JOURNAL OF CHROMATOGRAPHY LIBRARY volume 64

chromatographya century of discovery19002000the bridge to the sciences=technologyedited by

Charles W. GehrkeProfessor Emeritus of Biochemistry, University of Missouri,Columbia, Missouri, USA

Robert L. WixomProfessor Emeritus of Biochemistry, University of Missouri,Columbia, Missouri, USA

Ernst BayerProfessor of Organic Chemistry, University of Tubingen,Tubingen, Germany

2001

ELSEVIERAmsterdam London New York Oxford Paris Shannon Tokyo

JOURNAL OF CHROMATOGRAPHY LIBRARY volume 64

chromatographya century of discovery19002000the bridge to the sciences=technology

The Evolution of Chromatography. The Bridge to the Sciences=Technology. Some of the early scientists who invented, rediscovered, and=oradvanced chromatography include: M.S. Tswett, L.S. Palmer, R. Kuhn, A.W.K. Tiselius, A.J.P. Martin, R.L.M. Synge, F. Sanger, S. Moore and W.B.Stein, and the Awardees in Chapters 2, 4, 5 and S-9, S-10, and S-11.Who is the next farsighted scientist?

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First edition 2001

Library of Congress Cataloging in Publication DataChromatography: a century of discovery 1900-2000: the bridge to the sciences=technology = edited by Charles W.Gehrke, Robert L. Wixom and Ernst Bayer.1st ed.

p. ; cm (Journal of chromatography library ; v. 64)Includes bibliographic references and indexes.ISBN 0-444-50114-2 (hc)1. Chromatographic analysisHistory20th century. I. Gehrke, Charles W. II. Wixom, Robert L.

III. Bayer, Ernst. IV. Series.QD79.C4 C4837 2001543.08909dc21 2001053230

British Library Cataloguing in Publication DataChromatography: a century of discovery 1900-2000: the bridge to the sciences=technology. (Journal of Chromatog-raphy library; v. 64)1. Chromatographic analysis History 2. ChemistsI. Gehrke, Charles W. (Charles William), 1917- II. Wixom, Robert L. III. Bayer, Ernst. IV. Journal of Chromatography5430.089ISBN 0444501142

ISBN: 0 444 50114 2

1 The paper used in this publication meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper).Printed in The Netherlands.

VPrefaceChromatography A Century of Discovery 19002000 The Bridge to the

Sciences=Technology, is a documentary of seminal events, developments, discovery,and history of chromatography in the 20th century that presents the beginnings andstory of chromatography which revolutionized the sciences for students, researchers,and science for the 21st century. Its central theme is the impact of chromatogra-phy as a branch of science on different facets of science analytical chemistry,instrumentation, biomedical-, environmental-, pharmaceutical-, and space sciences, etc.

Who are the Chromatography Award winners? They were nominated by theircolleagues, evaluated by their peers and recognized for outstanding contributions. EachAwardee was recognized as a recipient of one or more international or national Awardsfor their investigations in the separation sciences. The 13 most important Awards andtheir respective sponsoring professional societies are listed in the outline of Chapter2. We invited over 100 Awardees in Chromatography to write about their uniqueactivities and careers in their own words and to share how their advances have impactedthe science disciplines.

The book represents the combined thinking and contributions of many chromatog-raphers and colleagues. E. Bayer (Professor of Chemistry, Director of the ResearchCenter for Nucleic Acid and Peptide Chemistry, of the University of Tubingen) is Editorin Europe. Chapter 2 is a compilation of international and national award winners inchronological order (most since 1975) from 13 professional societies and associationsfor 19002000. Among the very early group (1900s1920s) were M.S. Tswett, L.S.Palmer and C. Dhere, who contributed fundamentally to liquid adsorption chromatog-raphy. From E. Lederer on (1931), classical column chromatography was widely usedacross Europe. R. Kuhn and L. Zechmeister of Europe and H.H. Strain of the USA werewidely recognized as pioneers in this field.

The 1940s saw the seminal work of A.J.P. Martin and R.L.M. Synge on liquidliquidpartition chromatography, which then was utilized by them in the development of paperchromatography. At that time, A. Tiselius systematized the various chromatographicprocesses. Ion-exchange chromatography also started in the 1940s, with the ManhattanProject group utilizing the new synthetic polymeric resins for the separation of rareearths. Soon after, S. Moore and W.R. Stein (in the 1950s) extended IEC to theseparation of amino acids and E. Glueckauf, in England, provided the interpretationsof the basis of IEC separation. W.E. Cohn at Oak Ridge used IEC for the analysis ofnucleic acids, and S. Moore and W.R. Stein, together with D.H. Spackman developedthe first instrument the amino acid analyzer. E. Cremer in Austria, J. Janak inCzechoslovakia, C.S.G. Phillips in England and A.A. Zhukhovitskii in the SovietUnion carried out pioneering work on gas adsorption chromatography, but the real

VI Preface

breakthrough was represented by the adaptation of the principles of partition to gaschromatography by A.T. James and A.J.P. Martin in 1952.

At least a score of prominent scientists made fundamental contributions to the devel-opment of chromatography. The reader is referred to 75 Years of Chromatography AHistorical Dialogue by L.S. Ettre and A. Zlatkis, which tells the stories of 65 pioneers.It is true, and most important, that chromatography in the 20th century revolutionizedanalytical chemistry and research problems across the sciences. The discoveries ofchromatography fundamentally changed investigation of naturally occurring substances.Before chromatography the key word was isolation of one or only a few compo-nents of a substance; thus to obtain it large amounts of materials had to be prepared.Chromatography allowed separation of all the components present and brought in theworld of microchemistry in analytical chemistry. The changes in the direction of theSciences, introduced by GC, PC, TLC, IEC, HPLC, and AC=SEC in the 20th centuryare paradigm shifts leading to many new discoveries.

Unique aspects: This centennial documentarys purpose is fulfilled by the Pioneersand Builders of Chromatography in their Seminal Concepts in chromatography.Each Awardee was asked to write about their scientific discovery(s), and activities inchromatography. Their presentations (Chapter 5) are preceded by a short biography andin some cases picture(s) of the Awardees, their research years, and the place of theirlaboratory accomplishments to fully reflect the advances they have made in the sciences.Thus, this book is dedicated to the Chromatography Awardees and is written by them.This is their Book!

Several in-depth feature chapters cover the Beginnings of Chromatography. M.S.Tswett, the inventor of chromatography, and several other early pioneers (Chapter1) are highlighted; a discussion of the contributions of several Nobel laureates isincluded (Chapter 2). An extensive bibliography on the History of the Evolution ofChromatography (Chapter S-8); a presentation of Major International Symposiasupporting chromatography as a Bridge to selected sciences is included (Chapter3). Also, well-known chromatographers have written special chapters on Supports,and Stationary Phases and Separations (Chapter S-11), followed by a chapter onParadigm Shifts in Science (Chapter S-9) and one on the early Evolution of ScientificInstrumentation (Chapter S-10). Chapters 17 are in the bound volume, and ChaptersS-7 to S-15 are on the internet at http://chemweb.com/preprint/.

Chapter 6 is a unique presentation of Chromatography Around the World detailingthe chromatography research in Japan, Russia, China, Latin America, The Nether-lands and other countries. Prominent chromatographers wrote these chapters from theirrespective countries.

Chapter S-14 presents 24 Future Chromatographers of the 21st century whoreceived their PhDs recently with their abstracts and selected references. This isfollowed by a projection and perspectives of 41 of the Award Winners into the 21stcentury on the impact of Chromatography the advances made in the many sciencedisciplines and the development of emerging technologies in the new millennium.New discoveries in the biosciences and medicine, agriculture, the environment andseparations technology in the 21st century will rely immeasurably on the 20th centuryresearch tools in chromatography and those yet to be developed.

Preface VII

This book is recommended for Students in the Sciences and Research, Chro-matographers at all levels: professional scientists; research chromatographers inacademia, government, and industry; science libraries in academia, industry and pro-fessional societies, historians and philosophers of science; and educators and studentsat both high school and university levels. This book describes Chromatography asthe Bridge a key foundation built in the 20th century for major advances anddiscoveries yet to come across the sciences of the 21st century.

CHARLES W. GEHRKEROBERT L. WIXOMand ERNST BAYER

(Editors)

VIII

Dedication

To our scientific colleagues:

Who are the pioneers and builders of chromatography.

Who have contributed to this historical compendium.

Who have provided scientific thought, experiments, evaluation and communicationsto the world at large, and who interact with leaders in communities, states and inter-national realms. Our sincere thanks are extended to the many accomplished scientistsfrom around the world who have graciously and diligently presented their researchfindings in their contributed papers. Their efforts have allowed this treatise to presentan international, and a comprehensive perspective of the field of chromatography. Inthis fast-developing science and technology, their research findings have and will, playimportant roles in the advancement of science in many disciplines.

To our respective research institutions:

Whether academe, government, corporate, institutes, and foundations that havesupported this and other scientific research enterprises.

To our respective family members:

Who have given freely of their tangible support and valuable suggestions.

Our thanks are also extended to the editors of Elsevier Science.

IX

Acknowledgements

The Editors of this book had the benefit of many discussions and advice from otherchromatographers. However, one scientist stands out, Leslie S. Ettre, who not onlycontributed four chapters, but has served valiantly on many policy issues, a variety ofspecifics and relationships, and as a friendly critic. With his multiple experiences aseditor and his wealth of earlier chapters, reviews, historical explorations and severalbooks, the knowledgeable reader will not be surprised that we, the Editors, havedeeply appreciated the above role. Chapters 5 and 6, in recognition of achievements inchromatography are a logical extension of the 1979 book by Ettre and Zlatkis titled 75Years of Chromatography A Historical Dialogue.

We, the Editors, have had many helpful conversations with and advice from D.W.Armstrong, V.G. Berezkin, P.R. Brown, T.L. Chester, C.A. Cramers, P. Flodin, G.Guiochon, S. Hjerten, Cs. Horvath, W.G. Jennings, R.E. Kaiser, K. Jinno, B.L. Karger,J.J. Kirkland, F.M. Lancas, K. Macek, P. Sandra, L.R. Snyder, R. Tijssen, K.K. Unger,C. Welch and Y. Zhang.

We have appreciated the persistence and thoroughness of Nicole Hininger, a Univer-sity of Missouri student, who served as a Library Research Assistant. Many valuablesuggestions and some difficult online searches were provided by the excellent ReferenceLibrarians at the University of Missouri: Brenda Graves-Blevins, Janice Dysart, Re-becca S. Graves, E. Diane Johnson, Paula Roper and Caryn Scoville. The Editors havereceived helpful input from the librarians of the Chemical Heritage Foundation initiatedby the (American Chemical Society and other sponsors, Philadelphia, Pennsylvania) andChemical Abstracts (American Chemical Society, Columbus, OH). Preparation of copyfor this book is in large part due to the excellent secretarial skills plus accuracy andpatience of Crista B. Chappell, Nancy Harrison, Cinda Hudlow, Cynthia Mercado, KaraSeidel, Valerie Wedel, and Kelly Willcut at the University of Missouri, Columbia, MOand University of Tubingen, Germany.

The Editors have warmly appreciated the graphic artwork by Sammae Heard, MUGraphic Artist, and the pen and ink drawings by Corrine Barbour, MU Graduate ArtStudent.

Research, preparation, writing and editing this book was supported financially by theUniversity of MissouriColumbia, Missouri, USA: Chancellor, Richard L. Wallace, Vice Provost Jack O. Burns, and their Office of

Research. School of Medicine, and Dean Robert Churchill. College of Agriculture, Food and Natural Resources, and Dean Thomas Payne. Department of Biochemistry, and Chair William Folk.

X Acknowledgements

Experiment Station Chemical Laboratories (Agriculture), and Director Thomas P.Mawhinney.

University of Tubingen, Tubingen, Germany. Analytical Biochemistry Laboratory (ABC Labs), Columbia, MO, USA and CEO

Jake Halliday.

XI

Editors

Charles William Gehrke was born on July 18, 1917 in New York City. He studied atThe Ohio State University, receiving a B.A. in 1939, a B.Sc. in Education (1941) and anM.S. in Bacteriology in (1941). From 1941 to 1945, he was Professor and Chairman ofthe Department of Chemistry at Missouri Valley College, Marshall, Missouri teachingchemistry and physics to World War II Navy midshipmen (from destroyers, battleships,and aircraft carriers in the South Pacific) for officer training. These young men returnedto the war as deck and flight officers. In 1946, he returned as instructor in agriculturalbiochemistry to The Ohio State University in 1967 receiving his Ph.D. in 1947. In1949, he joined the College of Agriculture at the University of MissouriColumbia(UMC), retiring in Fall 1987 from positions as Professor of Biochemistry, Managerof the Experiment Station Chemical Laboratories, and Director of the UniversityInterdisciplinary Chromatography Mass-Spectrometry facility. His duties also includedthose of State Chemist for the Missouri Fertilizer and Limestone Control laws. He wasScientific Coordinator at the Cancer Research Center in Columbia until 1997.

Gehrke is the author of over 260 scientific publications in analytical and biochemistry.His research interests include the development of quantitative, high-resolution gas- andliquid chromatographic methods for amino acids, purines, pyrimidines, major and modi-fied nucleosides in RNA, DNA, and methylated CAP structures in mRNA; fatty acids;biological markers in the detection of cancer; characterization and interaction of pro-teins, chromatography of biologically important molecules, structural characterizationof carcinogenRNA=DNA adducts; and automation of analytical methods for nitrogen,phosphorus, and potassium in fertilizers. He developed automated spectrophotometricmethods for lysine, methionine, and cystine.

He has lectured on gasliquid chromatography of amino acids in Japan, China, and atmany universities and institutes in the United States and Europe. Gehrke analyzed lunarsamples returned by Apollo flights 11, 12 and 1417 for amino acids and extractableorganic compounds as a co-investigator with Cyril Ponnamperuma, University ofMaryland, and with a consortium of scientists at the National Aeronautics and SpaceAdministration (NASA), Ames Research Center, California, and the University ofMaryland, College Park, MD.

Awards and honorsIn 1971, he received the annual Association of Official Analytical Chemists (AOAC)

Harvey W. Wiley Award in Analytical Chemistry. He was recipient of the Senior FacultyMember Award, UMC College of Agriculture, in 1973. Invited by the Soviet Academyof Sciences, he gave a summary presentation on organic substances in lunar fines at

XII Editors

Charles W. Gehrke and Robert L. Wixom. Photograph taken on the University of Missouri Campus,Columbia, MO, USA, July 2000.

Ernst Bayer. Photograph taken at the Symposium on Environmental Technologies in the Research Center ofPETROBRAS, Brazilian Oil Company, September 2000.

Editors XIII

the August 1974 Oparin International Symposium on the Origin of Life. In 1975, hewas selected as a member of the American Chemical Society Charter Review Boardfor Chemical Abstracts. Sponsored by five Central American Governments, he taughtchromatographic analysis of amino acids at the Central American Research Institute forIndustry in Guatemala, 1975.

Gehrke was elected to Whos Who in Missouri Education, recipient of the UMCFaculty-Alumni Gold Medal Award in 1975 and the Kenneth A. Spencer Award fromthe Kansas City Section of the American Chemical Society for meritorious achievementin agricultural and food chemistry, 19791980. He received the Tswett ChromatographyMemorial Medal from the Scientific Council on Chromatography, Academy of Sciencesof the USSR, Moscow, 1978 and the Sigma Xi Senior Research Award by the UMCChapter, 1980. In 1986, Gehrke was given the American Chemical Society MidwestChemist Award. He was an invited speaker on Modified Nucleosides and Cancer inFreiburg, German Federal Republic, 1982, and gave presentations as an invited scientistthroughout Japan, Peoples Republic of China, Taiwan, Philippines, and Hong Kong(1982 and 1987). He was elected to the Board of Directors and Editorial Board of theAOAC, 19791980; President-Elect of the Association of Official Analytical ChemistsInternational Organization, 19821983; and was honored by election as the CentennialPresident in 19831984. He developed Libraries of Instruments, an interdisciplinaryresearch programs on strengthening research in American Universities.

Gehrke is founder, board member, and former Chairman of the Board of Directors(19681992) of the Analytical Biochemistry Laboratories, Inc., a private corporationof 250 scientists, engineers, biologists, and chemists specializing in chromatographicinstrumentation, and addressing world-wide problems on pharmaceutical and envi-ronmental issues to the corporate sector. He is a member of the board of SPIRALCorporation, Dijon, France.

Over 60 masters and doctoral students have received their advanced degrees inanalytical biochemistry under his direction. In addition to his extensive contributionsto amino acid analysis by gas chromatography, Gehrke and colleagues have pioneeredin the development of sensitive, high-resolution, quantitative high-performance liquidchromatographic methods for over 100 major and modified nucleosides in RNA, DNA,tRNAs and mRNA, and then applied these methods in collaborative research withscientists in molecular biology across the world. At the 1982 International Symposiumon Cancer Markers, Freiburg, German Federal Republic, E. Borek stated that ProfessorGehrkes chromatographic methods are being used successfully by more than half of thescientists in attendance at these meetings.

His involvement in chromatography began in the early 1960s with investigations onimproved GC methods for fatty acid analysis. Gehrke is widely known for developinga comprehensive quantitative gas chromatographic method for the analysis of aminoacids in biological samples and ultra-micro methods for life molecules in moonsamples. This method was used and advanced in the analysis of lunar samples whenhe was co-investigator with NASA. In the 1970s, his major interests shifted towardsthe development of quantitative HPLC methods for the analysis of various importantsubstances in biological samples, especially the modified nucleosides in tRNA asbiomarkers in cancer research.

XIV Editors

Major research contributions:

Developed eight methods adopted as Official Methods by AOAC International(formerly Association of Analytical Chemists). Sampling, Ca, Mg, K(2), P(2), N(2).

First to develop and automate AOAC official chemical methods for fertilizers (1950s1980s)

First to discover quantitative GLC of total protein amino acids (1960s1970s), 45publications.

First to develop quantitative HPLC of total nucleosides in tRNA, mRNA, DNAs andrRNAs (1970s1990s), 31 publications.

First to use HPLCMS nucleoside chromatography in molecular biology (19871994), 23 publications.

First to use GLC and HPLC methods for metabolites in body fluids as potentialbiological markers (19711994), 54 publications.

First to use GLC in analysis of Apollo 1117 moon samples at ultra high sensitivitylevels (19691974), 10 publications.

First to propose a Lunar=Mars-Based Analytical Laboratory (19891999).

Books: (AuthorEditor)

1979 Author, in: L.S. Ettre and A. Zlatkis (Eds.) 75 Years of Chromatography A Historical Dialogue. Elsevier Science Publishers, Amsterdam, TheNetherlands, pp. 7586.

1987 C.W. Gehrke, K.C. Kuo and R.L. Zumwalt (Eds.) Amino AcidAnalysis by Gas Chromatography, three volumes, CRC Press, BocaRaton, FL; 19 Chapters by 29 authors (5 chapters by C.W.G.) 543 pp.

1990 C.W. Gehrke and K. Kuo (authors=editors) Chromatography and Mod-ification of Nucleosides, a three volume treatise published by Elsevierin the Journal of Chromatography Library Series addressing AnalyticalMethods for Major and Modified Nucleosides, Biochemical Roles andFunction of Modification, Modified Nucleosides in Cancer and NormalMetabolism, and A Comprehensive Database of Structural Informationon tRNAs and Nucleosides by HPLC, GC, MS, NMR, UV, and FT-IRcombined techniques, 1206 pp.

1993 C. Ponnamperuma and C.W. Gehrke (Eds.), Proceedings of the Ninth Col-lege Park Colloquium A Lunar-Based Chemical Analysis Laboratory,A. Deepak Publishing, Hampton, VA, 282 pp.

1997 C.W. Gehrke, Mitchell K. Hobish, Robert W. Zumwalt, Michel Prost andJean Degres, A Lunar-Based Analytical Laboratory (C. Ponnamperumamemorial Volume, A. Deepak Publishing, Hampton, VA, 329 pp.

19541995 Nine additional chapters and reviews in other scientific journals andbooks.

In 1989 and 1993, C.W. Gehrke and C. Ponnamperuma of the University of Marylandwere named co-principal investigators on a proposal to address the scientific technical

Editors XV

concerns of placing on the moon a chemical laboratory which would be automated,miniaturized, computer robotic-operated and would support NASA programs in thestudy of five aspects of the exploration of space: (a) astronaut health; (b) closedenvironment life support; (c) lunar resources; (d) exobiology; and (e) planetology.

Gehrke received the American Chemical Society National Award in SeparationsScience and Technology in 1999 and the American Chemical Society National Award inChromatography in 2000 and the Ohio State University Alumni Achievement Award in2001.

Robert L. Wixom, Co-Editor of this book, was born on July 6, 1924 in Philadelphia,PA. In 1947, he graduated with a B.Sc. in Chemistry from Earlham College, Richmond,IN. He conducted his graduate studies and thesis at the University of Illinois under theguidance of William C. Rose, receiving his Ph.D. in Biochemistry in 1952.

Wixom held teaching=research faculty appointments in the Department of Biochem-istry, School of Medicine, University of Arkansas (19521964) and the Departmentof Biochemistry, School of Medicine=College of Agriculture, UMC (19641992). Hetook year-long sabbatical=research leaves at Oxford University (19611962), Universityof Wisconsin (19701971), Massachusetts Institute of Technology (19781979), andthe Fox Chase Institute for Cancer Research (19851986). His 40 years of research(45 peer-reviewed papers, two reviews) and graduate teaching focused on amino acidand protein metabolism. He taught intermediate and advanced biochemistry to medicalstudents, graduate students in diverse departments and undergraduate students with avariety of majors. Wixom guided the Advanced Biochemistry Laboratory course atUMC for 20 years, which covered several experiments in chromatography and 15 yearsteaching a course on Biochemical Information Retrieval. He has received three teachingawards. He served as a Departmental Representative to the Graduate Faculty Senate(19801993) and its Chair (19891992); this included a key role in three major newuniversity programs. He officially retired in 1992 as Emeritus of Biochemistry, butcontinues many similar activities.

Reflecting other earlier interests, Wixom was the co-initiator of the UMC Environ-mental Affairs Council, served as their first chair for three years (19901994), andcontinues as a member during retirement. He initiated and served as senior Editor ofthe 1997 book, Environmental Challenges for Higher Education: Integration ofSustainability into Academic Programs. The preceding experiences served as theeducational background for his present role as Co-Editor of the book Chromatography A Century of Discovery 19002000.

Ernst Bayer was born on March 24, 1927 in Ludwigshafen=Rhein, Germany, wherehe also visited primary and secondary schools from 1934 to 1947. From 1947 to 1952,he studied chemistry at the University of Heidelberg, and made his master thesis inPhysical Chemistry. From 1952 to 1954, he completed his Ph.D. thesis under the adviceof Nobel Laureate R. Kuhn, Max-Planck-Institute for Medical Research, HeidelbergUniversity on the structure of hemovanadin, a vanadium compound occurring in marinetunicates.

In March 1955, after a short postdoctoral fellowship in Kuhns laboratory, Bayer was

XVI Editors

appointed as Director of the Department of Biochemistry of the Government ResearchInstitute at Geilweilerhof. At this Institute, Bayer had also the task to study the quality ofwine and new cultivated sorts of wine. He developed in 1956, gas chromatographic (GCmethods for flavoring substances in wines and other beverages, and demonstrated theuse of GC in many other areas of natural compound chemistry derivatives. Also in thisperiod, preparative GC with large diameter columns up to 10 cm was demonstrated tobe a useful tool, and in 1959 he published the separation of the pheromones of silk mothusing the insects as a specific detector. At Geilweilerhof, Bayer started his investigationson metal proteins and biomimetic selective sequestering of metal ions, which led tothe development of polymers for the selective enrichment of gold, uranium and copperfrom seawater. From 1958 to 1962, Bayer was appointed as lecturer at the Institute forOrganic Chemistry, University of Karlsruhe. He continued his work on various aspectsof GC, metal proteins, selective enrichment of metal ions, recognized the importance ofmetal chelation for the color of flowers and fruits (blue color of cornflower versus redcolor of roses), and isolated flavor components of various beverages.

In 1957, his monograph, Gas Chromatography was published in German, andsoon translated into English and Russian. This book became for many years a guide forusers of GC.

In 1962, he was appointed Professor and Head of the Department of OrganicChemistry, University of Tubingen, a position he held until his retirement in 1996. Atpresent, he is Head of the Research Center for Nucleic Acid and Peptide Chemistryof the University of Tubingen. From 1967 to 1971, he held also the position of adistinguished R.A. Welch Professorship at the University of Houston.

Bayer has pioneered in HPLC, hyphenation of separation methods with MS and NMR.He published in 1972 the analysis of complex peptide mixtures, and detected the inherentfailure sequences in solid phase peptide synthesis (SPPS), which led to optimization andacceptance of SPPS, and made HPLC as the standard method for control. In 1976, hereported the first HPLC of dansyl-amino acids with fluorescence detection and reporteddetection limits down to the lower femtomoles. This performance has not been muchimproved since then. In 1974, Bayer reported the method known as template chromatog-raphy, using specific interactions of oligonucleotides and peptides. He made some ofthe first studies on the structure of the stationary reversed phases using solid state NMR(Cross Polarization and Magic Angle Spinning techniques). He used pulse field gradientsto derive values for the local axial and radial dispersion coefficients and determined theamount of mobile phase percolating through the bed and of stagnant fluid.

Recently he has concentrated on miniaturized separation methods like capillaryHPLC, and capillary electrochromatography and their on-line coupling to MS andNMR. He designed an instrument for the alternative use for all capillary separationmethods (HPLC, CE, CEC). In 1998, he published a new MS detection method,called Coordination Ion Spray (CISMS), which is based on the on-line formation ofcharged coordination compounds of the analytes. Non-polar compounds like sugars,lipids, terpenes, and vitamins can be detected as silver, palladium or boron complexes,which are not sensitively detected with electrospray MS. It is obvious that not all ofthe contributions of Bayer, which are reported in his 550 original publications, can besummarized here.

Editors XVII

Bayer is internationally recognized with many honors for his research in differentareas:

1978 A.J.P. Martin Award and the Tswett Medal of the USSR Academy of Sciences,for outstanding contributions to the development of chromatography.

1981 British Petroleum Energy Research Prize and the Max-Bergmann Medal forPeptide Chemistry for the development of Chirasil-Val and its application tostudy racemization of peptides and amino acids.

1985 Philip Morris Research Prize, Men and Environment for the developmentof a thermocatalytic, biomimetic process to convert biomass and sludge to apetroleum like oil.

1986 Tswett Chromatography Award for research in chromatography.1989 First Class Merit Cross of the Federal Republic of Germany; in 1990, the

International Rheinland-Prize for Environmental Protection for development ofanalytical methods relevant for the environment, and the Richard Kuhn Medalof the German Chemical Society for his research in structure elucidation ofmetal proteins and antibiotics.

1993 Fritz Pregl Medal of the Analytical Chemistry Society of Austria.1994 Fresenius Prize of the German Chemical Society for his contributions to

analytical chemistry and the Grand Merit Cross of the Federal Republic ofGermany.

1997 Maria Sklodowska Curie Medal of the Polish Chemical Society for his contri-butions to natural compound chemistry, and in 2000, the MTE. Golay Awardfor his contributions to capillary HPLC, capillary electrochromatography andhyphenated techniques.

2000 Halasz Medal of the Hungarian Society.2001 American Chemical Society National Award in Chromatography.

Bayer has served in many positions in the German Chemical Society, as a member ofthe editorial staff in many scientific journals, in government and science commissions,as a member of the IUPAC analytical chemistry nomenclature commission and memberof committees of scientific congresses.

XVIII

Chapter Outline

Chromatography a Century of Discovery19002000

The Bridge to the Sciences andTechnology

The Pioneers and Builders of Chromatography

1. The Beginnings of Chromatography The Pioneers (19001960) Robert L. Wixom 1

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2A. The initiation in Switzerland and Russia . . . . . . . . . . . . . . . . . . . . . . . . 4B. Adsorption chromatography (19001950s) . . . . . . . . . . . . . . . . . . . . . . . 7C. Partition chromatography (1940s1950s) . . . . . . . . . . . . . . . . . . . . . . . 11D. Paper- and thin-layer chromatography (two forms of planar chromatography) . . . . . . . 15E. Ion-exchange chromatography (IEC) (1930s1960s) . . . . . . . . . . . . . . . . . . 17F. Chromatography of petroleum . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21G. The literature of chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . 23H. Integration of seminal concepts with chromatography leaders . . . . . . . . . . . . . . 26I. From the inventors to the builders of chromatography . . . . . . . . . . . . . . . . . . 27J. What is required to be one of the award winners? . . . . . . . . . . . . . . . . . . . 29

Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

2. The Builders of Chromatography Major Chromatography Awardsand the Award Winners Leslie S. Ettre 39

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40A. Nobel Prize in Chemistry by the Nobel Foundation (19481999) . . . . . . . . . . . . . 41B. National Award in Chromatography of the American Chemical Society (19612001) . . . . 43C. National Award in Separations Science and Technology of the American Chemical Society

(19842001) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

Chapter Outline XIX

D. A.J.P. Martin Award of the Chromatographic Society (19782000) . . . . . . . . . . . . 43E. M.S. Tswett Chromatography Award of the International Symposia on Advances in Chro-

matography (19741988) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44F. M.J.E. Golay Award in Capillary Chromatography of the International Symposia on Capillary

Chromatography (19892000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44G. Stephen Dal Nogare Award in Chromatography of the Chromatographic Forum of the

Delaware Valley (19722000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47H. The Keene P. Dimick Award in Chromatography by the Society for Analytical Chemists of

Pittsburg (19882000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48I. Silver Jubilee Award of the Chromatographic Society (19822000) . . . . . . . . . . . . 49J. Award for Achievements in Separation Science of the Eastern Analytical Symposium (1986

2000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49K. COLACRO Medal (19862000) of the Congresso Latino Americano de Cromatografia . . . 50L. Leroy S. Palmer Award of the Minnesota Chromatography Forum (19802000) . . . . . . 51M. M.S. Tswett Chromatography Memorial Medal of the All-Union Scientific Council on

Chromatography, Academy of Sciences of the U.S.S.R. (19781979) . . . . . . . . . . . 52

3. Major International Symposia Supporting Chromatography Leslie S. Ettre 55

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55A. International symposia on (gas) chromatography by the (British) Chromatographic Society

(19562000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58B. Symposia on gas chromatography organized by the Instrument Society of America (1957

1963) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59C. International symposia by the French Society G.A.M.S. (19611969) . . . . . . . . . . . 60D. International symposia on advances in chromatography (19631988) . . . . . . . . . . . 60E. International symposia on high-performance liquid chromatography (HPLC) (19732000) . . 61F. International symposia on capillary (gas) chromatography (19752000) . . . . . . . . . . 62G. Danube symposia on chromatography (19761993) . . . . . . . . . . . . . . . . . . . 63H. COLACRO Latin American congresses on chromatography and related techniques (1986

2000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64I. Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy (PITTCON)

(19502000) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

4. Chromatography The Bridge to Environmental, Space and BiologicalSciences Charles W. Gehrke (continued in Chapter S-12) 69

A. Early years of automated chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . 72B. Chromatography in environmental analysis over the last 30 years . . . . . . . . . . . . . 74C. Amino acid analysis gasliquid and ion-exchange chromatography 30 years . . . . . 76D. Chromatography in space sciences GLC and IEC of Apollo moon samples . . . . . . . 83

5. Prominent Chromatographers and their Research Seminal Conceptsin Chromatography=Separation Sciences Charles W. Gehrke,Robert L. Wixom and Ernst Bayer 99

A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107B. Seminal concepts and new discoveries . . . . . . . . . . . . . . . . . . . . . . . . 108C. Relation of seminal concepts and awardees . . . . . . . . . . . . . . . . . . . . . . 108

XX Chapter Outline

D. Prominent chromatographers (76 awardees) . . . . . . . . . . . . . . . . . . . . . . 109E. Summary: if Mikhail Tswett were alive today . . . . . . . . . . . . . . . . . . . . . 598

6. Chromatography around the World Charles W. Gehrke,Robert L. Wixom and Ernst Bayer 601

A. Chromatography in Japan Kiyokatsu Jinno . . . . . . . . . . . . . . . . . . . . . 603B. Chromatography in Russia in the 20th century Viktor G. Berezkin . . . . . . . . . . . 621C. Chromatography in China Yukui Zhang and Guowang Xu . . . . . . . . . . . . . . 644D. Development of chromatography in Latin America Fernando Mauro Lancas . . . . . . 659E. Chromatography in The Netherlands (University of Amsterdam) Hans Poppe, Peter J.

Schoenmakers and Robert Tijssen (see their References in Chapter S-13) . . . . . . . . . 670

7. Overview: Chromatography A New Discipline of Science Charles W. Gehrke, Robert L. Wixom and Ernst Bayer 687

A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687B. Attributes of modern chromatography . . . . . . . . . . . . . . . . . . . . . . . . . 688C. Chromatography in the near future . . . . . . . . . . . . . . . . . . . . . . . . . . 689

Permissions for Copyright Items 691

Author and Scientist Index (see separate complementary author=subject indexfor the Supplement on the Internet) 695

Subject Index 705

A New Discipline of Science

Chromatography Chapters S-7 To S-15 are on the Internet at Chem Web Preprint Server(http:==www.chemweb.com=preprint=).

S-7. Overview: Chromatography A New Discipline of Science Charles W. Gehrke, Robert W. Wixom and Ernst Bayer

S-8. Bibliography of Publications The History of the Evolution of Chro-matography Leslie S. Ettre

IntroductionA. Books and BookletsB. Journal Papers and Book ChaptersC. General

Chapter Outline XXI

D. The Precursors of Chromatography (Pre-Tswett)E. M.S. Tswett. His Life, Activities, and CorrespondenceF. Nobel Prize LecturesG. Evolution of Liquid ChromatographyH. Evolution of Gas ChromatographyI. Petroleum ChromatographyJ. Key Contributors to the Evolution of ChromatographyK. Evolution of Chromatographic InstrumentationL. Meetings, Associations, and Personal Recollections

S-9. Milestones and Paradigm Shifts in Chromatography Robert L. Wixom

IntroductionA. Nobel Awardees Who Advanced ChromatographyB. Nobel Awardees Who Used ChromatographyC. Natureof Paradigm ShiftsD. Paradigm Shifts in Chromatography:

a. Carotenoidsb. Other Natural Productsc. Chromatography of Amino Acids, Peptides and Proteinsd. Affinity Chromatographye. Chiral Chromatographyf. Supercritical-Fluid Chromatographyg. Instruments for Chromatography

E. Further Developments in Chromatographya. Size-Exclusion Chromatographyb. High-Performance Liquid Chromatographyc. Detectors in Chromatographyd. Hyphenated /Coupled /Tandem Techniques in Chromatographye. Women Scientists in Chromatography

F. Summary (see Chapter 1 for Pioneers in Chromatography)

S-10. Evolution and Instrumentation in Chromatography Leslie S. Ettre

IntroductionA. Gas Chromatography Instrumentation, Detectors and ColumnsB. Liquid Chromatography Pumps, Detectors and Columns

S-11. Advances in Chromatographic Column Technology Ernst Bayer,Walter G. Jennings, Ron E. Majors, J. Jack Kirkland, Klaus K. Unger,Heinz Engelhardt, Gerhard Schomburg, William H. Pirkle, ChristopherJ. Welch, Daniel W. Armstrong, Jerker O. Porath, Jan B. Sjovalland Charles W. Gehrke

IntroductionA. Supports, Stationary and Bonded Phases

a. Column Development An Abbreviated History Walter G. Jenningsb. Future Advances in Column Technology Ronald E. Majors

B. Contributions by Other Chromatographers

XXII Chapter Outline

S-12. Chromatography The Bridge to the Environmental, Space and Biolog-ical Sciences Charles W. Gehrke (a continuation of Chapter 4)

S-13. Chromatography around the World References for Chapter 6 aregiven in Chapter S-13; the printed volume has Japan (KiyokatsuJinno), Russia (Victor G. Berezkin), China (Yukui Zhang and GuowangXu), Latin America (Fernando M. Lancas) and The Netherlands (HansPoppe, Peter J. Schoenmakers and Robert Tijssen)

S-14. Future Chromatographers of the 21st Century Contributions by 24Younger Scientists

S-15. Chromatography for the Next Millennium: Continuing Discovery andEmerging Technology Perspectives by 41 Chromatography Awardees

Appendices 17 Robert L. Wixom and Charles W. Gehrke

1. Glossary of Common Chromatography Terms2. Deceased Chromatographers Recognition and References3. Main Current Periodicals Covering Chromatography Serial Books, Review Journals,

Research Journals4. Selected Earlier Chromatography Books (Pre-1980)

A. General, Earlier Chromatography BooksB. Planar Chromatography (Paper-TLC) BooksC. Liquid Chromatography Books Early HPLCD. Ion-Exchange Chromatography BooksE. Gas Chromatography BooksF. Size-Exclusion Chromatography BooksG. Affinity Chromatography BooksH. Chromatography Handbooks

5. Chromatography Books by Series (Post-1990)A. Journal of Chromatography Library Series of BooksB. Chromatographic Science Series of BooksC. Separation Science Series of BooksD. Books on Chromatography by the American Chemical SocietyE. Books on Chromatography by the Royal Society of ChemistryF. Books by the Chromatographic Society (UK)G. Chromatography Books by Subject Areas (Post-1990)

a. General Chromatography Booksb. Planar Chromatography Booksc. Gas Chromatography Booksd. Ion-Exchange Chromatography Bookse. Size-Exclusion Chromatography Booksf. High-Performance Liquid Chromatography Booksg. Affinity Chromatography Booksh. Electrophoresis=Capillary Electrophoresis, Etc. Booksi. Supercritical-Fluid Chromatography=Extraction Booksj. Chiral Chromatography Books

Chapter Outline XXIII

6. Published Chromatography Symposia (19941999)A. Symposia in Journal of Chromatography AB. Symposia in Journal of Chromatography BC. Symposia in ChromatographiaD. Recent and Future Chromatography Symposia (2000 and 2001)

7. Books on Methods Related to ChromatographyA. Laboratory Techniques in Biochemistry and Molecular BiologyB. Methods of Biochemical AnalysisC. CRC Series in Analytical BiotechnologyD. Methods in Molecular BiologyE. Methods in BiotechnologyF. Methods in EnzymologyG. Methods in Molecular MedicineH. Other Book References on Methods

Author and Scientist Index for the Supplement see the Internet Chem WebPreprint Server (http:==www.chemweb.com=preprint=)

Subject Index for the Supplement (Chapters S-7 to S-15)

XXIV

List of Contributors

The Editors are pleased that 125 living Awardees and contributors have responded toour request for their concise research presentations. Their presentations and institutionaladdresses may be found in Chapters 5 and 6. The contributors in Chapter 5 are presentedin alphabetical order. Similarly, biographical information and a thesis abstract for 24,mostly 1999 Ph.D. investigators, may be found in Chapter S-14. Both are listed inAuthor Index (d=deceased). For complete addresses see the contribution in Chapters 5and 6 of each scientist=contributor.

Daniel W. ArmstrongIowa State University

Ernst BayerUniversitat Tubingen

Viktor G. Berezkin, Corresponding author,Institute of Petrochemical SynthesisProminent chromatographers from

Russia:

Vadim A. DavankovLaboratory at the Institute of

Element-Organic Compounds

Boris V. Ioffe dState University, Leningrad (St.

Petersburg), RussiaAndrei V. Kiselev dState University of MoscowKarl I. Sakodynskii dKarpov Institute of Physical Chemistry,

Moscow

M.S. Vigdergauz dInstitute of Organic and Physical

Chemistry

Aleksander A. Zhukhovitskii dAll-Union Research Institute for Geological

Prospecting of Petroleum (VNI GNI)

M.I. Yanovskii d

Morton BerozaUSA

Gunter BlobelRockefeller UniversityJerald S. BradshawBrigham Young University

Phyllis R. BrownUniversity of Rhode IslandTom L. ChesterMiami Valley Laboratories, Proctor and

Gamble Corporation

Carel A. CramersLab. of Instrumental AnalysisJohn V. DawkinsLoughborough University

Heinz EngelhardtUniversitat Des Saarlandes

Leslie S. EttreYale University

Michael B. EvansUnited Kingdom

Per G.M. FlodinArtimplant AB

List of Contributors XXV

James S. FritzIowa State University

Charles W. GehrkeUniversity of MissouriJ. Calvin Giddings dUniversity of UtahRobert GrobAnal. Chem. Consultant

Georges GuiochonUniversity of TennesseeAndras GuttmanNovartis Agricultural Discovery Institute

Steven B. HawthorneUniversity of North DakotaFrederich G. HelfferichThe Pennsylvania State University

Jorgen HermanssonChrom. Tech. AB., Stockholm

Herbert H. HillWashington State University

Stellan HjertenBiomedical Center

Csaba HorvathYale University

Daido IshiiKumamoto Institute of TechnologyReed M. IzattBrigham Young University

Jaroslav JanikAcademy of Sciences of the Czech RepublicEgil JellumInstitute of Clinical BiochemistryWalter G. JenningsJ&W Scientific CompanyKiyokatsu Jinno, Corresponding author,Toyohashi University of Technology

Shoji HaraTokyo College of Pharmacy

Hiroyuki Hatano dKanagawa Dental College

Nobuo Ikekawa dNiigata College of PharmacyDaido IshiiKumamoto Institute of TechologyHiroshi MiyazakiNiigata College of PharmacyTsuneo OkuyamaTokyo Dental College

Shigeru TerabeHimeji Institute of Technology

James W. JorgensonUniv. of North CarolinaOlga Kaiser and Rudolf E. KaiserInstitut fur ChromatogrophieBarry L. KargerNortheastern University

Jerry W. KingNational Center for Agricultural Utilization

Research

J. Jack KirklandAgilent Technologies

Ernst G. KlesperUniversity of TechnologyJohn H. KnoxUniversity of EdinburghFernando M. Lancas, Corresponding author,Universidade de Sao Paulo

Other chromatographers from SouthAmerica:

Clyde N. CarducciUniversity of Buenos AiresRemolo CiolaRefinery Research CenterArmando M. MorenoUniversidad Nacional Autonoma de Mexico

Joaquim LubkowitzSeparation Systems, Inc.

XXVI List of Contributors

Milton L. LeeBrigham Young University

Hendrik LingemanVrije UniversiteitCharles H. LochmullerDuke University

James E. LovelockCombe Mill, UK

Karel MacekCzech Academy of SciencesRonald E. MajorsAgilent Technologies

Karin MarkidesUppsala University

Archer J.P. MartinNational Inst. Med. Res. (Ex)Michel MartinEcole Superieure de Physique et de Chimie

Industrielles

Daniel E. MartireGeorgetown University

Robert B. MerrifieldThe Rockefeller UniversityHiroshi MiyazakiKawasaki, Japan

E. David MorganKeele University

Milos V. NovotnyIndiana University

Janusz PawliszynUniversity of WaterlooWilliam H. PirkleUniversity of IllinoisColin F. PooleWayne State University

Jerker O. PorathUppsala University

Michel ProstSpiral Corp., Dijon, France

Jacques RijksThe Netherlands

Pat J.F. SandraResearch Institute for ChromatographyFrederick SangerCambridge University

Raymond P.W. ScottConsultant, USA

Gerhard SchomburgMax-Planck Institut

Robert E. SieversUniversity of Colorado at BoulderColin F. SimpsonUnited Kingdom

Jan B. SjovallKarolinska Institutet

Hamish SmallDow Chemical Co. (Retired)Roger M. SmithLoughborough University

Lloyd R. SnyderLC Resources Inc.

Jun SuzukiSoda Aromatic Co., Ltd.

Shigeru TerabeHimeji Institute of TechnologyToyohide TakeuchiGifu UniversityRobert Tijssen, Corresponding author,Universiteit van Amsterdam

Other co-authors from the Netherlands:

Hans PoppeUniversity of AmsterdamPeter J. SchoenmakersUniversity of Amsterdam

Klaus K. UngerJohannes Gutenberg-Universitat

Irving W. WainerGeorgetown University Medical Center

List of Contributors XXVII

Harold F. WaltonRetired, USA

Phillip C. WankatPurdue University

Christopher J. WelchMerck & Co., Inc.

Ian D. WilsonDrugs Kinetics Group - Zeneca

Robert L. WixomUniversity of Missouri-ColumbiaEdward S. YeungIowa State University of Science and

Technology

Yukui Zhang and Guowang Xu and PeichangLu

Chinese Academy of Sciences

XXVIII

Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . VDedication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . VIIIAcknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . IXEditors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . XIChapter Outline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . XVIIIList of Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . XXIV

THE PIONEERS AND BUILDERS OF CHROMATOGRAPHY

1. The Beginnings of Chromatography The Pioneers (19001960) Robert L. Wixom . . 12. The Builders of Chromatography Major Chromatography Awards and the Award Winners

Leslie S. Ettre . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393. Major International Symposia Supporting Chromatography Leslie S. Ettre . . . . . . . 554. Chromatography The Bridge to Environmental, Space and Biological Sciences

Charles W. Gehrke . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 695. Prominent Chromatographers and their Research Seminal Concepts in Chromatography=

Separation Sciences Charles W. Gehrke, Robert L. Wixom and Ernst Bayer . . . . . . . 996. Chromatography around the World Charles W. Gehrke, Robert L. Wixom and Ernst Bayer 6017. Overview: Chromatography A New Discipline of Science

Charles W. Gehrke, Robert L. Wixom and Ernst Bayer . . . . . . . . . . . . . . . . . 687

Permissions for Copyright Items . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691Author and Scientist Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 695Subject Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 705

OVERVIEW: CHROMATOGRAPHY A NEW DISCIPLINE OF SCIENCE

See on the internet at Chem Web Preprint Server (http:==www.chemweb.com=preprint=)

S-7. Overview: Chromatography A New Discipline of Science Charles W. Gehrke, Robert W. Wixom and Ernst Bayer

S-8. Bibliography of Publications The History of the Evolution of Chromatography Leslie S. Ettre

S-9. Milestones and Paradigm Shifts in Chromatography Robert L. WixomS-10. Evolution and Instrumentation in Chromatography Leslie S. EttreS-11. Advances in Chromatographic Column Technology Ernst Bayer, Walter G. Jennings, Ron

E. Majors, J. Jack Kirkland, Klaus K. Unger, Heinz Engelhardt, Gerhard Schomburg, WilliamH. Pirkle, Christopher J. Welch, Daniel W. Armstrong, Jerker O. Porath, Jan B. Sjovall andCharles W. Gehrke

S-12. Chromatography The Bridge to the Environmental, Space and Biological Sciences Charles W. Gehrke

Contents XXIX

S-13. Chromatography Around the World References for Chapter 6: in Part A Japan (KiyokatsuJinno), Russia (Victor G. Berezkin), China (Yukui Zhang and Guowang Xu), Latin America(Fernando M. Lancas) and The Netherlands (Hans Poppe, Peter J. Schoenmakers and RobertTijssen)

S-14. Future Chromatographers of the 21st Century Contributions by 24 Younger ScientistsS-15. Chromatography for the Next Millennium: Continuing Discovery and Emerging Technology

Perspectives by 41 Chromatography Awardees

Appendices 17 Robert L. Wixom and Charles W. GehrkeAuthor and Scientist Index for the SupplementSubject Index for the Supplement

1CHAPTER 1

The Beginnings of Chromatography The Pioneers (19001960)

Robert L. WixomUniversity of Missouri, Columbia, MO 65212, USA

CONTENTS

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2Possible approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3Nature of this chapter . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

A. The initiation in Switzerland and Russia . . . . . . . . . . . . . . . . . . . . . . . . 4B. Adsorption chromatography (19001950s) . . . . . . . . . . . . . . . . . . . . . . 7C. Partition chromatography (1940s1950s) . . . . . . . . . . . . . . . . . . . . . . . 11

C.1. Liquidliquid partition chromatography (LLC) . . . . . . . . . . . . . . . . . 11C.2. Gasliquid partition chromatography (GLC) . . . . . . . . . . . . . . . . . . 12

D. Paper- and thin-layer chromatography (two forms of planar chromatography) . . . . . . . 15D.1. Paper chromatography (PC) . . . . . . . . . . . . . . . . . . . . . . . . . 15D.2. Thin-layer chromatography (TLC) . . . . . . . . . . . . . . . . . . . . . . 15

E. Ion-exchange chromatography (IEC) (1930s1960s) . . . . . . . . . . . . . . . . . . 17F. Chromatography of petroleum . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21G. The literature of chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . 23H. Integration of seminal concepts with chromatography leaders . . . . . . . . . . . . . . 26I. From the inventors to the builders of chromatography . . . . . . . . . . . . . . . . . . 27

I.1. Other early chromatography leaders . . . . . . . . . . . . . . . . . . . . . 27J. What is required to be one of the award winners? . . . . . . . . . . . . . . . . . . . 29

Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

References for introduction . . . . . . . . . . . . . . . . . . . . . . . . . 32References for Mikhail S. Tswett . . . . . . . . . . . . . . . . . . . . . . . 32References for predecessors of Mikhail S. Tswett . . . . . . . . . . . . . . . 33References on Leroy S. Palmer . . . . . . . . . . . . . . . . . . . . . . . . 33References on other early followers of M.S. Tswett . . . . . . . . . . . . . . . 33References on partition chromatography (LLC) . . . . . . . . . . . . . . . . 34References for gasliquid chromatography (GLC) . . . . . . . . . . . . . . . 34References on paper- and thin-layer chromatography (TLC) . . . . . . . . . . . 35References on ion-exchange chromatography (mainly early investigators) . . . . . 35References for petroleum chromatography . . . . . . . . . . . . . . . . . . . 36References on the literature of chromatography . . . . . . . . . . . . . . . . 37

2 Chapter 1

References on From the inventors to the builders of chromatography . . . . . . 37References for summary . . . . . . . . . . . . . . . . . . . . . . . . . . 38

Separation is as old as the earth : : : J. Calvin Giddings

Unified Separation Science, 1991

INTRODUCTION

This chapter presents chromatography as a branch of science that bridges a centuryof science and discovery. Chromatography represents the premier analytical method ofthe 20th century for the advancement of a variety of disciplines of science. The evolutionof chromatography is depicted in the frontispiece as a bridge and shows some of theinventors the builders who advanced chromatography and who constructed thefoundation of chromatography for later sciences and technology.

To continue with J. Calvin Giddings thoughts [1], the cloud of dust and gasesgathered and coalesced to form our planet Earth. Metal ions separated forming crystalsand ores iron in brassy-yellow pyrite (FeS2) or hematite ore (Fe2O3), silicon inclear, hexagonal cross-section of crystallized quartz (SiO2), multi-colored wavelite witha radial fibrous structure (Al3(OH)3(PO4)2 5 H2O, red cinnabar (HgS) and lead in theshiny, gray, cubes of galena (PbS). The suns energy, captured in photosynthesis led tothe separation of a carbon-prevalent biosphere and an oxygen-rich atmosphere (21%);nitrogen was concentrated in the atmosphere (70%), but is also abundant in the proteinsand nucleic acids of plant-, microbial-, and animal cells. Much more recently, Homosapiens has learned to distinguish protein- vs. carbohydrate-rich foods, isolated naturalproducts for medical use (e.g., quinine for malaria, etc.), and extracted natural dyes fromconcentrated plant sources (blue indigo from Indigofera genus; yellow quercitin fromthe black oak, Quercus velutina; red carmine from the female cochineal insect, etc.).

After the Big Bang and evolution, mankind has traveled in time through the latestages:

AgriculturalRevolution

! IndustrialRevolution(1700s1800s)

! Scientific=TechnologyRevolution(1800s1900s)

! InformationExplosion (late1900s2000s)

Present-day culture and science rests upon the scientific traditions and cultures ofEgypt, Mesopotamia, Greece, Rome, Islam, China and Western Europe; however, wehave spiraled far beyond these roots [2]. Within this context and in the past century,chromatography and separation science have incubated and grown with a flourish.But how do we scientists place a handle a meaningful organizational structure onchromatography a body of knowledge that is now too large for many scientists tofully grasp?

The Beginnings of Chromatography The Pioneers (19001960) 3

Possible approaches

As stated in the preface, the goal of this book is to recognize the pioneers andthe builders of chromatography, their discoveries and their personal recollections.Interwoven in their contributions and in other chapters will be seminal concepts, and thedevelopmental events during the past century. By undertaking the above goal, we arealso embarking on writing a history of this branch of science. Several approaches maybe found for organizing a presentation in the field of history of science: Portrayal of the sequence of scientific discoveries and their interrelationships.

(Example: M. Florkin and E.H. Stotz (Eds.), Comprehensive Biochemistry, ElsevierPublishing Co., Vols. 134, 19721986.)

Scientific biographies of the leading members of the branch of science underreview. (Examples: L.S. Ettre and A. Zlatkis (Eds.), 75 Years of Chromatography A Historical Dialogue, Elsevier, 1979; or M. Florkin and E.H. Stotz (Eds.),Comprehensive Biochemistry Sections on Personal Recollections, Elsevier, Vols.3540, 19831997.)

Flow of major concepts and hypotheses in a branch of science, the evidence forthe same, and sometimes or even frequently, their modification or even disavowal.(Example: T.S. Kuhn, The Structure of Scientific Revolutions, University of ChicagoPress, Chicago, IL, 1962, 1970 and 1996.)

Schools of scientific discipline, usually led by a distinguished scientist guidinghis=her colleagues, postdoctoral associates and graduate students.The above examples suggest the merits of each approach. Indeed some treatises

include more than one approach. The Frontispiece Bridge emphasizes the crucialrole of the earlier scientists=chromatographers to recognize the distinctive merits ofchromatography, to build the scientific structure of chromatography and to enhance theconnection the bridge to other scientific disciplines. Last, but not least, thesescientists inspired others to continue to construct bridges. Since chromatography is arelatively young branch of science, this book emphasizes the scientific biographies ofinternational and national awardees and contributors in chromatography and separationscience. Some authors of other chromatography books have a brief historical descriptionfor their specific chromatography area that is being described. V.G. Berzekin has writtento us that this book is also the Bridge from the XIX to the XXI Century. Thus, let usbegin.

Nature of this chapter

This chapter is a brief sketch a beginning for the rest of the book, as each of thefollowing subsections, A to F, has many references that are also presented by L.S. Ettrein Chapter S-8.

Furthermore, the history of these six subject areas has an extensive number of booksthat are cited in Appendices 4, 5 and 7, whose subdivisions are partially parallel tothe outline of this chapter. For Chapter 1, the reference numbers for journal articles orbooks are in regular type, and cross-references to other chapters (C) or the Appendices

4 Chapter 1

(A) are presented at the end of each subsection in italics along with the chapter numberand subsection, i.e., C-4B refers to Chapter 4-B, S-7D refers to a chapter in theSupplement (http:==www.chembweb.com=preprint) and A-5G to Appendix 5-G. Eachmain chromatographer is followed by a See Chapter 5B plus a letter code that definestheir area of chromatography as described in 5B.

A. THE INITIATION IN SWITZERLAND AND RUSSIA

M.S. Tswett, circa1910.

No discussion of the evolution of chromatography in the20th century can be complete without dealing with the life andactivities of its inventor.

Mikhail Semenovich Tswett was born on May 14, 1872, inthe small northern Italian town of Asti. His parents were on anextended holiday in Italy and were traveling by train toward theLago di Maggiore, the beautiful lake in northern Italy. The triphad to be interrupted because of the condition of the mother whothen died soon after the birth of her son. His father took the infantto Switzerland and Tswett grew up there, first in Lausanne andlater in Geneva where he studied botany at the University. By thebeginning of 1896, he finished his doctorate thesis dealing with

the structure of plant cells and chloroplasts and the movement of the protoplasma [1].Late spring of that year, he moved to Russia, joining his father who at that time was

Location of towns important in Tswetts life.

The Beginnings of Chromatography The Pioneers (19001960) 5

a high government official in the Crimea. Tswett had high hopes for an early academicappointment, but soon found that without a Russian advanced university degree thiswas not possible. Therefore, he moved to St. Petersburg and while having a temporaryposition in a laboratory, he carried out scientific research so that he could submit athesis and receive an academic degree. This was accomplished in the fall of 1901 atthe University of Kazan where he duly received a Russian Magisters degree. Soonafterward he moved to Warsaw, in Russian-occupied Poland, and in the next 14 years,he was active there, first at the University, then in 1907 at the Veterinary Institute, andfinally, from 1908 on at the Polytechnic Institute, although always in relatively minorpositions.

It should be noted that the controversy concerning the non-acceptance of his Swissacademic degree and then, for the next 15 years, the impossibility for him to obtain asenior university appointment made Tswett very bitter. He considered this discriminationbecause of his foreign background; as a conclusion, he became a loner and reacted veryharshly even to the mildest criticism. His strong polemic nature became a serioushandicap in his professional life and was justly criticized when, in 1918, he wasnominated for the Chemistry Nobel Prize [2].

Tswetts thesis work for the Magisters degree dealt with the physico-chemicalstructure of plant chlorophyll [3] and represented the start of his research whicheventually led to the development of chromatography. It is very intuitive to followthe successive steps of Tswetts investigations because these demonstrate his logicalthinking.

Tswetts aim was to isolate chlorophyll as close as possible to its natural state.During this work he found that while polar solvents (e.g., ethanol) can be used to extractchlorophyll from leaves, non-polar solvents (e.g., petroleum ether) are unable to do this.However, after chlorophyll was isolated from plants, it could be easily dissolved in thesesolvents. Tswett correctly concluded that this behavior is not due to simple solubilityproblems or to a chemical change of chlorophyll in ethanol to a soluble form, butis rather due to the interference of the molecular forces of the tissue, that is to say,to adsorption, and to the relative strength of the solvents compared to the adsorptionforces of the plant tissues.

The next step in his work was the study of the interaction of plant pigments with over100 different powdered organic and inorganic materials which may act as adsorbents,aiming to establish the general adsorption behavior of these substances. Eventually,these investigations led to a method permitting the separation of chlorophylls and somecarotenoids by stepwise selective adsorption and extraction. The results of these studieswere summarized by Tswett in a major lecture presented on March 21, 1903, in Warsaw,which was published two years later in a local scientific periodical [4].

From this method it was not far to realize that separation by adsorptiondesorptioncan also be carried out in a continuous way [5,6]. From 1903 to 1905, Tswettdeveloped this method and first referred to its existence (calling it only a new,reliable method, without giving any details) in two polemic publications criticizingthe results of Hans Molisch (18561937), then Professor at the University of Prague,on the pigments of brown algae. After being hard-pressed to present actual data anddisclose the way his investigations were carried out, Tswett finally submitted in June

6 Chapter 1

and July 1906 his fundamental two papers to the Berichte der deutschen botanischenGesellschaft, the journal of the German Botanical Society, published in the Fall.The first [7] dealt with his investigations of plant chlorophylls while the second [8]discussed in detail the new separation method developed by him, which he for the firsttime called chromatography. These two papers contain all the important aspects ofchromatography, including proper selection of the adsorbent, proper utilization of thesolvents and the possibility of using columns with different diameters (from 23 mm upto 1020 mm) and lengths.

These papers created considerable controversy and in the following years Tswettcarried out a lot of polemics with scientists in different countries who have worked inthe same field [9]. (For a detailed discussion of this controversy, see the papers by T.Robinson [10] and L.S. Ettre [11,12].) Meanwhile Tswett summarized the knowledgehe gathered on plant pigments, particularly chlorophylls, in a book published in 1910 inRussian [13]. A separate section of his book dealt with adsorption and chromatographyand in it, Tswett further expanded the description of the technique and its use, alsoemphasizing the possibility of preparative separations on columns having a diameter of30 mm and packing length of 80 mm.

Between 1906 and 1911, Tswett further expanded his research on plant pigments,including the carotenoids. It is practically unknown that this name for the polyenepigments used universally since the 1930s was first proposed by him, in a paperpublished in 1911 [14]. However, from 1912 on, he published almost nothing; his healthstarted to deteriorate and then came World War I with the interruption of normal life.Warsaw was occupied by German troops in the summer of 1915 and the PolytechnicInstitute was evacuated to Nizhnii Novgorod, but there was no possibility to carry outany research there. Finally, in 1917, Tswett was appointed a full Professor of Botanyand the Director of the Botanical Gardens at the University of Yureev (today: Tartu, inEstonia). He moved there in September 1917, but within a few months, German troopsalso occupied the Baltic area. A few months later, the Russian professors moved toVoronezh, in Russia, to start a new State University. Tswett was already very ill at thattime, and he died in Voronezh on June 26, 1919, only 47 years old. Thus, the life of oneof the most original thinkers of this period ended prematurely.

During his lifetime, Tswetts work on chromatography was not appreciated and wasbelittled by his peers. However, within a decade after his death, its importance wasfinally recognized and applied in almost every branch of science. Today, 80 years afterhis death, we consider chromatography as one of the most important inventions of the20th century.

By Leslie S. EttreSee Chapter 5B, a, b

Many other references on M.S. Tswett are available [15]. Though Tswett is generallyregarded as the father of chromatography, several other scientists have been identifiedas making brief entries in the 1800s into the general area of chromatography. Thenames and contributions of the predecessors of M.S. Tswett follow with appropriatereference(s):

The Beginnings of Chromatography The Pioneers (19001960) 7

Friedlieb F. Runge (17941867) was a German chemist who studied the spread ofcoal tar dyes on paper and made self-grown pictures for the Friends of Beauty;this area might be considered a precursor of the later (1940s) paper chromatography[1].

Friedrich Goppelsroeder (18371919) was a student of Friedrich Schoenbeim(17991867) and improved the latters approach for separation of dyes on hang-ing strips of unsized paper by capillary migration. Though he published a 1906monograph, and again in 1909, his work was ignored and then lost until the 1940s[2].

B. ADSORPTION CHROMATOGRAPHY (19001950s)

In adsorption chromatography, the molecules are separated on the basis of theiradsorptive properties, where the stationary phase is a solid adsorbent usually in acolumn or on a plate and eluted by the mobile phase that may be aqueous or organicsolvent(s). The adsorption chromatography research of M.S. Tswett was scoffed at bysome, but followed by others that are now presented. The individual whose research hadthe most impact on later investigators was Leroy S. Palmer.

Leroy Sheldon Palmer (18871944) was a pioneer in chro-matography research shortly after M.S. Tswett. Palmer was astudent at the University of Missouri-Columbia, Missouri, USA,earning a B.Sc. in chemical engineering in 1909 (a new depart-ment that began in 1903), and his M.A. in chemistry in 1910[1]. His Masters thesis was primarily an outline of his doctoralproblem and a detailed literature survey. Palmers investigationsleading to his Ph.D. degree began in October 1909 and werecompleted in the spring of 1913 in the College of Agriculture[2]. The full text of his thesis was published in four issues of theBulletins of the University of Missouri Agricultural Experiment

Station [1], and then in five successive research papers [3,4].Upon entering the Graduate School in 1909, he joined the Cooperative Government

Dairy Research Laboratory with Clarence H. Eckles (18751933), Professor and Chairof the Department of Dairy Husbandry (19011918). His thesis problem was toinvestigate the observations made by many dairy farmers, namely that butter from cowson summer pasture with fresh grass or green alfalfa hay has a deep yellow color,whereas cows consuming stored foodstuffs in winter produce a butter that has usually avery light color.

Just a few years earlier, M.S. Tswett had invented adsorption chromatographyand had separated the chlorophylls, carotenoids and xanthophylls from plant leaves(described more fully in the preceding paragraphs). R. Willstatter and W. Mieg [5]had just established the elementary composition of carotin and xanthophyll in1907, but their actual structures and various isomers were not deciphered until thelate 1920s. Thus Palmer had to make a choice in 1910 to isolate individual pigmentsutilizing their selective solubility and purify them in a multi-step process, ending in

8 Chapter 1

crystallization, or follow Tswetts method of adsorption chromatography; he chose thelatter. Palmer found Tswetts 1906 paper in the Berichte der deutschen botanischenGesellschaft, at a time when European scientists tended to ignore Tswetts observations.Relying on Tswetts style of adsorption chromatography, Palmer found carotenoidsin his extracts of butterfat and thus explained the change in color from summerto winter being due to dietary intake. Palmer, like Tswett, found that inulin andsucrose were suitable alternative adsorbents to calcium carbonate and used elution toremove the colored solutions, rather than extrusion as in C. Dheres laboratory. Healso used prefractionation of the pigments before chromatography, differential solventextraction and a crude spectrometric examination. His research represented probablythe first use of chromatography after Tswetts basic 1906 publications and introducedchromatography to scientists in the USA [3].

Palmer, after completion of his thesis research on carotene in butter by chromatogra-phy, stayed on at the University of Missouri with additional chromatographic studies ofnutritional problems, and examined the pigments in other biological tissues: body fat,corpeus luteum and skin secretions of the cow, the yellow pigments in blood serum,the fate of carotenes during digestion and in human milk fat. Palmer extended hisinvestigations to other animals: hen (and eggs), sheep, goat, swine, and horse [1,3], andfound that sheep, swine and rabbit differ from the others in the absence of carotenoidsin their fatty tissue (5 papers in 19151916) [1]. Thus Palmer was the first to introduceadsorption chromatography into the study of animal systems, animal nutrition andbiochemistry.

In 1918, C.H. Eckles moved to the University of Minnesota and invited L.S. Palmer tojoin him. With this transfer, Palmer switched his research area to minerals and vitaminsin animal nutrition and over the subsequent years developed a strong leadership role inthis area regionally (Head of the Division of Agricultural Chemistry, 19421944) andinternationally [1]. However, before leaving carotenoids and chromatography, he wrote athorough 1922 book, Carotinoids and Related Pigments The Chromolipids [6]in which he elaborately detailed the chromatography method and its applications, listedTswetts 13 earlier papers, including a 17 page bibliography, and a detailed discussionof chromatography for laboratory investigations.

The Palmer 1922 book was part of a new monograph series of the American ChemicalSociety and hence received worldwide attention. Bearing in mind, the criticisms of andthe then somewhat obscurity of Tswetts work, Palmers book brought chromatography,carotenoids and Tswetts contributions to scientists elsewhere. Thus Palmers researchand writings serve as the bridge (connection) between Tswett and the resurgence ofchromatography in the early 1930s by R. Kuhn, E. Lederer, P. Karrer and others.(continued in Chapter S-9B, see R. Kuhn.)

See Chapter 5B, b

Other early followers of M.S. Tswett: Gottfried Kranzlin (1882), the first follower of M.S. Tswett, was a botany grad-

uate student at the University of Berlin (19061907). He followed Tswetts 1906description of chromatography, and used a CaCO3 adsorption column to purify

The Beginnings of Chromatography The Pioneers (19001960) 9

chlorophylls and xanthophylls. His 1908 thesis was published in a specialized journaland was soon forgotten. Soon, thereafter, he went to Africa and left the area ofchromatography [1].

Theodor Lippmaa (18921944), an Estonian scientist, received his Ph.D. in botanyin 1926 from Tartu University and published six research papers in 1926 onthuyorhodine, later renamed rhodoxanthin, a plant pigment. For two of thesepapers, he followed Tswetts chromatographic procedures despite the criticism ofTswetts work in the 1920s. His subsequent research was unrelated botanical studiesin Africa and Europe that were a precursor to present-day environmental protection[2].

Charles Dhere (18761955) was the first scientist in Europe to recognize theoverall importance of chromatography [3]. Dhere studied medicine in Paris (M.D.in 1898), but never practiced as a physician. After several years at the Sorbonne,he joined the University of Fribourg, Switzerland in 1900 as an Associate Professorof physiology, biological chemistry and microbiology (such joint appointments werecommon in this period), and then in 1908 as a full Professor until his 1938retirement. His service at Fribourg included two periods as Dean of the Faculty ofScience (19161917 and 19331934). His primary interest was the investigation ofbiological substances, mainly by ultraviolet and fluorescence spectroscopy. One ofDheres students, Wladyslaw Franciszek de Rogowski (18861945) from Poland,received his doctorate in 1912, examined the chromatography on CaCO3 columnsof chlorophylls and their UV absorption and confirmed the earlier findings of M.S.Tswett. Another Dhere student, Guglielmo Vegezzi (18901955) from Switzerland,started his thesis work in 1913, and after military service, completed his doctoralwork in 1916. He extended Rogowskis methods with minor modifications to studyinvertebrate pigments, such as those from bile and liver of escargot and the eggs ofthe spider crab. His research with Dhere is summarized in six papers (19161917).Then he joined the Swiss Federal Administration of Alcohol. Dhere prepared the firstthorough summary of M.S. Tswetts life and scientific work a 50 page paper inthe journal, Candollea. Apparently, Dhere, Rogowski and Vegezzi did not undertakefurther investigations in chromatography after the period described.Thus to evaluate, L.S. Palmer conducted chromatography research from 1910 to

1918, wrote nine papers on carotenoids, and most importantly, wrote a rather thorough1922 chromatography book [6]. Palmer cited Goppelsroeders research, but not that ofKranzlin, Dhere, Rogowski and Vegezzi in his 16 pages of references in 2 columns inhis book. (Scientific communication was more limited at this period in the absence ofairplanes for travel to research conferences, radio, TV, the Web and the Internet.) Tosummarize, L.S. Palmer in the United States and Charles Dhere in Europe were themiddle men, the conservers of Tswetts insights and the bridge to the research of EdgarLederer and Richard Kuhn some 20 years later.

For adsorption chromatography, the next major event came in the laboratory ofRichard Kuhn who had a long, distinguished record of research. One of his researchassistants, Edgar Lederer, had read the above book by Leroy S. Palmer (about 1930)and soon thereafter found the book and papers by Michael Tswett. The other part ofthis story, described under R. Kuhn, is that Kuhn, Winterstein and Lederer published

10 Chapter 1

a key 1931 paper [4] on the purification of xanthophylls on a CaCO3 adsorptionchromatography column. Later, after many other discoveries, R. Kuhn won the NobelPrize in 1938 for his work on carotenoids and vitamins, which will be continuedlater.

Most readers know that Edgar Lederer (19081988) moved in 1933 to France withhis French wife due to political reasons and to the rise of anti-semitism and began anactive research career, mainly at the Institut de Chimie des Substances Naturelles duC.N.R.S. [5,6], and received recognition by being one of the five 1976 M.S. TswettAwardees. Lederers research continued with contributions on carotenoids (astracenefrom red boiled lobster shells, astoxanthine from the skin of goldfish), vitamin A2,perfumes, the pheromone from the queen bee, lysopine from crown gall, ascaryl alcoholfrom a parasitic nematode; microbial lipids, glycolipids and a peptidolipid and muramylpeptides usually with reliance on chromatography. Perhaps more important in thelong run are the many significant chromatography books that he wrote (1934, 1949,1952, 19541957, 1960). This editor read the latter books as a graduate student andremembers the clarity in his writing; these books were probably read by many otheryoung investigators in the 1950s and 1960s. Appendix 3 has several short biographicalarticles about E. Lederer, but his longer, historical biography is lucid and grippingto read for his discoveries, associations and the unfolding of chromatography [6].Lederers shadow is thus long in its present-day influence.

The many transformations of chromatography led some to overlook its quite modestorigins. Indeed, when Kuhn, Winterstein and Lederer presented their chromatographicprogress in the early 1930s at a colloquium at the Chemical Institute of the Universityof Munich, whose Institute Director was the then well-known Henrich Wieland (1928Nobel Prize in Chemistry for his investigations on bile acids) remarked, Up to now,we have learned with much effort to distil, crystallize and recrystallize. Now they comealong and just pour the stuff through a little tube.

See Chapter 5B, b

During the early and mid-1930s, Paul Karrer (18891971) was very active innatural products research at the University of Zurich, Switzerland. After learning theKuhn=Lederer results, he used adsorption chromatography in his investigations andpublished the results in his many research papers in the 1930s. He was recognized as the1937 recipient of the Nobel Prize in Chemistry for his investigations on carotenoids,flavins and vitamins A and B2.

Laszlo Zechmeister (18891972) was another early leader in chromatography. Hewas born in Hungary and was a graduate student of Richard Willstatter before WorldWar I. After the war ended, he returned to his home in Budapest, Hungary and becamein 1923 a Professor at the University of Pecs, Hungary. Being interested in carotenoids,he followed the Kuhn=Lederer group in carotenoid chromatography. In 1934, he wrote abook on carotenoids and then in 1937 wrote the first textbook on chromatography, whichhad three later editions plus his 1950 book, Progress in Chromatography (see A-4A).With the political instability in Europe in the 1930s, he moved to the California Instituteof Technology as a Professor in 1940, and organized a vigorous laboratory to examine

The Beginnings of Chromatography The Pioneers (19001960) 11

natural products by chromatography. Though he retired in 1959, his lucid books werevery effective in the subsequent development and spread of chromatography [7,8].

See Chapter 5B, b

Most scientists remember A.W.K. Tiselius and associate him with free-flow elec-trophoresis, but his fertile mind led also to significant developments in chromatographyin the 1940s and 1950s as described by one of his students, Per Flodin in S-9A.

Aloysius I.M. Keulemans (19081977) was another prominent chromatographer.Most of his career was at the University of Technology, Eindhoven, The Netherlands,where he conducted research on gas chromatography and developed a significantprogram to guide the next generation of chromatographers; this included over 200research papers plus 150 M.S. and 20 Ph.D. degrees for students from his institute.The 1957 and 1959 editions of his textbook on gas chromatography described reportsfrom chromatographers (J.J. Van Deemter, F.J. Zuiderweg, A. Klinkenberg and H. Boerat Shell Development, Amsterdam, The Netherlands) and E. Glueckauf and N.H. Ray(England) and other early investigators [9,10]. The Keulemans books were translated toa number of different languages and served as a major source of information to practicalchromatographers (cited in A-4E).

See Chapter 5B, b

Adsorption chromatography may be extended to become a preparative liquid chro-matography [11,12]. This introduction for adsorption chromatography, whether liquid orgas chromatography, may be supplemented by other sources [13,14]. At this point, anoverall look on the developing patterns for the varieties of chromatography is presented(Fig. 1.1).

Note: Additional comments and references may be found later in Chapter S-9B (R.Kuhn, P. Karrer) and A-4A, A-4E and A-5Ga.

C. PARTITION CHROMATOGRAPHY (1940s1950s)

This subject area has been divided into liquidliquid chromatography and gasliquidchromatography. In an earlier decade, these groups based on the physical state of themobile phase were thought to be distinct; now we know that while one mechanism maybe dominant, other factors are definitely involved.

C.1. Liquidliquid partition chromatography (LLC)

The research of the 1930s, as just described, was dominated by the emphasis onadsorption chromatography. A major turning point came in 1941 with the research byArcher J.P. Martin and Richard L.M. Synge at Cambridge University, United Kingdom[14] on partition chromatography. They used a silica gel column with water as the

12 Chapter 1

Fig. 1.1. Outline of the historical flow of scientific thought in chromatography (1900-1960s). This figurewill serve as an outline for subsequent sections. Partition chromatography and its sequential developmentoccurred over the 1940s to 1960s period. Note: Additional comments and references may be found in laterChapters S-9A, S-9B (R. Kuhn, R. Karrer), A-4A, A-4E and A-5G.

stationary phase plus a mobile phase of water-saturated butanol to separate acetylamino acids to initiate partition chromatography. Column partition chromatography [5]led soon to paper chromatography and thin-layer chromatography, gasliquid partitionchromatography and in the mid-1960s to high-performance liquid chromatography(HPLC).

Note: Further development of TLC is described in Chapters 1-D2, 2, 5 and S-11.

C.2. Gasliquid partition chromatogr