反相离子对高效液相色谱法测定血浆吗啡及其代谢物M3G的含量

目的:建立反相离子对高效液相色谱法测定血浆吗啡及其代谢物3-葡萄糖醛酸吗啡(M3G)的浓度。方法:血浆样品以乙酸乙酯提取,以纳洛酮为内标。应用HPLC-UV方法测定吗啡浓度,分析柱DiamonsilTMC18(250mm×4.6mm),流动相为0.01mol·L-1磷酸二氢钾缓冲液(含0.5mmol·L-1的十二烷基磺酸钠)-乙腈(71:29),检测波长210nm,流速0.65ml·min1。血浆样品经β-葡萄糖醛酸苷酶水解测定其代谢物含量。结果:吗啡标准曲线线性范围宽(10-4000μg·L-1),线性关系良好;最低检测限10μg·L-1;高、中、低浓度的回收率在85·3%以上。血浆M3G在250-8250μg·L-1浓度范围内与水解增加吗啡浓度呈线性关系。结论:本法简单,快速,血浆中杂质不干扰样品的测定,满足生物样品分析要求。

DETERMINATION OF MORPHINE AND METABOLITE M3G IN RAT PLASMA BY REVERSED-PHASE IRON-PAIR HPLC

Objective:To establish a reversed-phase iron-pair HPLC for determination of morphine and metabolite morphine-3-glucuronide(M3G) in rat plasma. Methods:Morphine was extracted from rat plasma using ethyl acetate,then determinated by HPLC-UV afterseparationon a Diamonsil~ TM C_ 18 analytical column (250mm×4.6 mm). The mobile phase was 0.01 mol·L~ - 1 potassium dihydrogen phosphate (0.5 mmol[DK]·[DK]L~ - 1 sodium dodecylsulfonate)–acetonitrile (71[DK]∶[DK]29),the wavelength of detection was set at 210 nm, and flow rate was 0.65 ml[DK]·[DK]min~ -1 . Internal standard was naloxone. Plasma was hydrolyzed by β-glucuronidase for determination metabolite M3G. Results:Linearity was verified from 10 to 4000 μg[DK]·[DK]L~ -1 , the limit of quantition(LOQ) was 10 μg[DK]·[DK]L~ -1 .The recovery of morphine was above 85.3%. Linearity was also verified between the concentration of M3G (from 250 to 8250 μg[DK]·[DK]L~ -1 )and morphine obtained from M3G in the plasma. Conclusion:This method is simple, rapid and accurate for quantitative analysis of morphine in rat plasma.