Comparative studies on the hypolipidemic and growth suppressive effects of oolong, black, pu-erh, and green tea leaves in rats

The four major commercial teas, oolong, black, pu-erh, and green teas, have been manufactured in southeast Asia. In this study, we evaluated the growth suppressive and hypolipidemic effect of these four different tea leaves by oral feeding to male Sprague-Dawley rats for 30 weeks. The results showed that the suppression of body weights of tea leaves-fed groups were in the order: oolong tea > pu-erh tea > black tea > green tea. Pu-erh tea and oolong tea could lower the levels of triglyceride more significantly than that of green tea and black tea, but pu-erh tea and green tea were more efficient than oolong tea and black tea in lowering the level of total cholesterol. In lipoprotein, 4% pu-erh tea could increase the level of HDL-C and decrease the level of LDL-C, but other teas simply decrease the levels of both. The activity of antioxidant enzyme SOD is increased in all tea-fed groups as compared to the basal diet-fed group. Finally, relative weight ratios of liver to epididylmal adipose tissue were lower in feeding oolong tea and pu-erh tea groups. On the basis of these findings, it seemed that the fully fermented pu-erh and black tea leaves and partially fermented oolong tea leaves were more effective on their growth suppressive and hypolipidemic effects as compared to the nonfermented green tea leaves.     

Structure-activity relationships of tea compounds against human cancer cells

The content of the biologically active amino acid theanine in 15 commercial black, green, specialty, and herbal tea leaves was determined as the 2,4-dinitrophenyltheanine derivative (DNP-theanine) by a validated HPLC method. To define relative anticarcinogenic potencies of tea compounds and teas, nine green tea catechins, three black tea theaflavins, and theanine as well as aqueous and 80% ethanol/water extracts of the same tea leaves were evaluated for their ability to induce cell death in human cancer and normal cells using a tetrazolium microculture (MTT) assay. Compared to untreated controls, most catechins, theaflavins, theanine, and all tea extracts reduced the numbers of the following human cancer cell lines: breast (MCF-7), colon (HT-29), hepatoma (liver) (HepG2), and prostate (PC-3) as well as normal human liver cells (Chang). The growth of normal human lung (HEL299) cells was not inhibited. The destruction of cancer cells was also observed visually by reverse phase microscopy. Statistical analysis of the data showed that (a) the anticarcinogenic effects of tea compounds and of tea leaf extracts varied widely and were concentration dependent over the ranges from 50 to 400 microg/mL of tea compound and from 50 to 400 microg/g of tea solids; (b) the different cancer cells varied in their susceptibilities to destruction; (c) 80% ethanol/water extracts with higher levels of flavonoids determined by HPLC were in most cases more active than the corresponding water extracts; and (d) flavonoid levels of the teas did not directly correlate with anticarcinogenic activities. The findings extend related observations on the anticarcinogenic potential of tea ingredients and suggest that consumers may benefit more by drinking both green and black teas.     

Total phenol, catechin, and caffeine contents of teas commonly consumed in the United kingdom.

Levels of total phenol, catechins, and caffeine in teas commonly consumed in the United Kingdom have been determined using reversed phase high-performance liquid chromatography. Tea bags or tea leaves were purchased from local supermarkets and extracted in boiling water for 5 min. The resulting data showed considerable variability in both total phenols [80.5-134.9 mg/g of dry matter (DM) in black teas and 87-106.2 mg/g of DM in green teas] and catechins (5.6-47.5, 51.5-84.3, and 8.5-13.9 mg/g of DM in black, green, and fruit teas, respectively); this was most probably a result of differing agronomic conditions, leaf age, and storage during and after transport, as well as the degree of fermentation. Caffeine contents of black teas (22-28 mg/g of DM) were significantly higher than in less fermented green teas (11-20 mg/g of DM). The relative concentration of the five major tea catechins ranked EGCG > ECG > EC > EGC > C. The estimated U.K. dietary intakes of total tea catechins, calculated on the basis of an average tea consumption of three cups of tea (200 mL cup, 1% tea leaves w/v), were 61.5, 92.7, and 405.5 mg/day from fruit teas, black teas, and green teas, respectively. The coefficients of variation were 19.4, 88.6, and 17.3%, respectively, indicating the wide variation in these intakes. The calculated caffeine intake ranged between 92 and 146 mg/day. In addition, many individuals will consume much larger quantities of tea, of various strengths (as determined by the brewing conditions employed). This broad spread of U.K. daily intakes further emphasizes the need for additional research to relate intake and effect in various population groups.     

Profiles of purine and pyrimidine nucleotides in fresh and manufactured tea leaves

Profiles of nucleotide levels in two varieties of Japanese green teas (cv. Yabukita and Saemidori), a Chinese green tea (Longjing), and two Japanese black teas (cv. Benifuuki and Benihikari) were determined and compared with that of fresh tea leaves. The concentration of 5'-nucleotides in green tea was much higher than in black tea. Nucleoside diphosphates were present in larger amounts than nucleoside triphosphates in manufactured green and black teas, whereas the triphosphates predominated in fresh tea leaves. Low levels of 3'-nucleotides were found in green and black teas. Inosine 5'-monophosphate, which is utilized as a seasoning component, was found in all manufactured teas in concentrations ranging from 50 to 200 nmol/g of dry weight. The levels of both inosine 5'-monophosphate and guanosine 5'-monophosphate were high in Chinese Longjing green tea. The unique profiles of nucleotides in manufactured teas may be a consequence of the action of degradation enzymes, such as ribonuclease, apyrase, phosphatase, nucleotidase, and adenosine 5'-monophsphate deaminase during the commercial processing of the young leaves.     

New phenolic components and chromatographic profiles of green and fermented teas

A standardized profiling method based on liquid chromatography with diode array and electrospray ionization mass spectrometric detection (LC-DAD-ESI/MS) was applied to establish the phenolic profiles of 41 green teas and 25 fermented teas. More than 96 phenolic compounds were identified that allowed the teas to be organized into five groups. Epigallocatechin gallate (EGCG) was the major phenolic component of green tea made from mature leaves (group 2), while green tea made from the younger buds and leaves (group 1) contained lower flavonoid concentrations. Partially fermented teas (group 3) contained one-half the EGCG content of the green tea. Fully fermented black teas (group 4) had a trace of EGCG, but contained theaflavins. Highly overfermented black tea (group 5) contained only trace amounts of flavonol glycosides and theaflavins. Over 30 phenolics are new for tea, and this is the first phenolic profile to simultaneously detect C- and O-glycosylated flavonoids, catechins, proanthocyanidins, phenolic acid derivatives, and purine alkaloids.     

Determination of theanine, GABA, and other amino acids in green, oolong, black, and Pu-erh teas with dabsylation and high-performance liquid chromatography

Dabsyl chloride (dimethylaminoazobenzene sulfonyl chloride), a useful chromophoric labeling reagent for amino acids and amines, was developed in this laboratory in 1975. Although several methods have been developed to determine various types of amino acids, a quick and easy method of determining theanine, GABA, and other amino acids has not been developed in one HPLC system. In this paper are analyzed the free amino acid contents of theanine and GABA in different teas (green tea, black tea, oolong tea, Pu-erh tea, and GABA tea) with a dabsylation and reverse phase high-performance liquid chromatography (HPLC) system coupled with a detector at 425 nm absorbance. Two reverse phase columns, Hypersil GOLD and Zorbax ODS, were used and gave different resolutions of dabsyl amino acids in the gradient elution program. The data suggest that the tea source or the steps of tea-making may contribute to the theanine contents variations. High theanine contents of high-mountain tea were observed in both green tea and oolong tea. Furthermore, the raw (natural fermented) Pu-erh tea contained more theanine than ripe (wet fermented) Pu-erh tea, and the GABA contents in normal teas were generally lower than that in GABA tea.     

Antioxidative activities of volatile extracts from green tea, oolong tea, and black tea

Antioxidative activities of volatile extracts from six teas (one green tea, one oolong tea, one roasted green tea, and three black teas) were investigated using an aldehyde/carboxylic acid assay and a conjugated diene assay. The samples were tested at levels of 20, 50, 100, and 200 micrograms/mL of dichloromethane. The results obtained from the two assays were consistent. All extracts except roasted green tea exhibited dose-dependent inhibitory activity in the aldehyde/carboxylic acid assay. A volatile extract from green tea exhibited the most potent activity in both assays among the six extracts. It inhibited hexanal oxidation by almost 100% over 40 days at the level of 200 micrograms/mL. The extract from oolong tea inhibited hexanal oxidation by 50% in 15 days. In the case of the extract from roasted green tea, the lowest antioxidative activity was obtained at the level of 200 micrograms/mL, suggesting that the extract from roasted green tea contained some pro-oxidants. The extracts from the three black teas showed slight anti- or proactivities in both assays. The major volatile constituents of green tea and roasted green tea extracts, which exhibited significant antioxidative activities, were analyzed using gas chromatography-mass spectrometry. The major volatile chemicals with possible antioxidative activity identified were alkyl compounds with double bond(s), such as 3,7-dimethyl-1,6-octadien-3-ol (8.04 mg/kg), in the extract from green tea and heterocyclic compounds, such as furfural (7.67 mg/kg), in the extract from roasted green tea. Benzyl alcohol, which was proved to be an antioxidant, was identified both in a green tea extract (4.67 mg/kg) and in a roasted tea extract (1.35 mg/kg).     

Screening for antioxidant activity in edible plant products: comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin-Ciocalteu assay

Oxidation of low-density lipoprotein (LDL) has been implicated in atherogenesis. Antioxidants that prevent LDL from oxidizing may reduce atherosclerosis. This study investigated LDL antioxidant activity in edible plant products for development of dietary supplementation to prevent atherosclerosis. Fifty-two kinds of edible plants were extracted using 70% aqueous ethanol solution, and the antioxidant activity of the extracts, which inhibit human LDL oxidation induced by copper ion, was determined on the basis of the oxidation lag time and represented as epigallocatechin 3-gallate equivalent. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenolic content were also measured for comparisons with antioxidant activity in LDL. Plant products showing the greatest activity in LDL oxidation assay were akamegashiwa (Mallotus japonicus) leaf, Japanese privet (Ligustrum japonicum) leaf, green tea [Camellia sinensis (L.) O. Kuntze], and astringent persimmon (Diospyros kaki). The present study revealed high levels of LDL antioxidant activity in plant products for which such activity levels are underestimated in the DPPH radical scavenging assay and Folin-Ciocalteu assay.     

Comparison of the antioxidant activity of commonly consumed polyphenolic beverages (coffee, cocoa, and tea) prepared per cup serving.

In this study, the in vitro low-density lipoprotein oxidation model was used to assess the relative antioxidant activity of the polyphenolic beverages tea, coffee, and cocoa on a cup-serving basis. The beverages were prepared as 0.7-2.5% soluble coffee and 1.5-3.5% cocoa; teas (green, black, or herbal) were prepared as one tea bag infused over 5 min in 220 mL of hot water. Under these standard cup serving conditions, the antioxidant activity as determined by the lag time was in the range of 292-948 min for coffee, 217-444 min for cocoa, 186-338 min for green tea, 67-277 min for black tea, and 6-78 min for herbal tea. Addition of milk did not alter the antioxidant activity. The influence of coffee bean source and degree of roasting was further investigated. Green coffee beans of Robusta coffee exhibited a 2-fold higher antioxidant activity than Arabica coffee, but after roasting this difference was no longer significant. In conclusion, these commonly consumed beverages have a significant antioxidant activity, the highest being soluble coffee on a cup-serving basis.     

Factors affecting the levels of tea polyphenols and caffeine in tea leaves

An isocratic HPLC procedure was developed for the simultaneous determination of caffeine and six catechins in tea samples. When 31 commercial teas extracted by boiling water or 75% ethanol were analyzed by HPLC, the levels of (-)-epigallocatechin 3-gallate (EGCG), and total catechins in teas were in the order green tea (old leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea. Tea samples extracted by 75% ethanol could yield higher levels of EGCG and total catechins. The contents of caffeine and catechins also have been measured in fresh tea leaves from the Tea Experiment Station in Wen-Shan or Taitung; the old tea leaves contain less caffeine but more EGCG and total catechins than young ones. To compare caffeine and catechins in the same tea but manufactured by different fermentation processes, the level of caffeine in different manufactured teas was in the order black tea > oolong tea > green tea > fresh tea leaf, but the levels of EGCG and total catechins were in the order green tea > oolong tea > fresh tea leaf > black tea. In addition, six commercial tea extracts were used to test the biological functions including hydroxyl radical scavenging, nitric oxide suppressing, and apoptotic effects. The pu-erh tea extracts protected the plasmid DNA from damage by the Fenton reaction as well as the control at a concentration of 100 microg/mL. The nitric oxide suppressing effect of tea extracts was in the order pu-erh tea >/= black tea > green tea > oolong tea. The induction of apoptosis by tea extract has been demonstrated by DNA fragmentation ladder and flow cytometry. It appeared that the ability of tea extracts to induce HL-60 cells apoptosis was in the order green tea > oolong > black tea > pu-erh tea. All tea extracts extracted by 75% ethanol have stronger biological functions than those extracted by boiling water.     

Effects of tea polyphenols on emulsification of olive oil in a small intestine model system

Tea catechins have been shown to reduce plasma cholesterol and suppress hypertriacylglycerolemia by reducing triglyceride absorption. However, the mechanism is not yet clear. One of the possible mechanisms is that tea polyphenols may modify dietary fat emulsification in the gastrointestinal tract. The digestive enzyme (lipase) acts on specific emulsion interface properties (droplet size and surface area). Therefore, changes in these properties may modify emulsification and lead to changes in dietary fat digestion and absorption. In this study, the effect of both green and black tea on the changes of emulsification was examined by measuring the droplet size and the surface area. A model emulsion system containing olive oil, phosphatidylcholine (PC), and bile salt was developed to simulate small intestinal conditions. Initial changes in droplet size (from 1.4 to 52.8 microm and from 1.4 to 25.9 microm) of the emulsion were observed in the presence of 1.04 mg/mL and 0.10 mg/mL of total catechins prepared from green and black tea, respectively. Both teas caused similar changes on the emulsion properties; however, black tea was more effective than green tea. The underlying mechanisms of actions of tea polyphenols are discussed.     

Antioxidative activities of phenylethanoid glycosides from Ligustrum purpurascens.

Tea and kudingcha (bitter tea) are the two most popular beverages consumed in China. Tea derived from the leaves of Camellia sinensis has been well studied for its various health benefits, but there are very limited data on the biological activities of bitter tea derived from the leaves of Ligustrum purpurascens (LP). The present study was carried out to characterize the antioxidants present in the bitter tea brewed from the leaves of LP. It was found that the crude glycoside fraction possessed strong protection against oxidation of human low-density lipoprotein (LDL). The column chromatographic separation led to the isolatation of five phenylethanoid glycosides, namely, acteoside, ligupurpuroside A, cis-ligupurpuroside B, trans-ligupurpuroside B, and osmanthuside B. When acteoside was heated in the boiling water, it was isomerized to form isoacteoside. Acteoside, isoacteoside, and ligupurpuroside A purified from LP were protective, whereas cis-ligupurpuroside B, trans-ligupurpuroside B, and osmanthuside B exhibited no protection to human LDL from Cu(2+)-medicated oxidation. Acteoside, isoacteoside, and ligupurpuroside A were also effective in preventing the peroxyl free radical-induced oxidation of alpha-tocopherol in human LDL. The antioxidant activities of acteoside, isoacteoside, and ligupurpuroside A were comparable to that observed for a green tea antioxidant, (-)-epicatechin gallate. The inhibitory effect of these three phenylethanoid glycosides on oxidation of human LDL and alpha-tocopherol was dose-dependent at concentrations of 5-40 microM. The present results suggest that the bitter tea beverage derived from LP contains effective antioxidants that may have an equal benefit as a green tea beverage.     

Determination and comparison of γ-aminobutyric acid (GABA) content in pu-erh and other types of Chinese tea

Two previous studies have reported that pu-erh tea contains a high level of γ-aminobutyric acid (GABA), which is the major inhibitory neurotransmitter in the central nervous system and has several physiological functions. However, two other researchers have demonstrated that the GABA content of several pu-erh teas was low. Due to the high value and health benefits of GABA, analysis of mass-produced pu-erh tea is necessary to determine whether it is actually enriched with GABA. A high-performance liquid chromatography (HPLC) method was developed for the determination of GABA in tea, the results of which were verified by amino acid analysis using an Amino Acid Analyzer (AAA). A total of 114 samples of various types of Chinese tea, including 62 pu-erh teas, 13 green teas, 8 oolong teas, 8 black teas, 3 white teas, 4 GABA teas, and 16 process samples from two industrial fermentations of pu-erh tea (including the raw material and the first to seventh turnings), were analyzed using HPLC. Statistical analysis demonstrated that the GABA content in pu-erh tea was significantly lower than that in other types of tea (p < 0.05) and that the GABA content decreased during industrial fermentation of pu-erh tea (p < 0.05). This mass analysis and comparison suggested GABA was not a major bioactive constituent and resolved the disagreement GABA content in pu-erh tea. In addition, the GABA content in white tea was found to be significantly higher than that in the other types of tea (p < 0.05), leading to the possibility of producing GABA-enriched white tea.     

Differentiation of tea (Camellia sinensis) varieties and their geographical origin according to their metal content

The metal content of 46 tea samples, including green, black, and instant teas, was analyzed. Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na, Sr, Ti, and Zn were determined by ICP-AES. Potassium, with an average content of 15145.4 mg kg(-1) was the metal with major content. Calcium, magnesium, and aluminum had average contents of 4252.4, 1978.2, and 1074.0 mg kg(-1), respectively. The average amount of manganese was 824.8 mg kg(-1). There were no clear differences between the metal contents of green and black teas. Pattern recognition methods such as principal component analysis (PCA), linear discriminant analysis (LDA), and artificial neural networks (ANN), were applied to differentiate the tea types. LDA and ANN provided the best results in the classification of tea varieties. These chemometric procedures were also useful for distinguishing between Asian and African teas and between the geographical origin of different Asian teas.     

Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine

Black tea, green tea, red wine, and cocoa are high in phenolic phytochemicals, among which theaflavin, epigallocatechin gallate, resveratrol, and procyanidin, respectively, have been extensively investigated due to their possible role as chemopreventive agents based on their antioxidant capacities. The present study compared the phenolic and flavonoid contents and total antioxidant capacities of cocoa, black tea, green tea, and red wine. Cocoa contained much higher levels of total phenolics (611 mg of gallic acid equivalents, GAE) and flavonoids (564 mg of epicatechin equivalents, ECE) per serving than black tea (124 mg of GAE and 34 mg of ECE, respectively), green tea (165 mg of GAE and 47 mg of ECE), and red wine (340 mg of GAE and 163 mg of ECE). Total antioxidant activities were measured using the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays and are expressed as vitamin C equivalent antioxidant capacities (VCEACs). Cocoa exhibited the highest antioxidant activity among the samples in ABTS and DPPH assays, with VCEACs of 1128 and 836 mg/serving, respectively. The relative total antioxidant capacities of the samples in both assays were as follows in decreasing order: cocoa > red wine > green tea > black tea. The total antioxidant capacities from ABTS and DPPH assays were highly correlated with phenolic content (r2 = 0.981 and 0.967, respectively) and flavonoid content (r2 = 0.949 and 0.915). These results suggest that cocoa is more beneficial to health than teas and red wine in terms of its higher antioxidant capacity.     

Phenol antioxidant quantity and quality in foods: beers and the effect of two types of beer on an animal model of atherosclerosis

The free phenols have been measured in 15 lagers, 6 porters and ales, and 11 light and nonalcoholic beers. Phenols were measured colorimetrically using an oxidation-reduction reaction with Folin-Ciocalteu reagent and catechin as the standard. The order of phenol concentration was ales > lagers > low calorie > nonalcoholic. The quality of antioxidants of the major phenols in beers and the quality of beer antioxidants were measured by (1) dose-response inhibition of lower density lipoprotein oxidation and (2) concentration of phenols in the beers at which 50% of the peroxide was destroyed in a luminescent assay for antioxidant activity. The beers' lipoprotein antioxidant quality was clearly superior to that of vitamin antioxidants and to that of the phenol ingredients, suggesting synergism among the antioxidants in the mixture. The average per capita consumption of beer in the United States in 2000 was 225 mL/day, equivalent to 42 mg/day of catechin equivalents. Beer provides more antioxidants per day than wine in the U.S. diet. A dark beer and a lager beer were given at two concentrations to cholesterol-fed hamsters, an animal model of atherosclerosis. At the high dose ((1)/(2)-diluted beer) both lager and dark beer significantly inhibited atherosclerosis compared to a control of 2% alcohol. At the high dose, lager significantly decreased cholesterol and triglycerides, and both beers acted as in vivo antioxidants by decreasing the oxidizability of lower density lipoproteins. At the low dose ((1)/(10)-diluted beer) only the lager beer significantly decreased atherosclerosis compared to the 0.4% alcohol control. The polyphenols in the beers appear to be responsible for the benefits of beer in this model. Lager beer inhibited atherosclerosis at a human equivalent dose in this hamster model of atherosclerosis.     

Massive accumulation of gallic acid and unique occurrence of myricetin, quercetin, and kaempferol in preparing old oolong tea

Old oolong tea, tasting superior and empirically considered beneficial for human health, is prepared by long-term storage accompanied with periodic drying for refinement. Analyzing infusions of three old and one newly prepared oolong teas showed that significant lower (-)-epigallocatechin gallate (EGCG) but higher gallic acid contents were detected in the old teas compared to the new one. The possibility of releasing gallic acid from EGCG in old tea preparation was supported by an in vitro observation of gallic acid degraded from EGCG under heating conditions mimicking the drying process. Moreover, three minor flavonols, myricetin, quercetin, and kaempferol, that were undetectable in the new tea occurred in all of the three old teas. Converting the new oolong tea into an old one by periodic drying revealed the same characteristic observation, i.e., massive accumulation of gallic acid presumably released from EGCG and unique occurrence of flavonols putatively decomposed from flavonol glycosides.     

Black and green teas equally inhibit diabetic cataracts in a streptozotocin-induced rat model of diabetes

Green and black teas were given at 1.25% in the drinking water to streptozotocin-induced diabetic rats for 3 months. Normal and diabetic control groups were also studied. As expected, diabetic animals had significantly increased glucose in lens and plasma. Lens and red blood cell sorbitol were significantly increased as a result of the aldose reductase pathway activation. Plasma and lens lipid thiobarbituric acid-reactive substances and protein glycation were also significantly elevated. Both teas significantly inhibited diabetic cataracts and caused significant reductions in the biochemical pathway implicated in the development of the pathology. After corrections for glucose, it was found that the teas retard the development of diabetic cataracts by a hypoglycemic effect that in turn inhibits the biochemical indicators of pathology. There were significant correlations between glucose, cataract score, and these indicators. Green tea but not black tea caused a significant decline in triglycerides in the diabetic animals. Tea may be a simple, inexpensive means of preventing or retarding human diabetes and the ensuing complications. Tea also should be investigated as an adjunct therapy for diabetes treatment.