Chemometric variance analysis of 1H NMR metabolomics data on the effects of oral rinse on saliva

Saliva is an easy to obtain bodily fluid that is specific to the oral environment. It can be used for metabolomic studies as it is representative of the overall wellbeing of an organism, as well as mouth health and bacterial flora. The metabolomic structure of saliva varies greatly depending on the bacteria present in the mouth as they produce a range of metabolites. In this study we have investigated the metabolomic profiles of human saliva that were obtained using ¹H NMR (nuclear magnetic resonance) analysis. 48 samples of saliva were collected from 16 healthy subjects over 3 days. Each sample was split in two and the first half treated with an oral rinse, while the second was left untreated as a control sample. The 96 ¹H NMR metabolomic profiles obtained in the dataset are affected by three factors, namely 16 subjects, 3 sampling days and 2 treatments. These three factors contribute to the total variation in the dataset. When analysing datasets from saliva using traditional methods such as PCA (principal component analysis), the overall variance is dominated by subjects’ contributions, and we cannot see trends that would highlight the effect of specific factors such as oral rinse. In order to identify these trends, we used methods such as MSCA (multilevel simultaneous component analysis) and ASCA (ANOVA simultaneous component analysis), that provide variance splits according to the experimental factors, so that we could look at the particular effect of treatment on saliva. The analysis of the treatment effect was enhanced, as it was isolated from the overall variance and assessed without confounding factors.

     

Chemometric variance analysis of <Superscript>1</Superscript>H NMR metabolomics data on the effects of oral rinse on saliva

In order to investigate exposure end effect markers of fruit and fruit fibre intake we investigated how fresh apple or apple-pectin affects the urinary metabolome of rats. Twenty-four Fisher 344 male rats were randomized into three groups and fed a standard diet with different supplementations added in two of the groups (7% apple-pectin or 10 g raw apple). After 24 days of feeding, 24 h urine was collected and analyzed by UPLC-QTOF-MS in positive and negative ionization mode. Metabolites that responded to the apple or pectin diets were selected and classified as either potential exposure or effect markers based on the magnitude and pattern of their response. An initial principal component analysis (PCA) of all detected metabolites showed a clear separation between the groups and during marker identification several new apple and/or pectin markers were found. Quinic acid, m-coumaric acid and (-)epicatechin were identified as exposure markers of apple intake whereas hippuric acid behaved as an effect marker. Pyrrole-2-carboxylic acid and 2-furoylglycine behaved as pectin exposure markers while 2-piperidinone was recognized as a potential pectin effect marker. None of them has earlier been related to intake of pectin or other fibre products. We discuss these new potential exposure and effect markers and their interpretation.