微生物发酵法生产去氢表雄酮检测方法的建立  

Establishment of determination method for DHEA production by microbial fermentation

作　　者：张磊[1] 靳魁奇[1] 刘宇鹏[1] 

机构地区：[1]河南大学生命科学学院生物工程研究所

出　　处：《食品与发酵工业》2015年第1期216-219,共4页Food and Fermentation Industries

基　　金：河南省科技厅攻关项目(No.132102210387);河南大学研究生优秀学位论文培育计划(Y1425012)

摘　　要：建立了一个微生物发酵生产去氢表雄酮（DHEA）的检测方法,采用气相色谱法同时测定发酵液中雄甾-4-烯-3,17-二酮（4-AD）、去氢表雄酮甲氧基甲基醚和DHEA含量。发酵液经乙酸乙酯萃取,可直接进样分析。气相色谱条件为：FID检测器,DB-1型（30 m×0.53 mm×0.25 m）毛细管柱;柱温280℃,进样口280℃,检测器310℃;载气为高纯N2,流速1 m L/min。在实验室条件下,发酵液中各主要成分分离情况良好。结果表明：4-AD、去氢表雄酮甲氧基甲基醚和DHEA的浓度线性范围分别为：4-AD为0.2~1.0 g/L（R2=0.996）;去氢表雄酮甲氧基甲基醚为0.1~0.9 g/L（R2=0.992）;DHEA为0.2~1.0 g/L（R2=0.998）,平均回收率依次为（97.88±0.74）%,（99.39±1.07）%和（98.91±1.09）%。该方法具有步骤简单,重现性好,准确度和灵敏度高的特点。A gas chromatography （GC） method for the simultaneous determination of Androst-4-ene-3, 17-dione （4-AD） , 3β-methoxymethyl-ether-dehydrepiandrosterone and dehydroepiandrosterone （DHEA） from a culture of mi- crobial fermentation for the production of transformable 3β-methoxymethyl-ether-phytosterols was described. The sam- ples were quantitated by internal standard method, and they were separated by GC with DB-1 capillary column and detected by FID. The column and the injector were operated at 280 ℃, the detector was set as 310 ℃ and the carrier gas was nitrogen, flowing at a rate of 1 mL min-1 The samples were quantitated by internal standard method. Under the conditions, all components in culture were well separated and rectilinear responses were obtained in the range of 0.2 ~ 1.0 g/L （4-AD）, 0. 1 - 0.9 g/L （ 3β-methoxymethyl-ether-dehydrepiandrosterone）, and 0.2 - 1.0 g/L （DHEA）. The correlation coefficients （R2） are 0. 996, 0. 992 and 0. 996 respectively. Recoveries of 4-AD, 3β-me- thoxymethyl-ether-dehydrepiandrosterone and DHEA added to cuhure were （97.88 ± 0.74） % , （99.39 ± 1.07） % and （98.91 ± 1.09）% respectively. The method is accurate, rapid and suitable for routine analvsis.

关 键 词：气相色谱 雄烯-4-烯-3 17-二酮 去氢表雄酮甲氧基甲基醚 去氢表雄酮 检测 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]