液相色谱串联质谱法测定小鼠血浆中人参皂苷Rh2浓度  

Determination of ginsenoside Rh2 in plasma of mice by liquid chromatography-tandem mass spectrometry

作　　者：席鑫 丁新月 法菁菁 黄新苗[2] 刘宗军[1] XI Xin;DING Xinyue;FA Jingjing;HUANG Xinmiao;LIU Zongjun(Department of Cardiovasology,Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Department of Cardiovasology,The First Affiliated Hospital of Naval Medical University(Second Military Medical University),Shanghai 200433,China)

机构地区：[1]上海中医药大学附属普陀医院心血管内科,上海200062 [2]海军军医大学(第二军医大学)第一附属医院心血管内科,上海200433

出　　处：《海军军医大学学报》2025年第2期268-272,共5页Academic Journal of Naval Medical University

基　　金：上海市科学技术委员会科技创新行动计划(21Y11909600).

摘　　要：目的建立液相色谱串联质谱法(LC-MS/MS)测定小鼠血浆中人参皂苷Rh2(GRh2)血药浓度,为GRh2的药代动力学研究及应用提供临床前数据支持。方法C57BL/6小鼠灌胃7.5 mg/kg GRh2,于给药后5 min、10 min、15 min、30 min、1 h、2 h、4 h、8 h收集全血0.03 mL,全血离心分离血清后用0.1%甲酸乙腈处理,经氮气(40℃)吹干后用50.0µL含100 ng/mL双氯芬酸钠的50%甲醇溶液复溶,室温条件下涡旋混匀5 min后放入自动进样器中进样分析。色谱柱为Waters BEHC18(2.1 mm×50.0 mm,1.7μm),流动相为0.1%甲酸水溶液和含0.1%甲酸的乙腈溶液,流速为0.60 mL/min,柱温40℃,进样体积1.00µL。采用电喷雾离子源,正离子模式,多反应监测。建立标准曲线,计算血药浓度并建立血药浓度-时间曲线,计算主要药代动力学参数。结果含药血浆标准曲线线性范围为100~40000 ng/mL,相关系数(r)为0.9960。内标归一化后GRh2基质效应因子分别为1.09、1.06、1.00(在0.8~1.2之间),表明无明显的基质效应。精密度和准确度结果显示GRh2每一浓度水平样品的平均实测浓度为103、333、23800、35000 ng/mL,平均批间标准差在6.47~1120 ng/mL,批间RSD在1.5%~8.3%,平均批间准确度偏差在93.3%~111.1%。GRh2的长期稳定性、短期稳定性、反复冻融性、提取回收率均良好。药代动力学实验结果显示GRh2灌胃给药小鼠体内的药代动力学参数Tmax、Cmax分别为(1.42±1.01)h、(1251±495)ng/mL,表明GRh2的体内吸收利用率较高,具有良好的成药性。结论所建立的LC-MS/MS准确、可靠,可用于小鼠血浆中GRh2的浓度测定及其药代动力学研究。Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of ginsenoside Rh2(GRh2)in plasma of mice,so as to provide preclinical data support for the pharmacokinetic study and application of GRh2.Methods C57BL16 mice were given 7.5 mg/kg GRh2 by gavage.After administration,0.03 mL of whole blood was collected at 5 min,10 min,15 min,30 min,1 h,2 h,4 h,and 8 h.Then,the whole blood was centrifuged and the serum was treated with 0.1%formic acid acetonitrile,dried by nitrogen(40℃),and redissolved with 50.0µL 50%methanol solution containing 100 ng/mL diclofenac sodium.After vortex mixing for 5 min at room temperature,it was put into the automatic sampler for sampling analysis.The chromatographic column was Waters BEHC18(2.1 mm×50.0 mm,1.7μm),the mobile phase was aqueous solution containing 0.1%formic acid and acetonitrile solution containing 0.1%formic acid at a flow rate of 0.60 mL/min,the column temperature was 40℃,and the injection volume was 1.00µL.The electric spray ion source,positive ion mode and multiple reaction monitoring mode were performed.A standard curve was established to calculate blood drug concentration.The blood drug concentration-time curve was established according to the blood drug concentration,and the main pharmacokinetic parameters were calculated.Results The linear range of the standard curve of drug containing plasma was 100-40000 ng/mL,and the correlation coefficient(r)was 0.9960.After internal standard normalization,the matrix effect factors of GRh2 were 1.09,1.06,and 1.00(between 0.8 and 1.2),indicating no significant matrix effect.The precision and accuracy results showed that the average measured concentration of GRh2 samples at each concentration level was 103,333,23800 and 35000 ng/mL,the inter batch standard deviation was 6.47-1120 ng/mL,the inter batch relative standard deviation was 1.5%-8.3%,and the inter batch accuracy deviation was 93.3%-111.1%.The longterm stability,short-term stability,repeated freeze-thaw property,and

关 键 词：人参皂苷RH2 液相色谱串联质谱法 药代动力学 血药浓度 

分 类 号：O657.63[理学—分析化学] R285.5[理学—化学]