RESUMO
The characterization of volatile compounds in biological fluids offers a distinct approach to study the metabolic imprint of foods on the human metabolome, particularly to identify novel biomarkers of food intake (BFIs) that are not captured by classic metabolomics. Using a combination of dynamic headspace vacuum transfer In Trap extraction and gas chromatography coupled with mass spectrometry, we measured volatile compounds (the "volatilome") in plasma and urine samples from a randomized controlled crossover intervention study in which 11 healthy subjects ingested milk, cheese, or a soy-based drink. More than 2000 volatile compounds were detected in plasma, while 1260 compounds were detected in urine samples. A postprandial response in plasma was confirmed for 697 features. Univariate and multivariate analyses identified four molecules in plasma and 31 molecules in urine samples differentiating the ingestion of the foods, of which three metabolites in plasma and nine in urine were specific to the dairy products. Among these molecules, heptan-2-one, 3,5-dimethyloctan-2-one, and undecan-2-one in plasma and 3-ethylphenol, heptan-2-one, 1-methoxy-2-propyl acetate, and 9-decenoic acid were highly discriminative for dairy or cheese intake. In urine, 22 volatile compounds were highly discriminative for soy-based drink intake. The majority of these molecules have not been reported in humans. Our findings highlight the potential of plasma and urinary volatilomics for detection of novel dietary biomarkers.
Assuntos
Queijo , Biomarcadores , Queijo/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Metaboloma , Metabolômica , LeiteRESUMO
The identification of molecular biomarkers that can be used to quantitatively link dietary intake to phenotypic traits in humans is a key theme in modern nutritional research. Although dairy products (with and without fermentation) represent a major food group, the identification of markers of their intake lags behind that of other food groups. Here, we report the results from an analysis of the metabolites in postprandial serum and urine samples from a randomized crossover study with 14 healthy men who ingested acidified milk, yogurt, and a non-dairy meal. Our study confirms the potential of lactose and its metabolites as markers of lactose-containing dairy foods and the dependence of their combined profiles on the fermentation status of the consumed products. Furthermore, indole-3-lactic acid and 3-phenyllactic acid are two products of fermentation whose postprandial behaviour strongly discriminates yogurt from milk intake. Our study also provides evidence of the ability of milk fermentation to increase the acute delivery of free amino acids to humans. Notably, 3,5-dimethyloctan-2-one also proves to be a specific marker for milk and yogurt consumption, as well as for cheese consumption (previously published data). These molecules deserve future characterisation in human interventional and observational studies.
Assuntos
Intolerância à Lactose , Leite , Masculino , Humanos , Animais , Leite/química , Iogurte , Lactose/análise , Estudos Cross-Over , Intolerância à Lactose/metabolismoRESUMO
This work aimed to determine the formation over time of 3-methylbutanal and 3-methylbutan-1-ol recognized as malty during the manufacture of Raclette-type cheese and the fermention of reconstituted skim milk, and filter-sterilized MRS broth. Using dynamic headspace-vacuum transfer in trap extraction followed by gas chromatography coupled with mass spectrometry-olfactometry (DHS-VTT-GC-MS-O) as a screening method for the malty compounds, five compounds (2-methylpropanal, 2- and 3-methylbutanal, and 2- and 3-methylbutan-1-ol) were identified as potential compounds causing the malty aroma in starter culture development and Raclette-type cheeses. Focus on compounds having a predominant sensorial effect (3-methylbutanal and 3-methylbutan-1-ol), spikings of leucine, 13C-labeled leucine, α-ketoisocaproic acid, and α-ketoglutaric acid provided a better understanding of their formation pathway. This study highlighted the discrepancies in the formation of 3-methylbutanal and 3-methylbutan-1-ol between the growth media; namely, despite the presence of free leucine available in MRS and the addition of an excess, no increase of the target compounds was observed. The concentration of these compounds in MRS increased only when α-ketoglutaric acid or α-ketoisocaproic acid was added, and a preference for the pathway to α-hydroxyisocaproic acid instead of 3-methylbutanal was shown. In addition, a formation of 3-methylbutanal when the bacteria were not yet active was observed when spiking α-ketoisocaproic acid, which potentially indicates that this part of the metabolism could take place extracellularly. These results could potentially unveil other, not-yet-identified reactants, directly influencing the production of compounds responsible for the malty aroma in Raclette cheese.
Assuntos
Aldeídos/metabolismo , Queijo/análise , Leite/química , Odorantes/análise , Animais , Bovinos , Queijo/microbiologia , Fermentação , Cromatografia Gasosa-Espectrometria de Massas , Lactobacillales/metabolismo , Leite/microbiologia , Olfatometria , SuíçaRESUMO
Headspace in-tube extraction (HS-ITEX) and solid phase microextraction (HS-SPME) sampling, followed by gas chromatography-mass spectrometry (GC-MS), are widely used to analyze volatile compounds in various food matrices. While the extraction efficiency of volatile compounds from foodstuffs is crucial for obtaining relevant results, these efficiency of these extraction methods limited by their long extraction times and requirements for large sample quantity. This study reports on the development and application of a new extraction technique based on HS-ITEX hardware, which improves the extraction rate and capacity by operating under reduced pressure, called Dynamic Headspace Vacuum Transfer In-Trap Extraction (DHS-VTT). The results of the study indicate that DHS-VTT improves the extraction of the target compounds. The area of the mass spectrometer signal for each compound can be up to 450 times more intense than the HS-SPME and HS-ITEX techniques performed in the same experimental conditions of extraction temperature and time. DHS-VTT runs in automated mode, making it possible to work with smaller sample quantity and also favors the HS extraction of all volatile compounds. In addition, the necessary modifications to the installation were cheap and the life of an ITEX trap is up to 10 times longer than an SPME fibre.