建立了一种测定热纤梭杆菌发酵液中的低相对分子质量有机酸的毛细管离子色谱-串联质谱方法.探索优化了色谱和质谱的检测条件,在最佳分析条件下同时检测8种有机酸.离子色谱以KOH水溶液作为流动相进行梯度洗脱;用IonSwift MAX-100毛细管柱进行分离;在喷雾电压为3.0 kV、喷雾气压强为 2 000 kPa、成源温度为275 ℃的条件下,选择离子监测(SIM)模式下运行质谱.结果表明,柠檬酸和异柠檬酸两种同分异构体能够得到很好的分离,8种有机酸在一定浓度范围内具有良好的线性关系,相对标准偏差为1.45%~5.99%,相关系数为0.9696~0.9986,平均加标回收率为89.0%~110.0%, 8种有机酸的检出限为0.01~0.50mg/L.该方法进样量少,灵敏度高,重现性好,能够满足实际样品的检测要求,可用于嗜热厌氧菌发酵液中低分子量有机酸的测定.
A capillary ion chromatography-mass spectrometry method was proposed to determine low relative molecular mass organic acids in culture media of Clostridium thermocellum.The conditions of analysis were optimized and eight organic acids were detected simultaneously.The target compounds were separated on an IonSwift MAX-100 capillary column with potassium hydroxide solution as mobile phase by gradient elution, and detected under selected ion monitoring (SIM) mode.The mass spectrometry conditions were: spray voltage 3.0 kV, sheath gas 2 000 kPa, capillary temperature 275 ℃.The relative standard deviations ranged from 1.45% to 5.99%.The coefficients of determination ranged from 0.9696 to 0.9986 and the mean recoveries were 89.0%-110.0%.The limits of detection of the eight organic acids were 0.01-0.50mg/L.This method was used to detect low relative molecular mass organic acids in culture media of Clostridium thermocellum with high sensitivity and good reproducibility.
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