超高效液相色谱串联质谱法测定大鼠血浆中薯蓣皂苷元浓度  

Determination of Diosgenin in Rat Plasma by UPLC-MS/MS

作　　者：常金花[1] 刘喜纲[1] 薛禾菲 张琳[1] 刘沛[1] 刘冰洋[2] 付强[2] 刘翠哲[1] 

机构地区：[1]河北省中药研究与开发重点实验室,承德医学院,河北承德067000 [2]沈阳药科大学,沈阳110016

出　　处：《中国药学杂志》2017年第17期1536-1541,共6页Chinese Pharmaceutical Journal

基　　金：河北省自然科学基金资助项目(H2014406036);河北省高等学校科学技术研究资助项目(QN2015127);河北省高校重点学科建设资助项目(冀教高[2013]4号)

摘　　要：目的建立超高效液相色谱串联质谱法测定大鼠血浆中薯蓣皂苷元浓度。方法以丹参酮ⅡA为内标。血浆经甲醇沉淀蛋白后,采用色谱柱为Phenomenex kinetex xb C_(18)(2.1 mm×50 mm,2.6μm),以甲醇(含0.1%甲酸)(A)-0.1%甲酸(B)为流动相,梯度洗脱,流速为0.2 m L·min^(-1),柱温为40℃,进样量为5μL;以电喷雾(ESI)离子源,在正离子电离模式下,采用多反应监测(MRM)的扫描模式进行测定。薯蓣皂苷元和丹参酮ⅡA的MRM扫描离子对分别为m/z 415.2→271.1和m/z295.1→249.1。结果血浆中薯蓣皂苷元质量浓度在10~500 ng·mL^(-1)内线性关系良好(r=0.998 3);最低定量限为10 ng·mL^(-1);准确度为96.1%~102.3%;日内和日间精密度均小于15%;平均提取回收率为73.8%~75.2%;基质效应为85.8%~91.7%。内标的平均提取回收率为83.8%,基质效应为92.4%。大鼠灌胃给予薯蓣皂苷元(100 mg·kg^(-1))后,薯蓣皂苷元在大鼠体内的达峰浓度为(344.067±34.48)ng·mL^(-1),达峰时间为(4.167±2.041)h,半衰期为(14.85±10.53)h,血药浓度-时间曲线下面积为(4 965.648±1 036.129)μg·h·L^(-1)。结论该方法专属性强,样品处理方便,灵敏度高,检测时间短,适用于薯蓣皂苷元在大鼠体内的药动学研究。OBJECTIVE To develop and validate a sensitive and specific ultra-performance liquid chromatography-tandemmass spectrometric （LC-MS/MS） method for the assay of diosgenin in rat plasma. METHODS Tanshinone II A was employed as internal standard. Diosgenin was determined after the methanol-mediated plasma protein precipitation. The separation was performed on the Phenomenex kinetex xb C,s column （2. 1 mm x 50 mm ,2. 6 μm） gradiently eluted with the mobile phase consisting of methanol（ contai- ning 0. 1% formic acid） -0. 1% aqueous formic acid. The flow rate was 0. 2 mL · min-1, the column temperature was maintained at 40 ℃, and the injection volume was 5 μL A triple quadrupole mass spectrometer equipped with electrospray ionization source was used as detector in a positive ion mode. Multiple reaction monitoring （MRM） mode was applied with the transition of m/z 415.2-271.1 and m/z 295.1-249. 1 for diosgenin and internal standard, respectively. RESULTS For diosgenin the standard curve was linear from 10 to 500 ng· mL-1 （r =0. 998 3） , the limit of quantitative limit was 10 ng· mL-1 , the intra- and inter-assay variabilities were below 15% , the accuracies were between 96. 1% and 102. 3%, the average extract recoveries ranged from 73.8% to 75.2%, and the matrix effects was between 85.8% and 91.7%. For the internal standard, the extract recovery and matrix effects were 83.8% and 92. 4% , respectively. The rats were administered orally with diosgenin （ 100 mg · kg-l ） . The peak concentration of diosgenin was （344. 067 ± 34.48 ） ng · mL- 1, the time for peak concentration was （4. 167 ± 2. 041 ） h, the half-time was （ 14. 85 ± 10. 53 ） h, and the area under concentration-time curve from zero to 72 h was （4 965. 648 ± 1 036. 129） μg · h · L-1. CONCLUSION This assay is specific, simple, sensitive and rapid, which can be applied in the pharmacokinetic study of diosgenin in rats.

关 键 词：薯蓣皂苷元 血药浓度 药动学 超高效液相色谱串联质谱法 

分 类 号：R917[医药卫生—药物分析学]