RP-HPLC测定老鼠血浆中芦丁的含量

HPLC Determination of Rutin in Mouse Plasma

目的:建立老鼠血浆中芦丁含量的HPLC测定方法。方法:采用Nova-Pak C18色谱柱(200 mm×4.6mm,5 μm)及BondapakC18guard-pak cartridge保护柱;乙腈-10 mmol·L-1醋酸铵(含0.3 mmol·L-1EDTA)-冰醋酸(15:83:2)为流动相,用前经0.45 μm的过滤膜,超声脱气;流速为1.0 mL·min-1;紫外检测波长为260nm;柱温为30℃,进样量20μL。结果:最低检测限为0.5 ng,线性范围0.05-100μg·mL-1,平均回收率在96.43%以上,日内及日间精密度的相对标准差分别小于3%和5%。结论:该方法简单,快速,灵敏度高,重复性好。

Objective:To establish the method for determining rutin in mouse plasma by reversed phase high performance liquid chromatography. Methods: Rutin was successfully resolved using a Nova -Pak C18 column (200 mm ×4. 6 mm,5μm) and Bondapak C18guard - pak cartridge. Mobile phase consisting of acetonitrile - 10 mmol·L-1 ammonium acetate solution containing 0. 3 mmol·L-1 EDTA - acetic acid( 15: 83: 2) were degassed ultrasonically before use. Each component of the mobile phase was filtered through a 0. 45μm membrane. All separation were at 30℃and the flow - rate was 1. 0 mL·min -1. Injection volume was 20μL. The peak of rutin was monitored to set at UV 260 nm. Results:The detection limit was 0. 5 ng. The linear range was 0.05 - 100 μg·mL-1 . The average recovery was over 96. 43% . RSD of intra - day and inter - day were less than 3% and 5% ,respectively. Conclusion:The method is simple, rapid, sensitive and producible.