Identification of Chinese green tea (Camellia sinensis) marker metabolites using GC/MS and UPLC-QTOF/MS.

Tea is one of the most widely consumed aromatic beverages in the world because of its taste and flavor, as well as due to many potential health beneficial properties. Metabolomics focuses on an in-depth analysis of all metabolites in living organisms. In this study, 29 primary metabolites and 25 secondary metabolites were identified using GC/MS and UPLC-QTOF/MS, respectively. Further, PCA analysis showed conspicuous discrimination for the ten varieties of green tea with metabolite profiling. Among them, organic acids, amino acids, flavan-3-ols, and flavonol glycosides varied greatly through checking the VIP values of the PLS-DA model. Moreover, the intrinsic and/or extrinsic factors characterizing each type of green tea were also discussed. The chemical component marker derived here should be used as an important detection index, while evaluating the tea quality, as well as while establishing the tea quality standard. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00970-4.

Evaluation of in-vitro antimicrobial activity of Artemisia apiacea H. and Scutellaria baicalensis G. extracts.

PURPOSE: In traditional Korean medicine, Artemisia apiacea H. (ART) and Scutellaria baicalensis G. (SCU) are combined for the treatment of malaria and other malaria-like diseases. Because SCU is well-known as an antibacterial agent, the antimicrobial effect of a mixture of ART and SCU was investigated. METHODOLOGY: Plant samples were purchased from Kyungdong mart and extracted with 70 % ethanol. The in vitro antimicrobial activity of ART and SCU against pathogenic fungi (Aspergillus niger, Aspergillus oryzae, Candida albicans, Candida tropicalis and Candida glabrata), Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus) was evaluated using a broth microdilution assay, modified-disc diffusion and agar dilution methods with further CH2Cl2-fractionated ART, SCU and a mixture of ART/SCU (at a ratio of 3 : 5) (THAN-1). RESULTS: ART and SCU were effective against A. niger, C. albicans, B. subtilis and S. aureus. The range of minimum inhibitory concentration (MIC) values was 0.03125 to 4 mg ml-1 in the ART and SCU treatments. ART exhibited stronger activity than SCU. Interestingly, a 3 : 5 ratio mixture of ART and SCU (THAN-1) showed stronger antimicrobial activity than ART or SCU used individually. CONCLUSION: A treatment using a mixture of herbs such as THAN-1 would be useful in the suppression of the growth of pathogenic bacterial and fungal strains.

Enhancement of Protective Effects of Radix Scutellariae on UVB-induced Photo Damage in Human HaCaT Keratinocytes.

Radix Scutellariae (RS) has long been used in the treatment of inflammatory and allergic diseases. Its main flavonoids, baicalin (BG) and wogonoside (WG), can be hydrolyzed into their corresponding aglycones, baicalein (B) and wogonin (W). In this study, we developed a safe and effective method of transforming these glycosides using Peclyve PR. The transformation rate of BG and WG reached 98.5 and 98.1%, respectively, with 10% enzyme at 40 °C for 60 h. Furthermore, we compared the anti-photoaging activity of RS before and after enzyme treatment, as well as their respective main components, in UVB-irradiated HaCaT cells. Results found that enzyme-treated RS (ERS) appeared to be much better at preventing UVB-induced photoaging than RS. ERS significantly inhibited the upregulation of matrix metalloproteinase-1 and IL-6 caused by UVB radiation by inactivating the MAPK/AP-1 and NF-κB/IκB-α signaling pathways. ERS treatment also recovered UVB-induced reduction of procollagen type I by activating the TGF-ß/Smad pathway. In addition, ERS exhibited an excellent antioxidant activity, which could increase the expression of cytoprotective antioxidants such as HO-1 and NQ-O1, by facilitating Nrf2 nuclear transfer. These findings demonstrated that the photoprotective effects of RS were significantly improved by enzyme-modified biotransformation.

Eucalyptus globulus extract protects against UVB-induced photoaging by enhancing collagen synthesis via regulation of TGF-ß/Smad signals and attenuation of AP-1.

UV irradiation triggers the overproduction of matrix metalloproteinases and collagen degradation, which in turn causes increased pigmentation, dryness, and deep wrinkling of the skin. These chronic symptoms are collectively referred to as photoaging. Eucalyptus globulus is an evergreen tree that is widely used in cosmetics because of its antimicrobial activity. In this study, we investigated the protective effect of 50% ethanol extracts of Eucalyptus globulus on UV-induced photoaging in vitro and in vivo. Normal human dermal fibroblasts were treated with Eucalyptus globulus at concentrations ranging from 1 to 100 µg/mL after UVB or non-UVB irradiation. We found that Eucalyptus globulus suppressed the expression of MMPs and IL-6, but increased the expression of TGF-ß1 and procollagen type 1. In addition, Eucalyptus globulus inhibited activation of the AP-1 transcription factor, an inducer of MMPs. Eucalyptus globulus was also found to regulate TGF-ß/Smad signaling by reversing the activity of negative Smad regulators. Lastly, in vivo studies showed that topical application of Eucalyptus globulus on UVB-irradiated hairless mice reduced wrinkle formation and dryness by down-regulating MMP-1 and up-regulating expression of elastin, TGF-ß1, and procollagen type 1. Taken together, these data suggest that Eucalyptus globulus may be a useful agent in cosmetic products.

Dietary Foeniculum vulgare Mill extract attenuated UVB irradiation-induced skin photoaging by activating of Nrf2 and inhibiting MAPK pathways.

BACKGROUND: Foeniculum vulgare Mill (FV) has long been prescribed in traditional medicine due to its antioxidant anti-inflammatory properties. However, little research has been done on the use of FV to alleviate changes in UVB-induced photoaging PURPOSE: This study was to investigate the photoprotective effects and mechanism of FV in vitro and in vivo. METHODS: The anti-photoaging effect of FV was assessed in normal human dermal fibroblasts (NHDFs) in vitro. The secretion of reactive oxygen species (ROS), lactate dehydrogenase (LDH), GSH, matrix metalloproteinases (MMPs), procollagen type I, IL-6 and transforming growth factor-ß1 (TGF-ß1) were measured by kits. Additionally, the level of nuclear factor (erythroid-derived 2)-like 2 (Nrf2), p-ERK and p38 were evaluated by western blotting. In vivo, H&E and Masson's trichrome staining were employed. The expression of MMP-1, procollagen type I, TGF-ß1 and elastin were measured by western blot. RESULTS: FV significantly increased the production of collagen, elastin and TGF-ß1 levels, while blocked matrix metalloproteinases (MMPs) production in UVB irradiation induced hairless mice, which were consistent with the result in NHDFs. Furthermore, FV dose-dependently decreased the production of ROS and LDH by promoting the nuclear amount of Nrf2 and enhancing the expression of cytoprotective antioxidants such as GSH. FV also significantly quenched UVB-induced phosphorylation of ERK and p38 in NHDFs. CONCLUSION: Our results indicate that FV is a potential botanical agent for the treatment of skin damage induced by UV irradiation.

Phytochemical constituents from the florets of tiger grass Thysanolaena latifolia from Nepal.

Phytochemical investigation on the florets of Thysanolaena latifolia leads to the isolation of a new compound 6″-O-acetylorientin-2″-O-α-L-rhamnopyranoside (1), named amrisoside and other 34 known compounds. The chemical structures of the compounds were determined from the interpretation of spectroscopic data including NMR, MS, and IR. This is the first report of phytochemical constituents from the monotypic genus Thysanolaena.

Identification of bitter components from Artemisia princeps Pamp.

Identification of bitter components from the aerial parts of Artemisia princeps Pamp. was performed to search for a method to eliminate the bitter taste from A. princeps products. The aerial parts of A. princeps were extracted in an aqueous EtOH solution, and the obtained extracts were partitioned into essential-oil, flavonoid-rich, n-BuOH, and aqueous fractions. Two purified bitter sesquiterpenoids were identified through repeated column chromatography of the bitterest fraction, the flavonoid-rich fraction, through an activity-guided fractionation method. The compounds were identified to be 1α,6α,8α-trihydroxy-5α,7ßH-guaia-3,9,11(13)-trien-12-oic acid and artecalin, respectively, based on the interpretation of NMR, MS, and IR spectroscopic data. Both compounds were 50 times bitterer than caffeine and had similar bitterness to quinine HCl. Neither eupatilin nor jaceosidin, the major active components of A. princeps, showed any bitterness.

Diterpenes from the roots of Oryza sativa L. and their inhibition activity on NO production in LPS-stimulated RAW264.7 macrophages.

Two new pimarane diterpenoids, momilactone D (3) and momilactone E (5), along with three known diterpenoids, momilactone A (1), sandaracopimaradien-3-one (2), and oryzalexin A (4) were isolated from Oryza sativa roots. The chemical structures of the compounds were determined by spectroscopic data analysis. The isolated diterpenoids were evaluated for their ability to inhibit NO production and iNOS mRNA and protein expression in LPS-stimulated RAW264.7 macrophages. Compound 4 showed strong inhibition activity on NO production, and compounds 1 and 4 decreased the expression of iNOS mRNA and protein levels.

Inhibition effect of phenyl compounds from the Oryza sativa roots on melanin production in murine B16-F10 melanoma cells.

Five phenyl compounds, vanillin (1), methyl trans-ferulate (2), trans-p-coumaric acid methyl ester (3), N-benzoyltryptamine (4), and N-(trans-cinnamoyl)tryptamine (5), were isolated from the roots of Oryza sativa L. and identified on the basis of spectroscopic data. Compounds 3 and 5 showed strong inhibition effect on melanin production in murine B16-F10 melanoma cells and tyrosinase activity. Also, the quantitative analysis of the compounds was carried out using LC/MS/MS experiment. Compounds 3 and 5 could be used as skin-whitening agents.

Lignan and flavonoids from the stems of Zea mays and their anti-inflammatory and neuroprotective activities.

The stems of Zea mays L., otherwise known as cornstalks, were extracted with 80 % aqueous MeOH, and the concentrated extract was successively partitioned with ethyl acetate (EtOAc), normal butanol, and water. From the EtOAc fraction, a new lignan along with three known flavonoids, tricin (1), salcolin A (2), and salcolin B (3), were isolated. The chemical structure of the lignan was determined to be tetrahydro-4,6-bis(4-hydroxy-3-methoxyphenyl)-1H,3H-furo[3,4-c]furan-1-one (4) through spectroscopic data analyses including NMR, MS, and IR. All compounds were isolated for the first time from this plant. The isolated compounds were evaluated for their inhibitory activity against NO production in Lipopolysaccharide-induced RAW 264.7 cells and their protective activity in glutamate-induced cell death in HT22 cells. The compounds 1, 2 and 4 showed anti-inflammatory effects with IC50 values of 2.63, 14.65, and 18.91 µM, respectively, as well as neuroprotective effects with EC50 values of 25.14, 47.44, and >80 µM, respectively.

Flavonoids from Machilus japonica stems and their inhibitory effects on LDL oxidation.

Stems of Machilus japonica were extracted with 80% aqueous methanol (MeOH) and the concentrated extract was successively extracted with ethyl acetate (EtOAc), normal butanol (n-BuOH), and water. Six flavonoids were isolated from the EtOAc fraction: (+)-taxifolin, afzelin, (-)-epicatechin, 5,3'-di-O-methyl-(-)-epicatechin, 5,7,3'-tri-O-methyl-(-)-epicatechin, and 5,7-di-O-methyl-3',4'-methylenedioxyflavan-3-ol. The chemical structures were identified using spectroscopic data including NMR, mass spectrometry and infrared spectroscopy. This is the first report of isolation of these six compounds from M. japonica. The compounds were evaluated for their diphenyl picryl hydrazinyl scavenging activity and inhibitory effects on low-density lipoprotein oxidation. Compounds 1 and 3-6 exhibited DPPH antioxidant activity equivalent with that of ascorbic acid, with half maximal inhibitory concentration (IC50) values of 0.16, 0.21, 0.17, 0.15 and 0.07 mM, respectively. The activity of compound 1 was similar to the positive control butylated hydroxytoluene, which had an IC50 value of 1.9 µM, while compounds 3 and 5 showed little activity. Compounds 1, 3, and 5 exhibited LDL antioxidant activity with IC50 values of 2.8, 7.1, and 4.6 µM, respectively.

Re-evaluation of physicochemical and NMR data of triol ginsenosides Re, Rf, Rg2, and 20-gluco-Rf from Panax ginseng roots.

Ginseng roots were extracted with aqueous methanol, and extracts were suspended in water and extracted successively with ethyl acetate and n-butanol. Column chromatography using the n-butanol fraction yielded four purified triol ginseng saponins: the ginsenosides Re, Rf, Rg2, and 20-gluco-Rf. The physicochemical, spectroscopic, and chromatographic characteristics of the ginsenosides were measured and compared with reports from the literature. For spectroscopic analysis, two-dimensional nuclear magnetic resonance (NMR) methods such as (1)H-(1)H correlation spectroscopy, nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond connectivity were employed to identify exact peak assignments. Some peak assignments for previously published (1)H- and (13)C-NMR spectra were found to be inaccurate. This study reports the complete NMR assignment of 20-gluco-Rf for the first time.

Procyanidins from the stem wood of Machilus japonica and their inhibitory effect on LDL oxidation.

The stem wood of Machilus japonica Siebold & Zucc were extracted with 80 % aqueous MeOH, and the concentrated extract was successively partitioned with ethyl acetate (EtOAc), normal butanol, and water. From the EtOAc fraction, five procyanidins, procyanidin A1 (1), procyanidin A2 (2), procyanidin B7 (3), cinnamtannin B1 (4), and aesculitannin B (5), were isolated. Their chemical structures were identified through spectroscopic data analyses including NMR, MS, and IR. This is the first time any of these compounds have been isolated from this plant. The compounds were evaluated for inhibition activity on LDL oxidation. All of these compounds and the positive control, BHT, showed a very high inhibition effect with IC50 values of 0.94, 2.1, 1.8, 1.1, 1.0, and 1.9 µM, respectively.

Flavonoids from the grains of C1/R-S transgenic rice, the transgenic Oryza sativa spp. japonica, and their radical scavenging activities.

The transgenic rice cultivar of Oryza sativa spp. japonica cv. Hwa-Young, C1/R-S transgenic rice (C1/R-S rice), is a flavonoid-rich cultivar of rice. The grains of C1/R-S rice were extracted with aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH, and H2O, successively. Repeated silica gel, octadecyl silica gel (ODS), and Sephadex LH-20 column chromatographies for the EtOAc and n-BuOH fractions afforded four new flavonoids (compounds 2, 3, 7, and 8) along with four known flavonoids: (+)-3'-O-methyltaxifolin (1), brassicin (4), isorhamnetin-4'-O-ß-D-glucosyranoside (5), and 3'-O-methyltaxifolin-5-O-ß-D-glucopyranoside (6). The new flavonoids were identified as 3'-O-methyltaxifolin-7-O-ß-D-glucopyranoside (2), 3'-O-methyltaxifolin-4'-O-ß-D-glucopyranoside (3), isorhamnetin-7-O-ß-D-cellobioside (brassicin-4″-O-ß-D-glucopyranoside) (7), and brassicin-4'-O-ß-D-glucosyranoside (8) from the result of spectroscopic data including nuclear magnetic resonance spectrometry (NMR), mass spectrometry (MS), and infrared spectroscopy (IR). Also, quantitative analysis of major flavonoids (compounds 2, 3, and 8) in C1/R-S rice, O. sativa spp. japonica cv. Hwa-Young (HY), and a hybrid of two cultivar (C1/R-S rice/HY) extracts was performed using HPLC experiment. The isolated flavonoids were evaluated for their radical-scavenging effect on DPPH and ABTS radicals.

Potential neuroprotective flavonoid-based inhibitors of CDK5/p25 from Rhus parviflora.

Rhus parviflora (Anacardiaceae) is an indigenous medicinal shrub found in South Asia with flavonoid rich edible fruit. This study examined flavonoid derivatives of R. parviflora fruit with CDK5/p25 inhibition activity. Evaluation by in vitro assay and docking simulations for CDK5/p25 revealed that the aurones, sulfuretin (1) and aureusidin (2), the aurone glycoside, aureusidin-6-O-ß-D-glucopyranoside (3) and hovetrichoside C (4), the flavonoid glycoside, quercetin-3-O-ß-D-galactopyranoside (5), and the biflavonoid, cupressuflavone (6), had the potential to inhibit CDK5/p25, which could be useful in the treatment of neurodegenerative disorders such as Alzheimer's disease. Compound2 showed the significant in vitro inhibition capacity (IC50 value of 4.81 µM) as well as binding affinity with docking energy of -8.73 (kcal/mol) for active sites CYS83 and GLN130 of CDK5/p25 enzyme in comparison to reference compound R-roscovitine.

Phenolic components from Rhus parviflora fruits and their inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW 264.7 macrophages.

Nine phenolic compounds, phloracetophenone-4-O-ß-D-glucopyranoside (1), p-hydroxybenzoic acid-4-O-ß-D-glucopyranoside (2), leonuriside A (3), 3-methoxy-4-hydroxyphenol-1-O-ß-D-glucopyranoside (4), cis-p-coumaric acid-4-O-ß-D-glucopyranoside (5), trans-p-coumaric acid-4-O-ß-D-glucopyranoside (6), trans-p-coumaric acid-9-O-ß-D-glucopyranoside (7), (-)-shikimic acid (8) and (-)-methyl shikimate (9), were isolated for the first time from the fruits of Rhus parviflora. Compounds 1, 3-6 and 8 inhibited lipopolysaccharide-stimulated nitric oxide (NO) production and inducible NO synthase expression in RAW 264.7 macrophages with IC50 values of 9.24 ± 1.20, 21.37 ± 2.02, 23.07 ± 1.58, 9.86 ± 0.98, 19.05 ± 1.66 and 11.3 ± 1.54 µM, respectively. The results indicated possible use of compounds for the treatment of inflammatory diseases.

Triterpenoids from Fragaria ananassa calyx and their inhibitory effects on melanogenesis in B16-F10 mouse melanoma cells.

Column chromatographic technology was applied to isolate six purified ursane triterpenoids from the calyx of Fragaria ananassa and they were identified on the basis of spectroscopic methods to be ursolic acid (1), pomolic acid (2), 2-oxo-pomolic acid (3), 3-O-acetyl pomolic acid (4), fupenzic acid (5) and euscaphic acid (6). This is the first study in which these compounds have been isolated from the calyx of F. ananassa. Compared to a well-known inhibitor, α-arbutin, compounds 2-6 showed a significant decrease in intracellular melanin content in B16-F10 cells, and in culture media melanin.

Carbohydrate derivatives from the roots of Brassica rapa ssp. campestris and their effects on ROS production and glutamate-induced cell death in HT-22 cells.

Phytochemical investigation of the roots of Brassica rapa ssp. campestris led to the isolation of three new carbohydrate derivatives, namely sucrose 3,3',4'-triisovalerate (2), sucrose 6,3',4'-triisovalerate (3), and ethanone-1-C-ß-d-glucopyranoside (3,7-anhydro-1-deoxy-d-glycero-d-gulo-2-octulose, 6), along with four known carbohydrate derivatives, 2,6,3',4'-tetraisovalerate (1), ethyl ß-d-glucopyranoside (4), n-butyl ß-d-fructofuranoside (5), and n-pentyl ß-d-fructofuranoside (7), which were initially isolated from plants of the Brassica genus. Structures of the isolated compounds were established by spectroscopic analyses, including UV, IR, MS, and NMR. All of the isolated carbohydrate derivatives were evaluated to determine their effect on ROS production and glutamate-induced cell death in HT-22 cells. Compound 6 showed the most significant ROS reduction and a protective effect with IC50 values of 69.4 ± 3.8 µM and 4.96 ± 0.32 µM, respectively, which were equivalent to those of the positive control, Trolox.

A new phenanthrene derivative and two diarylheptanoids from the roots of Brassica rapa ssp. campestris inhibit the growth of cancer cell lines and LDL-oxidation.

Brassica rapa ssp. campestris (Brassicaceae) is a conical, deep purple, edible root vegetable commonly known as a turnip. We initiated phytochemical and pharmacological studies to search for biological active compounds from the roots of B. rapa ssp. campestris. We isolated a novel phenanthrene derivative, 6-methoxy-1-[10-methoxy-7-(3-methylbut-2-enyl)phenanthren-3-yl]undecane-2,4-dione, named brassicaphenanthrene A (3) along with two known diarylheptanoid compounds, 6-paradol (1) and trans-6-shogaol (2), through the repeated silica gel (SiO2), octadecyl silica gel, and Sephadex LH-20 column chromatography. The chemical structures of the compounds were determined by spectroscopic data analyses including nuclear magnetic resonance, mass spectrometry, ultraviolet spectroscopy, and infra-red spectroscopy. All compounds exhibited high inhibitory activity against the growth of human cancer lines, HCT-116, MCF-7, and HeLa, with IC50 values ranging from 15.0 to 35.0 µM and against LDL-oxidation with IC50 values ranging from 2.9 to 7.1 µM.

Artemisia leaf extract induces apoptosis in human endometriotic cells through regulation of the p38 and NFκB pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia leaves have long been used for the treatment of gynecological disorders, including infertility and dysmenorrhea, which can be commonly caused by endometriosis. In the present study, we investigated the effect of Artemisia princeps extract (APE) on the cell growth and apoptosis of human endometriotic cells. MATERIALS AND METHODS: MTT assays and FACS analysis using PI and Annexin staining were performed to study cell viability, cell cycle progression, and apoptosis. We also explored the mechanism of APE-induced effects by evaluating the activation of caspases, Akt, p38, and NFκB. The expressions of XIAP, Bcl-2, and Bcl-xL were measured by real-time RT-PCR and Western blot analyses. RESULTS: APE significantly inhibited the cell viability of 11Z and 12Z human endometriotic epithelial cells. Interestingly, endometriotic cells were more sensitive to APE treatment than immortalized endometrial cells (HES). Treatment with APE induced apoptosis of 11Z cells in a time-dependent manner, as shown by accumulation of sub G1 and apoptotic cell populations. In addition, treatment with APE stimulated the activation of caspase -3, -8, and -9 in a dose- and time-dependent manner. Furthermore, p38 was activated by APE treatment, and the p38 inhibitor SB203580 markedly inhibited APE-induced cell death in 11Z cells. Moreover, treatment with APE suppressed the activation of NFκB and the expressions of anti-apoptotic factors such as XIAP, Bcl-2, and Bcl-xL. CONCLUSION: These results indicate that APE is a potential anti-endometriotic agent, acting to induce apoptosis of endometrial cells through the modulation of the p38 and NFκB pathways.