A compendium of all the volatile organic compounds (VOCs) emanating from the human body (the volatolome) is for the first time reported. 1840 VOCs have been assigned from breath (872), saliva (359), blood (154), milk (256), skin secretions (532) urine (279), and faeces (381) in apparently healthy individuals. Compounds were assigned CAS registry numbers and named according to a common convention where possible. The compounds have been grouped into tables according to their chemical class or functionality to permit easy comparison. Some clear differences are observed, for instance, a lack of esters in urine with a high number in faeces. Careful use of the database is needed. The numbers may not be a true reflection of the actual VOCs present from each bodily excretion. The lack of a compound could be due to the techniques used or reflect the intensity of effort e.g. there are few publications on VOCs from blood compared to a large number on VOCs in breath. The large number of volatiles reported from skin is partly due to the methodologies used, e.g. collecting excretions on glass beads and then heating to desorb VOCs. All compounds have been included as reported (unless there was a clear discrepancy between name and chemical structure), but there may be some mistaken assignations arising from the original publications, particularly for isomers. It is the authors' intention that this database will not only be a useful database of VOCs listed in the literature, but will stimulate further study of VOCs from healthy individuals. Establishing a list of volatiles emanating from healthy individuals and increased understanding of VOC metabolic pathways is an important step for differentiating between diseases using VOCs.

https://iopscience.iop.org/article/10.1088/1752-7155/8/1/014001/pdf

A review of the volatiles from the healthy human body

Recent advances in lactic acid production by microbial fermentation processes

Flavour development in dairy fermentations, most notably cheeses, results from a series of (bio)chemical processes in which the starter cultures provide the enzymes. Particularly the enzymatic degradation of proteins (caseins) leads to the formation of key-flavour components, which contribute to the sensory perception of dairy products. More specifically, caseins are degraded into peptides and amino acids and the latter are major precursors for volatile aroma compounds. In particular, the conversion of methionine, the aromatic and the branched-chain amino acids are crucial. A lot of research has focused on the degradation of caseins into peptides and free amino acids, and more recently, enzymes involved in the conversion of amino acids were identified. Most data are generated on Lactococcus lactis, which is the predominant organism in starter cultures used for cheese-making, but also Lactobacillus, Streptococcus, Propionibacterium and species used for surface ripening of cheeses are characterised in their flavour-forming capacity. In this paper, various enzymes and pathways involved in flavour formation will be highlighted and the impact of these findings for the development of industrial starter cultures will be discussed.

Flavour formation by lactic acid bacteria and biochemical flavour profiling of cheese products

Key odorants in various cheese types as determined by gas chromatography-olfactometry

Here, we review what is known about the microorganisms present in raw milk, including milk from cows, sheep, goats and humans. Milk, due to its high nutritional content, can support a rich microbiota. These microorganisms enter milk from a variety of sources and, once in milk, can play a number of roles, such as facilitating dairy fermentations (e.g. Lactococcus, Lactobacillus, Streptococcus, Propionibacterium and fungal populations), causing spoilage (e.g. Pseudomonas, Clostridium, Bacillus and other spore-forming or thermoduric microorganisms), promoting health (e.g. lactobacilli and bifidobacteria) or causing disease (e.g. Listeria, Salmonella, Escherichia coli, Campylobacter and mycotoxin-producing fungi). There is also concern that the presence of antibiotic residues in milk leads to the development of resistance, particularly among pathogenic bacteria. Here, we comprehensively review these topics, while comparing the approaches, both culture-dependent and culture-independent, which can be taken to investigate the microbial composition of milk.

The complex microbiota of raw milk

A new headspace-GC-sniffing method is proposed. Using a recently developed headspace cell, the vapor phase is collected under conditions that mimic well those of an aroma above a food. Data treatment is based on detection frequency, rather than on perceived intensity or successive dilutions as used in other approaches. Repeatability appears satisfactory, and independent panels are even able to generate similar aromagrams, without training prior to the analysis. Using a minimum of six assessors, this technique seems to be more reliable than classical ones. To compare detection frequencies between two aromagrams, an estimation of the least significant difference is given. A theoretical justification of this approach is suggested, on the basis of determination of detection thresholds. Keywords: Headspace-GC-sniffing; impact odorants; detection frequency

Hyphenated Headspace-Gas Chromatography-Sniffing Technique: Screening of Impact Odorants and Quantitative Aromagram Comparisons

Yogurt exhibits a delicate and low intense flavor which requires mild sample isolation techniques and sensitive identification means. This paper decribes the application of two new flavor separation techniques:  combined static and dynamic headspace, and preparative simultaneous distillation−extraction under vacuum. Of the 91 components identified by mass spectrometry, 21 were found to have a major impact on the aroma using a new method for the treatment of GC−sniffing data. One of these compounds, 1-nonen-3-one, was identified by spectral means for the first time in a flavor. Its odor detection threshold of 8 pg/kg appears to be one of the lowest among known flavor compounds. On the basis of the peak areas of yogurt and milk aromagrams, some compounds seem to be already present in milk, others appear to be generated by the strains. Keywords: Yogurt aroma; headspace; GC−olfactometry; impact-compound; composition

Determination and Origin of the Aroma Impact Compounds of Yogurt Flavor

The sensory properties of traditional acidic and mild, less acidic yogurts were characterized by a trained panel using a descriptive approach Many of the descriptive attributes varied almost linearly with pH, showing either a positive or negative correlation with increasing acidity The panel was very sensitive to acidity differences, as demonstrated by the linear relationship between acidity perception and pH Important flavor differences were found between the two classes of yogurt They were mainly due to differences in acidity and not to different concentrations of the three impact aroma compounds, acetaldehyde, 2,3-butanedione, and 2,3-pentanedione This emphasizes the importance of acidity in yogurt flavor Deodorization and impact aroma compound addition had much less influence on yogurt flavor than pH variations

Sensory investigation of yogurt flavor perception: mutual influence of volatiles and acidity.

A quick headspace GC method for quantification of volatiles was developed, involving only minor sample preparation. Yogurt flavor compounds could be quantified in the micrograms per kilogram to milligrams per kilogram range without any difficulty, despite the complex matrix. Volatiles of traditional acidic and mild, less acidic yogurts were compared, and important differences were found for acetaldehyde, 2,3-butanedione, and 2,3-pentanedione. Concentrations of 2,3-butanedione and 2,3-pentanedione increased 2-3-fold in mild, less acidic yogurts compared to traditional acidic ones. This is due to accumulation of the precursors of the diketones, 2-acetolactate and 2-acetohydroxybutyrate, during fermentation in mild, less acidic yogurt. These precursors are subsequently converted to the corresponding diketones during storage. On the contrary, acetaldehyde formation was reduced in the mild yogurt, due to growth differences between the lactic acid bacteria used for fermentation of the milk. The quantitative results presented in this study validate previous GC sniffing conclusions (Ott et al. J. Agric. Food Chem. 1997, 45, 850-858), showing that yogurt aroma is the superposition of impact flavor compounds generated by fermentation on milk compounds.

Aroma comparisons of traditional and mild yogurts: headspace gas chromatography quantification of volatiles and origin of alpha-diketones.

Lactobacillus johnsonii La1, a probiotic bacterium with demonstrated health effects, grows in milk, where it ferments lactose to D- and L-lactate in a 60:40% ratio. The D-lactate dehydrogenase (D-LDH) gene (ldhD) of this strain was isolated, and an in vitro-truncated copy of that gene was used to inactivate the genomic copy in two strains, La1 and N312, by gene replacement. For that, an 8-bp deletion was generated within the cloned ldhD gene to inactivate its function. The plasmid containing the altered ldhD was transferred to L. johnsonii via conjugative comobilization with Lactococcus lactis carrying pAMbeta1. Crossover integrations of the plasmid at the genomic ldhD site were selected, and appropriate resolution of the cointegrate structures resulted in mutants that had lost the plasmid and in which the original ldhD was replaced by the truncated copy. These mutants completely lacked D-LDH activity. Nevertheless, the lower remaining L-LDH activity of the cells was sufficient to reroute most of the accumulating pyruvate to L-lactate. Only a marginal increase in production of the secondary end products acetaldehyde, diacetyl, and acetoin was observed. It can be concluded that in L. johnsonii D- and L-LDH are present in substantial excess for their role to eliminate pyruvate and regenerate NAD(+) and that accumulated pyruvate is therefore not easily redirected in high amounts to secondary metabolic routes.

Andreas Ott

Papers

Hyphenated Headspace-Gas Chromatography-Sniffing Technique: Screening of Impact Odorants and Quantitative Aromagram Comparisons

Determination and Origin of the Aroma Impact Compounds of Yogurt Flavor

Sensory investigation of yogurt flavor perception: mutual influence of volatiles and acidity.

Aroma comparisons of traditional and mild yogurts: headspace gas chromatography quantification of volatiles and origin of alpha-diketones.

D-Lactate dehydrogenase gene (ldhD) inactivation and resulting metabolic effects in the Lactobacillus johnsonii strains La1 and N312.