Determination of porphyrins in oral bacteria by liquid chromatography electrospray ionization tandem mass spectrometry.
Determination of porphyrins in oral bacteria by liquid chromatography electrospray ionization tandem mass spectrometry.
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DOI：
10.1007/s00216-015-8864-2
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发表时间：
2015-09
影响因子：
4.3
通讯作者：
Östman C
中科院分区：
文献类型：
--
作者：
Fyrestam J;Bjurshammar N;Paulsson E;Johannsen A;Östman C
口腔中的生物膜可以通过荧光可视化，并且常见的假设是某些细菌中内源性产生的卟啉引起这种荧光。口腔细菌中卟啉含量的研究很少，通常使用非选择性检测技术，如利用卟啉的Soret荧光带。在本研究中，已经开发和验证了使用高效液相色谱-串联质谱定量和选择性的方法测定口腔细菌中的卟啉。使用Tris-EDTA缓冲液裂解细菌结合超声处理显示出较高的微生物杀灭效率≥ 99.98%，并且使用C18-固相萃取净化样品导致所有分析物的低基质效应≤ 14%。使用这种方法，卟啉含量测定在两个口腔病原体Aggregatibacteractinomycetemcomitans和牙龈卟啉单胞菌，以及面包酵母，酿酒酵母。在所研究的微生物中鉴定了尿卟啉、7-羧基卟啉、6-羧基卟啉、粪卟啉和原卟啉IX，并且表明两种细菌之间以及S.啤酒。据我们所知，这是首次确定细菌A的卟啉谱。伴放线菌口腔细菌中卟啉的测定。本文的在线版本（doi：10.1007/s 00216 -015-8864-2）包含补充材料，可供授权用户使用。
Biofilms in the oral cavity can be visualized by fluorescence and a common assumption is that the endogenously produced porphyrins in certain bacteria give rise to this fluorescence. Porphyrin content in oral bacteria has been sparingly investigated, and non-selective detection techniques such as utilizing the Soret fluorescence band of porphyrins are often used. In the present study, a quantitative and selective method for the determination of porphyrins in oral bacteria has been developed and validated using high performance liquid chromatography-tandem mass spectrometry. Lysis of bacteria using Tris-EDTA buffer together with ultrasonication showed high microbial killing efficiency ≥99.98 %, and sample clean-up using C18-solid phase extraction resulted in low matrix effects ≤14 % for all analytes. Using this method, the porphyrin content was determined in the two oral pathogens Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, as well as for baker’s yeast, Saccharomyces cerevisiae. Uroporphyrin, 7-carboxylporphyrin, 6-carboxylporphyrin, coproporphyrin, and protoporphyrin IX were identified in the investigated microorganisms, and it was shown that the porphyrin profile differs between the two bacteria, as well as for S. cerevisiae. To our knowledge, this is the first time the porphyrin profile has been determined for the bacterium A. actinomycetemcomitans. Determination of porphyrins in oral bacteria. The online version of this article (doi:10.1007/s00216-015-8864-2) contains supplementary material, which is available to authorized users.