HPLC测定大鼠微透析液和血浆中苦参碱的浓度

Determination of Martrine in Rat Microdialysate and Plasma by HPLC

关键词:  微透析  肝  血  血浆  苦参碱  高效液相色谱

Key words:  microdialysis  liver  blood  plasma  matrine  HPLC

摘要:

目的 建立大鼠微透析液和血浆中苦参碱的反相高效液相检测方法。方法 色谱柱为Diamonsil C₁₈柱(4.6 mm×150 mm，5 μm)；流动相为乙腈-0.1%磷酸(用三乙胺调pH至7.6~7.7)(微透析液测定：35∶65；血浆测定：30∶70)；流速1.0 mL·min^-1；检测波长220 nm；柱温35 ℃；进样量20 μL。结果 本方法测定苦参碱线性范围微透析液为0.10~25.00 μg·mL^-1，血浆为0.32~81.80 μg·mL^-1，相关系数均为1。微透析液的日内、日间精密度RSD≤6.95 %，血浆RSD≤10.75 %。微透析液的平均回收率为94.7~105.9 %，血浆为98.6~107.9 %，稳定性良好。结论 本方法准确、简单、专属性强，适用于苦参碱大鼠体内药动学研究中微透析液和血浆中药物浓度的测定。

Abstract:

OBJECTIVE To develop a sensitive and specific reverse-phase high-performance liquid chromatoraphy (RP-HPLC) method to determine matrine in rat microdialysate and plasma. METHODS The samples were analyzed on a Diamonsil C₁₈ column(4.6 mm×150 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.1% phosphoric acid(adjusted pH to 7.6-7.7 with triethylamine ) (35∶65 in microdialysate and 30∶70 in plasma). Flow rate was 1.0 ml·min^-1. The detection wavelength was 220 nm. The column temperature was 35 ℃. The injection volume was 20 μL. RESULTS The assay showed good linear correlation over the range of 0.10-25.00 μg·mL^-1 in microdialysate and 0.32-81.80 μg·mL^-1 in plasma(r=1, n=5) , respectively. The intra- and inter-day precision over these ranges were not more than 6.95 % in microdialysate and 10.75 % in plasma, respectively. The average accuracy was in the range of 94.7-105.9 % in microdialysate and 98.6-107.9 % in plasma, respectively. The sample solution was stable. CONCLUSION The method is accurate, simple, sensitive and specific for determination of matrine in rat microdialysate and plasma.