APPLICATION OF CLOUD-POINT EXTRACTION-REVERSED-PHASE HIGH-PERFORMANCELIQUID-CHROMATOGRAPHY - A PRELIMINARY-STUDY OF THE EXTRACTION AND QUANTIFICATION OF VITAMIN-A AND VITAMIN-E IN HUMAN SERUM AND WHOLE-BLOOD
Sr. Sirimanne et al., APPLICATION OF CLOUD-POINT EXTRACTION-REVERSED-PHASE HIGH-PERFORMANCELIQUID-CHROMATOGRAPHY - A PRELIMINARY-STUDY OF THE EXTRACTION AND QUANTIFICATION OF VITAMIN-A AND VITAMIN-E IN HUMAN SERUM AND WHOLE-BLOOD, Journal of chromatography B. Biomedical sciences and applications, 716(1-2), 1998, pp. 129-137
Citations number
25
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Journal title
Journal of chromatography B. Biomedical sciences and applications
Methods available for quantification of vitamins A and E in serum or b
lood requires preconcentration and clean-up by liquid-liquid extractio
n, evaporation of the extract, and reconstitution of the extract in a
solvent of choice before analysis. This process not only involves the
use of toxic organic solvents but also requires a long sample preparat
ion time. The lipids and other non-polar coextractants often require a
dditional steps for sample clean-up and evaporation, which may cause s
ample losses. The use of cloud-point extraction eliminates most of the
se sample clean-up problems. We recently demonstrated that cloud-point
extraction (CPE) can be used for extraction and quantification of pol
ycyclic aromatic hydrocarbons (PAHs) and polychlorinated dibenzo-p-dio
xins (PCDDs) from human serum. We now demonstrate how CPE can be used
with human serum and blood, at volumes as low as 50 mu l, and report a
methodology for extracting and quantifying two clinically important v
itamins, (A and E) from human serum and blood. Vitamins A and E were e
xtracted from human serum and blood by using Genapol X-80 as the cloud
-point extractant under salting out conditions. Serum and blood sample
s were diluted in organic-free water to get sufficiently large sample
volumes for CPE. The surfactant-rich phases were separated by centrifu
gation, and the samples were analyzed by HPLC-UV after deleterious coe
xtractants were removed by precipitating them with acetonitrile. The r
ecoveries of spiked vitamins A and E were found to be 85.6+/-0.4% and
82.6+/-5.2%, respectively. The average concentration of vitamins A and
E in a serum pool after correction for recoveries were found to be 43
.4+/-1.8 mu g/dl (1.5 +/- 0.1 mu mol/l) and 564.3 +/- 65.3 mu g/dl (13
.1 +/- 1.5 mu mol/l), respectively. Vitamin A and E concentrations in
whole blood were found to be 26.3+/-0.4 mu g/dl (0.92+/-0.01 mu mol/l)
and 457.5+/-15.6 mu g/dl (10.6+/-0.4 mu mol/l), respectively. These v
alues are comparable with those obtained by the reference method used
at the Centers for Disease Control and Prevention. The success of the
preliminary study will lead to a comprehensive validation of this meth
od for vitamins A and E in serum and blood. (C) 1998 Elsevier Science
BN. All rights reserved.