Page 78 - 2021年18期

·药物与临床·

UPLC-MS/MS法测定人血浆中20（S）-原人参二醇的浓度 Δ

刘胜兰 ，唐 智 ，陈 磊 ，吴素芬 ，周 玲 ，廖音娟 ，张 杰（1.湖南师范大学附属长沙医院/长沙市第四医
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1＊
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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）
中图分类号 R917；R593.22 文献标志码 A 文章编号 1001-0408（2021）18-2248-06
DOI 10.6039/j.issn.1001-0408.2021.18.12

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10 μL；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＝0.999 2），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、1 155.97 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。
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

Determination of 20（S）-protopanaxadiol Concentration in Human Plasma by UPLC-MS/MS
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LIU Shenglan ，TANG Zhi ，CHEN Lei ，WU Sufen ，ZHOU Ling ，LIAO Yinjuan ，ZHANG Jie（1. Dept. of
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Pharmacy， Changsha Hospital of Hunan Normal University/the Fourth Hospital of Changsha， Changsha
410006，China；2. Hunan Key Laboratory for Bioanalysis of Complex Matrix Samples，Changsha Duxact
Biotech Co.，Ltd.，Changsha 410205，China； 3. Central Research Institute，Zhejiang Hisun Pharmaceutical
Co.，Ltd.，Zhejiang Taizhou 318000，China；4. Dept. of Pharmacy，the Third Xiangya Hospital of Central
South University，Changsha 410013，China）
ABSTRACT 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 10 μL. 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＝0.999 2），and the limit of quantitation was 0.25
ng/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 CK
200 mg group and ginsenoside CK 300 mg group，mean cmax of patients with rheumatoid arthritis after oral administration of
corresponding drugs were 18.06，30.03，27.00 ng/mL；median tmax were 12.0，6.0，12.0 h；mean AUC0-t were 622.52，668.15，
1 155.97 ng·h/mL. CONCLUTIONS：The method for the
Δ 基金项目：湖南省科技创新计划项目（No.2017TP1037）；湖南创
determination of PPD concentration in human plasma is
新型省份建设专项（No.2019SK2241）；长沙市科技计划项目（No.
successfully established. The method is sensitive，accurate，
kq1907011）
stable，easy to operate and less plasma consumption. It can be
＊药师，硕士。研究方向：体内药物分析。E-mail：384325419@
used for the quantitative determination of clinical samples.
qq.com
KEYWORDS 20（S）-protopanaxadiol；UPLC-MS/MS；Protein
# 通信作者：副主任药师，硕士。研究方向：临床药学、医院药事
管理。电话：0731-88942106。E-mail：348785608@qq.com precipitation method；Plasma concentration

·2248 · China Pharmacy 2021 Vol. 32 No. 18 中国药房 2021年第32卷第18期