高效液相色谱法测定缬沙坦原料药中N-二甲基亚硝胺和N-二乙基亚硝胺的含量

Determination of N-nitrosodimethylamine and N-diethylnitrosamine in Valsartan by HPLC

目的 建立高效液相色谱法(HPLC)测定缬沙坦原料药中N-亚硝基二甲胺(NDMA)和N-亚硝基二乙胺(NDEA)的含量。方法 采用HPLC,色谱柱:Shim-pack Scepter C_(18)-120(4.6×250 mm,5μm),流动相A:甲醇-水(35∶65),流动相B:甲醇-水(75∶25)梯度洗脱的方式,流速:1.0 ml/min,柱温:30℃,进样量:20μl。结果 NDMA和NDEA在5~100 ng/ml浓度范围内线性关系良好,NDMA相关系数(r^(2))=0.999 8,平均回收率为90.8%~96.6%,相对标准偏差(RSD)为1.9%(n=9),检出限为1.5 ng/ml,定量限为5 ng/ml;NDEA相关系数(r2)=0.999 8,平均回收率为91.8%~99.8%,RSD为1.7%(n=9),检出限为1.5 ng/ml,定量限为5 ng/ml。结论 该操作简单、结果准确,可用于测定缬沙坦原料药中NDMA与NDEA的含量。

Objective To establish a method for the determination of N-nitrosodimethylamine(NDMA)and N-diethylnitrosamine(NDEA)in Valsartan.Methods Using HPLC,the chromatographic conditions were as follows:stationary phase:Shim-pack Scepter C_(18)-120(4.6 × 250 mm,5μm),mobile phase A:Methanol-Water(35:65),mobile phase B:Methanol-Water(75:25),flow rate:1.0 ml/min,column temperature:30℃,injection volume:20μl.Results The concentration of NDMA and NDEA showed a good linear relationship in the range of 5~100 ng/ml,the correlation coefficient of NDMA(r^(2))was 0.9998,the recovery was 90.8%~96.6%,RSD was 1.9%(n=9),the detection limit was 1.5 ng/ml,the quantitative limit was 5 ng/ml;the correlation coefficient of NNDEA(r^(2))was 0.9998,the recovery was 91.8%~99.8%,RSD was 1.7%,the detection limit was 1.5 ng/ml,the quantitative limit was 5 ng/ml.Conclusion The method is sensitive and accurate,and can be used for the determination of NDMA and NDEA in Valsartan.