高效液相色谱法测定复方胰酶片中胆酸和牛磺猪去氧胆酸的含量

Determination of Cholic Acid and Taurohyodeoxycholic Acid in Compound Pancreatin Tablets by HPLC

建立了高效液相色谱法测定复方胰酶片中胆酸和牛磺猪去氧胆酸含量的方法。取复方胰酶片10片,研细,分别称取样品0.3,0.5 g,加甲醇溶解,超声提取30 min,用甲醇分别定容至50,25 mL。所得溶液以SunShell HFC_(18)-30色谱柱为固定相分离胆酸,以Diamonsil C_(18)色谱柱为固定相分离牛磺猪去氧胆酸,流动相为体积比35∶65的乙腈-含0.1%(体积分数)磷酸溶液的混合液,检测波长为192 nm。在优化的试验条件下,胆酸和牛磺猪去氧胆酸的质量浓度在一定范围内与其对应的峰面积呈线性关系,检出限(3S/N)分别为11.0,5.61 mg·L^(-1)。按标准加入法进行回收试验,回收率为91.9%和102%,其测定值的相对标准偏差(n=6)均小于5.0%。

A method was established for the determination of cholic acid and taurohyodeoxycholic acid in compound pancreatin tablets by HPLC.The samples(0.3 g and 0.5 g)were respectively taken out of 10 ground compound pancreatin tablets,dissolved with methanol,and extracted ultrasonically for 30 min.The extracts were made their volume up to 50,25 mL with methanol respectively.The obtained solution was separated on SunShell HFC_(18)-30 column for cholic acid,and Diamonsil C_(18) column for taurohyodeoxycholic acid.A mixture of acetonitrile and 0.1%(φ)phosphoric acid solution at a volume ratio of 35∶65 was used as the mobile phase,and the detection wavelength was 192 nm.Under the optimized test conditions,linear relationships between values of peak area and mass concentration of cholic acid and taurohyodeoxycholic acid were found in definite ranges,with detection limits(3 S/N)of 11.0 mg·L^(-1) and 5.61 mg·L^(-1),respectively.Test for recovery was made by standard addition method,giving results of 91.9%and 102%,and RSDs(n=6)of the determined values less than 5.0%.