Ray et al.
Formulation, Characterisation and
assessment of Antidiabetic activity of
Polyherbal tablet (PHF) in diabetic rat
Biswaranjan Ray
, B B Panda
and P K Biswal
Diabetes mellitus is a complex and a diverse group of disorders that disturbs the metabolism of carbohydrate, fat and protein.
The number of diabetes mellitus cases has been increasing worldwide in recent years. In 2000, the WHO estimated a total of 171
million of people with diabetes mellitus from the global population, and this report projected to increase to 366 million by 2030.
Now a days polyherbal formulations made by different herbal pharmaceutical company is very much popular and acceptable for
chronic use in case of diabetes, hypertension, bronchial asthma, hyperlipidemia, rheumatoid arthritis etc. As the incidence of
diabetes is increasing day by day globally and the rate of occurrence of disease in India is the high, the popularity of polyherbal
formulation is increasing day by day. Many physicians from different system of medicine refer to advice polyherbal formulations
for long time use in patient of diabetes, as the manufacturer of herbal formulator claim the formulation is having negligible side
effect. Present Study was done with Evaluation of antihyperglycemic Potential of prepared polyherbal Formulation. PHF i.e. with
100mg/kg and 150 mg/kg, the FBS level was reduced from 187.16 ± 3.25 to 133.16 ± 2.93 and 181.5 ± 4.42 to 136.33 ± 2.58,
respectively. In the combination regimen of PHF (150 mg/kg) and Metformin (50mg/kg) the mean FBS level dropped from 180.3
± 1.75 to 131.16 ± 2.14. In all these groups the mean FBS level has reached to the normal pre induction level. The daily single
administration of PHF formulation (150mg/kg) and Metformin (50 mg/kg) significantly reduced blood sugar levels of Streptozotocin
(STZ) induced diabetic rats. It is also important to study antihyperglycemic effect of the said Polyhedral Formulation in different
animal Model to conclude exact pattern of Anti hyperglycaemic activity.
Keywords: Diabetes mellitus; polyherbal formulations; antihyperglycemic; Tamarindus indica; S.rebaudiana
Diabetes mellitus is a metabolic disease characterized by hyper-
glycaemia. It occurs due to either defects in insulin secretion
or insulin action, or both. The chronic hyperglycaemia is re-
lated with damage and malfunction of different organs, predomi-
nantly kidneys, eyes, nerves, blood vessels and heart. Symptoms
Correspondence: bipinbihari030481@gmail.com
Dept. Pharmacology, College of Pharmaceutical Science, Puri, Odisha,
Full list of author information is available at the end of the article.
of marked hyperglycaemia include polyuria, polydipsia, occa-
sionally polyphagia, weight loss and blurred vision. [1]
Over the past 40 years, prevalence of diabetes mellitus has
increased. The trend for future is a continuous increase of all
ethnic groups, men or women, for all age groups worldwide [2].
This increase is observed above all in type 2 diabetes melli-
tus (T2DM) [3]. In 1995, nearly 135 million people were af-
fected and an increase of 300 million cases is estimated for
year 2025 [4]. Various complications of the disease are coronary
artery disease (CAD) including obesity, dyslipidemia, hyperten-
International Journal of Phytomedicine
Received: 13 Dec 2020, Accepted:24 Jan 2021
sion (HT) and physical inactivity [5] . Obesity increases the risk
of CAD in adults and is associated with insulin resistance in nor-
moglycemic as well as in individuals with type 2 DM [6] .
The fruit Tamarind is obtained from the plant Tamarindus in-
dica. The plant is a long lived and large sized tree. It is a famous
as well as common tree found in India. Its fruit is salted and
stored in almost every house. The people of Deccan largely con-
sume it. They say that ‘life is very ticklish in absence of Imli’.
It is also used largely as a flavour, stabilizer and binder in food
preparations. Generally people use Imli to make pickles, curries,
jam and sauce and to prepare majoon as well as jawarish. The
seeds are made flour for making bread in famine seasons. The
seeds are also fried and consumed by poorer. It is very useful for
people of hot area and near to equator.
Tamarind seed powders were used for tannase production by
A. niger. The tannase yield was 6.44 IU/g ds for tamarind seed
According to Ethnobotanical and traditional literature, seeds
act as anti-asthmatic, antiulcer as well as antioxidant agent.
Vaginal Atony is treated by a pessary of seed kernel. The fried
seed paste was used on anus after setting the tract in proper po-
sition for treatment of rectal prolapse. In Unani system, sper-
matorrhoea, nocturnal emissions as well as seminal debility are
cured by use of roasted kernel and seeds. Homeopathic system
uses seed for the treatment of stomachache. It is also used in
cough and for the relaxation of uvula. The urethral discharge and
polyuria are treated by administration of seed kernel pounded
with milk. The red outer covering of seeds is very useful in di-
arrhea and dysentery. The seeds are used for treatment of colitis
and other intestinal disorders. To feed cattle ground seeds are
usually useful. Seeds are helpful in vaginal discharges and ul-
Seeds are rich in phenolic compounds, polymeric tannins,
glycosides, fatty acids, flavonoids, saponins, alkaloids. There is
also presence of fatty acids, essential amino acids in seeds. Aux-
ins are available in seeds. Tamarind seeds have 2-hydroxy-3
, 4
dihydroxyacetophenone (TAO), methy l-3, 4-dihydroxybenzoate
(TA1), 3, 4-dihydroxyphenylacetate (TA2) and (-)-epicatechin.
Seed also consist of acetic acid, arabinose, dihydroxylphenyl
acetate. Oil obtained from seeds consists of lauric, palmitic,
myristic, stearic, arachidic, behenic, lignoceric acids, loeic and
linoleic acids. Dry form of kernel has 17.1-20.1% protein, 6.0-
7.4% fat, 65.1-72.2% carbohydrates and some crude fibre and
ash. Whereas roasted kernel includes calcium 121 mg and phos-
phorous 237 mg per 100 gm. It also contains proteins like pro-
lamines and albumins [4].
For hundreds of years, indigenous peoples in Brazil and
Paraguay have used the leaves of stevia as a sweetener. The
Guarani Indians of Paraguay call it kaa jheé and have used it to
sweeten their yerba mate tea for centuries. They have also used
stevia to sweeten other teas and foods and have used it medici-
nally as a cardiotonic, for obesity, hypertension, and heartburn,
and to help lower uric acid levels.
In addition to being a sweetener, stevia is considered (in
Brazilian herbal medicine) to be hypotensive, diuretic, car-
diotonic, and tonic. The leaf is used for obesity, cavities, hyper-
tension, fatigue, depression, sweet cravings, and infections. The
leaf is employed in traditional medical systems in Paraguay for
the same purposes as in Brazil [5–7].
Lufa cylindrical belongs to cucurbitaceae family having king-
dom Plantae according to botanical classification. The different
plant parts such as leaves, flower and seeds have been used tradi-
tionally for medicinal purpose. The plant part traditionally used
for intestinal worm, Chronic bronchitis, protozoal disease and
for certain kind of fungal infection [8].
Materials and Method
Plant material and Extraction
Stevia (S.rebaudiana) leaves were collected from palisree mela
(local festival) of Western Odisha from a vendor who cultivate
stevia in small scale for commercial purpose.
Lufa cylindrical belongs to cucurbitaceae family having king-
dom Plantae according to botanical classification. The leaves
of the plant were collected from jamadarpli forest area of sam-
balpur district, Odisha.
Stevia leaves were washed to remove dust and subjected to
aqueous extraction after air drying. The dried ground leaves
were mixed with hot water (65 C) at the ratio of 1:45(w/V). The
mixture was shaken and kept at room temperature for 24 hour. It
was sired at least 3-4 times per day. After 24 h,the mixture was
filtered through what man filter paper and the filter was evapo-
rated by using vacume evapoarator.
The leaves of lufa cylindrica were dried in shade and pow-
dered to get a coarse powder. About 800gm of dry coarse powder
was extracted with ethanol (40-60
C) by continuous hot perco-
lation using Soxhlet apparatus. The extraction was continued for
72hours. The ethanolic extract was filtered and concentrated to
a dry mass by using vacuum distillation.
Ether and ethanol extracts of fruits were prepared sequen-
tially by standard procedures in soxhalation apparatus. Matured
unripe fruits were shade dried until properly dried, crushed in a
mechanical grinder into fine powder. The powder (500 g) was
extracted sequentially with 1 litres of ether, 1 litres of ethanol in
a Soxhlet apparatus at 65
C until the powder became exhausted
totally. The resulting extracts were filtered, concentrated and
Ray et al. : International Journal of Phytomedicine, 2021;13(1):005-008
dried in vacuo (yield 7.60, 8.25 and 8.75% w/w, respectively).
The extracts were stored in desiccators for use in subsequent ex-
periments [9].
Preparation of Polyherbal Tablet
The dried Plant extract of all proposed plants were mixed with
different excipients using wet granulation method for preparing
later solid pharmaceutical form. These prepared granules of each
form were compressed into tablet using compressing machine at
Gayatri College of Pharmacy [10].
Characterisation of Polyhedral Tablet
The Prepared Polyherbal Tablet had been undergone characteri-
sation under following parameter such as particle size measure-
ment, colour and appearance, angle of Repose, Wet variation,
Hardness and Friability test were conducted successfully by tak-
ing standard parameters.
Screening of Antidiabetic Potential of Polyherbal formulation
Animals Sprague Dawley (SD) rats of either sex weighing
between 150-200 gm were procured from Scientific Trader,
Baleswar. They were housed in polypropylene cages (six rats
/ cage) and maintained under controlled room temperature (20-
C) and with relative humidity of 45-55% under 12:12hr light
and dark cycle. They were provided with standard lab diet and
water ad libitum and kept for 1 week to acclimatize with the
laboratory condition before starting the experiment.
The rats were grouped into 06 groups and each group has 6
rats. Group 1: considered as control group; Group 2: considered
as treatment group, treated with standard antidiabetic compound,
metformin. Group 3, 4 & 5 each were considered as test group,
treated with polyherbal formulation “PHF” in different doses.
Group 6 consist of 06 rats and treated with combination of both
“PHF” and standard drug “metformin” [11–13].
Acute toxicity study
The acute toxicity study was performed as per OECD (guide-
lines 425) and LD
was calculated accordingly. The animals
were examined at every 30 min up to a period of 3 h and then
occasionally for additional 4 h period, finally 24 h mortality was
recorded. All the animals found to be safe [14].
Experimental induction of Diabetes mellitus
Diabetes mellitus (Type 2 diabetic) was induced in the rats with
normal blood glucose levels by a single injection of streptozo-
tocin (STZ) 35mg/ kg body weight I.P in 0.1M sodium citrate
Table 1 The grouping of rats into different groups for the treatment
of plant extract and the standard drug.
group drug Dose (mg/kg) treatment
I. Normal saline 1ml/kg Control
II. metformin 50.00 Standard
III. PHF 50 Test
IV. PHF 100 Test
V. PHF 150 Test
VI. Metformin +PHF 50+150 Combination
buffer, PH 4.5. In contrast rats in control group were injected
with 0.1M citrate buffer solution only. After injection of STZ
the animals were kept for observation for 48 hrs.
Blood Sample Collection The blood samples were collected
through the tail vein puncturing with a needle. A drop of blood
of size 0.3 to 1.0 µl required. The drop of blood obtained by
pricking with the lancet provided with glucometer for estimation
of FBS and PPBS (Dubois 2014). FBS and PPBS at 1hr and 2hr
were estimated with the help of glucometer weekly for 4 weeks.
The glucometer include a clock that was set for date and time
and memory for past test results. It enables to keep a record of
blood glucose levels over days and wee
Results and Discussion
The mean FBS level of all the rats before induction of diabetics
was found to be 126.5 ± 1.378 mg/dl (Table 1). In contrast the
mean ± S.D of FBS level for different treatment groups over a
time period from week 0 to week 4 was included in Table 2. In
the control group the mean FBS has dropped from 181.16 ± 8.50
in Week 0 to 181.16 ± 2.32. This is far above the mean level
before induction, even after 4 weeks. In Metformin (50mg/kg)
treated group the FBS level was declined from 190.5 ± 2.66 to
138.83 ± 2.23. Similarly in PHF(50mg/kg) treatment group the
FBS was declined from 182.3 ± 6.02 to 135.33 ± 3.01 which
is significant and nearer to the pre induction level. In slightly
higher concentration of treatment of PHF i.e. with 100 mg/kg
and 150 mg/kg, the FBS level was reduced from 187.16 ± 3.25
to 133.16 ± 2.93 and 181.5 ± 4.42 to 136.33 ± 2.58, respec-
tively. In the combination regimen of PHF (150mg/kg) and Met-
formin (50mg/kg) the mean FBS level dropped from 180.3 ±
1.75 to 131.16 ± 2.14. In all these groups the mean FBS level
has reached to the normal pre induction level.
One way ANOVA has been applied for comparison among
different treatments groups and followed by Dunnetts multiple
comparison tests for comparison between pair of treatment ef-
Ray et al. : International Journal of Phytomedicine, 2021;13(1):005-008
Table 2 Descriptive statistics of fasting blood sugar at different time period with different doses of drug treatment
Time Period
Mean FBS ± SD (mg/dl)
Control Metformin
DiaM 50 mg/kg DiaM 100 mg/kg DiaM 150 mg/kg Metformin 50mg/kg
+DiaM 150 mg/kg
Week-0 181.16 ± 8.50 190.5 ± 2.66 182.3 ± 6.02 187.16 ± 3.25 181.5 ± 4.42 180.3 ± 1.75
Week-1 179.13 ± 3.54 144.66 ± 4.50 140.16 ± 1.94 144.5 ± 4.59 141.16 ± 2.86 142.16 ± 3.54
Week-2 179.66 ± 1.75 141.33 ± 2.58 139.33 ± 1.63 139.83 ± 1.94 139.16 ± 1.94 136.16 ± 2.04
Week-3 180.5 ± 1.87 140.5 ± 2.26 133.16 ± 1.83 134.16 ± 1.47 135.16 ± 1.94 133.33 ± 2.66
Week-4 181.16 ± 2.32 138.83 ± 2.23 135.33 ± 3.01 133.16 ± 2.93 136.33 ± 2.58 131.16 ± 2.14
The daily single administration of PHF formulation (150mg/kg)
and Metformin (50 mg/kg) significantly reduced blood sugar
levels of Streptozotocin (STZ) induced diabetic mice. This Anti
hyperglycaemic effect is believed to be due to the ability of
PHF extract to stimulate glucose uptake into cells. Coroso-
lic acid, lagerstroemin, tannic acid and penta-O-galloyl-D-
glucopyranose (PGG) are the known active components in
the leaf extract. Metformin (50 mg/kg) did not cause significant
glucose reduction. Further study in the field of Formulation
needed to overcome manufacturing difficulty as well as making
a suitable herbal solid doses form. It also important to study
antihyperglycemic effect of the said Polyhedral Formulation
in different animal Model to conclude exact pattern of Anti
hyperglycaemic activity.
Conflict of Interest
There is no conflict of interest
Author details
Dept. Pharmacology, College of Pharmaceutical Science, Puri,
Odisha, India.
PG Department, Gayatri College of Pharmacy,
Sambalpur, Odisha, India.
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