纳豆芽孢杆菌发酵液中嘌呤碱基的检测

Determination of purine base in bacillus natto fermentation broth

摘要: 采用反相离子对色谱法对纳豆芽孢杆菌发酵液中的嘌呤碱基进行检测,优化后的检测条件为:色谱柱,Hitachi La Chrom C18(4.6mm×250mm,5μm);紫外检测波长254nm;流动相:甲醇∶四丁基氢氧化氨∶乙酸∶水=10∶1.485∶1.485∶987.03(v/v/v/v);流速:0.7mL/min;柱温:25℃。检测结果表明该方法标准曲线良好:A、G、H、X四种嘌呤碱基的线性范围分别为0.5~20.0、1.0~50.0、1.0~50.0、0.5~20.0mg/L,相关系数均大于0.9991,方法回收率为96.9%~102.4%。基于优化后的检测方法,检测得出纳豆发酵前后发酵液中总嘌呤含量分别为41.33、84.99mg/L,说明嘌呤碱基含量在发酵过程中有所增加。

Abstract: Purine bases were detected by reverse phase ion pair chromatograph. The determination condition could be concluded as follows:Column was Hitachi La Chrom C18 (4.6mm×250mm, 5μm) . UV detector wave was 254nm, mobile phase was 0.7mL/min, column temperature was 25℃. Detection results demonstrated that the standard curve was well, the linearity range of A, G, H, X were 0.5²0.0, 1.0⁵0.0, 1.0⁵0.0, 0.5²0.0mg/L respectively, the correlation coefficient was above 0.9991 and recovery scope was 96.9%¹02.4%. Based on optimized detection method, purine bases in both before and after fermentation was 41.33mg/L and 84.99mg/L, which demonstrateed that purine bases were increasing during fermentation.