HPLC波长程序法同时检测非发酵豆制品中碱性橙和碱性嫩黄O的含量

Simultaneous detection of Basic Oranges dyes and Basic Flavine O dye contents in non- fermented soybean products by HPLC wavelength program method

目的:建立非发酵豆制品中的碱性橙2、碱性橙21、碱性橙22和碱性嫩黄O的HPLC波长程序法同时检测的方法。方法:样品采用不同溶剂超声提取,提取液离心,上清液旋转蒸发浓缩。样液以甲醇-乙酸铵溶液为流动相,流速1.0 m L/min,经C18反相色谱柱分离,设定波长程序实现四种染料的同时检测,并进行了方法学评价。结果:选择乙醇为超声提取的溶剂。碱性橙2、碱性橙21和碱性橙22和碱性嫩黄O的线性范围为0.0~10.0μg/m L,相关系数R^2≥0.9990。检出限在0.01~0.05 mg/kg之间,定量限在0.046~0.098 mg/kg之间。三个浓度(5.00、10.00、20.00 mg/kg)的加标回收率的范围为90.38%~102.16%,相对标准偏差(RSD)范围为1.37%~6.93%。结论:四种碱性染料的HPLC波长程序检测方法满足方法评价的要求,具有灵敏、准确、快速的特点。

Objective： To establish a method for simultaneous detection of Basic Orange 2,Basic Orange 21,Basic Orange 22 and Basic Flavine O in non- fermented soybean products using HPLC wavelength program method.Methods： Samples solutions were extracted with different solvents under ultrasonic condition. The extract was centrifuged,and the upper solution was concentrated by rotary evaporator. The analysis was carried on C18 column.The mobile phase was aqueous solution of methanol- ammonium acetate at the flow- rate of 1.0 m L / min,programmed wavelength ultraviolet detection was performed to simultaneously determine four dyes.In addition,the detection methodologies were evaluated. Results： Ethanol was selected as ultrasonic extraction solvent. The linear range of this method was 0.0~10.0 μg / m L for Basic Orange 2,Basic Orange 21,Basic Orange 22 and Basic Flavine O,and the correlation coefficient（ R^2） was higher than 0.9990.The limit of detection was between 0.01 mg /kg to0.05 mg / kg,the limit of quantitation was between 0.046 to 0.098 mg / kg. The recoveries range of sample added standards of three concentrations（ 5.00,10.00,20.00 mg / kg） were 90.38% ~ 102.16%,RSD was 1.37% ~6.93%.Conclusion： The HPLC wavelength program detection method of Basic Orange and Basic Flavine O is able to meet requirements of the methodological evaluation,and it is sensitive,accurate and rapid.