高效液相色谱法测定豆芽中6-苄基腺嘌呤含量的方法优化

Optimization of high performance liquid chromatography method for determination of 6-benzylaminopurine content in bean sprouts

目的建立高效液相色谱法测定豆芽中6-苄基腺嘌呤(6-benzylaminopurine,6-BA)含量的方法,验证6-BA快速检测试纸条(胶体金法)的检测结果。方法以6-BA含量为指标,研究样品的浸泡超声时间,离心转速,旋蒸温度,流动相比例等测定条件对豆芽中6-BA测定结果的影响。以绿豆芽为试样,对比高效液相色谱法和6-BA快速检测试纸条(胶体金法)的检测结果。结果6-苄基腺嘌呤在0.04~1.0μg/mL的浓度范围内线性关系良好,定量限为0.02 mg/kg,回收率为92.0%~106.2%,相对标准偏差为1.06%~2.31%。实际样品的检测结果与快速检测试纸条(胶体金法)的结果与一致。结论优化后的高效液相色谱法结果准确,回收率较好,适合用于豆芽中6-BA的测定。也可用于6-BA快检设备盲样的定量和稳定性验证,为快速有效测定豆芽中6-BA含量提供理论支撑。

Objective To establish a method for the determination of 6-benzylaminopurine(6-BA)in bean sprouts by high performance liquid chromatography,and to verify the test results of the 6-BA rapid test strip(colloidal gold method).Methods Using the 6-BA content as an index,the effects of the immersion ultrasonic time,centrifugation speed,spin temperature,and flow ratio on the measurement results of 6-BA in bean sprouts were studied.Mung bean sprouts were used as samples,and the results of high performance liquid chromatography and 6-BA rapid detection test strips(colloidal gold method)were compared.Results The 6-benzyl adenine had a good linear relationship in the concentration range of 0.04-1.0μg/mL.The limit of quantification was 0.02 mg/kg,the recoveries were 92.0%-106.2%,and the relative standard deviations were 1.06%-2.31%.The test results of the actual samples were consistent with those of the rapid test strip(colloidal gold method).Conclusion The optimized high performance liquid chromatography method has accurate results and good recovery rate,which is suitable for the determination of 6-BA in bean sprouts.It can also be used for the quantitative and stability verification of blind samples of 6-BA fast detection equipment,providing theoretical support for the rapid and effective determination of 6-BA content in bean sprouts.