Effect of Tannin in Green Tea By-Product in Combination with Bio-Char Supplemented into Basal Beef Cattle Diet on Nutrient Digestibility, Methane Production and Animal Performance

The experiment was conducted by using 20 bulls of Lai Sind cattle (a cross breed between Red Sindhi and local Vietnam yellow cattle) from 15 - 16 months of age, weighed approximately 154 - 156 kg, housed individually with provision of adequate feed trough and free access to water to evaluate the effect of tannin sourced from green tea by-product and addition of bio-char on methane production and animal performance. The experimental diets were formulated by replacing elephant grass in basal diet by bio-char and green tea by-product with the replacement of 0.5 - 1 bio-char and 3% - 5% green tea by-products, respectively to make the content of tannin in the diet of 7.3 g and 12.5 g tannin per kg dry matter. Results from this study revealed that bio-char and green tea by-product included into the diet with different levels had significantly affected on the feed intake, as the increasing level of tannin in the diet tended to increase the feed intake. The inclusion of green tea by-product in the diet which resulted in the content of tannin in the diet of 12.5 g/kg DM had significantly reduced methane emission without altering animal performance, thus the inclusion of 0.5% bio-char and 5% of green tea by-product can be applied in order to mitigate methane emission, thus contributed to environment protection.

Effect of a Commercial Extract of Green Tea and a Pure Catechin on Two Veillonella strains

The catechin Epigallocatechin-3-O-Gallate (EGCG) which is found in of Green Tea extracts (GTE), displays a variety of microbicidal properties. It is largely believed that EGCG inhibits the growth of cariogenic and periodontopathic bacteria. Objective: In this paper we compared the inhibitory activity of EGCG and a commercial GTE on the growth of Veillonella parvula. Chlorhexidine was used as positive control. Methodology: V. parvula ATCC 10790 and a clinical isolate obtained from a periodontal disease patient were cultured in the presence of EGCG or a commercial GTE, and the measurements of bacterial growth inhibition were compared to the values obtained with 0.12 and 0.2% chlorhexidine. Results: Chlorhexidine inhibited bacterial growth, however in contrast to a previous report, neither EGCG nor the GTE showed any effect on bacterial growth. Conclusions: The data show and confirm that chlorhexidine is a growth inhibitor of V. parvula while EGCG and GTE do not display such effect.

Synergistic effects of three traditional herbs green tea,mulberry leaf and corn silk on glucose uptake level of L6 myoblasts and the hypoglycemic mechanism

Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.

Hypoglycemic Effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi Green Tea and Their Compatibility on Mice

[Objectives]To explore the hypoglycemic effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi green tea and their compatibility on mice,so as to provide more options for the adjuvant treatment of diabetes and improve the comprehensive utilization value of Guixi green tea.[Methods]After being fed adaptively for 7 d,60 SPF-grade male mice were intraperitoneally injected with alloxouracil(180 mg/kg)to induce hyperglycemia model,from which 35 mice with a successful model were selected.They were randomly divided into 5 groups(Danshen group,green tea group,compatible group,positive control group and blank control group),with 7 animals in each group.The mice in all groups were administered according to the dose of 200 mg/kg once a day for 15 d.After the last administration,their body weight was measured on the 15 th day after fasting for 12 h,and their fasting blood glucose was measured by tail clipping.[Results]After modeling,the mice in each group showed typical hyperglycemia symptoms,namely obvious polydipsia,polyphagia,polyuria and weight loss.Compared with the blank control group,Danshen group and green tea group had obvious hypoglycemic effect(P<0.05),and compatible group had significant hypoglycemic effect(P<0.01).Compared with the blank control group,the weight loss of diabetic mice could also be inhibited in green tea group and compatible group(P<0.05).[Conclusions]Danshen,Guixi green tea and their compatibility had the effect of lowering blood sugar,among which the compatible group had the most obvious effect.

Assessing the effectiveness of nanoscale zero-valent iron particles produced by green tea for Cr(VI)-contaminated groundwater remediation

Nanoscale zero-valent iron particles(NZVI) produced by using green tea(GT) extract as a reductant can remove Cr(Ⅵ) from water effectively,which can be utilized in groundwater remediation.In order to define the reaction mechanism and removal effect in the aquifer,in this study,GT-NZVI particles were prepared and measured by some characterization methods to define their surface performance,and then batch and one-dimensional experiments were carried out to reveal the reaction properties of GT-NZVI and Cr(Ⅵ) in groundwater.The results showed that the prepared GT-NZVI particles were regular spherical with a diameter of 10-20 nm,which could disperse in water stably.The main component of GT-NZVI wasα-Fe with superficial polyphenols as a stabilizer.GT-NZVI suspension had good ability to reduce the Cr(Ⅵ) to Cr(Ⅲ) in water.When the concentration of GT-NZVI was 1 g/L,the removal efficiency of Cr(Ⅵ)with an initial concentration of 100 mg/L reached 92.8% in 1 h reaction.In column tests,GT-NZVI passed through the natural sand column successfully with an average outflow percentage of 71.2%.The simulated in-situ reaction zone(IRZ) with GT-NZVI was used to remediate Cr(Ⅵ) contaminated groundwater.The oufflow concentration of Cr(Ⅵ) kept in 0.14-0.32 mg/L corresponding to the outflow rate below 0.32%within 15 days,and the removal efficiency of Cr(Ⅵ) by IRZ with GT-NZVI decreased with the increase of aquifer medium particle size,groundwater flow rate and ionic strength.Most of Cr(Ⅲ) as reduzate was adsorbed or immobilized on the surface or in the lattice of GT-NZVI,which indicated effective immobilization for chromium.

Green tea polyphenols alleviate di-(2-ethylhexyl)phthalate-induced liver injury in mice

BACKGROUND Di(2-ethylhexyl)phthalate(DEHP)is a common plasticizer known to cause liver injury.Green tea is reported to exert therapeutic effects on heavy metal exposureinduced organ damage.However,limited studies have examined the therapeutic effects of green tea polyphenols(GTPs)on DEHP-induced liver damage.AIM To evaluate the molecular mechanism underlying the therapeutic effects of GTPs on DEHP-induced liver damage.METHODS C57BL/6J mice were divided into the following five groups:Control,model[DEHP(1500 mg/kg bodyweight)],treatment[DEHP(1500 mg/kg bodyweight)+GTP(70 mg/kg bodyweight),oil,and GTP(70 mg/kg bodyweight)]groups.After 8 wk,the liver function,blood lipid profile,and liver histopathology were examined.Differentially expressed micro RNAs(miRNAs)and mRNAs in the liver tissues were examined using high-throughput sequencing.Additionally,functional enrichment analysis and immune infiltration prediction were performed.The miRNA-mRNA regulatory axis was elucidated using the starBase database.Protein expression was evaluated using immunohistochemistry.RESULTS GTPs alleviated DHEP-induced liver dysfunction,blood lipid dysregulation,fatty liver disease,liver fibrosis,and mitochondrial and endoplasmic reticulum lesions in mice.The infiltration of macrophages,mast cells,and natural killer cells varied between the model and treatment groups.mmu-miR-141-3p(a differentially expressed miRNA),Zcchc24(a differentially expressed mRNA),and Zcchc24(a differentially expressed protein)constituted the miRNA-mRNA-protein regulatory axis involved in mediating the therapeutic effects of GTPs on DEHP-induced liver damage in mice.CONCLUSION This study demonstrated that GTPs mitigate DEHP-induced liver dysfunction,blood lipid dysregulation,fatty liver disease,and partial liver fibrosis,and regulate immune cell infiltration.Additionally,an important miRNAmRNA-protein molecular regulatory axis involved in mediating the therapeutic effects of GTPs on DEHP-induced liver damage was elucidated.

Green Synthesis of Iron(Ⅱ,Ⅲ)-polyphenol Nanoparticles and Their Adsorption of Malachite Green

Three kinds of iron nanoparticles(FeNPs)were prepared via green route based on pomegranate(PG),green tea(GT),and mulberry(ML)extracts under ambient conditions.The obtained materials were characterized by scanning electron microscopy(SEM),transmission electronic microscopy(TEM),X-ray energy-dispersive spectrometer(EDS),X-ray diffraction(XRD),fourier transform infrared spectroscopy(FTIR),and X-ray photoelectron spectroscopy(XPS)techniques.The experimental results show that FeNPs were in the form of amorphous iron(Ⅱ,Ⅲ)-polyphenol complex with different dispersity and morphologies.GT-Fe has the smallest size range of 25-35 nm,PG-Fe has a moderate size-distribution of 30-40 nm,while ML-Fe formed a tuberous net-type with a sheeting structure.PG-Fe displays the highest removal efficiency of 90.2%in 20 min towards cationic dye of malachite green(16.6%by ML-Fe and 69.3%by GT-Fe),which is attributed to its highest polyphenol content,lowest zeta potential,as well as the most Fe^(2+)on the surface of FeNPs.The removal mechanism was mainly induced by electrostatic adsorption based on pH and zeta potential tests.

Study on Processing Technology of Health Beverage Made from Artemisia Selengensis Turoz and Green Tea

This study took Artemisia selengensis Turoz and green tea as primary materials and sugar, lemon acid potassium as supplementary materials to make compound health beverage.The main influence factors to the quality of the compound beverage were investigated,such as addition amounts ofArtemisia selengensis juice, green tea juice,sugar and lemon acid.By orthogonal test ,the best formula of the compound health beverage is Artemisia selengensis juice 10 ml, green tea juice 200 ml, sugar 6 g and lemon acid 0.06 g. The optimum stabilizer of the beverage is CMC-Na with 0.03% amount.

DNA Extraction and Molecular Identification of Green Tea

[Objective] The aim of this study was to provide reference for the quality identification of green tea.[Method] Green Tea was used as materials,and its total DNA was extracted through improved CTAB method.And the obtained DNA was used to carry out identification on 10 varieties of green tea through ISSR molecular markers.[Result] The high quality DNA from green tea could be obtained with new method,the DNA yield ranged from 101-498 μg/g tea sample for various green tea samples,and the average yield was 249 μg/g tea sample.The ISSR detection result showed that ISSR markers could effectively differentiate different varieties of green tea.[Conclusion] The result had provided reference for the further study on molecular identification of green tea.

Exploiting push-pull strategy to combat the tea green leafhopper based on volatiles of Lavandula angustifolia and Flemingia macrophylla

Thirteen volatile compounds were identified from Flemingia macrophylla plants. Eight major components significantly attracted the tea green leafhoppers, Empoasca flavescens F. Based on their relative abundances, following synthetic blends were made for field experiments: 1) eight-component-attractant blend included Z-3-hexen-1-ol, Z-3-hexenyl acetate, Z-ocimene, Me SA, Z-3-hexenyl butyrate, dodecane, hexadecane and nonanal at 10, 10, 1, 11, 2, 6, 2 and 4 mg mL^-1 in n-hexane, respectively;2) four-component-attractant blend #1 contained hexadecane, Z-3-hexenyl acetate, Z-3-hexen-1-ol and nonanal at 2, 10, 10 and 4 mg mL^-1 in n-hexane, respectively;3) four-component-attractant blend #2 contained hexadecane, Z-3-hexenyl acetate, Z-3-hexen-1-ol and Me SA at 2, 10, 10 and 11 mg mL^-1 in n-hexane, respectively. Thymol and 1-methoxy-4-methyl-2-(1-methylethyl)-benzene, identified from Lavandula angustifolia aeration samples, significantly repelled the leafhopper as strong repellents when tested alone or in combination at 10 mg mL^-1. For field bioassays, each attractant lure was attached to a bud green sticky board hung from a bamboo stick at above tea plant level for catching the leafhoppers, whereas the repellent dispenser was tied to a tea branch inside tea clump for pushing the leafhoppers away from tea clumps. The results showed that the eight-component-attractant blend caught similar numbers of the leafhopper as did the four-component-attractant blend #1 at about 53–79 leafhoppers/trap/day, which were significantly higher than those on the hexane-control bud green sticky boards. Average leafhopper catches from un-baited sticky boards were about 51–73 leafhoppers/trap/day when pushed by the repellents placed inside tea plants, with the two-component-repellent blend being more effective than their single components. When the two-component-repellent blend was further tested with the three attractant blends in a push-pull fashion, average trap catches ranged from 62 to 92 leafhoppers/trap/day. Control efficacy on the leafhoppers within the push-pull zones increased progressively from day 1(43%) to day 5(73%). This push-pull approach might have a great potential as a green control strategy for combating the tea green leafhoppers.

Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa

Objective:To evaluate antibacterial activity of the Indonesian water soluble green tea extract,Camellia sinensis,against clinical isolates of methicillin-resistantStaphylococcus aureus (S.aureus)(MRSA)and multi-drug resistant Pseudomonas aeruginosa(MDR-P.aeruginosa).Methods:Antimicrobial activity of green tea extract was determined by the disc diffusion method and the minimum inhibitory concentration(MIC)was determined by the twofold serial broth dilutions method.The tested bacteria using in this study were the standard strains and multi-drug resistant clinical isolates of S.aureus and P.aeruginosa,obtained from Laboratory of Clinical Microbiology,Faculty of Medicine,University of Indonesia.Results:The results showed that the inhibition zone diameter of green tea extracts forS.aureus ATCC 25923 and MRSA were(18.970依0.287)mm,and(19.130依0.250)mm respectively.While the inhibition zone diameter forP.aeruginosa ATCC 27853 and MDR-P.aeruginosawere(17.550依0.393)mm and(17.670依0.398)mm respectively.The MIC of green tea extracts againstS.aureus ATCC 25923 and MRSA were 400μg/mL and 400μg/mL,respectively,whereas the MIC for P.aeruginosa ATCC 27853 and MDR-P.aeruginosawere 800μg/mL,and 800μg/mL,respectively.Conclusions:Camellia sinensisleaves extract could be useful in combating emerging drug-resistance caused by MRSA andP.aeruginosa.

Green tea and the risk of gastric cancer: Epidemiological evidence

Gastric cancer (GC) is one of the leading causes of cancer death in the world. Numerous efforts are being made to find chemoprotective agents able to reduce its risk. Amongst these, green tea has been reported to have a protective effect against stomach cancer. This article aims to critically evaluate all epidemiological studies reporting an association between green tea consumption and GC risk. MEDLINE, EBSCOHOST and Google Scholar were used to search for clinical trials of green tea and its correlation to stomach cancer. Studies include cohort and case-control studies. Outcome of interests are inverse association, no association, and positive association. Seventeen epidemiologic studies were reviewed. Eleven studies were conducted in Japan, five in China, and one with Japanese descendent in Hawaii. Ten case-control studies and seven cohortstudies were included. The relative risks or odds ratio of GC for the highest level of green tea consumption was compared. Seven studies suggested no association, eight an inverse association, and one a positive association. One study had shown a significantly lowered GC risk when tea was served warm to cold. Another study also showed a significantly risk with lukewarm tea. All studies that analyzed men and women separately have suggested a reduced risk in women than in men, albeit no significant difference. This review demonstrates that there is insufficient information to support green tea consumption reduces the risk of GC. More studies on the subject matter are warranted.

Effects of Chemical Components on the Amount of Green Tea Cream

The relationship between the amount of tea cream and the chemical components contents in the original green tea infusion was investigated. The results showed that the amounts of tea cream in the green tea infusions obtained from different cultivars and different parts of new shoots were varied. There were many chemical components participating in the formation of green tea cream. However, there were only the contents of caffeine （Y=0.85, P〈0.01） and polyphenols （Y=0.65, P〈0.05） in the original green tea infusion highly correlated with the amount of green tea cream. Stepwise regression analysis of overall chemical components indicated that the contents of caffeine and gallated catechins in the original green tea infusion had a significant effect （P〈0.01） on green tea cream levels. Cream （g L-1）=-172.071＋ 0.129×Ccaffeine＋0.024×Ggallated catechins （R2=0.936）. The amount of green tea cream can be predicted by the contents of gallated catechins and caffeine in the original tea infusion. Principal component analysis also indicated that catechins, minerals, and polysaccharides were the important chemical components in the formation of green tea cream.

Green tea compounds in breast cancer prevention and treatment

Breast cancer is the most common cancer among women. In recent years, many in vitro and in vivo studies indicate that green tea possesses anti-cancer effects. The epidemiological studies, however, have produced inconclusive results in humans. Likewise, results from animal models about the preventive or therapeutic effects of green tea components are inconclusive. The mechanisms by which green tea intake may influence the risk of breast cancer in humans remain elusive. Here, we review recent studies of green tea polyphenols and their applications in the prevention and treatment of breast cancer. Furthermore, we discuss the effect of green tea components on breast cancer by reviewing epidemiological studies, animal model studies and clinical trials. At last, we discuss the mechanisms by which green tea components suppress the develop-ment and recurrence of breast cancer. A better understanding of the mechanisms will improve the utilization of green tea in breast cancer prevention and therapy and pave the way to novel prevention and treatment strategies for breast cancer.

Green tea polyphenols ameliorate non-alcoholic fatty liver disease through upregulating AMPK activation in high fat fed Zucker fatty rats

AIM To investigate protective effects and molecular mechanisms of green tea polyphenols(GTP) on nonalcoholic fatty liver disease(NAFLD) in Zucker fatty(ZF) rats.METHODS Male ZF rats were fed a high-fat diet(HFD) for 2 wk then treated with GTP(200 mg/kg) or saline(5 m L/kg) for 8 wk, with Zucker lean rat as their control. At the end of experiment, serum and liver tissue were collected for measurement of metabolic parameters, alanine aminotransferase(ALT) and aspartate aminotransferase(AST), inflammatory cytokines and hepatic triglyceride and liver histology. Immunoblotting was used to detect phosphorylation of AMP-activated protein kinase(AMPK) acetyl-Co A carboxylase(ACC), and sterol regulatory element-binding protein 1c(SREBP1c). RESULTS Genetically obese ZF rats on a HFD presented with metabolic features of hepatic pathological changes comparable to human with NAFLD. GTP intervention decreased weight gain(10.1%, P = 0.052) and significantly lowered visceral fat(31.0%, P < 0.01). Compared with ZF-controls, GTP treatment significantly reduced fasting serum insulin, glucose and lipids levels. Reduction in serum ALT and AST levels(both P < 0.01) were observed in GTP-treated ZF rats. GTP treatment also attenuated the elevated TNFα and IL-6 in the circulation. The increased hepatic TG accumulation and cytoplasmic lipid droplet were attenuated by GTP treatment, associated with significantly increased expression of AMPK-Thr172(P < 0.05) and phosphorylated ACC and SREBP1c(both P < 0.05), indicating diminished hepatic lipogenesis and triglycerides out flux from liver in GTP treated rats. CONCLUSION The protective effects of GTP against HFD-induced NAFLD in genetically obese ZF rats are positively correlated to reduction in hepatic lipogenesis through upregulating the AMPK pathway.

Green tea extract:A potential cause of acute liver failure

The use of herbal products has increased significantly in recent years.Because these products are not subject to regulation by the Food and Drug Administration and are often used without supervision by a healthcare provider,the indication for and consumption of these supplements is quite variable.Moreover,their use is generally regarded as safe and natural by the lay-public.Unfortunately,there has been an increase in the number of reported adverse events occurring with the use of herbal products.We present a case of acute impending liver failure in an adolescent male using a weightloss product containing green tea extract.Our case adds to the growing concern surrounding the ingestion of green tea extract and serves to heighten healthcare provider awareness of a potential green tea extract hepatotoxicity.Despite the generally touted benefits of green tea as a whole,clinical concern regarding its use is emerging and has been linked to its concentration in multiple herbal supplements.Interestingly,the suspected harmful compounds are those previously proposed to be advantageous for weight-loss,cancer remedy,and anti-inflammatory purposes.Yet,we emphasize the need to be aware of not just green tea extract,but the importance of monitoring patient use of all dietary supplements and herbal products.

Comparative study on Leaf Tissue Morphologic Changes of Chang-Sheng-Chuan Green Brick Tea Before ad After Fermentation

[Objective] The aim was to understand the change of fermentation pro- cess of Chang-sheng-chuan green brick tea and analyze the changes in the con- tents of structure components of tea leaves before and after fermentation. [Method] Ten tea leaves before and after fermentation were chosen. The tea leaves sections were made using paraffin microtomy, PAS staining, and observed under light mi- croscopy. Through image analysis for the tea leaves, we compared the changes on the areas of the cellulose and colloid in cell before and after pile-fermentation. At the same time, the tea leaves were observed under transmission electron micro- scope. [Result] The results showed that the pile-fermentation process caused the tea leaves tissue to become loose, and expanded the cell space. Besides, the cell wall of tea leaves was obviously thinning, and even breaking which composed of cellulose. Image analysis showed that the leaf fiber composition reduced significant- ly, with a corresponding increase of yellow colloid in cell. Under observation of transmission electron microscope, there were large amounts of fungi located in the space between the colloid and the cell wall of the tea leaves after pile-fermentation. [Conclusion] The fermentation of Green brick tea is the process of microbial bio- transformation of tea, which is dominated by microbe and consumed the cellulose components of tea leaves.

Green tea polyphenols inhibit testosterone production in rat Leydig cells

This study investigated the acute effects of green tea extract （GTE） and its polyphenol constituents, （-）-epigallocatechin-3-gallate （EGCG） and （-）-epicatechin （EC）, on basal and stimulated testosterone production by rat Leydig cells in vitro. Leydig cells purified in a Percoll gradient were incubated for 3 h with GTE, EGCG or EC and the testosterone precursor androstenedione, in the presence or absence of either protein kinase A （PKA） or protein kinase C （PKC） activators. The reversibility of the effect was studied by pretreating cells for 15 min with GTE or EGCG, allowing them to recover for 1 h and challenging them for 2 h with human chorionic gonadotropin （hCG）, luteinizing hormone releasing hormone （LHRH）, 22（R）-hydroxycholesterol or androstenedione. GTE and EGCG, but not EC, inhibited both basal and kinase-stimulated testosterone production. Under the pretreatment conditions, the inhibitory effect of the higher concentration of GTE/EGCG on hCG/LHRH-stimulated or 22（R）- hydroxycholesterol-induced testosterone production was maintained, whereas androstenedione-supported testosterone production returned to control levels. At the lower concentration of GTE/EGCG, the inhibitory effect of these polyphenols on 22（R）-hydroxycholesterol-supported testosterone production was reversed. The inhibitory effects of GTE may be explained by the action of its principal component, EGCG, and the presence of a gallate group in its structure seems important for its high efficacy in inhibiting testosterone production. The mechanisms underlying the effects of GTE and EGCG involve the inhibition of the PKA/PKC signalling pathways, as well as the inhibition of P450 side-chain cleavage enzyme and 17β-hydroxysteroid dehydrogenase function.

Qualitative analysis of catechins from green tea GMB-4 clone using HPLC and LC-MS/MS

Objective:To identify the bioactive compounds in catechins isolation and its components from green tea GMB-4 clone.Methods:Green tea GMB-4 clones were extracted with distilled water at 90C.Samples were eluted into the column with 10%ethanol.Subsequently,the column was eluted with95%ethanol and evaporated separately.Green tea extract was identified by thin layer chromatography.Catechins were separated by the stationary phase in column chromatography using polyamide with 10%ethanol eluent and 95%ethanol.The results of isolations were analyzed by high performance liquid chromatographic(HPLC)and LCMS/MS.Analysis of catechins by HPLC was done by external standard.Results:Fraction from 10%ethanol showed that four major peaks at retention time of1.663,2.367,2.950 and 4.890,indicated the presence of four catechins components including catechin,epicatechins,gallocatechin and epigallocatechin.Whereas,fraction from 95%ethanol showed two main peaks at retention time of 5.167 and 9.82,which indicated the presence of epigallocatechin gallate(EGCG)and epicatechin gallate(ECG).EGCG(m/z 459),epigallocatechin(m/z 307),ECG(m/z 443),and epicatechin(m/z 291)were isolated and separated successfully using HPLC and LC-MS/MS.Conclusions:The HPLC and LC-MS/MS methods were successfully tuned for the qualitative analysis of green tea extract with EGCG and ECG.Four major catechins were separated and identified by LC-MS/MS,such as EGCG,epigallocatechin,ECG and epicatechin.The result of HPLC analysis showed that EGCG and ECG were main components from catechins isolation of green tea GMB-4 clone.

Synergistic antioxidant interactions between green tea and Ocimum gratissimum

Objective: To evaluate the antioxidant interactions between aqueous infusions of green tea and Ocimum gratissimum at different ratios. Methods: Antioxidant activities of aqueous infusion of green tea and Ocimum gratissimum (leaves) alone or in combination at various proportions (3:1, 2:1, 1:1, 1:2, 1:3) were determined by DPPH, ABTS, NO and ex-vivo assays including lipid peroxidation and haemolysis. Total phenolic content and flavonoid content was calculated by Folin-Ciocalteu reagent and aluminum chloride colorimetry method, respectively. A correlation study was also conducted between the antioxidant activity and total phenolic/flavonoid content of various infusions. The interactions were analyzed by combination index applying CompuSyn software. Results: Green tea exhibited high radical scavenging ability as compared to Ocimum gratissimum infusion. Combination of green tea and Ocimum gratissimum exhibited moderate antagonism to strong synergistic interaction at various ratios. A strong correlation was found between total phenolic content/total flavonoid content and antioxidant activities of individual infusions (green tea and Ocimum gratissimum). For binary mixture at different ratios, a weak to strong correlation was observed between total phenolic content and antioxidant activity and almost no correlation between total flavonoid content and antioxidant potential. Conclusions: Overall, green tea and Ocimum gratissimum combination (1:1) displayed the highest antioxidant potential and maximum synergism.