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High Throughput LC-MS/MS Method for Simultaneous Estimation of 9-Cis-Retinoic Acid and its Metabolite 4-Oxo-9-Cis-Retinoic Acid in Human Plasma and its Application to a Bioequivalence Study | Abstract
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
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Research Article

High Throughput LC-MS/MS Method for Simultaneous Estimation of 9-Cis-Retinoic Acid and its Metabolite 4-Oxo-9-Cis-Retinoic Acid in Human Plasma and its Application to a Bioequivalence Study

Arabinda Saha*, Poonam Vats, Richa Thakur, Arshad Khuroo and Tausif Monif

Department of Clinical Pharmacology and Pharmacokinetics, Sun Pharmaceutical Industries Ltd., Haryana, India

*Corresponding Author:
Arabinda Saha
Department of Clinical Pharmacology and Pharmacokinetics
Sun Pharmaceutical Industries Ltd., GP-5, HSIDC
Old Delhi Gurgaon Road, Udyog Vihar Industrial Area
Gurgaon-122 015, Haryana, India
Tel: +91-22-43244324
E-mail: [email protected]

Received date: October 07, 2015; Accepted date: October 26, 2015; Published date: November 02, 2015

Citation: Saha A, Vats P, Thakur R, Khuroo A, Monif T (2015) High Throughput LC-MS/MS Method for Simultaneous Estimation of 9-Cis-Retinoic Acid and its Metabolite 4-Oxo-9-Cis-Retinoic Acid in Human Plasma and its Application to a Bioequivalence Study. J Anal Bioanal Tech S13:001. doi:10.4172/2155-9872.S13-001

Copyright: © 2015 Saha A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

A liquid chromatography mass-spectrometric method is described for the simultaneous estimation of 9-cisretinoic acid and 4-oxo-9-cis-retinoic acid in human plasma using 13-cis-retinoic acid-d5 as an internal standard. Chromatographic separation of these retinoids was achieved on Chromolith Performance RP18 (100 × 4.6 mm) column using binary flow program of two mobile phase. 9-cis-retinoic acid and 4-oxo-9-cis-retinoic acid were extracted from human plasma by liquid-liquid extraction with a mixture of ethyl acetate, n-hexane and iso propyl alcohol (30:65:5, v/v/v). Quantification was achieved by monitoring transitions of m/z 299.4→255.2 for 9-cisretinoic acid, m/z 313.4→269.3 for 4-oxo-9-cis-retinoic acid and 304.4→260.2 for isotretinoin-d5 in multiple reaction monitoring, using turbo ion source in negative polarity. The linearity was established over a dynamic range of 2.85- 450.51 ng/mL and 2.33-180.57 ng/mL for 9-cis-retinoic acid and 4-oxo-9-cis-retinoic acid, respectively. At the lower limit of quantification level, the intra-and inter-day precision values of 9-cis-retinoic acid were within 5.9% and 6.0%, respectively, whereas that of 4-oxo-9-cis-retinoic acid were within 13.8% and 11.3%, respectively. The method was successfully applied for the simultaneous estimation of 9-cis-retinoic acid and 4-oxo-9-cis-retinoic acid in healthy volunteers, after oral administration of 30 mg alitretinoin capsule for bioequivalence study.

Keywords

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Citations : 3732

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