摘要
目的建立可同时检测绿袍散及其原料中5种(金胺O、柠檬黄、日落黄、金橙Ⅱ和酸性橙10)非法添加黄色素的高效液相色谱二极管阵列检测器(ultra performance liquid chromatography-diode array detector,UPLC-DAD)检测方法,用于快速筛查绿袍散及原料中的非法添加染色色素。方法色谱柱为Acquity BEH C 18色谱柱(2.1 mm×100 mm,1.7μm);流动相分别以甲醇为流动相A、0.02 mol/L乙酸铵为流动相B进行梯度洗脱;柱温40℃;流速0.3 ml/min,检测波长:金胺O和柠檬黄为432 nm,日落黄、金橙Ⅱ和酸性橙10为484 nm;进样量:2μl。采用超高效液相色谱串联质谱(ultra performance liquid chromatography tandem mass spectrometry,UPLC-MS)对UPLC-DAD检出色素的疑似阳性样品进行佐证和确认。结果5种色素对照品溶液在一定浓度范围内线性关系良好,线性相关系数(r)均大于0.9997;精密度良好,相对标准偏差(RSD)均小于1.03%;重复性良好;柠檬黄、日落黄、酸性橙10、金胺O和金橙Ⅱ峰面积的RSD分别为2.73%,2.92%,1.97%,1.56%和1.58%,表明该供试品溶液在24 h之内稳定性良好;回收率均在96.33%-99.34%之间。10批绿袍散样品中均未检出上述5种非法添加黄色素,6批原料药中仅有一批原料药(黄柏)中检出金胺O。结论UPLC-DAD技术应用快速、灵敏,且容易推广,可作为筛查绿袍散及其原料中5种非法添加黄色染料的检查方法。
Objective To establish an ultra performance liquid chromatography-diode array detector method for quick and simultaneous determination of five yellow pigments illegally added into Lvpao powder and its active pharmaceutical ingredients.Methods The Acquity BEH-C 18(2.1 mm×100 mm,1.7μm)chromatography column was performed for separation with methanol(A)and 0.02 mol/L ammonium acetate(B)as the mobile phase by gradient elution,the column temperature was maintained at 40℃with flow rate of 0.3 ml/min,the detection wavelength was 432 nm for auramine O and lemon yellow,and 484 nm for sunset yellow,golden orangeⅡand acid orange 10,and the injection volume was 2μl.UPLC-MS was used to corroborate and confirm the suspected positive samples of pigments detected by UPLC-DAD.Results The five kinds of pigment reference solution showed a good linear relationship within a certain concentration range,and the linear correlation coefficient was greater than 0.9997.The precision was good,the relative standard deviation(RSD)was less than 1.03%,and the repeatability was good.The RSDs of the peak areas of acid orange 10,auamine O and golden orangeⅡwere 2.73%,2.92%,1.97%,1.56%and 1.58%,respectively,indicating that the test solution was stable within 24 h.The recovery rate was 96.33%-99.34%.None of the above five illegally added yellow pigments were detected in the 10 batches of Lvpao powder samples,and only one batch of raw materials(Cork)was detected in the six batches of raw materials.Conclusion UPLC-DAD technology is fast,sensitive,and easy to popularize,and can be used as an inspection method for screening Lvpao powder and five illegally added yellow dyes in its raw materials.
作者
莫显超
朱海媖
马婵娟
MO Xianchao;ZHU Haiying;MA Chanjuan(Department of Stomatology,Guangzhou Red Cross Hospital,Guangzhou 510220,China;Department of pharmacy,Guangzhou Red Cross Hospital)
出处
《山西医科大学学报》
CAS
2021年第10期1363-1368,共6页
Journal of Shanxi Medical University
关键词
绿袍散
原料药
非法添加
金胺O
柠檬黄
日落黄
金橙Ⅱ
酸性橙10
含量测定
液质联用
Lvpao powder
active pharmaceutical ingredients
illegal additive
golden auramine O
lemon yellow
sunset yellow
orangeⅡ
acid orange 10
content determination
UPLC-MS
