高效液相色谱-串联质谱法测定比格犬血浆中的莫诺苷浓度及其药代动力学  

Determination of morroniside concentration in beagle plasma and its pharmacokinetics byhighperformance liquid chromatography-tandem mass spectrometry

作　　者：熊山[1,2] 李敬来[2] 朱秀清[2] 王晓英[2] 吕圭源[1] 张振清[2] 

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《色谱》2014年第3期290-293,共4页Chinese Journal of Chromatography

基　　金：科技部"重大新药创制"科技重大专项(2012ZX09301003-001-007)

摘　　要：运用高效液相色谱-串联质谱联用技术，建立了简单、快速、灵敏的比格犬灌胃莫诺苷后血药浓度的检测方法。血浆样品采用蛋白质沉淀法处理，以芍药苷作为内标，色谱柱为 Inertsil ODS-SP 色谱柱（50 mm ×2.1 mm，5μm），流动相为水（含1 mmol / L 甲酸钠）-乙腈，梯度洗脱，流速0.4 mL / min。采用电喷雾离子源（ ESI），正离子多反应监测（MRM）模式。绘制血药浓度-时间曲线，并采用 DAS 2.0软件计算药代动力学参数。方法学实验结果表明内源性杂质不干扰莫诺苷和内标的测定，线性范围为2～5000μg / L（ r =0.9966），定量限为2μg / L。方法精密度、准确度、回收率和基质效应均符合生物样品测定的要求，适合比格犬血浆中莫诺苷浓度的测定，可以应用该方法进行莫诺苷的药代动力学研究。比格犬灌胃莫诺苷3个剂量（5、15、45 mg / kg）后的血药浓度-时间曲线下面积（AUC（0-∞））分别为（1631.20±238.50）、（3984.05±750.38）、（10397.64±3156.34）μg / L＆#183;h，与给药剂量之间呈现良好的线性关系。A sensitive,simple and specific high performance liquid chromatography-electros-pray ionization tandem mass spectrometry（ LC-MS / MS）method was developed for the deter-mination of morroniside in the plasma of beagles administered via intragastric（ ig）doses of morroniside. The method employed paeoniflorin as the internal standard and extracted by sim-ple protein precipitation. The separation was achieved using an Inertsil ODS-SP column（50 mm ＆#215;2. 1 mm,5 μm）with mobile phases of 1 mmol / L sodium formate aqueous solution and aceto-nitrile（gradient elution）at a flow rate of 0. 4 mL / min. The detection was accomplished by a mass spectrometer using multiple reaction monitoring（ MRM）in positive mode. Pharmacoki-netic parameters were fitted by software DAS 2. 0. The methodological study showed a good lin-ear relationship of 2-5 000 μg / L（r = 0. 996 6）with a sensitivity of 2 μg / L as the limit of quanti-fication. The precision,accuracy,mean recoveries and the matrix effects were satisfied with the requirements of biological sample measurement. The method described above was success-fully applied to the pharmacokinetic study of morroniside in the beagle plasma samples. The area under the plasma concentration-time curves（ AUC（ 0-∞ ）） of morroniside after single ig administration doses of 5,15 and 45 mg / kg were（1 631. 20＆#177;238. 50）,（3 984. 05＆#177;750. 38）and （10 397. 64＆#177;3 156. 34）μg / L＆#183;h. The relationship between dose and AUC showed a good linearity. The pharmacokinetic property of morroniside was proposed to be linear pharmacokinetics.

关 键 词：液相色谱-串联质谱 莫诺苷 血浆 比格犬 药代动力学 liquid chromatography-tandem mass SPECTROMETRY ( LC-MS MS ) 

分 类 号：O658[理学—分析化学]