Determination of related substances and assay of levamisole hydrochloride tablets by HPLC
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.
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