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Article

Volatile Flavor of Tricholoma matsutake from the Different Regions of China by Using GC×GC-TOF MS

by
Yunli Feng
1,†,
Shaoxiong Liu
1,†,
Yuan Fang
2,
Jianying Li
1,
Ming Ma
1,
Zhenfu Yang
1,
Lue Shang
1,
Xiang Guo
1,
Rong Hua
1,2,* and
Dafeng Sun
1,2,*
1
Kunming Edible Fungi Institute of All China Federation of Supply and Marketing Cooperatives, Kunming 650221, China
2
Yunnan Academy of Edible Fungi Industry Development, Kunming 650221, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Foods 2025, 14(10), 1824; https://doi.org/10.3390/foods14101824
Submission received: 18 April 2025 / Revised: 15 May 2025 / Accepted: 17 May 2025 / Published: 21 May 2025
(This article belongs to the Section Food Analytical Methods)
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Abstract

:
Two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOF MS) was employed to analyze the volatile flavor compounds (VOCs) of Tricholoma matsutake samples from six different geographical regions: CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin) and XZ (Linzhi). The result indicate that a total of 2730 kinds of VOCs were identified from the fruiting bodies of six T. matsutake samples. The primary types of volatile organic compounds identified were 349 alcohols, 92 aldehydes, 146 carboxylic_acids, 311 esters, 742 organoheterocyclic compounds, 630 hydrocarbons, 381 ketones, 51 organic acids, and 28 derivatives and organosulfur compounds. Furthermore, PCA and PLS-DA analysis from the GC×GC-ToF-MS showed that samples from different regions could be distinguished by their VOCs. Network analysis revealed that 33 aroma compounds were identified as markers for distinguishing the samples from the six regions. The sensory attributes sweet, fruity, green, waxy, and floral were found to be more significant to the flavor profile of T. matsutake. 1-Nonanol, 2-Nonanone, Nonanoic acid, ethyl ester, 1-Undecanol, 2-Undecanone, Octanoic acid, ethyl ester, 2H-Pyran, and tetrahy-dro-4-methyl-2-(2-methyl-1-propenyl)- primarily contribute to the differences in the aroma characteristics among six T. matsutake samples. The results also provide a theoretical and practical foundation for the flavor compounds of these precious edible fungi in different regions.

1. Introduction

Tricholoma matsutake (S. Ito & S. Imai) Singer (T. matsutake) is a well-known wild edible mushroom, distributed in Asia, Europe, and North America [1]. In East Asia, specifically China, Japan and Korea, T. matsutake is renowned for its distinctive flavor, making it a sought-after ingredient in these culinary traditions. This mushroom has been found to contain high concentrations of proteins and amino acids. In addition, minerals, unsaturated fatty acids, vitamins, dietary fiber and a variety of other nutrients have been identified within it [2,3]. Furthermore, it also contains polysaccharides, polysaccharide–protein complex fractions and other bioactive components, which have demonstrated significant medicinal and nutritional value [4,5]. Despite its popularity, it is yet to be cultivated artificially, and all of the T. matsutake fresh fruiting bodies come from the wild.
The primary distribution regions of T. matsutake in China are Yunnan, Sichuan, Tibet and Jilin. Geographical isolation has been shown to be associated with genetic variations, differential nutritional value and divergent flavor characteristics [5,6]. Volatile flavor compounds have been used as chemical indicators for the identification and classification of specific species [6,7]. In the previous investigations, a total 66 VOCs were identified from Sichuan (Dechang, Daofu, Maerkang) by GC-MS [6]. Besides Sichuan, there are many other regions in China that have T. matsutake, such as Yunnan, Xizang, and Jilin. The evaluation of the volatile substances of T. matsutake from different regions is beneficial in understanding the influence of environmental conditions on T. matsutake and providing a scientific basis for the industrial production of T. matsutake [6].
Two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOF MS) is a novel technology that combines GC with high resolution and sensitivity, enabling the detection of a greater number of volatile organic compounds. Due to its strong peak separation ability, it has been widely applied in the research of various foods, especially in separating numerous subtle volatiles in complex sample matrices [8], such as coffee [9], Baijiu [10], cream [11], and Chinese dry-cured hams [12]. With a remarkable spectral acquisition rate reaching 500 spectra per second, this system demonstrates exceptional throughput for resolving thousands of chemical constituents in time-critical analyses. Previous studies on palm and palmist oils have shown that GC×GC-ToF-MS detects more VOCs, particularly fatty acid methyl esters, than GC-MS [13].
So far, there has been no research on the volatile flavor compounds of T. matsutake fruiting bodies in different regions of China. The composition and content of volatile compounds are crucial in determining the unique aromas of food, and serve as important indicators for evaluating quality. In this study, our research group conducted an analysis of the volatile compounds by using GC×GC-TOF MS on T. matsutake fruiting bodies samples from six different geographical regions: CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin) and XZ (Linzhi). The purpose is to find the volatile substances that can distinguish T. matsutake from these places, and to provide the basis for the industrialization of T. matsutake.

2. Materials and Methods

2.1. Fungal Materials and Sample Production

The fruiting bodies of T. matsutake samples (Figure 1) were collected from six different geographical regions—Chuxiong (Yunnan province, China), Dali (Yunnan province, China), Diqing (Yunnan province, China), Yanji (Jilin province, China), Xiaojin (Sichuan province, China) and Linzhi (Xizang province, China)—in August of 2024, and were named CX, DL, DQ, JL, SC, and XZ, respectively. We collected three fruiting bodies from each region. The detailed information about these geographical regions is shown in Table 1. T. matsutake is selected based on uniform size, the absence of mechanical damage, and freedom from insect infestation. After removing the surface contaminants, T. matsutake samples were longitudinally sectioned into uniform slices of approximately 5 mm thickness and then stored in polyethylene cryotubes at a temperature of −80 °C for subsequent analyses. The experiments were performed over three replicates for each sample.

2.2. GC×GC-TOF MS Analysis

Analyses were performed using the LECO Pegasus® 4D instrument (LECO, St. Joseph, MI, USA), which consists of an Agilent 8890A GC (Agilent Technologies, Palo Alto, CA, USA) equipped with a split/splitless injector, a two-stage cryogenic modulator (LECO), and a time-of-flight mass spectrometry (TOFMS) detector (LECO). We weighed 0.5 g of each sample of T. matsutake and placed them into a 20 mL headspace vial. The sample was then extracted using solid-phase micro-extraction (SPME). The SPME was incubated at 80 °C for 10 min, extracted for 25 min, and desorbed at the GC injection port for 5 min. Subsequently, GC×GC-TOF MS analysis was performed. The first-dimensional chromatographic column was DB-Heavy Wax (30 m × 250 μm × 0.5 μm), and the second-dimensional chromatographic column was Rxi-5Sil MS (2 m × 150 μm × 0.15 μm) (Restek, Bellefonte, PA, USA). The carrier gas was high-purity helium (>99.999%), and the flow rate was set at 1.0 mL/min. The oven temperature program was as follows: initially maintain the oven temperature at 50 °C for 2 min, then increase to 230 °C at a rate of 5 °C/min and hold for 5 min. The secondary oven temperature was set 5 °C higher than that of the primary oven. The modulator temperature was always maintained at 15 °C above that of the second column. The modulator was operated with a 6.0 s modulation period. The GC injector temperature was set 250 °C. The final results of the volatile compounds were expressed as normalized intensity, and are available in Table S1.

2.3. Statistical Analysis

The flavor compounds of the samples were identified using the NIST2020 database and Chroma TOF software (version 4.44). Following the execution of comprehensive data analysis employing Chroma TOF software, the following information was obtained: the nomenclature of each compound, the retention times, the CAS numbers, the RI information from the database, the actual RI calculated for normal alkanes C7-C30, and the peak areas of the samples. The final analysis result was derived by integrating these interpretive pieces of information. To facilitate the comparison of data across different scales, the original data underwent total peak area normalization or internal standard method normalization processing [14]. This project used internal standards for standardization. Principal component analysis (PCA) and partial least squares–discriminant analysis (PLS-DA) were performed using SIMCA-P (v13.0) and the R language ropls package [15]. All flavor compounds in nontargeted flavor omics and sensory annotation were obtained from the following databases: Odor database (Odor Thresholds for Chemicals with Established Health Standards, 2nd Edition) and Flavordb database (https://cosylab.iiitd.edu.in/flavordb/) (accessed on 18 December 2024).

3. Results

3.1. Statistical Identification of VOCs in Six T. matsutake Samples from Different Regions Using GC×GC-ToF MS

The VOCs of six T. matsutake samples from different regions are shown in Figure 2, Table S1. The identification of VOCs has revealed that over 900 VOCs were observed in each sample. The VOCs count in JL (Yanji) was the highest, at 1234, and that in DL (Dali) was the lowest at 942. Figure 2B indicates that there were significant differences among the six T. matsutake samples. These results indicate that there are differences in VOCs among six T. matsutake regions. This difference may be closely related to its unique geographical environment. Yanji is located in the central region of the Changbai Mountain, and due to its high-latitude location, the average annual temperature in the region is low, ranging between 3 and 5 °C. These low-temperature climatic conditions result in a comparatively longer growth cycle for T. matsutake in this region, which may have promoted the accumulation of secondary metabolites, thereby affecting types and relative contents of volatile compounds in T. matsutake [1,6].

3.2. Analysis of VOCs Obtained by GC×GC-ToF MS

The detailed information on the VOCs identified by GC×GC-ToF MS is presented in Figure 3, Table S1. The six T. matsutake samples contained 2730 VOCs, including nine classes (Figure 3), i.e., alcohols (349), aldehydes (92), carboxylic_acids (146), esters (311), organoheterocyclic compounds (742), hydrocarbons (630), ketones (381), organic acids and derivatives (51) and organosulfur compounds (28).
As far as alcohols are concerned, JL (Yanji) and XZ (Linzhi) had the highest alcohol contents, both at 151, as indicated in Figure 3, Table S1. These were followed by SC (Xiaojin) (145), CX (Chuxiong) (144), DQ (Diqing) (137), and DL (Dali) (131). Alcohols can be divided into saturated alcohol and unsaturated alcohol; the saturated alcohol threshold is high, while the unsaturated alcohol threshold is relatively low, and it has unique odor, so it contributes more to the flavor [16,17,18]. The key alcohols of fresh T. matsutake were 3-Octanol, (E)-2-Octen-1-ol, 1-Octen-3-ol, 1-Octanol, 1-Hexanol, 1-Butanol and 3-methyl- [19,20]. 1-Octen-3-ol was present at relatively high levels in all T. matsutake samples (Table S1), with the highest relative content in the DL (Dali) sample at 117.42 and the lowest in the XZ (Linzhi) sample at 30.09, compared to other T. matsutake samples, which ranged from 50.41 to 62.19. 1-Octen-3-ol is known as mushroom alcohol and it has a strong, aromatic odor; decreases in concentrations will influence the odor characteristics of T. matsutake [6]. 1-Octen-3-ol is the product of linoleic acid in edible fungi, produced through autooxidation or enzymatic oxidation and cleavage [1,21,22]. 3-Octanol has a mushroom and moss aroma, and 1-Octanol has a chemical and sweet flavor [3].
The group of aldehydes consisted of a total of 92 VOCs (Table S1), which are found in SC (Xiaojin) (64), CX (Chuxiong) (60), JL (Yanji) (59), XZ (Linzhi) (58), DQ (Diqing) (55) and DL (Dali) (46). Aldehydes are known for their floral and fruity odor [23]. Acetaldehyde, Heptanal, Hexanal, Nonanal, Octanal, Pentanal, Butanal, 2-methyl-, Butanal, 3-methyl-, propanal and 2-methyl- were the main VOCs in all T. matsutake samples. Aldehydes are volatile flavor substances with a high content and relatively low odor threshold in edible fungi, which often significantly influence the characteristic flavor of the overall volatile profile [24,25]. They are the primary degradation products of lipid oxidation, although some can also be produced through Maillard-induced amino acids degradation [3,26]. Butanal, 3-methyl-, a branched Strecker aldehyde, was abundant in all T. matsutake samples, and it has a chocolate aroma. Hexanal has a fresh grass aroma, propanal, 2-methyl- has grassy and spicy aroma, Butanal, 2-methyl- has a coffee and nut aroma, Pentanal has a fermented bread and nut aroma, and Heptanal has a wine flavor [23,27].
Carboxylic acids originate from the hydrolysis of fats into short-chain volatile fatty acids, or the degradation of amino acids [28]. Table S1 shows that the major carboxylic acids were acetamide, N-2-propenyl-, propanoic acid, 2-methyl-, propanoic acid, 2,2-dimethyl-, anhydride with diethylborinic acid, Acetic acid, [(aminocarbonyl)amino]oxo-, and 1-Methylcyclopropanecarboxylic acid. They were the most abundant in DQ (Diqing) (66), and least present in DL (Dali) (38).
Esters usually add fruity and floral flavors to mushrooms, and they are generally produced by non-enzymatic catalyzed reactions of alcohols and organic acids or enzyme-catalyzed reactions involving microorganisms [29]. In six T. matsutake samples, 2-Propenoic acid, butyl ester, 2-Propenoic acid, ethyl ester, acetic acid, octyl ester, Benzenepropanoic acid, ethyl ester, Benzenepropanoic acid, methyl ester, Butanoic acid, 1-ethenylhexyl ester, Butanoic acid, ethyl ester, Decanoic acid, ethyl ester, Dodecanoic acid, ethyl ester, Dodecanoic acid, methyl ester, Ethyl Acetate, Ethyl formate, Formic acid, octyl ester, Heptanoic acid, ethyl ester, Heptanoic acid, methyl ester, Hexadecanoic acid, ethyl ester, Nonanoic acid, ethyl ester, Pentadecanoic acid, ethyl ester, Tetradecanoic acid and ethyl ester were the common representatives. The most abundant ester in six T. matsutake samples (Table S1) was Hexadecanoic acid, ethyl ester, which contributes a waxy, fruity and creamy aroma. It was particularly highly prevalent in CX (Chuxiong), at 55.34, and least in XZ (Linzhi), at 1.78.
Organoheterocyclic compounds were the predominant VOCs in six T. matsutake samples, including furans, furanones, pyrazine, pyranones and so on. They were the most abundant in JL (Yanji) (332) and the lowest in DL (Dali) (244). Pyrazine is a specific product of the Maillard reaction, and has a unique fragrance [30]; it is present as Pyrazine, Pyrazine, 2,5-dimethyl-, Pyrazine, ethyl-, and Pyrazine, methyl-. The most abundant furans in T. matsutake samples was 2-n-Butyl furan, which has a light fruit aroma, with sweet wine flavor.
Alkanes in hydrocarbons are primarily derived from the homolytic cleavage of the alkoxy radical of fatty acids, with usually a high threshold and no obvious flavor characteristics [31]. They do not significantly contribute to the flavor of T. matsutake, but they can complement each other with other aromatic substances to give T. matsutake # different flavors. Studies indicate that alkanes and alcohols can be converted into each other and change the sensory properties of foods, after which they can play a role in harmonizing and complementing the flavor of T. matsutake [19]. In our study, the amount of SC (Xiaojin) (281) was the highest, followed by JL (Yanji) (277), and DQ (Diqing) (229) was the lowest. Among them, the hydrocarbons with higher contents were Tetradecane, Pentadecane, Hexadecane, Heptadecane, Octadecane, Cyclohexane-d12, 1,3-Octadiene, and 2-Hexene, 3,5,5-trimethyl-.
Ketones are primarily produced by the oxidation of unsaturated lipids and Maillard reactions, and their thresholds are higher than those of aldehydes, among which short-chain ketones contribute fatty and burnt aroma notes, whereas their long-chain ketones contribute floral notes [27]. It is evident that the majority of ketones are characterized by relatively low odor thresholds, consequently exerting a more pronounced impact on the overall flavor profile [32]. The major ketones in the six T. matsutake samples were Acetone, 1-Octen-3-one, 2,3-Hexanedione, 2-Decanone, 2-Heptanone, 3-Heptanone, 2-Octanone, 3-Octanone, 2-Pentadecanone and 2-Undecanone. 1-Octen-3-one was the primary mushroom flavor substance found in fresh T. matsutake. It was particularly high in CX (Chuxiong) at 76.14, and lowest in XZ (Linzhi) at 0.04. 3-Octanone also contributes to the mushroom and butter taste of T. matsutake [19]. 2-Undecanone is characterized as a typical compound for fruity and green flavors, and is commonly associated with the flavor profiles of certain foodstuffs. Acetone has apple and pear flavors [23].
The formation of acids may occur as secondary reaction products of the thermal decomposition and thermal degradation of unsaturated fatty acids during drying [33]. In the six T. matsutake samples, there were only 51 organic acids and derivatives; the largest amount was in XZ (Linzhi) (26), followed by JL (Yanji) (23), DQ (Diqing) (21) and DQ (Diqing) (18), and CX (Chuxiong) (17) contained the least. The major organic acids and derivatives were Butanoic acid, 2-oxo-, Ethyl acetoacetate and Propanoic acid, 2-oxo-.
Organosulfur compounds are usually present in the volatile flavors of edible mushrooms. They often typically emit a strong, pungent odor [34]. It is worth noting that Dimethyl disulfide and Dimethyl trisulfide are the predominant flavor compounds of Lentinula edodes [35]. The presence of distinct odor compounds in various types of edible mushrooms gives rise to significantly different aroma characteristics. Table S1 shows that 28 organosulfur compounds were detected across the six T. matsutake samples. CX (Chuxiong) was found to have the highest abundance of organosulfur compounds (13), followed by SC (Xiaojin) (12), while the lowest was DL (Dali) (7). All T. matsutake samples contained Disulfide, dimethyl, which was confirmed to contribute to the characteristic flavor of T. matsutake.

3.3. PCA and PLS-DA Results of Six T. matsutake Samples in Different Regions

The PCA results of volatiles detected in the present study are shown in Figure 4A. In total, 888 volatile compounds were selected for analysis in the experiment. The PCA score plot distinctly shows the significant regional differentiation among the six T. matsutake samples based on their volatile flavor compounds. PCA is a widely utilized multivariate statistical analysis tool in the field of sample variance analysis. The primary advantage of PCA is its ability to simplify data and elucidate the interrelationships among different samples [36]. In our study, the contribution rates of PC1 and PC2 were 15.3% and 9.3%. The total contribution rates of PC1 and PC2 reached 24.6%. The distribution diagram indicates that six T. matsutake samples could be distinguished by PC1 and PC2, revealing distinct flavor relationships among the selected samples. Figure 4A indicates that two T. matsutake samples (DQ (Diqing), XZ (Linzhi)) were located close to each other. This may be because the geographical locations of the two places are relatively close, and the similarities of their environmental conditions, including climate and soil, lead to the formation of similar flavors. The geographical location and altitude differences of different producing areas resulted in a variety of climates and ecological environments, as a result of which T. matsutake presented different flavor characteristics.
Following the identification of distinct clustering patterns among the six T. matsutake samples through PCA, a systematic investigation was subsequently performed using PLS-DA to elucidate the key variables responsible for the observed inter-sample differentiations. As shown in Figure 4B, samples from different regions can be effectively distinguished using PLS-DA.

3.4. Different Flavor Substances of Six T. matsutake Samples in Different Regions

To analyze the data obtained from GC×GC-TOF MS, a nonparametric test (specifically, one-way ANOVA) was utilized to analyze the identified 888 flavor compounds. The findings indicate that 168 flavor compounds exhibited significant variations among the six T. matsutake samples, suggesting their potential as crucial markers for differentiating regional samples (Figure 5). The relative contents are shown by the different colors in the figure. The more red the color, the higher the relative content, and the more blue, the lower the relative content. These markers included 26 alcohols, 15 aldehydes, 10 carboxylic_acids, 24 esters, 39 organoheterocyclic compounds, 24 hydrocarbons, 25 ketones, 3 organic acids and derivatives and 2 organosulfur compounds. VOCs of the six T. matsutake samples from different regions exhibited both common peaks and characteristic peaks, indicating that the compositions and contents of T. matsutake flavor compounds from different regions were both similar and different. By systematically analyzing the heatmap, we can further investigate the influences of geographical variations on the flavor compounds of T. matsutake, thus providing a scientific basis for subsequent research (such as production area tracing and quality regulation).
The samples of DL (Dali), DQ (Diqing) and XZ (Linzhi) were clustered together, which may be related to their similar geographical environments. There were significant differences in flavor compounds between CX (Chuxiong), JL (Yanji) and SC (Xiaojin). Compared with the samples from DL (Dali), DQ (Diqing) and XZ (Linzhi), in the SC (Xiaojin) sample, 2-Undecano, 4-Penten-2-one, 3-methyl-, 3-Octanol, acetate, Bicyclo[2.2.2]oct-5-en-2-one, Ethanone, (3aR,4R,8R,8aS)-3a,4,7,8a-Tetramethyl-1,2,3,3a,4,5,8,8a-octahydro-4,8-methanoazulene, 1-(2-hydroxy-5-methoxyphenyl)-, 5-Cyclopropyl-2H-1,2,3,4-tetrazole, Cyclopentane, nonyl-, 2a,4a,6a,6b-Tetrahydrocyclopenta[cd]pentalene, Resorcinol, 2-acetyl-, 3,5-Diamino-1,2,4-triazole, Quinoline, 8-ethyl-, Cyclopropyl carbinol, 2(3H)-Furanone, dihydro-5,5-dimethyl-4-(3-oxobutyl)-, 1-(1,2-Dimethyl-cyclopent-2-enyl)-ethanone1-, 1-Propanol were higher. 2-Butenoic acid, 2-propenyl ester, 2-Tridecanone, 1-Undecanol, Ethanone, 1-(2,4-dihydroxyphenyl)-, 1,3-Propanediol, 2-(hydroxymethyl)-2-methyl-, Octane, 4,5-dimethyl-, 6-Tridecanone, 2,4-Octadienal, (E,E)-, Dibenzofuran, Nonanoic acid, ethyl ester, 3-Butyn-2-amine, 2-methyl-, 2-Pentadecyn-1-ol, a-Furil, 4-Nonanol, Propanoic acid, 2,2-dimethyl-, anhydride with diethylborinic acid, 9H-Xanthene, 1,7-Octadien-3-ol, 2,6-dimethyl-, 2,4,6-Octatrienal were higher in the CX (Chuxiong) sample. And 1H-Pyrazole-1-carboximidamide, 2-Undecen-4-ol, Crotonyl isothiocyanate, 2,4-Hexadienal, (E,E)-, Ethyl tridecanoate, 2H-Pyran, 3,4-dihydro-4-methyl-, Cinnoline, 4-ethyl-3-methyl-, 1-Undecene, Ethyl-2-benzofuran, 2,2′-Bi-1,3-dioxolane, Octadecanal, 1-Hexanol, 2-ethyl-, 4-Heptanol, 3-methyl-, 2-Hydroxy-4,6-dimethylbenzaldehyde, 3,5-di-tert-Butyl-4-hydroxybenzaldehyde, 1,3-Cyclobutanediol, 2,2,4,4-tetramethyl-, 1H-1,2,3-Triazole-4-carboxaldehyde and 3(2H)-Benzofuranone, 7-methyl- were higher in the JL (Yanji) sample. This result reveals the influence of geography and climate on the flavor of T. matsutake.

3.5. Analysis of Sensory Flavor Characteristics

The flavor profiles of products represent a complex composite of identifiable taste and odor features, as well as features that cannot be identified separately. In order to undertake a systematic analysis and comparison of flavor compounds, Flavordb [13] was employed as a sensory evaluation reference database. The possible flavor profiles of six T. matsutake samples are shown in Figure 6. In six T. matsutake samples, SC (Xiaojin) had the highest sweet, fruity, green, waxy and fatty flavors. The flavors of sweet, fruity, green, waxy, fatty and nutty were lowest in DL (Dali). The other flavors, such us fresh, floral, oily, herbal and nutty, were similar.
Network analysis is regarded as a powerful tool for the investigation of potential correlations between flavor compounds and sensory attributes [37]. As demonstrated in Figure 7, volatiles have been shown to have a significant impact on flavor profiles, as evidenced by studies involving nontargeted flavoromics and online flavor databases. The green circle signifies the sensory feature, while the red circle represents the flavor compound. The larger the green circle, the greater the variety of flavor compounds associated with sensory characteristics, indicating its increased importance. The larger the red circle, the greater the number of sensory characteristics linked to the flavor compound, signifying the greater significance of the flavor substance. As indicated by the network, the most significant flavor compounds that have been identified as potentially key contributors to the aroma attributes of T. matsutake are further emphasized. The results show that 33 flavor compounds had a significant correlation with ten sensory attributes, suggesting that these 33 flavor compounds are key contributors to the flavor variations among six T. matsutake samples from different regions. Among them, the sensory attributes sweet, fruity, green, waxy, fatty, fresh, floral, rose, wine, and soapy were significantly correlated with 13, 16, 14, 13, 7, 8, 14, 7, 5, and 5 kinds of flavor compounds, respectively (Figure 7). So the sensory attributes sweet, fruity, green, waxy, and floral were more important. In the network, there is an interplay between sensory attributes and flavor compoundsl for example, 1-Nonanol, 2-Nonanone, Nonanoic acid, ethyl ester, 1-Undecanol, 2-Undecanone, Octanoic acid, ethyl ester, 2H-Pyran and tetrahydro-4-methyl-2-(2-methyl-1-propenyl)- were associated with T. matsutake aroma notes. In summary, these flavor compounds are primary contributors to the differences in the aroma characteristics among six T. matsutake samples.

4. Conclusions

In this study, GC×GC-ToF MS was utilized to analyze the volatile flavor substances present in six T. matsutake samples. The total number of VOCs identified in this study was found to be 2730. The sample from JL (Yanji) exhibited the highest count of VOCs, at 1234, whereas the sample from DL (Dali) had the lowest count, at 942. The primary types of volatile organic compounds were identified for alcohols, aldehydes, carboxylic acids, esters, organoheterocyclic compounds, hydrocarbons, ketones, organic acids and derivatives, and organosulfur compounds. Furthermore, the PCA and PLS-DA analysis from the GC×GC-ToF MS indicates that samples from different regions could be distinguished based on their VOCs. The results show that the VOCs of DQ (Diqing) and XZ (Linzhi) are the closest, which may be because the geographical locations of the two regions are relatively close. Network analysis enabled the identification of 33 aroma compounds that function as markers to distinguish samples from six regions. The sensory attributes sweet, fruity, green, waxy, and floral were more important to the flavor of T. matsutake. These flavor compounds, 1-Nonanol, 2-Nonanone, Nonanoic acid, ethyl ester, 1-Undecanol, 2-Undecanone, Octanoic acid, ethyl ester, 2H-Pyran, and tetrahydro-4-methyl-2-(2-methyl-1-propenyl)-, are primarily responsible for the variations in the aroma characteristics of the six T. matsutake samples.
The information presented in this study offers significant new insights into the flavor profile of T. matsutake in China. The results can also provide a theoretical and practical foundation for the flavors of precious edible fungal in different regions. However, this study also has certain limitations. The number of samples is limited, involving only six regions’ samples, which may not fully reflect all the characteristics of the volatile flavor substances of T. matsutake in China. Future research can expand the sample range to include more regions and varieties of matsutake mushrooms so as to further enhance the representativeness and accuracy of the results.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/foods14101824/s1, Table S1: GC×GC-ToF MS detected VOCs of six T. matsutake samples from different regions.

Author Contributions

Conceptualization, Y.F. (Yunli Feng) and S.L.; methodology, Y.F. (Yunli Feng); software, Y.F. (Yuan Fang) and S.L.; validation, Y.F. (Yunli Feng), Y.F. (Yuan Fang) and M.M.; formal analysis, S.L. and Y.F. (Yuan Fang); investigation, X.G.; resources, J.L.; data curation, Y.F. (Yunli Feng); writing—original draft preparation, Y.F. (Yunli Feng) and S.L.; writing—review and editing, Y.F. (Yunli Feng) and S.L.; visualization, J.L.; supervision, Z.Y.; project administration, L.S.; funding acquisition, R.H. and D.S. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Yunnan Major Science and Technology Special Project Program (202402AE090001), the Yunnan Province Science and Technology Talents and platform plan (202205AD160042), the Yunnan Province integration project (202402AN360003), and the Yunnan Province Science and Technology Talents and platform plan (Academician expert workstation) (202305AF150187).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in the study are included in the article/Supplementary Material, further inquiries can be directed to the corresponding authors.

Acknowledgments

The authors would like to thank laboratory colleagues for their advice and support.

Conflicts of Interest

The authors declare no conflicts of interest.

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Foods 14 01824 g001
Figure 1. The fruiting bodies of T. matsutake.
Figure 1. The fruiting bodies of T. matsutake.
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Figure 2. Statistic of identified VOCs of six T. matsutake samples from different regions using GC×GC-ToF MS. (A) Statistics of identified VOCs of six T. matsutake samples from different regions. (B) Single-indicator multiple-comparison bar chart of volatiles of six T. matsutake samples. a, p < 0.05; b, p < 0.01. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 2. Statistic of identified VOCs of six T. matsutake samples from different regions using GC×GC-ToF MS. (A) Statistics of identified VOCs of six T. matsutake samples from different regions. (B) Single-indicator multiple-comparison bar chart of volatiles of six T. matsutake samples. a, p < 0.05; b, p < 0.01. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Figure 3. The relative contents of nine classes of VOCs in the six T. matsutake samples from different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 3. The relative contents of nine classes of VOCs in the six T. matsutake samples from different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Figure 4. Volatile organic compounds detected in six T. matsutake samples from different regions. (A) PCA of volatiles of six T. matsutake samples; (B) PLS-DA of volatiles of six T. matsutake samples. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 4. Volatile organic compounds detected in six T. matsutake samples from different regions. (A) PCA of volatiles of six T. matsutake samples; (B) PLS-DA of volatiles of six T. matsutake samples. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Figure 5. Cluster heatmap of volatile flavor compounds of six T. matsutake samples in different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 5. Cluster heatmap of volatile flavor compounds of six T. matsutake samples in different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Figure 6. Radar map illustrating the sensory flavor characteristics of six T. matsutake samples from different regions. The outermost circle represents the sensory flavor characteristics, and the broken line indicates the frequency level of the corresponding flavor substance (the detection frequency is classified from 1 to 5, with the highest frequency being level 5). CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 6. Radar map illustrating the sensory flavor characteristics of six T. matsutake samples from different regions. The outermost circle represents the sensory flavor characteristics, and the broken line indicates the frequency level of the corresponding flavor substance (the detection frequency is classified from 1 to 5, with the highest frequency being level 5). CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Figure 7. The correlation network between sensory attributes and aroma compounds (p < 0.05) of six T. matsutake samples from different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
Figure 7. The correlation network between sensory attributes and aroma compounds (p < 0.05) of six T. matsutake samples from different regions. CX (Chuxiong), DL (Dali), DQ (Diqing), JL (Yanji), SC (Xiaojin), XZ (Linzhi).
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Table 1. Tricholoma matsutake sampling sites in China.
Table 1. Tricholoma matsutake sampling sites in China.
NameCollection PlacesGeographic Position
CXNanhua, Chuxiong, Yunnan province, ChinaN 24°51′29″, E 100°48′32″
DLJianchuan, Dali, Yunnan province, ChinaN 26°35′9″, E 99°40′9″
DQShangri-La, Diqing, Yunnan province, ChinaN 27°54′14″, E 99°38′14″
JLYanji, Jilin province, ChinaN 42°51′1″, E 129°29′59″
SCXiaojin, Aba, Sichuan province, ChinaN 30°57′49″, E 102°17′58″
XZLinzhi, Xizang province, ChinaN 29°53′53″, E 93°26′42″
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Feng, Y.; Liu, S.; Fang, Y.; Li, J.; Ma, M.; Yang, Z.; Shang, L.; Guo, X.; Hua, R.; Sun, D. Volatile Flavor of Tricholoma matsutake from the Different Regions of China by Using GC×GC-TOF MS. Foods 2025, 14, 1824. https://doi.org/10.3390/foods14101824

AMA Style

Feng Y, Liu S, Fang Y, Li J, Ma M, Yang Z, Shang L, Guo X, Hua R, Sun D. Volatile Flavor of Tricholoma matsutake from the Different Regions of China by Using GC×GC-TOF MS. Foods. 2025; 14(10):1824. https://doi.org/10.3390/foods14101824

Chicago/Turabian Style

Feng, Yunli, Shaoxiong Liu, Yuan Fang, Jianying Li, Ming Ma, Zhenfu Yang, Lue Shang, Xiang Guo, Rong Hua, and Dafeng Sun. 2025. "Volatile Flavor of Tricholoma matsutake from the Different Regions of China by Using GC×GC-TOF MS" Foods 14, no. 10: 1824. https://doi.org/10.3390/foods14101824

APA Style

Feng, Y., Liu, S., Fang, Y., Li, J., Ma, M., Yang, Z., Shang, L., Guo, X., Hua, R., & Sun, D. (2025). Volatile Flavor of Tricholoma matsutake from the Different Regions of China by Using GC×GC-TOF MS. Foods, 14(10), 1824. https://doi.org/10.3390/foods14101824

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