RP-HPLC法测定血清中双环醇的含量

Determination of bicyclol in human serum by RP-HPLC

目的 建立反相高效液相色谱快速检测双环醇血药浓度的方法。方法 用 10 %高氯酸和 2 0 %三氯乙酸直接沉淀血清蛋白 ,三氯甲烷抽提血清杂质 ,离心后取上清液进行色谱分析。色谱柱为Supecol C18(5 μm ,4 6mm× 2 5 0mm)不锈钢柱 ,流动相为甲醇 -水 -乙腈 (5 1∶4 3∶6 ) ,流速 1 2ml·min-1,紫外检测波长为 2 5 4nm ,进样量 2 0 μl,柱温 2 2℃ ,以联苯双脂为内标。 结果 血清药物浓度在 0 1～ 4 0mg·L-1峰面积与浓度呈良好的线性 (r =0 9985 ) ,色谱峰分离良好 ,无干扰 ,平均回收率大于97 5 9% (n =5 ) ,日内RSD <4 % ,日间RSD <6 % (n =6 )。结论 该法简便、快速、定量准确 。

AIM To establish a RP HPLC method for determination of bicyclol in human serum. METHODS\ The serum protein was precipitated by addition of 10% perchloric acid and 20% trichloracetic acid. Chloroform was used to extract the interfering materials in the serum. The resulting supernatant liquid was directly injected after centrifugation. The chromatography condition was:Supecol C 18 ODS(5 μm, 4 6 mm×250 mm) stainless steel column, methanol water acetonitrile(51∶43∶6) as mobile phase at 1 2 ml·min -1 . Detection wave length was 254 nm, injection volume was 20 μl. bifendate was used as the internal standard. RESULTS\ The chromatography was good and not interfered by the components of the serum. The linear range of bicyclol was within drugserum concentrations of 0 1～4 0 g·L -1 ( r = 0 998 5 ). The recovery was more than 97 59%( n =5). The intra day RSD was less than 4% and the inter day RSD was less than 6%( n =5). CONCLUSION\ This method is simple, rapid and accurate. It is applied to study the pharmacokinetic and bioavailability of bicyclol in human serum.