盐酸伊伐布雷定中卤代烷烃类潜在遗传毒性杂质检测方法的建立

Establishment of one method for determination of haloalkane genotoxic impurities in ivabradine hydrochloride

目的首次建立了一种用于盐酸伊伐布雷定原料药中卤代烷烃类遗传毒性杂质测定的方法。方法使用CAPCELL PAK MGⅡC18(150 mm×4.6 mm, 3μm)色谱柱;以0.01 mol·L^(-1)磷酸二氢钾(磷酸调节pH值至3.0)∶乙腈(V/V,70∶30)为流动相,流速为1.0 mL·min^(-1),检测波长210 nm,柱温40℃。结果在上述色谱条件下,盐酸伊伐布雷定中的2个卤代烷烃类遗传毒性杂质(化合物Ⅰ和Ⅱ)分离效果良好;化合物Ⅰ和Ⅱ的检测限分别为0.001 6 mg·L^(-1)和0.010 1 mg·L^(-1)(分别相当于主成分的0.000 3%和0.002%,化合物Ⅰ和Ⅱ的限度之和为0.01%),它们的定量限分别为0.005 4 mg·L^(-1)1和0.020 2 mg·L^(-1);化合物Ⅰ在0.005 0~0.201 7 mg·L^(-1)内、化合物Ⅱ在0.019 9~0.199 1 mg·L^(-1)内的峰面积与浓度呈线性关系(r分别为0.999 8和0.999 9);化合物Ⅰ和Ⅱ的平均回收率分别为106.1%和101.4%,RSD%分别为3.4%和3.9%(n=9);重复性结果显示,化合物Ⅰ和Ⅱ的6份加样回收溶液测定结果的RSD分别为5.0%和<0.1%。空白溶剂和原料药基质对化合物Ⅰ和化合物Ⅱ的测定无干扰。结论上述结果表明:所建的基因毒杂质检测方法的灵敏度高,准确可靠、专属性好,适宜于盐酸伊伐布雷定原料中卤代烷烃类遗传毒性杂质的测定。

Objective A method was established for determination of haloalkane genotoxic impurities in ivabradine hydrochloride by RP-HPLC.Methods A CAPCELL PAK MGⅡ C18( 4.6×150 mm, 3 μm) column was used at 40 ℃,with 0.01 mol·L potassium dihydrogen phosphate aqueous solution(pH adjusted to 3.0 by phosphoric acid)-acetonitrile(70∶30,V/V)as the mobile phase at a flow rate of 1.0 mL·min^(-1).The detection wavelength is at 210 nm.Under the above chromatographic conditions.Results the resolution between the target genotoxic impurities(compounds Ⅰ and Ⅱ) was greater than 1.5.The detection limits of compounds I and II were 0.001 6 mg·L^(-1) and 0.010 1 mg·L^(-1),respectively, which corresponds to 0.000 3% and 0.002% of API(limits: sum of compounds I and II is less than 0.01%).Meanwhile, the limit of quantification compounds Ⅰ and Ⅱ were 0.005 4 mg·L^(-1)and 0.020 2 mg·L^(-1),respectively.Compound I in the range of 0.005 0-0.201 7 mg·L^(-1)the range of 0.019 9-0.199 1 mg·L^(-1) showed linear responses.And blank solvent has no interference on the determinations of compound Ⅰ and Ⅱ.Conclusion The above results showed the method is highly sensitive, accurate, and specific, which can be used for the determination of haloalkanes genotoxic impurities in ivabradine hydrochloride.