Analysis of volatile metabolites from in vitro biofilms of Pseudomonas aeruginosa with thin-film microextraction by thermal desorption gas chromatography-mass spectrometry
Analysis of volatile metabolites from in vitro biofilms of Pseudomonas aeruginosa with thin-film microextraction by thermal desorption gas chromatography-mass spectrometry
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薄膜微萃取热解吸气相色谱-质谱法分析铜绿假单胞菌体外生物膜的挥发性代谢物
DOI：
10.1007/s00216-020-02529-4
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发表时间：
2020
影响因子：
4.3
通讯作者：
Schmitz O. J.
中科院分区：
文献类型：
--
作者：
Koehler T;Ackermann I;Brecht D;Uteschil F;Wingender J;Telgheder U;Schmitz O. J.
囊性纤维化是一种常染色体隐性遗传性疾病，导致呼吸道粘液增厚。这些条件有利于多种微生物的感染，如慢性肺部感染，其中铜绿假单胞菌是CF患者生命末期在肺内定植的主要病原菌。这项体外研究使用AP。部分囊性纤维化条件下的铜绿假单胞菌模型，细胞外挥发性代谢物取样。气体采样采用薄膜微萃取(TFME)和商用聚二甲基硅氧烷(PDMS)膜，负载膜的分析采用气相色谱-四极杆质谱联用热解吸法(TD-GC-QMS)。为此，对两种商用薄膜进行了热重分析，并与常压光电离质谱(TG-APPI-QMS)联用，研究了薄膜的均匀性和温度稳定性。使用本研究中开发的方法对选定的薄膜进行清洗。TD-GC-QMS方法被成功地用于BYP产生的挥发性代谢物的标准。铜绿假单胞菌。在低纳摩尔范围内(0.5nM和1.5nM)实现了对中低极性化合物的检测和定量。最后将所建立的方法应用于研究菌株P的生物膜产生的胞外挥发性代谢产物。好氧和厌氧条件下的铜绿假单胞菌DSM 50071。综上所述，在两种条件下都能检测到11种代谢物。此外，本研究还表明，不同的氧气条件(好氧和厌氧)会导致不同的胞外挥发性代谢产物的释放。可以确定特定的代谢物，如1-十一烯(需氧)和2-十一酮(需氧)。结果表明，生物被膜模型可能适用于OFP的鉴定。铜绿假单胞菌在临床条件下。此外，该模型可以作为研究不同细菌单独或联合培养的胞外挥发性代谢物的基础，以及像在CF肺中那样的肺部疾病的实施。这种可能性使非侵入性“床边”呼吸分析方法的开发成为可能，可用于各种细菌感染部位的CF患者。
Cystic fibrosis (CF) is an autosomal recessive inherited disease which leads to a production of thickened mucus in the airways. These conditions are conducive to poly-microbial infections, like chronic lung infection, in whichPseudomonas aeruginosa(P. aeruginosa) is the major pathogenic bacterium colonizing CF lungs at the end of the lifetime of CF patients. This in vitro study uses aP. aeruginosabiofilm model under partly cystic fibrosis conditions, with a sampling of volatile extracellular metabolites. The gas sampling was done with thin-film microextraction (TFME) and commercial polydimethylsiloxane (PDMS) films, whereas the analysis of loaded films was done by gas chromatography coupled to quadrupole mass spectrometry and thermodesorption (TD-GC-qMS). For this purpose, two commercially available films were characterized by means of thermogravimetry coupled to a qMS with atmospheric pressure photo ionization (TG-APPI-qMS), regarding homogeneity and temperature stability. The selected film was cleaned using a method developed in this study. The TD-GC-qMS method was successfully used for standards of volatile metabolites which were known to be produced byP. aeruginosa. Limits of detection and quantification of the method for middle and less polar compounds in low nanomolar range (0.5 nM and 1.5 nM) were achieved. The developed method was finally applied to investigate the extracellular volatile metabolites produced by biofilms of the strainP. aeruginosaDSM 50071 under aerobic and anaerobic conditions. In sum, eleven metabolites could be found under both conditions. Furthermore, it was shown in this study that different oxygen conditions (aerobic and anaerobic) resulted in emitting different extracellular volatile metabolites. Specific metabolites, like 1-undecene (aerobic) and 2-undecanone (anaerobic), could be identified. The results are promising, in that the biofilm model may be applicable for the identification ofP. aeruginosaunder clinical conditions. Furthermore, the model could be the basis for studying extracellular volatile metabolites from different mono- or co-cultures of various bacteria, as well as the implementation of pulmonary conditions, like these in CF lungs. This possibility allows the development of a non-invasive “at-bedside” breath analysis method for CF patients in focus of various bacterial infections.Graphical abstract
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工程化学：Von der Synthese bis zur Anwendung
影响因子：
6.1
作者：
W. Kaiser
通讯作者：
W. Kaiser
DOI：
10.1007/s11306-018-1357-5
发表时间：
2018
期刊：
Metabolomics : Official journal of the Metabolomic Society
影响因子：
--
作者：
Lawal O;Knobel H;Weda H;Nijsen TME;Goodacre R;Fowler SJ;BreathDx consortium
通讯作者：
BreathDx consortium
影响因子：
2.4
作者：
Savelev, S. U.;Perry, J. D.;De Soyza, A.
通讯作者：
De Soyza, A.
DOI：
10.1007/bf00454145
发表时间：
1956-01-01
期刊：
FRESENIUS ZEITSCHRIFT FUR ANALYTISCHE CHEMIE
影响因子：
--
作者：
KAISER, H;SPECKER, H
通讯作者：
SPECKER, H
影响因子：
8.8
作者：
Scholler, C;Molin, S;Wilkins, K
通讯作者：
Wilkins, K