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Hindi Research Article
Determination of Ethambutol in Presence of Fixed Dose Combination Molecules from Human Plasma by Direct Injection to Liquid Chromatography Tandem Mass Spectrometry
| Chaitanya Krishna A1*, Saravanan RS2, Sathiyaraj M2, Jeevanantham S2 and Baskaran R2 | |
| 1Study Director, Bombay Bioresearch Centre, Mumbai, India | |
| 2Department of Bioanalaytical, Bombay Bioresearch Centre, Mumbai, India | |
| Corresponding Author : | Chaitanya Krishna A Study Director, Bombay Bioresearch Centre Govandi-400043, Mumbai, India E-mail: chaitanya@bbrc-cro.com |
| Received February 21, 2012; Accepted March 31, 2012; Published April 05, 2012 | |
| Citation: Chaitanya Krishna A, Saravanan RS, Sathiyaraj M, Jeevanantham S, Baskaran R (2012) Determination of Ethambutol in Presence of Fixed Dose Combination Molecules from Human Plasma by Direct Injection to Liquid Chromatography Tandem Mass Spectrometry. Clin Pharmacol Biopharm. 1:101. doi:10.4172/2167-065X.1000101 | |
| Copyright: © 2012 Chaitanya Krishna 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 simple, rapid, specific and sensitive liquid chromatography tandem mass spectrometric method has been developed and validated for the estimation of Ethambutol from 100 μL of human plasma. Ethambutol is extracted from human plasma by Protein Precipitation Extraction. Glipizide was used as an internal standard. Detection was performed using TSQ Quantum Discovery max mass spectrometer with ESI source in positive polarity. The detection transition for Ethambutol is 205.230 → 116.090 and for Glipizide is 446.200 → 321.200. Chromatographic separation of analyte and internal standard were carried out using a reverse phase Agilent, Eclipse XDB-C18, 4.6 X 150 mm, 5 μ at a flow rate of 0.500 mL/min. The mobile phase is composed of methanol: 0.1 %TFA in 5 mM Ammonium Acetate (90:10) v/v. The assay of Ethambutol is linear over the range of 0.106 μg /mL to 7.006 μg/mL with a precision < 9.91% and the limit of quantification in plasma for Ethambutol was 0.106 μg/mL. Mean extraction recovery obtained was 98.70%. Samples are stable at room temperature for 6 hrs, processed samples were stable at least for 28 hrs and also stable at three freeze–thaw cycles. The method has been used to perform pharmacokinetic and bioequivalence studies in humans.
