摘要
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。
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 condi- tions：Diamonsil Cls 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 im- portant evaluation index to investigate yeast powder quantitatively, which is of significance for guiding the production of inosine fermentation.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2012年第5期552-554,558,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
国家科技部科技人员服务企业行动项目(2009GJD00044)
河南省教育厅自然科学基金项目(2011B180032)
