Open AccessEFFECT OF DIFFERENT ANTICOAGULANTS IN DETERMINING METFORMIN HYDROCHLORIDE LEVELS IN HUMAN PLASMA USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Author(s) -
Yahdiana Harahap,
Lista Roro Marsudi,
Sunarsih Sunarsih
Publication year - 2018
Publication title -
international journal of applied pharmaceutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.238
H-Index - 15
ISSN - 0975-7058
DOI - 10.22159/ijap.2018.v10s1.31
Subject(s) - chromatography , chemistry , ethylenediaminetetraacetic acid , protein precipitation , high performance liquid chromatography , metformin hydrochloride , acetonitrile , calibration curve , bioanalysis , metformin , detection limit , chelation , medicine , organic chemistry , diabetes mellitus , endocrinology
Objective: This study aimed to develop and validate an analytical method and to determine the effect of different anticoagulants for analyzingmetformin HCl level in human plasma.Methods: Analysis was performed with high-performance liquid chromatography method using a photodiode array detector set at 234 nm, a C18SunFire™ column (5 μm, 250 mm×4.6 mm), a temperature of 40°C, a mobile phase comprising 10 mM sodium dodecyl sulfate and 10 mM phosphatebuffer in water-acetonitrile (60:40 v/v) at pH 7.0, with a flow rate of 1.0 mL/min, and atorvastatin calcium as the internal standard. Plasmaextraction was performed through protein precipitation using human plasma (300 μL) and acetonitrile (600 μL; 1:2 v/v). The method was linear at aconcentration range of 20.0–5000.0 ng/mL, with r>0.9998.Results: The stability and recovery of metformin HCl in human plasma were not different between citrate with ethylenediaminetetraacetic acid(EDTA) and heparin with EDTA as anticoagulants. However, a significant difference in the peak area response ratio (p<0.05) was observed betweenthe three anticoagulants.Conclusion: The validation results for the three types of plasma anticoagulants met validation requirements based on the European MedicinesAgency bioanalytical guidelines of 2011 for accuracy and precision, selectivity, calibration curve linearity, dilution integrity, carry-over, and stability.