UPLC-MS/MS法测定犬血浆中的雷公藤次碱及体内药动学研究  

Determination of Wilforine in the Beagle dogs Plasma by UPLC-MS/MS and on its Pharmacokinetics

作　　者：杨炀 宋小仙[1] 涂如霞[1] 兰波[1] 吴思澜 莫宗成[1] 罗金萍[1] 张莉[1] 刘剑毅[1] Yang Yang;Song Xiaoxian;Tu Ruxia;Lan Bo;Wu Silan;Mo Zongcheng;Luo Jinping;Zhang Li;Liu Jiangyi(Chongqing Academy of Chinese Materia Medica,Chongqing,400065 P.R.China)

机构地区：[1]重庆市中药研究院,重庆400065

出　　处：《世界科学技术-中医药现代化》2020年第6期1940-1946,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：重庆市科学技术局重庆市科研院所绩效激励引导专项(cstc2018jxjl130055):雷公藤次碱在Beagle犬体内的代谢动力学研究,负责人:莫宗成。

摘　　要：目的建立超高效液相-串联质谱(UPLC-MS/MS)法测定犬血浆中的雷公藤次碱的浓度,并用于雷公藤次碱的体内药代动力学研究。方法色谱柱为ACE Super C18柱(2.1×100 mm id,5μm,菲罗门),以2 mM醋酸铵(0.2%甲酸)-乙腈为流动相,梯度洗脱,流速为350μL·mL^-1,柱温30℃,进样量2μL。以雷公藤春碱为内标,血浆样本前处理后,采用ESI-Positive-MRM模式,进样检测。将Beagle犬分为高中低剂量组(400、200、100μg·kg^-1),给药后测定不同时间点犬血浆中雷公藤次碱的浓度,采用DAS3.0软件对雷公藤次碱药时曲线进行拟合,并计算药代动力学参数。结果建立的UHPLC-MS/MS法在5.0-250 ng·mL^-1范围内雷公藤次碱色谱响应与浓度相关性良好,定量限为4.94 ng·mL^-1,雷公藤次碱及内标的保留时间分别为2.99、2.73 min,该方法无明显的离子抑制或增强效应,批内批间精密度及准确度RSD均小于15%,提取回收率在82.86%^-112.76%之间,稳定性各项数据显示中、高浓度RSD均小于15%,低浓度RSD均小于20%,均符合要求。雷公藤次碱吸收较快,在Beagle犬体内的系统暴露随剂量的增大而增加,有呈比例增加的趋势;其消除与性别有关,与剂量无关。结论该检测方法灵敏度高、专属性强、准确度良好,适用于犬血浆中雷公藤次碱的浓度测定及其药动学研究。Objective To establish an UPLC-MS/MS method for the determination of wilforine in the Beagle dogs plasma,and to study its pharmacokinetics.Methods The separation was performed on a reverse phase Phenomenex ACE Super C18 columu(2.1×100 mm id,5μm),with the mobile phase gradient elution consisted of elution of acetonitrile-water(contain 0.2%formic acid and 2 mM Ammonium acetate)at a flow rate of 350μL·min^-1.Wilfortrine was used as internal standard.After pretreatment of plasma samples,the ESI-Positive-MRM mode was used for injection detection.The Beagle dogs were divided into high、medium and low-dose groups(400,200,100μg·Kg^-1).The concentration of wilforine in canine plasma was measured at different time points after administration.The DAS3.0 software was used for wilforine.The curves were fitted and the pharmacokinetic parameters were calculated.Results The UPLC-MS/MS method has a good correlation between the chromatographic response and concentration of wilforine in the range of 5 to 250 ng·mL^-1.The retention times of wilforine and internal standard were 2.99 and 2.73 min,respectively.The method has no obvious ion suppression or enhancement effect,and the precision and accuracy are good,and the stability data meets the requirements.The absorption of tripterine was rapid,and the systemic exposure in Beagle dogs increased with the increase of dose,which showed a trend of proportional increase.Its elimination was related to gender and independent of dose.Conclusion The mehod is accurate,specificitive and sensitive,and it is suitable for the concentration determining of wilforine in the Beagle dogs plasma and its pharmacokinetic study.

关 键 词：雷公藤次碱 UPLC-MS/MS 药动学 血药浓度 含量测定 

分 类 号：R285[医药卫生—中药学]