ChapterPDF Available

Green Tea (Cv. Benifuuki) Powder and Catechins Availability

Authors:

Figures

Content may be subject to copyright.
Green tea (cv. Benifuuki) powder and catechins availability
Mari Maeda-Yamamoto [1]
Hirofumi Tachibana [2]
Yoichi Sameshima [3]
Shinichi Kuriyama[4]
[1] Corresponding author. Mari Maeda-Yamamoto: Institute of Vegetable and
Tea Science, National Agriculture and Food Research Organization, Kanaya
2769, Shimada, Shizuoka 428-8501, Japan. Telephone: +81 547(45)4101,
FAX: +81 547(46)2169, Email:<marimy@affrc.go.jp>
[2] Associate professor Tachibana: Bioscience & Biotechnology Faculty of
Agriculture, Graduate School of Bioenvironmental Sciences, Kyushu
University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan.
Telephone/FAX: +81 92(642)3008, Email:<tatibana@agr.kyushu-u.ac.jp>
[3] MD. Sameshima: Kakegawa Municipal General Hospital, Sugiya-minami
1-1-1, Kakegawa, Shizuoka 436-8502, Japan. Telephone: +81 537(22)6211,
FAX: +81 537(24)2539, Email:<mdysame@yahoo.co.jp>
[4] Professor MD.Kuriyama: Department of Molecular Epidemiology,
Environment and Genome Research Center, Tohoku University Graduate
School of Medicine, Seiryo-cho 2-1, Aoba-ku, Sendai, Miyagi 980-8575,
Japan. Telehone: +81 22(717)8950, FAX: +81 22(717)8951, Email:
<Kuriyama@med.tohoku.ac.jp>
Running title/ Green tea powder and catechins availability
Word count (excluding facing page, abstract, and references)
3069 words, 3 tables, 4 figures. 30 references
Summary
Benifuuki, a tea cultivar grown in Japan, is rich in the anti-allergic molecule
epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3”Me). This study was
conducted to investigate the absorption of catechins in rats and human after the
oral administration of Benifuuki green tea (tea powder or with foodstuffs
increased in functionality). The absorption of EGCG3Me for the pulverized
Benifuuki green tea powder (BGP) 2.86µm (mean particle size) and the infusion
were significantly higher than those in BGP 18.6 µm and 76.1µm. The
absorption of EGCG and ECG for BGP 2.86µm were significantly higher than
those in the infusion and 18.6µm and 76.1µm BGP, but the absorption of EGC
and EC showed no differences between these groups in rats. In EGCG of
human, the AUC of BGP 2.86µm with milk tended to be higher than BGP, and
vitamin A or vitamin E had a tendency to increase the AUC of BGP with milk.
Key Words
epigallocatechin-3-O-(3-O-methyl) gallate, ester-type catechins, free-type
catechins, Benifuuki green tea, absorption, tea powder, anti-allergic action
Abbreviations
EGCG3Me; epigallocatechin-3-O-(3-O-methyl)-gallate, EGCG;
epigallocatechin-3-O-gallate, ECG; epicatechin-3-O-gallate, EGC;
epigallocatechin, EC; epicatechin, BGP; Benifuuki green tea powder, BGI;
Benifuuki green tea infusion, AUC; the area under the plasma concentration
-time curve,
Introduction
Tea (Camellia sinensis L.) is consumed all over the world, particularly in Japan
and China, where it has been used for medicinal purposes for tens of centuries.
It has been reported that tea has various bioregulatory activities. Catechin, a
group of polyphenolic compounds, have been shown to be largely responsible
for these activities. Epigallocatechin-3-O-gallate (EGCG) is the major catechin
component of green tea and may be its major active constituent.
Allergies are defined as diseases that provoke excessive immune activity
(Kinet, 1999; Kawakami and Galli. 2002), and in Japan, allergy morbidity is
estimated to be about 30%. Many Japanese people have misgivings about the
use of anti-allergic medicine because of the side effects and expense so there is a
demand for physiologically-functional foods for allergy prevention to be
developed. Catechins have been demonstarted to have anti-allergic effects
(Matsuo, 1997; Yamashita, 2000). We have previously demonstarted that
O-methylated EGCG (epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3”Me),
epigallocatechin-3-O-(4-O-methyl) gallate (EGCG4”Me) (Sano,1999; Suzuki,
2000; Fujimura, 2002; Maeda-Yamamoto, 2004; Fujimura, 2007), and strictinin
(Tachibana et al. 2001) have anti-allergic actions and that the Japanese tea
cultivar Benifuuki is rich in EGCG3”Me. EGCG3”Me disappeared in black
tea by polyphenol oxidase and the content was high in fully-matured tea leaves
late in the first crop season or under the third leaf during the second crop season
(Maeda-Yamamoto, 1998; Maeda-Yamamoto, 2001; Maeda-Yamamoto, 2004).
The oral administration of these methylated catechins significantly and
dose-dependently (5–50 mg/kg) inhibited type I allergic (anaphylactic) reactions
in mice sensitized with ovalbumin and Freunds incomplete adjuvant. These
catechins also strongly inhibited mast cell activation by preventing the tyrosine
phosphorylation (Lyn, Syk, and Btk) of cellular proteins, the myosin light chain
phosphorylation via the suppression of 67LR expression (Fujimuara, 2007),
histamine/leukotriene release, and interleukin-2 secretion after Fcepsilon RI
cross-linking (Maeda-Yamamoto, 2004) .
Previously, the bioavailability of EGCG3”Me and EGCG after the oral
administration of Benifuuki green tea infusion in humans was reported
(Maeda-Yamamoto, 2007). Pulverized Benifuuki green tea powder (BGP) is
more widely-distributed than leaf tea in Japan. Japanese people mix pulverized
tea with water directly, whereas it is common to drink leaf tea after extraction.
However, few studies of the effects of BGP particle size or coexisting foodstuffs
on catechin bioavailability have been performed. In this manuscript, we
examine the blood levels of EGCG3Me and other catechins after oral
administration of pulverized green tea (cv. Benifuuki) powders with different
particle sizes, infused tea or tea powder with foodstuffs to rats or humans.
Bioavailability of Benifuuki green tea infusion and anti-allergic activity of the
tea in humans
After oral administration of Benifuuki green tea infusion containing 43.5mg
of EGCG and 8.5mg of EGCG3”Me, the AUC (the area under the plasma
concentration-time curve; min·µg/ml) of EGCG was 6.72±2.87 and EGCG3"Me
was 8.48±2.54 in ten healthy human volunteers, as shown in Figure 1. Though
the dose of EGCG was 5.1 times the dose of EGCG3"Me, the AUC of
EGCG3Me was higher than that of EGCG (Maeda-Yamamoto, 2007). It is
suggested that EGCG3Me was absorbed easier than EGCG and metabolic
clearance rate of EGCG3Me was slower than EGCG, and these actions were
related to strong in vivo allergic effects.
We investigated the in vivo effect of Benifuuki green tea (extract) on the
symptom relief and safety in subjects with seasonal allergic rhinitis. A
double-blind clinical study on subjects with Japanese cedar pollinosis was
carried out. At the eleventh week after starting the study, in the most severe
cedar pollen scattering period, symptoms, i.e. blowing the nose and itching eyes,
were significantly relieved in the Benifuuki green tea intake group compared
with the placebo group (Yabukita green tea (extract) did not contain
O-methylated catechins and was used as placebo), and blowing the nose, itching
eyes and nose symptom score, and at the eleventh and thirteenth weeks, stuffy
nose, throat pain and the nose symptom medication score were significantly
relieved in the Benifuuki containing ginger extract group compared with the
placebo group (Figure 2). These results suggested that over one consecutive
month, drinking Benifuuki green tea was useful to reduce some of the symptoms
from Japanese cedar pollinosis, and did not affect any normal immune response
in subjects with seasonal rhinitis, and the ginger extract enhanced the effect of
Benifuuki green tea (Maeda-Yamamoto, 2007).
Furthermore, we evaluated the efficacy and safety of Benifuuki green tea
containing O-methylated catechin in patients with mild perennial allergic rhinitis.
Seventy-five patients with mild perennial allergic rhinitis meeting the
predetermined criteria for subjects were assigned to either Benifuuki green tea
beverage group or Yabukita green tea beverage group. The subjects took 700 mL of
tea beverage (34mg of O-methylated catechin contain in 700mL) and recorded
their nasal and ocular symptoms every day for 12 weeks, and visited hospital every
6 weeks for consultation and blood collection. As a result, the scores for nasal
and ocular symptoms of Benifuuki group were found to be lower than those of
Yabukita with statically significances at 7th-12th week for nasal scores and at
4th-12th week for ocular scores (Yasue, 2005). No adverse effect was observed
in physiological, hematological, biochemical parameters, normal immune
responses of peripheral blood leukocytes, and subjective symptom throughout the
experiment. An additional study in 9 healthy subjects without any allergic
symptoms was also conducted. The subjects were daily given 700 ml of Benifuuki
green tea for 12 weeks, and no adverse effect was noted throughout the study.
These results suggest that Benifuuki green tea beverage containing O-methylated
catechin is useful for the treatment of mild perennial allergic rhinitis.
Moreover, when 27 of atopic dermatitis patients (moderate) swabbed
Benifuuki tea extract containing cream to affected part for 8 weeks, the
symptom significantly improved and the amount of consumption of
corticosteroid significantly decreased compared with base cream (Fujisawa,
2005).
Pharmacokinetics of plasma catechins after the intragastric administration of
Benifuuki green tea in rats
To study the bioavailability of EGCG3”Me and other catechins after the
administration of green tea, Benifuuki green tea powder (BGP) with different
particle sizes (2.86 µm (BGP 2.86 µm), 18.6 µm (BGP 18.6 µm), and 76.1 µm
(BGP 76.1 µm)) or Benifuuki green tea infusion (BGI; control; 1.25 g of
Benifuuki green tea were extracted for 10 min in 20ml of boiled distilled water)
was given to rats i.g. After the Benifuuki green tea had been i.g. administered
to the rats at 125 mg/head equivalency, blood samples were collected at different
time points (1, 3, 6, 11, and 24 h). Table 1 shows a comparison of intake,
Tmax (maximum drug concentration time), Cmax (maximum drug
concentration) and AUC for plasma catechins after the intragastric
administration of the Benifuuki green teas (3 powders with different particle
sizes and an infusion) (Maeda-Yamamoto, 2011). The intragastric
administration of the green teas (BGP and BGI) did not result in significant
changes in the Tmax of any catechin (1.17 ± 0.09 - 2.89 ± 1.89 h). In the BGP
2.86µm group, the Cmax of ester-type catechins (free EGCG3Me, total EGCG,
and total ECG) were significantly higher than those of the other groups (BGP
2.86 µm > BGI > BGP 76.4 µm = BGP 18.6 µm).
Lambert et al (2003b) demonstrated that in a comparison of the EGCG
concentration of sulfatase / glucuronidase-treated plasma (total EGCG) with that
of free plasma (unconjugated EGCG), 50 - 90% of EGCG was present in the
conjugated form in mouse plasma after oral administration of EGCG. Lee et al
(2002) indicated that after 1 h ingestion, 77% of the EGCG was present in the
free (unconjugated) form whereas 31% of EGC and 21% of EC were in the free
form. O-methylated EGCG was one of metabolite. So, EGCG, ECG, EGC
and EC were shown as total (sulfatase / glucuronidase -treated) catechin.
The mean intake concentrations of ester-type catechins in the BGI group were
about half of those in the other BGP groups, and the differences between BGI
group and the other BGP groups were significant.
Furthermore, the AUC of ester-type catechins in the BGP 2.86 µm group
were significantly higher than those of the other groups (approximately 2-3 fold
higher).
Absorption of catechins after intragastric administration in rats
Figure 3 shows the mean absorption of EGCG3Me (A), EGCG (B), ECG (C),
BGP 2.86µm BGP 18.6µm BGP 76.1µm BGI
EGCG3"Me
In tak e (m g /ra t)
1.88 ± 0.05
a
1.81 ± 0.04
ab
1.74 ± 0.02
b
0.84 ± 0.04
c
T
max
(h) 1.57 ± 0.58 1.17 ± 0.09 1.85 ± 0.95 1.47 ± 0.10
C
max
(µg/L) 224.8 ± 98.9
a
40.5 ± 47.3
b
64.8 ± 40.6
b
74.5 ± 24.3
b
AUC (µg•h/L)
760.9 ± 273.4
a
229.9 ± 152.3
b
319.8±123.7
b
288.2 ± 112.6
b
EGCG
In tak e (m g /ra t)
7.71 ± 0.15
a
7.46 ± 0.06
a
7.41 ± 0.05
a
3.60 ± 0.17
b
T
max
(h) 1.74 ± 0.57 2.89 ± 1.89 1.85 ± 0.88 2.76 ± 0.75
C
max
(µg/L) 642.7 ± 263.6
a
148.3 ± 97.1
b
739.3 ± 275.2
a
130.9 ± 63.4
b
AUC (µg•h/L)
2763.5 ± 1158.9
a
968.3 ± 169.6
b
1157.7 ± 417.7
b
1166.4 ± 312.8
b
ECG
In tak e (m g /ra t)
1.95 ± 0.04
a
1.88 ± 0.02
a
1.87 ± 0.01
a
0.80 ± 0.04
b
T
max
(h) 1.42 ± 0.09 2.87 ± 2.04 1.18 ± 0.62 2.11 ±1.08
C
max
(µg/L) 171.2 ± 82.6
a
44.2 ± 34.2
b
79.4 ± 61.6
a
58.0 ± 24.5
b
AUC (µg•h/L)
660.8 ± 191.9
a
305.2 ± 151.2
b
315.4 ± 150.9
b
222.1 ± 87.2
b
EGC
In tak e (m g /ra t)
4.42 ± 0.09
a
4.30 ± 0.02
a
4.31 ± 0.04
a
3.75 ± 0.16
b
T
max
(h) 1.86 ± 0.61 1.55 ± 0.27 1.83 ± 0.93 1.47 ± 0.27
C
max
(µg/L) 903.1 ± 202.0 819.0 ± 251.0 757.6 ± 247.1 674.4 ± 213.1
AUC (µg•h/L) 3577.0 ± 793.2 3447.5 ± 841.5 2887.0 ± 923.2 2463.2 ± 730.1
EC
In tak e (m g /ra t)
1.38 ± 0.03
a
1.34 ± 0.02
a
1.33 ± 0.02
a
1.16 ± 0.05
b
T
max
(h) 1.83 ± 0.66 1.78 ± 0.75 1.83 ± 0.88 1.45 ± 0.28
C
max
(µg/L) 420.8 ± 148.2 571.2 ± 299.5 405.5 ± 168.3 479.9 ± 174.7
AUC (µg•h/L) 1288.3 ± 315.7 1300.8 ± 462.3 1192.6 ± 839.9 1091.7 ± 443.0
Table 1. Pharmacokinetic parameters of plasma catechins after i.g.
administration of Benifuuki green tea in rats
All values represent the mean ± SD of 5 mesurements except for the intake values (3
measurements). BGP; Benifuuki green tea powder, BGI; Benifuuki green tea infusion.
In each row, the means with different superscript letters are significantly different, P<0.05
(ANOVA and Tukey-Kramer's method).
EGC (D), and EC (E) concentration-time profiles after Benifuuki green tea
administration (Maeda-Yamamoto, 2011). The peak absorption levels of EGC,
EGCG, EC, EGCG3”Me, and ECG were 0.13 ± 0.06%, 0.08 ± 0.01%, 0.1 ±
0.05%, 0.21 ± 0.06%, and 0.32 ± 0.15%, respectively. The EGCG3Me
absorption rates of BGP 2.86 µm, 18.6 µm, 76.4 µm, and BGI were 0.52%,
0.16%, 0.23%, and 0.42%, respectively. The EGCG absorption rates of BGP
2.86 µm, 18.6 µm, 76.4 µm, and BGI were 0.46%, 0.16%, 0.19%, and 0.39%,
respectively. The ECG absorption rates of BGP 2.86 µm, 18.6 µm, 76.4 µm,
and BGI were 0.43%, 0.20%, 0.21%, and 0.40%, respectively. The EGC
absorption rates of BGP 2.86 µm, 18.6 µm, 76.4 µm, and BGI were 1.05%,
0.96%, 0.82%, and 0.80%, respectively. The EC absorption rates of BGP 2.86
µm, 18.6 µm, 76.4 µm, and BGI were 1.21%, 1.20%, 1.12%, and 1.17%,
respectively. Similar to the AUC of the ester-type catechins, the absorptions
of ester-type catechins were highest in the BGP 2.86µm group. However,
among the tested groups, the peak absorption level of EGCG3Me was highest
in the BGP 2.86µm and BGI groups (BGP 2.86 µm = BGI > BGP 76.4 µm =
BGP 18.4 µm). The peak absorption levels of free-type catechins (EGC and
EC) did not differ between the groups.
The plasma levels of ester-type catechins (EGCG3Me, EGCG, ECG) were
highest in the BGP 2.86 µm, but those of free-type catechins (EGC and EC)
showed no difference among the 4 groups. The absorption of free-type
catechins in rats was higher than that of ester-type catechins. In ester-type
catechins, absorption was highest in the BGP 2.86 µm group, followed by the
BGI, BGP 18.6 µm, and BGP 76.4 µm groups. This result suggested that a
Benifuuki green tea powder particle size of around 2 µm would be good for
efficiently delivering the anti-allergic EGCG3Me or the anti-oxidant EGCG.
The mean particle size of Matcha green tea, which is commonly used, was
reported to be approximately 20 µm (Haraguchi, 2003, Sawamura, 2010).
However, in this study the ester-type catechin plasma level was higher in the
BGP 2.86 µm group than in the infusion or larger particle size groups. The
present study demonstrated that the bioavailability of the beneficial components
of Benifuuki green tea, especially ester-type catechins, could be significantly
improved by reducing the particle size (11 to 2.8 µm). Li et al. (2008)
demonstrated that reducing the particle size (3.5 µm to 220 nm) produced
significant increases (two-fold) in the antioxidant and antitumor activities of
green tea particles in vitro. Deng et al. demonstrated that realgar with a smaller
particle size of 150 or 100 nm markedly inhibited cell viability through
apoptosis compared with that with a particle size of 200 nm or 500 nm. Suzuki
et al. (2003) demonstrated that synthesized catechins were localized to restricted
regions within the large central vacuoles (5 - 15 µm) or some small vacuoles
(0.5 - 3 µm) in tea leaf mesophyll cells. It is conceivable that catechin form
complexes with metal ions such as Ca (II) and Mg (II) and special proteins in
central vacuole. Suzuki et al. (2003) supposed that synthesized catechins in ER
or Golgi apparatus are packed in the formation of a small vacuole and small
vacuoles fuse with each other and as a result, catechins are transported into the
large central vacuole. However, localization of individual catechin was not
fully elucidated. We supposed that ester type catechins were absorbed at 2.86
µm well by these catechins being located in small vacuole mainly. That is, we
surmised that the absorption rate of ester-type catechins might have risen by
crushing the organization to size of small vacuole (approximately 2 µm) finely
and ester-type catechins were easy to extract from small vacuole.
On the other hand, Lambert et al. demonstrated that genistein (2008) from
soybean and piperine (2004) from black pepper enhanced EGCG availability.
This study revealed the modulation of EGCG bioavailability by a second dietary
component and the plasma concentrations of free-type catechin, EGC, and EC,
and there were no differences between any of the groups.
In our study, we found that the absorption of the beneficial components of
Benifuuki green tea, especially the absorption of ester-type catechins, was
significantly improved by reducing the particle size. It was previously shown
that an extremely small size particle (e.g. nano-scale powder) showed
nano-specific toxicological actions (O’brien and Cummins. 2010), so it is
important to clarify the optimal particle size and food components for
modulating bioavailability in humans in future.
The AUC of catechins after oral administration in humans
To understand the health effects of this tea in humans, we are studying the
pharmacokinetics of Benifuuki catechins in human volunteers. Six healthy
volunteers (four women, two men) participated in the study after having given
written informed consent. Their mean ± SD age was 43 ± 5 years (range 36-49
years). The study protocol was approved by the Institutional Review Board of
the National Institute of Vegetable and Tea Science (NIVTS) for Human
Research and was carried out in accordance with the Declaration of Helsinki.
The subjects were collected blood samples at prior to and 0.5, 1, 3, 6, 12 h after
each test drink ingestion, following an overnight fast. Test drink groups were
(1) BGP 2.86 µm (2g) with water, (2) BGP 2.86 µm with milk (200ml), (3) BGP
2.86 µm with milk and vitamin A (1mg), (4) BGP 2.86 µm with milk and
vitamin E (5mg), (5) BGI, and (6) BGI plus milk . Table 2 shows the mean AUC
± SD of EGCG3Me, EGCG, ECG, EGC, and EC. The AUC of ester-type or
free-type catechins had no significant difference between the groups. In
ester-type catechins, the AUC of BGI plus milk tended to be higher than that of
BGI. Moreover, in all catechin, the AUC of BGP with milk tended to be
higher than that of BGP.
Table 3 shows the mean absorption rate (%) ± SD of EGCG3Me, EGCG,
ECG, EGC, and EC after 1hr ingestion. The absorption of ester-type or
free-type catechins had no significant difference between the groups. In EGCG,
the absorption of BGP/BGI plus milk tended to be higher than that of BGP/BGI.
te s t d ri n k EGCG3"Me EGCG ECG EGC EC
(1) BGP 2.86 µm with water 112.37±30. 56 365.83±118.02 105.86±31.97 240.91±187.18 90.43±68. 28
(2) BGP 2.86 µm with milk 153.82±51. 73 407.95±98.91 151.56±53.53 285.41±57.27 94.95±22. 48
(3) BGP 2.86 µm with milk plus V.A. 113.00±54.65 406.87±246.81 130.39±75.61 185.16±42.54 65.98±23.48
(4) BGP 2.86 µm with milk plus V.E. 138.00±114.47 344. 67±218.05 108.34±47.92 161.81±36.64 51.32±19.54
(5) BGI 101.77±30. 04 453.69±101.63 157.11±39.77 203.34±23. 87 71.79±40.68
(6) BG I plus milk 128.89±63. 54 557.40±218.68 196.59±54.10 182.12±113.19 62.09±29. 13
BGP; Benifuuki green tea powder, BGI; Benifuuki green tea infusion
Table 2. AUC (the area plasma concentration-time curve) for catechins after oral administration of
various types of Benifuuki green tea in humans
AUC(µg•h/L)
These findings elucidated that we might add milk and drink if we would be
hard to drink green tea.
Consumption and usage of the green tea powder in Japan
Recently, in Japan, the consumption of the powdered green tea is increasing.
It is thought to be due to the proposal that eating six grams of green tea every
day made us healthy by Dr. Kuwano (1994) and the increasing of housewives
who were too lazy to use a teapot with green tea leaves. There are not the
statistics of the national production of whole green tea powder, but the national
production of matcha for processing is supposed with a little over 2,500t a year
(Sawamura, 2010).
What merits there will be for us to take whole green tea leaves? The green
tea includes 60-70% of insoluble ingredients (liposoluble vitamins,
water-insoluble dietary fibers, chlorophylls, proteins, and so on) and 30-40% of
water-soluble ingredients (polyphenols, caffeine, amino acids, water-soluble
vitamins, water-soluble dietary fibers, saponin, minerals, and so on). We
usually drink it as extract when we drink green tea, so all the insoluble
ingredients are disposed of as used tea leaves. Dr. Kuwano has reported that
liver vitamin A activity of vitamin A-deficient rats significantly increased by the
administration of green tea powder compared to rats that were administered
control diet without green tea powder. We need to do to ingest 22% of vitamin
A, 10% of vitamin B2, 17% of niacin, 5% of magnesium/calcium, 6% of
te s t d rin k EGCG3"Me EGCG ECG EGC EC
(1) BGP 2.86 µm with water 0.21±0.18 0.16±0.33 0. 28±0.20 0.39±0.19 0.30±0.11
(2) BGP 2.86 µm with milk 0.12±0.08 0.24±0.17 0.29±0.12 0.22±0.16 0.21±0.05
(3) BGP 2.86 µm with milk plus V.A. 0.20±0.18 0.21±0.17 0.30±0.17 0.23±0.06 0.21±0.06
(4) BGP 2.86 µm with milk plus V.E. 0.16±0.12 0.22±0.12 0.26±0.16 0.35±0.23 0.27±0.25
(5) BGI 0.22±0.12 0.16±0.13 0.25±0.18 0.53±0.26 0.41±0.12
(6) BG I plus m ilk 0.24±0.33 0.20±0.38 0.42±0.31 0.25±0.23 0.21±0.11
BGP; Benifuuki green tea pow der, BGI; Benifuuki green tea infusion
Abs orption wa s calcula ted as follows ;
Absorption = ( plasma concentration (µg/L) /1000 * blood volume (weight(kg) /14)) /(intake (mg) *1000)*100 )
Table 3. Absorption for catechins after 1hr oral administration of various types of Benifuuki green tea in humans
potassium to daily requirement, and over 2g of dietary fibers by an intake of 6g
of the green tea powder a day.
The green tea powder is made by pulverizing refined tea with a grinder mill
finely. For a crusher, there are a stone-mill, bal-mills (Figure 4), a jet-mills and
so on. It is important to process it at low temperature not to lose quality. In
addition, when we dissolve green tea powder in hot water and drink it, large and
small of the particle size of powder has a huge effect on the feeling on the throat.
If particle size is small less than 20µm, the feeling on the throat becomes good
(Sawamura, 2010; Kobayashi, 2008).
In Japan, green tea powder is sold in a bulk (aluminum bag) or on an
individual package (aluminum stick pack). It is enclosed 30-100g of tea
powder in bulk bag or 0.4-2.2g in aluminum stick pack. Benifuuki green tea
described above is often sold as powdered tea.
Basically, we stir to dissolve green tea powder in water and hot water and just
drink it. In addition, we sprinkle the green tea powder over foods (rice, a
grilled fish, fried chicken, a steak, noodles, a bowl, fermented soybeans, yogurt,
and vinegared food), mix it with foods including mayonnaise, tartar sauce, a
potato salad, a rice cake, a shao-mai, the tempura, a cake, bread, a hamburger
steak, a meatball, and process it into food boiled in soy sauce or jam.
Summary points
The investigation into changes in catechins concentrations after oral
administration of Benifuuki green tea in rats or humans elucidated the
following;
EGCG3Me was absorbed easier than EGCG and metabolic clearance rate of
EGCG3Me was slower than EGCG
The drinking Benifuuki green tea with a particle size of around 2 µm would
deliver the anti-allergic EGCG3”Me and the anti-oxidant EGCG efficiently
•The absorptions of ester-type catechins (EGCG and ECG) were highest in the
Benifuuki green tea powder (BGP) (2.86µm) group
•The peak absorption level of EGCG3Me was highest in the BGP 2.86µm and
BGI groups (BGP 2.86 µm = BGI > BGP 76.4 µm = BGP 18.4 µm)
The peak absorption levels of free-type catechins (EGC and EC) did not differ
between the groups
In ester-type catechins, the AUC of BGI plus milk tended to be higher than
that of BGI, and in all catechin, the AUC of BGP with milk tended to be higher
than that of BGP
By just drinking with green tea as powder or mixing the powdered tea in with
our foods, we can ingest the ingredients of the tea which don't dissolve in water
References
Fujimura, Y., Tachibana, H., Maeda-Yamamoto, M., Miyase, T., Sano, M.,
Yamada, K. (2002). Antiallergic Tea Catechin:
(-)-Epigallocatechin-3-O-(3-O-methyl)-gallate, Suppresses FcepsilonRI
Expression in Human Basophilic KU812 Cells. J Agric Food Chem. 50,
5729-5730.
Fujimura, Y., Umeda, D., Yano, S., Maeda-Yamamoto, M., Yamada, K.,
Tachibana, H. (2007). The 67kDa laminin receptor as a primary determinant
of anti-allergic effects of O-methylated EGCG. Biochem Biophys Res
Commun. 364,79-85.
Fujisawa, T., Horikawa, T., Adachi, Y., Murakami, K., Goto, S.(2005) Japanese
Journal of Allergology (Japanese) 54(8/9), 1022.
Galli SJ, Maurer M, Lantz CS (1999) Mast cells as sentinels of innate immunity.
Curr Opin Immuno.l 11, 53-59
Haraguchi, Y., Imada, Y., Sawamura, S. (2003). Production of characterization
of fine matcha for processed food. Nippon Shokuhin Kagaku Kougaku Kaishi
(Japanese) 50, 468-473.
Kawakami, T. and Galli, S.J. (2002). Regulation of mast-cell and basophil
function and survival by IgE. Nat Rev Immunol. 2, 773-786.
Kinet, J.P. (1999). The high-affinity IgE receptor (Fc epsilon RI): from
physiology to pathology. Annu Rev Immunol. 17, 931-972.
Kobayashi, T., Goto, T., Oomiya, T., Matsuhita, H. (2008). Relations of a
particle size and the preference of powdered green tea by the difference of the
crushing method. Tea Research Journal(Japanese) 106 supplement, 74-75.
Kuwano, K., Sakitamaki, C., Suzuki J., Mitamura, T. (1994). Efficacy of green
tea carotene as vitamin A activity in rats. Tokyo Kasei Gakuin University Kiyo
(Japanese) 34, 5-8.
Lambert, J.D. and Yang, C.S. (2003a). Cancer chemopreventive activity and
bioavailability of tea and tea polyphenols. Mutat Res. 523, 201-208.
Lambert, J.D., Lee, A.J., Lu, H., Meng, X., Hong, J.J., Seril, D.N., Sturgill.,
M.G., Yang, C.S. (2003b). Epigallocatechin-3-gallate is absorbed but
extensively glucuronidated following oral administration to mice. J Nutr.
133,4172-4177.
Lambert, J.D., Hong, J., Kim, D.H., Misin, V.M., Yang, C.S. (2004). Piperine
enhances the bioavailabiliy of the tea polyphenol
(-)-epigallocatechin-3-gallate in mice. J Nutr. 134, 1948-1952.
Lambert, J.D., Kwon, S.J., Ju, J., Bose, M., Lee, M.J., Hong, J., Hao, X., Yang,
C.S. (2008). Effect of genistein on the bioavailability and intestinal cancer
chemoprevention activity of (-)-epigallocatechi-3-gallate. Carcinogenesis 29,
2019-2024.
Lee, M.J., Maliakal, P., Chen, L., Meng, X., Bondoc, F.Y., Prabhu, S., Lambert,
G., Mohr, S., Yang, C.S. (2002). Pharmacokinetics of tea catechins after
ingestion of green tea and (-)-epigallocatechin-3-gallate by humans:
formation of different metabolites and variability. Cancer Epidemiology
Biomarkers & Prevention 11, 1025-1032.
Li, H., Li, F., Yang, F., Fang, Y., Xin, Z., Zhao, L., Hu, Q. (2008). Size effect of
Se-enriched green tea particles on in vitro antioxidant and antitumor activities.
J Agric Food Chem. 56, 4529-4533.
Maeda-Yamamoto, M., Kawahara, H., Matsuda, N., Nesumi, K., Sano, M., Tsuji,
K., Kawakami, Y., Kawakami, T. (1998). Effects of tea infusions of various
varieties or different manufacturing types on inhibition of mouse mast cell
activation. Biosci Biotechnol Biochem. 62, 2277-2279.
Maeda-Yamamoto, M., Sano, M., Matsuda, N., Miyase, T., Kawamoto, K.,
Suzuki, N., Yoshimura, M., Tachibana, H., Hakamata, K. (2001). The change
of epigallocatechin-3-O- (3-O-methyl) gallate contents in tea of different
varieties, tea seasons of crop and processing method. Nippon Shokuhin
Kagaku Kougaku Kaishi (Japanese) 48, 64-68.
Maeda-Yamamoto, M., Nagai, H., Asai, K., Moriwaki, S., Horie, H., Kohata, K.,
Tachibana, H., Miyase, T., Sano, M.(2004). Changes in
Epigallocatehin-3-O-(3-O-methyl) gallate and strictinin contents of tea
(Camellia sinensis L.) cultivar ‘Benifuuki in various degree of maturity and
leaf order. Food Science and Technology Research 10, 186-190.
Maeda-Yamamoto, M., Inagaki, N., Kitaura, J., Chikumoto, T., Kawahara, H.,
Kawakami, Y., Sano, M., Miyase, T., Tachibana, H., Nagai, H., Kawakami, T.
(2004). O-methylated catechins from tea leaves, inhibit multiple protein
kinases in mast cells. J Immunology 172, 4486-4492.
Maeda-Yamamoto, M., Ema, K., Shibuichi, I. (2007). In vitro and in vivo
anti-allergic effects of ‘Benifuuki’ green tea containing O-methylated catechin
and ginger extract enhancement. Cytotechnology 55,135-142.
Maeda-Yamamoto, M., Ema, K., Tokuda, Y., Monobe, M., Tachibana, H.,
Sameshima, Y., Kuriyama, S. (2011). Effect of green tea powder (Camellia
sinensis L. cv. Benifuuki) particle size on O-methylated EGCG absorption in
rats; The Kakegawa Study. Cytotechnology 63,171-179.
Matsuo, N., Yamada, K., Shoji, K., Mori, M., Sugano, M. (1997). Effect of tea
polyphenols on histamine release from rat basophilic leukemia (RBL-2H3)
cells; structure-inhibitory activity relationship. Allergy 55, 58-64.
O'Brien, N., Cummins, E. (2010). Ranking initial environmental and human
health risk resulting from environmentally relevant nanomaterials. J Environ
Sci Health A Tox Hazard Subst Environ Eng. 45, 992-1007.
Sano, M., Suzuki, M., Miyase, T., Yoshino, K., Maeda-Yamamoto, M. (1999).
Novel antiallergic catechin derivatives isolated from oolong tea. J Agric Food
Chem. 47, 1906-1910
Sawamura, S., Haraguchi, Y., Ikeda, H., Sonoda, J. (2010). Properties and
shapes of Matcha with various milling method. Nippon Shokuhin Kagaku
Kogoku Kaishi (Japanese) 57, 304-309.
Suzuki, M., Yoshino, K., Maeda-Yamamoto, M., Miyase, T., Sano, M. (2000)
Inhibitory Effects of Tea Catechins and O-methylated Derivatives of
(-)-Epigallocatechin-3-O-gallate on Mouse Type-IV Allergy. J Agric Food
Chem. 48, 5649-5653.
Suzuki, T., Yamazaki, N., Sada, Y., Oguni, I., Moriyasu, Y. (2003). Tissue
distribution and intracellular localization of catechin in tea leaves. Biosci.
Biotechnol. Biochem. 67, 2683-2868.
Tachibana, H., Kubo, T., Miyase, T., Tanino, S., Yoshimoto, M., Sano, M.,
Maeda-Yamamoto, M., Yamada, K. (2001). Identification of an Inhibitor for
interleukin 4-induced e germline transcription and antigen-specific IgE
production in vivo. Biochem Biophys Res Commun. 280, 53-60.
Yamashita, K., Suzuki, Y., Matsui, T., Yoshimura, T., Yamaki, M.,
Suzuki-Karasaki, M., Hayakawa, S., Shimizu, K. (2000). Epigallocatechin
gallate inhibits histamine release from rat basophilic leukemia (RBL-2H3)
cells; role of tyrosine phophorylation pathway. Biochem Biophysic Res
Commun. 274, 603-608.
Yasue, M., Ikeda, M., Nagai, H., Sato, K., Mitsuda, H., Maeda-Yamamoto, M.,
Yabune, M., Nakagawa, S., Kajimoto, Y., Kajimoto, O.(2005). The clinical
effects and safety of the intakes of Benifuukigreen tea in patients with
perennial allergic rhinitis. Nihon Rinsho Eiyougakkai Zasshi (Japanese) 27,
33-51.
Figure legends
Figure 1. Plasma unconjugated EGCG and EGCG3”Me concentration versus
time profiles after oral administration of Benifuuki green tea beverage. Each
point represents the average of six subjects, and the cross-vertical bars
represent SD of the mean. All subjects drank Benifuuki green tea containing
43.5mg of EGCG and 8.5mg of EGCG3”Me within 3 minutes.
Figure 2. The effects of Benifuuki green tea and additive ginger extract on the
symptom score of seasonal allergic rhinitis. All subjects drank 1.5g of each tea
powder, Benifuuki green tea (closed circle) , Benifuuki green tea containing
30mg of ginger extract (open triangle), and Yabukita green tea (open square),
with water twice a day for 13 weeks. Christcross represents cedar pollen
number. Each point represents the average of nine subjects every two weeks
and the cross-vertical bars represent SD of the mean.
(A) blowing nose (0 (0 time)-4 (more than 21 times)), (B) itching eyes (0
(no)-4 (severe)), (C) nasal symptom medication score (SMS).
*,** Significantly different from the placebo group (*P<0.05, ** P<0.01).
Figure 3. Absorption of catechins versus time profiles after the oral
administration of 4 types of Benifuuki green tea in rats i.g..
BGP 2.86 µm: Benifuuki green tea powder with a mean particle size of 2.86
µm, BGP 18.6 µm: Benifuuki green tea powder 18.6 µm, BGP 76.1 µm:
Benifuuki green tea powder 76.1 µm, BGI: Benifuuki green tea infusion.
Absorption was calculated as follows;
Absorption = ( plasma concentration (µg/L) /1000 * blood volume (rat
weight(g) * 70 / 1000)) /(intake (mg) *1000)*100 )
Each point represents the mean of five rats, and the cross-vertical bars
represent the SD of the mean. All rats were given Benifuuki green tea
within 1 minute via gavage.
(A): EGCG3Me, (B): EGCG, (C): ECG, (D): EGC, (E): EC
Figure 4. The change in the green tea powder particle size by crushing time
using ball mill (Kobayashi, 2008).
C: control powder grounded by stone-mill
Ball-mill and stone-mill were produced by Ikeda seichakikaiten Co., Ltd.
Green tea used was crude tea manufactured by Super High Moisture
Atmosphere (SHIMA) manufacturing procedure.
Titles of Tables
Table 1. Pharmacokinetic parameters of plasma catechins after i.g.
administration of Benifuuki green tea in rats
Table 2. AUC (the area plasma concentration-time curve) for catechins after oral
administration of various types of Benifuuki green tea in humans
Table 3. Absorption for catechins after 1hr oral administration of various types
of Benifuuki green tea in humans
Fig.1
Fig.2
Fig.3
Fig.4
... The nutrients in matcha tea are 60-70% insoluble ingredients such as fat-soluble vitamins, insoluble dietary fibers, chlorophylls, and proteins. While the soluble ingredients constitute 30-40% which includes polyphenols, water-soluble vitamins, caffeine, water-soluble dietary fibers, amino acids, saponin, and minerals (Maeda-Yamamoto et al., 2013). Given their unique farming and harvesting processes, the concentrations of bioactive compounds are higher in matcha tea than in other types of green tea. ...
Article
Full-text available
Matcha is a powdered form of Japanese green tea that has been gaining global popularity recently. Matcha tea has various health benefits, including an enhancing effect on cognitive function, cardio-metabolic health, and anti-tumorogenesis. To date, randomized clinical trials (RCT) showed that matcha decreases stress, slightly enhances attention and memory, and has no effect on mood. Results regarding the effect of matcha on cognitive function are contradictory and more RCTs are warranted. The cardio-metabolic effects of matcha have only been studied in animals, but findings were more homogenous. Consuming matcha with a high-fat diet resulted in decreased weight gain velocity, food intake, improved serum glucose and lipid profile, reduced inflammatory cytokines and ameliorated oxidative stress. Evidence regarding the anti-tumor function of matcha is very limited. Findings showed that matcha can affect proliferation, viability, antioxidant response, and cell cycle regulation of breast cancer cells. Nonetheless, more studies are needed to examine this effect on different types of cancer cells, and there is also a need to verify it using animal models. Overall, the evidence regarding the effect of matcha tea on cognitive function, cardio-metabolic function, and anti-tumor role is still limited, and conclusions cannot be drawn.
Article
Full-text available
Scope: Green tea is associated with decreased risk for cardiovascular disease and stroke. Matcha is a special kind of powdered green tea known for its use in the Japanese tea ceremony. Due to its influence on lipoprotein parameters, it has been postulated to exert antiatherogenic effects. We investigated whether it modulates the HDL function and thereby influences the atherogenic process in an animal model with a strong influence on humans' situation. Methods and results: After a pretreatment phase based on a standard diet, ten female NZW rabbits were fed a high-fat diet for 20 weeks. The treatment group was additionally administered 1% matcha during the whole experiment. Long-term matcha treatment led to lowered HDL cholesterol, impaired cholesterol transport manifested by reduced in vitro cholesterol efflux capacity, reduced CETP-mediated cholesterol ester (CE) transfer between HDL and triglyceride-rich particles, and reduced macrophage-specific in vivo transfer, where we observed increased absorption of cholesterol in the liver but a decreased secretion into bile. Pulse wave velocity, assessed by nuclear magnetic resonance, was increased in matcha-treated animals, and a similar trend was observed for atherosclerotic lesion formation. Conclusion: Long-term matcha green tea treatment of hypercholesterolemic rabbits caused impaired reverse cholesterol transport and increased vascular stiffness, and susceptibility for atherosclerotic lesion development. This article is protected by copyright. All rights reserved.
Article
Full-text available
The content of epigallocatechin-3-O-(3-O-mehtyl) gallate (EGCG3″Me), which has been observed an anti-allergic action in vitro and in vivo (mouse) and present in tea leaves, was determined in different tea varieties, at different cropping seasons and upon various processing methods. Benihomare and two of its offsprings, Benifuuki and Benifuji contained as much as 1% of EGCG3″Me in their green teas and slightly less in their semi-fermented teas. Surprisingly, EGCG3″Me was absent in Izumi, a third offspring of Benihomare. A higher concentration of EGCG3″Me was found on or after the second crop in Benihomare and Benifugi. Furthermore, EGCG3″Me was not detected in black tea (fermented tea) manufactured from Benihomare and Benifugi. Thus, to benefit the anti-allergic effect of EGCG3″ Me, green or semi-fermented teas, made from Benihomare, Benifuuki and Benifugi at second crop onwards, should be consumed.
Article
Full-text available
We previously found that the O-methylated derivative of (-) epigallocatechin 3 O-gallate (EGCg), (-)-epigallocatechin 3 O-(3-O-methyl)-gallate (EGCG"3Me) has potent antiallergic activity The high-affinity IgE receptor, FcepsilonRI, is found at high levels on basophils and mast cells and plays a key role in a series of acute and chronic human allergic reactions To understand the mechanism of action for the antiallergic EGCG"3Me, the effect of EGCG"3Me on the cell surface expression of FcepsilonRI in human basophilic KU812 cells was examined Flow cytometric analysis showed that EGCG"3Me was able to decrease the cell surface expression of FcepsilonRI Moreover, immunoblot analysis revealed that total cellular expression of the FcepsilonRI alpha chain decreased upon treatment with EGCG"3Me FcepsilonRI is a tetrameric structure comprising one alpha chain, one beta chain, and two gamma chains The level of mRNA production of each subunit in KU812 cells was investigated EGCG"3Me reduced FcepsilonRI alpha and gamma mRNA levels The cross-linkage of FcepsilonRI causes the activation of basophils which leads to the secretion of inflammatory mediators including histamine EGCG"3Me treatment inhibited the FcepsilonRI cross-linking induced histamine release These results suggested that EGCG"3Me can negatively regulate basophil activation through the suppression of FcepsilonRI expression.
Article
Full-text available
We have investigated the particle size, degree of circularity, extensibility, flowability and foaming properties of matcha in terms of the milling method. Matcha samples were ground by a stone-mill, a ball-mill and a jet-mill, and the matcha ground by a ball-mill was classified by 10μm diameter into fine and coarse matcha with an airflow-type classifier. The median diameter of the matcha ground by a stone-mill and a ball-mill and a fine matcha was 15-20μm, and that of a matcha ground by a jet-mill and a fine matcha was under 5μm. The degree of circularity of a matcha ground by a jet-mill was higher than that by other milling machines. A fine matcha and the matcha ground by a jet-mill showed a higher extensibility compared to others. Matcha ground by a jet-mill showed good flowability compared to a fine one. A fine matcha showed a good foaming property compared to a coarse one. Therefore, we expect the improvement of utility since matcha ground by a jet-mill showed a predominant flowability, extensibility and foaming property because of its small diameter and high degree of circularity.
Article
(-)-Epigallocatechin-3-gallate (EGCG), from green tea (Camellia sinensis), has demonstrated chemopreventive activity in animal models of carcinogenesis. Previously, we reported the bioavailability of EGCG in rats (1.6%) and mice (26.5%). Here, we report that cotreatment with a second dietary component, piperine (from black pepper), enhanced the bioavailability of EGCG in mice. Intragastric coadministration of 163.8 mumol/kg EGCG and 70.2 mumol/kg piperine to male CF-1 mice increased the plasma C,a and area under the curve (AUC) by 1.3-fold compared to mice treated with EGCG only. Piperine appeared to increase EGCG bioavailability by inhibiting glucuronidation and gastrointestinal transit. Piperine (100 mumol/L) inhibited EGCG glucuronidation in mouse small intestine (by 40%) but not in hepatic microsomes. Piperine (20 mumol/L) also inhibited production of EGCG-3"-glucuronide in human HT-29 colon adenocarcinoma cells. Small intestinal EGCG levels in CF-1 mice following treatment with EGCG alone had a C-max = 37.50 +/- 22.50 nmol/g at 60 min that then decreased to 5.14 +/- 1.65 nmol/g at 90 min; however, cotreatment with piperine resulted in a C-max = 31.60 +/- 15.08 nmol/g at 90 min, and levels were maintained above 20 nmol/g until 180 min. This resulted in a significant increase in the small intestine EGCG AUC (4621.80 +/- 1958.72 vs. 1686.50 +/- 757.07 (nmol/g(.)min)). EGCG appearance in the colon and the feces of piperine-cotreated mice was slower than in mice treated with EGCG alone. The present study demonstrates the modulation of the EGCG bioavailablity by a second dietary component and illustrates a mechanism for interactions between dietary chemicals.
Article
Epigallocatechin-3-gallate (EGCG), the most abundant catechin in green tea (Camellia sinensis), has shown cancer preventive activity in animal models. The bioavailability of EGCG in the most commonly used animal species, mice, is poorly understood. Moreover, the pharmacokinetic parameters of EGCG have not been reported previously in mice. Here we report that after administration of EGCG intravenously at 21.8 mumol/kg or intragastrically at 163.8 mumol/kg, the peak plasma levels of EGCG in male CF-1 mice were 2.7 +/- 0.7 and 0.28 +/- 0.08 mumol/L, respectively. EGCG was present mainly (50-90%) as the glucuronide. The plasma bioavailability of EGCG after intragastric administration was higher than previously reported in rats (26.5 +/- 7.5% vs. 1.6 +/- 0.6%). The conjugated EGCG displayed a shorter t(1/2) (82.8-211.5 vs 804.9-1102.3 min) than unconjugated EGCG (P < 0.01, Student's t test). EGCG was present in the unconjugated form in the lung, prostate and other tissues at levels of 0.31-3.56 nmol/g after intravenous administration. Although intragastric administration resulted in lower levels in most tissues compared with intravenous administration (e.g., 0.006 +/- 0.004 vs. 2.66 +/- 1.0 nmol/g in the lung), the levels in the small intestine and colon were high at 45.2 +/- 13.5 and 7.86 +/- 2.4 nmol/g, respectively. This is the first report of the pharmacokinetic parameters of EGCG in mice. Such information provides a basis for understanding the bioavailability of EGCG in mice and should aid in understanding the cancer preventive activity of EGCG. J. Nutr. 133: 4172-4177, 2003.
Article
The production of Matcha for processed food is increasing in recent years. However, Matcha has many problems for use in processed food, such as bad suspension, color deterioration, and high costs. We developed fine Matcha to overcome these problems, and investigated its properties. Fine Matcha was produced by two methods : classification of normal Matcha, and crushing by jet-mill. The particle size of normal Matcha ranges widely from 1 μm to 100 μm, but that of fine Matcha has a narrow range of from 1 μm to 20 μm. The particle diameter of fine Matcha is 3.2 μm, and that of normal Matcha is 11.5 μm. Fine Matcha suspended in water has a smaller sedimentation rate than normal Matcha. In addition, fine Matcha used in buckwheat noodles (soba noodles) improved the colorimetry of the noodles in comparison with normal ones. Finally, fine Matcha added to dough was found to cause the bread to rise better than normal. Thus, fine Matcha is useful for processed food in producing less sedimentation, improving the colorimetry, and augmenting the rising of bread dough.
Article
We have reported that O-methylated EGCGs (epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3″Me) and epigallocatechin-3-O-(4-O-methyl) gallate) and strictinin had anti-allergic action, and that the tea (Camellia sinensis L.) cultivar 'Benifuki' was rich in EGCG3″Me. However, changes in the content of anti-allergic components in tea leaves during growth are unclear. Therefore, here we determined the contents of EGCG3″Me and strictinin at various points of maturity and in the leaf order of 'Benifuki' tea in the first and second crop seasons. The EGCG3″Me content was high and caffeine content decreased in fully-matured tea leaves late in the first crop season or under the third leaf during the second crop season. The amount of EGCG3″Me contained in the stem was low. A higher concentration of strictinin was found in younger new shoots in the first crop, and the content greatly decreased during the second crop season. Thus, to obtain the anti-allergic benefits of EGCG3″Me, green tea should be made from fully-matured 'Benifuki' leaves under the fourth shoot late in the first crop or under the third shoot in the second crop, refined by removing stems. This green tea is lower in caffeine as compared with that made from younger shoots. On the contrary, to utilize strictinin, green tea should be made using younger new shoots of 'Benifuki.'.