Yin Zhu

Nontargeted Analysis Using Ultraperformance Liquid Chromatography–Quadrupole Time-of-Flight Mass Spectrometry Uncovers the Effects of Harvest Season on the Metabolites and Taste Quality of Tea (Camellia sinensis L.)

The chemical composition and taste quality of tea fluctuate seasonally. However, the compounds responsible for the seasonal variation of metabolic pattern and taste quality are far from clear. This study compared the metabolite profiles of green teas of nine varieties that were plucked in spring, summer, and autumn by using ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) on a reversed phase column. A multivariate analysis indicated distinct differences among the metabolite phenotypes of teas harvested in different seasons. Heat-map analysis and metabolic pathway analysis demonstrated that flavan-3-ols, theasinensins, procyanidins, quercetin-O-glycosides, apigenin-C-glycosides, and amino acids exhibited sharp seasonal fluctuations. An equivalent quantification of tea tastes showed that in summer and autumn teas, the bitterness and astringency were significantly elevated, whereas umami declined. Metabolite content comparisons and partial least-squares analysis suggested that several flavonoids and amino acids are mainly responsible for the seasonal variations in taste quality.

Characterization of white tea metabolome: Comparison against green and black tea by a nontargeted metabolomics approach

White tea is considered the least processed form of tea and is reported to have a series of potent bioactivities, such as antioxidant, anti-inflammatory, anti-mutagenic, and anti-cancer activities. However, the chemical composition of white tea and the dynamic changes of the metabolites during the manufacturing process are far from clear. In this study, we applied a nontargeted metabolomics approach based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) to comprehensively profile the characteristic metabolites of white tea. There were significant differences in the content of amino acids, catechins, dimeric catechins, flavonol and flavone glycosides, and aroma precursors in white tea compared with green and black teas that were manufactured from the same fresh tea leaves. Furthermore, the dynamic changes of the metabolites in the tea samples with various withering durations of 0, 4, 8, 12, 16, 20, 24, 28, and 36 h were also profiled. This study offers a comprehensive characterization of the metabolites and their changes in white tea.

Enantiomeric and quantitative analysis of volatile terpenoids in different teas (Camellia sinensis)

Volatile terpenoids play important roles in the formation of tea aroma quality due to their pleasant scents and low odor thresholds. Most volatile terpenoids contain stereogenic centers, which results in various stereo distributions of their enantiomers and diastereoisomers in different types of tea. However, the distribution characteristics of terpenoid enantiomers in teas were still unclear, which poses an obstacle to the scientific understanding of tea aroma. In this work, a new and efficient analysis approach based on headspace solid phase microextraction (HS-SPME)-chiral gas chromatography-mass spectrometry (GC-MS) was established to analyze 12 pairs of familiar terpenoid enantiomers in different teas. The extraction efficiency of the HS-SPME method to extract volatile terpenoids in teas was the greatest when using CAR-DVB-PDMS (50/30μm) fibers and 1:10 proportions between tea and boiling water at a 50°C extraction temperature for 40min, and the stability observation of enantiomeric ratios of the terpenoids well proved the feasibility of the extraction method. The favorable limits of detection, limits of quantitation, repeatability, linearity, and concentration ranges of each terpenoid enantiomer demonstrated the repeatability and reliability of the analytical approach. The enantiomeric and quantitative analyses indicated that S-limonene, S-linalool, (2S, 5S)-linalool oxide A, (2S, 5R)-linalool oxide B, R-4-terpineol, (2S, 5R)-linalool oxide C, (2S, 5S)-linalool oxide D, S-α-terpineol, R-α-ionone, peak 1 of theaspirane A and peak 2 of theaspirane B were the major terpenoid components in most Chinese teas; instead, higher proportions of the opposite enantiomers of the above terpenoids were frequently detected in black teas with large leaf origin and Indonesia white teas. Besides, great diversities of enantiomeric ratios and concentrations among different teas were observed. Furthermore, partial least-squares discriminant analyses were performed to distinguish the concentration differences of the terpenoid enantiomers among different teas; the analysis results indicated that highly significant concentration differences existed between large and small leaf origins of black teas, and significant differences of the concentrations of linalool oxides A-C were observed between green, white and dark teas. The successful application of this chiral analysis technique of tea aroma will lay a scientific foundation for further quality assessment, botanical origin determination and authenticity assessment of teas.

Metabolomic analysis reveals the composition differences in 13 Chinese tea cultivars of different manufacturing suitabilities

BACKGROUND Green tea and black tea are manufactured using appropriate tea cultivars in China. However, the metabolite differences relating to the manufacturing suitability of tea cultivars are unclear. In the present study, we performed a non-targeted metabolomic analysis on 13 Chinese tea cultivars using ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry to investigate comprehensively the metabolite differences between cultivars suitable for manufacturing green tea (GT cultivars) and cultivars suitable for manufacturing both green tea and black tea (G&BT cultivars). RESULTS Multivariate statistical analysis and cluster analysis divided the 13 cultivars into two groups, namely GT cultivars and G&BT cultivars, which correlated with their manufacturing suitability. The GT cultivars contained higher levels of flavonoid glycosides, whereas the G&BT cultivars contained higher levels of catechins, dimeric catechins, phenolic acids and alkaloids. CONCLUSION Metabolic pathway analysis revealed that the flavonoid pathway inclined toward the synthesis of flavonoid glycosides in GT cultivars, whereas it inclined toward the synthesis of catechins and phenolic acids in G&BT cultivars. The results of the present study will be helpful for discriminating the manufacturing suitability of tea cultivars and investigating their breeding.

Application of metabolomics profiling in the analysis of metabolites and taste quality in different subtypes of white tea

Three subtypes of white tea, Silver Needle (SN), White Peony (WP), and Shou Mei (SM), differ in their taste, aroma, bioactivity, and commercial value. Here, a metabolomics investigation on the chemical compositions combining taste equivalent-quantification and dose-over-threshold (DoT) determination on the taste qualities were applied to comprehensively characterize the white tea subtypes for the first time. Significant differences in the contents of catechins, dimeric catechins, amino acids, phenolic acids, flavonol/flavone glycosides, and aroma precursors were observed among these 3 white teas. Metabolite content comparison and partial least-squares (PLS) analysis suggest that theanine, aspartic acid, asparagine, and AMP were positively correlated with the umami taste in white tea, and flavan-3-ols, theasinensins, procyanidin B3, and theobromine had positive correlations with higher bitterness and astringency tastes. In addition, puckering astringent (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and theogallin, bitter-tasting caffeine, and the mouth-drying/velvety-like astringent γ-aminobutyric acid (GABA) were identified as key taste compounds of white tea infusion by absolute quantification and DoT factor calculations. This work provided systematic and comprehensive knowledge on the chemical components, taste qualities, and sensory active metabolites for the subtypes of white tea.

Identification of key odorants responsible for chestnut-like aroma quality of green teas

A chestnut-like aroma is widely considered an important indicator of an excellent-quality green tea; however, the key odorants responsible for chestnut-like aroma have never been systematically studied and remain unknown. In this study, the aroma components of green teas and Chinese chestnuts were analyzed using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOFMS), and 58 compounds were identified as common aroma components among green teas, boiled Chinese chestnuts, roasted Chinese chestnuts and raw Chinese chestnuts. Subsequently, 17 volatiles, including 3-methylbutanal, (E)-3-penten-2-one, ethylbenzene, heptanal, benzaldehyde, 2-pentylfuran, octanal, benzeneacetaldehyde, (E)-2-octenal, (E,E)-3,5-octadien-2-one, linalool, nonanal, (E)-2-nonenal, decanal, (Z)-hex-3-en-1-yl hexanoate, trans-β-ionone and (E)-nerolidol, were identified as the key odorants responsible for chestnut-like aroma based on the odor activity value (OAV) calculation method. Besides, the comparison of OAVs of key odorants between fresh tea leaves and finished teas indicated that all key odorants were present in fresh tea leaves and that their contents increased or decreased during tea processing. Moreover, the comparison between results of OAV and gas chromatography-olfactometry (GC-O) methods showed that ethylbenzene, heptanal, benzaldehyde, 2-pentylfuran, (E,E)-3,5-octadien-2-one, linalool, (Z)-hex-3-en-1-yl hexanoate and trans-β-ionone were the common identified compounds between the two methods. The identification of chestnut-like aroma in green teas will provide a theoretical basis for further research on the directional adjustment and control of tea aroma quality.

Aroma formation and dynamic changes during white tea processing

The formation of and dynamic changes in aroma during white tea processing have not previously been systematically investigated. In this study, advanced comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry was employed to investigate the mechanism of white tea aroma formation. A total of 172 volatiles were identified and mainly comprising endogenous volatiles, which displayed diverse change trends during the withering period. In this process, free aroma precursor amino acids and glycosidically bound volatiles (GBVs) were found to contribute to the formation of white tea aroma, with the differential expression of aroma-related key genes accounting for various accumulation of endogenous volatiles and GBVs. In addition, the drying was also shown to play an important role in the formation of white tea aroma. Our study provides the first characterization of white tea aroma formation and establishes a theoretical basis for quality control during white tea processing operations.

Metabolomics Investigation Reveals That 8-C N-Ethyl-2-pyrrolidinone-Substituted Flavan-3-ols Are Potential Marker Compounds of Stored White Teas

White teas of different stored ages have varied flavor, bioactivity, and commercial value. In this study, a liquid chromatography-mass spectrometry-based metabolomics investigation revealed that there are distinct differences among the compound patterns of Baihaoyinzhen (BHYZ) and Baimudan (BMD) white teas with various storage durations. The levels of flavan-3-ols, procyanidins, theasinensins, theaflavins, flavonol- O-glycosides, flavone- C-glycosides, and most of the amino acids were reduced after long-term (>4 years) storage. More importantly, 8-C N-ethyl-2-pyrrolidinone-substituted flavan-3-ols (EPSFs), including seven novel compounds discovered in white teas for the first time, were formed from theanine and flavan-3-ols during storage, and their contents were positively correlated with the storage duration. These findings were further confirmed by the linearly increasing formation of EPSFs in reaction solution and BMD white teas stored in an environment-controlled cabinet. In conclusion, EPSFs were detected in white teas for the first time and were discovered as marker compounds and potential indicators for long-term storage of white tea.

The enantiomeric distributions of volatile constituents in different tea cultivars

Although the enantiomeric distribution of chiral volatiles presents great potential in discrimination of tea cultivars and their geographic origins, this area has received little attention. Thus, we herein aimed to determine the relationships between tea cultivars and the enantiomeric distributions of their chiral volatile constituents. Headspace solid-phase microextraction (HS-SPME) and enantioselective gas chromatography-mass spectrometry (Es-GC-MS) were employed to quantify 15 volatile components in 22 tea cultivars from different locations within China. The tea cultivars were successfully differentiated by their geographical origins, and the concentrations of R-linalool, S-citronellol, S-E-nerolidol, (1R, 2R)-methyl jasmonate, S-α-ionone, and the two enantiomers of linalool oxide A differed significantly among the different groups. It should also be noted that tea processing methods greatly influenced the formation of volatile enantiomers. Our results demonstrated that the enantiomeric distribution of volatile constituents closely correlates with the geographical origins, leaf types, and manufacturing suitabilities of the tea cultivars examined herein.