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Clinical Chemistry 40: 48-57, 1994;
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Clinical Chemistry, Vol 40, 48-57, Copyright © 1994 by American Association for Clinical Chemistry

Simultaneous determination of endogenous retinoic acid isomers and retinol in human plasma by isocratic normal-phase HPLC with ultraviolet detection

E Meyer, WE Lambert and AP De Leenheer
Laboratory of Medical Biochemistry and of Clinical Analysis, Faculty of Pharmaceutical Sciences, University of Ghent, Belgium.

We describe a rapid and simple procedure for the simultaneous quantitation of endogenous 13-cis-retinoic acid, all-trans-retinoic acid, and retinol by isocratic normal-phase HPLC with ultraviolet detection, in 0.5 mL of human plasma. A silica adsorption column was eluted with n-hexane:2-propanol:acetic acid (200:0.7:0.135 by vol) at 0.9 mL/min, and the effluent monitored at 350 nm. The arotinoid ethylsulfonic acid Ro 15-1570 was used as the internal standard. High sensitivity, allowing quantitation of physiological concentrations, was achieved, particularly for the retinoic acid isomers. The detection limits were 0.5 microgram/L in plasma for both 13-cis- and trans- retinoic acid, and 10 micrograms/L for retinol. The CVs for between-day determinations of the lowest quality-control concentration (n = 12) were 4.8% for 13-cis-retinoic acid, 3.4% for trans-retinoic acid, and 3.0% for retinol. The mean (+/- SD) concentrations of 13-cis-retinoic acid (1.79 +/- 0.56 microgram/L), trans-retinoic acid (1.35 +/- 0.42 microgram/L), and retinol (533 +/- 58 micrograms/L) measured in plasma from 22 healthy volunteers agreed well with those previously reported.


The following articles in journals at HighWire Press have cited this article:


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 Am. J. Physiol. Endocrinol. Metab.Home page
E. A. Werner and H. F. Deluca
Retinoic acid is detected at relatively high levels in the CNS of adult rats
Am J Physiol Endocrinol Metab, March 1, 2002; 282(3): E672 - E678.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
J. C. White, V. N. Shankar, M. Highland, M. L. Epstein, H. F. DeLuca, and M. Clagett-Dame
Defects in embryonic hindbrain development and fetal resorption resulting from vitamin A deficiency in the rat are prevented by feeding pharmacological levels of all-trans-retinoic acid
PNAS, November 10, 1998; 95(23): 13459 - 13464.
[Abstract] [Full Text] [PDF]


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