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Abstract：
Semen Strychni is known for its treatment of rheumatic arthritis with a low therapeutic index. Liquorice contributes a lot in herb detoxification according to the traditional Chinese medicine theory. A simple, rapid, and sensitive liquid chromatography-mass spectrometric method (LC-MS) was developed and validated for simultaneous determination of main bioactive ingredients in liquorice and Semen Strychni in rat plasma. Using moclobemide and cyproterone acetate as the internal standards, the analytes were pretreated via protein precipitation with methanol. An Ultimate AQ-C18 column (3.0 μm, 3.0 × 100 mm) was employed for chromatographic separation, combining with gradient elution. The mobile phase consisted of 0.07% formic acid and 0.12% ammonium acetate in aqueous phase (A) and acetonitrile in organic phase (B). The elution program was as follows: 0-0.5 min, 20% B; 0.5-1 min, 20-60% B; 1-7 min, 60-85% B; and 7-7.5 min, returned to 20% B, then continued to 12 min. Selected reaction monitoring was performed in both positive and negative ESI. Positive mode was adopted for detection of strychnine, brucine, and moclobemide, while negative mode was used for glycyrrhizic acid, glycyrrhetinic acid, liquiritigenin, isoliquiritigenin, liquiritin, and cyproterone acetate. The method was validated for specificity, linearity, matrix effect, recovery, precision, accuracy, and stability. The results show that this method is sensitive, accurate and robust for biological matrix analysis. Moreover, the proposed method was applied to a pharmacokinetic study in Sprague-Dawley rats for investigating the mechanism of which liquorice detoxifies Semen Strychni.
摘要：
精液Strychni以其低治疗指数治疗风湿性关节炎而闻名。根据中医理论，甘草在中药解毒中的作用很大。一个简单的，快速,建立了灵敏的液相色谱-质谱（LC-MS）方法，并验证了该方法可同时测定大鼠血浆中甘草和马钱子的主要生物活性成分。使用莫洛贝胺和醋酸环丙孕酮作为内标，通过用甲醇进行蛋白质沉淀对分析物进行预处理。最终AQ-C18色谱柱(3.0μm,采用3.0×100mm)进行色谱分离，结合梯度洗脱。流动相由在水相(A)中的0.07%甲酸和0.12%乙酸铵和在有机相(B)中的乙腈组成。洗脱程序如下：0-0.5分钟，20%B;0.5-1分钟,20-60%B;1-7分钟,60-85%B；和7-7.5分钟，恢复到20%B，然后继续12分钟。在阳性和阴性ESI中均进行选择的反应监测。采用阳性模式检测士的宁，马钱子碱,和莫洛贝胺,而甘草酸使用阴性模式，甘草次酸，甘草素,异甘草素,甘草苷,和醋酸环丙孕酮.对该方法的特异性进行了验证，线性度基体效应，recovery,精度,准确度,和稳定性。结果表明,该方法灵敏度高,生物基质分析的准确性和鲁棒性。此外，将该方法应用于Sprague-Dawley大鼠的药代动力学研究，以研究甘草排毒使精液Strychni的机制。
