返回 
加入收藏
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
x
请在关注微信后，向客服人员索取文件
| 篇名： | 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 |
| TITLE： | Determination method of clopidogrel and its metabolites in rat plasma and its pharmacokinetic study |
| 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ShisedoCAPCELLADME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0.4mL/min，柱温为20℃，进样量为2μL；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/z322.1→211.9（CLP）、m/z308.1→197.9（CLP-C）、m/z322.1→154.8（CLP-G）、m/z504.1→154.9[外消旋CAM衍生物（CAMD）]。取6只大鼠单次灌胃CLP（10mg/kg），分别于给药前和给药后0.08、0.33、0.66、1、2、4、6、10、23、35h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WinNonlin6.1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、205.00～8000.00、0.04～25.00ng/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（AUC0-35h）、峰浓度均小于或低于各代谢物；活性代谢物CAM的AUC0-35h约为CLP的43倍，但半衰期较短（2.53h）；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AUC0-35h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。 |
| 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 43 times that of CLP， though it had a shorter half-life （2.53 h）. The inactive metabolite CLP-C exhibited the highest exposure level， but it reached its peak concentration the latest and was eliminated slowly. The AUC0-35 h of CLP-G was about four times that of CAM， and its half-life was similar to that of CLP-C. CONCLUSIONS This study successfully established an liquid chromatography-tandem mass spectrometry method for the determination of CLP and its three metabolites， and revealed their pharmacokinetic characteristics in rats. Specifically， the parent drug CLP was rapidly eliminated， while the inactive metabolites CLP-C and CLP-G exhibited long half-lives， and active metabolite CAM displayed a transient exposure pattern. |
| 期刊： | 2025年第36卷第13期 |
| 作者： | 易欢；苗兰；任常英；林力；孙明谦；彭勍；张颖；刘建勋 |
| AUTHORS： | YI Huan，MIAO Lan，REN Changying，LIN Li，SUN Mingqian，PENG Qing，ZHANG Ying，LIU Jianxun |
| 关键字： | 氯吡格雷；代谢产物；液相色谱串联质谱法；药动学；血药浓度 |
| KEYWORDS： | clopidogrel； metabolites； liquid chromatography-tandem mass spectrometry method； pharmacokinetics； plasma |
| 阅读数： | 0 次 |
| 本月下载数： | 0 次 |
* 注：未经本站明确许可，任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作！
