Acute supplementation with grapes in obese subjects did not affect postprandial metabolism: a randomized, double-blind, crossover clinical trial.

PURPOSE: The aim of this study was to determine whether grape polyphenols have a "second-meal effect", modulating glucose and lipid elevations in the postprandial period after two successive meals in subjects with obesity. METHODS: A randomized, double-blind, placebo-controlled, acute clinical trial was conducted. Twenty-five obese subjects (BMI = ≥ 30 and < 40 kg/m2) were randomly divided into two groups. At an initial visit, blood was collected in a fasting state and the subjects received breakfast and 46 g of either grape powder (equivalent to 252 g fresh grapes) or placebo, both solved in water. Lunch was provided 5 h later and then blood was collected after 0, 30, 60, 120, 180, 240, 300, 330, 360, and 420 min since arrival. Two weeks later, at a second visit, the subjects received the other powder. The following were determined: glucose, insulin, triglycerides, uric acid, blood count, hemoglobin, viscosity, antioxidant capacity, and satiety perception. RESULTS: Postprandial increases were observed as expected in, for example, glucose and triglycerides after breakfast and lunch. The grape powder supplementation did not cause any significant modification compared to placebo, in these parameters; nor did it significantly modify plasma antioxidant capacity in the 6 h postprandial period. DISCUSSION: Single grape powder supplementation did not modify postprandial responses in obese subjects, probably because the polyphenol dose was insufficient to induce such an effect. The result of a combination of grape with other polyphenol-rich products or chronic supplementation with grape powder on postprandial responses remains to be elucidated. TRIAL REGISTRATION NUMBER: www.clinicaltrials.gov , NCT03741218.

Determination of volatile compounds in cider apple juices using a covalently bonded ionic liquid coating as the stationary phase in gas chromatography.

A chromatographic method for the separation of volatile compounds in Asturian cider apple juices has been developed. For this separation purpose, a monocationic imidazolium-based ionic liquid bearing a reactive terminal iodine atom was synthesized by a quaternization-anion exchange chemical sequence. Next, the gas chromatography (GC) stationary phase was prepared by covalently linking the imidazolium monolith to the reactive silanol groups of the inner capillary wall at 70 °C. This coated GC column exhibited good thermal stability (290 °C), as well as good efficiency (2000 plates/m) in the separation of volatile compounds from Asturian apple cider juices, and was characterized using the Abraham solvation parameter model. The intra-day and inter-day precision of the chromatographic method was evaluated, obtaining relative standard deviations from 3.7 to 12.9% and from 7.4 to 18.0%, respectively. Furthermore, recoveries from 82.5 to 122% were achieved. Graphical Abstract Covalent bonding of an ionic liquid to inner column wall led to a great improvement of the separation efficiencies of stationary phases in gas chromatography.

A chemometric approach to characterization of ionic liquids for gas chromatography.

A chemometric study was carried out to characterize three ionic liquid types (ILs) with hexacationic imidazolium, polymeric imidazolium, and phosphonium cationic cores, using a range of contra-anions such as halogens, thiocyanate, boron anions, triflate, and bistriflimide. The solvation parameter model developed by Abraham et al., unsupervised techniques as cluster analysis (CA), and supervised techniques as linear discriminant analysis (LDA), step-LDA, quadratic discriminant analysis (QDA), and multivariate regression techniques as discriminant partial least squares (D-PLS), or multiple linear regression (MLR) were used to characterize the functionalized ILs above. CA established two main groups of phases, those with an acidic H-bond and those with basic ones. Once detected, the two natural groups, a linear and quadratic delimiters with good classification (>96 %) and prediction (>92 %) capacities were computed. The use of step-LDA technique allowed us to establish that a, b, and s solvation parameters were the most discriminant variables. These variables were used for modeling purposes, and a D-PLS and MLR models were constructed using a binary response. The explained variance of categorical variable by the model validated by cross-validation was 65 %, and 94.5 % of ILs were correctly predicted. IL characterization carried out would allow the appropriate selection of phases for gas chromatography (GC).

Olive Pomace Oil Structuring for the Development of Healthy Puff Pastry Laminating Fats: The Effect of Chilling Storage on the Quality of Baked Products.

Developing puff pastry (PP) laminating fats (LFs) with sustainable structured olive pomace oil (OPO) could contribute to its increased valorization. This study evaluated the physicochemical stability of four OPO-based LFs or margarines and the performance of their baked PP counterparts during two months of chilling storage at 4 °C. LF samples, developed at the laboratory scale, contained 41% (LF1 and LF2) OPO and 31% (LF3 and LF4) OPO together with 10% cocoa butter when using two static initial crystallization conditions (room temperature for LF1 and LF3, freezer for LF2 and LF4) before storage. During the storage period, the proximate composition, thermal and dynamic rheological properties, firmness and spreadability, oil-binding capacity, color, and lipid oxidation of the four LF samples were examined, along with the baking performance and textural properties of the PP counterparts. The initial cooling rate had minimal significance. Cocoa butter negatively influenced post-crystallization processes occurring in OPO-based LF3 and LF4, resulting in increased hardness and reduced performance after 18 days of storage, attributed, at least partially, to a high amount of 1,3-dipalmitoyl-2-oleoyl-glycerol (POP), mainly from cocoa butter. Conversely, OPO-based LF1 and LF2 maintained their quality and were stable for two months without apparent granular crystal formation.

Enriching Eggs with Bioactive Compounds through the Inclusion of Grape Pomace in Laying Hens Diet: Effect on Internal and External Egg Quality Parameters.

(1) Background: Grapes and their associated by-products (such as grape pomace, GP) stand out for their polyphenol content, which makes them a source of bioactive compounds with antioxidant capacity. The aim of this research was to determine if the inclusion of 50 g/kg of GP in the diet of hens could enrich eggs with antioxidants and to study its effect on internal and external egg quality parameters. (2) Methods: A trial was conducted with two genetic lines of hens, which were fed either a control diet or a diet containing 50 g/kg of GP. Performance, internal and external egg quality, and egg yolk content of vitamins E and A and gallic acid were determined. (3) Results: In eggs laid by hens fed a GP diet, Haugh units and yolk color scores were enhanced, and eggshells became thinner, but without affecting the breaking strength. No dietary effect was observed on the vitamin contents of the yolk. A higher gallic acid content was observed in the yolks of eggs laid by hens fed the GP diet, suggesting that some dietary phenolic compounds could be transferred to the eggs. Hen genetics influenced egg weight, albumen Haugh units, shell thickness, and α- and γ-tocopherol concentration in yolks. (4) Conclusions: Dietary inclusion of GP improved the internal quality of eggs, enriching yolks with a phenolic compound but reducing shell thickness.

Functionality of Puff Pastry Olive Pomace Oil-Based Margarines and Their Baking Performance.

Designing healthier lipids is a current approach to developing potential functional foods. Olive pomace oil (OPO) has beneficial effects on human health, attributed to its high oleic acid content and unique bioactive compounds. Four puff pastry margarines (PP-M), based on OPO (M1, M2 at 40.8%, and M3, M4 at 30.8%, and cocoa butter at 10%) combined with low molecular weight organogelators, were prepared using two initial cooling rates (M1, M3 at 0.144 °C/min and M2, M4 at 0.380 °C/min) and compared to both commercial puff pastry (PP) butter (CB) and fatty preparation (CFP). Subsequently, six baked PP counterparts were elaborated. Physical-chemical, mechanical properties, and lipid profiles were analyzed in M1-M4 and PP, while thermal properties were determined in M1-M4. Sensory analysis was carried out in PP-M1 and PP-M3 counterparts. Elasticity (G') of M1-M4 samples was between that of controls CB and CFP, although a higher OPO content reduced viscous modulus (G″). The initial cooling rate did not affect the melting behavior of M1-M4. The firmness of PP-M1 was similar to that of PP-CB and PP-CFP, and the better spreadability and plasticity of M1 positively favored PP puffing. In addition, PP-M1 had 36.8% less SFA content than baked PP-CB, and its overall acceptability was similar. For the first time, a new margarine with high OPO content, showing adequate firmness, spreadability, and plasticity, was formulated, which gave rise to PP with appropriate performance and sensory quality and a healthy lipid profile.

Impact of Silicon Addition on the Development of Gelled Pork Lard Emulsions with Controlled Lipid Digestibility for Application as Fat Replacers.

Pork lard gelled emulsions stabilized with two proteins [soy protein concentrate (SPC) or a pork rind protein extract (PRP)], both with and without added silicon (Si) from diatomaceous earth powder, were gelled by microbial transglutaminase and к-carrageenan. These gelled emulsions (GEs), intended as fat replacers, were evaluated in different aspects, including microstructure and technological properties during chilling storage. In addition, in vitro gastrointestinal digestion (GID) with an analysis of lipolysis and lipid digestibility was also evaluated. All GEs showed adequate technological properties after 28 days of chilling storage, although the SPC-stabilized GEs showed better gravitational and thermal stability (~4% and ~6%, respectively) during chilling storage than the PRP-stabilized ones (~8 and ~12%, respectively). PRP developed larger flocculates restricting pancreatic lipase-mediated lipolysis during intestinal digestion. The addition of Si to both GE structures protected them against disruption during in vitro digestion. Accordingly, Si appears to slow down fat digestion, as reflected by higher triacylglycerides content after GID (15 and 22% vs. 10 and 18% in GEs without Si) and could become a potential candidate for use in the development of healthier meat products.

Influence of the Oil Structuring System on Lipid Hydrolysis and Bioaccessibility of Healthy Fatty Acids and Curcumin.

Oleogels (OG) and gelled emulsions (GE) were elaborated with a mixture of olive and chia oils (80:20 ratio) without and with the incorporation of the health-related compound curcumin. These were studied to evaluate the influence of the oil structuring system on the lipid hydrolysis and bioaccessibility of three healthy fatty acids (FA) (palmitic, oleic, and α-linolenic acids) and of curcumin, compared to the oil mixture (bulk oil, BO). The oil structuring system influenced the firmness and texture, and the presence of curcumin significantly altered the color parameters. GE showed higher lipid digestibility, with a greater proportion of absorbable fraction (higher content of free FA and monoacylglycerides) than OG, which behaved similarly to BO. The presence of curcumin affected the degree of lipolysis, reducing lipid digestibility in OG and increasing it in GE. As for FA bioaccessibility, although GE presented higher percentages overall, curcumin significantly increased and decreased FA bioaccessibility in OG and GE, respectively. The oil structuring system also influenced the bioaccessibility of curcumin, which was higher in GE. Therefore, when selecting an oil structuring system, their physicochemical properties, the degree of lipid hydrolysis, and the bioaccessibility of both curcumin and the FA studied should all be considered.

Determination of extractable pollutants from microplastics to vegetables: Accumulation and incorporation into the food chain.

The presence and impacts of microplastics (MPs) are being extensively researched and reviewed, especially in the marine environment. However, mobility, transportation routes, and accumulation of leaching compounds such as additives in plastic waste including MPs are scarcely studied. Information regarding ecotoxicity and leachability of compounds related to MPs contamination in the environment is limited. Current work presents the levels of leachates from plastic materials in edible-root and non-edible root vegetables. Samples were analyzed by static headspace and gas chromatography-mass spectrometry (SHS-GC-MS) and the presence of 93 putative compounds was accurately monitored in the samples by the usage of Mass Spectrometry-Data Independent Analysis software. The application of chemometrics to the SHS-GC-MS dataset allowed differentiation between the levels of plastic related compounds in edible root and non-edible root vegetables, the former showing a higher content of plastic leachates. For SHS sampling, 3 g of the sample were incubated at 130 °C for 35 min in the HS vial and toluene and naphthalene were added as internal standards for quantification purposes. The developed SHS-GC-MS methodology is straightforward, reliable, and robust and allowed the quantification of sixteen plastic associated compounds in the samples studied in a range from 0.14 to 28800 ng g-1 corresponding to 2,4-di-tert-butylphenol and p,α-dimethylstyrene, respectively. Several of the quantified compounds pointed out to potential contamination of polystyrene and/or polyvinyl chloride MPs.

Development and Physico-Chemical Characterization of Healthy Puff Pastry Margarines Made from Olive-Pomace Oil.

Due to its characteristic aroma and flavor, puff pastry (PP) prepared with butter is more accepted than that made with margarine, yet a high saturated fat consumption is associated with cardiovascular disease. This work studies the potential of olive-pomace oil (OPO) as main ingredient of PP margarines together with different organogelator agents to imitate the technological properties of a commercial fatty preparation (CFP). Rheological and textural properties were measured in all formulated margarines (FM) and, additionally, thermal and microstructural properties, and fatty acid (FA) profiles were analyzed in some selected FM. The different FM had viscous modulus (G″) and loss factor (tan δ) values lower than those of CFP, thus reflecting a different viscoelasticity and plasticity. The crystallization and melting temperatures of FM were also different from those of CFP, indicating the presence of a dissimilar polymorphic fat-crystal structure. Nevertheless, the FM containing an oleogel prepared with 5% beeswax and OPO was more similar to CFP. The FA profile of CFP and FM, with 80% polyunsaturated fatty acids (PUFA) and 60% oleic acid, is healthier than that of a PP commercial butter (CB), evidencing that, although improvements in margarine plasticity are still necessary, OPO is technologically viable to produce healthier PP margarines.

The Influence of Cellulose Ethers on the Physico-Chemical Properties, Structure and Lipid Digestibility of Animal Fat Emulsions Stabilized by Soy Protein.

This study explores the influence of carboxymethylcelullose (CMC) and methylcelullose (MC), added by simultaneous (sim) and sequential (seq) emulsification methods, on the structure, rheological parameters and in vitro lipid digestibility of pork lard O/W emulsions stabilized by soy protein concentrate (SPC). Five emulsions (SPC, SPC/CMC-sim, SPC/CMC-seq SPC/MC-sim, SPC/MC-seq) were prepared in vitro. The presence of CMC and MC, and the stage of incorporation affected the emulsion microstructure. In the SPC emulsion, lipid droplets were entrapped by a protein layer that was thicker when MC was added, providing greater resistance against environmental stresses during gastrointestinal digestion. At 37 °C, CMC incorporation produced a structural reinforcement of the SPC emulsion, whereas MC addition did not affect the network rigidity, although a delaying effect on the crossover temperature was observed, which was more evident in SPC/MC-seq. The presence and stage of CMC and MC incorporation affected the rate and extent of lipolysis, with SPC/MC-seq presenting an inferior concentration of free fatty acids. The lower extent of lipolysis observed in SPC/MC-seq may be positive in the manufacture of animal fat products in which reduced fatty acid absorption is intended.

Insights on the effect of age and gender on in-mouth volatile release during wine tasting.

This study focus for the first time, in looking for age-gender effects on in vivo volatile release during wine consumption, also considering oral physiological differences (e.g. saliva composition). To do so, the in-mouth Head Space Sorptive Extraction technique was used, which allowed monitoring the oral release of twenty-four different types of volatile compounds from white and red wines. Thirty-two individuals (n = 32) males and females, belonging to two different age groups: young (18-35 y.o) and senior (>55 y.o.) participated in this analytical in vivo study. Results showed differences in volatile release among age-gender groups, which also depended on the volatile compound and wine type. Senior groups (SM, SF) showed a similar release behaviour among them. Contrarily, young males showed a higher release (between 10 and 29%) of alcohols and esters indistinctly of the wine type, while young females showed the lowest oral volatile release among the four age-gender groups. Gender differences in volatile release were more evident in young than in seniors. A higher release of furanic compounds (furfural and 5-methyl furfural) in seniors was likely related to differences in their saliva composition (total protein content, minerals (Mg, Zn) and α-amylase activity).

Transcriptomic analysis of CO2-treated strawberries (Fragaria vesca) with enhanced resistance to softening and oxidative stress at consumption.

One of the greatest threats to wild strawberries (Fragaria vesca Mara des Bois) after harvest is the highly perishability at ambient temperature. Breeders have successfully met the quality demands of consumers, but the prevention of waste after harvest in fleshy fruits is still pending. Most of the waste is due to the accelerated progress of senescence-like process after harvest linked to a rapid loss of water and firmness at ambient temperature. The storage life of strawberries increases at low temperature, but their quality is limited by the loss of cell structure. The application of high CO2 concentrations increased firmness during cold storage. However, the key genes related to resistance to softening and cell wall disassembly following transference from cold storage at 20°C remain unclear. Therefore, we performed RNA-seq analysis, constructing a weighted gene co-expression network analysis (WGCNA) to identify which molecular determinants play a role in cell wall integrity, using strawberries with contrasting storage conditions, CO2-cold stored (CCS), air-cold stored (ACS), non-cold stored (NCS) kept at ambient temperature, and intact fruit at harvest (AH). The hub genes associated with the cell wall structural architecture of firmer CO2-treated strawberries revealed xyloglucans stabilization attributed mainly to a down-regulation of Csl E1, XTH 15, Exp-like B1 and the maintenance of expression levels of nucleotide sugars transferases such as GMP and FUT as well as improved lamella integrity linked to a down-regulation of RG-lyase, PL-like and PME. The preservation of cell wall elasticity together with the up-regulation of LEA, EXPA4, and MATE, required to maintain cell turgor, is the mechanisms controlled by high CO2. In stressed air-cold stored strawberries, in addition to an acute softening, there is a preferential transcript accumulation of genes involved in lignin and raffinose pathways. Non-cold stored strawberries kept at 20°C after harvest are characterized by an enrichment in genes mainly involved in oxidative stress and up-expression of genes involved in jasmonate biosynthesis. The present results on transcriptomic analysis of CO2-treated strawberries with enhanced resistance to softening and oxidative stress at consumption will help to improve breeding strategies of both wild and cultivated strawberries.

Evaluation of new ionic liquids as high stability selective stationary phases in gas chromatography.

Two ionic liquids (ILs), namely (S,S)-1-butyl-3-(2'-hydroxy-cyclohexyl)-3H-imidazol-1-ium tetrafluoroborate and (S,S)-1-butyl-3-(2'-acetyl-cyclohexyl)-3H-imidazol-1-ium tetrafluoroborate have been employed as stationary phases in capillary gas chromatography. These new phases exhibit a column efficiency of 1,600 and 2,100 plates m(-1) for IL 1 and IL 2, respectively, a wide operating temperature range and good thermal stability (bleeding temperature of 250 °C for IL 1 and 160 °C for IL 2). Inverse gas chromatography (GC) analyses were used to study the solvation properties of these ILs through a linear solvation energy model. The application of these ILs as new GC stationary phases was studied. These stationary phases exhibited unique selectivity for many organic substances, such as alkanes, ketones, esters, and aromatic compounds. The efficient separation of several mixtures containing compounds of different polarities and the good separation of fatty acid methyl esters (FAMEs) and cis/trans isomers indicate that these ILs may be applicable as a new type of GC stationary phases.

Experimental design applied to the analysis of volatile compounds in apple juice by headspace solid-phase microextraction.

A simple and fast method based on solid-phase microextraction (SPME) followed by fast gas chromatography (Fast GC) has been developed for the analysis of volatile compounds in Asturian apple juices employed in the cider production. Three different fiber coatings have been checked (PDMS, PDMS-DVB and CAR-PDMS) and PDMS-DVB has been presented to be the most suitable one. Experimental design has been employed in the optimization of extraction factors and robustness assessment. The use of Fast GC allowed the separation of 14 compounds (esters, aldehydes and alcohols) in approximately 4 min, clearly reducing the analysis time when compared to conventional GC. Good linearity, recoveries and repeatability of the solid-phase microextraction were obtained with r(2) values, recoveries and relative standard deviations ranging from 0.9822 to 0.9998, 83.2 to 109.8% and 0.5 to 11.7%, respectively, using standard solution.

Development of Chocolates with Improved Lipid Profile by Replacing Cocoa Butter with an Oleogel.

The reformulation of chocolates seeks to find innovative alternatives to cocoa butter (CB) that are more economical and adhere to nutritional recommendations to replace saturated fats with unsaturated ones. In this research, chocolates were elaborated by substituting CB with an oleogel (OG) formulated with hydroxypropyl methylcellulose (HPMC) as an entrapper of sunflower oil by using the foam-templated approach. Four different CB/OG blends were prepared and characterized as potential CB substitutes (100/0 control), at replacement levels of 30%, 50%, 70% and 100% (70/30, 50/50, 30/70 and 0/100 blends), and subsequently, CB/OG-based chocolates (CB/OG-Ch) were formulated (100/0-Ch, 70/30-Ch, 50/50-Ch, 30/70-Ch and 0/100-Ch). Both the CB/OG blends and the CB/OG-Ch counterparts were characterized by dynamic and stationary rheology, hardness, thermal parameters, microstructure, and oil-binding capacity; in addition, the lipid profile of the chocolates was analyzed, and a sensory analysis was performed. Increasing the OG proportion in the CB/OG blend weakens the rigidity and strength of the fat-crystal network conferred by the CB, and decreases both its viscoelasticity and thermal parameters, but the differences between all the different properties and parameters of the CB/OG-Ch samples diminished in presence of the other ingredients used in the chocolate formulation. Sensory analysis evidenced that it is possible to replace up to 70% of CB with the OG, although from a technological point of view a replacement level of 50% would seem more appropriate. As compared to 100/0-Ch, 50/50-Ch and 30/70-Ch involve saturated fat reductions of 55% and 37%, respectively.

Thermorheological Characterization of Healthier Reduced-Fat Cocoa Butter Formulated by Substitution with a Hydroxypropyl Methylcellulose (HPMC)-Based Oleogel.

Cocoa butter (CB) is a main ingredient in pastry due to the unique functional properties of its fat, which is high in saturated fatty acids (SFAs). However, excessive consumption of SFAs is associated with the occurrence of several chronic diseases. This study researched the partial or total replacement of CB by an oleogel (OG) formulated with a healthier lipid profile, for mixed systems that would allow a partial substitution of CB in confectionery products. The "emulsion-templated approach" was used to develop a sunflower oil-HPMC-based OG. Different CB:OG ratios were formulated increasing the percentage replacement of CB by OG from 50 to 100%. Rheological and textural properties were determined and compared with a CB control at 20 and 10 °C. Oil-binding capacity was also analyzed. The systems showed a solid-like behavior, with higher elastic than viscous modulus, which increased with CB concentration. Compared with 20 °C, at 10 °C there was an increase in connectivity, viscoelasticity, and consistency of the systems, in response to a more complete CB crystallization. The replaced systems also presented a better lipid profile than CB. This evidence suggests that formulated CB:OG system at 50:50 ratio could become useful as a CB equivalent in chocolate products.

Foods for Plant-Based Diets: Challenges and Innovations.

Plant-based diets have become popular as a means of reducing the environmental footprint of the diet and promoting human health and animal welfare. Although the percentages of vegetarians and vegans are low compared to omnivores, their numbers have increased significantly in the last years. The use of non-animal food products other than meat alternatives is also increasing and this tendency constitutes an opportunity for the food industry. In this review, we present that plant-based meat and milk alternatives are consolidated but that there is a niche for egg, seafood alternatives, and new products which may not resemble any traditional animal food. However, not all animal food substitutes are sustainable and some of them are even ultra-processed. In addition, there are concerns on safety and labeling, and consumers demand clear information and regulation. The challenges in this field are connected with food design and technology, sensory science, nutrition, and dietetics. Moreover, adequate selection and combination of foods is important in order to achieve consumer acceptance while preventing nutritional deficiencies in those who choose this type of diet.

Assessment of the Miniature Kramer Shear Cell to Measure Both Solid Food and Bolus Mechanical Properties and Their Interplay with Oral Processing Behavior.

This study assessed the usefulness of the miniature Kramer shear cell to determine relevant instrumental parameters of solid foods and bolus counterparts, examining their relationships with oral processing behaviors to obtain greater knowledge about the texture perception process. Six solid foods with different textural properties were tested. Bolus mechanical properties were also determined by means of cone penetration tests and rheological measurements, and their particle size distributions by sieving. Oral processing behavior (chewing time, number of chews, chewing rate, eating rate) and food saliva uptake (SU) of a young volunteer and a panel of 39 untrained participants were analyzed. The Kramer mechanical properties were very suitable for detecting different levels of food and bolus textural hardness and fracturability and the associated degrees of fragmentation achieved during mastication. Chewing time and number of chews were highly correlated with Kramer food and bolus mechanical properties for the single subject and for the panel's oral processing behaviors. For the single subject, SU and eating rate also showed strong correlations with food and bolus mechanical properties, unlike chewing rate and food moisture content (FMC). In contrast, eating rate, FMC, and SU did not vary with the oral activities of the panel.

Understanding the crispy-crunchy texture of raw red pepper and its change with storage time.

Red sweet peppers held in cold storage were periodically sampled at 1-week intervals over a 3-weeks period using three-point bending, puncture, cutting, and Volodkevich (coupled with acoustic emission) tests, confocal laser scanning microscopy (CLSM) and other physicochemical measurements. At each sampling, tissue specimens were soaked in mannitol solutions (0.0-0.9M) and puncture test, dimension changes and CLSM were used to identify degrees of turgidity present in osmotically manipulated pepper tissue. Pepper texture became crumbly with increased storage time due to softening and wilting processes. The Young's modulus, derived from the bending test using the single-edge notched bend geometry without notches decreased progressively during cold storage and resulted as the best mechanical parameter for measuring the loss of whole-tissue stiffness by both decreased cell wall stiffness and turgor pressure. Osmotic adjustment indicated that the pepper structure is extremely anisotropic, with the specimen's "average" relative thickness (RT) being the dimension change more affected. Incipient plasmolysis was evident in the highest mannitol concentration (0.9M), therefore, the turgor pressure of nonsoaked tissue could not be inferred. However, significant correlations were found between RT and puncture parameters such as initial slope, initial and final distances, and the number of flesh and skin force peaks, which depended on the dilation or shrinkage caused by the osmotic adjustment. During storage, soaked tissues had lower crunchy texture than nonsoaked, reflecting that cell wall stiffness plays a more significant role in determining pepper crunchiness than cell turgor pressure.