SIMULTANEOUS DETERMINATION OF ALL-TRANS-RETINOIC, 13-CIS-RETINOIC AND9-CIS-RETINOIC ACID, THEIR 4-OXO METABOLITES AND ALL-TRANS-RETINOL INHUMAN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
C. Lanvers et al., SIMULTANEOUS DETERMINATION OF ALL-TRANS-RETINOIC, 13-CIS-RETINOIC AND9-CIS-RETINOIC ACID, THEIR 4-OXO METABOLITES AND ALL-TRANS-RETINOL INHUMAN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, Journal of chromatography B. Biomedical applications, 685(2), 1996, pp. 233-240
Citations number
25
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Journal title
Journal of chromatography B. Biomedical applications
All-trans-retinoic acid (all-trans-RA) and 13-cis-retinoic acid (13-ci
s-RA), due to their effects on cell differentiation, proliferation and
angiogenesis, improved treatment results in some malignancies. Pharma
cokinetic studies of all-trans-RA and 13-cis-RA along with monitoring
of retinoic acid metabolites may help to optimize retinoic acid therap
y and to develop new effective strategies for the use of retinoic acid
s in cancer treatment. Therefore, we developed a HPLC method for the s
imultaneous determination in human plasma of the physiologically impor
tant retinoic acid isomers, all-trans-, 13-cis- and 9-cis-retinoic aci
d, their 4-oxo metabolites, 13-cis-4-oxoretinoic acid (13-cis-4-oxo-RA
) and all-trans-4-oxoretinoic acid (all-trans-4-oxo-RA), and vitamin A
(all-trans-retinol). Analysis was performed on a silica gel column wi
th UV detection at 350 nm using a binary multistep gradient composed o
f n-hexane, 2-propanol and glacial acetic acid. For liquid-liquid extr
action a mixture of n-hexane, dichloromethane and 2-propanol was used.
The limits of detection were 0.5 ng/ml for retinoic acids and 10 ng/m
l for all-trans-retinol. The method showed good reproducibility for al
l components (within-day C.V.: 3.02-11.70%; day-to-day C.V.: 0.01-11.3
4%). Furthermore, 9-cis-4-oxoretinoic acid (9-cis-4-oxo-RA) is separat
ed from all-trans-4-oxo-RA and 13-cis-4-oxo-RA. In case of clinical us
e of 9-cis-retinoic acid (9-cis-RA) the pharmacokinetics and metabolis
m of this retinoic acid isomer can also be examined.