Identification of aroma compounds in a commonly prescribed oral nutritional supplement and associated changes in olfactory abilities with human ageing.

Undernutrition is prevalent in the older adult population. Oral nutritional supplements (ONS) are a clinically effective nutritional intervention, however, patient acceptance of ONS can be limited by their palatability. While sensory attributes such as sweetness and mouthfeel have been investigated, the contribution made by aroma to the perceived flavour of ONS has not been studied. Firstly, this research aimed to identify the aroma active compounds within a commonly prescribed ONS using estimated odour activity values (OAV) and gas chromatography olfactometry mass spectrometry (GC-O-MS). Secondly, age related differences in olfactory detection were explored. Eight aroma active compounds were identified within the ONS, including diacetyl (sweet), isoamyl acetate (banana), dimethyl trisulfide (sulfur) and methanethiol (sulfur). When compared with younger adults (n = 24, 18-44 years), older adults (n = 24, 62-80 years) had higher detection thresholds for all aroma compounds and this was significant for isoamyl acetate (sweet, fruity) and methanethiol (sulfur) (p = 0.01 and p = 0.03, respectively). Thus, a decline in olfactory sensitivity was present in the older subjects included in the study, and this reduced detection sensitivity was aroma specific. Thus, older adults' flavour perception of ONS likely depends on the combined effect of product factors (the aroma profile) along with age related consumer factors (the degree of impairment in perception). This is a fundamental study which will aid future research into how the aroma profile, and associated age related impairments in perception, shape the global perception of ONS for nutritionally at risk older individuals.

Understanding the lost functionality of ethanol in non-alcoholic beer using sensory evaluation, aroma release and molecular hydrodynamics.

Consumer sensory evaluation, aroma release analysis and biophysical protein analysis were used to investigate the effect of ethanol on the release and perception of flavour in beer (lager and stout) at different ethanol levels (0 and 5% ABV). Consumer study results showed no significant differences in orthonasal perception, yet retronasal results showed that 0% lager was perceived as maltier with reduced fruitiness, sweetness, fullness/body and alcohol warming sensation (p < 0.05). Whilst ethanol alone decreases the aroma release regardless of LogP, the presence of α-amylase selectively reduces the headspace concentration of hydrophobic compounds. It was found that ethanol has a subtle inhibitory effect on the binding of hydrophobic compounds to α-amylase, thereby increasing their headspace concentration in the 5% ABV as compared to the 0% beers. This synergistic ethanol * saliva effect is attributed to the changes in the conformation of α-amylase due to ethanol-induced denaturation. It is hypothesised that the partially unfolded protein structures have a lower number of hydrophobic pockets, leading to a lower capacity to entrap hydrophobic aroma compounds. This supports the hypothesis that ethanol * saliva interactions directly impact the sensory and flavour properties of beer, which would provide a basis for further investigations in reformulation of 0% ABV drinks.

Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting.

Bone regeneration is a complex biological process where major cellular changes take place to support the osteogenic differentiation of mesenchymal bone progenitors. To characterise these biological changes and better understand the pathways regulating the formation of mature bone cells, the metabolic profile of mesenchymal stem cell (MSC) differentiation in vitro has been assessed non-invasively during osteogenic (OS) treatment using a footprinting technique. Liquid chromatography (LC)-mass spectrometry (MS)-based metabolite profiling of the culture medium was carried out in parallel to mineral deposition and alkaline phosphatase activity which are two hallmarks of osteogenesis in vitro. Metabolic profiles of spent culture media with a combination of univariate and multivariate analyses investigated concentration changes of extracellular metabolites and nutrients linked to the presence of MSCs in culture media. This non-invasive LC-MS-based analytical approach revealed significant metabolic changes between the media from control and OS-treated cells showing distinct effects of MSC differentiation on the environmental footprint of the cells in different conditions (control vs. OS treatment). A subset of compounds was directly linked to the osteogenic time-course of differentiation, and represent interesting metabolite candidates as non-invasive biomarkers for characterising the differentiation of MSCs in a culture medium.

Bio-oil and bio-char from low temperature pyrolysis of spent grains using activated alumina.

The pyrolysis of wheat and barley spent grains resulting from bio-ethanol and beer production respectively was investigated at temperatures between 460 and 540 °C using an activated alumina bed. The results showed that the bio-oil yield and quality depend principally on the applied temperature where pyrolysis at 460 °C leaves a bio-oil with lower nitrogen content in comparison with the original spent grains and low oxygen content. The viscosity profile of the spent grains indicated that activated alumina could promote liquefaction and prevent charring of the structure between 400 and 460 °C. The biochar contains about 10-12% of original carbon and 13-20% of starting nitrogen resulting very attractive as a soil amendment and for carbon sequestration. Overall, value can be added to the spent grains opening a new market in bio-fuel production without the needs of external energy. The bio-oil from spent grains could meet about 9% of the renewable obligation in the UK.

Rapid analysis of formic acid, acetic acid, and furfural in pretreated wheat straw hydrolysates and ethanol in a bioethanol fermentation using atmospheric pressure chemical ionisation mass spectrometry.

Atmospheric pressure chemical ionisation mass spectrometry (APCI-MS) offers advantages as a rapid analytical technique for the quantification of three biomass degradation products (acetic acid, formic acid and furfural) within pretreated wheat straw hydrolysates and the analysis of ethanol during fermentation. The data we obtained using APCI-MS correlated significantly with high-performance liquid chromatography analysis whilst offering the analyst minimal sample preparation and faster sample throughput.

Effect of fat content on flavor delivery during consumption: an in vivo model.

Data from studies of the effect of fat on in vivo flavor release were modeled to generate a predictive model (R(2) = 0.71). The data included a range of values from the literature and 200 new data points giving a total data set of 345 values; of these, 310 values were used as a data set for model development, and the remaining 35 values were used as a test set for model validation. The model could be used to estimate the differences in flavor delivery for samples with two different fat contents. The hydrophobicity of the flavor compounds was represented in the model by including log P. The model may provide a tool to aid in flavor reformulation between samples with different fat contents. Sensory analysis showed that an orange flavor present in a high-fat food could be reformulated for a low-fat food, giving a more similar flavor experience than in the absence of any formulation changes.

Differences in the effect of bolus weight on flavor release into the breath between low-fat and high-fat products.

The maximum intensity of flavor release increased as the weight of food introduced into the mouth (the bolus) was increased for a range of different foods. The relationship was not directly proportional (1:1) but followed a power law function. Low-fat (< or = 1 g/100 g) foods showed a different relationship than high-fat (> or = 5 g/100 g) foods, but all low-fat and all high-fat foods were broadly similar irrespective of food type or flavor molecule chemistry. For low-fat foods the intensity of flavor release increased with increasing bolus weight to a greater extent than high-fat foods. This may be associated with the capacity of fat to selectively adhere to the surfaces of the oral cavity, thereby changing the effective surface area for the release of lipophilic flavors.

Temporal aroma delivery from milk systems containing 0-5% added fat, observed by free choice profiling, time intensity, and atmospheric pressure chemical ionization-mass spectrometry techniques.

A temporal aroma delivery from milk systems containing 0, 0.5, or 5% added fat and flavored with seven-component strawberry flavoring and linalool was observed by free choice profiling (FCP), time intensity (TI), and atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) techniques. A suppressing effect of fat on the volatility of the relatively nonpolar compound linalool was observed by all methods, but only slight evidence (with the TI method) of the effect of fat on the overall strawberry (based on more polar compounds) intensity was found. With the TI method, the strawberry aroma of the fattiest sample lingered the longest, but no temporal differences were found in the release of linalool. The APCI-MS results showed no effect of fat on the temporal release of ethyl butyrate (mainly responsible for the strawberry note), but linalool of the sample containing 5% fat was found to be the most persistent. However, the effect on linalool was observed using a slightly different sampling technique than in the TI. Overall, FCP, TI, and APCI-MS showed parallel results for the effect of fat on the intensity of aroma, but temporal release data only partly supported the theory that fat slows down the release of aroma compounds and their perception.

LuxS: its role in central metabolism and the in vitro synthesis of 4-hydroxy-5-methyl-3(2H)-furanone.

Many bacteria produce extracellular molecules which function in cell-to-cell communication. One of these molecules, autoinducer 2 (AI-2), was first described as an extracellular signal produced by Vibrio harveyi to control luciferase expression. Subsequently, a number of bacteria have been shown to possess AI-2 activity in their culture supernatants, and bear the luxS gene product, which is required for AI-2 synthesis. In Porphyromonas gingivalis, luxS and pfs, encoding a 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTA/SAH'ase), form an operon, suggesting that S-adenosylhomocysteine (SAH) or 5'-methylthioadenosine (MTA) serves as a substrate for AI-2 production. Cell-free extracts of Escherichia coli MG1655, but not DH5alpha (which carries a luxS frame-shift mutation) were capable of generating AI-2 activity upon addition of SAH, but not MTA. S-Ribosyl-homocysteine (RH) derived from SAH also served as a substrate in E. coli MG1655 extracts. RH-supplemented cell-free extracts of Pseudomonas aeruginosa, a bacterium that lacks luxS, only generated AI-2 activity following the introduction of a plasmid containing the Por. gingivalis pfs-luxS operon. In addition, defined in vitro systems consisting of the purified LuxS proteins from Por. gingivalis, E. coli, Neisseria meningitidis or Staphylococcus aureus converted RH to homocysteine and a compound that exhibits AI-2 activity.4-Hydroxy-5-methyl-3(2H)-furanone was identified by mass spectrometry analysis as a major product formed in this in vitro reaction. In E. coli MG1655, expression of T3SH [the bacteriophage T3 S-adenosylmethionine (SAM) hydrolase] significantly reduced AI-2 activity in culture supernatants, suggesting that AI-2 production is limited by the amount of SAH produced in SAM-dependent transmethylase reactions. The authors suggest that the LuxS protein has an important metabolic function in the recycling of SAH. They also show that Ps. aeruginosa is capable of removing AI-2 activity, implying that this molecule may act as a nutrient. In many bacteria AI-2 may in fact represent not a signal molecule but a metabolite which is released early and metabolized in the later stages of growth.

Retronasal transport of aroma compounds.

A comparison was made between the amounts of volatiles in the headspace above a solution and the breath volatile content (exhaled from the nose or mouth) after consumption of the same solution. The amounts of volatiles in the breath were lower than those in the headspace, with breath exhaled via the mouth containing, on average, 8-fold more volatiles than breath exhaled via the nose. Dilution of the sample by saliva in-mouth did not appear to be a major factor affecting volatile delivery. Instead, the rate of in vivo equilibration (mass transfer) appeared to be the most significant factor, principally affecting volatile delivery from the solution to the gas phase. Thereafter, gas-phase dilution of the volatile as it passed through the upper airway resulted in a further decrease in volatile concentration. The final factor affecting the volatile concentration exhaled from the nose was absorption of volatiles to the nasal epithelia, which was greatest for those compounds with the lowest air/water partition coefficients.