Last modified: 2019-06-17
Abstract
Effect of Various Anticoagulants on Amlodipine Besylate and Valsartan for Simultaneous Analysis in Human Plasma by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry
Maria Romauli, Yahdiana Harahap, Sunarsih
Faaculty of Pharmacy, Universitas Indonesia, Depok 16424, West Java Indonesia
Corresponding author :
yahdiana03@yahoo.com
Amlodipine besylate and valsartan two compounds that are commonly found in a drug tablet for anti-hypertensive therapy available on pharmacy counters. Furthermore, this medication is known to have low concentration in plasma, therefore an selective and sensitive method to determine it. In order to obtain plasma from blood, an addition of anticoagulant is needed. A research about in vitro study for method development and validation method analysis used citrate as anticoagulant. However, anticoagulants commonly used for in vivo study are EDTA and heparin. The anticoagulant difference allows it to interfere with the analysis so that partial validation is needed. This research objective was to evaluate the different types of anticoagulant for amlodipine besylate and valsartan in plasma using ultra performance liquid chromatography tandem mass spectrometry. The chromatography condition used BEH C18 Waters column (2.1 × 100 mm; 1.7 μm); mobile phase consisting of 0.1% formic acid in water – acetonitrile with gradient elution; flow rate of 0.2 mL/minute; injection volume of 10 μL; column temperature of 45oC; 6 minutes analysis time and irbesartan as internal standard. The sample preparation used liquid-liquid extraction with ammonium acetate pH 4.83 and ethyl acetate as solvent. The accuracy and precision of the analysis of citrate, heparin and EDTA plasma met the requirements and linear calibration curve at concentrations range of 0.2-10 ng / mL for amlodipine besylate and 5-6000 ng/mL for valsartan. The stability and peak area ratio of each plasma was evaluated using ANOVA.
Keywords : citrate, EDTA, heparin, ultra performance liquid chromatography, amlodipine besylate, valsartan