气相色谱法测定酵母转化2,5-己二酮发酵液中的主要成分

Determination of targets in fermentation of transformation of 2,5-hexanedione with yeast cells by GC

采用毛细管柱INNOWAX(30m×0.25mm×0.25μm),以1,3-丙二醇为内标物对生物转化发酵液中底物2,5-己二酮、产物2,5-己二醇及中间产物5-羟基-2-己酮进行气相色谱分析。优化分析条件为:进样口为SPL,温度250℃;FID检测器,温度250℃;柱温140℃;载气为氮气,线速度3.0cm/min;分流进样,进样量1.0μL,分流比40∶1。在10min内,3个组分能得到完全分离,回收率分别为98.8%、103.0%和99.3%,相对标准偏差分别为0.92%、2.9%和3.9%。

In this study, a capillary column INNOWAX （30 m× 0.25 mm i.d., 0.25 μm） was selected to measure simultaneously substrate 2,5-hexanedione, product 2,5-hexanediol and intermediate 5-hydroxy-2-hexanone in the fermentation of biotransformation, with 1,3-propylene glycol as an inner standard by gas chromatography. The optimal temperatures of the SPL sample receiver, the FID detector, and the column were determined to be 250 ℃, 250 ℃ and 140 ℃, respectively, with nitrogen as the cartier at the speed of 3.0 cm/min. The injection volume of the sample was set at 1.0μL with the shunting ratio of 40：1. The results showed that three targets can be completely separated within 10 min. The recoveries of 2, 5-hexanedione, 2, 5-hexanediol and 5-hydroxy-2-hexanonewere reached as high as 98.8%, 103.0% and 99.3%, with the relative standard deviations （RSD） were 0.92%, 2.9% and 3.9%, respectively. This method was suitable for the determination of three targets in the fermentation of transformation of 2,5- hexanedione by baker＇s yeast, with high reproducibility and good linearity.