响应面法优化农产品中抗性淀粉的体外检测

Optimization of in vitro detection methods for resistant starch in agricultural products by response surface methodology

目的采用响应面法优化农产品中抗性淀粉(resistant starch,RS)的体外检测方法。方法选择谷类、薯类、豆类3类抗性淀粉主要来源的农产品,设计单因素试验和Box-Behnken试验。单因素试验考察了水浴时间、淀粉葡萄糖苷酶(amylglucosidase,AMG)作用温度、AMG作用时间、AMG添加量和离心力与离心时间5个条件,并选择AMG作用温度、AMG作用时间、AMG添加量来设计Box-Behnken试验。结果根据单因素试验,确定了水浴时间为14 h,离心条件选择4500 g离心5 min;根据响应面试验确定对RS含量检测结果影响程度排序为AMG添加量>AMG作用时间>AMG作用温度。结合单因素响应面试验,确定最佳检测方法为水浴14 h,4500 g离心5 min,AMG作用温度49℃、AMG添加量0.2 mL、AMG作用时间38 min。结论优化后的方法重复性良好,适用于玉米、马铃薯和红豆3类样品,且有望适用于其他主要农产品。

Objective To optimize the in vitro detection method of resistant starch(RS)in agricultural products by response surface methodology.Methods Three types of agricultural products,which were the main sources of resistant starch from cereals,tubers,and beans,were selected,and the single factor test and Box-Behnken test were designed.Five conditions:water bath time,amylglucosidase(AMG)action temperature,AMG action time,AMG addition amount,centrifugal force and centrifugal time were investigated by the single factor test,and AMG action temperature,AMG action time and AMG addition amount were selected to design the Box-Behnken test.Results According to the single factor test,the water bath time was determined as 14 h,and the centrifugation condition was 4500 g for 5 min;according to the response surface experiment,the order of influence on RS content detection results was AMG addition>AMG action time>AMG action temperature.Combined with the single factor response surface test,the best detection method was determined as water bath for 14 h,4500 g centrifugation for 5 min,AMG action temperature 49℃,AMG addition amount 0.2 mL and AMG action time 38 min.Conclusion The optimized method has good repeatability,which is suitable for corn,potato and red bean,and is expected to be suitable for other main agricultural products.