EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Quantitative high-performance liquid chromatographic determination of retinoids in human serum using on-line solid-phase extraction and column switching: Determination of 9-cis-retinoic acid, 13-cis-retinoic acid, all-trans-retinoic acid, 4-oxo-all-trans-retinoic acid and 4-oxo-13-cis-retinoic acid



Quantitative high-performance liquid chromatographic determination of retinoids in human serum using on-line solid-phase extraction and column switching: Determination of 9-cis-retinoic acid, 13-cis-retinoic acid, all-trans-retinoic acid, 4-oxo-all-trans-retinoic acid and 4-oxo-13-cis-retinoic acid



Journal of Chromatography B 691(1): 43-58



A fully automated isocratic high-performance liquid chromatographic method for the determination of 9-cis-retinoic acid, 13-cis-retinoic acid, all-trans-retinoic acid, 4-oxo-13-cis-retinoic acid and 4-oxo-all-trans-retinoic acid, has been developed using on-line solid-phase extraction and a column switching technique allowing clean-up and pre-concentration in a single step. A 500-microliter sample of serum was diluted with 750 microliters of a solution containing 20% acetonitrile and the internal standard 9,10-dimethylanthracene. About 1000 microliters of this mixture was injected on a 20 x 4.6 mm I.D. poly ether ether ketone (PEEK) pre-column with titanium frits packed with Bondapak C18, 37-53 microns, 300 A particles. Proteins and very polar compounds were washed out to waste, from the pre-column, with 0.05% trifluoroacetic acid (TFA)-acetonitrile (8.5:1.5, v/v). More than 200 aliquots of diluted serum could be injected on this pre-column before elevated back-pressure enforces replacement. Components retained on the pre-column were backflushed to the analytical column for separation and detection at 360 nm. Baseline separation was achieved using a single 250 x 4.6 mm I.D. Suplex pKb-100 column and a mobile phase containing 69:10:2:16:3 (v/v) of acetonitrile-methanol-n-butanol-2% ammonium acetate-glacial acetic acid. A total time of analysis of less than 30 min, including sample preparation, was achieved. Recoveries were in the range of 79-86%. The limit of detection was 1-7 ng/ml serum and the precision, in the concentration range 20-1000 ng/ml, was between 1.3 and 4.5% for all five compounds. The method was applied for the analysis of human serum after oral administration of 60 mg Roaccutan. The method is well suited for pharmacological studies, while the endogenous levels of some retinoic acid isomers are below the limit of quantitation.

(PDF emailed within 0-6 h: $19.90)

Accession: 009287848

Download citation: RISBibTeXText

PMID: 9140757

DOI: 10.1016/s0378-4347(96)00434-3



Related references

Determination of 13 cis retinoic acid and all trans retinoic acid and aromatic retinoic acid analogs possessing anti tumor activity in biological fluids by high performance liquid chromatography. Journal of Chromatography 152(2): 421-430, 1978

Determination of endogenous levels of 13-cis-retinoic acid , all-trans-retinoic acid and their 4-oxo metabolites in human and animal plasma by high-performance liquid chromatography with automated column switching and ultraviolet detection. Journal of Chromatography B 700(1-2): 31-47, Oct 24, 1997

Determination of endogenous levels of 13-cis-retinoic acid (isotretinoin), all-trans-retinoic acid (tretinoin) and their 4-oxo metabolites in human and animal plasma by high-performance liquid chromatography with automated column switching and ultraviolet detection. Journal of Chromatography, B Biomedical Sciences and Applications 700(1/2): 31-47, 1997

Binding of 9-cis-retinoic acid and all-trans-retinoic acid to retinoic acid receptors alpha, beta, and gamma. Retinoic acid receptor gamma binds all-trans-retinoic acid preferentially over 9-cis-retinoic acid. Journal of Biological Chemistry 269(24): 16689-16695, 1994

The high sensitivity of the rabbit to the teratogenic effects of 13-cis-retinoic acid (isotretinoin) is a consequence of prolonged exposure of the embryo to 13-cis-retinoic acid and 13-cis-4-oxo-retinoic acid, and not of isomerization to all-trans-retinoic acid. Archives of Toxicology 68(2): 119-128, 1994

High pressure liquid chromatographic determination of 13 cis retinoic acid and all trans retinoic acid in human plasma. Analytical Biochemistry 86(2): 743-750, 1978

Novel retinoic acid, 9-CIS retinoic acid, in combination with all-trans retinoic acid is effective inducer of differentiation of retinoic acid resistant Hl 60 cells Relation to retinoic acid receptor and retinoid X receptor expression. Blood 82(10 SUPPL 1): 251A, 1993

Binding of 9-cis-retinoic acid and all-trans-retinoic acid to retinoic acid receptors a, b, and c. Retinoic acid receptor c binds all-trans-retinoic acid preferentially over 9-cis-retinoic acid. The Journal of Biological Chemistry 269: 689-95, 1994

Novel retinoic acid, 9-cis retinoic acid, in combination with all-trans retinoic acid is an effective inducer of differentiation of retinoic acid-resistant HL-60 cells. Blood 83(11): 3289-3297, 1994

Determination of retinoic acid (13-cis- and all-trans-) and aromatic retinoic acid analogs possessing anti-tumor activity, in biological fluids by high-performance liquid chromatography. Journal of Chromatography 152(2): 421-430, 1978

Improved and rapid high-performance liquid chromatographic assay for 13-cis-retinoic acid or all-trans-retinoic acid. Journal of Pharmaceutical Sciences 71(2): 262-264, 1982

Quantitative axial profiles of retinoic acid in the embryonic mouse spinal cord: 9-cis retinoic acid only detected after all-trans-retinoic acid levels are super-elevated experimentally. Developmental Dynamics 222(3): 341-353, 2001

Correlations between conceptal concentrations of all trans retinoic acid and dysmorphogenesis after microinjections of all trans retinoic acid 13 cis retinoic acid all trans retinoyl beta glucuronide or retinol in cultured whole rat embryos. Drug Metabolism & Disposition 20(2): 218-225, 1992

A sensitive high-pressure liquid chromatography/particle beam/mass spectrometry assay for the determination of all-trans-retinoic acid and 13-cis-retinoic acid in human plasma. Journal of Pharmaceutical Sciences 85(3): 287-290, 1996

The high sensitivity of the rabbit to the teratogenic effects of 13-cis-retinoic acid is a consequence of prolonged exposure of the embryo to 13-cis-retinoic acid, and not of isomerization to all-trans retinoic acid. Teratology 47(5): 438, 1993