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基于硅胶管吸附萃取的热脱附-气相色谱-质谱联用法测定真菌的挥发性有机化合物
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科技基础性工作专项(2014FY120900);浙江大学实验技术研究项目(SJS201915)


Determination of fungal volatile organic compounds by thermal desorption-gas chromatography-mass spectrometry analysis based on silicone tube sorptive extraction
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    摘要:

    【背景】真菌代谢产生的挥发性有机化合物(volatile organic compounds,VOCs)具有重要的生物学功能。高通量的VOCs测定方法对于VOCs产生菌的筛选非常重要。【目的】建立一种简单可靠、基于硅胶管吸附萃取(silicone tube sorptive extraction,STSE)的热脱附-气相色谱-质谱(Thermal desorption-gas chromatography-mass spectrometry,TD-GC-MS)联用法测定真菌培养物VOCs的高通量分析技术。【方法】选择代表性的包括酯、酮、醇、醛、烯等21种VOCs作为对照品建立标准曲线;将对照品滴加于PDA培养基,基于STSE进行TD-GC-MS分析,测定其专属性;将木霉菌T552接种于PDA培养基,基于STSE进行TD-GC-MS分析,测定方法的重复性;在以上基础上,用上述试验条件对6株木霉菌产生的VOCs进行筛选。【结果】在全扫描和提取离子条件下,21种对照品分别在1.0?10.0 μg/mL、0.1?10.0 μg/mL范围内线性关系良好。基于STSE的TD-GC-MS分析发现,滴加在PDA培养基上的对照品分离良好,且PDA培养基平板中的其它物质不干扰VOCs的测定,表明该方法专属性良好;对PDA培养的木霉菌T552进行的VOCs重复测定鉴定的5种VOCs相对标准偏差均小于10%,表明该方法重复性良好。用建立的基于STSE的TD-GC-MS方法从6株供试木霉菌中鉴定出包括酸、醇、酯、烯等共37种VOCs。【结论】建立了一种简单可靠、基于STSE的TD-GC-MS分析测定真菌培养物VOCs的高通量分析技术,可用于真菌VOCs的分析测定。

    Abstract:

    [Background] Volatile organic compounds (VOCs) produced by fungi have important biological functions. The high-throughput VOCs analysis method is required for the screening of VOCs producing fungi. [Objective] To establish a robust, simple, high-throughput analysis technique for VOCs produced in petri dishes by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) analysis based on silicone tube sorptive extraction (STSE). [Methods] twenty-one representative VOCs including esters, ketones, alcohols, aldehydes and alkenes were selected as reference substances to establish the standard curve. TD-GC-MS analysis based on STSE was performed by dropping the reference substances into the PDA medium to determine the specificity and by inoculating Trichoderma strain T552 into the PDA medium to determine the repeatability. On this basis, the VOCs produced by 6 strains of Trichoderma were screened with the above method. [Results] The linear relationships of 21 reference substances were good in the range of 1.0?10.0 μg/mL and 0.1?10.0 μg/mL under the conditions of full scanning and extraction ion. TD-GC-MS analysis based on STSE showed that the reference substances dropped into the PDA separated well, and other substances in the PDA petri dish did not interfere with the determination of VOCs, indicating that the method had good specificity. Repeated determination of VOCs produced by Trichoderma strain T552 cultivated in the PDA medium was carried out, the relative standard deviation of the five VOCs identified were less than 10%, indicating that the method had good repeatability. A total of 37 VOCs including acid, alcohol, ester and alkene were identified from 6 Trichoderma strains were screened out by the established TD-GC-MS analysis based on STSE. [Conclusion] A robust, simple, high-throughput analysis technique for VOCs produced in petri dishes by TD-GC-MS analysis based on STSE was established, which can be used for the determination of fungal VOCs.

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毛黎娟,葛洋,周文武,祝增荣,章初龙. 基于硅胶管吸附萃取的热脱附-气相色谱-质谱联用法测定真菌的挥发性有机化合物[J]. 微生物学通报, 2019, 46(7): 1748-1758

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  • 在线发布日期: 2019-07-02
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