Toxicol Res.  2013 Sep;29(3):203-209.

Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of epsilon-Acetamidocaproic Acid in Rat Plasma

• Kim TH
• Choi YS
• Choi YH
• Kim YG

Abstract

A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of epsilon-acetamidocaproic acid (AACA), the primary metabolite of zinc acexamate (ZAC), in rat plasma by using normetanephrine as an internal standard. Sample preparation involved protein precipitation using methanol. Separation was achieved on a Gemini-NX C18 column (150 mm x 2.0 mm, i.d., 3 microm particle size) using a mixture of 0.1% formic acid-water : acetonitrile (80 : 20, v/v) as the mobile phase at a flow rate of 200 microl/min. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 4.0 min, and the calibration curves of AACA were linear over the concentration range of 20~5000 ng/ml in rat plasma. The coefficient of variation and relative error at four QC levels were ranged from 1.0% to 5.8% and from -8.4% to 6.6%, respectively. The present method was successfully applied for estimating the pharmacokinetic parameters of AACA following intravenous or oral administration of ZAC to rats.

Keyword

Zinc acexamate (ZAC); epsilon-Acetamidocaproic acid (AACA); Liquid chromatography tandem mass spectrometry (LC-MS/MS); Rat; Pharmacokinetics

MeSH Terms

6-Aminocaproic Acid
Acetonitriles
Administration, Oral
Animals
Calibration
Mass Spectrometry*
Methanol
Normetanephrine
Pharmacokinetics
Plasma*
Rats*
Zinc
6-Aminocaproic Acid
Acetonitriles
Methanol
Normetanephrine
Zinc