Journal of Analytical & Bioanalytical Techniques
Open Access

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

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: arabinda.saha@sunpharma.com

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.

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.

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