Multivariate optimization of a method for the determination of fatty acids in dental biofilm by GC-MS

• Published in: Bioanalysis Vol. 10; no. 16; pp. 1319 - 1333
• Main Authors: Roth, Simon R, Henkel, Kerstin, Altenburger, Markus J, Auwärter, Volker, Neukamm, Merja A
• Format: Journal Article
• Language: English
• Published: England Future Science Ltd 01.08.2018
• Subjects: Biofilms; dental plaque; Dental Plaque - microbiology; Esterification; fatty acids; Fatty Acids - analysis; Fatty Acids - chemistry; Gas Chromatography-Mass Spectrometry; lipids; method development; Multivariate Analysis; Reproducibility of Results; Temperature; toxicology; transesterification; validation; toxicology; lipids; fatty acids; transesterification; method development; dental plaque; validation
• ISSN: 1757-6180; 1757-6199
• DOI: 10.4155/bio-2018-0106

Phospholipid fatty acid methyl ester (FAME) analysis offers a simple option additionally to 16S rRNA sequencing to characterize microbial communities and to monitor changes. A method was established for the characterization of dental plaque via FAME profiles. Fatty acids were determined as FAMEs (direct, acidic transesterification) and analyzed by GC-MS using an optimized temperature gradient. The transesterification reaction was optimized using a fractional factorial central composite face-centered design. Optimal conditions for the transesterification in methanol/toluene: hydrochloric acid concentration 2% ( ), reaction time 40 min, temperature 110 °C. Method validation showed satisfactory accuracy, precision and linearity. The method provides a useful tool to characterize plaque via FAME profiles and was successfully applied to samples from ten subjects demonstrating its applicability.