Journal of Lipid Research, Vol 25, 400-406, Copyright © 1984 by Lipid Research, Inc.
Separation and quantitation of serum beta-carotene and other carotenoids by high performance liquid chromatography
N Katrangi, LA Kaplan and EA Stein
We describe a specific assay for serum provitamin A (alpha- and beta-
carotene) by high pressure liquid chromatography (HPLC). The system
separates alpha- and beta-carotene in 7.4 min using a C18 muBondapak,
10-micron particle size column with a mobile phase of acetonitrile-
chloroform 92:8 at 2 ml per min and a 462 nm detector. The HPLC assay had a
recovery of 94.8% of added beta-carotene and, at a serum concentration of
215.2 micrograms/L, had within-run and between-run precisions of 3.1% and
3.6%, respectively. In 65 subjects, the HPLC- determined provitamin A
(alpha- and beta-carotene) value was 343 +/- 166 micrograms/L and averaged
23.4 +/- 7.9% (range 9-43%) of the values obtained by a traditional
colorimetric assay for total serum "carotenes." Although total serum
carotenes showed no relationship to serum vitamin A (r = -0.048; P = 0.78),
HPLC-determined alpha- and beta- carotene was significantly inversely
correlated (r = -0.357; P = 0.05).