Gas Chromatography Time-of-flight Mass Spectrometry Research Articles

Serum metabolomics signature of maternally inherited diabetes and deafness by gas chromatography-time of flight mass spectrometry.

The aim of this study was to identify a metabolic signature of MIDD as compared to healthy controls and other types of diabetes. We performed a comprehensive serum metabolomic analysis using gas chromatography-time of flight mass spectrometry (GC-TOFMS) in participants diagnosed with MIDD (n = 14), latent autoimmune diabetes in adults (LADA) (n = 14), type 2 diabetes mellitus (n = 14), and healthy controls (n = 14). Each group was matched for gender and age. There were significant metabolic differences among MIDD and other diabetic and control groups. Compared with control, MIDD patients had high levels of carbohydrates (glucose, galactose, mannose, sorbose, and maltose), fatty acids (2-Hydroxybutyric acid, eicosapentaenoic acid, and octadecanoic acid), and other metabolites (alanine, threonic acid, cholesterol, lactic acid, and gluconic acid), but low level of threonine. Compared with LADA, MIDD patients had high levels of threonic acid and some amino acids (alanine, tryptophan, histidine, proline, glutamine, and creatine) but low levels of serine. Compared with type 2 diabetes mellitus, MIDD patients had high levels of citrulline, creatine, 3-Amino-2-piperidone, but low levels of ornithine, fatty acids (arachidonic acid and octadecanoic acid), and intermediates of the tricarboxylic acid cycle (malic acid and succinic acid). Our study identified a specific metabolic profile related to glycolysis and the tricarboxylic acid cycle in MIDD that differs from healthy controls and other types of diabetes. This unique metabolic signature provides new perspectives for understanding the pathophysiology and underlying mechanisms of MIDD.

Molecular composition of Beijing PM2.5 Brown carbon associated with fluorescence revealed by gas chromatography time-of-flight mass spectrometry and parallel factor analysis

Brown carbon (BrC), the important component of carbonaceous aerosol, attracted much attention for its complex chemical composition which affect its light absorption, fluorescent properties, and atmospheric reactions. In this study, we revealed for the first time the molecular composition of non-polar/low-polar organic matters contributing to fluorescence of Beijing PM2.5 from 2016 to 2018. Firstly, excitation-emission matrix (EEM) spectra of the 122 PM2.5 samples were analyzed and supported by gas chromatography-time of flight mass spectrometry (GC-ToF-MS) and partial least squares regression model, 400 BrC molecules associated with fluorescence were screened out among which 44 were the newly ones. Secondly, six chromophores (C1–C6) were identified by applying parallel factor analysis (PARAFAC) model, which represented categories of compounds consistent with the ToF results. Then, the specific molecule composition of each chromophore was achieved by combining GC-ToF-MS and EEM-PARAFAC. For example, indeno[1,2,3-cd]pyrene is a new fluorescent molecule which contributed high in C3 (high oxygenated humic-like chromophore) and C5 (fluorescein chromophore). The results can, on one side, enrich the database of fluorescent molecular composition, which turns out to be similar as that of light-absorption; on the other side and more importantly, provide chemical characteristics of BrC fluorescent chromophores on the level of molecule.

Sensory and chemical characterization of chestnuts processed in different methods using instrumental analyses and the Check-all-that-apply method

Chestnuts are known for their unique sensory characteristics and nutritional value. However, the changes in their sensory attributes after processing remain unclear. This study used instrumental analysis and sensory evaluation combined with multivariate statistical analysis to investigate the effect of packaging and thermal sterilization procedures on the sensory characteristics of chestnuts. The results showed that the significant variations (p < 0.05) between the different processing methods for chestnuts were revealed via the texture analysis, headspace solid-phase microextraction gas chromatography × gas chromatography-time of flight-mass spectrometry (HS-SPME-GC×GC-TOF-MS), and check-all-that-apply (CATA) sensory evaluation. The packaging had a more significant influence on the sensory quality of the chestnuts than thermal sterilization procedures. The HS-SPME-GC×GC-TOF-MS identified 116 volatile compounds, and the partial least squares-discriminant analysis (PLS-DA) identified the 28 volatile compounds responsible for the similarities and differences among different processing methods of chestnuts. Principal component analysis (PCA) combined with texture analysis and the fingerprint of HS-SPME-GC×GC-TOF-MS, as well as the correspondence analysis (CA) analysis of CATA, could discriminate samples from different packaging of chestnuts. This study provides chestnuts processing recommendations for satisfying consumer preferences and predicting possible related quality changes.

Investigating Distinct Skin Microbial Communities and Skin Metabolome Profiles in Atopic Dermatitis.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder influenced by genetic predisposition, environmental factors, immune dysregulation, and skin barrier dysfunction. The skin microbiome and metabolome play crucial roles in modulating the skin's immune environment and integrity. However, their specific contributions to AD remain unclear. We aimed to investigate the distinct skin microbial communities and skin metabolic compounds in AD patients compared to healthy controls (HCs). Seven patients with AD patients and seven HCs were enrolled, from whom skin samples were obtained for examination. The study involved 16S rRNA metagenomic sequencing and bioinformatics analysis as well as the use of gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) to detect metabolites associated with AD in the skin. We observed significant differences in microbial diversity between lesional and non-lesional skin of AD patients and HCs. Staphylococcus overgrowth was prominent in AD lesions, while Cutibacterium levels were decreased. Metabolomic analysis revealed elevated levels of several metabolites, including hypoxanthine and glycerol-3-phosphate in AD lesions, indicating perturbations in purine metabolism and energy production pathways. Moreover, we found a positive correlation between hypoxanthine and glycerol-3-phosphate and clinical severity of AD and Staphylococcus overgrowth. These findings suggest potential biomarkers for monitoring AD severity. Further research is needed to elucidate the causal relationships between microbial dysbiosis, metabolic alterations, and AD progression, paving the way for targeted therapeutic interventions.

Qualitative and Quantitative Metabolite Comparison of Korean Traditional Alcoholic Beverages: Takju, Yakju, and Traditional-Soju.

With increasing interest in Korean foods and beverages, Korean traditional alcoholic beverages need to be studied. To characterize Korean traditional alcoholic beverages, we analyzed the metabolites of Takju, Yakju, and Traditional-Soju using 48 commercial products. We performed non-targeted metabolite profiling using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) and identified 33 significantly discriminant metabolites, including nine organic acids, three amino acids, and seven fatty acids, in the three types of alcoholic beverage. Subsequently, we quantified the profiled metabolites in each product and compared their contents to identify alcoholic beverage type-specific metabolites. Thus, we figured out seven metabolites using receiver operating characteristic (ROC) curves. The results revealed that octadecanoic acid (limit of detection (LOD) to 168.72 mg/L), nonanoic acid (LOD to 112.54 mg/L), and octanoic acid (8.00 to 145.08 mg/L) in Takju; succinic acid (LOD to 1.90 mg/mL), heptanoic acid (LOD to 343.23 mg/L), and hexadecanoic acid (20.28 to 126.45 mg/L) in Yakju; and malonic acid (LOD to 19.13 mg/mL) in Traditional-Soju, with an area under the curve (AUC) > 0.7, are important metabolites that can distinguish the type of alcoholic beverage. Our results provide qualitative and quantitative metabolite information about Korean traditional alcoholic beverages that can be used by consumers and manufacturers.

Lactiplantibacillus plantarum ZFM55 improves texture and flavor of yogurt, increases beneficial metabolites, and the co-fermented yogurt promotes human gut microbiota health

Yogurt offers potential health benefits, particularly for the intestines. The choice of starter culture is key to the production of functional yogurt. In this study, Lactiplantibacillus plantarum ZFM55 was incorporated into a commercial starter and its effects on the quality of yogurt was evaluated. Changes in texture, flavor, composition, and metabolites were assessed, along with its impact on the gut microbiota. We found adding ZFM55 improved the water holding capacity, cohesiveness, viscosity, and hardness of the yogurt. Analysis using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) and liquid chromatography-mass spectrometry (LC-MS) demonstrated that ZFM55 contributed to a desirable flavor profile and increased the levels of non-volatile metabolites, such as amino acids and carbohydrates. Stool samples from volunteers who consumed fermented milk were collected and analyzed using 16 S rDNA sequencing and GC-MS. The results showed that fermenting yogurt with ZFM55 significantly altered the beta diversity of the gut microbiota. Notably, probiotics such as Bifidobacterium, Blautia, and Lactobacillus exhibited substantial increases, along with a notable rise in the production of short-chain fatty acids, particularly acetic acid and butyric acid. These findings highlight the potential of ZFM55 for developing high-quality functional yogurt products.

Early Detection of Fusarium Basal Rot Infection in Onions and Shallots Based on VOC Profiles Analysis.

Gas chromatography ion-mobility spectrometry (GC-IMS) technology is drawing increasing attention due to its high sensitivity, low drift, and capability for the identification of compounds. The noninvasive detection of plant pests and pathogens is an application area well suited to this technology. In this work, we employed GC-IMS technology for early detection of Fusarium basal rot in brown onion, red onion, and shallot bulbs and for tracking disease progression during storage. The volatile profiles of the infected and healthy control bulbs were characterized using GC-IMS and gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS). GC-IMS data combined with principal component analysis and supervised methods provided discrimination between infected and healthy control bulbs as early as 1 day after incubation with the pathogen, classification regarding the proportion of infected to healthy bulbs in a sample, and prediction of the infection's duration with an average R2 = 0.92. Furthermore, GC-TOF-MS revealed several compounds, mostly sulfides and disulfides, that could be uniquely related to Fusarium basal rot infection.

Changes in Uterine Metabolome Associated with Metritis Development and Cure in Lactating Holstein Cows.

The objective of this study was to identify alterations in the vaginal discharge (VD) metabolome and potential biomarkers to predict metritis development and a cure in dairy cows. This prospective cohort study was conducted on two dairies located in CA and TX. Vaginal discharge was evaluated and collected using the Metricheck® device. Cows were examined for metritis at 4, 7, and 9 days in milk (DIM). Cows with a fetid, watery, and reddish-brown uterine discharge were classified as having metritis and randomized to receive ceftiofur (n = 10) or remain untreated (n = 7). A cure was defined as the absence of a fetid, watery, reddish-brown uterine discharge at 14 d after enrollment. Vaginal discharge samples were collected from 86 cows within 6 h after parturition, at 4 and 7 DIM, at metritis diagnosis, and at 4 and 7 days after metritis diagnosis. Cows with metritis (MET; n = 17) were paired with counterparts without metritis (HTH) of a similar DIM and parity (n = 34). The uterine metabolome was evaluated using untargeted gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). Metabolomic data were analyzed using the MetaboAnalyst 5.0. Data were log-transformed and auto-scaled for normalization. Univariate analyses, including the fold-change, were performed to identify the metabolites linked to metritis development and its cure and principal component analysis and partial least squares discriminant analysis were performed to explain metabolite variance between animals developing or not developing metritis and being cured or not being cured of metritis. Comparing HTH with MET cows at calving, 12 metabolites were upregulated, and one was downregulated. At four and seven DIM, 51 and 74 metabolites, respectively, were altered between MET and HTH cows. After metritis development, three and five metabolites were upregulated in cows that were cured and in cows that received treatment and were cured, respectively. In all scenarios, the metabolites lignoceric, malic, and maleic acids, ornithine, and hypotaurine, which are associated with arginine/aminoacyl-tRNA biosynthesis and taurine/purine metabolism, were upregulated in HTH cows. Metritis was associated with changes in the uterine metabolome. Cows not being cured of metritis had changes in the uterus metabolome independent of receiving ceftiofur or remaining untreated. Metabolome analysis may be an important tool to understand the vaginal discharge changes during postpartum and the dynamics of metritis development and cures and help to identify biomarkers to predict metritis being cured.

Structure and Metabolic Characteristics of Intestinal Microbiota in Tibetan and Han Populations of Qinghai-Tibet Plateau and Associated Influencing Factors.

Residents of the Qinghai-Tibet Plateau might experience shifts in their gut microbiota composition as a result of the plateau environment. For example, high altitudes can increase the abundance of obligate anaerobic bacteria, decrease the number of aerobic bacteria and facultative anaerobic bacteria, increase probiotics, and decrease pathogenic bacteria. This study aimed to determine the structure and metabolic differences in intestinal microbial communities among the Tibetan and Han populations on the Qinghai-Xizang Plateau and shed light on the factors that influence the abundance of the microbial communities in the gut. The structural characteristics of intestinal microorganisms were detected from blood and fecal samples using 16S rRNA sequencing. Metabolic characteristics were detected using gas chromatography-time-of-flight mass spectrometry (GC-TOFMS). The influencing factors were analyzed using Spearman's correlation analysis. Bacteroides and Bifidobacterium were dominant in the intestinal tract of the Han population, while Bacteroides and Prevotella were dominant in that of the Tibetan population, with marked differences in Pseudomonas, Prevotella, and other genera. Ferulic acid and 4-methylcatechol were the main differential metabolites between the Tibetan and Han ethnic groups. This may be the reason for the different adaptability of Tibetan and Han nationalities to the plateau. Alanine aminotransferase and uric acid also have a high correlation with different bacteria and metabolites, which may play a role. These results reveal notable disparities in the compositions and metabolic characteristics of gut microbial communities in the Tibetan and Han people residing on the Qinghai-Tibet Plateau and may provide insights regarding the mechanism of plateau adaptability.

Formation analysis of primary and secondary metabolites during angkak fermentation based on GC-TOF-MS, GC-FID, and HPLC and metabolomics analysis

The significant primary and secondary metabolites occurred during angkak fermentation are composed of organic acids, amino acids, monacolin K, pigments, and citrinin (a mycotoxin), which their variance is able to inform how the Monascus-fermented produce characteristics. Here, gas chromatography time of flight mass spectrometry (GC-TOF-MS), gas chromatography-flame ionization detector (GC-FID), and high-performance liquid chromatography (HPLC) were utilized for analysis of the metabolites in angkak (Monascus purpureus) for 30 days. Principal component analysis (PCA) and Pearson’s correlation coefficients were to study the relationship between the metabolites and fermentation time. Moreover, the metabolomics analysis in angkak was performed by using MetaboAnalyst 5.0. The amount of primary metabolites such as the organic acids (succinic acid, glycolic acid, oxalic acid, lactic acid, lactobionic acid, and pyruvic acid), amino acids (valine, isoleucine, and alanine), fatty acids (stearic acid, oleic acid, linoleic acid, palmitic acid, and linolenic acid), and, monosaccharides (i.e. fructose and glucose) increased gradually until 15 days of the fermentation and then would decrease (the death phase of M. purpureus). For the secondary metabolites, the levels of monacolin K, Monascus pigments, and citrinin (a mycotoxin)after day 10 of the fermentation increased more than 2 times as compared with the previous fermentation time of day 5, exposing to begin the stationary phase of M. purpureus growth. Similarly, according to the heat map of primary metabolites’ production rates, there were their higher production rates of primary metabolites after 10 and 15 days of the fermentation; whereas, the higher production rates of secondary metabolites were found after 25 and 30 days of the fermentation. The PCA and Pearson’s correlation coefficients explained that the primary and secondary metabolites were discriminated completely into different groups. Based on the data of 19 metabolites throughout 30 days of the fermentation, there were estimated 23 metabolisms, but outstanding 4 metabolisms were glycolysis, pyruvate metabolism, α-linolenic acid metabolism, and linoleic acid metabolism.

Comprehensive profiling of the metabolome in corn silage inoculated with or without Lactiplantibacillus plantarum using different untargeted metabolomics analyses

ABSTRACTSilage fermentation is a complicated biochemical process involving interactions between microbes and metabolites. However, the overall metabolome feature of ensiled forage and its response to lactic acid bacteria inoculation is poorly understood. Hence, in this study metabolome profiles of whole-plant corn silage inoculated with or without Lactiplantibacillus plantarum were characterised via solid-phase microextraction/gas chromatography/mass spectrometry (SPME-GC-MS), gas chromatography/time-of-flight mass spectrometry (GC-TOF-MS), and Liquid chromatography/Q Exactive HFX mass spectrometry (LC-QE-MS/MS) analysis. There were 2087 identified metabolites including 1143 reliably identified metabolites in fresh and ensiled whole-plant corn. After ensiling, the increased metabolites in whole-plant corn were mainly composed of organic acids, volatile organic compounds (VOC), benzene and substituted derivatives, carboxylic acids and derivatives, fatty acyls, flavonoids, indoles and derivatives, organooxygen compounds (including amines and amides), phenols, pyridines and derivatives, and steroids and steroid derivatives, which includes neurotransmitters and metabolites with aromatic, antioxidant, anti-inflammatory, and antimicrobial activities. Phenylacetaldehyde was the most abundant aromatic metabolite after ensiling. L-isoleucine and oxoproline were the major free amino acids in silage. Ensiling markedly increased the relative abundances of 3-phenyllactic acid, chrysoeriol, 6-O-acetylaustroinulin, acetylcholine, γ-aminobutyric acid, pyridoxine, and alpha-linoleic acid. Inoculation with L. plantarum remarkably changed silage VOC composition, and essential amino acids, 3-phenyllactic acid, and cinnamaldehyde compared with untreated silage. The present study does not only provide a deeper insight into metabolites of the ensiled whole-plant corn but also reveals metabolites with specific biological functions that could be much helpful in screening novel lactic acid bacteria to well ensile forages. Inoculation with L. plantarum significantly affects the metabolome in ensiled whole-plant corn.

The characteristic of the key aroma-active components in white tea using GC-TOF-MS and GC-olfactometry combined with sensory-directed flavor analysis.

White tea has become more and more popular with consumers due to its health benefits and unique flavor. However, the key aroma-active compounds of white tea during the aging process are still unclear. Thus, the key aroma-active compounds of white tea during the aging process were investigated using gas chromatography-time-of-flight-mass spectrometry (GC-TOF-MS) and gas chromatography-olfactometry (GC-O) combined with sensory-directed flavor analysis. A total of 127 volatile compounds were identified from white tea samples with different aging years by GC-TOF-MS. Fifty-eight aroma-active compounds were then determined by GC-O, and 19 of them were further selected as the key aroma-active compounds based on modified frequency (MF) and odor activity value (OAV). Aroma recombination and omission testing confirmed that 1-octen-3-ol, linalool, phenethyl alcohol, geraniol, (E)-β-ionone, α-ionone, hexanal, phenylacetaldehyde, nonanal, (E, Z)-(2,6)-nonadienal, safranal, γ-nonalactone and 2-amylfuran were the common key aroma-active compounds to all samples. Cedrol, linalool oxide II and methyl salicylate were confirmed peculiar in new white tea, while β-damascenone and jasmone were peculiar in aged white tea. This work will offer support for further studies on the material basis of flavor formation of white tea. © 2023 Society of Chemical Industry.

High expression of VARS promotes the growth of multiple myeloma cells by causing imbalance in valine metabolism.

Multiple myeloma (MM) is a plasma cell malignancy occurring in middle and old age. MM is still an incurable disease due to its frequent recurrence and drug resistance. However, its pathogenesis is still unclear. Abnormal amino acid metabolism is one of the important characteristics of MM, and the important metabolic pathway of amino acids participates in protein synthesis as basic raw materials. Aminoacyl transfer ribonucleic acid synthetase (ARS) gene is a key regulatory gene in protein synthesis. This study aims to explore the molecular mechanism for ARS, a key factor of amino acid metabolism, in regulating amino acid metabolism in MM and affecting MM growth. The corresponding gene number was combined with the gene expression profile GSE5900 dataset and GSE2658 dataset in Gene Expression Omnibus (GEO) database to standardize the gene expression data of ARS. GSEA_4.2.0 software was used to analyze the difference of gene enrichment between healthy donors (HD) and MM patients in GEO database. GraphPad Prism 7 was used to draw heat maps and perform data analysis. Kaplan-Meier and Cox regression model were used to analyze the expression of ARS gene and the prognosis of MM patients, respectively. Bone marrow samples from 7 newly diagnosed MM patients were collected, CD138+ and CD138- cells were obtained by using CD138 antibody magnetic beads, and the expression of ARS in MM clinical samples was analyzed by real-time RT-PCR. Human B lymphocyte GM12878 cells and human MM cell lines ARP1, NCI-H929, OCI-MY5, U266, RPMI 8266, OPM-2, JJN-3, KMS11, MM1.s cells were selected as the study objects. The expression of ARS in MM cell lines was analyzed by real-time RT-PCR and Western blotting. Short hairpin RNA (shRNA) lentiviruses were used to construct gene knock-out plasmids (VARS-sh group). No-load plasmids (scramble group) and gene knock-out plasmids (VARS-sh group) were transfected into HEK 293T cells with for virus packaging, respectively. Stable expression cell lines were established by infecting ARP1 and OCI-MY5 cells, and the effects of knockout valyl-tRNA synthetase (VARS) gene on proliferation and apoptosis of MM cells were detected by cell counting and flow cytometry, respectively. GEO data were divided into a high expression group and a low expression group according to the expression of VARS. Bioinformatics analysis was performed to explore the downstream pathways affected by VARS. Gas chromatography time-of-flight mass spectrometry (GC-TOF/MS) and high performance liquid chromatography (HPLC) were used to detect the valine content in CD138+ cells and ARP1, OCI-MY5 cells and supernatant of knockdown VARS gene in bone marrow samples from patients, respectively. Gene enrichment analysis showed that tRNA processing related genes were significantly enriched in MM compared with HD (P<0.0001). Further screening of tRNA processing-pathway related subsets revealed that cytoplasmic aminoacyl tRNA synthetase family genes were significantly enriched in MM (P<0.0001). The results of gene expression heat map showed that the ARS family genes except alanyl-tRNA synthetase (AARS), arginyl-tRNA synthetase (RARS), seryl-tRNA synthetase (SARS) in GEO data were highly expressed in MM (all P<0.01). With the development of monoclonal gammopathy of undetermined significance (MGUS) to MM, the gene expression level was increased gradually. Kaplan-Meier univariate analysis of survival results showed that there were significant differences in the prognosis of MM patients in methionyl-tRNA synthetase (MARS), asparaginyl-tRNA synthetase (NARS) and VARS between the high expression group and the low expression group (all P<0.05). Cox regression model multivariate analysis showed that the high expression of VARS was associated with abnormal overall survival time of MM (HR=1.83, 95% CI 1.10 to 3.06, P=0.021). The high expression of NARS (HR=0.90, 95% CI 0.34 to 2.38) and MARS (HR=1.59, 95% CI 0.73 to 3.50) had no effect on the overall survival time of MM patients (both P>0.05). Real-time RT-PCR and Western blotting showed that VARS, MARS and NARS were highly expressed in CD138+ MM cells and MM cell lines of clinical patients (all P<0.05). Cell counting and flow cytometry results showed that the proliferation of MM cells by knockout VARS was significantly inhibited (P<0.01), the proportion of apoptosis was significantly increased (P<0.05). Bioinformatics analysis showed that in addition to several pathways including the cell cycle regulated by VARS, the valine, leucine and isoleucine catabolic pathways were upregulated. Non-targeted metabolomics data showed reduced valine content in CD138+ tumor cells in MM patients compared to HD (P<0.05). HPLC results showed that compared with the scramble group, the intracellular and medium supernatant content of ARP1 cells and the medium supernatant of OCI-MY5 in the VARS-shRNA group was increased (all P<0.05). MM patients with abnormal high expression of VARS have a poor prognosis. VARS promotes the malignant growth of MM cells by affecting the regulation of valine metabolism.

Comparative volatiles profiling in milk-flavored white tea and traditional white tea Shoumei via HS-SPME-GC-TOFMS and OAV analyses.

White tea is a mildly fermented tea processed with withering and drying. Milk-flavored white tea has a unique milk flavor compared to the traditional white tea. Little is known about the aromas that make white tea taste milky. Here we conducted the volatile profiling via headspace solid-phase microextraction (HS-SPME)-gas chromatography-time-of-flight mass spectrometry (GC-TOFMS) and chemometrics to explore the key volatiles making milk-flavored white tea taste milky. Sixty-seven volatiles were identified, with 7 volatiles (OAV>1 and VIP>1) were characterized as the typical aromas. Green and light fruity scent volatiles, such as methyl salicylate, benzyl alcohol, and phenylethyl alcohol, were richer in TFs than MFs. Strong fruity and cheese aromas, such as dihydro-5-pentyl-2(3H)-furanone, 2-pentyl-furan, (E)-6,10-dimethyl-5,9-undecadien-2-one, and hexanal, were more abundant in MFs than TFs. Dihydro-5-pentyl-2(3H)-furanone, recognized as coconut and creamy aroma, should be the essential volatile for milky flavor. Also, (E)-6,10-dimethyl-5,9-undecadien-2-one and 2-pentyl-furan may contribute to the milk scent formation.

Metabolites changes of a low-temperature and low-salt fermented Chinese kohlrabi during fermentation based on non-targeted metabolomic analysis

A low-temperature and low-salt industrially fermented Chinese kohlrabi (LSCK) was developed in this study, with the salt usage decreased by approximately 70% compared to the traditional high-salt fermented Chinese kohlrabi (HSCK). The differences in physicochemical properties, metabolites and overall flavors during LSCK fermented for 0, 45 and 90 days (d) were analyzed by gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS), electronic nose (E-nose) and other techniques. The results showed that the total acid content increased significantly from 3.68 to 8.59 g/kg. However, the protein content significantly decreased from 2.52/100 to 0.66 g/100 g. The number of lactic acid bacteria cells increased significantly from 3.69 to 4.46 log10CFU/g. Based on multivariate statistical analysis, 21, 14, and 15 differential metabolites were identified in the three treatment groups A1 (0 and 45 days), A2 (45 and 90 days), and A3 (0 and 90 days) respectively (VIP &amp;gt; 1, p &amp;lt; 0.05, |log2FC| ≥ 1.1). Carbohydrates, sugar alcohols, amino acids and their derivatives were the main differential metabolites in the LSCKs fermented for different periods. Aminoacyl−tRNA biosynthesis and glycine, serine and threonine metabolism pathways significantly correlated with the differential metabolites based on Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis (p &amp;lt; 0.05). Furthermore, the overall odors were significantly different among the LSCKs with different fermentation periods, as detected by E-nose. The present study describes the change trend of metabolites during LSCK fermentation and elucidates important metabolic pathways in LSCK, providing a theoretical basis for the target regulation of functional metabolites in kohlrabi and the optimization of LSCK processing.

Impact of Light and Dark Treatment on Phenylpropanoid Pathway Genes, Primary and Secondary Metabolites in Agastache rugosa Transgenic Hairy Root Cultures by Overexpressing Arabidopsis Transcription Factor AtMYB12.

Agastache rugosa, otherwise called Korean mint, has a wide range of medicinal benefits. In addition, it is a rich source of several medicinally valuable compounds such as acacetin, tilianin, and some phenolic compounds. The present study aimed to investigate how the Tartary buckwheat transcription factor AtMYB12 increased the primary and secondary metabolites in Korean mint hairy roots cultured under light and dark conditions. A total of 50 metabolites were detected by using high-performance liquid chromatography (HPLC) and gas chromatography-time-of-flight mass spectrometry (GC-TOFMS). The result showed that the AtMYB12 transcription factor upregulated the phenylpropanoid biosynthesis pathway genes, which leads to the highest accumulation of primary and secondary metabolites in the AtMYB12-overexpressing hairy root lines (transgenic) than that of the GUS-overexpressing hairy root line (control) when grown under the light and dark conditions. However, when the transgenic hairy root lines were grown under dark conditions, the phenolic and flavone content was not significantly different from that of the control hairy root lines. Similarly, the heat map and hierarchical clustering analysis (HCA) result showed that most of the metabolites were significantly abundant in the transgenic hairy root cultures grown under light conditions. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) showed that the identified metabolites were separated far based on the primary and secondary metabolite contents present in the control and transgenic hairy root lines grown under light and dark conditions. Metabolic pathway analysis of the detected metabolites showed 54 pathways were identified, among these 30 were found to be affected. From these results, the AtMYB12 transcription factor activity might be light-responsive in the transgenic hairy root cultures, triggering the activation of the primary and secondary metabolic pathways in Korean mint.

Effect of Different Accumulative Temperate Zones in Heilongjiang on Glycine Soja Metabolites as Analyzed by Non-Target Metabolomics.

To study the effect of growth temperature on the nutritional components and metabolites of the wild soybean (Glycine soja), we analyzed the nutritional components and metabolic gases of the wild soybean in six accumulated temperature regions of the Heilongjiang Province, China, by gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS). A total of 430 metabolites, including organic acids, organic oxides, and lipids, were identified and analyzed using multivariate statistical analysis, orthogonal partial least squares discriminant analysis, principal component analysis, and cluster analysis. Eighty-seven metabolites significantly differed in the sixth accumulated temperature region compared with the other five accumulated temperature regions. The 40 metabolites (such as threonine (Thr) and lysine (Lys)) were found to be elevated in soybeans from the sixth accumulated temperature zone compared with the other five accumulated temperature zones. Through analyzing the metabolic pathways of these metabolites, amino acid metabolism had the greatest influence on wild soybean quality. The results of the amino acid analysis were consistent with those of the GC-TOF-MS and showed that amino acids in wild soybeans from the sixth accumulated temperature zone significantly differed from those of the other zones. Threonine and lysine were the main substances driving these differences. The growth temperature affected the type and concentrations of metabolites in wild soybeans, and the GC-TOF-MS analysis of the effect of growth temperature on wild soybean metabolites was shown to be feasible.

Metabolic Profiling of Primary and Secondary Metabolites in Kohlrabi (Brassica oleracea var. gongylodes) Sprouts Exposed to Different Light-Emitting Diodes.

Light-emitting diode (LED) technology is one of the most important light sources in the plant industry for enhancing growth and specific metabolites in plants. In this study, we analyzed the growth, primary and secondary metabolites of 10 days old kohlrabi (Brassica oleracea var. gongylodes) sprouts exposed to different LED light conditions. The results showed that the highest fresh weight was achieved under red LED light, whereas the highest shoot and root lengths were recorded below the blue LED light. Furthermore, high-performance liquid chromatography (HPLC) analysis revealed the presence of 13 phenylpropanoid compounds, 8 glucosinolates (GSLs), and 5 different carotenoids. The phenylpropanoid and GSL contents were highest under blue LED light. In contrast, the carotenoid content was found to be maximum beneath white LED light. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) of the 71 identified metabolites using HPLC and gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS) showed a clear separation, indicating that different LEDs exhibited variation in the accumulation of primary and secondary metabolites. A heat map and hierarchical clustering analysis revealed that blue LED light accumulated the highest amount of primary and secondary metabolites. Overall, our results demonstrate that exposure of kohlrabi sprouts to blue LED light is the most suitable condition for the highest growth and is effective in increasing the phenylpropanoid and GSL content, whereas white light might be used to enhance carotenoid compounds in kohlrabi sprouts.

In Vitro Pharmacological Activity, and Comparison GC-ToF-MS Profiling of Extracts from Cissus cornifolia (Baker) Planch.

Cissus cornifolia (Baker) Planch is traditionally used in South African traditional medicine (SATM) to treat a variety of human infections. The antimicrobial activity of extracts from C. cornifolia was investigated in vitro against a plethora of pathogenic microorganisms using the microdilution assay. The acetone extract exhibited a notable minimum inhibitory concentration (MIC) value of 0.10 mg/mL against Mycoplasma hominis and a further MIC of 0.20 mg/mL against Candida parapsilosis, Streptococcus agalactiae, Pseudomonas aeruginosa, and Enterococcus faecalis. In the antiproliferative assays, both the ethyl acetate and methanol extracts exhibited a potent inhibition of the MCF-7-21 cell line. In the anti-inflammatory assays, both the ethyl acetate and methanol extracts exhibited IC50 values of 15.59 and 15.78 µg/mL against Cyclooxygenase-2 (COX-2), respectively. Methanol extract further exhibited potent dual inhibition of both COX-2 and 15-LOX enzymes, hence, recommended to curb both related cancers, particularly breast cancer and inflammation-borne diseases. In the comparative gas chromatography time-of-flight mass spectrometry (GC/TOF-MS), the acetone, ethyl acetate, and methanol extract contained significantly prevalent amounts of compound 2-(2',4',4',6',6',8',8'-Heptamethyltetrasiloxan-2'-yloxy)-2,4,4,6,6,8,8,10,10-nonamethylcyclopentasiloxane with % area ranging from 15.714 to 39.225. The findings in the current work validates the use of the plant species in SATM in the treatment of cancer-like infections, opportunistic infections associated with HIV-AIDS. Furthermore, the in vivo studies and the mechanisms of action still need to be explored.

Fecal Volatile Metabolomics Predict Gram-Negative Late-Onset Sepsis in Preterm Infants: A Nationwide Case-Control Study.

Early detection of late-onset sepsis (LOS) in preterm infants is crucial since timely treatment initiation is a key prognostic factor. We hypothesized that fecal volatile organic compounds (VOCs), reflecting microbiota composition and function, could serve as a non-invasive biomarker for preclinical pathogen-specific LOS detection. Fecal samples and clinical data of all preterm infants (≤30 weeks' gestation) admitted at nine neonatal intensive care units in the Netherlands and Belgium were collected daily. Samples from one to three days before LOS onset were analyzed by gas chromatography-ion mobility spectrometry (GC-IMS), a technique based on pattern recognition, and gas chromatography-time of flight-mass spectrometry (GC-TOF-MS), to identify unique metabolites. Fecal VOC profiles and metabolites from infants with LOS were compared with matched controls. Samples from 121 LOS infants and 121 matched controls were analyzed using GC-IMS, and from 34 LOS infants and 34 matched controls using GC-TOF-MS. Differences in fecal VOCs were most profound one and two days preceding Escherichia coli LOS (Area Under Curve; p-value: 0.73; p = 0.02, 0.83; p < 0.002, respectively) and two and three days before gram-negative LOS (0.81; p < 0.001, 0.85; p < 0.001, respectively). GC-TOF-MS identified pathogen-specific discriminative metabolites for LOS. This study underlines the potential for VOCs as a non-invasive preclinical diagnostic LOS biomarker.