基于固相微萃取箭型-气相色谱-质谱法测定铜绿微囊藻中挥发性代谢物  

Determination of Volatile Metabolites in Microcystis Aeruginosa Using Headspace-Solid Phase Microextraction Arrow Combined with Gas Chromatography-Mass Spectrometry

作　　者：虞锐鹏 王利平[2] 赵晨凯[1] 吴胜芳 宋启军[1] YU Rui-Peng;WANG Li-Ping;ZHAO Chen-Kai;WU Sheng-Fang;SONG Qi-Jun(School of Chemical and Material Engineering,Wuxi 214122,China;State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学化学与材料工程学院,无锡214122 [2]江南大学食品科学与技术国家重点实验室,无锡214122

出　　处：《分析化学》2020年第6期750-756,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.51973083)资助。

摘　　要：建立了顶空-固相微萃取箭型-气相色谱-质谱(HS-SPME Arrow-GC-MS)联用技术,结合多变量统计分析铜绿微囊藻在模拟自然环境条件下,其挥发性代谢物组成及变化规律。通过选取合适萃取头,对不同时期铜绿微囊藻样品中挥发性代谢物进行定性分析,鉴定了210种挥发性代谢物。采用多变量统计分析方法,进一步研究铜绿微囊藻生长过程中挥发性代谢物的消长变化规律,筛选出10种统计学显著性差异代谢物:环己醇、二甲基三硫、苯甲醇、樟脑、2-甲氧基苯酚、3-己烯-1-醇、2, 4-癸二烯醛、吲哚、柠檬醛和正壬醇。优化了萃取温度、萃取时间、体系盐度等SPME Arrow萃取条件,筛选差异性代谢物的特征离子峰,并进行定量分析和方法学验证。结果表明,10种统计学显著性差异代谢物在0.050~1000 ng/L范围内具有良好的线性关系,相关系数>0.998,检出限范围为0.010~0.030 ng/L,回收率在76.3%~93.0%之间,RSD≤12.7%(n=6)。本方法操作简便、快速,灵敏度高,稳定性好,可用于蓝藻水华爆发初期天然水体中挥发性代谢标志物测定。A novel headspace solid phase microextraction arrow(HS-SPME Arrow) method combined with gas chromatography-mass spectrometry(GC-MS) was developed for extraction and analysis of volatile organic compounds of Microcystis aeruginosa under simulated natural conditions. By selecting suitable extraction fibers, 210 kinds of volatile metabolites were identified by full scanning qualitative analysis from the samples of Microcystis aeruginosa in producing period. The metabolic changes of volatile molecules during the growth of Microcystis aeruginosa were analyzed by multivariate statistical analysis and clustering analysis. Chemometric analysis based on principal component analysis, partial least squares-discriminant analysis and heatmap associated with hierarchical cluster analysis provided a suitable tool to differentiate volatile organic compounds in algal material. And 10 metabolites in statistical significance were identified, including cyclohexanol, dimethyl trisulfide, benzenemethanol, camphor, 2-methoxyphenol, 3-hexene-1-ol, 2,4-decadienal, indole, citral and 1-nonanol. Parameters affecting the extraction, such as extraction temperature, time and salt were carefully optimized. In addition, the analysis conditions, including desorption temperature and time as well as gas chromatographic parameters, were optimized. The calibration curves showed a good linearity(correlation coefficient > 0.998) in the concentration range of 0.050-1000 ng/L. The detection limits of this method were 0.010-0.030 ng/L, and the recoveries at the concentration level of 100 ng/L were 76.3%-3.0%. The method was satisfactorily precise, with the RSDs of less than 12.7%. The new method was simple, quick, stable, and applicable to complex matrices. It was suitable to the determination of volatile metabolites in natural water at the early stage of blue-green algae bloom.

关 键 词：顶空-固相微萃取箭型 气相色谱-质谱联用 多变量统计分析 铜绿微囊藻 挥发性代谢物 

分 类 号：X832[环境科学与工程—环境工程] O657.63[理学—分析化学]