摘要
目的:建立高效液相色谱法测定盐酸左旋咪唑片的有关物质及含量.方法:有关物质检测采用Ther-mo Hypsil C18(100 mm ×4.6 mm,3 μm)色谱柱,以0.75％磷酸二氢铵溶液(用二异丙胺调节pH至7.0)-乙腈为流动相,梯度洗脱;含量测定采用InertSustain C18(150 mm ×4.6 mm,5 μm)色谱柱,以0.75％磷酸二氢铵溶液(用二异丙胺调节pH至7.0)-乙腈(70∶30)为流动相.流速均为1.0mL·min-1,检测波长均为215 nm,进样体积均为10 μL.结果:建立的有关物质检测方法能较好地分离盐酸左旋咪唑与5个已知杂质;已知杂质 A、B、C、D、E 质量浓度在 10.87～25.37、11.62～27.10、12.38～28.90、30.89～72.07、12.41～28.95 μg·mL-1的范围内与峰面积呈良好的线性关系(r＞0.999),平均校正因子分别为1.6、1.4、2.6、1.2、2.4,平均回收率(n=9)分别为 98.1％、99.0％、98.6％、100.1％和 99.9％,RSD 分别为 0.63％、0.79％、0.92％、0.96％、0.33％.建立的含量测定方法能分离盐酸左旋咪唑与相邻杂质C;盐酸左旋咪唑质量浓度在0.101 4～0.304 3 mg·mL-1的范围内与峰面积呈良好的线性关系(r=0.999 9);盐酸左旋咪唑的平均回收率(n=9)为100.2％,RSD=0.52％;盐酸左旋咪唑的定量限与检测限分别为0.076 ng和0.030 ng.采用建立的方法实际测定5家企业生产的10批样品,总杂质量为0.05％～0.62％,含量为99.5％～104.3％.结论:研究建立的盐酸左旋咪唑片的有关物质和含量测定的方法专属性强,结果准确度高,可用于盐酸左旋咪唑片中杂质的控制研究以及质量标准的提高.
Abstract
Objective:To develop an HPLC method for the determination of related substances and assay of le-vamisole hydrochloride tablets.Methods:The chromatographic separation of related substances was performed on a Thermo Hypsil C18 column(100 mm ×4.6 mm,3 μm).A gradient elution was applied with a mobile phase composed of0.75％monobasic ammonium phosphate solution(adjusted at pH 6.5 with diiopropylamine)and ace-tonitrile.The assay analytical column was packed with IntertSustain C18column(150 mm ×4.6 mm,5 μm).The mobile phase was 70 volumes of 0.75％monobasic ammonium phosphate solution(adjusted at pH 7.0 with diio-propylamine)with 30 volumes of acetonitrile.The wavelength detection was set at 215 nm,the injection volumn was 10 μL,and the flow rate was 1.0 mL·min-1.Results:Levamisole and its impurities were separated well by related substances HPLC method above.Impurities A,B,C,D,E showed the good linearities in the concentra-tion ranges of 10.87-25.37 μg·mL-1,11.62-27.10 μg·mL-1,12.38-28.90 μg·mL-1,30.89-72.07μg·mL-1,12.41-28.95 μg·mL-1(r＞0.999).The average correction factors of impurities A,B,C,D,E determined by three columns and liquid chromatographies were 1.6,1.4,2.6,1.2,2.4,respectively,the recovery rates(n=9)were 98.1％,99.0％,98.6％,100.1％,99.9％and the RSDs were 0.63％,0.79％,0.92％,0.96％,0.33％.The separation of levamisole and impurity C was good by assay HPLC method.Levamisole showed the good linearity in the concentration range of 0.10 14-0.304 3 mg·mL-1(r=0.999 9).The average recovery rate(n=9)of levamisole was 100.2％,RSD=0.52％.Ten batches levamisole hydrochlo-ride tablets from five domestic pharmaceutical enterprises were determined by the above related substances and assay HPLC method,the total impurities mass were 0.05％-0.62％,and the assays were 99.5％-104.3％.Conclusion:The established method is high selection and accurate.It is suitable applied for determination of related substances and assay of levamiole hydrochloride tablets.
基金项目
国家药典委员会药品标准提高课题(2016)(化175)
NETL
NSTL
万方数据