杯碟法测定泛酸含量的应用研究

Application of Cylinder Plate Method for Determination of Pantothenic Acid

为建立一种快速高效便捷、高通量测定泛酸含量的方法,该研究结合了传统微生物法和杯碟法的优点,利用植物乳杆菌对泛酸的特异性,开发了杯碟法测定泛酸含量的方法。同时对试验菌株的制备方法和测定用平板的制备方法进行有效的质量控制。研究结果表明,该批次菌株在含0.5%琼脂的泛酸测定用平板中菌浓度OD_(600)为0.030时,生长圈清晰,呈较规则圆形。该方法在泛酸浓度为10~200 ng/mL范围内,标准曲线线性关系良好,R^(2)值可达到0.99以上。杯碟法与国标法所测结果都与标准样品的标示值相符,RSD均小于5%,无明显差异。重复性试验的RSD分别为4.5%、4.5%、3.7%、4.5%和3.8%。方法具有步骤便捷、检测快速、结果准确、重复性好和检测成本低等优势,能克服传统微生物法的缺陷与不足,为泛酸含量测定方法的改进提供参考。

In order to establish a fast,efficient,convenient and high flux method for the determination of pantothenic acid content,this study combined the advantages of traditional microbial method and cylinder plate method,and utilized the specificity of Lactobacillus plantarum for pantothenic acid to develop the cylinder plate method for the determination of pantothenic acid content.At the same time,effective quality control was performed on the preparation method of the experimental strain and the preparation method of the assay plate.The research results showed that when the concentration OD_(600)of the batch of strains added to the pantothenic acid assay plate containing 0.5%agar was 0.030,the growth circle was clear and regular circle.The method has a good linear relationship with the standard curve in the pantothenic acid concentration range of 10~200 ng/mL,and the correlation coefficient was greater than 0.99.The results of both methods are consistent with the labeled values of the standard samples.The RSD is lower than 5%,and there is no significant difference.RSD values of repeatability test were 4.5%,4.5%,3.7%,4.5%,and 3.8%.The method has the advantages of convenient steps,rapid detection,accurate results,good repeatability and low detection cost,can overcome the defects and deficiencies of traditional microbial methods,and provides reference for the improvement of pantothenic acid content determination methods.