Volatile organic compound markers of psychological stress in skin: a pilot study.

The forehead was studied as a possible sampling site for capturing changes in volatile organic compound (VOC) profiles associated with psychological-stress. Skin-VOCs were sampled with a polydimethylsilicone (PDMS)-coupon and the resulting VOCs were recovered and analysed with two-stage thermal desorption gas chromatography-mass spectrometry. Fifteen young adult volunteers (19 years-26 years) participated in two interventions run in a randomised crossover design. One intervention, termed 'Neutral', required the participants to listen to peaceful music, the other, termed a 'paced audio serial addition task', required the participants to undertake a series of rapid mental arithmetic calculations in a challenging environment that induced a stress response. Skin-VOC samples were taken during each intervention. The resultant data were processed with dynamic background compensation, deconvolved, and registered to a common retention index scale. The importance of freezing skin patch samplers to -80 °C was determined during the method development phase of this study. The cumulative distribution function of the GC-MS data indicates the possibility that PDMS-coupons are selective towards the lower volatility VOC components in skin. The frequency distribution of the GC-MS data was observed to be approximately log-normal, and on the basis of this study, a further two-orders of magnitude reduction in sensitivity may be required before the complete skin-VOC profile may be characterised. Multi-variate analysis involving Pareto-scaling prior to partial least squares discriminant analysis identified four VOCs with the highest probability of contributing to the variance between the two states, and the responses to these VOCs were modelled with principle components analysis (PCA). Two VOCs, benzoic acid and n-decanoic acid were upregulated (14 and 8 fold respectively) and appear to be PASAT sensitive, with areas under (AUC) their receiver operator characteristic (ROC) curves of 0.813 and 0.852 respectively. A xylene isomer and 3-carene were down regulated 75% and 97% respectively, and found to be predictive of the neutral intervention (ROC AUC values of 0.898 and 0.929 respectively). VOC profiles in skin appear to change with stress either due to increased elimination, elevated bacterial activity, or perhaps increased oxidative pathways.

Spatial variations in the microbial community structure and diversity of the human foot is associated with the production of odorous volatiles.

The human foot provides an ideal environment for the colonization and growth of bacteria and subsequently is a body site associated with the liberation of odour. This study aimed to enumerate and spatially map bacterial populations' resident across the foot to understand any association with odour production. Culture-based analysis confirmed that Staphylococci were present in higher numbers than aerobic corynebacteria and Gram-positive aerobic cocci, with all species being present at much higher levels on the plantar sites compared to dorsal sites. Microbiomic analysis supported these findings demonstrating that Staphylococcus spp. were dominant across different foot sites and comprised almost the entire bacterial population on the plantar surface. The levels of volatile fatty acids, including the key foot odour compound isovaleric acid, that contribute to foot odour were significantly increased at the plantar skin site compared to the dorsal surface. The fact that isovaleric acid was not detected on the dorsal surface but was present on the plantar surface is probably attributable to the high numbers of Staphylococcus spp. residing at this site. Variations in the spatial distribution of these microbes appear to be responsible for the localized production of odour across the foot.

High throughput volatile fatty acid skin metabolite profiling by thermal desorption secondary electrospray ionisation mass spectrometry.

The non-invasive nature of volatile organic compound (VOC) sampling from skin makes this a priority in the development of new screening and diagnostic assays. Evaluation of recent literature highlights the tension between the analytical utility of ambient ionisation approaches for skin profiling and the practicality of undertaking larger campaigns (higher statistical power), or undertaking research in remote locations. This study describes how VOC may be sampled from skin and recovered from a polydimethylsilicone sampling coupon and analysed by thermal desorption (TD) interfaced to secondary electrospray ionisation (SESI) time-of-flight mass spectrometry (MS) for the high throughput screening of volatile fatty acids (VFAs) from human skin. Analysis times were reduced by 79% compared to gas chromatography-mass spectrometry methods (GC-MS) and limits of detection in the range 300 to 900 pg cm(-2) for VFA skin concentrations were obtained. Using body odour as a surrogate model for clinical testing 10 Filipino participants, 5 high and 5 low odour, were sampled in Manilla and the samples returned to the UK and screened by TD-SESI-MS and TD-GC-MS for malodour precursors with greater than >95% agreement between the two analytical techniques. Eight additional VFAs were also identified by both techniques with chains 4 to 15 carbons long being observed. TD-SESI-MS appears to have significant potential for the high throughput targeted screening of volatile biomarkers in human skin.

Functional characterisation of a SNP in the ABCC11 allele - effects on axillary skin metabolism, odour generation and associated behaviours.

BACKGROUND: A single nucleotide polymorphism (SNP), 538G→A, leading to a G180R substitution in the ABCC11 gene results in reduced concentrations of apocrine derived axillary odour precursors. OBJECTIVE: Determine the axillary odour levels in the SNP ABCC11 genotype variants and to investigate if other parameters associated with odour production are affected. METHODS: Axillary odour was assessed by subjective quantification and gas chromatography headspace analysis. Metabolite profiles, microbiome diversity and personal hygiene habits were also assessed. RESULTS: Axillary odour in the A/A homozygotes was significantly lower compared to the G/A and G/G genotypes. However, the perception-based measures still detected appreciable levels of axillary odour in the A/A subjects. Metabolomic analysis highlighted significant differences in axillary skin metabolites between A/A subjects compared to those carrying the G allele. These differences resulted in A/A subjects lacking specific volatile odourants in the axillary headspace, but all genotypes produced odoriferous short chain fatty acids. Microbiomic analysis revealed differences in the relative abundance of key bacterial genera associated with odour generation between the different genotypes. Deodorant usage indicated a high level of self awareness of axillary odour levels with A/A individuals less likely to adopt personal hygiene habits designed to eradicate/mask its presence. CONCLUSIONS: The SNP in the ABCC11 gene results in lower levels of axillary odour in the A/A homozygotes compared to those carrying the G allele, but A/A subjects still produce noticeable amounts of axillary odour. Differences in axillary skin metabolites, bacterial genera and personal hygiene behaviours also appear to be influenced by this SNP.

Sampling and characterisation of volatile organic compound profiles in human saliva using a polydimethylsiloxane coupon placed within the oral cavity.

Evaluation of published methods reveals that existing methods for saliva sampling do not address the physical-chemical attributes of volatile organic compounds (VOCs). This study describes and presents evidence for adopting in situ sampling of salivary VOCs directly from the oral cavity using a polydimethylsiloxane (PDMS) based sampler. In vitro studies indicated that the vapour pressure of analytes was a factor in both the recovery of analytes and the precision of the recovery. The highest recoveries were observed for VOCs with the lowest vapour pressures, for example 5-nonanol (vapour pressure (P(v)) = 14 Pa) recoveries were approximately 20 times greater than those observed for octane (P(v) = 1726 Pa). Similarly, relative standard deviations reduced with vapour pressure, with the RSD for 5-nonanol responses observed to be 2.7% when compared to RSD = 26% for octane. Evaluation of VOCs recovered from 6 in vivo samples indicated that VOC concentrations in saliva may follow log-normal distributions; log-normal RSDs falling between 4.4% and 18.2% across the range of volatilities encountered. Increasing sampling time from 1 to 30 minutes indicated that the recovery of VOC into the sampler was affected by interaction between different physical-chemical properties and biogenic flux. A sampling time of 10 min was found to offer an acceptable compromise that enabled a representative sample to be acquired for the widest range of observed VOC behaviours with the sampler. The potential to 'tune' the sampling protocol for targeted analysis based on these factors was also noted. Comparison with passive drool saliva collection revealed up to 10(5) enhancement with reduced variability compared to drooled samples. This approach to in situ saliva sampling appears to have significant analytical utility for studying volatile signatures in humans.

Novel noninvasive identification of biomarkers by analytical profiling of chronic wounds using volatile organic compounds.

A complex profile of volatile organic compounds ("VOC"s) emanates from human skin, which is altered by changes in the body's metabolic or hormonal state, the external environment, and the bacterial species colonizing the skin surface. The aim of this study was to compare VOC profiles sampled from chronic leg wounds with those from asymptomatic skin. Five participants with chronic arterial leg ulcers were selected. VOC samples were obtained using polydimethylsilicone membranes ("skin-patch method") and analyzed by gas chromatography-ion trap mass spectrometry. Resultant data were analyzed using multivariate analysis and mass spectral matches were compared against the National Institute of Standards and Technology database. Principal component analysis showed differences in profiles obtained from healthy skin and boundary areas and between profiles from healthy skin and lesion samples (p<0.05). Partial least squares for discriminant analysis gave an average prediction accuracy of 73.3% (p<0.05). Mass spectral matching (verified against microbial swab results) identified unique VOCs associated with each sample area, wound bacterial colonization, and ingested medications. This study showcases a reproducible, robust, noninvasive methodology that is applicable in a clinical setting and may offer a new, hitherto unexplored, class of biochemical markers underpinning the metabolism of chronic wounds.