HPLC法测定硫辛酸注射液中的有关物质

Determination of Related Substances in Thioctic Acid Injection by HPLC

目的：建立测定硫辛酸注射液中有关物质的方法。方法：采用高效液相色谱法。色谱柱为Agilent Eclipse Plus C18,流动相A为0.005 mol/L磷酸二氢钾溶液（用磷酸调节p H至3.0～3.1）、流动相B为甲醇-乙腈（1∶1,V/V）（梯度洗脱）,流速为1.2 ml/min,检测波长为215 nm,柱温为35℃,进样量为20μl。结果：在所建立的色谱条件下,硫辛酸与各杂质及其降解产物分离良好;硫辛酸检测质量浓度线性范围为0.828～52.992μg/ml（r=0.999 9）;精密度、稳定性试验的RSD〈3%;杂质A的加样回收率为98.45%～105.73%,RSD=2.41%（n=9）;其最大未知单个杂质的质量分数为0.152%～0.189%,总杂质的质量分数为0.487%～0.516%。结论：该方法简便、准确、专属性强、灵敏度高,可用于硫辛酸注射液中有关物质的测定。

OBJECTIVE：To establish a method for the determination of related substances in Thioctic acid injection. METHODS：HPLC was performed on the column of Agilent Eclipse Plus C18 with mobile phase A of 0.005 mol/L potassium dehydrogenate phosphate solution（adjusted with phosphoric acid to p H 3.0 to 3.1）and B of methanol：aoetonitrile（1∶1,V/V）（gradient elution）at a flow rate of 1.2 ml/min;detection wavelength was 215 nm,column temperature was 35 ℃,and the injection volume was 20 μl. RESULTS：Under the established chromatographic conditions,thioctic acid with impurities and its degradation products were well separated;the linear range of thioctic acid was 0.828-52.992 μg/ml（r=0.999 9）;RSDs of precision and stability tests were lower than 3%;recovery of impurity A was 98.45%-105.73%（RSD=2.41%,n=9）;the largest single impurity content in mass fraction was 0.152%-0.189%,the total impurity content in mass fraction was 0.487%-0.516%. CONCLUSIONS：The method is simple,accurate,specific and sensitivity,and can be used to determine the related substances of the Thioctic acid injection.