Black Truffle Aroma Evaluation: SPME-GC-MS vs. Sensory Experts.

Nowadays, the truffle aroma attribute is not included as a quality parameter in the current recommendation that explains the truffle quality (UNECE standard 53 FFV3) and establishes the truffle commercial categories. However, the aroma is the main reason why truffles are worldwide appreciated. Indeed, more than 30 aromatic molecules compose it, and this is the reason why the human evaluation and identification of these odorants, without previous training, is quite subjective. Analytical techniques such as gas chromatography techniques, however, can establish an aromatic profile and detect potential aromatic markers. In this study, 16 tasting experts were trained to make more objective the truffle aroma evaluation and odorants identification. For this, a comparison between solid-phase microextraction gas chromatography coupled with mass spectrometry (SPME-GC-MS) and sensory expert evaluation was carried out in six sessions during different harvesting times in the black truffle season (December, January, and February). Both techniques were able to separate truffles depending on the harvesting time. Also, a list of volatile organic compounds related to the aromatic attributes was reported. This information will help to provide a more objective T. melanosporum truffle sensory evaluation.

Comprehensive Characterization of Tuber maculatum, New in Uruguay: Morphological, Molecular, and Aromatic Analyses.

Although only a few species of Tuber account for the major truffle sales volume, many species that are not considered delicacies are finding their way to the market, especially in regions where the traditionally appreciated ones do not occur. This is the case for whitish truffles. Specimens of whitish truffles were collected in pecan (Carya illinoinensis) orchards in Uruguay in October 2021. Morphological and molecular methods were used to characterize and assess their identity as Tuber maculatum Vittad. An SPME extraction of volatile compounds and GC-MS analyses were performed to characterize the aromatic profile of these specimens and evaluate their potential applications. Among the 60 VOCs detected, 3-octenone (mushroom odor), 3-octanol (moss, nut, mushroom odor), and 2H-pyran-2-one (no odor), followed by octen-1-ol-acetate (no odor) and 2-undecanone (orange, fresh, green odor) were the major compounds in T. maculatum fruiting bodies. The attributes of exotic edible mushrooms of commercial value in the region are highlighted. In particular, this work emphasizes the characteristics of truffles as a byproduct of pecan cultivation.

Extraction and trapping of truffle flavoring compounds into food matrices using supercritical CO2.

A supercritical fluid extraction methodology was used to extract flavoring and bioactive compounds from truffles. Some parameters such as CO2 flow rate (1-3 mg/mL), extraction time (15-90 min) and different trapping food matrices (grape seed oil, gelatin, agar agar and water) were optimized using response surface methodology to enhance extraction and trapping yields. The optimal conditions (2.27 mg/mL CO2 flow rate, 82.5 min when using 40 °C and 30 MPa, with 1 mL grape seed oil as trapping matrix) obtained with Tuber melanosporum were applied to three different truffle species: Terfezia claveryi, Tuber aestivum and Tuber indicum. A total of 32 metabolites were profiled in the extracts using ultra-high-performance supercritical fluid chromatography coupled to quadrupole time-of-flight mass spectrometry. Compounds such as brassicasterol ergosta-7,22-dienol, oleic and linoleic acid were found at similar amounts in all the extracts but other molecules (e.g. fungal sterols) showed a particular distribution depending on the specie studied and whether a trapping matrix was used at the SFE outlet.

Aromatic changes in home-made truffle products after heat treatments.

Truffles are highly valued by their aromatic properties and can aromatize food products. However, the truffle aroma could be reduced or lost with heat treatments (pasteurization and sterilization) necessary for products security and safety. In this study, sunflower oil and honey were aromatized with black truffle (lyophilized and fresh) using two different concentrations (5 and 10 %) for 24 h and then heat treatments (pasteurization and sterilization) were carried out. Truffle organic volatile compounds from products were investigated by SPME-GC-MS and sensory analysis by trained panel. More than 80 compounds were detected. Some of them were affected differently by heat process depending on the food matrix. Professional tasters scored higher key aromatic attributes such as sulphurous and olive oil in fresh truffle products, regardless the heat treatment applied.

Black truffle aroma transfer kinetics to food matrices.

Nowadays black truffles are so highly valued that truffled products are available in supermarkets whereas fresh truffle is mainly used in the restaurants. It is known that truffle aroma can change because heat treatments, but there is no scientific evidence about what molecules are transferred, in which concentration, and how much time is needed to aromatize products with truffle. In this study, four different fat-based food products (milk, sunflower oil, grapeseed oil and egg's yolk), were used to study black truffle (Tuber melanosporum) aroma transference for 14 days. Gas chromatography and olfactometry results showed different volatile organic compounds profile depending on the matrix used. After 24 h, some key truffle aromatic compounds were detected in all the food matrices. Among them, grape seed oil was the most aromatized product probably because of its odorless properties. According to our results, dimethyl disulphide, 3-methyl-1-butanol and 1-octen-3-one odorants showed the highest aromatization power.

Pressurized Liquid (PLE) Truffle Extracts Have Inhibitory Activity on Key Enzymes Related to Type 2 Diabetes (α-Glucosidase and α-Amylase).

An optimized PLE method was applied to several truffle species using three different solvent mixtures to obtain bioactive enriched fractions. The pressurized water extracts contained mainly (1 → 3),(1 → 6)-ß-D-glucans, chitins, and heteropolymers with galactose and mannose in their structures. The ethanol extracts included fatty acids and fungal sterols and others such as brassicasterol and stigmasterol, depending on the species. They also showed a different fatty acid lipid profile depending on the solvent utilized and species considered. Ethanol:water extracts showed interesting lipids and many phenolic compounds; however, no synergic extraction of compounds was noticed. Some of the truffle extracts were able to inhibit enzymes related to type 2 diabetes; pressurized water extracts mainly inhibited the α-amylase enzyme, while ethanolic extracts were more able to inhibit α-glucosidase. Tuber brumale var. moschatum and T. aestivum var. uncinatum extracts showed an IC50 of 29.22 mg/mL towards α-amylase and 7.93 mg/mL towards α-glucosidase. Thus, use of the PLE method allows o bioactive enriched fractions to be obtained from truffles with antidiabetic properties.

Aromatic profile of black truffle grown in Argentina: Characterization of commercial categories and alterations associated to maturation, harvesting date and orchard management practices.

Black truffle (Tuber melanosporum) is one of the most appreciated fungi in the world mainly due to its aromatic properties. In the emerging markets such as Argentina, the aroma of locally produced truffles has not been described yet. The volatile organic compounds (VOCs) from 102 black truffles from Argentina were analyzed using solid phase microextraction gas chromatography coupled with mass spectrometer detector (SPME-GC-MS). Several factors such as commercial category, maturity stage, host tree, geographical origin, and aromatic defects detected during classification were also registered and considered. As a result, 79 VOCs were detected, among which 2-methyl-propanal, 2-butanone, 2-methyl-1-propanol, butanal-3-methyl, 3-methyl-1-butanol, 2-methyl-1-butanol were present in high percentage in fresh mature truffles, whereas immature truffles were associated with 3,5-dimethoxytoluene, 2-phenyl-2-butenal, 2,3-dimethoxytoluene. The Argentine black truffles showed significant similarities in their aromatic profile when compared with their Australian and European counterparts, but with some distinctive notes.

Odor Profile of Four Cultivated and Freeze-Dried Edible Mushrooms by Using Sensory Panel, Electronic Nose and GC-MS.

Cultivated mushrooms are well-known nutrient inputs for an equilibrated diet. Some species are broadly appreciated due to their medicinal properties. Lately, a number of novel foods and nutraceuticals based on dehydrated and freeze-dried powder obtained from cultivated mushrooms has been reaching the market. The food industry requires fast and reliable tools to prevent fraud. In this, work we have cultivated Agaricus bisporus sp. bisporus (AB) (white button mushroom), Agaricus bisporus sp. brunnescens (ABP) (portobello), Lentinula edodes (LE) (shiitake) and Grifola frondosa (GF) (maitake) using tailor-made substrates for the different species and standardized cropping conditions, which were individually freeze-dried to obtain the samples under evaluation. The aim of this article was to validate the use of two different methodologies, namely, electronic nose and sensory panel, to discriminate the olfactory profile of nutritional products based on freeze-dried mushrooms from the different cultivated species. Additionally, GC-MS was used to detect and quantify the most abundant volatile organic compounds (VOCs) in the samples. The multivariate analysis performed proved the utility of electronic nose as an analytical tool, which was similar to the classical sensory panel but faster in distinguishing among the different species, with one limitation it being unable to differentiate between the same species. GC-MS analysis showed the chemical volatile formulation of the samples, also showing significant differences between different samples but high similarities between varieties of the same cultivated species. The techniques employed can be used to prevent fraud and have the potential to evaluate further medicinal mushroom species and build solid and trustful connections between these novel food products and potential consumers.

Application of Pressurized Liquid Extractions to Obtain Bioactive Compounds from Tuber aestivum and Terfezia claveryi.

A PLE (pressurized liquid extraction) method was adjusted following a full-factorial experimental design to obtain bioactive-enriched fractions from Tuber aestivum and Terfezia claveryi. Temperature, time and solvent (water, ethanol and ethanol-water 1:1) parameters were investigated. The response variables investigated were: obtained yield and the levels of total carbohydrate (compounds, ß-glucans, chitin, proteins, phenolic compounds and sterols). Principal component analysis indicated water solvent and high temperatures as more adequate parameters to extract polysaccharide-rich fractions (up to 68% of content), whereas ethanol was more suitable to extract fungal sterols (up to 12.5% of content). The fractions obtained at optimal conditions (16.7 MPa, 180 °C, 30 min) were able to protect Caco2 cells from free radical exposure, acting as antioxidants, and were able to reduce secretion of pro-inflammatory cytokines in vitro: IL-6 (50%), and TNFα (80% only T. claveryi ethanol extract), as well as reduce high inhibitory activity (T. aestivum IC50: 9.44 mG/mL).

Chemical composition and evaluation of antioxidant, antimicrobial and antiproliferative activities of Tuber and Terfezia truffles.

Ten truffle species of Tuber and Terfezia genera were chemical characterized, assessing their proximate composition, individual nutrient compounds and some bioactive molecules. The bioactive properties of these species were also evaluated, namely their antioxidant, antimicrobial and cytotoxic potential. Carbohydrates were the main macronutrients present in truffles, followed by proteins. Furthermore, the levels of polyunsaturated fatty acids (PUFA), subsequently presented as a percentage, were higher in truffles (38.2-79.3%) except in Tuber magnatum and Terfezia arenaria, which have a more saturated fatty acids (SFA) profile (70.7% and 53.7%, respectively). Comparing the species, T. magnatum revealed the highest levels of total phenolic compounds (TPC) (290 mg GAE/100 g truffle), as also the best results in the four methods used to evaluate the antioxidant activity. On the other hand, only five extracts obtained from some studied truffle species (Terfezia magnusii, Tuber aestivum, Tuber gennadii, and Tuber melanosporum) showed a slight inhibition of microbial growth, tested against different bacteria. Terfezia and T. gennadii extracts, showed potential to inhibit the cellular growth of NCI-H460, HeLa, HepG2, and MCF-7 cell lines (GI50 concentrations range: 19-78, 33-301, 83-321 and 102-321 µg/mL, respectively), indicating anti-proliferative activity. Nevertheless, T. arenaria revealed some potential hepatotoxicity, inhibiting the growth of PLP2 cells (GI50 concentration of 220 µg/mL), a primary cell culture obtained from porcine liver.

Edaphic and temporal patterns of Tuber melanosporum fruitbody traits and effect of localised peat-based amendment.

In Tuber melanosporum cultivation, fruitbody traits are gaining relevance due to their increasing prominence on prices. We investigated the edaphic and temporal patterns of fruitbody traits and characterised the effect of truffle nests (localised peat-based amendment supplemented with T. melanosporum spores) on traits. We monitored fruitbody traits throughout two fruiting seasons in three blocks along a soil gradient. Each trait followed specific edaphic and temporal patterns. The number of fruitbodies per dig and spore maturity followed characteristic within-season trends, whereas fruitbody weight and infestation by truffle beetles were subject to complex interactions among edaphic and temporal variables, suggesting a relevant influence of annual environmental conditions. The application of truffle nests increased fruitbody depth, improved its shape and decreased infestation by truffle beetles. Nests increased the number of fruitbodies per dig, but only in two of the soils, suggesting a relevant role of the bulk soil/substrate interface in fruiting initiation. These results outline a complex scenario, with each trait being differently affected by environmental factors. In this scenario, nests proved to effectively modify several traits, although not always in the desired direction.

Effects of gamma irradiation on the shelf-life and bioactive compounds of Tuber aestivum truffles packaged in passive modified atmosphere.

The effects of gamma irradiation (0.5, 1.0, 1.5 and 2.5 kGy doses) on Tuber aestivum packaged under modified atmosphere was evaluated. The respiration rate, microbial populations, sensory characteristics and content of bioactive compounds (total carbohydrates, chitins, ß-glucans, proteins, total phenols and sterols) were monitored from immediately after treatment up to day 42 of storage at 4 °C. All the irradiation treatments tested reduced the microbial groups studied by more than 3 log cfu/g. Increasing irradiation doses slowed down the subsequent microbial development throughout the conservation period for all the groups studied. The irradiation treatments did not negatively affect truffle sensory characteristics. Only a slight visible superficial yeast growth was detected at the end of the shelf-life in all doses applied. Total carbohydrate content, chitins, ß-glucans and proteins levels were not affected after irradiation. However, sterols, particularly stigmasterol, slightly decreased after irradiation, while levels of phenolic compounds doubled during storage. Gamma irradiation (2.5 kGy) could be used to extend the shelf-life of summer truffles packaged under modified atmosphere, since no remarkable reduction of bioactive compounds were noticed after 42 days of storage, and their sensory and microbial parameters were of higher quality than those of non-irradiated controls.

Screening of bioactive compounds in truffles and evaluation of pressurized liquid extractions (PLE) to obtain fractions with biological activities.

Truffles, besides the appreciated aromatic compounds, contain other molecules with interesting bioactive properties. A screening of fungal sterols and ß-glucans within different truffle species and locations was carried out. These compounds were extracted with pressurized liquids (PLE) generating enriched fractions. Extraction efficiency was studied with a full-factorial experimental design (Response surface methodology, RSM), using water and ethanol as extraction solvents. Polysaccharides from truffle powder (TP) and the optimal PLE extract (EP) obtained were precipitated and analysed by NMR and GC-MS. THP-1 cell cultures were utilized to test immunomodulatory properties. With the optimal PLE conditions (16.7 MPa, 180 °C, 30 min) 64 and 22.5% yields were obtained respect, with water and ethanol, generating fractions containing respect, 9.1% ß-glucan and 4.5% ergosterol. NMR analyses detected (1 â†’ 3)-ß-glucan structures in truffle. The EP induced a reduction of 40% IL-1ß and 60% IL-6 pro-inflammatory cytokines secretion suggesting potential immunomodulatory activity.

In vitro and in vivo testing of the hypocholesterolemic activity of ergosterol- and ß-glucan-enriched extracts obtained from shiitake mushrooms (Lentinula edodes).

Herein, a supercritical extraction plant (with a 6 L extraction cell) was successfully used to obtain ergosterol-enriched extracts from Lentinula edodes under the following conditions: a temperature of 40 °C, pressure of 225 bar, reaction time of 1-5 h, and the flow rate of 20 L h-1 for recirculated CO2. Moreover, ergosterol (ERG) and the SFE extract (SFE) with highest ergosterol concentration were microemulsified and submitted to in vitro digestion to study their ability to displace cholesterol from dietary mixed micelles (DMMs). ERG was also mixed with a ß-glucan-enriched (33.5%) extract (BGE) obtained from L. edodes to investigate the synergies between them; the results indicated that all these extracts (including BGE without ERG) could reduce the cholesterol levels in the DMMs. However, when ERG and SFE were simultaneously administered to mice with a hypercholesterolemic diet, no significant differences in the serum cholesterol levels were detected as compared to the case of the control. However, when only BGE was administrated to another mice model previously induced with hypercholesterolemia, significant reduction in the cholesterol levels was noticed.