Yahdiana Harahap
Universities Indonesia, Indonesia
Title: The effect of anticoagulant types in analyzing levofloxacin in human plasma by high performance liquid chromatography-photodiode array
Biography
Biography: Yahdiana Harahap
Abstract
Levofloxacin has low concentration in plasma, thus it requires sensitive and selective analysis method. Plasma drug analysis often uses many kinds of anticoagulant to obtain plasma as analytical matrix. Citrate, heparin, and ethylenediaminetetraacetic acid (EDTA) are anticoagulant commonly used in analyzing drug in human plasma. This study was focused on analyzing levofloxacin in human plasma with three types of anticoagulants. The analysis was performed using High Performance Liquid Chromatography (HPLC) – photodiode array with Column C18 SunfireTM (250 x 4.6mm), 5ïm; temperature of 45ï¯C, mobile phase consist of 0.5% triethylamine pH 3.0 -acetonitrile (88:12 v/v); flow rate of 1.25 mL/minute, and ciprofloxacin HCl as internal standard. The method was linear at concentration range of 50.0 – 10.000.0 ng/mL with r>0.9994. Accuracy and precision for citrate, heparin, and EDTA plasma fulfilled the acceptance criteria of both intra-day and inter-day. There was no significant difference for stability and recovery of levofloxacin in citrate, heparin, and EDTA plasma (p>0.05; ANOVA), but it showed significant difference for peak area ratio (p>0.05), between citrate-EDTA plasma and heparin-EDTA plasma for low concentration and between citrate-heparin plasma and citrate-EDTA plasma for mid and high concentration. On blank chromatogram EDTA plasma, there was interference on retention time of less than 8 minutes, while on citrate and heparin plasma there was no interference. The method can be applied for bioequivalence study using the three anticoagulants that are equally good.