微孔板式微生物法测定婴幼儿配方乳粉中叶酸、泛酸和生物素含量

Determination of folic acid, pantothenic acid, and biotin content in infant formula milk powder by microplate microbial method

中文关键词:  微孔板  微生物法  婴幼儿配方乳粉  叶酸  泛酸  生物素

英文关键词:microplate  microbial method  infant formula milk powder  folic acid  pantothenic acid  biotin

中文摘要:

      目的 建立微孔板式微生物法测定婴幼儿配方乳粉中的叶酸、泛酸和生物素含量的方法。方法 按照国标方法分别提取婴幼儿配方乳粉中的叶酸、泛酸和生物素，在96孔微孔板中接种含有相应测试菌悬液的培养基，再分别加入标准系列溶液和试样提取液，培养后用酶标仪测定吸光度，根据标准曲线分别计算试样中的叶酸、泛酸和生物素含量，并考察测定方法的检出限、精密度和加标回收率。结果 叶酸、泛酸和生物素标准曲线的相关系数均大于0.999，检出限分别为0.10 μg/100 g、0.013 mg/100 g、0.57 μg/100 g，相对标准偏差(relative standard deviations, RSDs)分别为1.02%、0.94%、1.08%，加标回收率分别为94.6%~100.0%、95.3%~103.3%、94.5%~100.1%。结论 该微孔板方法操作简单、成本较低，结果准确性和重现性良好，适用于婴幼儿配方乳粉中叶酸、泛酸和生物素含量的大批量检测。

英文摘要:

      Objective To establish a microplate microbial method for the determination of the content of folic acid, pantothenic acid, and biotin in infant formula milk powder. Methods The folic acid, pantothenic acid, and biotin were extracted from infant formula milk powder according to the national standard methods. The culture mediums containing the corresponding test bacterial suspension were inoculated into a 96-well microplate, and then the standard series solutions and sample extraction solutions were added separately. The absorbance value was measured using a microplate reader after cultivation. The content of folic acid, pantothenic acid, and biotin in the sample were calculated according to the standard curve, and the detection limits, quantification limits, precisions, and spiked recovery rates of the methods were investigated. Results The correlation coefficients of the standard curves for folic acid, pantothenic acid, and biotin were all greater than 0.999. The limits of detection were 0.10 μg/100 g, 0.013 mg/100 g, and 0.57 μg/100 g. The relative standard deviations (RSDs) were 1.02%, 0.94%, and 1.08%. The spiked recovery rates were 94.6%-100.0%, 95.3%-103.3%, and 94.5%-100.1%. Conclusion This microplate method is simple, cost-effective, and has good accuracy and reproducibility of results. It is suitable for large-scale detection of folic acid, pantothenic acid, and biotin content in infant formula milk powder.