A lubrication analysis of pharyngeal peristalsis: Application to flavour release

After eating a liquid or a semi-liquid food product, a thin film responsible for the dynamic profile of aroma release coats the pharyngeal mucosa. The aim of this article was to analyse the fluid mechanics of pharyngeal peristalsis and to develop a simple biomechanical model in order to understand the role of saliva and food bolus viscosity on the coating of pharyngeal mucosa. We began by analysing the physiology and the biomechanics of swallowing in order to determine relevant model assumptions. This analysis of the literature clarified the types of mechanical solicitations applied on the food bolus. Moreover, we showed that the pharyngeal peristalsis in the most occluded region is equivalent to a forward roll coating process, the originality of which is lubrication by a film of saliva. A model based on the lubrication theory for Newtonian liquids was developed in dimensionless form. The parametric study showed the strong influence of relative saliva thickness on the food bolus coating. A specific experimental device was designed that confirms the model predictions. Two sets of conditions that depend on the relative thickness of saliva were distinguished. The first is characterised by a relatively thin film of saliva: food bolus viscosity has a strong impact on mucosa coating. These phenomena are well represented by the model developed here. The second is obtained when the saliva film is relatively thick: hydrodynamic mixing with saliva, interdiffusion or instabilities may govern mucosa coating. Finally, these results were extrapolated to determine the influence of food bolus viscosity on the dynamic profile of flavour release according to physiological parameters.     

Flavor release of diacetyl and 2-heptanone from cream style dressings in three mouth model systems

The release of volatile compounds from a cream style dressing, which consisted of a thickening agent dispersed in the water phase of an oil in water (o/w) type of emulsion, was studied by the purge-and-trap (PT), dynamic head space mastication (DHM) and dynamic headspace (DH) model systems for diacetyl and 2-heptanone as two volatile compounds. Big differences were detected in the quantity of volatiles released by the three models for both diacetyl and 2-heptanone: PT released the most, followed by DHM and DH. Nitrogen gas bubbling in PT and plunger up-and-down motion in DHM mimic mouth movements and promoted volatile release more than DH. The quantity of volatiles released depended on the nitrogen gas flow rate and isolation period with both the PT and the DHM model. Static headspace measurements indicated that no interaction occurred between the volatiles and the dispersion thickening agent, nor between the volatiles and protein of saliva.     

Volatile release from liquids: a comparison of in vivo APCI-MS, in-mouth headspace trapping and in vitro mouth model data

In-mouth volatile release from flavoured water was followed using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) or using a hand-held, computer-controlled device based on sequential trapping of flavours on Tenax traps. The present results verify recent in vitro data obtained with a sophisticated, fully computerized mouth model apparatus and confirm its validity for the simulation of in-mouth dynamic volatile release. In-nose APCI-MS measurements showed considerable person-to-person variability in non-trained individuals during drinking due to subconscious control of muscles during swallowing and subsequent breathing. Data showed a 'swallow breath' volume reaching the nasal cavity from the throat, not from the mouth cavity. Flavour enriched air from the mouth was shown to be transported to the nose (via exhalation) immediately after the swallowing event, but the dynamic process of volatile equilibration between residuals of the swallowed liquid and the exhaled air predominantly determined volatile in-nose concentration. Owing to its dynamic character, the process of volatile equilibration and release in the throat upon exhalation should be similar to the in-mouth process studied in the present work. A full mechanical simulation of retronasal volatile transport, however, will remain difficult.     

A biomechanical model of swallowing for understanding the influence of saliva and food bolus viscosity on flavor release

After swallowing a liquid or a semi-liquid food product, a thin film responsible for the dynamic profile of aroma release coats the pharyngeal mucosa. The objective of the present article was to understand and quantify physical mechanisms explaining pharyngeal mucosa coating. An elastohydrodynamic model of swallowing was developed for Newtonian liquids that focused on the most occluded region of the pharyngeal peristaltic wave. The model took lubrication by saliva film and mucosa deformability into account. Food bolus flow rate and generated load were predicted as functions of three dimensionless variables: the dimensionless saliva flow rate, the viscosity ratio between saliva and the food bolus, and the elasticity number. Considering physiological conditions, the results were applied to predict aroma release kinetics.Two sets of conditions were distinguished. The first one was obtained when the saliva film is thin, in which case food bolus viscosity has a strong impact on mucosa coating and on flavor release. More importantly, we demonstrated the existence of a second set of conditions. It was obtained when the saliva film is thick and the food bolus coating the mucosa is very diluted by saliva during the swallowing process and the impact of its viscosity on flavor release is weak. This last phenomenon explains physically in vivo observations for Newtonian food products found in the literature. Moreover, in this case, the predicted thickness of the mix of food bolus with saliva coating the mucosa is approximately of 20 μm; value in agreement with orders of magnitude found in the literature.     

Sensory perception is related to the rate of change of volatile concentration in-nose during eating of model gels.

The relationship between perceived aroma and the volatile concentration measured in-nose was investigated during eating of a model food. Sensory ranking and time-intensity analysis (TI) were used to measure perceived aroma, while in-nose volatile concentration was monitored by atmospheric pressure ionization mass spectrometry, which produced time release data. A gelatine-sucrose gel with a range of gelatine concentrations (2-8% w/w) and flavoured with furfuryl acetate was used as the model food. Sensory scaling showed decreased flavour intensities and TI showed a decrease in the flavour perceived over time, as the gelatine concentration increased. Studies in model systems and in people demonstrated that the different rates of release observed for different gelatine concentrations were not due to binding of volatile to protein in the gel, nor to mucous membranes, but were due to different rates of gel breakdown in-mouth. There were no significant differences in the maximum in-nose volatile concentrations for the different gelatine concentrations, so the amount of volatile present did not correlate well with the sensory analysis. However, the rates of volatile release were different for the different gels and showed a good correlation with sensory data.     

Release cells, breath analysis and in-mouth analysis in flavour research

The flavour of a food or beverage is not perceived in a single event, but rather as a series of events experienced as the food is consumed. Recent methods in flavour research have taken account of this, and techniques have been developed to study flavour release in model systems (release cells or simulated mouths) and from the mouth or nose of assessors, while consuming foods. However, while there is agreement on the need in some cases for hydration or artificial saliva in simulated mouths, other parameters must be optimised on a case-by-case basis. Individual variability may still be a problem in breath analysis, and further work is required to determine the extent to which there are real differences in volatile profiles. The techniques of release cells and breath analysis must now be applied to provide data, which will allow flavour release to be modelled.     

Modeling the kinetics of flavour release during drinking

Novel mathematical models for flavour release during drinking are described, based on the physiology of breathing and swallowing. Surprisingly, we conclude that most flavour molecules arriving in the nose are extracted from liquid left in the throat, after swallowing. The models are fit to real time flavour release data obtained using APCI-mass spectrometry. Before modelling, raw data are corrected for the effects of varying airflow rate, using the signal from acetone in exhaled air. A simple equilibrium batch extraction model correctly describes flavour release during the first breaths after swallowing a flavoured liquid. It shows that for eight volatiles, whose in vitro air-water partition coefficients vary by a factor of 500, the apparent in vivo air-saliva partition coefficients vary only by a factor of five. To interpret the kinetics of flavour release longer after swallowing, diffusion of flavour into the throat lining is included. This is done using a three-layer model for mass transfer in the throat. An analytical solution of this model gives good fits to typical data. These models de-couple the physiological and physico-chemical aspects of flavour release, clarifying the effect of behaviour on in-vivo flavour release.     

Flavour retention and release from protein solutions

This paper briefly presents the main results obtained up to now on protein-flavour binding and release in relation with flavour perception. Among the food proteins, beta-lactoglobulin is the most extensively studied for its binding properties, which involve both hydrophobic and hydrogen binding. Recent developments using molecular modelling and Quantitative Structure-Activity Relationship confirmed the existence of two different binding sites for flavour compounds on beta-lactoglobulin. During the aroma release process in the mouth, not only free aroma compounds are released but also those reversibly bound by the protein, pointing out the fact that flavour perception is only affected if strong binding occurs.     

Criteria for mucus transport in the airways by two-phase gas-liquid flow mechanism

The critical conditions for mucous layer transport in the respiratory airways by two-phase gas-liquid flow mechanism were investigated by using 0.5- and 1.0-cm-ID tube models. Several test liquids with rheological properties comparable to human sputum were supplied continuously into the vertically positioned tube models in such a way that the liquid could form a uniform layer while traveling upward through the tube with a continuous upward airflow. The critical airflow rate and critical liquid layer thickness required for the upward transport of the liquids were determined. The critical airflow rate was in the Reynolds number (Re) range of 142-1,132 in the 0.5-cm-ID tube model and 708-2,830 in the 1.0-cm-ID tube model depending on the types of liquids tested. In both models, the critical airflow rate was lower with viscoelastic liquids than with viscous oils. The critical liquid layer thickness ranged from 0.2 to 0.5 mm in the 0.5-cm-ID tube model and 0.8 to 1.4 mm in the 1.0-cm-ID tube model at Re of 2,800. These values decreased rapidly with increasing airflow rate. The critical thickness relative to the tube diameter ranged from 3 to 15% of the respective tube diameter and was lower by approximately 30-50% in the 0.5-cm-ID tube model than in the 1.0-cm-ID tube model over the entire Re range tested. The results indicate that the critical conditions for the mucus transport by two-phase gas-liquid flow mechanism are within the range that can be achieved in patients with bronchial hypersecretions during normal breathing.     

Estimation of the mechanical parameters of the human respiratory system

A numerical algorithm has been developed for the estimation of the mechanical parameters of the human respiratory system. In order to estimate the pulmonary resistance and the dynamic pulmonary elastance, the transpulmonary pressure and the airflow at the mouth or nose are expanded in Chebyshev series. The nonlinear mathematical lung model and a set of measurements for airflow and pressure are then handled by the numerical technique. The lung model includes a component to account for turbulent flow in the larynx and trachea. This contribution presents an alternative method for lung parameter estimation and differs from most existing methods in that it does not need measurements for the tidal volume. It therefore eliminates the use of a body plethysmograph. The method may also find potential application to various other parameter identification problems.     

Solid-phase microextraction for determining twelve orange flavour compounds in a model beverage emulsion

Solid-phase microextraction (SPME) coupled to gas chromatography has been applied for the headspace analysis (HS) of 12 target flavour compounds in a model orange beverage emulsion. The main volatile flavour compounds studied were: acetaldehyde, ethyl acetate, alpha-pinene, ethyl butyrate, beta-pinene, myrcene, limonene, gamma-terpinene, octanal, decanal, linalool and citral (neral plus geranial). After screening the fibre type, the effect of other HS-SPME variables such as adsorption temperature (25-55 degrees C), extraction time (10-40 min), sample concentration (1-100% w/w), sample amount (5-10 g) and salt amount (0-30% w/w) were determined using a two-level fractional factorial design (2(5-2)) that was expanded further to a central composite design. It was found that an extraction process using a carboxen-polydimethylsiloxane fibre coating at 15 masculineC for 50 min with 5 g of diluted emulsion 1% (w/w) and 30% (w/w) of sodium chloride under stirring mode resulted in the highest HS extraction efficiency. For all volatile flavour compounds, the linearity values were accurate in the concentration ranges studied (r(2) > 0.97). Average recoveries that ranged from 90.3 to 124.8% showed a good accuracy for the optimised method. The relative standard deviation for six replicates of all volatile flavour compounds was found to be less than 15%. For all volatile flavour compounds, the limit of detection ranged from 0.20 to 1.69 mg/L.     

Mucus transport in the airways by two-phase gas-liquid flow mechanism: continuous flow model

Mucus transport speed induced by two-phase gas-liquid interaction was measured in the continuous two-phase annular flow tube models, and factors influencing the transport speed were assessed in conjunction with rheological properties of mucus. The flow model was made with 1.0-cm-ID glass tubes and positioned either vertically or horizontally. During a continuous passage of airflow through the model tube, mucus stimulants were supplied into the tube at a rate of 0.5-2.0 ml/min. The advancing speed of the leading edge of the mucous layer and mean mucous layer thickness were then measured. The transport speed in the vertical tube model ranged from 1.1 to 3.1 cm/min with a mucus feed rate of 0.5 ml/min at airflow rates of 0.33-1.17 l/s and increased with increasing airflow rates but decreased rapidly with increasing viscosity of mucus. The transport speed increased almost proportionally with increasing mucus feed rate. Elasticity of mucus did not affect the transport speed itself. However, more elastic mucus caused lower flow resistance and thereby could be transported with a much reduced work load. The transport speed in the horizontal tube model was 5-60% faster than that in the vertical tube model. The mean mucous layer thickness in the vertical tube model was found to be in the range of 0.5-1.5 mm in the experimental conditions used, and decreased rapidly with increasing airflow rate and decreasing viscosity of mucus. From these data the transport speed could be functionally related to airway diameter, mucous layer thickness, and mucus production rate.     

Biomechanical Aspects of Compliant Airways due to Mechanical Ventilation

Without proper knowledge of mechanical ventilation effects, physicians can aggravate an existing lung injury. A better understanding of the interaction between airflow and airway tissue during mechanical ventilation will be helpful to physicians so that they can provide appropriate ventilator parameters for intubated patients. In this study, a computational model incorporating the interactions between airflow and airway walls was developed to investigate the effects of airway tissue flexibility on airway pressure and stress. Two flow rates, 30 and 60 l/min, from mechanical ventilation were considered. The transient waveform was active inhalation with a constant flow rate and passive exhalation. Results showed that airway tissue flexibility decreased airway pressure at bifurcation sites by approximately 25.06% and 16.91% for 30 and 60 l/min, respectively, and increased wall shear stress (WSS) by approximately 74.00% and 174.91% for 30 and 60 l/min, respectively. The results from the present study suggested that it is very important to consider the interaction between airflow and airway walls when computational models are developed. Results of this study help to better quantify how the airflow rate used in mechanical ventilation, in conjunction with airway tissue flexibility, affects airway pressure and stresses.     

Salivary β-glucosidase as a direct factor influencing the occurrence of halitosis

β-Glucosidases are enzymes present in all living organisms, playing a pivotal role in diverse biological processes. These enzymes cleave β-glycosidic bonds between carbohydrates, or between a carbohydrate and a non-carbohydrate moiety, which may result in the liberation of volatile aglycones. Released compounds execute diverse physiological roles, while the industry takes advantage of exogenously added β-glucosidases for aroma enrichment during food and beverage production. β-Glucosidase enzymatic activity has been reported in human saliva and given the fact that these enzymes are involved in aroma release, we investigated here the correlation between β-glucosidase activity in human saliva and the occurrence of halitosis. Measurement of salivary enzyme activity of 48 volunteers was performed using p-nitrophenyl-β-d-glucopyranoside as substrate. Each volunteer was clinically evaluated by a dental surgeon and clinical and laboratorial data were statistically analyzed. Gas-chromatography of saliva headspace allowed the analysis of the direct role of exogenous β-glucosidase on aromatic /volatile profile of saliva samples. The data demonstrated a positive correlation between halitosis and enzymatic activity, suggesting that the enzyme exerts a direct role in the occurrence of bad breath. Gas-chromatography analysis demonstrated that exogenously added enzyme led to the alteration of volatile organic content, confirming a direct contribution of β-glucosidase activity on saliva volatile compounds release. Although halitosis is a multifactorial condition, the complete understanding of all governing factors may allow the development of more effective treatment strategies. Such studies may pave the way to the use of β-glucosidase inhibitors for halitosis clinical management.     

Computational model of particle deposition in the nasal cavity under steady and dynamic flow

A computational model for flow and particle deposition in a three-dimensional representation of the human nasal cavity is developed. Simulations of steady state and dynamic airflow during inhalation are performed at flow rates of 9–60 l/min. Depositions for particles of size 0.5–20 μm are determined and compared with experimental and simulation results from the literature in terms of deposition efficiencies. The nasal model is validated by comparison with experimental and simulation results from the literature for particle deposition under steady-state flow. The distribution of deposited particles in the nasal cavity is presented in terms of an axial deposition distribution as well as a bivariate axial deposition and particle size distribution. Simulations of dynamic airflow and particle deposition during an inhalation cycle are performed for different nasal cavity outlet pressure variations and different particle injections. The total particle deposition efficiency under dynamic flow is found to depend strongly on the dynamics of airflow as well as the type of particle injection.     

Retro-nasal aroma release is correlated with variations in the in-mouth air cavity volume after empty deglutition

We hypothesized that interindividual differences in motor activities during chewing and/or swallowing were determining factors for the transfer of volatile aroma from the in-mouth air cavity (IMAC) toward the olfactory mucosa. In our first experiment, we looked for changes in IMAC volume after saliva deglutition in 12 healthy subjects. The mean IMAC volume was measured after empty deglutition using an acoustic pharyngometer device. Based on the time course of the IMAC volume after swallowing, we discerned two groups of subjects. The first group displayed a small, constant IMAC volume (2.26 mL ±0.62) that corresponded to a high tongue position. The second group displayed a progressive increase in IMAC (from 6.82 mL ±2.37 to 22.82 mL ±3.04) that corresponded to a progressive lowering of the tongue to its resting position. In our second experiment, we investigated the relationship between IMAC volume changes after deglutition and the level of aroma release at the nostril. For this purpose, the release of menthone was measured at the nostril level in 25 subjects who consumed similar amounts of a mint tablet. The subjects were separated into two groups corresponding to two levels of menthone release: high (H) and low (L). The mean volume of IMAC was measured during and after empty deglutition. Group H displayed a small, constant amplitude of IMAC volume change after deglutition, while Group L displayed a progressive increase in IMAC. It is likely that Group H continuously released the aroma through the veloglossal isthmus as the mint was consumed, while Group L trapped the aroma in the oral cavity and then released it into the nasal cavity upon swallowing. These results show that the in vivo aroma release profile in humans depends closely on the different motor patterns at work during empty deglutition.     

Fly-attracting volatiles produced by Rhodococcus fascians and Mycobacterium aurum isolated from myiatic lesions of sheep.

Bacterial strains isolated from the healthy breech mucosa and myiatic wounds of ewes were tested for their volatile production as fly attractants towards Wohlfahrtia magnifica (Diptera: Sarcophagidae). Cultures were studied as fly baits in field experiments, and strains performing with the best chemotropic effect were selected for further analysis. Static and dynamic headspace samples from shaken cultures were examined by gas chromatography-mass spectrometry (GC-MS). Strains identified as Rhodococcus fascians and Mycobacterium aurum produced various volatile sulfur compounds and benzene, and proved to be the best fly attractants.     

Metabolite profiling of the ripening of Mangoes <Emphasis Type="Italic">Mangifera indica</Emphasis> L. cv. ‘Tommy Atkins’ by real-time measurement of volatile organic compounds

Real-time profiling of mango ripening based on proton transfer reaction-time of flight-mass spectrometry (PTR–ToF–MS) of small molecular weight volatile organic compounds (VOCs), is demonstrated using headspace measurements of ‘Tommy Atkins’ mangoes. VOC metabolites produced during the ripening process were sampled directly, which enabled simultaneous and rapid detection of a wide range of compounds. Headspace measurements of ‘Keitt’ mangoes were also conducted for comparison. A principle component analysis of the results indicated that several mass channels were not only key to the ripening process but could also be used to distinguish between mango cultivars. The identities of 22 of these channels, tentatively speciated using contemporaneous GC–MS measurements of sorbent tubes, are rationalized through examination of the biochemical pathways that produce volatile flavour components. Results are discussed with relevance to the potential of headspace analysers and electronic noses in future fruit ripening and quality studies.     

Development and validation of method for analysis of some ototoxic solvents in saliva matrix by headspace gas chromatography/mass spectrometry

The aim of this study was to develop an analytical method to monitor the saliva matrix for ototoxic solvents absorption: the method is based on headspace gas chromatography/mass spectrometry and represents an alternative biological monitoring for investigating low exposure to hazardous ototoxic solvents. Simultaneous determination of toluene, ethylbenzene, xylenes and styrene has been carried out and the method has been optimized for both instrumental parameters and samples treatment. Chromatographic conditions have been set in order to obtain a good separation of xylene isomers due to the interest in p-xylene as ototoxic one. Method validation has been performed on standards spiked in blank saliva by using two internal standards (2-fluorotoluene and deuterated styrene-d₈). This method showed the possibility to detect the target compounds with a linear dynamic range of at least a 2 orders of magnitude characterized by a linear determination coefficient (r²) greater than 0.999. The limit of detection (LOD) ranged between 0.19 ng/mL (styrene) and 0.54 ng/mL (m-xylene) and the lower limit of quantification (LLOQ) ranged between 0.64 ng/mL (styrene) and 1.8 ng/mL (m-xylene). The method achieved good accuracy (from 99 to 105%) and precision for both intra- and inter-assay (relative standard deviation ranging from 1.7 to 13.8%) for all six compounds concerned. The repeatability was improved by adding sodium sulphate to the matrix. Saliva samples resulted stable for at least 7 days after collection, if stored in headspace vials, at the temperature of 4 °C. An evaluation of the main sources of uncertainty of the method is also included: expanded uncertainties ranges between 10 and 16% for all of the target compounds. In summary, the headspace gas chromatography/mass spectrometry method is a highly sensitive, versatile and flexible technique for the biological monitoring of exposure to ototoxic solvents by saliva analysis.