| 摘要: |
| 在用RP-HPLC法C18色谱柱和紫外检测器检测发酵液中丙氨酸时，由于发酵液中存在丙酮酸，而丙酮酸对紫外吸收过强，且两者峰之间有重叠，导致无法准确检测出丙氨酸含量。为解决此问题，本文在RP-HPLC检测前首先对丙氨酸发酵液进行了乙醇沉淀预处理，实现了丙酮酸和丙氨酸在检测进样前的分离，消除了丙酮酸信号对丙氨酸的影响，能准确测定出丙氨酸含量。达到有效分离的发酵液与无水乙醇最佳比例为1∶14；经氨基酸自动分析仪检测发酵液中丙氨酸实际含量为92.60g/L；经RP-HPLC检测，沉淀中丙氨酸的含量为为92.04 g/L,丙氨酸的回收率高达99.40%。本研究利用乙醇对丙氨酸发酵液进行预处理，使RP-HPLC法可以运用于检测发酵液中丙氨酸含量，适用于工厂发酵生产过程中丙氨酸的快速检测。 |
| 关键词: 丙氨酸 乙醇预处理 氨基酸自动分析 反相高效液相色谱分析 |
| DOI：10.14188/j.ajsh.2018.06.010 |
| 分类号:O657.7 |
| 基金项目:国家“十二五”支撑计划（2012BAD27B03），山东技术创新计划（201720311004） |
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PAN Hailiang1,LI Guiling2,WANG Can1,CUI Guangshui2,WANG Jinhua1
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1.Key Laboratory of Fermentation Engineering of Ministry of Education, Hubei University of Technology, Wuhan 430068, Hubei, China;2.Kai Xiang biochemical Co., Ltd, Rizhao 262300, Shandong,China
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| Abstract: |
| In fermentation broth, alanine could not be accurately detected by RP-HPLC on C18 column and UV detector, because of the existence of pyruvic acid which has a strong UV absorption, and the overlap between two peaks. In order to solve this problem, ethanol precipitation pretreatment of alanine fermentation broth was carried out before RP-HPLC detection. The separation of pyruvic acid and alanine was realized before the detection of alanine. The interference of pyruvic acid signal on alanine was eliminated and the content of alanine was accurately determined. The optimum ratio of fermentation broth to anhydrous ethanol was 1∶14 for effective separation；the actual content of alanine in fermentation broth was 92.60 g/L by automatic amino acid analyzer； the content of alanine in precipitation was 92.04 g/L by RP-HPLC, and the recovery rate of alanine was 99.40%. In this study, ethanol was used to pretreat alanine fermentation broth, so that RP-HPLC method can be used to detect alanine content in fermentation broth, which is suitable for rapid detection of alanine in factory fermentation process. |
| Key words: alanine ethanol pretreatment amino acid automatic analyzer reversed-phase high-performance liquid chromatography(RP-HPLC) |
