P-HPLC法测定小鼠血清及脏器中拉米夫定的浓度

Rapid Quantitation of Lamivudine in Mouse Serum and Tissue Homogenates by RP-HPLC

目的 :建立RP -HPLC法快速测定小鼠血清及部分脏器匀浆中拉米夫定 (LAM )的浓度。方法 :取0 1ml血清或组织匀浆 ,加入0 5ml甲醇 ,漩涡混合30s ,15000r/min离心10min ,上清液直接进样。选用KromasilC18(150mm×4 6mm ,5μm)为分析柱 ,pH6 8的磷酸盐缓冲液 -甲醇 (91∶9)为流动相 ,检测波长为270nm。结果 :各工作曲线线性范围为0 25～50μg/ml ,最低检测浓度为60ng/ml(S/N=3)。各样品平均绝对回收率大于88 % ,方法回收率在97 %～100 %之间 ,日内和日间精密度的RSD均小于10 %。结论 :此方法经济、可靠、简单、快速 。

OBJECTIVE:To develop a rapid RP-HPLC quantitative determination of lamivudine(LAM) in mouse serum and tissue homogenates METHODS:0 1ml of serum of tissue homogenates and 0 5ml of methanol were vortexed in a plastic tube for 30s and the mixture was then centrifuged at 15 000r/min for 10min 20μl of the resulting supernatant was directly injected and LAM was chromatographed on a reversed-phase Kromasil C column(150mm×4 6mm,5μm) using a mixture of phosphate buffer(pH6 8)and methanol(91∶9,v/v)and monitored at 270nm RESULTS:The standard curves were linear over a range from 0 25μg/ml to 50μg/ml The minimal detectable drug concentration was 60ng/ml(S/N=3) The average extraction recovery of LAM was more than 88% The average methodological recoveries of LAM ranged from 97% to 100% The within-dayand between-day precisions were less than 10% CONCLUSION:The assay is economic,simple,rapid and specific