聚酰胺固相萃取-HPLC法同时测定中药材中16种合成酸性色素

Simultaneous determination of sixteen synthetic acid pigments in medicinal Chinese herbs by polyamide SPE-HPLC

目的针对中药材及饮片染色现象,建立同时测定金橙Ⅰ、亮蓝G、亮蓝、柠檬黄、日落黄、亮黄、金橙Ⅱ、酸性橙10、赤藓红、酸性红73、苋菜红、酸性红18、诱惑红、曙红钠、羊毛绿、酸性黑1等16种合成酸性色素的HPLC检测方法。方法样品用甲醇-0.1%甲酸溶液提取后,再经聚酰胺固相萃取去除大部分基质干扰,采用Agilent ZORBAX C18色谱柱（4.6 mm×250 mm,5μm）,以甲醇-20 mmol/L乙酸铵为流动相,梯度洗脱,以二极管阵列检测器测定。结果在丹参、山茱萸、槐米、黄连四种基质中,这16种色素在50 ng/m L至50μg/m L范围内线性关系良好。其中前14种色素可定量,定量检测限为1.13~8.77 mg/kg,回收率为61.4%~112.0%,相对标准偏差为1.2%~5.4%。结论本方法适用范围广,已应用于实际样品测定。

AIM To develop an HPLC method for simultaneously determining sixteen synthetic acid pigments,including orangeⅠ,brilliant blue G,brilliant blue,tartrazine,sunset yellow,brilliant yellow,orange Ⅱ,orange G,erythrosin B,acid red 73,amaranth,acid red 18,allura red,eosin,green S,and acid black 1,to distinguish the dyeing from medicinal Chinese herbs. METHODS The samples were extracted by methanol-0. 1% formic acid,then purified by polyamide solid phase extraction（SPE）. A reversed phase column,Agilent ZORBAX C18（4. 6 mm × 250 mm,5 μm） was used with methanol-20 mmol/L ammonium acetate as the mobile phase in a gradient elution mode,and the diode array detector was used for the determination. RESULTS Sixteen pigments presented in Danshen（Salvia miltiorrhizae Radix et Rhizoma）,Shanzhuyu（Corni Fructus）,Huaimi（Sophorae Flos） and Huanglian（Coptidis Rhizoma） possessed good linear correlation in the concentration range from 50 ng/m L to 50 μg/m L. The first fourteen pigments could be quantified,their quantitative detection limits ranged from 1. 13 mg/kg to 8. 77 mg/kg,and their recoveries ranged from 61. 4% to 112. 0%,the RSDs ranged from1. 2% to 5. 4%. CONCLUSION This method is universal in the determination of synthetic pigments in practical application.