Roth Simon R, Henkel Kerstin, Altenburger Markus J, Auwärter Volker, Neukamm Merja A
Institute of Forensic Medicine, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Albertstraße 9, 79104 Freiburg, Germany.
Hermann Staudinger Graduate School, University of Freiburg, Hebelstaße 27, 79104 Freiburg, Germany.
Bioanalysis. 2018 Aug 1;10(16):1319-1333. doi: 10.4155/bio-2018-0106. Epub 2018 Aug 24.
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% (w/v), 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.
磷脂脂肪酸甲酯(FAME)分析为表征微生物群落和监测变化提供了一种简单的方法,可作为16S rRNA测序的补充。建立了一种通过FAME谱表征牙菌斑的方法。
脂肪酸被测定为FAMEs(直接酸性酯交换法),并使用优化的温度梯度通过气相色谱-质谱联用仪(GC-MS)进行分析。使用分数因子中心复合面心设计优化酯交换反应。
甲醇/甲苯中酯交换的最佳条件:盐酸浓度2%(w/v),反应时间40分钟,温度110°C。方法验证显示出令人满意的准确度、精密度和线性。
该方法为通过FAME谱表征菌斑提供了一种有用的工具,并成功应用于来自十名受试者的样本,证明了其适用性。