Yongquan Xu

Improving the Effectiveness of (−)-Epigallocatechin Gallate (EGCG) against Rabbit Atherosclerosis by EGCG-Loaded Nanoparticles Prepared from Chitosan and Polyaspartic Acid

(-)-Epigallocatechin gallate (EGCG) is the major bioactive compound in green tea. Its effect is limited by the harsh environment of the gastrointestinal tract. The present study investigates how the effectiveness of EGCG is influenced by its encapsulation into self-assembled nanoparticles of chitosan (CS) and aspartic acid (PAA). Blank nanoparticles with a mean diameter of ca. 93 nm were prepared from 30-50 kDa PAA and 3-5 kDa CS with a mass rate of 1:1. EGCG was loaded in the nanoparticles to yield EGCG-CS-PAA nanoparticles with an average diameter of 102 nm, which were pH-responsive and demonstrated different EGCG release profiles in simulated gastrointestinal tract media. The average ratio (%) of lipid deposition for EGCG-CS-PAA nanoparticles administered orally to rabbits was 16.9 ± 5.8%, which was close to that of oral simvastatin (15.6 ± 4.1%). Orally administered EGCG alone yielded an average ratio of lipid deposit area of 42.1 ± 4.0%, whereas this value was 65.3 ± 10.8% for the blank nanoparticles. The effectiveness of EGCG against rabbit atherosclerosis was significantly improved by incorporating EGCG into the nanoformulation.

Improving the sweet aftertaste of green tea infusion with tannase

The present study aims to improve the sweet aftertaste and overall acceptability of green tea infusion by hydrolyzing (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) with tannase. The results showed that the intensity of the sweet aftertaste and the score of overall acceptability of the green tea infusion significantly increased with the extension of the hydrolyzing treatment. (-)-Epigallocatechin (EGC) and (-)-epicatechin (EC) were found to be the main contributors for the sweet aftertaste, based on a trial compatibility with EGCG, ECG, EGC, and EC monomers, and a synergistic action between EGC and EC to sweet aftertaste was observed. A 2.5:1 (EGC/EC) ratio with a total concentration of 3.5 mmol/L gave the most satisfying sweet aftertaste, and the astringency significantly inhibited the development of the sweet aftertaste. These results can help us to produce a tea beverage with excellent sweet aftertaste by hydrolyzing the green tea infusion with tannase.

The impact of Ca2+ combination with organic acids on green tea infusions

The effect of Ca(2+) in brewing water on the organic acid content, turbidity, and formation of tea cream and sediment in green tea infusions was studied. When the Ca(2+) concentration of the brewing water was >40 mg L(-1), the green tea infusion became more turbid. The turbidity of the tea infusion was highly negatively correlated with the contents of oxalic acid (R=-0.89, p<0.01), quinic acid (R=-0.90, p<0.01) and tartaric acid (R=-0.82, p<0.01). Oxalic acid on its own interacted with Ca(2+) at low concentrations, whereas polyphenols and protein did not. In conclusion, Ca(2+) in brewing water influences the quality of a tea infusion by inducing tea cream and sediment formation from combination of Ca(2+) and organic acids, such as oxalic acid, quinic acid and tartaric acid. Ca(2+) and oxalate are the main metal ion and anion, respectively, involved in tea cream and sediment formation on tea infusion cooling or concentrating.

Analysis of cream formation in green tea concentrates with different solid concentrations

The formation of tea cream in the green tea concentrates of different solid concentrations (5, 10, 20, 30, 40, 50 and 60°Brix) was investigated. The results showed a good positive correlation (γ = 0.98, p ≤ 0.05) between the amount of tea cream and the solid concentrations from 5 to 40°Brix, while the amount of tea cream in the tea concentrates of 50 and 60°Brix decreased acutely. Total sugar, caffeine and catechins were found to be the main chemical components of tea cream in the green tea concentrate. The large decrease of the amount of tea cream in the tea concentrates of 50 and 60°Brix may be induced by a sharp increase of the viscosity of the tea concentrates, which helped to improve the stability of tea concentrate. It may be indicated that the stability of green tea concentrate enhanced when the concentration higher than 50°Brix, which helped to restrain the formation of tea cream.

The major factors influencing the formation of sediments in reconstituted green tea infusion

The effects of Ca(2+), caffeine and polyphenols on the formation of reversible tea sediments (RTS) and irreversible tea sediments (IRS) in green tea infusion were studied. Adding Ca(2+) (2 mmol/l) was found to increase the formation of RTS by 8% and IRS by 92%, while adding chelating ions of Na2EDTA significantly decreased the amount of RTS by 14.6%, but not the amount of IRS. Under acid conditions, Ca(2+) combined with oxalic ions to form indissoluble oxalate that is the principal constituent of IRS, despite the existence of the chelating ions. Decaffeination largely inhibited the formation of RTS (73%) and IRS (60%), even in the presence of Ca(2+). The amount of sediment could be reduced by removing polyphenols using polyvinyl-polypyrrolidone. The results suggest that sediment formation in green tea infusions can be inhibited by lowering the concentration of Ca(2+), caffeine or polyphenols.

Scaling up of high-speed countercurrent chromatographic apparatus with three columns connected in series for rapid preparation of (−)-epicatechin

In the present study, compact high-speed countercurrent chromatographic apparatus was constructed with three columns connected in series. Two sets of columns were prepared from 10 mm and 12 mm I.D. tubing to form 12 L and 15 L capacities, respectively. Performance of these columns was compared for the separation of (-)-epicatechin gallate (ECG) from a tea extract by flash countercurrent chromatography (FCCC). In each separation, 200 g of the tea extract in 1600 mL of mobile phase was loaded onto the column. The 12 L column gave 7.5 L (35 g of ECG) and the 15 L column gave 9 L (40 g of ECG) of ECG solution without impurities. The ECG solution was directly hydrolyzed by tannase into (-)-epicatechin. The hydrolysate was purified by flash chromatography on AB-8 macroporous resin to give 52 g of EC (purity 99.1%). This scaled up apparatus could be used for the industrial separation of natural products.

Irreversible Sediment Formation in Green Tea Infusions

The formation of irreversible tea sediment (IRS) and its chemical components in green tea infusions were investigated. The results showed that the amounts of IRS in the green tea infusions from various tea cultivars ranged from 0.10 to 1.47 mg/mL. The amount of IRS was influenced remarkably by the chemical components in the green tea infusion. Principal component analysis and regression analysis indicated that gallated catechins, Mn, Ca, caffeine, Na, and (-)-gallocatechin gallate (GCG) were the principal components. IRS (mg/mL) = -4.226 + 0.275 gallated catechins + 79.551 Na + 7.321 Mn + 21.055 Ca + 0.513 caffeine - 0.129 GCG (R2 = 0.697). The contents of the main chemical components in the reversible tea sediment (RTS) and IRS were markedly different, especially the minerals. Large amount of minerals participated in the formation of irreversible green tea sediment. The amount of IRS increased with the extraction temperature.

Sediments in concentrated green tea during low-temperature storage

The formation and the main chemical components of sediments, including reversible tea sediments (RTS) and irreversible tea sediments (IRS), in concentrated green tea during low-temperature storage were studied. RTS was mainly formed in the first 10 days, and IRS was mainly formed between 20 and 40 days of storage. The RTS were the primary sediment, contributing more than 90% of the total sediment. The RTS comprised of polyphenols, total sugar, caffeine, flavones and proteins, while the IRS mainly comprised of oxalates of Ca, Mg, Ga and Mn. The total mineral content in the IRS (17.1%) was much higher than that in the RTS (2.6%) after 80 days of storage. The Ca, Mg, Mn and Ga contents in IRS were over 1.0% (w/w) each. About 75% of the IRS was soluble in 0.1 M aqueous HCl, with the oxalate accounting for 68%. Minerals and oxalic acid were the crucial factors in the IRS formation.

Effect of extraction methods on the chemical components and taste quality of green tea extract

The physicochemical properties of tea extracts are significantly affected by the extraction method. The aim of this study was to compare the effects of static and dynamic extractions on the concentrations of chemical components and taste quality of green tea extracts. Our results show that extraction of chemical components using static extraction follows a pseudo-second-order reaction, while that of dynamic extraction follows a first-order reaction. The concentrations of the solids, polyphenols, and free amino acids in green tea extract prepared by dynamic extraction were much higher, although the overall yields were not significantly different between the two extraction methods. Green tea extracts obtained via dynamic extraction were of lower bitterness and astringency, as well and higher intensities of umami and overall acceptability. These results suggest that dynamic extraction is more suitable for the processing of green tea concentrate because of the higher concentration of green tea extract.

Quantitative analyses of the bitterness and astringency of catechins from green tea

Bitterness and astringency are two important quality attributes of green tea infusion, and catechins are the main contributor to the bitterness and astringency. The aim of this work was to quantitatively analyse the bitterness and astringency of green tea infusion according to the concentrations of catechins. The concentration-taste curves of catechins showed a pattern that fit the cubic functions, and their R2 values were higher than 0.956. The bitterness of green tea was highly correlated with the concentrations of (-)-epigallocatechin gallate and (-)-epicatechin gallate (ECG) (R2 = 0.7769, p < 0.01), and the astringency (R2 = 0.7878, p < 0.01) was highly correlated with the concentrations of ECG and flavonol glycosides (myricetin 3-O-galactoside and quercetin-3-O-rutinoside). Taste interactions between different catechins and between catechins and other substances were determined. These results may enhance the understanding of tea chemistry for improving the taste of products from green tea.