HPLC-MS/MS法测定盐酸拉贝洛尔中潜在基因毒性杂质

Determination of potential genotoxic impurities in labetalol hydrochloride by HPLC-MS/MS method

为了更好地控制盐酸拉贝洛尔中潜在的基因毒性杂质,保证用药安全,建立了高效液相色谱-质谱(HPLC-MS/MS)法检测盐酸拉贝洛尔中潜在遗传毒性杂质N-亚硝基拉贝洛尔。液相色谱条件:以Waters ACQUITY UPLC CSH C_(18)(150 mm×3.0 mm,1.7μm)为色谱柱,流动相A为0.01 mol/L甲酸铵-0.1%甲酸水溶液,流动相B为乙腈溶液,梯度洗脱,流速为0.5 mL/min,柱温为50℃,进样体积为10μL。以电喷雾电离源为离子源,负离子模式,采用多反应监测,建立质谱条件,对盐酸拉贝洛尔中N-亚硝基拉贝洛尔进行定量检测。结果表明:N-亚硝基拉贝洛尔质量浓度在1.01~100.60 ng/mL范围内有良好的线性关系;低、中、高3个浓度的加样回收率(n=3)为96.84%~99.53%;检测限和定量限分别为0.01 ng/mL和0.03 ng/mL;6批盐酸拉贝洛尔样品中N-亚硝基拉贝洛尔的含量为0.37~0.79 ng/mL。所提出的方法灵敏度高、专属性强,可用于测定盐酸拉贝洛尔中的N-亚硝基拉贝洛尔,为盐酸拉贝洛尔的质量控制提供参考。

In order to better control the potential genotoxic impurities in labetalol hydrochloride and ensure the safety of medication,a high-performance liquid chromatography-mass spectrometry(HPLC-MS/MS)method was established to detect potential genotoxic impurities N-nitroso-labetalol in labetalol hydrochloride.The liquid chromatographic conditions were as following:the chromatographic column was Waters ACQUITY UPLC CSH C_(18)(150 mm×3.0 mm,1.7μm),mobile phase A was 0.01 mol/L ammonium formate-0.1%formic acid aqueous solution,mobile phase B was acetonitrile solution,the gradient elute was adopted,flow rate was 0.5 mL/min,the column temperature was 50℃,and the injection volume was 10μL.Mass spectrometry conditions were established by using the electrospray ionization source as the ion source,negative ion mode and the multiple reaction monitoring mode,so that N-nitroso-labetalol in labetalol hydrochloride was quantified.The results indicate that the calibration curves is in good linearity in the range of 1.01~100.60 ng/mL;The recoveries(n=3)at low,middle and high spiked concentrations are within 96.84%~99.53%;The limit of detection is 0.01 ng/mL,and the limit of quantification is 0.03 ng/mL;The amount of N-nitroso-laelalol in six batches of the hydrochloride samples is 0.37~0.79 ng/mL.The proposed method is sensitive and accurate,and can be used for the quantification of N-nitroso-labetalol in labetalol Hydrochloride,providing some reference for the quality control of labetalol hydrochloride.