气相色谱法测定植物甾醇发酵液中雄甾烯酮和植物甾醇的含量

Determination of 4-AD,ADD and Phytosterols in Culture of Transformable Phytosterol by Gas Chromatographic

采用气相色谱仪建立了一个同时测定植物甾醇发酵液中雄甾-4-烯-3，17-二酮（Androst-4-ene-3，17-dione，4-AD）、雄甾-1，4-二烯-3，17-二酮（Androst-1，4-diene-3，17-dione，ADD）和植物甾醇含量的外标分析方法．发酵液经离心后，二氯甲烷提取，可直接进样分析．气相色谱条件为：FID检测器，SE-54型（15m×0．25mm×0．25gm）毛细管柱；柱温260℃，进样口和检测器温度均为300℃；载气为高纯N2，流速1mL／min．在实验条件下，发酵液中各主要成分和杂质分离良好．结果表明：4-AD、ADD和植物甾醇各组分的浓度与Ai／As的线性范围4-AD为0．5-4g／L（R=0．9999）；ADD为0．5～4g／L（R=0．9992）；菜籽甾醇为0．09～0．86g／L（R=0．9991）；菜油甾醇为0．14～1．39g／L（R=0．9995）；豆甾醇为0．03～0．27g／L（R=1）；谷甾醇为0．24～2．4g／L（R=0．9991），平均回收率4-AD、ADD和上述植物甾醇依次为（96．37±0．77）％，（95．64±1．01）％，（99．6±278）％，（9866±2．19）％，（96．85±2．19）％，（97．52±2．9）％．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 ）, Androst-1,4-diene-3, 17-dione （ ADD ）and phytosterols from aculture of transformable phytosterols is described.The samples were quantitated by external standard method, and they were separated by GC with SE-54 capillary column and detected by FID.The column was operated at 260 ℃, with the injector at 300 ℃ and the carrier gas was nitrogen, flowing at a rate of 1 mL/min.Under the conditions, all contents of culture are well separated and rectilinear responses are obtained in the range of 0.09～0.86 g/L （stigmasterol）, 0.14～1.39 g/L （campesterol）, 0.03～0.27 g/L （brassicasterol）, 0.24～2.4 g/L （ 13-sitosterol ）, 0.5～4 g/L （ 4-AD ）and 0.5～4 g/L （ ADD ）.The correlation coefficients （ R ）are 0.9991,0.9995,1,0.9991, 0.9999 and 0.9992 respectively.Recoveries of 4-AD, ADD and phytosterols added to culture are （96.37±0.77）%, （ 95.64±1.01 ）%, （ 99.6±2.78 ）%, （ 98.66±2.19 ）%, （ 96.85±2.19 ）%, （ 97.52±2.9 ）%.The method is accurate rapid and suitable for routine analysis.