摘要
目的:建立3α-羟基-7-氧代-胆烷酸有关物质的分析方法。方法:以十八烷基硅烷键合硅胶为填充剂,以0.05%的磷酸水溶液和乙腈为流动相梯度洗脱,流速为1.0 mL·min^(-1),检测波长为202 nm。结果:3α-羟基-7-氧代-胆烷酸样品溶液的色谱图中主峰与已知杂质之间和各已知杂质之间均能有效分离,3α-羟基-7-氧代-胆烷酸、杂质01、杂质04、杂质05和杂质06-A分别在1.00~24.97μg·mL^(-1)(r=0.999 9),1.00~50.05μg·mL^(-1)(r=0.999 9),0.75~25.04μg·mL^(-1)(r=0.999 9),1.01~5.02μg·mL^(-1)(r=0.999 8),1.00~9.98μg·mL^(-1)(r=0.999 9)的浓度范围内与峰面积成良好的线性关系;检测限分别为12.5、12.5、10、12.5、12.5 ng。结论:经验证,该方法简单易行,且专属性、灵敏度、线性和精密度良好,适用于本品有关物质的测定。
Objective： To establish a test method of related substances for 3or- hydroxy-7- oxo-cholanic acid. Methods：The test was performed on the octadecylsilane chemically bonded silica column with the gradient elution of 0. 05 % phosphoric acid aqueous solution and acetonitrile. The flow rate was 1.0 mL· min ^-1, and the detection wavelength was 202 nm. Results：There were good resolutions between the peak of 3α- hydroxy-7- oxo-cholanic acid and identified impurity peak, the different identified impurities peaks in the chromatogram of the test solution. There were good linear relationship between the peak area and concentration in the ranges of 1.00 -24. 97μg·mL^-1（r=0.9999）, 1.00-50.05μg·mL^-1（r=0.999 9）, 0.75-25.04 μg·mL^-1（r=0.999 9）, 1.01 - 5.02 μg·mL^-1（ r = 0. 999 8 ）, 1.00 - 9. 98μg·mL^-1 （ r = 0. 999 9 ） for 3or- hydroxy-7- oxo-cholanic acid, impurities 01,04, 05, 06-A respectively. The limits of detection for 3ct- hydroxy-7- oxo-eholanic acid, im- purities 01, 04, 05, and 06-A were 12. 5, 12.5, 10, 12. 5 and 12. 5 ng respectively. Conclusion： The validation shows that this simple and feasible method has the good specificity, sensitivity, linearity and precision. It is suit- able for the determination of related substances for 3α- hydroxy-7- oxo-cholanic acid.
出处
《中国药品标准》
CAS
2017年第4期283-287,共5页
Drug Standards of China
