Field of Invention:
Present invention relates to a herbal composition for therapeutic support to digestive system,
cardiac system, and nutritive for nervous system, mental health along with detoxification of
body and anti-oxidant effect.
Background of Invention:
Herbal based compositions have been conventionally used as drug, nutrients and drinks. The
selected species of herbs are widely distributed throughout the temperature zones of Asia and
Europe and throughout cold aid zones of high altitudes of Asia. In India, these herbs are
distributed from plains to high altitudes of Himalayas. In "Amchi" system of medicine
(Tibetan system of medicine) for treatment of various ailments viz. fever, cough and cold,
rheumatism, asthma, health tonic, stomach complaints etc. Flowers, fruits/seeds, leaves and
root/rhizomes of these herbs are used for formulation of medicated herbal tea. The herbal tea
contains vitamins. Flavonoids and phenols etc. which act as anti-oxidant agents improve
body strength and restores memory and improve vital Health. The natural herbs are source of
various bioactive-compounds, edible colors and taste which are not easy to consume directly.
According to the same patents on herbal tea this process for formulation and composition are
quite different. All those selected herbs used in formulation of this particular herbal tea are
wild or wild relatives of plants used by human beings for various uses.
However, no patent is reported so far on processing of formulation of medicated herbal tea in
synergic from selected plants so as to prepare a medicated herbal tea having potential
antioxidants, with goods taste, natural color suitable for direct consumption by human beings,
without adding any artificial color, flavor or stabilizing agents.
Brief description of Drawings:
Figure 1 shows table showing individual body weight in different groups up to 7 days after
administration of HTCP at 200 and 400 mg dose.
3
Figure 2 shows table showing status of Mean body weight from initial day to 7th day in
various groups. All data are mean ± SD of six animals in each group. Group I is carrageenan
induced paw edema group whereas group II is standard drug ibuprofen treated group (400mg/
kg) +carrageenan induced paw edema. Group III and Group IV are polyherbal tea treated
groups at dose levels (200mg and 400mg/kg) +carrageenan induced paw edema group The
statistical data showed the significant increased between initial day to 7th day in each group.
Where * is significant (P<0.01) and *** is highly significant (P<0.0001).
Figure 3 shows body weight measurement administration of Carrageenan and HTCP drug
treatments. All data are Mean ± SD of six animals each group. Group I is carrageenan
induced paw edema group whereas group II is ibuprofen treated group (standard drug) and
group III and IV are herbal tea coarse powder treated group at dose levels of 200 and 400
mg/kg.
Figure 4 shows a table showing status of mortality and clinical sign of toxicity upto 7 days
after HTCP administration at dose level 200 and 400 mg/kg.
Figure 5 shows a table for Paw Edema measurement in each group of animals after
administration of carrageenan and test item HTCP at dose level 200 and 400 mg/kg.
Figure 6 shows status of anti-inflammatory effect in the term of paw edema by carrageenan
in all groups.
Figure 7 shows a table showing Individual body weight in different groups up to 10days after
administration of HTCP at 200 and 400 mg dose levels.
Figure 8 shows a table showing status of Mean body weight from initial day and 5th and 10th
day in each group. All data are mean SD of six animals in each group. Group I is Normal
saline treated group whereas group II is Scopolamine induced group (0.7mg/ kg) and Group
III and Group IV are scopolamine (0.7mg/kg) plus poly- herbal tea treated groups at dose
levels (200mg and 400mg/kg). The statistical data showed the significant increased between
4
initial day vs 5th, 10th day and 5th vs 10th day in each group. Where * is significant (P<0.05)
and ** is significant (P<0.001); *** is highly significant (P<0.0001).
Figure 9 shows status of Body weight in scopolamine induced and HTCP treated groups at
dose levels (200 and 400mg/kg).
Figure 10 shows Status of mortality and clinical sign of toxicity upto 7 days after HTCP
administration at dose level 200 &400 mg/kg. No any mortality and clinical sign of toxicity
found in each group of animals after single administration of poly-herbal tea HTCP at dose
levels 200 &400 mg/kg up to 7 days. Scopolamine induced group (II) show the symptom of
liturgy. Where 0 is no mortality.
Figure 11A shows status of feed and water intake in each group.
Figure 11B shows status of feed and water intake in each group.
Figure 12 shows Status of AChE enzyme activity in all group.
Summary of Invention:
According to an aspect of the invention, the composition comprises of components, said
components are leaves of Ocimum sanctum, leaves of Chamillia sinesis, bark of Cinamonum
zeylanicum, leaves of Cinamonum zeylanicum, bark of Terminalia arjuna, root of Withania
somnifera, rhizome of Zingiber officinalis, seeds of Piper nigrum, flower bud of Syzygium
aromaticum., and acceptable herbal excipients.
According to an aspect of the invention, the composition comprises of herbal excipients for
aroma and enhancing flavor.
According the herbal excipients can be selected from a group consisting of seeds of Anethum
Sowa, seeds of Foeniculum vulgare, seeds of Cuminum cyminum, fruits of Elettaria
cardamonum and Floral part of Myristica fragrans.
5
According to an aspect of the invention, wherein the amount of leaves of Ocimum sanactum
is in a range of 300 mgs to 500 mgs, bark of Cinamonum zeylanicum are in a range of 50 mgs
to 300 mgs, leaves of Cinamonum zeylanicum are in a range of 50 mgs to 300 mgs root of
Withania somnifera is in a range of 100 mgs to 400 mgs, bark of Terminallia arjuna is in a
range of 100 mgs to 400 mgs, rhizome of Zingiber officinalis is in range of 50 mgs to 250
mgs, fruit of Piper nigrum is in range of 50 mgs to 200 mgs, flower bud of Syzygium
aromaticum in a range of 40 mgs to 200 mgs, and leaves of Chamillia sinesis is in a range of
300 mgs to 500 mgs per 2 gm of composition.
According to an aspect of the invention, the composition comprises seeds of Anethum sowa
in a range of 40 mgs to 200 mgs, seeds of Foeniculm vulgrae is in a range of 50 mgs to 200
mgs, seeds of Cuminum cyminum are in a range of 40 mgs to 200 mgs, fruits of Elettaria
cardamonum in a range of 40 mgs to 200 mgs, Floral part of Myritica fragrans in a range of
10 mgs to 50 mgs, per 2 gm of composition.
According to an aspect of the invention, there is provided a process for preparation for herbal
tea, said method comprising drying and grinding of leaves of Ocimum sanctum, leaves of
Chamillia sinesis, bark of Cinamonum zeylanicum, leaves of Cinamonum zeylanicum, bark of
Terminalia arjuna, root of Withania somnifera, rhizome of Zingiber officinalis, seeds of
Piper nigrum, flower bud of Syzygium aromaticum, and acceptable herbal excipients.
According to an aspect of the invention, the parts are dried and grinded to a uniform size of 2
to 3 mm.
According to an aspect of the invention, the leaves of Ocimum sanctum, leaves of Chamillia
sinesis, bark and leaves of cinnamon, bark of Terminalia arjuna, root of Withania somnifera,
dry ginger, seeds of Piper nigrum, seeds of Foeniculum vulgare, seeds of Soya, flower buds
of clove, seeds of cumin, seeds of cardamom and Floral part of Myristica fragrans are mixed
together at a temperature of from 32 degree centigrade to 38 degree centigrade.
6
According to an aspect of the invention, drying is done at a temperature of shade drying at
25-35 degree centigrade and followed by drier at 48 degree centigrade to 58 degree
centigrade.
According to an aspect of the invention, the dosage range of the composition ranges from the
dosage range of the composition ranges from 4.02 gms to 5.94 gms in three divided dose at
the minimum interval of 5-6 hrs.
Detailed description of invention
Present invention relates to a herbal tea composition having adaptogenic effect, and is a
potential anti-oxidant, provide energy to body, refreshing, anti-fatigue to fight from the
oxidative stress and to protect the body from rapid deterioration and ageing. The composition
helps to support the cardiac, brain and digestive health.
The herbal tea composition has ingredients which work in synergy in order to enhance the
anti-oxidant and nutritive effect simultaneously to enjoying health benefits.
According to an embodiment of the invention, the herbal tea composition can also be used as
an additive and as a flavoring agent for other food products.
The composition comprises of leaves of Ocimum sanctum, leaves of Chamillia sinesis, bark
of Cinamonum zeylanicum, leaves of Cinamonum zeylanicum, bark of Terminalia arjuna,
root of Withania somnifera, rhizome of Zingiber officinalis, seeds of Piper nigrum, flower
bud of Syzygium aromaticum, and acceptable herbal excipients. The herbal excipients are
herbal and botanical in nature and function and can be selected from a group consisting of
seeds of Anethum Sowa, seeds of Foeniculum vulgare, seeds of Cuminum cyminum, fruits of
Elettaria cardamonum and Floral part of Myristica fragrans.
7
The amount of leaves of Ocimum sanactum is in a range of 300 mgs to 500 mgs, bark of
Cinamonum zeylanicum are in a range of 50 mgs to 300 mgs, leaves of Cinamonum
zeylanicum are in a range of 50 mgs to 300 mgs root of Withania somnifera is in a range of
100 mgs to 400 mgs, bark of Terminallia arjuna is in a range of 100 mgs to 400 mgs,
rhizome of Zingiber officinalis is in range of 50 mgs to 250 mgs, fruit of Piper nigrum is in
range of 50 mgs to 200 mgs, leaves of Chamillia sinesis is in a range of 300 mgs to 500 mgs,
frlower bud of Syzygium aromaticum in a range of 40 mgs to 200 mgs, per 2 gm of
composition.
The herbal excipients which may function to improve the taste and aroma of the composition,
such as seeds of Anethum sowa are in a range of 40 mgs to 200 mgs, seeds of Foeniculm
vulgrae is in a range of 50 mgs to 200 mgs, seeds of Cuminum cyminum are in a range of 40
mgs to 200 mgs, fruits of Elettaria cardamonum in a range of 40 mgs to 200 mgs, Floral part
of Myritica fragrans in a range of 10 mgs to 50 mgs, per 2 gm of composition.
The process for preparation of herbal composition of present invention, comprises drying,
followed by grinding of all components of the herbal tea. Optionally herbal excipients may
be added to enhance flavor and aroma to the tea. Accordingly, tea of various flavors can be
prepared by varying the kind of herbal excipients.
According to an aspect of the invention, there is provided a process for preparation for herbal
tea, said method comprising drying and grinding of leaves of Ocimum sanctum, leaves of
green tea, bark of Cinamonum zeylanicum, leaves of Cinamonum zeylanicum, bark of
Terminalia arjuna, root of Withania somnifera, rhizome of Zingiber officinalis, seeds of
Piper nigrum, flower bud of Syzygium aromaticum, and acceptable herbal excipients.
According to an aspect of the invention, desired plant parts of all the components and
optionally alongwith excipients are dried and grinded to a uniform size of 2 to 3 mm and then
mixed together at room temperature preferably ranging from 32 degree centigrade to 38
degree centigrade.
8
According to an aspect of the invention, the dosage range of the composition ranges from the
dosage range of the composition ranges from 4.02 gms to 5.94 gms in three divided dose at
the minimum interval of 5-6 hrs.
According to an aspect of the invention, drying is done at a temperature of shade drying at
25-35 degree centigrade and followed by drier at 48 degree centigrade to 58 degree
centigrade.
Experiment:
A composition is prepared by comprising the following components:
Leaves of Ocimum sanactum 400 mg,
Leaves of Chamillia sinesis 400 mg,
Root of Withania somnifera 200 mg,
Bark of Terminallia arjuna 200 mg;
Bark of Cinamonum zeylanicum 100 mg
Leaves of Cinamonum zeylanicum 100 mg
Rhizome of Zingiber officinalis 100 mg;
Fruit of Piper nigrum 100 mg,
Seeds of Foeniculm vulgrae 100 mg;
Seeds of Cuminum cyminum 60 mgs;
Seeds of Anthem Sowa 80 mg,
Flower bud of Syzygium aromaticum 60 mgs;
Fruit of Elettaria cardamonum 80 mgs;
Floral part of Myritica fragrans 20 mgs.
Experiment 1:
Evaluating anti-inflammatory effect of poly-herbal tea herbal tea coarse powder (HTCP) in
wistar rat model. Total 24 male animals were selected and acclimatized for 5 to 7 days and
9
animals were randomized into four groups of six animals each. The acute inflammation was
induced in the right hind paw of all animals by administration of carrageenan 0.1 ml (1% w/v
suspension of carrageenan) i.p into the sub-plantar surface of hind limb paw and
inflammation was measured in all groups by digital vernier calliper. Group I was received
only carrageenan and act as negative control where as group II to group IV received standard
drug ibuprofen (400mg/kg) and 200 and 400 mg/kg dose levels of poly-herbal tea Herbal Tea
Coarse Powder (HTCP) before one hour administration of carrageenan in right paw of
animals.
Result:
findings showed that the body weight of each groups were found significantly increased
(P<0.01; P<0.001) manners from initial day of each group. Group I animals increased the
paw edema at 30-240 minutes intervals whereas group II to IV animals showed the decreased
the paw edema in all treated groups in comparison to group I animals whose received only
carrageenan into the subplantar surface of hind limb paw. When compared the both dose
levels of HTCP oral treated group (III and IV) with standard drug ibuprofen treated group at
the different time intervals, the paw edema was reduced in time dependent manners and
almost equal to standard drug manners. When both poly-herbal tea dose levels (200 and 400
mg/kg) were compared to each other in time dependent manners, the paw edema was found
highly inhibit in 200 mg/kg poly-herbal tea (HTCP) and it is almost equal to standard drug
ibuprofen.
MATERIALS AND METHODS
Test item Information (Vehicle control)
Name of test item : CM Cellulose (vehicle control)
Description : White to off white powder
Batch No.: C153934
Sample quantity 2.0g
Manufacturing Date 27-08-2016
Expiry Date 26-08-2018
10
Test item information (Carragennan)
Name of test item : Carrageenan
Description : White to off white powder form
Lot No.: 257633
Sample quantity 100 g
Manufacturing Date February 2016
Expiry Date February 2018
Test item information (ibuprofen)
Name of test item : Ibuprofen
Description : Dark Pink Tablet Form
Batch No.: B.No. 9369
Sample quantity 400 mg
Manufacturing Date October 2016
Expiry Date September 2019
Test item information (Herbal Tea Coarse Powder)
Description : Herbal Tea Coarse Powder
pH 5.14
Batch No.: VHT-133
Solubility in water: 68.21%
Solubility in solvent 43.32%
Density Before compaction : 0.3678
Density After Compaction : 0.5231
Manufacturing Date 30-07-2016
Expiry Date Two years from date of manufacturing
Experiments were performed on specie Rattus norvegicus of Albino rats.
Test item/ material herbal tea coarse powder (HTCP) was powder form so, it was dissolved
into 1% CM cellulose. Whereas other test item Carrageenan and standard drug ibuprofen was
dissolved into distilled water.
The anti-inflammatory activity was performed by the protocol of carrageenan induced paw
edema in animal model. In this study, total 24 male wister rats (weighting 120 to 150 g) was
selected. All animals were acclimatized. Standard animal house environment for at least 5 to
7 days before initiation of experiment. After the acclimatization, animals were randomized
11
on the basis of body weight into four groups of 6 animals each. All animals were located into
following groups
Groups No of animals/
Sex
Dose regiman Dose(mg/kg
body weight)
Test Item
I 6/M Single 0.1ml Carragenam
II 6/M Single 400 mg/kg Ibuprofen
III 6/M Single 200 mg/kg HTPC
IV 6/M Single 400 mg/kg HTPC
Group I was received carrageenan 0.1ml (1%w/v) whereas group II, III and IV were received
test item ibuprofen (400mg/kg) and herbal tea coarse powder (200 and 400 mg/kg) at dose
levels before one hour administration of carrageenan. The acute inflammation was induced in
the right hind paw of all animals by administration of carrageenan 0.1 ml (1% w/v
suspension of carrageenan) i.p into the sub-plantar surface of hind limb paw. The diameter of
the injected paw was measured using digital vernier calliper at time 30, 60, 120, 180 and 240
minutes. The paw edema as acute inflammation was characterized by increase in the diameter
of the injected paw. Each group of animals were recorded body weight and monitored
mortality, clinical sign of toxicity up to seven days after oral administration of test item
herbal Tea Coarse Powder (HTCP).
Group I was received carrageenan 0.1ml (1%w/v) whereas group II, III and IV were received
test item ibuprofen (400mg/kg) and herbal tea coarse powder (200 and 400 mg/kg) at dose
levels before one hour administration of carrageenan. The acute inflammation was induced in
the right hind paw of all animals by administration of carrageenan 0.1 ml (1% w/v
suspension of carrageenan) i.p into the sub-plantar surface of hind limb paw. The diameter of
the injected paw was measured using digital vernier calliper at time 30, 60, 120, 180 and 240
minutes. The paw edema as acute inflammation was characterized by increase in the diameter
of the injected paw. Each group of animals were recorded body weight and monitored
mortality, clinical sign of toxicity up to seven days after oral administration of test item
herbal Tea Coarse Powder (HTCP). administration/ of test items. When the data was
12
compared from initial day to 7th days, the body weight was found statistically (P<0.01,
P<0.0001) significant increase in manner.
The data are presented in table shown in Figure 1, Figure 2 and graph shown in Figure 3.
No any mortality were found in all groups of animals after administration of test item for
seven days. The resulting data are presented in Figure 4.
No any clinical sign of toxicity was observed except paw edema after administration of test
item carrageenan. All animals were found normal up to seven days. The results are presented
in table shown in Figure 4.
Carrageenan induced Paw edema measurement (Anti-inflammatory effect)
In the present study, the sub-plantar injection of 1% carrageenan suspension produced a local
edema statistically (p<0.0005) significant increased with its peak at 3h in the group I animals
(carrageenan induced group) in comparison to initial time 30 minutes and time dependent
manners. The paw edema was slightly reduced at 4hours (240 minutes) in carrageenan
induced group. The paw edema of standard drug ibuprofen treated group (group II) was
significantly (p<0.001) reduced after 4 hours (240minutes) in comparison to initial time (30
minutes). The % inhibition of paw edema was found 26.5% in ibuprofen treated group after
240minutes interval as compared with 30 minutes interval. When the % inhibition of paw
edema was compared between carrageenan treated group and ibuprofen treated group at 4
hours intervals, the % inhibition of paw edema was found reduced 55.35% in ibuprofen
treated group. The table and graph are presented in Figure 5 and Figure 6.
The dose levels (200 and 400 mg/kg) of polyherbal tea HTCP (Herbal Tea Coarse Powder)
showed the significant (p<0.0001, 37.35%; p<0.0001, 37.20%) inhibition of carrageenan
induced paw edema over a span of 4h dose in the dependently manners. When the both dose
levels (200 and 400 mg/kg) of HTCP were compared to each other at 4 hours intervals, the
percentage inhibition of carrageenan paw edema was found 17.5% in HTCP treated group
(200 mg/kg ) in comparison to 400 mg /kg HTCP treated group (group IV). When the both
dose levels of HTCP treated groups were compared with standard ibuprofen treated group
13
(group II) at 4 hours intervals, the percentage inhibition of paw edema was found low 9.5%
in group III treated group (200 mg/kg) where as a slightly increased percentage of paw
edema 10.7% in group IV treated group (400 mg/kg). There were found statistically
significant (p<0.0001) reduced the percentage of paw edema in both dose levels of poly
herbal tea (200 and 400 mg/kg) treated groups (III and IV) when compared with carrageenan
induced group (I) at different time intervals (30 to 240minutes).
On the basis above finding it is clear indicating that the herbal tea coarse powder show antiinflammatory
effect at dose level 200 mg/kg which is almost show the effect standard drug
Ibuprofen.
Experiment 2:
Evaluating Neuroprotective activity of tea herbal tea coarse powder (HTCP) in wistar rat
model. Total 24 male animals were selected and acclimatized for 5 to 7 days and animals
were randomized into four groups of six animals each. The Alzheimer type dementia was
induced in all groups except control group by intraperitoneal administration of scopolamine
drug at dose level (0.7mg/kg) and evaluate the effect of test item Herbal Tea Coarse Powder
(HTCP) at dose level 200 and 400 mg/kg respectively in group III and group IV. The result
of finding shows that there were significant (P<0.0001) reduced the body weight in
scopolamine induced group as compared with control group. After treatment with test item
HTCP at dose level 200 and 400 mg /kg for 10 days, the body weight was found significant (
P<0.0001) increased in both group (group III and IV) and come back near to control group
when compared with scopolamine induced group (group II) . The food and water intake was
found decreased in manner up to 10 days in scopolamine induced group. After treatment with
test Item HTCP at respective dose levels, these parameters was found normal and come back
to control group. There were no mortality found in all groups along with no any clinical sign
of toxicity was found all treated group except lethargy symptom which was found in
scopolamine induced group. There was significantly (P<0.0001) enhanced acetyl
cholinestrase enzyme (AChE) in scopolamine induced group as compared with control
group. After treatment with test item HTCP at dose levels for 10 days, the enzyme activity
14
was found significantly (P<0.0001) reduced in both treated group. When compared between
both treated group for 10 days, the enzyme activity was found significant (P<0.001)
decreased in test item HTCP treated group at dose level 200 mg/kg (group III). So on the
basis of above findings, the study concluded that HTCP test item shows neuroprotective
activity at both dose levels and on
comparative study that it was found that the test item HTCP at dose level 200 mg/kg shows
better response than 400 mg/kg.
Many naturally occurring compounds have been proposed as potential therapies to slow or
prevent the progression of AD mostly by acting as antioxidants. Various studies have been
suggested that the positive effects of dietary antioxidants as an aid in potentially reducing
somatic cell and neuronal damage by free radicals. The beneficial health effects of plantderived
products have been largely attributed to polyphenolic compounds, as well as
vitamins, minerals and dietary fibers. A purpose of this study was to evaluate the
neuroprotective activity of poly-herbal tea HTCP in scopolamine induced neuronal
dysfunction wistar rat model.
Test item Information (Vehicle control)
Name of test item : CM Cellulose (vehicle control)
Description : White to off white powder
Batch No.: C153934
Sample quantity 2.0g
Manufacturing Date 27-08-2016
Expiry Date 26-08-2018
Test item Information (Scopolamine)
Name of test item : Scopolamine
Description : White clear solution in amber colour glass bottle
Lot No.: 63323-268-01
Sample quantity 0.4 mg/ml
Manufacturing Date August 2016
Expiry Date August 2018
15
Test item Information (Herbal Tea Coarse Powder)
Description : Green /Black and brown colour crushed leave powder
pH 5.14
Batch No.: VHT-133
Solubility in water: 68.21%
Solubility in solvent 43.32%
Density Before compaction : 0.3678
Density After Compaction : 0.5231
Manufacturing Date 30-07-2016
Expiry Date Two years from date of manufacturing
Test item/ material herbal tea coarse powder (HTCP) was in a powder form so, it was
dissolved into 1% CM cellulose. Whereas other test item scopolamine is liquid form so dose
was given according to individual animal body weight.
Calculated amount of test items scopolamine (0.7 mg) and HTCP( herbal tea coarse powder
200 and 400 mg/kg) were used according to individual body weight of animal administered
via oral route at dose level 10ml/kg.
In this study, total 24 male wistar rats (weighting 220 to 250 g) was selected. All animals
were acclimatized in Standard animal house environment for at least 5 to 7 days before
initiation of experiment. After the acclimatization, animals were randomized into four
groups of 6 animals each.
Groups No of
animals/ Sex
Dose
regiman
Dose(mg/kg body weight) Test Item
I 6/M Daily 10ml/Kg Normal Saline
II 6/M Daily Scopolamine (0.7 mg/kg) Scopolamine
III 6/M Daily S(0.7mg/kg)+HTCP 200mg/kg HTPC
IV 6/M Daily S(0.7mg/kg)+HTCP400mg/kg HTPC
Where S is Scopolamine
Group I received normal saline whereas group II received only scopolamine at dose level
(0.7mg/kg), whereas groups III and IV received test item scopolamine plus herbal tea coarse
16
powder (200 and 400 mg/kg) at dose levels for 10 days. Each group of animals were recorded
body weight, food and water intake along with monitored mortality, clinical sign of toxicity
daily up to 10 days after of scopolamine and as well as administration of test item herbal Tea
Coarse Powder (HTCP). At the end of experiment, all groups of animals were anaesthetized
and scarified for collection of brain organ for measurement of acetyl choline esterase enzyme
(AChE) activity.
PREPARATION OF BRAIN HOMOGENATE
After sacrificing the animals of each group by decapitation, brain was removed and rinsed
three times with ice cold isotonic saline solution. 500 mg of Brain tissue sample was
homogenized with 10 ml of ice cold 0.1M phosphate buffer (pH 7.4) and centrifuged at
10,000x g for 15min, supernatant was taken separately in Eppendorf tubes for estimation of
AChE enzyme activity
Body weight of individual groups of animals were recorded for 10 days in raw data sheet.
Feed and water intake of each groups of animals were recorded for 10 days in raw data
Sheet.
Mortality was also observed and recorded into raw data sheet for 10 days.
Clinical sign of toxicity was also observed and recorded into raw data sheet for 10days.
ESTIMATION OF ACETYLCHOLINESTERASE (AChE) ENZYME ACTIVITY
The AchE activity was assayed in brain homogenate by method of Rocha et al. The reaction
mixture consists of 50 mM phosphate buffer (pH 7.5) and 1mM of DTNB. The method is
based on the formation of the yellow anion 4,4 -dithio-bis -acid-nitro benzoic measurement at
412nm during 2 min of incubation at 25oC. The reaction was initiated by
addition of 0.8M acetylcholine. The results are expressed in micromole/mg protein.
Body Weight Measurement:
The results of present study showed that there was significant increased body in normal
17
saline treated group on 5th and 10th day from initial day. When 5th day body weight of control
group compared with 10th day, the body weight was found highly significant increase in
control group. Whereas in case of scopolamine induced group, the body weight was found
significantly (P<0.0001) decreased in scopolamine induced group.
In scopolamine plus HTCP treated groups (group III and group IV), the body weight was
found significantly increased (P< 0.001; P<0.0001) 5th and 10th day when compared from
initial day. When both treated groups were compared with scopolamine treated group
(Group II) on 5th and 10th day, the body weight was found statistically significant increased
in both HTCP treated groups at dose level 200 &400 mg/kg. The results are presented in
Figure 7, Figure 8 and Figure 9.
No mortality was found in any group of animals after intra-peritoneal administration of
scopolamine and oral administration of test item HTCP at dose levels 200 and 400mg/kg
respectively up to 10 days. In the clinical sign of toxicity parameter, the scopolamine induced
group animals shows the dullness in behavior and increased in the latency, whereas in case of
herbal tea coarse powder (HTCP) treated showed the normal behavior at dose levels 200
&400mg/kg. The resulting data are presented in Figure 10.
There was significantly decreased food and water intake status in scopolamine induced
group (group II) as compared to control group from initial day to 10th day. Initial day animals
received the 100g of feed and second day feed consumption was recorded. From 2nd day
animal received 200g of feed. The feed consumption was found almost normal in
HTCP treated group as compared to control group. The feed consumption was found
increased in the HTCP treated group (group III and IV) as compared with scopolamine
induced group. The water intake was found decreased in scopolamine induced group as
compared with control group. The HTCP treated groups of animal shows significant
increased the water intake as compared with scopolamine induced group (group II).
18
Effect of Herbal Tea Coarse Powder (HTCP) on AChE enzyme activity
There was significantly (P<0.0001) increased the AChE enzyme activity in the brain tissue
of scopolamine induced group (group II) as compared with control normal saline treated
group (group I). After treatment with test item HTCP at dose levels (200 and 400 mg/kg) for
10 days treatment, the AChE enzyme activity was found significantly (P<0.0001) decreased
in brain tissue homogenate of both treated group (group III and group IV). When the enzyme
activity was compared between both treated groups, the enzyme activity was found
significant (P<0.001) decreased in group III (HTCP at 200 mg/kg) as compared with group
IV treated group.
On the basis above finding it is clear indicating that the herbal tea coarse powder show
neuroprotective activity at dose level 200 & 400 mg/kg. On comparative analysis, the Test
Item “ HTCP ” shows better neuroprotective response at dose level 200 mg/kg than 400
mg/kg.males shows significant increased water intake as compared with scopolamine
induced group (group II). Figure 11A and 11B shows status of feed and water intake in each
groups. All data are every day consumption of feed in six animals of each group. Group I is
normal saline treated group whereas group II, group III and group IV are scopolamine and
scopolamine plus HTCP treated groups at dose levels (200 and 400mg/kg body weight).
Figure 12 shows status of AChE enzyme activity in all groups. All data are Mean SD of six
animals of each group. Group I is control group whereas group II is scopolamine induced
group (0.7mg/kg) ,group III and group IV are poly-herbal tea treated groups at dose levels
(200mg and 400mg/kg) +scopolamine induced group.

Claims:
1. A herbal composition comprises of components, said components are leaves of
Ocimum sanctum, leaves of Chamillia sinesis, bark of Cinamonum zeylanicum, leaves
of Cinamonum zeylanicum, bark of Terminalia arjuna, root of Withania somnifera,
rhizome of Zingiber officinalis, seeds of Piper nigrum, flower bud of Syzygium
aromaticum, and acceptable herbal excipients.
2. The herbal composition as claimed in claim 1, wherein said composition comprises of
herbal excipients for aroma and enhancing flavor.
3. The herbal composition as claimed in claim 1, wherein the herbal excipients can be
selected from a group consisting of seeds of Anethum Sowa, seeds of Foeniculum
vulgare, seeds of Cuminum cyminum, fruits of Elettaria cardamonum and Floral part
of Myristica fragrans.
4. The herbal composition as claimed in claim 1, wherein the amount of leaves of
Ocimum sanactum is in a range of 300 mgs to 500 mgs, bark of Cinamonum
zeylanicum are in a range of 50 mgs to 300 mgs, leaves of Cinamonum zeylanicum
are in a range of 50 mgs to 300 mgs root of Withania somnifera is in a range of 100
mgs to 400 mgs, bark of Terminallia arjuna is in a range of 100 mgs to 400 mgs,
rhizome of Zingiber officinalis is in range of 50 mgs to 250 mgs, fruit of Piper
nigrum is in range of 50 mgs to 200 mgs, flower bud of Syzygium aromaticum in a
range of 40 mgs to 200 mgs, and leaves of Chamillia sinesis is in a range of 300 mgs
to 500 mgs per 2 gm of composition.
5. The herbal composition as claimed in claim 3, wherein the composition comprises
seeds of Anethum sowa in a range of 40 mgs to 200 mgs, seeds of Foeniculm vulgrae
is in a range of 50 mgs to 200 mgs, seeds of Cuminum cyminum are in a range of 40
20
mgs to 200 mgs, fruits of Elettaria cardamonum in a range of 40 mgs to 200 mgs,
Floral parts of Myritica fragrans in a range of 10 mgs to 50 mgs, per 2 gm of
composition.
6. A process for preparation for herbal tea, said method comprising drying and grinding
of leaves of Ocimum sanctum, leaves of Chamillia sinesis, bark of Cinamonum
zeylanicum, leaves of Cinamonum zeylanicum, bark of Terminalia arjuna, root of
Withania somnifera, rhizome of Zingiber officinalis, seeds of Piper nigrum, flower
bud of Syzygium aromaticum, and acceptable herbal excipients.
7. The process as claimed in claim 6, wherein the parts are dried and grinded to a
uniform size of 2 to 3 mm.
8. The process as claimed in claim 6, wherein the leaves of Ocimum sanctum, leaves of
Chamillia sinesis, bark and leaves of cinnamon, bark of Terminalia arjuna, root of
Withania somnifera, dry ginger, seeds of Piper nigrum, seeds of Foeniculum vulgare,
seeds of Soya, flower buds of clove, seeds of cumin, seeds of cardamom and Floral
part of Myristica fragrans are mixed together at a temperature of from 32 degree
centigrade to 38 degree centigrade.
9. The process as claimed in claim 6, wherein the drying is done at a temperature of
shade drying at 25-35 degree centigrade and followed by drier at 48 degree centigrade
to 58 degree centigrade.
10. The process as claimed in claim 6, wherein dosage range of the composition ranges
from 4.02 gms to 5.94 gms in three divided dose at the minimum interval of 5-6 hrs.