THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 277, No. 40, Issue of October 4, p.

e28, 2002
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A.

Classics
A PAPER IN A SERIES REPRINTED TO CELEBRATE THE CENTENARY OF THE JBC IN 2005

JBC Centennial
1905–2005
100 Years of Biochemistry and Molecular Biology

The Use of Chromatography in Biochemistry

Carotene. VIII. Separation of Carotenes by Adsorption
(Strain, H. H. (1934) J. Biol. Chem. 105, 523–535)
Harold H. Strain received his Ph.D. from Stanford University and worked most of his career
at the Carnegie Institution of Washington, Plant Biology Division, located on the Stanford
campus. He was hired as an organic chemist specifically to aid in the characterization of
pigments from photosynthetic organisms. In the course of this work, he developed a procedure
for the separation, isolation, and characterization of carotenes by adsorption chromatography.

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Adsorption chromatography was first described in 1906 by the Russian botanist Michael
Tswett (1) who successfully fractionated petroleum ether extracts of chlorophyll and other
plant pigments on narrow glass columns packed with dry calcium carbonate. Many of the
leading chemists at the time did not value Tswett’s chromatographic method and made little
use of it. However, several chemists gradually appreciated its value, and by the 1930s
adsorption chromatography was widely used. Strain’s Journal of Biological Chemistry (JBC)
Classic is an excellent example of how organic chemists applied the principles of adsorption
chromatography first enunciated by Tswett for the separation of complex mixtures. The power
of adsorption chromatography must have been recognized by those who subsequently went on
to develop different kinds of chromatographic methods that depend on other principles for
separation including partition chromatography, ion exchange chromatography, gas chroma-
tography, and gel filtration. These methods provided the tools needed by biochemists in the
last half of the twentieth century to separate and characterize the complex mixtures of
compounds in living things that were present in very small amounts, had very similar
properties, were often extremely labile, and were not amenable to separation by the conven-
tional methods of the organic chemist.
In this JBC Classic, Strain explores the use of various adsorbents for the purification of
individual carotenes from mixtures. There was motivation for a new approach because many
of the traditional chemical methods for carotene purification altered them irreversibly, calling
into question the structures of the natural material. Strain tested various adsorbents includ-
ing metallic oxides, charcoal, and fuller’s earth. After preliminary testing, he concluded that
magnesium oxide had the most desirable properties for separation of ␣- and ␤-carotenes and
that the adsorbent did not chemically alter the carotenes being separated. The experimental
description is as useful today as it was in 1934. He reports that using a glass column to hold
the adsorbent was more effective than batch adsorption. He describes packing the columns and
the use of siliceous earth mixed with the adsorbent to increase the flow rate. He also describes
differential elution with solvents of different polarity. Ultimately, he proves that the carotenes
purified by these methods were pure and unaltered by the purification technique.1
Robert D. Simoni, Robert L. Hill, and Martha Vaughan

REFERENCES
1. Tswett, M. S. (1906) Physikalisch-chemische studien uber das chlorophyll. Die adsorptionen. Ber. Bot. Ges. 24,
316 –332

1
We thank Dr. Arthur Grossman, Carnegie Institute for Plant Biology at Stanford University, where Strain did his
work, and Pat Craig for providing biographical information and correspondence that she has collected about Strain for
a history of the Carnegie Institution that she has written.

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 The Use of Chromatography in Biochemistry
Robert D. Simoni, Robert L. Hill and Martha Vaughan
J. Biol. Chem. 2002, 277:e28.

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