UPLC-MS/MS法测定人血浆中20(S)-原人参二醇的浓度  

Determination of 20(S)-protopanaxadiol Concentration in Human Plasma by UPLC-MS/MS

作　　者：刘胜兰 唐智 陈磊 吴素芬 周玲 廖音娟 张杰[4] LIU Shenglan;TANG Zhi;CHEN Lei;WU Sufen;ZHOU Ling;LIAO Yinjuan;ZHANG Jie(Dept.of Pharmacy,Changsha Hospital of Hunan Normal University/the Fourth Hospital of Changsha,Changsha 410006,China;Hunan Key Laboratory for Bioanalysis of Complex Matrix Samples,Changsha Duxact Biotech Co.,Ltd.,Changsha 410205,China;Central Research Institute,Zhejiang Hisun Pharmaceutical Co.,Ltd.,Zhejiang Taizhou 318000,China;Dept.of Pharmacy,the Third Xiangya Hospital of Central South University,Changsha 410013,China)

机构地区：[1]湖南师范大学附属长沙医院/长沙市第四医院药学部,长沙410006 [2]长沙都正生物科技有限责任公司复杂基质样本生物分析湖南省重点实验室,长沙410205 [3]浙江海正药业股份有限公司中央研究院,浙江台州318000 [4]中南大学湘雅三医院药学部,长沙410013

出　　处：《中国药房》2021年第18期2248-2253,共6页China Pharmacy

基　　金：湖南省科技创新计划项目(No.2017TP1037);湖南创新型省份建设专项(No.2019SK2241);长沙市科技计划项目(No.kq1907011)。

摘　　要：目的:建立测定人血浆中20(S)-原人参二醇(简称为"PPD")浓度的方法。方法:血浆样品经乙腈沉淀蛋白后,以非那雄胺为内标,采用超高效液相色谱-串联质谱法测定。色谱柱为Waters ACQUITY UPLC HSS T3,流动相为5 mmol/L碳酸氢铵溶液-乙腈(梯度洗脱),流速为0.4 mL/min,柱温为40℃,进样量为10μL;离子源为电喷雾离子源,以多反应监测模式(MRM)进行负离子扫描,用于定量分析的离子对分别为m/z 459.40→375.20(PPD)、m/z 371.30→315.30(内标)。同时将该法用于临床样品的测定。结果:PPD检测质量浓度的线性范围为0.25~30.00 ng/mL(r=0.9992),定量下限为0.25 ng/mL;批内、批间RSD均小于10%,相对误差(RE)为-14.61%~12.69%;提取方法和基质效应均不影响PPD的定量测定。人参皂苷CK片100 mg组、人参皂苷CK片200 mg组、人参皂苷CK片300 mg组类风湿性关节炎患者口服相应药物第84天时的平均cmax分别为18.06、30.03、27.00 ng/mL,中位tmax分别为12.0、6.0、12.0 h,平均AUC0-t分别为622.52、668.15、1155.97 ng·h/mL。结论:本研究成功建立了测定人血浆中PPD浓度的方法,且方法灵敏、准确、稳定,操作简便,血浆用量少,可用于临床样本的定量测定。OBJECTIVE:To establish the method for the determination of 20(S)-protopanaxadiol(PPD)concentration in human plasma.METHODS:Plasma samples were precipitated with acetonitrile and determined by UPLC-MS/MS,using finandrogen as internal standard.The determination was performed on Waters ACQUITY UPLC HSS T3 column with mobile phase consisted of 5 mmol/L ammonium bicarbonate aqueous solution-acetonitrile(gradient elution)at the flow rate of 0.4 mL/min.The column temperature was set at 40℃,and sample size was 10μL.The ion source was electrospray ion source,and negative ion scanning was carried out with multiple reaction monitoring mode.The ion pairs used for quantitative analysis were m/z 459.40→375.20(PPD)and m/z 371.30→315.30(internal standard).At the same time,the method was applied to the determination of clinical samples.RESULTS:The linear range of PPD was 0.25-30.00 ng/mL(r=0.9992),and the limit of quantitation was 0.25ng/mL.RSDs of intra-batch and inter-batch were all lower than 10%,and relative errors(RE)were-14.61%-12.69%.Extraction method and matrix effect did not affect the quantitative determination of PPD.In ginsenoside CK 100 mg group,ginsenoside CK200 mg group and ginsenoside CK 300 mg group,mean cmaxof patients with rheumatoid arthritis after oral administration of corresponding drugs were 18.06,30.03,27.00 ng/mL;median tmaxwere 12.0,6.0,12.0 h;mean AUC0-t were 622.52,668.15,1155.97 ng·h/mL.CONCLUTIONS:The method for the determination of PPD concentration in human plasma is successfully established.The method is sensitive,accurate,stable,easy to operate and less plasma consumption.It can be used for the quantitative determination of clinical samples.

关 键 词：20(S)-原人参二醇 超高效液相色谱-串联质谱法 蛋白沉淀法 血药浓度 

分 类 号：R917[医药卫生—药物分析学] R593.22[医药卫生—药学]