测定茶叶中非挥发品质代谢物的方法

Long-term storage of Liupao tea (LPT) is usually believed to enhance its quality and commercial value. The non-volatile metabolites variations and the fungal succession play a key role for organoleptic qualities during the storage procedure. To gain in-depth understanding the impact of storage time on the quality of LPT, two different brands of LPT with different storage time, including Maosheng LPTs (MS) with 0, 5, 10 and 15 years and Tianyu LPTs (TY) with 0, 3, 5, 8 and 10 years, were resorted to investigate the changes of non-volatile metabolites and fungi as well as their correlation by multi-omics. A total of 154 and 119 differential metabolites were identified in these two different brands of MS and TY, respectively, with the aid of high-performance liquid chromatography with quadrupole-time-of-flight mass spectrometry. In both categories of LPTs, the transformation of differential metabolites in the various stages referred to the formation of alkaloids, increase of organic acids, biosynthesis of terpenoids as well as glycosylation and methylation of flavonoids. Thereinto, glycosylation and methylation of flavonoids were the critical stages for distinguishing MS and TY, which were discovered in MS and TY stored for about 10 and 8 years, respectively. Moreover, the results of high-throughput sequencing showed that the key fungal genera in the storage of LPTs consisted of Eurotium, Aspergillus, Blastobotrys, Talaromyces, Thermomyces and Trichomonascus. It was confirmed on the basis of multivariate analysis that the specific fungal genera promoted the transformation of metabolites, affecting the tea quality to some extent. Therefore, this study provided a theoretical basis for the process optimization of LPT storage.

High-performance liquid chromatography (HPLC), headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and ultra-high performance liquid chromatography-Q-Exactive HF-X mass spectrometer (UHPLC-Q-Exactive HF/MS) were carried out to reveal dynamic changes of volatile and non-volatile compounds during the withering process of black tea. A total of 118 volatile organic compounds (VOCs) and 648 metabolites were identified in fresh and withered tea-leaves, respectively. Among them, 47 VOCs (OAV > 1.0) for the aroma formation, and 46 characteristic metabolites (VIP > 1.50, p < 0.01) selected through orthonormal partial least squares-discriminant analysis, indicated the withering contribution during black tea processing. Overall, the withering promoted alcohols, aldehydes, phenols, heterocyclic oxygen, hydrocarbons and halogenated hydrocarbons through relevant hydrolyzation, decomposition, terpene synthesis, and O-methylation. The hydrolyzation, O-methylation, condensation and N-acylation of kaempferol glycosides, quercetin glycosides, ester catechins, and gallic acid generated the accumulation of methoxyl flavonoids and flavonoid glucosides, dihydrokaempferol, syringic acid, theaflavins, and N-acylated amino acids, respectively.

Characteristic metabolites including tea polyphenols, amino acids, catechins, caffeine, sugars and anthocyanins were fully analyzed by high performance liquid chromatography (HPLC), gas chromatography tandem mass spectrometry (GC-MS) and ultra-high performance liquid chromatography (UHPLC)-ESI-tandem mass spectrometry (MS/MS), and showed significant differences among Zijuan tea from different plantations in Yunnan province (YN-ZJ), Qijiang (QJ-ZJ) and Ersheng (ES-ZJ) district, China, indicating that Zijuan is significantly influenced by growth conditions. Monosaccharides were the most abundant soluble sugars in YN-ZJ and ES-ZJ, while disaccharides was abundant in QJ-ZJ. d-galactose, d-mannose, d-sorbitol, inositol, d-glucose, d-galacturonic acid and raffinose involved in galactose metabolism were significantly changed (P < 0.05). Delphinidin, cyanidin, pelargonidin and their glycoside derivatives were the major anthocyanins, and showed significant differences among Zijuan samples. Flavonoids and procyanidins abundant in Zijuan provided more substrates for anthocyanins accumulation. This study presented comprehensive chemical profiling and characterized metabolites of Zijuan in different tea plantations.

Rougui Wuyi Rock tea (WRT) with special flavor can be affected by multiple factors that are closely related to the culturing regions of tea plants. The present research adopted non-targeted metabolomics based on liquid chromatography–mass spectrometry (LC-MS) and gas chromatography–mass spectrometry (GC-MS), aroma activity value method (OAV), and chemometrics to analyze the characteristic metabolites of three Rougui WRTs from different culturing regions. The results of sensory evaluation showed that the three Rougui Wuyi Rock teas had significantly different flavor qualities, especially in taste and aroma. Rougui (RG) had a heavy and mellow taste, while cinnamon-like odor Rougui (GPRG) and floral and fruity odor Rougui (HGRG) had a thick, sweet, and fresh taste. The cinnamon-like odor was more obvious and persistent in GPRG than in RG and HGRG. HGRG had floral and fruity characteristics such as clean and lasting, gentle, and heavy, which was more obvious than in RG and GPRG. The results of principal component analysis (PCA) showed that there were significant metabolic differences among the three Rougui WRTs. According to the projection value of variable importance (VIP) of the partial least squares discriminant analysis (PLS–DA), 24 differential non-volatile metabolites were identified. The PLSR analysis results showed that rutin, silibinin, arginine, lysine, dihydrocapsaicin, etc. may be the characteristic non-volatiles that form the different taste outlines of Rougui WRT. A total of 90 volatiles, including aldehydes, alcohols, esters, and hydrocarbons, were identified from the three flavors of Rougui WRT by using GC-MS. Based on OAV values and PLS-DA analysis, a total of 16 characteristic volatiles were identified. The PLSR analysis results showed that 1-penten-3-ol, α-pinene, 2-carene, β-Pinene, dehydrolinalool, adipaldehyde, D-limonene, saffron aldehyde, and 6-methyl-5-hepten-2-one may be the characteristic volatiles that form the different aroma profile of Rougui WRT. These results provide the theoretical basis for understanding the characteristic metabolites that contribute to the distinctive flavors of Rougui WRT.

… employed to investigate the metabolite profiles in tea leaves. The approach … HPLC, and GC–MS techniques, our study provided evidence that dark can influence metabolite levels in tea …

Oolong tea is a partially fermented tea with distinct tastes and aromas. However, the dynamic biochemical changes during oolong tea processing are not well understood. In this study, we performed metabolomics-based profiling of non-volatile and volatile constituents of oolong tea during its entire processing procedures by UPLC-QTOF MS and GC-TOF MS. A step-wise change of tea metabolome was observed, where catechins and oxidized products, flavonol glycosides and amino acids were identified as key discriminate metabolites. The ZuoQing process comprising alternating YaoQing and TanQing steps was deemed most critical for key metabolic transformation. Extensive YaoQing facilitated the oxidative polymerizations of catechins into theaflavins and proanthocyanidins, lowering the astringency in raw tea. Two direct terpene precursors farnesyl pyrophosphate and geranyl pyrophosphate accumulated to high levels during ZuoQing, which provided more substrates for the synthesis of downstream volatile terpenes. Moreover, both YaoQing and prolonged TanQing facilitated the formation of terpenes as well as fatty acid and benzenoid-derived volatiles, which contributed to the fruity and floral fragrances in oolong tea. The fixation step not only converted amino acids into aromatic compounds, but also lowered the amounts of flavonol glycosides, potentially improving the flavor quality of the final tea product. This study provides a comprehensive profile of flavor-related metabolic changes during oolong tea processing and will contribute to better quality control and flavor improvement of oolong tea.

The processing of tea leaves plays a crucial role in the formation of the taste of the resulting tea. In order to study the compositions of and changes in taste-related substances during the processing of Rizhao green tea, non-targeted metabolomics was used, based on UHPLC-Q Exactive MS. Totals of 529, 349, and 206 non-volatile metabolites were identified using three different detection modes, of which 112 secondary metabolites were significantly changed. Significant variations in secondary metabolites were observed during processing, especially during the drying stage, and the conversion intensity levels of non-volatile metabolites were consistent with the law of “Drying > Fixation > Rolling”. The DOT method was used to screen tea-quality-related compounds that contributed significantly to the taste of Rizhao green tea, including (−)-epicatechin gallate, (−)-epicatechin gallate, gallic acid, L-theanine, and L-leucine, which make important contributions to taste profiles, such as umami and bitterness. Metabolic pathway analysis revealed that purine metabolism, caffeine metabolism, and tyrosine metabolism perform key roles in the processing of Rizhao green tea in different processing stages. The results of this study provide a theoretical basis for tea processing and practical advice for the food industry.

… non-volatile metabolites in black tea during processing are poorly understood. In this study, the volatile and non-volatile compounds during black tea … -Exactive/MS analysis. Volatile and …

In this study, we produced roasted, baked, steamed, and sun-dried green tea products using the same batch of fresh tea leaves (FTL) of Longjing 43 (Camellia sinensis var. sinensis), and explored processing effects on the metabolic profiles of four types of green teas (FGTs) using the widely targeted metabolomics. Results showed that 146 differential metabolites including flavonoids, amino acids, lipids, and phenolic acids were screened among 1034 non-volatiles. In addition, nineteen differential metabolites were screened among 79 volatiles. Most of non-volatiles and volatiles metabolites changed notably in different manufacturing processes, whereas there were no significant differences (p＞0.05) in the levels of total catechins between FGTs and FTL. The transformation of metabolites was the dominant trend during green tea processing. The results contribute to a better understanding of how the manufacturing process influences green tea quality, and provide useful information for the enrichment of tea biochemistry theory.

… metabolites during processing, remain unclear. In this study, ultra-performance liquid chromatography–… analyze a widely targeted metabolome during green tea processing. In total, 527 …

Tea plant ( Camellia sinensis ) and its closely related species and varieties belong to Sect. Thea (L.) Dyer, Camellia L. There are abundant compounds in the fresh shoots of section Thea (L.) Dyer species and varieties. Their variation in different tea species and varieties is unclear. Fresh shoots from 336 accessions of C. sinensis and its closely related species and varieties were harvested and their non-volatile metabolites were detected through UPLC-MS (ultra-performance liquid chromatography - mass spectrometry). A total of 374 non-volatile metabolites were identified, which can be divided into 27 categories. Among them, 32 compounds were flavonoid polymers. The tea plants were divided into two groups, according to the Calinski criterion according to the composition of metabolites. The top 30 differential metabolites in C. sinensis var. sinensis , C. sinensis var. assamica , C. sinensis var. pubilimba , C. tachangensis , and C. taliensis , belong to amino acids and their derivatives, benzoic acid derivatives, carbohydrates, coumarins, flavonol glycosides, organic acids, quinoline acid and its derivatives. The results provide new insights for further understanding the characteristic metabolites of tea plant and its closely related species and varieties. non-volatile metabolites from plant ( )

… ultra-high performance liquid chromatography coupled with … profile the variations of metabolites in tea samples with various … of metabolite changes during the tea fermentation process. …

… processing technology with the shaking process of oolong tea. In this study, non-targeted … -QTOF-MS) was applied to comprehensively analyze the non-volatile metabolites of SBT …

… non-volatile metabolites. By screening for differentially expressed metabolites, we found the abundance of metabolites increased greatly after fermentation, and the non-volatile …

… Here, fixated tea samples were collected at seven different time points during the green tea … NVMs investigated via ultra-performance liquid chromatography-tandem mass spectrometry …

Camellia sinensis var. assamica cultivars 'Zijuan' (ZJ, characterized by high anthocyanin content) and 'Mengku large-leaf' (LL, with high content of catechins) are widely consumed in China. Therefore, when processed into green, white, and black teas, differences in composition and biological activities should be detectable. The aim of this work was to explore these potential differences. To achieve that, in vitro bioactivity assays and metabolomics combined with correlation and ridge analyses were applied. Metabolomics revealed that the concentrations of theasinensins, anthocyanins, and amino acids in ZJ teas were higher than those in LL teas. Compared with green and white teas, black teas had higher concentrations of Amadori rearrangement products and theaflavins. Bioactivity assays showed ZJ teas had stronger bioactivity than LL teas. Catechins, procyanidins, and flavone glycosides were identified as key contributors to bioactivity differences rather than anthocyanins. These results suggested that ZJ was more suitable for making functional tea beverages.

'Ming guan'(MG), an elite albino cultivar deriving from the progeny of the traditional albino cultivar 'Bai jiguan', is a promising candidate for white tea production due to its favorable amino acid to phenol ratio. In this study, a comprehensive metabolomics analysis using ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) and headspace solid-phase microextraction-gas chromatography mass spectrometry (HS-SPME-GC-MS) were conducted to reveal the dynamic changes of non-volatile and volatile organic compounds (VOCs) throughout the withering processing of MG white tea. Meanwhile, multivariate statistical analyses were applied to screen for the characteristic components in the flavor and aroma of MG white tea. A total of 625 non-volatile metabolites and 118 VOCs were determined, of which 90 non-volatile metabolites (VIP ≥ 1, FC ≥ 2 or ≤ 0.5) were identified as key flavor components significantly changed throughout the withering process. The relative odor activity value (ROAV) analysis highlighted 22 VOCs (ROAV ≥ 1) with substantial effect on aroma formation, of which geraniol, (E)-2-hexenal, 4-methoxy-benzaldehyde and guaiacol emerging as the most key aroma constituents of MG white tea, endowing MG white tea with fruity and floral odor notes. This study offered a comprehensive investigation into metabolite changes in MG white tea, contributing valuable insights for the innovation of new white tea products utilizing albino tea plant mutants.

The sensory quality of black tea (BT) influenced by various factors, among which tree age is particularly significant. People prefer BT produced by fresh leaves from old tea trees, yet the correlation between tree age and tea quality has not been thoroughly investigated. In this study, we analyzed the quality of BT from young trees (H-JYH) and old trees (H-OJYH) using e-tongue technology and sensory evaluation. Our findings revealed that H-OJYH had stronger sweetness and sourness, richer flavor, and diminished bitter-astringency compared to H-JYH. 1231 non-volatile metabolites and 504 volatile metabolites were discovered by ultra-performance liquid chromatography (UPLC) and gas chromatography–mass spectrometry (GC–MS). L-tartaric acid and trans-citridic acid were found to contribute to increase acidity, and 7,8-dihydroxy-6-methoxycoumarin and d-fructose 6-phosphate were associated with enhanced sweetness in H-OJYH. Additionally, lower levels of octyl gallate and vanillic acid in H-OJYH contributed to the diminished bitter-astringency. β-ionone, 2-phenylethanol and phenylacetaldehyde merged as characteristic compounds of older tree BT with stronger floral and sweet aroma. Our study serves as a guideline to explore the relationship between tree age and tea quality.

In this study, metabolomics and proteomics were performed to investigate the fluctuations of non-volatile compounds and proteins in tea leaves from three tea cultivars with varying colours during withering. A total of 2798 compounds were detected, exhibiting considerable variations in amino acids, phenylpropanoids, and flavonoids. The ZH1 cultivar displayed increased levels of amino acids but decreased levels of polyphenols, which might be associated with the up-regulation of enzymes responsible for protein degradation and subsequent amino acid production, as well as the down-regulation of enzymes involved in phenylpropanoid and flavonoid biosynthesis. The FUD and ZH1 cultivars had elevated levels of flavanols and flavanol-O-glycosides, which were regulated by the upregulation of FLS. The ZJ and ZH1 cultivars displayed elevated levels of theaflavin and peroxidase. This work presents a novel investigation into the alterations of metabolites and proteins between tea cultivars during withering, and helps with the tea cultivar selection and manufacturing development.

This study aimed to analyze the quality differences of different varieties of rose tea. Ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-…

Wuyi Rock tea, well-recognized for rich flavor and long-lasting fragrance, is a premium subcategory of oolong tea mainly produced in Wuyi Mountain and nearby regions of China. The quality of tea is mainly determined by the chemical constituents in the tea leaves. However, this remains underexplored for Wuyi Rock tea cultivars. In this study, we investigated the leaf metabolite profiles of 14 major Wuyi Rock tea cultivars grown in the same producing region using UPLC-QTOF MS and UPLC-QqQ MS with data processing via principal component analysis and cluster analysis. Relative quantitation of 49 major metabolites including flavan-3-ols, proanthocyanidins, flavonol glycosides, flavone glycosides, flavonone glycosides, phenolic acid derivatives, hydrolysable tannins, alkaloids and amino acids revealed clear variations between tea cultivars. In particular, catechins, kaempferol and quercetin derivatives were key metabolites responsible for cultivar discrimination. Information on the varietal differences in the levels of bioactive/functional metabolites, such as methylated catechins, flavonol glycosides and theanine, offers valuable insights to further explore the nutritional values and sensory qualities of Wuyi Rock tea. It also provides potential markers for tea plant fingerprinting and cultivar identification.

… MS/MS fragmentation process was accomplished at normalized collision energy of ramp from 10 to 30 eV. The accurate mass and composition of the precursor and fragment ions were …

… basis of key quality-related metabolites and harness this knowledge for precision breeding. … Another study used LC-MS/MS to analyze 'Yabukita' tea, successfully identifying …

The post-fermented tea develops enhanced quality attributes with prolonged storage. In this study, we explored the dynamic changes in sensory characteristics, untargeted and targeted metabolomics, and microbial communities of Rizhao Jinhua white tea (WFB) across different storage years. Storage process reduced bitterness and astringency, while improving overall mellowness. Levels of total polyphenols, amino acids, theaflavins, and thearubigins declined significantly, whereas theabrownin reached its peak at year 5. Among 118 identified differential metabolites, flavonoids exhibited the most pronounced variations. Further targeted quantification of flavonoids revealed catechin, epicatechin, epigallocatechin, quercitrin, and isorhamnetin as key flavor determinants. This was related to glycosylation, hydrogenation, hydroxylation, and hydrolysis reactions occurring during storage. Dominant microbial genera such as Aspergillus and Pseudomonas continuously promoted flavonoids transformations during storage. The outcomes of this research support better approaches to flavor quality optimization in stored Jinhua white tea.

… 26 nitrogen metabolism-… quality-related metabolites in tea plants at different altitudes, offering a theoretical basis for understanding the eco-physiological foundation of high-mountain tea …

… tea samples to identify potential grading-related metabolites. Amino acids, lipids and organic acids predominantly determined the superior quality of green, yellow, and white teas, … tea …

Mechanization is the inevitable future of tea harvesting, but its impact on tea chemistry and quality remains uncertain. Our study examines untargeted metabolomic data from 185 oolong tea products (Tieguanyin) made from leaves harvested by hand or machine based on UPLC-QToF-MS analysis. The data revealed a minimum 50% loss for over half of the chemicals in the machine-harvested group, including catechins, theaflavin, gallic acid, chlorogenic acid, and kaempferol-3-gluocside. Integrating sensory evaluation, OPLS-DA identified the six most important metabolites as significant contributors to sensory decline caused by harvesting mechanization. Furthermore, our research validates the possibility of using DD-SIMCA modelling with untargeted metabolomic data for distinguishing handpicked from machine-harvested tea products. The model was able to achieve 93% accuracy. This study provides crucial insights into the chemical and sensory shifts during mechanization, along with tools to manage and monitor these changes.

… The results showed that all withering treatments reduced the tea polyphenol content, while … UPLC–MS/MS analysis identified a total of 334 differential metabolites compared with …

… tea (NS) and shaken black tea (S), as well as the specific effects of shaking on quality-related metabolites and gene regulation in black tea… as well as flavonoid metabolism, thereby …

To investigate the effects of grafting on non-volatile metabolites in tea, non-targeted metabolomic analyses of fresh leaves were performed on the basis of ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF/MS). One non-grafted YingHong No. 9 and four grafted tea [grafting scion YingHong No. 9 on four different rootstocks, BaiMao No. 2 (BM2), BaiYeDanCong (BY), HeiYeShuiXian (HY), and WuLingHong (WLH)] were chosen as materials. In total, 32 differential metabolites were identified, including phenolic acids, flavan-3-ols, dimeric catechins, flavonol and flavonol/flavone glycosides, etc. Partial least squares discrimination analysis and hierarchical cluster analysis showed various effects of different rootstocks on metabolites. Thereinto, rootstocks of WLH and BY showed extremely outstanding performance in up- and downregulating these metabolites, respectively. Differential metabolites were enriched into three crucial pathways, including biosynthesis of phenylpropanoids, flavonoid biosynthesis, and flavone and flavonol biosynthesis, which might influence the quality of tea. This study provides a theoretical basis for grafting-related variations of non-volatile metabolites in fresh tea leaves.

Cultivation altitude is a comprehensive environmental factor that significantly affects tea quality. To gain a deeper understanding of the effect of cultivation altitude on tea metabolites, a widely targeted metabolomic method based on ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used to analyze tea samples derived from three altitudes (86 m, 256 m, and 880 m) of two cultivars, ‘Mingke 1’ (MK) and ‘Fuyun 6’ (FY). The results showed that distinct groups of tea samples from different altitudes and cultivars were observed based on PCA. A total of 64 and 56 altitude-related differential metabolites were identified in MK and FY, respectively. Among them, 16 compounds were consistent in both cultivars and were clustered in the metabolic pathways for flavonoid (11 compounds), amino acid (3), and fatty acid (2). The content of all flavonoids and one amino acid (L-aspartic acid) gradually decreased with increasing altitude; on the contrary, the others showed an opposite trend. Furthermore, we identified 57 differential metabolites between two cultivars. Two specific compounds (8-C-hexosyl chrysoeriol O-hexoside and pelargonidin 3-O-β-D-glucoside) were exclusively found in MK, while one compound (4-hydroxybenzoic acid) was present only in FY. These findings offer insight into the metabolic responses of tea plants to different altitudes, providing further understanding on the influence of the environment on tea plants.

In this study, the taste quality difference between high (Ninghong-Jinhao tea, JH, unfolded fresh leaves) and low (Ninghong-Congou tea, CG, unfurled fresh leaves) grades of Ninghong tea (unique black tea) was analyzed from the perspective of sensory omics, non-targeted metabolomics, and chemical dose. JH was characterized by sweetness and mellowness with umami, while CG was characterized by sweetness and thickness. A total of 94 differential metabolites contribute to the quality difference between two grades. Further quantitative analysis revealed that JH exhibited a high accumulation of amino acids, catechins, and theaflavins, while CG demonstrated a high accumulation of water extract, tea polyphenols, flavonol glycosides, and saccharides. Taste activity values (TAVs) analysis revealed that the key taste components of JH and CG were catechin, epigallocatechin gallate, three theaflavins, caffeine, myrictin-3-O-glucoside, quercetin-3-O-rutinoside, quercetin-3-O-glucoside, quercetin-3-O-galactoside, kaempferol-3-O-rutinoside, kaempferol-3-O-glucoside, and gallic acid. Among the identified compounds, the TAVs of five flavonol glycosides in Ninghong tea were found to be greater than 10 for the first time. This study is helpful to understand the taste quality difference between different grades of Ninghong tea from the molecular sensory level, providing a scientific foundation for quality improvement and targeted regulation.

Albino tea cultivars are primarily used for green tea production, with limited application in other tea types. This study used two representative albino cultivars, Huangjinya (HJY) and Baiye 1 (BY1), to prepare white tea and compare them with the normal cultivar, Fudingdabai (FDDB). BY1 exhibited superior quality owing to its high amino acid content (17.13 ± 0.32 mg/g), low catechin (50.58 ± 1.41 mg/g) and caffeine (30.09 ± 0.41 mg/g) levels, and a pleasant aroma. Pathway analysis revealed significant enrichment of differential metabolites in the flavonoid biosynthesis pathway. HJY contained abundant flavonoids, suggesting enhanced bioactivity and potential health benefits. Among the twelve key aromatic volatiles, geraniol and benzeneacetaldehyde were unique to BY1, while theaspirane was specific to HJY. This work demonstrates the influence of albino cultivars on white tea quality, providing the first evidence of their suitability for processing high-quality white tea. It advances current knowledge, offering a theoretical basis for their broader utilisation.

Simple Summary Tea is a globally popular beverage, but little is known about how altitude affects its flavor and health benefits. This study investigated the chemical differences between fresh tea leaves grown at low altitudes (350 m) and high altitudes (600 m) using advanced analytical methods. We found significant metabolic differences between the two environments, identifying over 2300 compounds, including key flavor molecules like flavonoids and phenolic acids. High-altitude samples displayed a specialized metabolic signature marked by elevated levels of tannins and stress-induced flavonoids, diverging from low-altitude samples, which possessed higher lipid and total polyphenol contents. These results demonstrated that the traditional belief that “good tea is produced from high mountains” has a solid scientific basis. Our findings provide vital information for tea growers, helping them optimize cultivation methods at different elevations to produce tea with specific, enhanced flavor profiles and health properties.

Spreading serves as a pivotal process in the flavor development of green tea. In this study, the effects of spreading under five light irradiation on the volatile and non-volatile metabolites of green tea were comprehensively investigated using intelligent sensory technologies integrated with targeted and non-targeted metabolomics analyses. The incorporation of yellow light irradiation into spreading process significantly improved the overall quality of green tea. A total of 71 volatile and 112 non-volatile metabolites were identified by GC-MS/MS and UHPLC-Q-Exactive/MS, respectively. Among them, 20 key odorants with OAVs exceeding 1 were screened out. Moreover, phenylethyl alcohol, β-damascenone, β-ionone, (E, Z)-2,6-nonadienal, linalool, and phenylacetaldehyde with higher OAVs were pivotal contributors to the aroma quality under different light irradiation. Additionally, 13 non-volatile metabolites with VIP > 1.2 were recognized as key differential metabolites under different light irradiation. The results provide technical support and theoretical guidance for enhancing the processing technology of green tea.

Background As albino tea under the geographical protection of agricultural products, Zheng’an Bai tea is not only rich in amino acids, polyphenols and other beneficial components for the human body, but also its leaf color will turn green as the temperature gradually rises, thus causing changes in the quality characteristics of tea leaves. However, these changing characteristics have not yet been revealed. Methods In-depth quality analysis was carried out on the fresh leaves of Zheng’an Bai tea at four different developmental stages and four samples from the processing stage through extensive targeted metabolomics and SPME-GC-MS analysis. Results In this study, a total of 573 non-volatile metabolites were detected from the fresh leaves and processing samples of Zheng’an Bai tea, mainly including 96 flavonoids, 75 amino acids, 56 sugars and alcohols, 48 terpenoids, 46 organic acids, 44 alkaloids, and 39 polyphenols and their derivatives. In fresh leaves, the most significant differential metabolites (VIP > 1, p < 0.05) among different samples mainly include substances such as ethyl gallate, theaflavin, isovitexin and linalool, while the main differential metabolites of samples in the processing stage include alkaloids, polyphenols and flavonoids such as zarzissine, methyl L-Pyroglutamate, theaflavin 3,3’-digallate, euscaphic acid and ethyl gallate. Overall, substances such as sugars and alcohols, alkaloids and polyphenols show the greatest differences between fresh leaves and the processing process. Meanwhile, 97 kinds of volatile metabolites were detected in these samples, most of which had a higher content in the fresh leaves. Moderate spreading is conducive to the release of the aroma of tea leaves, but fixation causes a sharp decrease in the content of most volatile metabolites. Ultimately, 9 volatile substances including geraniol, linalool, nerolidol, jasmone, octanal, 1-Nonanal, heptaldehyde, methyl salicylate and 1-Octen-3-ol were identified as the key aroma components (OAV >1) of Zheng’an Bai tea. Conclusion In conclusion, this study has for the first time comprehensively revealed the quality change characteristics of fresh leaves at different developmental stages and during the processing of Zheng’an Bai tea, and provided a foundation for further process improvement.

Steamed green tea has unique characteristics that differ from other green teas. However, the alteration patterns of non-volatile metabolites during steamed green tea processing are not fully understood. In this study, a widely targeted metabolomic method was employed to explore the changes in non-volatile metabolites during steamed green tea processing. A total of 735 non-volatile compounds were identified, covering 14 subclasses. Of these, 256 compounds showed significant changes in at least one processing step. Most amino acids, main catechins, caffeine, and main sugars were excluded from the analysis. The most significant alterations were observed during steaming, followed by shaping and drying. Steaming resulted in significant increases in the levels of most amino acids and their peptides, most phenolic acids, most organic acids, and most nucleotides and their derivates, as well as some flavonoids. Steaming also resulted in significant decreases in the levels of most lipids and some flavonoids. Shaping and drying caused significant increases in the levels of some flavonoids, phenolic acids, and lipids, and significant decreases in the levels of some amino acids and their peptides, some flavonoids, and some other compounds. Our study provides a comprehensive characterization of the dynamic alterations in non-volatile metabolites during steamed green tea manufacturing.

… Flavonoids, a class of plant polyphenols derived from plant secondary metabolism with defense functions, are widely found in plants. Some characteristic flavonoid compounds, such as …

Geographical regions profoundly influence the flavor characteristics of Congou black tea (CBT). In this study, 35 CBT samples from 7 geographical regions were comprehensively characterized by integrated multiple intelligent sensory technologies and untargeted metabolomics analysis. A satisfactory discrimination was achieved through the fusion of multiple intelligent sensory technologies (R2Y = 0.918, Q2 = 0.859). A total of 104 non-volatile and 169 volatile metabolites were identified by UHPLC-HRMS and GC–MS, respectively. Of these, 45 critical differential non-volatile metabolites and 76 pivotal differential volatile metabolites were pinpointed based on variable importance in projection >1 and p < 0.05. Moreover, 52 key odorants with OAV ≥ 1 were identified, with hexanal, phenylacetaldehyde, linalool, β-cyclocitral, methyl salicylate, geraniol, α-ethylidene phenylacetaldehyde, and trans-β-ionone being recognized as the common odorants across 7 geographical regions. The results provide theoretical support for a comprehensive understanding of the effect of geographical regions on the flavor of black tea.

To explore the influence of tea trichomes on the quality of white tea, liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS), and headspace solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) were used to identify non-volatile and volatile compounds white tea without trichomes (WTwt) and pure trichomes (PT). It was found that the bitter and astringent compounds, caffeine (CAF), epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and flavonol glycosides, were mainly enriched in the WTwt, with 16.3-fold, 47.1-fold and 28.7-fold decrease in CAF and EGCG and ECG, respectively, and the content of these compounds in PT were lower than the taste thresholds. In PT, kaempferol-3-O-(p-coumaroyl)-glucoside and kaempferol-3-O-(di-p-coumaroyl)-glucoside were non-volatile marker compounds, and decanal was significant aroma contributor with rOAV = 250.86. Moreover, the compounds in trichomes mainly contributed to the fruity and floral aroma of white tea, among which benzyl alcohol, (E)-geranylacetone, decanal, dodecanal and 6-methyl-5-hepten-2-one were the crucial aroma components, which were 2.1, 1.7, 1.8, 1.4 and 2.2 times as much as the WTwt in the PT, respectively. In conclusion, trichomes can improve the quality of white tea by reducing the bitterness and astringency, increasing the umami, as well as enhancing the fruity and floral aromas.

Harvest season exerts great influence on tea quality. Herein, the variations in non-volatile flavor substances in spring and summer fresh tea leaves of four varieties were comprehensively investigated by integrating UHPLC-Q-Exactive based lipidomics and metabolomics. A total of 327 lipids and 99 metabolites were detected, among which, 221 and 58 molecules were significantly differential. The molecular species of phospholipids, glycolipids and acylglycerolipids showed most prominent and structure-dependent seasonal changes, relating to polar head, unsaturation and total acyl length. Particularly, spring tea contained higher amount in aroma precursors of highly unsaturated glycolipids and phosphatidic acids. The contents of umami-enhancing amino acids and phenolic acids, e.g., theanine, theogallin and gallotannins, were increased in spring. Besides, catechins, theaflavins, theasinensins and flavone/flavonol glycosides showed diverse changes. These phytochemical differences covered key aroma precursors, tastants and colorants, and may confer superior flavor of black tea processed using spring leaves, which was verified by sensory evaluation.

BACKGROUND Citrus flower-green tea (CT) is a scented tea processed from green tea (GT) and fresh citrus flower, which is favored by consumers due to its potential health benefits and unique citrus flavor. This study evaluated the quality of CT and revealed the mechanism of its quality formation. RESULTS The CT had a significant citrus flavor and a good antioxidant activity, and its sensory quality was superior to that of GT. Headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) analysis revealed that the scenting process resulted in a significant increase of alkenes such as β-pinene, trans-β-ocimene, α-farnesene, isoterpinolene, and γ-terpinene, as well as a significant decrease of alcohols such as α-terpineol, L-menthol, and linalool in CT in comparision with GT. Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis revealed that the levels of flavonoids (such as neohesperidin, hesperidin, tangeritin, hesperetin 5-O-glucoside, and nobiletin) and alkaloids (such as trigonelline and theobromine) in CT increased significantly after scenting process, while the levels of amino acids (such as valine and L-phenylalanine) and organic acids (such as ascorbic acid) decreased significantly. CONCLUSION These observations showed that the scenting process promoted the absorption of aroma from citrus flowers by GT and the changes in its non-volatile metabolites, leading to the formation of citrus flavor quality in CT. This article is protected by copyright. All rights reserved.

Duyun Maojian tea (DYMJ) is distinguished by its unique processing technique and "fresh, mellow, and sweet" flavor. However, its non-volatile metabolites (NVMs) and flavor profile formation remain unclear. Herein, 3024 NVMs were identified using widely targeted metabolomics during DYMJ processing, with fixation and distinctive drying steps (light-rubbing shaping and low-temperature roasting) emerging as the critical stages of metabolites transformation. Sensory evaluation and electronic tongue analysis indicated a reduction in bitterness and astringency, alongside enhanced umami and sweetness. Correlation analysis revealed 106 taste-related NVMs, including lipids (lysophosphatidylcholines), peptides (leucyl-valyl-alanine, L-phenylalanyl-l-glutamine), and flavonoid aglycones (quercetin) exhibited high correlation (|r| > 0.8, p < 0.01) with electronic tongue-measured umami, sweetness, and bitterness. Notably, fixation established the chemical foundation for DYMJ, whereas drying likely shape its flavor quality and taste profile, particularly fresh, mellow, and sweet character. These findings enhance our understanding of DYMJ flavor formation, and provide insights for optimizing green tea processing to improve flavor.

Making tea from jujube leaves changed the chemical composition and aroma composition of jujube leaves. Here, Through LC-MS, GC-IMS, and GC-MS technology, we have revealed the effect of jujube leaf processing changes on metabolites. LC-MS identified 468 non-volatile metabolites, while GC-IMS and GC-MS detected 52 and 24 volatile metabolites, respectively. 109 non-volatile metabolites exhibiting more pronounced differences were screened. Most lipids and lipid-like molecules, organic acids, amino acids, and flavonoids increased significantly after processing. GC-IMS and GC-MS analysis revealed that the contents of aldehydes and ketones were significantly increased, while esters and partial alcohols were decreased after processing into jujube leaf tea. The main flavor substances of fresh jujube leaf and jujube leaf tea were eugenol and (E) - 2-Hexenal, respectively. Furthermore, amino acids and lipids were closely linked to the formation of volatile metabolites. Our study provided new insights into the changes in metabolites of jujube leaves processed into jujube leaf tea, and had great potential for industrial application. It laid a foundation for further research on fruit tree leaf tea.

Grade and cultivar are the important factors influencing white tea quality, but their relative metabolic contributions are not fully understood. Twelve white tea samples representing four major Fujian cultivars across three grades were analyzed using UHPLC–MS-based non-volatile metabolomics, HS-SPME–GC–MS volatile profiling, and sensory correlation analysis. In total, 47 non-volatile and 21 volatile markers were associated with grade differences, while 44 non-volatile and 26 volatile markers were linked to cultivar differences. Catechins and amino acids declined as grade decreased, whereas flavonol glycosides and gallic acid increased, accompanied by stronger astringency and reduced umami and sweetness. Aroma profiles showed a similar trend, with higher-grade teas dominated by floral notes and lower-grade teas exhibiting more herbal characteristics. Dimeric catechins, oxylipins, and aroma glycosides varied among cultivars. Volatile profiles separated the cultivars into two aroma groups: Fuding Dabai and Fuding Dahao showed more floral–fruity aromas, whereas Fuan Dabai and Zhenghe Dabai exhibited stronger herbal and aged aromas. Odor activity value analysis showed that linalool, geraniol, and (E,Z)-3,6-nonadien-1-ol were among the most abundant aroma-active compounds across white tea samples. These results provide chemical evidence for distinguishing white tea by grade and cultivar, with potential relevance to quality evaluation.

In this study, a novel spatial-resolution targeted metabolomics method was developed for a single leaf on the basis of microscale sample preparation and dansylation derivatization coupled with ultraperformance liquid chromatography-tandem mass spectrometry with a spatial resolution of 1.7 × 1.7 mm2. The practicability of this method was demonstrated by providing absolute quantitative within-leaf distribution information for 56 endogenous metabolites (including 8 catechins, 2 alkaloids, theanine, 4 theaflavins, 14 flavonol/flavone and their glycosides, 21 amino acids, and 6 phenolic acids). An application of this method in a mechanically pierced tea leaf indicated that astringent catechins, quercetin, and quercetin glycosides may be involved in the tea plant response to the wounding. In conclusion, the proposed novel method offered richest information on the within-leaf distribution of metabolites in tea to date and will be greatly helpful in understanding the defensive responses of tea plants against biotic and abiotic stresses.

… Oolong tea is a semi-fermented tea that is partially oxidised … In this study, we investigated the potential of non-targeted LC–… occurring during oolong tea manufacturing in New Zealand. …

… for untargeted metabolomics analysis. Antioxidant … teas could be completely divided into green and white tea through principal component analysis, hierarchical cluster analysis …

Intercropping is often used in the tea producing areas where land resources are not so abundant, and the produced green tea is tasted more delicious through a tea-Chinese chestnut intercropping system according to the experience of indigenous farmers. The length and weight of tea leaf increase under this intercropping system and their root systems are stratified vertically and coordinate symbiosis. However, the delicacy mechanism under the intercropping is not fully understood. Green tea from the Chinese chestnut–tea intercropping system established in the 1980s ranked highest compared with a pure tea plantation from the same region. Based on the non-targeted metabolomics, 100 differential metabolites were upregulated in the tea leaves from intercropping system relative to monoculture system. Twenty-one amino acids were upregulated and three downregulated in response to the intercropping based on the targeted metabolomics; half of the upregulated amino acids had positive effects on the tea taste. Levels of allantoic acid, sugars, sugar alcohols, and oleic acid were higher and less bitter flavonoids in the intercropping system than those in monoculture system. The upregulated metabolites could promote the quality of tea and its health-beneficial health effects. Flavone and flavonol biosynthesis and phenylalanine metabolism showed the greatest difference. Numerous pathways associated with amino acid metabolism altered, suggesting that the intercropping of Chinese chestnut–tea could greatly influence amino acid metabolism in tea plants. These results enhance our understanding of the metabolic mechanisms by which tea quality is improved in the Chinese chestnut–tea intercropping system and demonstrate that there is great potential to improve tea quality at the metabolomic level by adopting such an intercropping system.

Epigallocatechin gallate (EGCG) is the most abundant polyphenol in tea. Owing to the different fermentation degrees, differences in polyphenol composition of water extracts of green tea, white tea, oolong tea, and black tea occur, and affect health value. This study revealed that the content of EGCG decreases with the increase in the degree of fermentation. In tea with a high fermentation degree, EGCG was stably present in the form of ammoniation to yield nitrogen-containing EGCG derivative (N-EGCG). The content of N-EGCG in tea was negatively correlated with the content of EGCG. Furthermore, the content of l-serine and L-threonine in tea was positively and negatively correlated with N-EGCG and EGCG levels, respectively, suggesting that they may participate in the formation of N-EGCG as nitrogen sources. This study proposes a new fermentation-induced polyphenol-amino acid synergistic mechanism, which provides a theoretical basis for the study of the biotransformation reaction mechanism of tea polyphenols.

Natural microorganisms involved in solid-state fermentation (SSF) of Pu-erh tea have a significant impact on its chemical components. Aspergillus sydowii is a fungus with a high caffeine-degrading capacity. In this work, A. sydowii was inoculated into sun-dried green tea leaves for SSF. Metabolomic analysis was carried out by using UPLC-QTOF-MS method, and caffeine and related demethylated products were determined by HPLC. The results showed that A. sydowii had a significant (P < 0.05) impact on amino acids, carbohydrates, flavonoids, and caffeine metabolism. Moreover, A. sydowii could promote the production of ketoprofen, baclofen, and tolbutamide. Along with caffeine degradation, theophylline, 3-methylxanthine, 1,7-dimethylxanthine, 1-methylxanthine, and 7-methylxanthine were increased significantly (P < 0.05) during inoculated fermentation, which showed that demethylation was the main pathway of caffeine degradation in A. sydowii secondary metabolism. The absolute quantification analysis showed that caffeine could be demethylated and converted to theophylline and 3-methylxanthine. Particularly, about 93.24% of degraded caffeine was converted to theophylline, 27.92 mg/g of theophylline was produced after fermentation. PRACTICAL APPLICATION: Aspergillus sydowii could cause caffeine degradation in Pu-erh tea solid-state fermentation and produce theophylline through the demethylation route. Using a starter strain to ferment tea leaves offers a more controllable, reproducible, and highly productive alternative for the biosynthesis of theophylline.

Abstract The contents of hydroxycinnamoylated catechins (HCCs) in tea leaves have yet to be thoroughly investigated. Herein, four HCCs, (−)-epicatechin 3-O-p-coumaroate (EC-pC), (−)-epicatechin 3-O-caffeoate (EC-C), (−)-epigallocatechin 3-O-p-coumaroate (EGC-pC), and (−)-epigallocatechin 3-O-caffeoate (EGC-C) were synthesized as standards through an improved route and then identified in leaves of 40 and quantified in 20 tea cultivars using ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS). EC-C and EC-pC were synthesized for the first time. This study found that HCCs were widely present in the two varieties of tea, Camellia sinensis var. sinensis (CSS) and var. assamica (CSA). The contents of total four HCCs and EGC-pC were higher in CSS (729–1398 mg/kg for total HCCs, 483–1052 mg/kg for EGC-pC), while EC-C was higher in CSA (65–176 mg/kg) cultivars, suggesting that EC-C and EGC-pC could be used as biochemical markers to differentiate CSS from CSA tea cultivars. Changes in HCCs contents were detected from the tea leaves collected from April to October, indicating that tea leaves harvested in August would have the highest HCCs content.

… While chemical standardization is commonly carried out by targeted analysis using gas … quantitative 1 H NMR (qHNMR) measurements are being used increasingly in the multi-targeted …

… in selecting optimal harvesting times for tea production. An LC-MS/MS method was developed and validated for phytometabolite quantification. Epigallocatechin gallate (EGCG) …

Tibetan tea is not only a national product of geographical identity, but also a traditional beverage inherits Chinese tradition. This study evaluated the metabolic profiles and biological activity in four Tibetan teas. 83 non-volatile metabolites were identified as differentially expressed metabolites, including amino acids and their derivatives, phenolic acids, flavonoids, nucleotides and their derivatives, terpenes, alkaloids, organic acids, lipids and others. CC and 131 were rich in terpenoids and lipids. MZ contained the highest contents of amino acids and their derivatives, phenolic acids and flavonoids. 26 key volatile compounds were considered as odor-active compounds. MZ showed the highest level of antioxidant and hypoglycemic activity. Statistics analysis indicated that polyphenols, flavonoids and catechins were significantly correlated (|r| ≥ 0.7, P < 0.05) with biological activities. This study indicated significant differences in the metabolic profiles of various types of Tibetan tea, which provided a clear database for quality detection of Tibetan tea.

Six grades of Huangshan Maofeng (HSMF) green tea were studied by LC-MS based metabolomics combined with sensory evaluation on bitterness, astringency and sweet aftertaste. Although there was no significant correlation (p > 0.05) between tea grades and the contents of total polyphenols and flavonoids, non-targeted metabolomics revealed that all grades of tea could be classified into two groups, group 1 (T1, T2) and group 2 (T3, 1, 2, 3). The main marker compounds responsible for distinguishing the two groups were procyanidins, flavonoid glycosides, and four hydrolysable tannins, including monogalloyl glucose, digalloyl glucose, trigalloyl glucose and galloyl-hexahydroxydiphenoyl-glucose. The Pearson correlation coefficients of these hydrolysable tannins with HSMF green tea grades were between 0.82 and 0.95. Furthermore, their Pearson correlation coefficients regarding sweet aftertaste were in the range of 0.73-0.83. This study suggested combination of metabolomics and sensory evaluation could provide an insight in searching for more potential taste-active components.

Abstract Pruning is a routine management practice in tea cultivation. Although pruning is speculated to contribute to shoot growth and development in tea plants, it is imperative to understand the molecular mechanism involved. In order to investigate this, tea plants were pruned at different levels. Analysis of shoot growth indices revealed significant increase in shoots number and weight in shoots of pruned tea plant. Auxin assay showed higher concentrations of indole-3-acetic acid in pruned samples. Metabolomic analysis identified 80 differential metabolites in shoots of pruned plants, of which indole-3-acetonitrile and menaquinone were the common metabolites in all levels of pruning. The metabolites are involved in auxin biosynthesis, as shown by protein-protein interaction analysis. The metabolites enriched major metabolic pathways such as tryptophan metabolism, vitamin digestion and absorption, biosynthesis of ubiquinone and other terpenoid-quinone, and biosynthesis of amino acids. Genes involved in auxin signalling and menaquinone synthesis were up-regulated in pruned plants. This study reports, for the first time in nature, the synthesis of menaquinone in plants. This study concludes that pruning enhances shoot growth and development through the modulation of indole-3-acetic acid via synthesis of indole-3-acetonitrile and menaquinone in shoots, a combined effect of tryptophan metabolism and other metabolic pathways. This study contributes to knowledge in molecular mechanism of shoot growth and development.

… lower than non-roasted teas. Targeted taste-compounds metabolomics revealed that (–)-… The Pearson correlation coefficient analysis suggested procyanidin B2, coumaroylquinic …

Cordyceps militaris (C. militaris) has been approved and widely used in healthy food. The present study aimed to improve the flavor of summer Keemun black tea (KBT) using C. militaris solid-state fermentation. Combined with sensory evaluation, the volatile and non-volatile components of solid-state fermentation of KBT (SSF-KBT) and KBT were analyzed. The results showed that after the solid-state fermentation, the contents of total polyphenol, total flavonoid, and total free amino acids were significantly reduced. Further non-targeted metabolomics analysis revealed that the contents of non-galloylated catechins and d-mannitol increased, while the galloylated catechins and flavonoid glycosides decreased as did the bitterness and astringency of KBT. Dihydro-β-ionone and β-ionone (OAV = 59321.97 and 8154.17) were the aroma-active compounds imparting woody and floral odors in SSF-KBT, respectively. Current study provides a new avenue to develop summer-autumn KBT.

… Theanine (N-ethyl-glutamine) is the most abundant amino acid in tea. As a non-UV-active … Theogallin is a depside of quinic and gallic acids, and a characteristic compound of tea. …

Large-leaf yellow tea (LYT) is made from mature tea leaves with stems and has unique sensory characteristics different from other teas. To study the chemical changes of LYT during processing, samples were collected from each step for quantitative and qualitative analyses by high-performance liquid chromatography and liquid chromatography-mass spectrometry (LC-MS). LC-MS-based nontargeted and targeted metabolomics analyses revealed that the tea sample after roasting was markedly different from samples before roasting, with the levels of epicatechins and free amino acids significantly decreased, but the epimerized catechins increased dramatically. After accounting for common compounds in tea, N-ethyl-2-pyrrolidinone-substituted flavan-3-ols were found to be the marker compounds responsible for the classification of all samples, as they rapidly rose with increasing processing temperature. These findings suggested that the predominant changes in the tea constituents during large-leaf yellow tea roasting were the thermally induced degradation and epimerization of catechins and the formation of N-ethyl-2-pyrrolidinone-substituted flavan-3-ols from l-theanine.

… The inhibitory effects of pickled tea samples on α-amylase and α-… The results showed that glycosides and most organic acids … the chemical variation of pickled tea during processing, and …

… the tea varieties for green tea manufacturing, … glycosides was the lowest in the tea variety for oolong tea manufacturing. The content of individual phenolic compounds in the selected tea …

Mulberry (Morus alba L.) leaves from two cultivars, Yai-Burirum (YB) and Khunphai (KP), were prepared into green tea (GT) and black tea (BT). Compared to fresh leaf (FL) extract, GT and BT extracts were evaluated for their total phenolic and total flavonoid contents. Total phenolic content (TPCs) in all samples ranged between 129.93 and 390.89 mg GAE/g extract. The processing of tea decreased the levels of TPC when compared to FL extracts in both cultivars. The total flavonoid content (TFCs) in all samples was found in the range of 10.15–39.09 mg QE/g extract and TFCs in GT and BT extracts were higher than FL extracts. The change in tryptophan, melatonin, phenolic and flavonoid contents was investigated by liquid chromatography–mass spectroscopy (LC-MS). The results exhibited that tryptophan contents in all samples were detected in the range 29.54–673.72 µg/g extract. Both GT and BT extracts increased tryptophan content compared to FL extracts. BT extracts presented the highest amounts of tryptophan among others in both cultivars. Phenolic compounds were found in mulberry leaf extracts, including gallic acid, caffeic acid, gentisic acid, protocatechuic acid and chlorogenic acid. Chlorogenic acid presented the highest amount in all samples. Almost all phenolic acids were increased in the processed tea extracts except chlorogenic acid. Rutin was the only flavonoid that was detected in all extracts in the range 109.48–1009.75 mg/g extract. The change in phenolic and flavonoid compounds during tea processing resulted in the change in antioxidant capacities of the GT and BT extracts. All extracts presented acetylcholinesterase enzyme (AChE) inhibitory activity with IC50 in the range 146.53–165.24 µg/mL. The processing of tea slightly increased the AChE inhibitory effect of GT and BT extracts. In conclusion, processed tea from mulberry leaves could serve as a new alternative functional food for health-concerned consumers because it could be a promising source of tryptophan, phenolics and flavonoids. Moreover, the tea extracts also had antioxidative and anti-AChE activities.

Polyphenols, the most abundant components in tea, determine the quality and health function of tea. The analysis of polyphenols in tea is a topic of increasing interest. However, the complexity of the tea matrix, the wide variety of teas, and the difference in determination purposes puts forward higher requirements for the detection of tea polyphenols. Many efforts have been made to provide a highly sensitive and selective analytical method for the determination and characterization of tea polyphenols. In order to provide new insight for the further development of polyphenols in tea, in the present review we summarize the recent literature for the detection of tea polyphenols from the perspectives of determining total polyphenols and individual polyphenols in tea. There are a variety of methods for the analysis of total tea polyphenols, which range from the traditional titration method, to the widely used spectrophotometry based on the color reaction of Folin–Ciocalteu, and then to the current electrochemical sensor for rapid on-site detection. Additionally, the application of improved liquid chromatography (LC) and high-resolution mass spectrometry (HRMS) were emphasized for the simultaneous determination of multiple polyphenols and the identification of novel polyphenols. Finally, a brief outline of future development trends are discussed.

Marketed green teas (GTs) can highly vary in their chemical composition, due to different origins, processing methods, and a lack of standardization of GT-based products. Consequently, biological activities become difficult to correlate to the presence/content of certain constituents. Herein, ultra-high-performance liquid chromatography (UHPLC) combined with high-resolution tandem mass spectrometry (HR MS/MS) was successfully applied to six commercial GT products, extracted by ethanol sonication, to disclose their polyphenol profile beyond the well-known catechins. The relative abundance of each class of metabolites was correlated to antiradical and antilipoperoxidant data through hierarchical clustering analysis, since it reasonably affects the beneficial properties of the product that reaches the consumer. The thiobarbituric acid reactive substances (TBARS) assay demonstrated that GT extracts effectively counteracted the UV-induced lipoperoxidation of hemp oil, which is highly rich in Polyunsaturated Fatty Acids (PUFAs), and therefore highly unstable. The Relative Antioxidant Capacity Index (RACI) comprehensively emphasized that gunpower and blend in filter GTs appeared to be the less active matrices, and except for a GT-based supplement, the Sencha GT, which was particularly rich in flavonol glycosides, was the most active, followed by Bancha GT.

… in amino acid levels between tea types and countries. … such as phenolic acids, flavonoid glycosides, catechins, … through-put and comprehensive LC–MS method utilising an alternative …

Wuyi Rock tea, specifically Shuixian and Rougui, exhibits distinct sensory characteristics. In this study, we investigated the sensory and metabolite differences between Shuixian and Rougui. Quantitative description analysis revealed that Rougui exhibited higher intensity in bitter, thick, harsh, and numb tastes, while Shuixian had stronger salty and umami tastes. Nontargeted metabolomics identified 151 compounds with 66 compounds identified as key differential metabolites responsible for metabolic discrimination. Most of the catechins and flavonoids were enriched in Rougui tea, while epigallocatechin-3,3'-di-O-gallate, epigallocatechin-3,5-di-O-gallate, gallocatechin-3,5-di-O-gallate, isovitexin, and theaflavanoside I were enriched in Shuixian tea. Catechins, kaempferol, quercetin, and myricetin derivatives were positively correlated with bitter taste and numb sensation. Sour taste was positively correlated to organic acids. Amino acids potentially contributed to salty and umami tastes. These results provide further insights into the taste characteristics and the relationship between taste attributes and specific metabolites in Wuyi Rock tea.

… C-bonded glycosides, as O-bonded glycosides are normally … tea LC–MS n data set, including C-glycosylated polyphenols (… glycosyls and quinic acids, saponins and phenol glycosides (…

The effect of fermentation process on the formation of quality compounds of crush-tear-curl black tea (CTCBT) was unclear. In this study, a total of nine characteristic volatile compounds were screened, and their contents generally exhibited an upward trend during the fermentation process. Among them, the increase in the contents of phenylethanal, 2,4-hexadienal, and guaiacol contributed to the formation of a sweet aroma in tea, while the increase in hexanal and (2E)-hexanal contents helped suppress the production of off-flavors. Additionally, 68 non-volatile differential metabolites were identified. Fermentation primarily influenced the biosynthesis of flavonoids and amino acids, and regulated the taste quality of tea by promoting the oxidative degradation of tea polyphenols, the acylation of amino acids, and the breakdown of nucleotides. This provides an in-depth understanding of the dynamic evolution of quality compounds during the fermentation of CTCBT.

… -dimensional space, the pile-fermentation process of dark green tea can be divided into four … flavor of dark green tea. This study demonstrated the fungal succession, non-volatile flavor …

The present study investigated the dynamics changes in physicochemical properties and non-volatile metabolites during Bulang pickled tea fermentation. A combination of artificial sensory evaluation, chemical-physical analysis, ultra performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS), and multivariate statistical analysis were employed to examine the differences among four fermentation stages of Bulang pickled tea. The bitterness, astringency, sweetness after taste, sourness and fermentation taste tended to increase with fermentation. The highest lactic acid bacteria, aerobic bacteria, total titratable acidity, total soluble sugar, total polyphenols, and total flavonoids were recorded at the second month of fermentation, while fungi, total free amino acids, total antioxidant capacity and hydroxyl free radical scavenging capacity increased with fermentation. Mantel test demonstrated significant associations between lactic acid bacteria /fungal communities and taste characteristics. UPLC-QTOF-MS analysis led to the identification of 35 differential non-volatile metabolites, predominantly comprising heterocyclic compounds, organic acids with their derivatives, and flavonoids. Nine non-volatile metabolites are related to antioxidant activity, and morin, malvidin and 7-methylxanthine exhibit relatively strong antioxidant activity. This study provides comprehensive insights into the non-volatile metabolites and antioxidant function of Bulang pickled tea.

… previously been employed to identify non-volatile metabolites of dark green tea during pile-… The LC/MS system for metabolomics analysis was composed of Waters Acquity I-Class PLUS …

The mechanism of the quality formation of dark tea is not fully clear, particularly under variable fermentation temperatures. In this study, the tea fermented with Aspergillus niger (AN) at 25 (AN25) and 37 °C (AN37) exhibited the highest quality. Different fermentation temperatures primarily influenced the degradation of fatty acids and the hydrolysis of glycosides in the tea, with 37 °C being the most favorable for the release and accumulation of volatile compounds. Eighteen key volatiles were identified. Among these, benzaldehyde (a 120.9% increase compared to CK), α-ionone (957.8%), linalool (172.2%), and nonanal (22.8%) were present at high levels in AN37, and these compounds served as the main aroma contributors. Inoculation with AN and fermentation temperature primarily influences the levels of total polyphenols, organic acids and their derivatives, as well as amino acids and their metabolites in dark tea. Total polyphenols, flavonoids, and nucleotide and its metabolites were more rapidly consumed at 25–37 °C, contributing to the improved taste of the tea infusion. Additionally, EGC, GC, melezitose, and sucrose showed significant negative correlations with the taste quality of the tea infusion (p < 0.05). These results are conducive to further understanding of the quality formation of dark tea.

Fermentation significantly influences the chemical composition of black tea, yet the effects of different fermentation temperatures on non-volatile components and their in vitro hypoglycemic activity are insufficiently studied. This research investigates how varying temperatures (20, 25, and 30°C) affect the bioactive profile and the inhibitory activity of Jinxuan black tea against α-glucosidase and α-amylase. Our results show that lower fermentation temperatures (20°C) lead to elevated levels of key bioactive compounds, including tea polyphenols (9.24%), soluble sugars (8.24%), thearubigins (7.17%), and theasinesin A (0.15%). These compounds correlate strongly with enhanced α-glucosidase inhibition (R = 0.76–0.97). Non-targeted metabolomic analysis revealed that 36 differential metabolites, including catechins, exhibited altered levels with increasing fermentation temperature. Notably, tea fermented at 20°C exhibited superior hypoglycemic activity, with α-glucosidase inhibition (IC50 = 14.00 ± 1.00 μg/ml) significantly outperforming α-amylase inhibition (IC50 = 2.48 ± 0.28 mg/ml). The findings of this research underscore the importance of fermentation temperature in optimizing the bioactive profile of black tea. It is proposed that recommendations for future processing or formulation should emphasize the use of lower fermentation temperatures, aimed at augmenting the health benefits linked to higher polyphenol content and stronger hypoglycemic activity.

Eurotium cristatum is a promising probiotic fungus for improving tea flavor. This research explored the volatile and non-volatile metabolites associated with flavor characteristics of E. cristatum fermented old leaves of Anji Baicha, utilizing e-nose, e-tongue, HS-SPME-GC-MS, and LC-MS. The results indicated that the reduction in bitterness and astringency post-fermentation correlated with the downregulation of catechins, chlorogenic acid, proanthocyanidins, and certain amino acids. The enhanced umami taste was primarily due to the upregulation of gallic acid, adenine, aspartic acid and glutamic acid. GC-MS analysis identified 28 pivotal aroma compounds with OAV > 1, among which β-ionone, 6-methyl-5-hepten-2-one, and 1-octen-3-ol were identified as primary contributors to the distinctive fungal floral scent. Notably, the degradation and oxidation of non-volatile metabolites facilitated the enrichment of volatile compounds. These insights deepen the understanding of metabolite-flavor profile relationship in Anji Baicha fermented by E. cristatum, offering strategies for enhancing added value of old leaves of Anji Baicha.

Pu-erh raw tea (PRT), a post-fermented tea, is prized for its complex flavor profile and health-promoting properties. While extended storage enhances its sensory attributes, the decade-scale metabolic dynamics underpinning flavor evolution remain unexplored. This study comprehensively characterized non-volatile metabolomic profiles and flavor changes in PRT across a 10-year storage period (2012-2023). Integrated sensory evaluation and electronic tongue analysis indicated progressive darkening of the infusion attributed to oxidation and increased mellowness in aged samples, though instrumental taste differentiation was limited. Untargeted metabolomics identified 3384 metabolites, with 619 consistently present across all storage years. Crucially, γ-linolenic acid and gallic acid demonstrated significant time-dependent accumulation, quantitatively validated via 1H quantitative NMR (qNMR). These metabolites emerged as robust markers for discriminating PRT storage duration, revealing novel biochemical drivers of its long-term aging characteristics.

… Local residents' storage practices for KJGT found that aged … 2228 non-volatile metabolites covering 13 categories were … in non-volatile metabolites among three different storage …

As a semifermented tea, oolong is exceedingly popular worldwide for its elegant, flowery aroma and mellow, rich taste. However, recent marketing trends for old oolong teas and their chemical quality largely remain unexplored. In this study, we applied widely targeted metabolomics using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with multivariate analysis to investigate the chemical change of oolong teas in the aging process. With the increasing of store time, most nongalloylated catechins; tannins, including TFs and proanthocyanidins; flavonols and glycosylated flavonols; amino acids and their derivatives; nucleotides and their derivatives; and lots of alkaloids and phospholipids declined, while most fatty acids and organic acids increased, and galloylated catechins, GA, and caffeine were almost stable. The result also suggested that approximately seven years (but not an infinite extension) was a special period for oolong tea storage, which brings about excellent taste.

Withering is a crucial step in the production of white tea. However, the non-volatile metabolomic profile of white tea produced from the Longjing 43 cultivar using heated withering methods has not been fully elucidated. This study investigated the effect of a novel heated withering process on the taste quality and non-volatile metabolome of white tea. The application of heat significantly shortened the withering duration and enhanced the sensory quality of the tea. Specifically, white tea withered using hot air for 23 h, sunlight for 28 h, and fluorescent lamp for 31 h, all at 28 ± 1 °C, achieved higher taste sensory evaluation scores compared to traditional room-temperature withering at 20 ± 1 °C for 40 h. Non-targeted metabolomic analysis led to the identification of 1,372 metabolites across the different withering treatments. Relative to room-temperature withering, the heated withering processes—particularly hot air withering—resulted in higher concentrations of key taste-related compounds, including organic acids, amino acids, flavonoids, and catechins. Enrichment analysis of KEGG pathways indicated that the main metabolic pathways affected by heated withering were cysteine and methionine metabolism, glutathione metabolism, and isoquinoline alkaloid biosynthesis, with the most pronounced effects observed under hot air withering. In summary, hot air withering not only reduced the processing time but also improved the taste quality and markedly altered the profile of key metabolites in white tea.

… capabilities, high sensitivity, and broad analytical range, we use UPLC-MS/MS to 90 … were collected after each processing step and stored at −80℃. Leaves at different 112 …

… and non-volatile metabolomic analyses indicated that aging primarily affects metabolite … The experimental results illustrate that the improvement in CCP quality induced by aging does …

… However, there were no studies between tree age and tea … 1231 non-volatile metabolites and 504 volatile metabolites … reveal the effects of different storage years on metabolites 499 …

Blister blight infected with Exobasidium vexans is one of the most destructive foliar diseases that seriously affects the quality and yield of tea. This research was to investigate the metabolite changes of healthy and infected leaves on tea cultivar "Fuding Dabaicha", as well as further explore the potential antimicrobial substances against E. vexans infection. In total, 1166 compounds were identified in the whole stage of infection, among which 73 different common compounds were significantly accumulated involved in the important antimicrobial substances of flavonoids and phenolic acids, including kaempferol (3,5,7,4'-tetrahydroxyflavone), kaempferol-3-O-sophoroside-7-O-glucoside, phloretin, 2,4,6-trihydroxybenzoic acid, galloylprocyanidin B4, and procyanidin C1 3'-O-gallate, which indicated that these metabolites might positively dominate the resistance to E. vexans. Furthermore, the relevant biological pathways, such as "Flavone and flavonol biosynthesis", "Flavo-noid biosynthesis", and "Phenylpropane pathway", were more closely related to the resistance against E. vexans. Additionally, total flavonoids, phenolics, alkaloids and terpenoids contributing to antimicrobial and antioxidant capacity altered significantly in four different infection periods, especially the Leaf_S2 stage (the second stage of infection) in which the concentration accumulated the most. The leaves affected by E. vexans infection at the second stage had the relatively highest antioxidant activity. Accordingly, this study provided a theoretical support and comprehensive insights into the effects on the metabolite changes, tea quality components and antioxidant activity of blister blight caused by E. vexans.

Electrospray tandem mass spectrometry (ESI-MS/MS) is commonly used in high throughput metabolomics. One of the key obstacles to the effective use of this technology is the difficulty in interpreting measured spectra to accurately and efficiently identify metabolites. Traditional methods for automated metabolite identification compare the target MS or MS/MS spectrum to the spectra in a reference database, ranking candidates based on the closeness of the match. However the limited coverage of available databases has led to an interest in computational methods for predicting reference MS/MS spectra from chemical structures. This work proposes a probabilistic generative model for the MS/MS fragmentation process, which we call Competitive Fragmentation Modeling (CFM), and a machine learning approach for learning parameters for this model from MS/MS data. We show that CFM can be used in both a MS/MS spectrum prediction task (ie, predicting the mass spectrum from a chemical structure), and in a putative metabolite identification task (ranking possible structures for a target MS/MS spectrum). In the MS/MS spectrum prediction task, CFM shows significantly improved performance when compared to a full enumeration of all peaks corresponding to substructures of the molecule. In the metabolite identification task, CFM obtains substantially better rankings for the correct candidate than existing methods (MetFrag and FingerID) on tripeptide and metabolite data, when querying PubChem or KEGG for candidate structures of similar mass.

Database search is a widely used approach for identifying metabolites from tandem mass spectra (MS/MS). In this strategy, an experimental spectrum is matched against a user-specified database of candidate metabolites, and candidates are ranked such that true metabolite-spectrum matches receive the highest scores. Machine-learning methods have been widely incorporated into database-search-based identification tools and have substantially improved performance. To further improve identification accuracy, we propose a new framework for generating negative training samples. The framework first uses autoencoders to learn latent representations of metabolite structures and MS/MS spectra, thereby recasting metabolite-spectrum matching as matching between latent vectors. It then uses a GAN to generate latent vectors of decoy metabolites and constructs decoy metabolite-spectrum matches as negative samples for training. Experimental results show that our tool, MS2MetGAN, achieves better overall performance than existing metabolite identification methods.

Here are presenting the blank based time-alignment (BBTA) as a strong analytical approach for treatment of non-linear shift in time occurring in HPLC-MS data. Need of such tool in recent large dataset produced by analytical chemistry and so-called omics studies is evident. Proposed approach is based on measurement and comparison of blank and analyzed sample evident features. In the first step of BBTA procedure, the number of compounds is reduced by max-to-mean ratio thresholding, which extensively reduce the computational time. Simple thresholding is followed by selection of time markers defined from blank inflex points which are then used for the transformation function, polynomial of second degree, in the example. BBTA approach was compared on real HPLC-MS measurement with Correlation Optimized Warping (COW) method. It was proved to have distinctively shorter computational time as well as lower level of mathematical presumptions. The BBTA is computationally much easier, quicker (more then 1000x) and accurate in comparison with warping. Moreover, markers selection works efficiently without any peak detection. It is sufficient to analyze only baseline contribution in the analyte measurement with sparse knowledge of blank behavior. Finally, BBTA does not required usage of extra internal standards and due to its simplicity it has a potential to be widespread tool in HPLC-MS data treatment.

Mass spectrometry (MS) plays a critical role in molecular identification, significantly advancing scientific discovery. However, structure elucidation from MS data remains challenging due to the scarcity of annotated spectra. While large-scale pretraining has proven effective in addressing data scarcity in other domains, applying this paradigm to mass spectrometry is hindered by the complexity and heterogeneity of raw spectral signals. To address this, we propose MS-BART, a unified modeling framework that maps mass spectra and molecular structures into a shared token vocabulary, enabling cross-modal learning through large-scale pretraining on reliably computed fingerprint-molecule datasets. Multi-task pretraining objectives further enhance MS-BART's generalization by jointly optimizing denoising and translation task. The pretrained model is subsequently transferred to experimental spectra through finetuning on fingerprint predictions generated with MIST, a pre-trained spectral inference model, thereby enhancing robustness to real-world spectral variability. While finetuning alleviates the distributional difference, MS-BART still suffers molecular hallucination and requires further alignment. We therefore introduce a chemical feedback mechanism that guides the model toward generating molecules closer to the reference structure. Extensive evaluations demonstrate that MS-BART achieves SOTA performance across 5/12 key metrics on MassSpecGym and NPLIB1 and is faster by one order of magnitude than competing diffusion-based methods, while comprehensive ablation studies systematically validate the model's effectiveness and robustness.

Untargeted metabolomic studies are revealing large numbers of naturally occurring metabolites that cannot be characterized because their chemical structures and MS/MS spectra are not available in databases. Here we present iMet, a computational tool based on experimental tandem mass spectrometry that could potentially allow the annotation of metabolites not discovered previously. iMet uses MS/MS spectra to identify metabolites structurally similar to an unknown metabolite, and gives a net atomic addition or removal that converts the known metabolite into the unknown one. We validate the algorithm with 148 metabolites, and show that for 89% of them at least one of the top four matches identified by iMet enables the proper annotation of the unknown metabolite. iMet is freely available at http://imet.seeslab.net.

We present ProbMetab, an R package which promotes substantial improvement in automatic probabilistic LC-MS based metabolome annotation. The inference engine core is based on a Bayesian model implemented to: (i) allow diverse source of experimental data and metadata to be systematically incorporated into the model with alternative ways to calculate the likelihood function and; (ii) allow sensitive selection of biologically meaningful biochemical reactions databases as Dirichlet-categorical prior distribution. Additionally, to ensure result interpretation by system biologists, we display the annotation in a network where observed mass peaks are connected if their candidate metabolites are substrate/product of known biochemical reactions. This graph can be overlaid with other graph-based analysis, such as partial correlation networks, in a visualization scheme exported to Cytoscape, with web and stand alone versions. ProbMetab was implemented in a modular fashion to fit together with established upstream (xcms, CAMERA, AStream, mzMatch.R, etc) and downstream R package tools (GeneNet, RCytoscape, DiffCorr, etc). ProbMetab, along with extensive documentation and case studies, is freely available under GNU license at: http://labpib.fmrp.usp.br/methods/probmetab/.

There are different problems for resolution of complex LC-MS or GC-MS data, such as the existence of embedded chromatographic peaks, continuum background and overlapping in mass channels for different components. These problems cause rotational ambiguity in recovered profiles calculated using multivariate curve resolution (MCR) methods. Since mass spectra are sparse in nature, sparsity has been proposed recently as a constraint in MCR methods for analyzing LC-MS data. There are different ways for implementation of the sparsity constraint, and majority of methods rely on imposing a penalty based on the L0-, L1- and L2-norms of recovered mass spectra. Ridge regression and least absolute shrinkage and selection operator (Lasso) can be used for implementation of L2- and L1-norm penalties in MCR, respectively. The main question is which Lx-norm penalty is more worthwhile for implementation of the sparsity constraint in MCR methods. In order to address this question, two and three component LC-MS data were simulated and used for the case study in this work. The areas of feasible solutions (AFS) were calculated using the grid search strategy. Calculating Lx-norms values in AFS for x between zero and two revealed that the gradient of optimization surface increased from x values equal to two to x values near zero. However, for x equal to zero, the optimization surface was similar to a plateau, which increased the risk of sticking in local minima. Generally, results in this work, recommend the use of L1-norm penalty methods like Lasso for implementation of sparsity constraint in MCR-ALS algorithm for finding more sparse solutions and reducing the extent of rotational ambiguity.

Metabolomics is a key approach in modern functional genomics and systems biology. Due to the complexity of metabolomics data, the variety of experimental designs, and the variety of existing bioinformatics tools, providing experimenters with a simple and efficient resource to conduct comprehensive and rigorous analysis of their data is of utmost importance. In 2014, we launched the Workflow4Metabolomics (W4M, http://workflow4metabolomics.org) online infrastructure for metabolomics built on the Galaxy environment, which offers user-friendly features to build and run data analysis workflows including preprocessing, statistical analysis, and annotation steps. Here we present the new W4M 3.0 release, which contains twice as many tools as the first version, and provides two features which are, to our knowledge, unique among online resources. First, data from the four major metabolomics technologies (i.e., LC-MS, FIA-MS, GC-MS, and NMR) can be analyzed on a single platform. By using three studies in human physiology, alga evolution, and animal toxicology, we demonstrate how the 40 available tools can be easily combined to address biological issues. Second, the full analysis (including the workflow, the parameter values, the input data and output results) can be referenced with a permanent digital object identifier (DOI). Publication of data analyses is of major importance for robust and reproducible science. Furthermore, the publicly shared workflows are of high-value for e-learning and training. The Workflow4Metabolomics 3.0 e-infrastructure thus not only offers a unique online environment for analysis of data from the main metabolomics technologies, but it is also the first reference repository for metabolomics workflows.

While there are many different methods for peak detection, no automatic methods for marking peak boundaries to calculate area under the curve (AUC) and signal-to-noise ratio (SNR) estimation exist. An algorithm for the automation of liquid chromatography tandem mass spectrometry (LC-MS/MS) mass chromatogram quantification was developed and validated. Continuous wavelet transformation and other digital signal processing methods were used in a multi-step procedure to calculate concentrations of six different analytes. To evaluate the performance of the algorithm, the results of the manual quantification of 446 hair samples with 6 different steroid hormones by two experts were compared to the algorithm results. The proposed approach of automating mass chromatogram quantification is reliable and valid. The algorithm returns less nondetectables than human raters. Based on signal to noise ratio, human non-detectables could be correctly classified with a diagnostic performance of AUC = 0.95. The algorithm presented here allows fast, automated, reliable, and valid computational peak detection and quantification in LC- MS/MS.