RP-HPLC法测定熊去氧胆酸胶囊中熊去氧胆酸及有关物质的含量

目的建立反相高效液相色谱法(RP-HPLC)测定熊去氧胆酸胶囊中熊去氧胆酸、胆酸、鹅去氧胆酸及胆石酸的含量。方法使用Diamonsil C18色谱柱(200 mm×4.6 mm,5μm),流动相为乙腈-0.03 mol·L-1磷酸溶液(体积比为40∶60),流速为1.4 mL·min-1,检测波长为205 nm,柱温为35℃。结果熊去氧胆酸胶囊中熊去氧胆酸、胆酸、鹅去氧胆酸及胆石酸在25 min内洗脱并基线分离。熊去氧胆酸、胆酸、鹅去氧胆酸及胆石酸的线性范围分别为0.80²00.00、0.45200.00、0.45¹10.00、0.30110.00、0.30⁷0.00和0.3070.00和0.30⁸0.00 mg·L-1,平均回收率分别为99.7%、99.3%、98.7%和99.1%,RSD分别为1.30%、1.47%、1.87%和1.95%(n=3)。结论 RP-HPLC可用于同时测定熊去氧胆酸胶囊中熊去氧胆酸、胆酸、鹅去氧胆酸及胆石酸的含量,可应用于熊去氧胆酸胶囊胶囊制剂的质量控制。

Determination of ursodeoxycholic acid,chenodeoxycholic acid,cholic acid and lithocholic acid in compound ursodeoxycholic acid capsules by RP-HPLC

Objective To establish a reversed phase high performance liquid chromatography (RP - HPLC) to determine ursodeoxycholic acid, chenodeoxycholic acid, cholic acid and lithocholic acid in compound ursodeoxycholic acid capsules. Methods Diamonsil C₁₈ chromatographic column (200 mm×4.6 mm, 5 um) was used in the study, mobile phase was acetonitrile-0.03 mol·L^-1 of phosphate solution(V:V=40:60), the flow rate was 1.4 mL·min^{- 1}, the detection wavelength was set at 205 nm, the column temperature was 35 c. Results Ursodeoxycholic acid, chenodeoxycholic acid, cholic acid and lithocholic acid was completely separated within 25 min. A good linear range of 0.80-200.00 mg•L^-1 for ursodeoxycholic acid, 0.45-110.00 mg•L^-1 for cholic acid, 0.30-70.00 mg•L^-1 for chenodeoxycholic acid and 0.30-80.00 mg•L^-1 for lithocholic acid were showed respectively. The average recoveries of ursodeoxycholic acid, cholic acid, chenodeoxycholic acid and lithocholic acid were 99.7%(RSD=1.30%), 99.3％(RSD=1.47%), 98.7% (RSD=1.87%) and 99.1% (RSD=1.30%) respectively (n=12). Conclusions This method can simultaneously determine ursodeoxycholic acid, chenodeoxycholic acid, cholic acid and lithocholic acid contents in compound ursodeoxycholic acid capsules. The method is simple, accurate and reliable, and can be applied for the quality control of the compound ursodeoxycholic acid capsule.