Determination of porphyrins in oral bacteria by liquidchromatography electrospray ionization tandemmass spectrometry
2015 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650Article in journal (Refereed) Published
Biofilms in the oral cavity can be visualized byfluorescence and a common assumption is that the endogenouslyproduced porphyrins in certain bacteria give rise tothis fluorescence. Porphyrin content in oral bacteria hasbeen sparingly investigated, and non-selective detectiontechniques such as utilizing the Soret fluorescence bandof porphyrins are often used. In the present study, a quantitativeand selective method for the determination of porphyrinsin oral bacteria has been developed and validatedusing high performance liquid chromatography-tandemmass spectrometry. Lysis of bacteria using Tris-EDTA buffertogether with ultrasonication showed high microbial killingefficiency ≥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 contentwas determined in the two oral pathogens Aggregatibacteractinomycetemcomitans and Porphyromonas gingivalis, aswell as for baker’s yeast, Saccharomyces cerevisiae.Uroporphyrin, 7-carboxylporphyrin, 6-carboxylporphyrin,coproporphyrin, and protoporphyrin IX were identified inthe investigated microorganisms, and it was shown that theporphyrin profile differs between the two bacteria, as wellas for S. cerevisiae. To our knowledge, this is the firsttime the porphyrin profile has been determined for thebacterium A. actinomycetemcomitans.
Place, publisher, year, edition, pages
HPLC/MS/MS, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Saccharomyces cerevisiae, Porphyrins, Oral bacteria
Research subject Analytical Chemistry
IdentifiersURN: urn:nbn:se:su:diva-135378DOI: 10.1007/s00216-015-8864-2OAI: oai:DiVA.org:su-135378DiVA: diva2:1045083
FunderSwedish Research Council, K2014-70X -22533-01-3