History of chromatography

The history of chromatography spans from the mid-19th century to the 21st. Chromatography, literally "color writing", was used—and named— in the first decade of the 20th century, primarily for the separation of plant pigments such as chlorophyll (which is green) and carotenoids (which are orange and yellow). New forms of chromatography developed in the 1930s and 1940s made the technique useful for a wide range of separation processes and chemical analysis tasks, especially in biochemistry.

Precursors
The earliest use of chromatography—passing a mixture through an inert material to create separation of the solution components based on differential adsorption—is sometimes attributed to German chemist Friedlieb Ferdinand Runge, who in 1855 described the use of paper to analyze dyes. Runge dropped spots of different inorganic chemicals onto circles of filter paper already impregnated with another chemical, and reactions between the different chemicals created unique color patterns. According to historical analysis of L. S. Ettre, however, Runge's work had "nothing to do with chromatography" (and instead should be considered a precursor of chemical spot tests such as the Schiff test).

In the 1860s, Christian Friedrich Schönbein and his student Friedrich Goppelsroeder published the first attempts to study the different rates at which different substances move through filter paper. Schönbein, who thought capillary action (rather than adsorption) was responsible for the movement, called the technique capillary analysis, and Goppelsroeder spent much of his career using capillary analysis to test the movement rates of a wide variety of substances. Unlike modern paper chromatography, capillary analysis used reservoirs of the substance being analyzed, creating overlapping zones of the solution components rather than separate points or bands. However, in his book Capillaranalyse ...  (1901), Goppelsroeder stated (p. 168) that he had been separating plant colorants since 1880, and that he had achieved complete separations of those colorants. From p. 166: ''"Bietet sich auch dem Auge bei Betrachtung der verschiedenen Pflanzenorgane eine wunderbare Mannigfaltigkeit der Farben und Farbenabstufungen dar, so bleibt ihm doch die wichtige Thatsache verborgen, dass meist nicht nur ein einziger Farbstoff, sondern mehrere nebeneinander in demselben Organe vorkommen. Während das Auge nur eine Färbung erkennt und wir desshalb glauben, dass dieselbe einem bestimmten einzelnen Farbstoff angehöre, lässt uns die Capillaranalyse meist mehrere verschieden gefärbte Zonen auf den Capillarstreifen in bestimmer, sehr oft von farblosen Zonen unterbrochener Reihenfolge erkennen.  Das Chlorophyll oder Blattgrün z.B. findet sich nicht nur in den grünen, sonder auch in anders gefärbten Organen, beispielsweise verdeckt durch die rote Färbung des Zellsafts in den Blättern der Blutbuche neben dem roten Anthokyan, sowie neben roten Phycoerythrin in den Rotalgen, den Florideen. Diese verschiedenen Farbstoffe lassen sich durch Capillaranalyse in den gemeinschaftlichen Auszügen, ohne irgend welche sonstige Trennungsmanipulationen nebeneinander nachweisen. Sind sie capillarisch in Zonen getrennt, dann genügt deren spectroscopische und chemische Prüfung zur endgiltigen Feststellung ihrer Natur." (If a wonderful variety of colors and color gradations presents itself to the eye when looking at the different plant organs, yet the important fact remains hidden to it: that usually not only a single colorant but several occur side by side in the same organs. While the eye perceives only one color and we believe therefore that it belongs to a certain individual colorant, capillary analysis [i.e., paper chromatography] allows us to detect usually several differently colored zones on the capillary strips in certain sequences [that are] very often interrupted by colorless zones.  Chlorophyll or leaf green, for example, is found not only in the green, but also in differently colored organs; for example, obscured by the red color of the protoplasm in the leaves of the copper beech together with red anthocyanin, as well as together with red phycoerythrin in red algae, the Florideae''. These various colorants can be detected by capillary analysis in extracts where they are present in combination, without any other concurrent separation treatments. If they are separated into zones by capillarity, then their spectroscopic and chemical examination suffices for the conclusive ascertaining of their nature.)

Work on capillary analysis continued, but without much technical development, well into the 20th century. The first significant advances over Goppelsroeder's methods came with the work of Raphael E. Liesegang: in 1927, he placed filter strips in closed containers with atmospheres saturated by solvents, and in 1943 he began using discrete spots of sample adsorbed to filter paper, dipped in pure solvent to achieve separation. This method, essentially identical to modern paper chromatography, was published just before the independent—and far more influential—work of Archer Martin and his collaborators that inaugurated the widespread use of paper chromatography.

In 1897, the American chemist David Talbot Day (1859–1915), then serving with the U.S. Geological Survey, observed that crude petroleum generated bands of color as it seeped upwards through finely divided clay or limestone. In 1900, he reported his findings at the First International Petroleum Congress in Paris, where they created a sensation. Soon after David T. Day's discovery, other researchers investigated the diffusion of petroleum through finely divided earths; viz, the German organic chemist Karl Engler (1842–1925) of the Technical University Karlsruhe and the American chemist Joseph Elliot Gilpin (1866–1924) of Johns Hopkins University:
 * Engler, Karl Oswald Viktor and von Heimhalt, Hans Höfer (1913) Das Erdöl: seine Physik, Chemie, Geologie, Technologie und Wirtschaftsbetrieb (Petroleum: Its physics, chemistry, geology, technology and commercial operation), vol. 1. Leipzig, Germany: S. Hirzel; p. 130:  "Die Dayschen Versuche erregten damals Aufsehen, da er sie in Beziehung zur Genesis der Erdöle brachte, ... "  (Day's experiments created a sensation at that time, since he related them [i.e., his experiments] to the creation of petroleum, ... )
 * Reprinted in and in
 * Reprinted in
 * Engler, Karl Oswald Viktor and von Heimhalt, Hans Höfer (1913) Das Erdöl: seine Physik, Chemie, Geologie, Technologie und Wirtschaftsbetrieb (Petroleum: Its physics, chemistry, geology, technology and commercial operation), vol. 1. Leipzig, Germany: S. Hirzel; p. 130:  "Die Dayschen Versuche erregten damals Aufsehen, da er sie in Beziehung zur Genesis der Erdöle brachte, ... "  (Day's experiments created a sensation at that time, since he related them [i.e., his experiments] to the creation of petroleum, ... )
 * Reprinted in and in
 * Reprinted in

Tsvet and column chromatography
The first true chromatography is usually attributed to the Russian-Italian botanist Mikhail Tsvet. Tsvet applied his observations with filter paper extraction to the new methods of column fractionation that had been developed in the 1890s for separating the components of petroleum. He used a liquid-adsorption column containing calcium carbonate to separate yellow, orange, and green plant pigments (what are known today as xanthophylls, carotenes, and chlorophylls, respectively). The method was described on December 30, 1901, at the 11th Congress of Naturalists and Doctors (XI съезд естествоиспытателей и врачей) in Saint Petersburg. The first printed description was in 1903, in the Proceedings of the Warsaw Society of Naturalists, section of biology. He first used the term chromatography in print in 1906 in his two papers about chlorophyll in the German botanical journal, Berichte der Deutschen Botanischen Gesellschaft. In 1907 he demonstrated his chromatograph for the German Botanical Society. Mikhail's surname "Цвет" means "color" in Russian, so there is the possibility that his naming the procedure chromatography (literally "color writing") was a way that he could make sure that he, a commoner in Tsarist Russia, could be immortalized.

In a 1903 lecture (published in 1905), Tsvet also described using filter paper to approximate the properties of living plant fibers in his experiments on plant pigments—a precursor to paper chromatography. He found that he could extract some pigments (such as orange carotenes and yellow xanthophylls) from leaves with non-polar solvents, but others (such as chlorophyll) required polar solvents. He reasoned that chlorophyll was held to the plant tissue by adsorption, and that stronger solvents were necessary to overcome the adsorption. To test this, he applied dissolved pigments to filter paper, allowed the solvent to evaporate, then applied different solvents to see which could extract the pigments from the filter paper. He found the same pattern as from leaf extractions: carotene could be extracted from filter paper using non-polar solvents, but chlorophyll required polar solvents.

Tsvet's work saw little use until the 1930s.

Martin and Synge and partition chromatography
Chromatography methods changed little after Tsvet's work until the explosion of mid-20th century research in new techniques, particularly thanks to the work of Archer John Porter Martin and Richard Laurence Millington Synge. By "the marrying of two techniques, that of chromatography and that of countercurrent solvent extraction", Martin and Synge developed partition chromatography to separate chemicals with only slight differences in partition coefficients between two liquid solvents. Martin, who had previously been working in vitamin chemistry (including attempts to purify vitamin E), began collaborating with Synge in 1938, brought his experience with equipment design to Synge's project of separating amino acids. After unsuccessful experiments with complex countercurrent extraction machines and liquid-liquid chromatography methods where the liquids move in opposite directions, Martin hit on the idea of using silica gel in columns to hold water stationary while an organic solvent flows through the column. Martin and Synge demonstrated the potential of the methods by separating amino acids marked in the column by the addition of methyl red. In a series of publications beginning in 1941, they described increasingly powerful methods of separating amino acids and other organic chemicals.

In pursuit of better and easier methods of identifying the amino acid constituents of peptides, Martin and Synge turned to other chromatography media as well. A short abstract in 1943 followed by a detailed article in 1944 described the use of filter paper as the stationary phase for performing chromatography on amino acids: paper chromatography. By 1947, Martin, Synge and their collaborators had applied this method (along with Fred Sanger's reagent for identifying N-terminal residues) to determine the pentapeptide sequence of Gramicidin S. These and related paper chromatography methods were also foundational to Fred Sanger's effort to determine the amino acid sequence of insulin.

Refining the techniques
Martin, in collaboration with Anthony T. James, went on to develop gas chromatography (GC; the principles of which Martin and Synge had predicted in their landmark 1941 paper) beginning in 1949. In 1952, during his lecture for the Nobel Prize in Chemistry (shared with Synge, for their earlier chromatography work), Martin announced the successful separation of a wide variety of natural compounds by gas chromatography. Previously, Erika Cremer had laid the theoretical basis of GC in 1944 and Austrian chemist Fritz Prior, under the direction of Erika Cremer, constructed in 1947 the first prototype of a gas chromatograph and achieved separating oxygen and carbon dioxide, in 1947 during his Ph.D. research.

The ease and efficiency of gas chromatography for separating organic chemicals spurred the rapid adoption of the method, as well as the rapid development of new detection methods for analyzing the output. The thermal conductivity detector, described in 1954 by N. H. Ray, was the foundation for several other methods: the flame ionization detector was described by J. Harley, W. Nel, and V. Pretorius in 1958, and James Lovelock introduced the electron capture detector that year as well. Others introduced mass spectrometers to gas chromatography in the late 1950s.

The work of Martin and Synge also set the stage for high performance liquid chromatography, suggesting that small sorbent particles and pressure could produce fast liquid chromatography techniques. This became widely practical by the late 1960s (and the method was used to separate amino acids as early as 1960).

Thin layer chromatography
The first developments in thin layer chromatography occurred in the 1940s, and techniques advanced rapidly in the 1950s after the introduction of relatively large plates and relatively stable materials for sorbent layers.

Later developments
In 1987 Pedro Cuatrecasas and Meir Wilchek were awarded the Wolf Prize in Medicine for the invention and development of affinity chromatography and its applications to biomedical sciences.