Liquid Chromatography/Tandem Mass Spectrometry Method for Estimation of Cholic Acid in Rat Plasma, Urine and its ApplicationSurendra Yadav R1, 2* and Kilari Eswar Kumar3
- *Corresponding Author:
- Surendra Yadav R
DMPK Laboratory (Biology Division), GVK BIO, Nacharam
Hyderabad, Andhra Pradesh, India-500076
E-mail: [email protected]
Received date: July 13, 2014; Accepted date: August 11, 2014; Published date: August 14, 2014
Citation: Yadav SR, Kumar KE (2014) Liquid Chromatography/Tandem Mass Spectrometry Method for Estimation of Cholic Acid in Rat Plasma, Urine and its Application. J Anal Bioanal Tech 5: 200. doi: 10.4172/2155-9872.1000200
Copyright: © 2014 Yadav SR, 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.
Cholic acid is a primary bile acid synthesised from cholesterol in liver. Apart from being major catabolic products of cholesterol bile acids facilitates absorption of fats in intestine. A selective, sensitive MRM based liquid chromatography/ tandem mass spectrometry method has been developed and validated for the estimation of cholic acid in rat plasma and urine. Telmisartan was used as internal standard (IS). Electrospray ionization (ESI) probe with negative mode of operation was used for the ionisation of cholic acid and IS. Chromatographic separation was performed through X-bridge C18 column with the mobile phase of 0.1% formic acid in water (aqueous reservoir), 100% methanol (organic modifier). A short LC gradient of 3 minutes run time was used with flow rate of 0.7 mL/min. Charcoal stripped plasma and urine were used for the preparation of calibration standards and quality control samples. The analyte and IS were isolated from plasma and urine by a simple organic solvent based protein precipitation. The assay was linear in the concentration range of 31.26-10000 ng/mL. The method has been validated according to published FDA guidelines and showed excellent performance. The developed method was applied for the estimation of cholic acid in plasma and urine samples from liver toxicity experiment in rats. Liver toxicity was induced by intraperitoneal injection of carbon tetrachloride (CCl4).