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Science Tribune - Article - October 1996

http://www.tribunes.com/tribune/art96/jose.htm

The inventor of chromatography (1861) :
F. Goppelsröder, professor at the University of Basel



P. Jössang

Chemistry Laboratory, Natural History Museum, Paris, France.
(translated from the French by Tiiu Ojasoo)


Browsing in the library

The normally peaceful atmosphere of the Chemistry Laboratory library, one of the oldest in France, was suddenly shaken by an abrupt increase in entropy - arising from somewhere in the Administration. Our wigged ancestors, among them Vauquelin (1763-1829) - who discovered chromium and glucinium oxide (beryllium) - were alone in contemplating with serenity the piles of old books that had been torn away from their ecotope.

As a regular browser, I rummaged among the piles fearing some impending autodafé justified by the 'spirit of the times' which considers with pity any book published before - say -1985. Such an attitude is forgivable in young students but, nevertheless, a serious mistake. 'Paleochemistry' (in particular when relating to the 19th C) is an underexploited treasure trove, very rich in achievements that could well be pursued with our present technical means and knowledge.

I peered into the books, flipped through their pages and, in so doing, came across a Jahresbericht über die Fortschritte der Chemie ... für 1862 (Giessen) ("Annual Review of Progress in Chemistry ......, 1862"), a kind of "Chemical Abstracts" before their day. Tucked between a description of a Dragendorff apparatus for the assay of lipids in some kind of seeds and a paper by Mr. Müller on milk lipids, there were seven lines, no more :

"F. GOPPELSRÖDER (....) has shown
that SCHÖNBEIN's observation - whereby
solutions of various substances are aspired
with very different rates and intensities by
filter paper - can be used to separate and
distinguish different dyes contained in the
solution."

My head swam for this was nothing other than "paper chromatography" for which Martin and Synge had received the Nobel Prize in chemistry in 1952. It was a special type of chromatography called "frontal analysis" that P. Chovin, in the magnificent treatise on 'Chromatography in Organic and Biologic Chemistry' published under the leadership of E. Lederer (1), had attributed to Tiselius (2) and Claesson (3).


Discovery of Goppelsröder's 1861 reference

I did not have access to the references that were cited in connection with Friedrich Goppelsröder's discovery in the'Jahresbericht'. It was only by pure chance that I came across a large tome by Goppelsröder among the piled-up 'old' books in the library: "Studien über die Anwendung der Capillaranalyse....." (" Studies on capillary analysis......") published in 1904 (4). This was the author's key work and was also to prove to be the key to an unexpected discovery !

My attention had been drawn by the title of the book because, at the time, I was looking for an original article on "capillary analysis" to find out what it was all about. (The term "capillary analysis" is recent and, according to Goppelsröder, was coined in 1887). The two primary publications on the subject were dated 1861 :

- "Über ein Verfahren die Farbstoffe in ihren Gemischen zu erkennen". Verhandlungen der Naturforschenden Gesellschaft zu Basel, 1861, III, 2. Heft ("On a procedure for identifying dyes in a mixture thereof". Transactions of the Society for Natural Sciences of Basel, 1861, III, part 2)

- "Note sur une méthode nouvelle propre à déterminer la nature d'un mélange de principes colorants", séance du 30 octobre 1861, Bulletin de la Société Industrielle de Mulhouse, Tome XXXII, 1862 ("Memorandum on a new method for determining the nature of a mixture of dyes", Meeting held on October 30, 1861, Bulletin of the Industrial Society of Mulhouse,Vol XXXII, 1862).

Our library held the Bulletin of the Industrial Society of Mulhouse !


Schönbein's early observations on frontal analysis

In his article published in the Bulletin, Goppelsröder - with highly laudable honesty - makes it clear right from the start that it is "M. Schönbein who presented some time ago, at the Society of Natural Sciences of Basel, highly interesting observations on the different heights to which different substances in solution can rise by infiltrating the pores of Joseph paper (a kind of filter paper)". He describes some of Schönbein's experiments and I shall cite just one which illustrates very well the principle of frontal analysis and of the basic technique that was employed by Schönbein and much developed by Goppelsröder :

"A strip of filter paper (is dipped) several millimeters into an aqueous solution of blue litmus. The solution is seen to rise quickly above the level of the liquid by capillary aspiration. A blue band is formed but, because the water and dye both rise to the same extent, no separation between solvent and dissolved substance is observed. A different situation arises when the experiment is repeated with litmus pinkened with sulfuric acid. In this case, three zones are formed on the paper above the liquid. The first (i.e. the highest) contains only water, the second is due to dilute sulfuric acid, and the third contains water, acid, and dye. Clearly, a partial separation of the three components in the mixture has occurred and is due to their different abilities to rise within the pores of the paper."

Goppelsröder adds the following capital remark: "I saw in these observations the key to a new analytical method" and "with M. Schönbein's permission (...) I started (...) to examine the behaviour of dyes from this aspect."


Goppelsröder invents preparative paper chromatography and chromatographic reagents

Goppelsröder dried the filter paper used in the analysis, cut out a relevant zone, eluted the product with a solvent (e.g. ethanol), and submitted the products obtained to further analysis. In this way, he detected picric acid, a yellow substance that migrates upfront (i.e. upwards) from within a mixture that, in his experiment, was unpurified fuchsin prepared from aniline and in solution in ethanol. Picric acid (2,4,6-trinitrophenol) has been used as an explosive in shells under the name 'Melinite'.

Goppelsröder also isolated pure azulene - a blue hydrocarbon of curious structure that has always fascinated chemists - by repeated capillary analysis (preparative paper chromatography). The pink-coloured impurity was probably fuchsin. He coloured small skeins of silk either in pink with the impurity or in blue with pure azulene. He concludes (may I remind you that we are in 1861 !) :

" I am convinced that this method will prove to be very practical for the rapid determination of the nature of a mixture of dyes, especially if appropriately chosen and characterized REAGENTS are used...."

This is nothing less than the discovery of chromatographic reagents which are essential agents still in daily use in 1996, to detect and characterize colourless products in particular.


Goppelsröder perfects his method

From 1861 onwards, Goppelsröder spent his time improving his method. He describes his results in the introduction to the "Studien über die Anwendung ...." (4) in some detail. What were his key discoveries and applications ?

1) By repeated analysis and band elution, Goppelsröder neatly separated the components in mixtures of dozens of compounds,

2) Goppelsröder detected colourless compounds using chromatographic reagents that yielded information on the nature of these compounds,

3) He used his method to isolate pure compounds,

4) Goppelsröder characterized compounds by spectral methods (emission or absorption) or by microchemical reactions. (In 1996, chromatography and spectrography are methods that are still regularly used in laboratories studying natural substances). In the conclusion of his book (4, p. 153), he mentions direct (absorption) spectrography that does away with the isolation step. The filter paper was steeped in, for example, wintergreen oil which makes it transparent in the spectral zone under study.

5) Goppelsröder used papers impregnated with organic or mineral substances as well as materials other than paper (cotton, linen, wool, and silk) (4, p. 152) or even transformed paper (oxycellulose...)

6) He analyzed an extensive range of compounds (mineral, organic (including macromolecules)). His interests covered the analysis of medical specimens, consumer protection, and forensic science (the detection of picric acid in beer).


Some examples of Goppelsröder's reagents and analyses

Reagents :

- Fehling's solution for detecting reducing sugars,
- 0.1% furfural and sulfuric acid for detecting bile acids,
- Wurster's reagent : in the presence of quinone (traces) and sodium carbonate, the amino-acid leucine gives rise to a violet coulour,
- Millon's reagent for tyrosine, another amino-acid,
- two reagents for albumin,
- a series of reagents (Huppert, Gmelin, Hammarsten) for biliverdine. Goppelsröder often gave the sensitivity of the reagents he used : 1/80 000 for Gmelin's reagent.

Several other reactions could be added to this list : Liebermann's, Biuret, (proteins and urea), xanthoproteic, Jaffé, Legal, the orcin-chlorhydric reaction of pentoses......

Compounds analyzed :

Besides studying dyes, Goppelsröder also extensively analyzed 'biochemical' substances. Apart from those cited above, let's mention hippuric acid, taurocholic acid, urea, uric acid, creatinine, "the" globulin, serum-albumin, fibrin, hemoglobin, hematin, hematoporphyrin, urobilin, bilirubin, indole, scatole.

Much of Goppelsröder's book is devoted to the analysis of 507 urine samples from 178 patients with 86 diseases and is an attempt to establish correlations. Goppelsröder gives very detailed and well illustrated results and, in this respect, can be considered a true pioneer of medical analysis.

He also studied plant substances but I was unable to refer to the cited work (5).


One hundred years ahead of his time

Clearly, Goppelsröder was about 100 years ahead of his time. The techniques he used and perfected are often representative of chromatography as practised in the second half of the 20th c. The way he defined capillary analysis is most interesting (4, p.152) :

"Capillary analysis means that..... compounds dissolved in a solvent are concentrated by capillary and absorption forces into specific zones of a capillary column of products. .."

Let's see how this matches with the defintion of chromatography given in 1944 by the Nobel Prize winners, Martin and Synge (6) :

(Chromatography is a) "technical procedure of analysis by percolation of fluid through a body of comminuted or porous rigid material, irrespective of the nature of the physicochemical processes that may lead to the separation of substances in the apparatus."

Undisputedly, capillary analysis meets this definition.


Nobel prize winners, Tsvett, or Goppelsröder ?

According to Leslie S. Ettre, a well-known expert on the history of chromatography, the first papers on separation by chromatography are by M.S. Tswett (7). What was Tsvett's contribution ? For sure, the systematic use of a chromatography column but, primarily, the use of a narrow zone of mixture to be separated which, in contrast to frontal analysis, is made to migrate across a porous medium by solvents that do not contain the compounds to be separated. Under ideal conditions, this gives rise to "bands" of pure compounds that are separated in a single step. In other words, Tsvett turned chromatography into a handy procedure.

When did Tsvett publish his first papers ? Tsvett himself gives us the answer to this question in a reprint which bears the hand-written mention : "With the compliments of the author" (8):

"The most suitable method for obtaining information on the number and nature of components of chlorophyll seems to be the one I have elaborated over the last ten years under the name adsorption chromatography" (reference published in 1906 (9)).


Epilogue

The reader can come to his own conclusions. I shall just add one last comment. Leslie S. Ettre mentions that Tswett (sic), of Russian origin, was educated in Switzerland and studied at the University of Geneva.

No question ! Switzerland has given birth to chromatography - twice !




References

1. Chovin P. La chromatographie en chimie organique et biologique. Masson, Paris, 1959, II, p. 25.

2. Tiselius A. Arkiv Kemi Mineral Geol 14B, n°22, 1940-41.

3. . Claesson S. Ibid. 20A, n°3, 1945.

4. Goppelsröder F. Studien über die Anwendung der Capillaranalyse..... (" Studies on capillary analysis......"), Birkhauser, Basel, 1904.

5. Goppelsröder F. Verhandlungen der Naturforschenden Gesellschaft zu Basel Vol. XIV, 1901.

6. Gordon AH, Martin AJP, Synge RLM. Biochem JI 38, 65-68, 1944; Consden R, Gordon AH, Martin AJP. Biochem J 38, 224, 1944.

7. Ettre LS. International Laboratory, Sept 1991, p.18.

8. Tsvett MS. L'état actuel de nos connaissances sur la chimie de la chlorophylle. Extrait de la Revue Générale des Sciences num. du 29 février 1912, Librairie Armand Colin, Paris.

9. Tsvett MS. Berichte d. D. Botan. Ges XXIV, p. 316 et 384, 1906 et les Chromophylles (sic) dans les mondes végétal et animal. Warsaw, 1910.



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