HPLC法梯度洗脱同时测定硫辛酸注射液有关物质、苯甲醇和苯甲醛

Simultaneous Determination of the Related Substances,Benzyl Alcohol and Benzaldehyde in Alpha Lipoic Acid Injection by HPLC with Gradient Elution

目的:建立梯度洗脱的HPLC法,以双波长同时测定硫辛酸注射液的有关物质、苯甲醇及其降解产物苯甲醛。方法:采用Shiseido MGⅡC18(250 mm×4.6 mm,5μm)色谱柱,以0.005 mol·L^-1磷酸二氢钾溶液-甲醇-乙腈(70∶26∶4)(用磷酸调pH至3.0)为流动相A,以0.005 mol·L^-1磷酸二氢钾溶液-甲醇-乙腈(41∶51∶8)(用磷酸调pH至3.0)为流动相B,梯度洗脱,流速为1.5 ml·min^-1,检测波长为250 nm和215 nm,柱温为30℃,进样量为20μl。结果:本法可有效分离硫辛酸注射液中各色谱峰。硫辛酸、苯甲醇和苯甲醛浓度分别在1~50μg·ml^-1(r=0.9999)、40~2000μg·ml-1(r=1.0000)和0.04~2μg·ml^-1(r=1.0000)范围内与峰面积线性关系良好;平均回收率分别为100.2%(RSD=0.6%),99.9%(RSD=0.4%)和99.2%(RSD=1.3%)(n=9);检测限(LOD)分别为0.38,7,0.02μg·ml-1。结论:本方法专属、灵敏,可用于硫辛酸注射液的质量控制。

Objective:To develop an HPLC with gradient elution method for the simultaneous determination of the related substances,benzyl alcohol and benzaldehyde in alpha lipoic acid injection.Methods:The samples were separated by HPLC with gradient elution on a Shiseido MGⅡC18(250 mm×4.6 mm,5μm)column with two mobile phases of phosphate buffer-methanol-acetonitrile(adjusting pH to 3.0 with phosphoric acid,70∶26∶4)(A)and phosphate buffer-methanol-acetonitrile(adjusting pH to 3.0 with phosphoric acid,41∶51∶8)(B).The flow rate was 1.5 ml·min^-1.The detection wavelengths were 250 nm and 215 nm.The column temperature was 30℃.The injection volume was 20μl.Results:Good separation of the related substances was achieved.The calibration curve for alpha lipoic acid,benzyl alcohol and benzaldehyde was linear over the range of 1-50μg·ml^-1(r=0.9999),40-2000μg·ml^-1(r=1.0000)and 0.04-2μg·ml^-1(r=1.0000),respectively.The average recovery was 100.2%(RSD=0.6%),99.9%(RSD=0.4%)and 99.2%(RSD=1.3%)(n=9),respectively.The LOD was 0.38μg·ml-1,7μg·ml-1 and 0.02μg·ml-1,respectively.Conclusion:The assay can be applied to the quality control of alpha lipoic acid injection.