for basic drugs. The column will design. I do hope this will occur in the lower back pressure for the same plate be placed inside the instrument and it will next 10 years. But as you know, I have count as a classical particle-packed be the column that will set up the system been wrong before! column. I also discussed the introduction parameters, perform QC procedures So how do I foresee the future now? of sub-2-μm particles that offer higher and calibrations, and finally decide when Unfortunately, I don’t think changes are plate counts but at the expense of far it requires replacing. We haven’t seen going to come fast. But it could if someone higher back pressures. I asked the anything like this yet and we certainly have embraced the new nanotechnologies question “Are either of these the future of not seen new micro- and nano-fabrication using sensors (nanowires), electroosmotic chromatography? Well let me make my techniques to make columns such flow on controlled surfaces, and first bold statement and say no.” I think I that a whole range of functions can be electrically controllable bonded phases. was correct —particularly with monoliths integrated into a single column format. We have to get the instrument and and to a great extent with sub-2-μm I also tried to look forward into the real column manufacturers to move out of the particles. Today I still believe sub-2-μm future and in this I had hoped we would separation science box in which they are particles may be OK for the super clean see the end of the chromatography happy and profitable. I hope they will look samples found in the pharmaceutical column and the high pressure pumps at what is happening in some universities industry, but in the real world we still for analytical separations. I hoped we (especially Liverpool!). have dirty samples and these are still would have moved into nonmechanical For yourselves, read all about the new best handled by columns packed with pumping systems using electroosmotic sensors, materials, and flow induction 5- or 3-μm particles. flow. I was wrong. We are still using methods in the new journals and you will After these two points I’m afraid I pumps and general columns with see how we as separation scientists are got things wrong — well maybe not different bondings are still the centre of being left behind modern technology. wrong, just that there haven’t been the our separations. All that has changed I hope in another 10 years I can write developments I expected. For example, I is that we have smaller particles and “Do you remember when we used to use had hoped to have seen new on-column some different porosities. The pump is pumps and packed columns for HPLC?”! injectors and on-column detectors. still king. Unfortunately, I cannot foresee We still do not have these, but maybe a change to this, because today’s Peter Myers is currently a Visiting we are seeing a move to a new form instrument manufacturers are really Professor of Chemistry in the of UV detector through the work of pump manufacturers. Department of Chemistry at the Paraytec. Set up by David Goodall on We have, however, seen a small University of Liverpool, in Liverpool, UK, his retirement from the University of change in the reduction of the internal and a Chromatographic Society Martin York, Paraytec is “a scientific instrument diameter (i.d.) of high performance liquid Medallist.
42 The Chromatographic Society: Diamond Jubilee Supplement September 2016 Are you seeking regulatory approval for your glycosylated biopharmaceutical? /ĨƐŽ͕LJŽƵŶĞĞĚƚŽƐĂƟƐĨLJƚŚĞĞǀĞƌͲƟŐŚƚĞŶŝŶŐƌĞƋƵŝƌĞŵĞŶƚƐĨŽƌĂĐĐƵƌĂƚĞ͕ ĐŽŵƉƌĞŚĞŶƐŝǀĞŐůLJĐŽƐLJůĂƟŽŶĂŶĂůLJƐŝƐ͘/ĨLJŽƵĨĂŝůƚŽĚŽƚŚĂƚƚŚĞŶLJŽƵƌŝƐŬ ƐƵīĞƌŝŶŐĞdžƉĞŶƐŝǀĞĚĞůĂLJƐŝŶŐĞƫŶŐLJŽƵƌĚƌƵŐƚŽŵĂƌŬĞƚ͘
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