| 引用本文: | 袁芳，陈卓佳，陈杰.HPLC测定大鼠血浆中盐酸川芎嗪及其代谢物[J].中国现代应用药学,2012,29(6):529-534. |
| YUAN Fang, CHEN Zhuojia, CHEN Jie.Determination of Tetramethylpyrazine and its Active Metabolite in Rat Plasma by HPLC[J].Chin J Mod Appl Pharm(中国现代应用药学),2012,29(6):529-534. |
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| 摘要: |
| 目的 建立川芎嗪(TMP)及其代谢物2-羟甲基-3，5，6-三甲基吡嗪(HTMP)大鼠血浆中的高效液相色谱测定方法。方法 血浆碱化后用氯仿-1-氯正丁烷(3∶1)萃取，采用HPLC检测，色谱柱为Hypersil BDS C18(4.6 mm×250 mm，5 mm)，流动相为20 mmol·L-1磷酸二氢钾缓冲液(pH 5.6)-甲醇(72∶28)，2-甲基吡嗪为内标，流速：1.0 mL·min-1。检测波长为285 nm，柱温为室温。结果 TMP大鼠血浆浓度测定方法在0.031 25~50 μg·mL-1内线性良好(r=0.999 5)；HTMP线性范围为0.031 25~5 μg·mL-1(r=0.999 8)。TMP、HTMP的定量下限均为0.031 25 μg·mL-1，提取回收率为86.2%~93.0%(TMP)、73.8%~95.1%(HTMP)，相对回收率为96.9%~117.7%(TMP)、97.5%~104.9%(HTMP)，批内和批间RSD均<9%(n=5)。结论 该方法简便、准确，重复性好，灵敏度高，适合于TMP及其主要代谢产物HTMP血药浓度测定及药动学研究。 |
| 关键词: 川芎嗪 血药浓度 高效液相色谱法 |
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| 基金项目:广东省自然科学基金(8451008901000788)；广东省医学科研基金(A2009163) |
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| Determination of Tetramethylpyrazine and its Active Metabolite in Rat Plasma by HPLC |
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YUAN Fang, CHEN Zhuojia, CHEN Jie1,2,3
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1.School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China;2.Institute of Clinical Pharmacology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China;3.Department of Pharmacy, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510006, China
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| Abstract: |
| OBJECTIVE To establish an HPLC method for the determination of concentration of tetramethylpyrazine (TMP) and its active metabolite, 2-hydroxymethyl-3,5,6-trimethylpyrarzine (HTMP) in rat plasma and apply this method to the pharmacokinetic study of TMP. METHODS 2-Methylpyrazine was used as internal standard. The alkalified serum samples were extracted with a chloroform-1-chloro-butane (3∶1) induced liquid-liquid extraction. The target analytes were quantitatively determined by HPLC. Hypersil BDS C18 column(4.6 mm×250 mm, 5 mm) was used. The mobile phase consisted of 20 mmol·L-1 potassium dihydrogen phosphate buffer(pH 5.6)-methanol(72∶28). The flow rate was 1.0 mL·min-1, 15 mL sample was injected and detected by the ultra-violet detector at 285 nm. RESULTS The linearity was obtained over the concentration ranges of 0.031 25-50 μg·mL-1 for TMP(r=0.999 5) and 0.031 25-5 μg·mL-1 for HTMP(r=0.999 8). The lower limit of quantitation (LLOQ) was 0.031 25 μg·mL-1 for TMP and HTMP. The inter- and intra-batch precisions (RSD%) were less than 9% for both analyses. The accuracies and extracted recoveries were 86.2%-93.0% for TMP and 73.8%-95.1% for HTMP. The relative recoveries were 96.9%-117.7% for TMP and 97.5%-104.9% for HTMP. CONCLUSION The method has good selectivity, acceptable accuracy, precision and sensitivity. It can be applied to pharmacokinetic study of TMP in rats. |
| Key words: tetramethylpyrazine serum concentration of drug HPLC |
