柱前衍生气相色谱法分离手性二醇类化合物

Pre-column Derivatization GC Method of Chiral Diols

本文建立了柱前衍生气相色谱法分离1,3-丁二醇、2,3-丁二醇、2,5-己二醇和3,6-辛二醇4种二醇类化合物的手性分离方法。分别使用N-甲基-N-(三甲基硅烷)三氟乙酰胺和三氟乙酸酐为衍生剂与二醇化合物反应,生成相应的衍生物,用CP-Chirasil-DexCB色谱柱分离二醇对映体,采用分流进样方式,检测器为氢火焰离子化检测器(FID)。在选定的色谱条件下,四种手性化合物分别在2.50~8×10^(3)μg/mL、1.30~8×10^(3)μg/mL、3.5~1×10^(4)μg/mL、5~1×10^(4)μg/mL范围内呈线性关系,相关系数分别为0.9994、0.9997、0.9990和0.9971,手性分离度均达到2以上,检测限分别为0.8μg/mL、0.4μg/mL、1.8μg/mL、2.5μg/mL。该方法适用于生物酶催化反应所生成的(2R,3R)-2,3-丁二醇、(R)-1,3-丁二醇、(2S,5S)-2,5-丁二醇和(3S,6S)-3,6-辛二醇的检测以及酶促反应过程的监控。

A method of the chiral separation of 1,3-butanediol,2,5-hexadiol,3,6-octanediol and 3,6-octanediol enantiomers via derivatization-capillary column gas chromatography was established and validated.The diol enantiomers were derivatized with N-methyl-n-(trimethylsilane)trifluoroacetamide or trifluoroacetic anhydride and then were separated by CP-Chirasil-Dex CB column,split sampling,and FID as detector.The linearity of the methods were proven well with the correlation coefficients were 0.9994,0.9997,0.9990,and 0.9971,as well as the detection ranges of 2.50-8×10^(3)μg/mL,1.30-8×10^(3)μg/mL,3.5-1×10^(4)μg/mL and 5-1×10^(4)μg/mL.All the resolution factors of enantiomers separation were above 2.The detection limits of 4 analytes were 0.8μg/mL,0.4μg/mL,1.8μg/mL,and 2.5μg/mL,respectively.The method is applied in the determination of(R)-1,3-butanediol,(2R,3R)-2,3-butanediol,(2S,5S)-2,5-hexanediol and(3S,6S)-3,6-octanediol which were produced by enzymatic reaction and the enzymatic reaction monitoring as well.