高效液相色谱法测定酵母粉腺嘌呤含量

目的建立高效液相色谱法测定酵母粉腺嘌呤的含量,为酵母粉质量评价奠定方法学基础。方法将酵母粉经过破壁处理后用高氯酸消解制备提取液,通过条件实验确定高效液相色谱法分离条件并测定腺嘌呤含量。结果采用Diamonsil C18柱,以0.02 mol/L磷酸盐缓冲液(pH 6.0)为流动相,流速为0.8 mL/min,可以将腺嘌呤与其它成分有效分离。采用121℃高温高压-SDS法进行酵母粉前处理能有效破壁和促进核酸物质的释放,腺嘌呤测定值更接近其实际含量。结论采用高温高压-SDS法制备酵母粉提取液、高效液相色谱测定腺嘌呤含量的方法简单易行、重现性好,测定结果可作为酵母粉质量评价的重要定量指标,对发酵生产有重要指导意义。

Determination of adenine content in yeast powder by HPLC

Purpose To establish a method for the determination of adenine content in yeast powder by HPLC and to lay a methodology basis for the quality assessment of yeast powder.Methods After the yeast powder was pre-treated by way of wall-breaking,degraded with perchloric acid,the extracts were prepared for the sepration and determination of adenine by HPLC.Results Under the following conditions:Diamonsil C18 column,pH 6.0,0.02 mol/L phosphate buffer as mobile phase and flow rate 0.8 mL/min,adenine could be separated effectively from other components.The yeast powder pre-treatment experiments showed that yeast cell wall could be effectively broken through 121 ℃ autoclaving-SDS pre-treatment and nucleic acid was promoted to release,and the determination results of adenine were closer to the natural value.Conclusion The method using autoclaving-SDS to prepare yeast powder extracts and determining the adenine content by HPLC,is simple and reproducible.Analysing results would be an important evaluation index to investigate yeast powder quantitatively,which is of significance for guiding the production of inosine fermentation.