响应曲面法优化HS-SPME-GC-MS测定天然牧草中挥发性成分工艺研究

Optimization of HS-SPME-GC-MS for determination of volatile components in natural forage by response surface methodology

为了优化顶空固相微萃取(HS-SPME)和气相色谱-质谱(GC-MS)联用测定天然牧草中挥发性成分的萃取条件,试验以色谱峰总峰面积为考察指标,在单因素试验的基础上,运用Box-Behnken响应曲面法设计萃取温度、萃取时间、解析时间和样品量4因素3水平的试验模型,建立回归方程,优化筛选萃取条件。结果表明:模拟得到的最佳萃取条件为萃取温度69.60℃、萃取时间58.39 min、解析时间2.83 min、样品量2.49 g,预测色谱峰总峰面积为9.958×10^9。为了提高萃取条件的可行性,将萃取条件修正为萃取温度70℃、萃取时间60 min、解析时间3 min、样品量2.5 g,验证色谱峰总峰面积为9.842×10^9,与模型预测值较接近。说明响应曲面法优化HS-SPME-GC-MS测定天然牧草中挥发性成分的萃取条件效果好,预测值可靠。

In ordertooptimize the extraction conditions of head space solid phase microextraction(HS-SPME)and gas chromatography-mass spectrometry(GC-MS)for the determination of volatile components in natural forage,the total peak area of the chromatographic peaks was usedas the index of investigation.a three-level model with four factors including extraction temperature,extraction time,resolution time and sample sizewas designed by usingBox-Behnken response surface methodOn the basis of single factor test,and the regression equation was established and extraction conditions were optimized.The results showed that the optimal extraction temperature was 69.60°C,the optimum extraction timewas 58.39 min,the optimumanalysis time was2.83 minand the optimumsample volume was 2.49 g.The total peak area of the predicted peak was 9.958×10^9.In order to improve the feasibility of the extraction conditions,the extraction conditions were corrected as follows:extraction temperaturewas 70°C,extraction time was 60 min,analysis time was 3 min,sample amount was 2.5 g,and the total peak area of the chromatographic peak was 9.842×10^9,which was close to the predicted value of the model.The results indicated that the HS-SPME-GC-MS method optimized byresponse surface methodologywas effective and reliable in determining thevolatile constituents in natural forage.