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

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( R²) 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.