HILIC-IM-MS for Simultaneous Lipid and Metabolite Profiling of Bacteria

IF 4.6 Q1 CHEMISTRY, ANALYTICAL
Jana M. Carpenter, Hannah M. Hynds, Kingsley Bimpeh and Kelly M. Hines*, 
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引用次数: 0

Abstract

Although MALDI-ToF platforms for microbial identifications have found great success in clinical microbiology, the sole use of protein fingerprints for the discrimination of closely related species, strain-level identifications, and detection of antimicrobial resistance remains a challenge for the technology. Several alternative mass spectrometry-based methods have been proposed to address the shortcomings of the protein-centric approach, including MALDI-ToF methods for fatty acid/lipid profiling and LC-MS profiling of metabolites. However, the molecular diversity of microbial pathogens suggests that no single “ome” will be sufficient for the accurate and sensitive identification of strain- and susceptibility-level profiling of bacteria. Here, we describe the development of an alternative approach to microorganism profiling that relies upon both metabolites and lipids rather than a single class of biomolecule. Single-phase extractions based on butanol, acetonitrile, and water (the BAW method) were evaluated for the recovery of lipids and metabolites from Gram-positive and -negative microorganisms. We found that BAW extraction solutions containing 45% butanol provided optimal recovery of both molecular classes in a single extraction. The single-phase extraction method was coupled to hydrophilic interaction liquid chromatography (HILIC) and ion mobility-mass spectrometry (IM-MS) to resolve similar-mass metabolites and lipids in three dimensions and provide multiple points of evidence for feature annotation in the absence of tandem mass spectrometry. We demonstrate that the combined use of metabolites and lipids can be used to differentiate microorganisms to the species- and strain-level for four of the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa) using data from a single ionization mode. These results present promising, early stage evidence for the use of multiomic signatures for the identification of microorganisms by liquid chromatography, ion mobility, and mass spectrometry that, upon further development, may improve upon the level of identification provided by current methods.

Abstract Image

Abstract Image

HILIC-IM-MS同时分析细菌的脂质和代谢物
尽管用于微生物鉴定的MALDI-ToF平台在临床微生物学领域取得了巨大成功,但仅使用蛋白质指纹图谱来区分近缘物种、菌株水平鉴定和抗微生物药物耐药性检测仍然是该技术的挑战。已经提出了几种基于质谱的替代方法来解决以蛋白质为中心的方法的缺点,包括用于脂肪酸/脂质分析的MALDI-ToF方法和代谢物的LC-MS分析。然而,微生物病原体的分子多样性表明,没有一个单一的“基因组”足以准确和敏感地识别细菌的菌株和敏感水平谱。在这里,我们描述了一种微生物分析的替代方法的发展,这种方法依赖于代谢物和脂质,而不是单一种类的生物分子。以丁醇、乙腈和水为基础的单相萃取法(BAW法)对革兰氏阳性和阴性微生物的脂质和代谢物的回收率进行了评估。我们发现含有45%丁醇的BAW提取液在一次提取中对这两类分子的回收率最佳。将单相萃取法与亲水性相互作用液相色谱法(HILIC)和离子迁移质谱法(IM-MS)相结合,对相似质量的代谢物和脂质进行三维解析,并在没有串联质谱法的情况下为特征注释提供多点证据。我们证明,使用单一电离模式的数据,代谢物和脂质可以用于区分四种ESKAPE病原体(屎肠球菌、金黄色葡萄球菌、鲍曼不动杆菌和铜绿假单胞菌)的种和菌株水平的微生物。这些结果为利用多组特征通过液相色谱、离子迁移率和质谱鉴定微生物提供了有希望的早期证据,在进一步发展的基础上,可能会提高当前方法提供的鉴定水平。
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来源期刊
ACS Measurement Science Au
ACS Measurement Science Au 化学计量学-
CiteScore
5.20
自引率
0.00%
发文量
0
期刊介绍: ACS Measurement Science Au is an open access journal that publishes experimental computational or theoretical research in all areas of chemical measurement science. Short letters comprehensive articles reviews and perspectives are welcome on topics that report on any phase of analytical operations including sampling measurement and data analysis. This includes:Chemical Reactions and SelectivityChemometrics and Data ProcessingElectrochemistryElemental and Molecular CharacterizationImagingInstrumentationMass SpectrometryMicroscale and Nanoscale systemsOmics (Genomics Proteomics Metabonomics Metabolomics and Bioinformatics)Sensors and Sensing (Biosensors Chemical Sensors Gas Sensors Intracellular Sensors Single-Molecule Sensors Cell Chips Arrays Microfluidic Devices)SeparationsSpectroscopySurface analysisPapers dealing with established methods need to offer a significantly improved original application of the method.
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