HPLC-HG-AFS法测定活血止痛胶囊中不同汞形态含量的方法研究

Speciation analysis of mercury in Huoxue Zhitong capsules by HPLC-HG-AFS

目的:探讨活血止痛胶囊中二价汞、甲基汞、乙基汞三种汞形态的含量测定方法,同时为活血止痛胶囊风险防控提供数据支持。方法:采用高效液相色谱-氢化物发生-原子荧光光谱法(HPLC-HG-AFS)测定不同批次活血止痛胶囊中3种汞形态的含量,色谱条件:C 18反相色谱分离柱(250 mm×4.6 mm,5μm),以乙腈-0.06 mol·L^(-1)乙酸铵溶液(含0.01 mol·L^(-1)L-半胱氨酸)(5∶95)为流动相;氢化物发生参数:氧化剂为1%过硫酸钾-0.35%氢氧化钾的混合溶液;还原剂为2%硼氢化钾-0.35%氢氧化钾的混合溶液,载流为7%盐酸溶液,负高压285 V,原子化器高度为10 mm。结果:三种汞形态在1.00~10.0μg·L^(-1)范围内线性关系良好,相关系数均大于0.999,检测限在0.088~0.131 ng·L^(-1)之间,平均回收率在91.98%~93.10%之间,RSD均小于5.73%(n=3)。采用HPLC-HG-AFS法测定了不同批次活血止痛胶囊中三种汞形态的含量,结果表明均低于检测限。结论:本法经方法学验证,可用于活血止痛胶囊等中成药中汞形态含量测定,为中成药的安全性评价提供参考。

Objective:To determine divalent mercury,methylmercury and ethylmercury in Huoxue Zhitong capsules,and to provide data support for the risk assessment and management of traditional medicines products.Methods:High performance liquid chromatography-hydrogenation-atomic fluorescence spectrometry(HPLC-HG-AFS)was used to determine the content of three forms of mercury(divalent mercury,Hg^(2+);methyl mercury,CH 3 Hg;ethyl mercury,C_(2)H_(5)Hg)in different batches of Huoxue Zhitong capsules.Chromatographic conditions were C 18 column(250 mm×4.6 mm,5μm)and acetonitrile-0.06 mol·L^(-1)ammonium acetate solution(containing 0.01 mol·L^(-1)L-cysteine)(5∶95)as a mobile phase.Hydride generation parameters were oxidant agent of 1%potassium persulfate-0.35%potassium hydroxide mixed solution,reducing agent of 2.0%potassium borohydride-0.35%potassium hydroxide mixed solution,carrier solution of 7%hydrochloric acid,negative high pressure of 285 V,and atomizer height of 10 mm.Results:The linear range of mercury species was 1.00-10.0μg·L^(-1)with r>0.999.The detection limit values were 0.088-0.131 ng·L^(-1),and the average recoveries ranged from 91.98%to 93.10%with RSD of<5.73%(n=3).Three mercury forms in different batches of Huoxue Zhitong capsules by HPLC-HG-AFS were below the detection limits.Conclusion:The validated method can be used for the determination of mercury speciation in Huoxue Zhitong capsules and other Chinese patent medicines,which provides a reference for the safety assessment of Chinese patent medicines.