反相高效液相色谱法测定人血浆洛拉曲克浓度

Concentration Determination of Nolatrexed in Human Plasma by RP-HPLC

目的建立测定人血浆洛拉曲克浓度的反相高效液相色谱法。方法以DiamonsilTM C18反相柱（150 mm×4.6 mm, 5 μm）为色谱柱,流动相为0.03 mol·L-1醋酸铵-甲醇（40：60）;流速：0.8 mL·min-1;柱温：40 ℃;检测波长： 233 nm。以乙酸乙酯与二氯甲烷（80：20）为提取剂。结果洛拉曲克的高、中、低（50.0, 10.0,1.0 μg·mL-1）3种浓度平均回收率分别为98.72%,97.86%,101.27%,日内、日间差RSD均＜7%（n＝5）;分析方法的检测限为1.0 μg·mL-1;线性范围为1.0～50.0 μg·mL-1。标准曲线方程： Y＝1.73X－5.29,r＝0.999 8（n＝8）。结论该方法灵敏、准确、简单、快速,可用于临床血药浓度监测和药动学研究。

Objective To establish a method for determining the concentration of nolatrexed in human plasma by RP-HPLC. MethodsThe drug from plasma was analyzed in a RP-HPLC system： C18 column（150 mm×4.6 mm, 5 μm）; mobile phase consisted of 0.03 mol·L-1 ammonioum-methanol（40：60）; the flow rate at 0.8 mL·min-1; the detection wavelength at 233 nm. Ethyl acetate and dichloromethane（80：20） were used as extracting solvent. ResultsThe average recoveries of nolatrexed in high, middle and low concentrations （50.0, 10.0, 1.0 μg·mL-1） were 98.72% , 97.86% and 101.27%, respectively. The intra-day and inter-day variation（RSD）was less than 7% （n=5）. The calibration curve of nolatrexed showed good linearity, r=0.999 8（n=8）, over the range of 1.0-50.0 μg·mL-1. The regression equation was Y=1.73X-5.29. r=0.999 8（n=8）. The minimum detectable concentration of nolatrexed was 1.0 μg·mL-1. ConclusionThe method is sensitive, accurate, easy and quick, which can be used for clinical drug monitoring and pharmacokinetics studies.