Metabolites · 2025
Comprehensive GC/MS Profiling of Volatile Organic Compounds in Whole and Glandular Saliva Using MonoTrap Micro-Extraction.
СтатьяНазвание и аннотация: Перевод в очереди
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Mori Asuka, Kuboniwa Masae, Fukusaki Eiichiro
Аннотация
BACKGROUND/OBJECTIVES: Salivary volatile organic compounds (VOCs) are promising noninvasive biomarkers for a wide range of diseases. While glandular saliva, secreted by salivary glands, is a relatively pure biofluid, whole saliva is a complex mixture containing oral microbiota, food debris, and desquamated epithelial cells. Therefore, a comprehensive comparison of the VOC profiles of these two types of saliva is essential to identify biologically relevant compounds. In this study, we aimed to establish a reliable method for VOC profiling from small saliva volumes and identify VOCs that reflect the biological differences between glandular and whole saliva.
METHODS: We developed a protocol combining MonoTrap extraction with dichloromethane, allowing the analysis of VOCs from just 100 µL of saliva. To address the issue of sampling-derived artifacts, we implemented a two-step blank analysis to systematically exclude compounds originating from the collection device.
RESULTS: Our analysis successfully identified a total of 72 VOCs. Following blank analysis, we systematically excluded 15 artifacts originating from the sampling device. Subsequent orthogonal partial least squares discriminant analysis (OPLS-DA) and Wilcoxon signed-rank test (using variable importance for prediction (VIP) > 1.0 and q < 0.05) identified 10 key VOCs that were significantly higher in whole saliva than in glandular saliva. These compounds included isobutyric acid, isovaleric acid, 4-methylvaleric acid, 3-phenylpropionic acid, indole, skatole, methyl mercaptan, 1-propanol, δ-valerolactam, and acetaldehyde. Most of these compounds originate from the metabolic activities of the oral microbiome, suggesting that the distinct VOC profile of whole saliva is predominantly influenced by microbial activity.
CONCLUSIONS: Our findings demonstrated the effectiveness of this method for identifying biologically relevant VOCs from relatively small sample volumes. The identified VOC profiles highlight the contribution to the discovery of non-invasive biomarkers for oral health and serve as a solid foundation for future research into clinical applications.
METHODS: We developed a protocol combining MonoTrap extraction with dichloromethane, allowing the analysis of VOCs from just 100 µL of saliva. To address the issue of sampling-derived artifacts, we implemented a two-step blank analysis to systematically exclude compounds originating from the collection device.
RESULTS: Our analysis successfully identified a total of 72 VOCs. Following blank analysis, we systematically excluded 15 artifacts originating from the sampling device. Subsequent orthogonal partial least squares discriminant analysis (OPLS-DA) and Wilcoxon signed-rank test (using variable importance for prediction (VIP) > 1.0 and q < 0.05) identified 10 key VOCs that were significantly higher in whole saliva than in glandular saliva. These compounds included isobutyric acid, isovaleric acid, 4-methylvaleric acid, 3-phenylpropionic acid, indole, skatole, methyl mercaptan, 1-propanol, δ-valerolactam, and acetaldehyde. Most of these compounds originate from the metabolic activities of the oral microbiome, suggesting that the distinct VOC profile of whole saliva is predominantly influenced by microbial activity.
CONCLUSIONS: Our findings demonstrated the effectiveness of this method for identifying biologically relevant VOCs from relatively small sample volumes. The identified VOC profiles highlight the contribution to the discovery of non-invasive biomarkers for oral health and serve as a solid foundation for future research into clinical applications.