Page 53 - 《中国药房》2025年13期

大鼠血浆中氯吡格雷及其代谢物的测定方法建立与药动学研究
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易 欢 ，苗 兰，任常英，林 力，孙明谦，彭 勍，张 颖 ，刘建勋（中国中医科学院西苑医院基础医学研究所/
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北京市中药药理重点实验室，北京 100091）

中图分类号 R969.1 文献标志码 A 文章编号 1001-0408（2025）13-1599-05
DOI 10.6039/j.issn.1001-0408.2025.13.08

摘 要 目的 建立大鼠血浆中氯吡格雷（CLP）、氯吡格雷羧酸（CLP-C）、氯吡格雷酰基-β-D-葡糖醛酸（CLP-G）以及含巯基的氯
吡格雷活性代谢产物（CAM）的含量测定方法，并研究其体内药动学特征。方法 以Shisedo CAPCELL ADME为色谱柱，水和乙腈
（均含0.1%甲酸）为流动相进行梯度洗脱，流速为0.4 mL/min，柱温为20 ℃，进样量为2 μL；质谱检测采用电喷雾离子源，在正离
子模式下进行多反应监测，离子对分别为m/z 322.1→211.9（CLP）、m/z 308.1→197.9（CLP-C）、m/z 322.1→154.8（CLP-G）、m/z 504.1→
154.9[外消旋CAM衍生物（CAMD）]。取6只大鼠单次灌胃CLP（10 mg/kg），分别于给药前和给药后0.08、0.33、0.66、1、2、4、6、10、
23、35 h时收集血样，采用上述建立的方法检测血清中各成分含量，并利用WinNonlin 6.1软件计算药动学参数。结果 CLP、CLP-C、
CAMD 的质量浓度分别在 0.08～20.00、205.00～8 000.00、0.04～25.00 ng/mL 范围内线性关系良好（r≥0.990），日内、日间精密度
试验的RSD均小于15%，准确度的RE为－11.68%～14.40%，基质因子的变异系数均小于15%，符合生物样品分析方法要求。药
动学研究结果显示，单次灌胃CLP后，大鼠血浆中原型CLP暴露量极低，其药-时曲线下面积（AUC0-35 h ）、峰浓度均小于或低于各代
谢物；活性代谢物CAM的AUC0-35 h约为CLP的43倍，但半衰期较短（2.53 h）；无活性代谢物CLP-C的暴露量最高，但达峰最迟且清
除缓慢；CLP-G的AUC0-35 h约为CAM的4倍，其半衰期与CLP-C近似。结论 本研究成功建立了测定CLP及其3种代谢产物含量
的液相色谱串联质谱法，并揭示了其在大鼠体内的药动学特征，即原型药物CLP可被迅速清除，而无活性代谢物CLP-C与CLP-G
均呈长半衰期特性，活性代谢物CAM呈短暂暴露模式。
关键词 氯吡格雷；代谢产物；液相色谱串联质谱法；药动学；血药浓度

Determination method of clopidogrel and its metabolites in rat plasma and its pharmacokinetic study
YI Huan，MIAO Lan，REN Changying，LIN Li，SUN Mingqian，PENG Qing，ZHANG Ying，LIU Jianxun（Beijing
Key Laboratory of Pharmacology of Chinese Materia Medica/Institute of Basic Medical Sciences of Xiyuan
Hospital， China Academy of Chinese Medical Sciences， Beijing 100091， China）

ABSTRACT OBJECTIVE To establish a method for determining the contents of clopidogrel （CLP）， clopidogrel carboxylate
（CLP-C）， clopidogrel acyl-β-D-glucuronide （CLP-G） and contents of clopidogrel active metabolite （CAM） in rat plasma， and to
investigate their in vivo pharmacokinetic characteristics. METHODS The Shisedo CAPCELL ADME column was used with a mobile
phase consisting of water and acetonitrile （both containing 0.1% formic acid） in a gradient elution. The flow rate was 0.4 mL/min，
and the column temperature was maintained at 20 ℃. The injection volume was 2 μL. The analysis was performed in positive ion
mode using electrospray ionization with multiple reaction monitoring. The ion pairs for quantitative analysis were m/z 322.1→211.9
（for CLP）， m/z 308.1→197.9 （for CLP-C）， m/z 322.1→154.8 （for CLP-G）， m/z 504.1→154.9 [for racemic CAM derivative
（CAMD）]. Six rats were administered a single intragastric dose of CLP （10 mg/kg）. Blood samples were collected before
medication and at 0.08， 0.33， 0.66， 1， 2， 4， 6， 10， 23 and 35 hours after medication. The established method was used to detect
the serum contents of various components in rats. Pharmacokinetic parameters were then calculated using WinNonlin 6.1 software.
RESULTS The linear ranges for CLP， CLP-C and CAMD were 0.08-20.00， 205.00-8 000.00， and 0.04-25.00 ng/mL， respectively
（r≥0.990）. The relative standard deviations for both intra-day and inter-day precision tests were all less than 15%， and the relative
errors for accuracy ranged from －11.68% to 14.40%. The coefficients of variation for the matrix factors were all less than 15%，
meeting the requirements for bioanalytical method validation. The results of the pharmacokinetic study revealed that， following a
single intagastric administration of CLP in rats， the exposure to the parent CLP in plasma was extremely low. Both the area under
the drug concentration-time curve （AUC0-35 h ） and the peak concentration of the parent CLP were lower than those of its metabolites.
The AUC0-35 h of the active metabolite CAM was approximately
Δ 基金项目 国家自然科学基金项目（No.81873179）；北京市研究 43 times that of CLP， though it had a shorter half-life （2.53
型病房卓越临床研究计划平行项目（No.BRWEP2024Z014170102）；中
h）. The inactive metabolite CLP-C exhibited the highest
国中医科学院科技创新工程项目（No.CI2021A04906） exposure level， but it reached its peak concentration the latest
＊第一作者 博士研究生。研究方向：药物代谢动力学。E-mail：
and was eliminated slowly. The AUC0-35 h of CLP-G was about
cathrine97@163.com
# 通信作者 教授，博士生导师，博士。研究方向：药物代谢动力 four times that of CAM， and its half-life was similar to that of
学。E-mail：zhyingde@sina.com CLP-C. CONCLUSIONS This study successfully established

中国药房 2025年第36卷第13期 China Pharmacy 2025 Vol. 36 No. 13 · 1599 ·