Revisiting gas-chromatography/mass-spectrometry molar response factors for quantitative analysis (FID or TIC) of glycosidic linkages in polysaccharides produced by oral bacterial biofilms

IF 1.7 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2024-05-03 DOI:10.1016/j.mimet.2024.106942

Alexander J. Kendall, Polliana M.C. Scaffa, Matthew G. Logan, Carmem S. Pfeifer

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引用次数: 0

Abstract

Methylation analysis was performed on methylated alditol acetate standards and Streptococcus mutans extracellular polymeric substances (EPS) produced from wild-type and Gtf knockout strains (∆GtfB, ∆GtfB, and ∆GtfD). The methylated alditol acetate standards were representative of glycosidic linkages found in S. mutans EPS and were used to calibrate the GC–MS system for an FID detector and MS (TIC) and produce molar response factor, a necessary step in quantitative analysis. FID response factors were consistent with literature values (Sweet et al., 1975) and found to be the superior option for quantitative results, although the TIC response factors now give researchers without access to an FID detector a needed option for molar response factor correction. The GC–MS analysis is then used to deliver the ratio of the linkage types within a biofilm.

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重新审视用于定量分析（FID 或 TIC）口腔细菌生物膜产生的多糖中糖苷键的气相色谱/质谱摩尔响应因子。

对甲基化醛糖醇醋酸盐标准品以及野生型和 Gtf 基因敲除菌株（∆GtfB、∆GtfB 和 ∆GtfD）产生的变异链球菌胞外聚合物质（EPS）进行了甲基化分析。甲基化醛糖醇醋酸酯标准品是变异单胞菌 EPS 中发现的糖苷键的代表，用于校准 FID 检测器和 MS（TIC）的气相色谱-质谱系统，并产生摩尔响应因子，这是定量分析的必要步骤。FID 响应因子与文献值（Sweet 等人，1975 年）一致，被认为是定量结果的最佳选择，尽管 TIC 响应因子现在为无法使用 FID 检测器的研究人员提供了校正摩尔响应因子的必要选择。气相色谱-质谱（GC-MS）分析随后可用于提供生物膜内连接类型的比率。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.