HPLC加校正因子的主成分自身对照法测定氢溴酸高乌甲素注射液中2个有关物质的含量  

Determination of two related substances in lappaconitine hydrobromide injection by HPLC-principal component self-compare with correction factor

作　　者：徐树娟 张磊 马淑凤 付霞 卢娜 吴传立 XU Shu-juan;ZHANG Lei;MA Shu-feng;FU Xia;LU Na;WU Chuan-li(Linyi Inspection and Testing Center,Linyi 276000,China)

机构地区：[1]临沂市检验检测中心,临沂276000

出　　处：《药物分析杂志》2024年第11期1967-1974,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：2023年山东省药品质量风险监测工作专项(编号:鲁食药检业[2023]187号)。

摘　　要：目的:建立HPLC加校正因子的主成分自身对照法测定氢溴酸高乌甲素注射液中2个有关物质(N-去乙酰高乌甲素和冉乌头碱)的含量。方法:采用Kromasil 300-5-C18(250 mm×4.6 mm,5μm)色谱柱,以0.04 mol·L^(-1)磷酸二氢钾溶液-甲醇-乙腈(68∶17∶15)为流动相,检测波长252 nm,流速0.8 mL·min^(-1),柱温37℃,进样体积10μL。绘制氢溴酸高乌甲素和2个杂质的线性方程,分别以斜率计算各杂质相对于氢溴酸高乌甲素的校正因子,用相对保留时间确定各杂质的位置。测定4家企业共25批氢溴酸高乌甲素注射液中2个杂质含量,并与外标法测得结果进行比较。结果:本方法能较好地分离氢溴酸高乌甲素与2个杂质。N-去乙酰高乌甲素和冉乌头碱的相对保留时间分别为1.20和1.39,校正因子分别为1.23和0.94。氢溴酸高乌甲素、N-去乙酰高乌甲素和冉乌头碱质量浓度分别在0.9517~38.07、1.047~41.87和1.001~40.02μg·mL^(-1)的范围内与峰面积呈良好的线性关系(r均为1.000),平均回收率分别为100.2%、100.5%和100.5%,RSD均小于2.0%,检测限分别为0.095、0.10和0.10μg·mL^(-1),定量限分别为0.32、0.35和0.33μg·mL^(-1)。加校正因子的主成分自身对照法测得25批氢溴酸高乌甲素注射液中N-去乙酰高乌甲素的含量为0.31%~0.82%,冉乌头碱的含量为0~0.09%,与外标法测定结果一致。结论:经方法学验证,所建立的方法简便、快速,可准确测定氢溴酸高乌甲素注射液中有关物质的含量。Objective:To establish an HPLC-principal component self-compare with correction factor method for quantification of two related substances(N-deacetyllappaconitine and ranaconitine)in lappaconitine hydrobromide injection.Methods:The analysis was performed on a Kromasil 300-5-C_(18)(250 mm×4.6 mm,5μm)column with a mobile phase composed of 0.04 mol·L^(-1)potassium dihydrogen phosphate solution,methanol and acetonitrile(68∶17∶15).The detection wavelength was 252 nm,the flow rate was 0.8 mL·min^(-1),the column temperature was 37℃,and the injection volume was 10μL.The slope of linear equation was used to determine the relative correction factor between the two impurities and lappaconitine hydrobromide.The relative retention time was used to determine the position of related substances.The contents of two impurities in 25 batches of lappaconitine hydrobromide injection produced by four pharmaceutical companies were determined and compared with the results of the external standard method.Results:Lappaconitine hydrobromide and the impurities were separated well by this method.The relative retention time of N-deacetyllappaconitine and ranaconitine were 1.20 and 1.39,and the correction factors were 1.23 and 0.94,respectively.Lappaconitine hydrobromide,N-deacetyllappaconitine and ranaconitine showed good linearity in the mass concentration ranges of 0.9517-38.07μg·mL^(-1),1.047-41.87μg·mL^(-1)and 1.001-40.02μg·mL^(-1)with r=1.000,respectively.The average recovery rates of lappaconitine hydrobromide,N-deacetyllappaconitine and ranaconitine were 100.2%,100.5%and 100.5%respectively,with RSD less than 2.0%.The limits of detection(LOD)of lappaconitine hydrobromide,N-deacetyllappaconitine and ranaconitine were 0.095,0.10,0.10μg·mL^(-1)and the limits of quantitation(LOQ)were 0.32,0.35 and 0.33μg·mL^(-1),respectively.The content of N-deacetyllappaconitine in 25 batches of lappaconitine hydrobromide injection was in the range of 0.31%-0.82%and the content of ranaconitine was in the range of 0%-0.09%.It was consis

关 键 词：氢溴酸高乌甲素注射液 高效液相色谱法 加校正因子的主成分自身对照法 有关物质 N-去乙酰高乌甲素 冉乌头碱 

分 类 号：R917[医药卫生—药物分析学]