人工牛黄甲硝唑胶囊中主要胆汁酸类成分的HPLC-ELSD法测定研究

目的: 建立同时测定人工牛黄甲硝唑胶囊中来源于原料人工牛黄中的5种胆汁酸类成分(牛磺胆酸钠、胆酸、猪去氧胆酸、鹅去氧胆酸和去氧胆酸)的方法。方法: 采用高效液相色谱仪与蒸发光散射检测器联用技术对人工牛黄甲硝唑胶囊进行测定。色谱条件:Agilent TC-C₁₈色谱柱(4.6 mm×250 mm,5 μm),柱温35 ℃;流动相为0.2%甲酸水溶液(A)-乙腈(B),梯度洗脱(0~10 min,15%B→32%B;10~15 min,32%B→50%B;15~22 min,50%B→68%B;22~23 min,68%B→95%B;23~28 min,95%B),流速1.0 mL·min^-1;ELSD检测器:氮气流速2.3 L·min^-1,漂移管温度115 ℃。结果: 牛磺胆酸钠、胆酸、猪去氧胆酸、鹅去氧胆酸、去氧胆酸质量浓度分别在24.93~249.27、47.62~476.19、39.72-397.23、6.29~62.92和10.63~106.28 μg·mL^-1范围内与峰面积成良好对数线性关系;平均回收率(n=6)分别为99.9%(RSD=1.6%)、98.9%(RSD=1.6%)、99.2%(RSD=2.0%)、99.4%(RSD=2.0%)和98.6%(RSD=2.0%);3批样品测定结果分别为0.16%~0.37%、0.67%~0.75%、0.44%~0.70%、0.10%~0.17%、0.16%~0.19%。结论: 该法可同时测定人工牛黄甲硝唑胶囊中来源于原料人工牛黄的5种胆汁酸类成分(牛磺胆酸钠、胆酸、猪去氧胆酸、鹅去氧胆酸和去氧胆酸)的含量,适用于人工牛黄甲硝唑胶囊的质量评价与控制。

Objective: To establish a quantitative method for simultaneous determination of 5 bile acids (sodium taurocholate,cholic acid,hyodeoxycholic acid,chenodeoxycholic acid,deoxycholic acid)in artificial bezoar and metronidazole capsules. Methods: The HPLC-ELSD method was performed using an Agilent TC-C₁₈ column (4.6 mm×250 mm,5 μm)with a column temperature of 35 ℃ and a liquid flow-rate of 1.0 mL·min^-1 using 0.2% aqueous formic acid (A)and acetonitrile (B)as the mobile phase with a linear gradient (0-10 min,15%B→32%B;10-15 min,32%B→50%B;15-22 min,50%B→68%B;22-23 min,68%B→95%B;23-28 min,95%B).An evaporate light-scattering detector (ELSD)was used as the detector with a nitrogen flow rate of 2.3 L·min^-1,at a drift tube temperature of 115 ℃. Results: The linear ranges of sodium taurocholate,cholic acid,hyodeoxycholic acid,chenodeoxycholic acid and deoxycholic acid were 24.93-249.27 μg·mL^-1,47.62-476.19 μg·mL^-1,39.72-397.23 μg·mL^-1,6.29-62.92 μg·mL^-1 and 10.63-106.28 μg·mL^-1,respectively.The corresponding average recovery rates (n=6)were 99.9%,98.9%,99.2%,99.4% and 98.6%;the relative standard deviations were 1.6%,1.6%,2.0%,2.0%,2.0%,respectively.The results of sample determination were 0.16%-0.37%,0.67%-0.75%,0.44%-0.70%,0.10%-0.17% and 0.16%-0.19%,respectively. Conclusion: The method can simultaneously determine 5 bile acids (sodium taurocholate,cholic acid,hyodeoxycholic acid,chenodeoxycholic acid,deoxycholic acid)in artificial bezoar and metronidazole capsules,which can be used for the evaluation and quality control of artificial bezoar and metronidazole capsules.