INTRODUCTION
Modern lifestyle involves changes in essential
food habits [1]. Due to improper diet, people suffer
from various kinds of chronic diseases. Obesity, heart
disease, and diabetes are among current global issues [2].
Nutraceuticals are food-based drugs which minimize
diet-related illnesses. The word “nutraceutical” comes
from “nutrition” and “pharmaceutical” [3]. Any
pharmaceutical product manufactured from food
sources has fundamental nutritional values and minimizes
various chronic diseases [4].
Recently, nutraceuticals have gained extensive
attention due to their better pharmacological functions
with fewer side effects. In 2017, 77% of Americans
used nutraceuticals, and their number was rising
daily [5]. Nutraceuticals are classified as dietary fiber,
probiotics, prebiotics, polyunsaturated fatty acids,
antioxidant, vitamins, polyphenols, and spices [6].
Herbal nutraceuticals are prepared from plants, fungi,
algae, or their combinations. Various forms of herbal
nutraceuticals (powder, tablets, capsules, liquid, etc.) are
currently available in the market [7].
About 2500 years ago, Hippocrates wrote, “Let
food be thy medicine and medicine be thy food.” This
quotation is undoubtedly the principle of today [6]. The
nutraceutical industry has great potential in some Asian
Foods and Raw Materials. 2022;10(2)
ISSN 2310-9599 (Print)
ISSN 2308-4057 (Online)
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Debnath B. et al. Foods and Raw Materials. 2022;10(2):227–234
countries, such as China, Japan, India, South Korea,
Thailand, Singapore, and Taiwan. With rich natural
resources, skilled human resources, and excellent R&D
facilities, these Asian countries are expected to lead the
production of herbal nutraceuticals [8, 9].
Tripura is India’s state located in the northeastern
part of the subcontinent between 22o7’ and 24o2’ north
latitudes and 91o0’ and 92o0’ east longitudes. Its highest
temperature is 35.60°C, and yearly rainfall is 2000 mm.
These excellent climatic conditions make Tripura a
favorable place for various edible medicinal plants, or
plants with nutritional benefits and bioactive compounds
for the human body [10, 11].
According to literature, the forest of Tripura is a
good source of edible medicinal plants with anti-diabetic
properties [12]. Diabetes is a big issue in the Indian
health care system. Type 2 diabetes is the most common
in adults with uncontrolled diet. Every year, nearly
1 million Indians die due to diabetes. This chronic
disease afflicts 2.8% of the global population [13].
Treatment with herbal medicine improves insulin
secretion and reduces intestinal glucose absorption in
diabetics [14].
This study aimed to formulate herbal nutraceutical
tablets from selected edible medicinal plants of Tripura,
India, and evaluate its anti-diabetic activity using ob/ob
mice (type II diabetes model).
STUDY OBJECTS AND METHODS
Ethnomedicinal data collection. Ethnomedicinal
data about five edible plant parts (Musa paradisiaca
fruit, Musa paradisiaca stem, Cmorphophallus
paeoniifolius corn, Artocarpus heterophyllus seed, and
Colocasia esculenta leaf) were collected from oral
interviews with traditional healers from three ethnic
communities (Tripuri, Reang, and Jamatia) of Tripura.
All the interviewees were adults aged over forty [15].
The scientific names of the collected plant specimens
were identified with the help of a plant taxonomist and
the book, “The Flora of Tripura State” [16].
Processing of plant materials. After identification,
we selected plant parts with medicinal properties,
washed them thoroughly with distilled water, and then
properly air-dried them. The dried samples were ground
to fine powder. The powder material was packed in a
sealed container and preserved at room temperature for
further experiments [17].
Experimental animals. In our experiments, we
used laboratory mice as model organisms. Adult Swiss
albino mice (18–25 g) of both sexes were used for
acute toxicity tests. Male ob/ob mice (type II diabetes
mice weighing 50–55 g, aged 12 weeks) and male
C56BL6 mice (normal mice weighing 28–32 g, aged
12 weeks) were used for an anti-diabetic activity
test. The temperature (20°C) and humidity (53%) of
the animal house were controlled and maintained
throughout the 12 h/12 h light/dark cycle. Food and
water were available except during the fasting period.
The care and handling of the animals were in line with
the regulations of the National Institutes of Health. The
Institutional Ethics Committee (No. 1667/GO/a/12/
CPCSEA) approved the study protocol [18].
Acute toxicity test. Acute toxicity tests of five
different plant powder samples were measured by
the method described by Ali et al. with some modification
[19]. The Swiss albino mice (18–25 g) of both
sexes were divided into two groups, the control group
and the experimental group, ten animals in each. The
control group received only distilled water, whereas
the experimental group received different doses (5, 50,
300, 800, 1200, and 2000 mg/kg body weight) of the
powder sample orally. The mice were then kept under
observation up to 72 h for mortality or symptoms of
toxicity [19].
Formulation of herbal tablet. We used the wet
granulation method to prepare novel herbal antidiabetic
nutraceutical tablets. Equal amounts of
previously prepared plant materials (M. paradisiaca
fruit, M. paradisiaca stem, C. paeoniifolius corn,
A. heterophyllus seed, and C. esculenta leaf) were placed
in a rapid mixture granulator, with a 10% starch solution
added dropwise into the binder. The damp masses
were screened through a sieve #10 and then dried. The
dried granules were screened through a sieve #20 and
stored in a desiccator until they were ready for tablet
compression. The prepared granules were compressed in
a single punch tablet press machine (Manesty Type F3,
Liver Poole, England) with a punch diameter of 0.75 cm
and a compression pressure of 933 Pa (N/m2). The die
volume matched the tablets’ weight to confirm that
600 mg was obtained [20].
Evaluation of granules. Tablet granules were
evaluated by Wadher et al. methods with some
modification. Particularly, we determined their bulk
density, tapped density, Hausner quotient, Carr’s
compressibility index, flow rate, and angle of repose [21].
Evaluation of herbal tablets. Weight variation,
thickness, hardness, friability, and in vitro disintegration
time were determined by Wadher et al. methods with
some modification [21].
Total moisture, total carbohydrate, complete protein,
total fat, total ash, and total caloric value of the herbal
tablets were determined by Debnath et al. method with
some modification [17]. Total dietary fiber was measured
by Ozoliņa et al. method with some modification [22].
Concentrations of minerals were determined by an
atomic absorption spectrophotometer. Debnath et al.
method with some modification was applied to measure
the content of minerals in the tablets [17].
Vitamins C, B1, B3, B6, and folic acid were
determined by Antakli et al. method with some
modification, using the RP-HPLC system [23].
Instrumental conditions:
Column: C18 BDS (10 cm×4.6 mm; 3 μm);
Mobile phase: A = Hexane-1-sulfonic acid sodium
(5.84 mM): acetonitrile (95:5) with 0.1% triethylamine
as solvent (A) at pH 2.5; B = 5.84 mM of hexane-1-
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sulfonic acid sodium: acetonitrile (50:50) with 0.1%
triethylamine as solvent (B) at pH 2.5, pH = 3.54;
Flow rate: 1.6 mL/min;
Injected volume: 20 μL;
Absorbance recorded: Vitamins C and B1 = 246 nm,
vitamin B3 = 260 nm, vitamin B6 = 290 nm, vitamin B9 =
= 282 nm.
Determination of fat-soluble vitamins. Vitamins A,
D3, and E were determined by using the reversed-phase
high-performance liquid chromatography as reported by
Xue et al. with some modification [24].
Instrumental conditions:
Column: dC18 (particle diameter 5 μm, 150×
4.6 mm i.d.);
Mobile phase: methanol:water = 98:2;
Flow rate: 1.00 mL/min;
Injected volume: 10 μL;
Absorbance recorded: vitamin E = 230 nm, vitamin
A and D3 = 265nm.
Anti-diabetic activity. Experimental design. The
animals were randomly divided into four groups of six
animals for test purposes, namely: a) a normal group
(completely healthy mice) treated with 0.5% sodium
carboxymethyl cellulose; b) a vehicle control group
(ob/ob mice) treated with 0.5% sodium carboxymethyl
cellulose; c) a positive control group treated with 200
mg/kg of metformin via gavage; and d) an experimental
group treated with 200 mg/kg herbal nutraceutical
tablet via gavage. The experiment lasted four weeks. At
the end of the experiment, all the animal groups fasted
overnight, and blood samples were collected from the
tail vein. Before blood collection, the animals were
given pentobarbital as an anesthetic agent [18].
Determination of serum hemoglobin and
glycosylated serum protein. Hemoglobin (HbA1c) and
glycosylated serum protein were measured by respective
kits (Merck Millipore, Germany) according to the
manufacturer’s instruction [18].
Oral glucose tolerance test. After four weeks of
treatment with herbal nutraceutical tablets, the animals
were made to fast overnight, and glucose solution
(2 g/kg of body weight) was administered orally. After
that, their blood samples were collected every 30 min
(0, 30, 60, 90, and 120 min). The blood glucose was
measured by a glucose meter (i-QARE DS-W®) [18].
Statistical analysis. For the analysis of granules,
herbal tablets, proximate compositions, minerals, and
vitamins, the data were expressed as mean ± SDs. For
the evaluation of anti-diabetic activity in ob/ob mice,
the data were expressed as mean ± S.E.M. One-way
ANOVA was used to determine significant differences
among groups, after which the modified Student’s
t-test with the Bonferroni correction was applied to
compare individual groups. All statistical analyses
were performed with SPSS 17.0 software. P < 0.05 was
considered statistically significant.
RESULTS AND DISCUSSION
Ethnomedicinal study. The field survey showed
that the five edible medicinal plants were applied by
the healers of Tripura to cure different diseases (Table
1). Ethnomedicinal studies play a vital role in finding
medicinal plants that can be used to produce novel
crude drugs. They also verify the protection of cultural
heritage [11]. The data that we collected from the
informants of healers clearly proved that the plants we
had selected were of medicinal importance.
Acute toxicity test. Our results indicated no changes
of behaviour or mortality caused by the plant samples
at the highest dose of 2000 mg/kg b.wt. This meant
that this dose had no lethal or toxic effect. Toxicity
assessment is one of the crucial steps prior to human
uses of any pharmaceuticals or food ingredients. Acute
toxicity studies determine adverse effects of any active
compound after oral ingestion of a single or multiple
doses [19]. We found that the five plant samples under
study had no toxic effect.
Evaluation of granules. All the physical parameters
of the herbal granules under study were found to be
satisfactory (Table 2). The Carr’s compressibility index
indicates the strength of the powder/granules, while
the Hausner ratio determines the powder/granules’
inter-particulate friction. Both parameters are used
to analyze the powder/granules’ flow rate. The Carr’s
compressibility index of less than 10 or the Hausner
ratio of less than 1.11 indicate an “excellent” flow rate,
while the Carr’s compressibility index higher than 38 or
the Hausner ratio higher than 1.60 indicate a “very poor”
flow rate. Both parameters depend on the bulk density
and the tap density of the powder/granules [25]. The
angle of repose is another parameter used to evaluate
the powder/granules’ flow rate. Its value of less than
30° indicates an “excellent” flow rate, while the value
greater than 56° indicates a “very poor” flow rate [26].
Table 1 Ethnomedicinal use report on edible medicinal plant parts selected
Botanical name and family Local name Plants parts Ethnomedicinal use
Musa paradisiaca (Musaceae) Kola Unripe fruit Diabetes, hypertension, ulcers, diarrhea
Musa paradisiaca (Musaceae) Kola Stem Diabetes, high blood pressure, high acidity
Cmorphophallus paeoniifolius
(Araceae)
Batama Corn Helminths, liver disease, digestive and gastric disorders,
diabetes
Artocarpus heterophyllus
(Moraceae)
Kathal Seed Ulcers, constipation, diarrhea; excessive accumulation
of fluid in tissues
Colocasia esculenta (Araceae) Kocho Leaf Diabetes, microbial infection, liver disease
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Our results showed an excellent flow rate in the granules
prepared with a 10% starch solution (Table 2).
Evaluation of herbal tablets. All the physical
parameters of the herbal tablets under study were found
to be satisfactory (Table 3). The weight variation test is
employed to ensure that each tablet/capsule in the batch
contains the same amount of drug ingredients. Checking
a tablet’s thickness is necessity for packaging since very
thick tablets are not suitable for packaging. A tablet’s
friability and hardness tests are closely related to each
other. They determine the physical strength of a tablet.
Disintegration refers to the mechanical breakup of a
compressed tablet into small granules at a specific time
point. The disintegration test provides critical safety
data on the drug’s bioavailability in the body [27, 28].
In our experiments, 20 tablets were used to check
the average uniformity of weight and 10 tablets to check
the average uniformity of thickness and hardness. For
the friability and disintegration tests, the samples were
prepared in triplicate. According to the results (Table 3),
the physical parameters of our newly formulated tablets
were within the limits established by the United States
Pharmacopoeia.
Determination of proximate composition. The
proximate composition of the herbal nutraceutical
tablets is represented in Table 4. The moisture content is
an essential parameter because high moisture affects the
physical stability of food products. Their shelf life also
depends on the total moisture content [29]. As we can
see in Table 4, our newly formulated herbal tablets had
a low moisture content (62.44 ± 2.11 %), indicating high
physical stability.
Carbohydrates, proteins, and fats hold a special place
in human nutrition. The human body requires them in
relatively large amounts for normal functioning. These
three macronutrients provide energy (measured in
calories) in the human body [30]. Our newly formulated
herbal nutraceutical tablet contained optimal amounts of
carbohydrates (1.88 ± 0.52 %), proteins (0.95 ± 0.65 %),
and fats (0.43 ± 0.53 %), as well as provided a good
amount of energy (15.19 kcal).
So, the herbal nutraceutical tablets developed may
be recommended to people with unbalanced energy
levels. Dietary fiber is a complex mixture of polysaccharides.
Diets with a high content of fiber alleviate
constipation [31]. Our herbal nutraceutical tablet
contained a good amount of dietary fiber (1.66 ± 0.67%),
so it may be used to reduce constipation.
Minerals determination. Iron is an essential
microelement for producing blood. Anemia is the
most common disease caused by iron deficiency.
Every year, approximately 30% of patients suffer from
Table 2 Physical parameters of herbal granules
Physical parameters Herbal granules prepared with
10% starch solution
Bulk density, g/mL 0.46 ± 0.01
Tapped density, g/mL 0.48 ± 0.01
Hausner quotient 1.07 ± 0.01
Carr’s compressibility, % 6.72 ± 0.53
Angle of repose, o 32.29 ± 0.99
Flow rate, g/sec 6.67 ± 0.57
*The values represent mean ± SD for three samples
Table 3 Physical parameters of herbal nutraceutical tablets
prepared with 10% starch solution
Physical parameters Value
Uniformity of weight, mg 610.5 ± 1.70
Uniformity of thickness, mm 3.78 ± 0.041
Hardness, kg/cm2 3.88 ± 0.078
Friability, % 0.65 ± 0.020
Disintegration time, s 295.33 ± 1.52
*The values represent mean ± SD for three samples
Table 4 Proximate composition of herbal nutraceutical tables
Proximate composition Total amount, %
Moisture 62.44 ± 2.11
Carbohydrates 1.88 ± 0.52
Protein 0.95 ± 0.65
Fat 0.43 ± 0.53
Ash 1.78 ± 0.53
Dietary fiber 1.66 ± 0.67
Calories, kcalЦ 15.19
*The values represent mean ± SD for three samples
Table 5 Minerals profile of herbal nutraceutical tablets
Minerals Total amount, mg/g
Iron 74.6 ± 2.7
Sodium 4.4 ± 0.4
Potassium 5.3 ± 0.7
Calcium 163.1 ± 2.2
Magnesium 39.2 ± 1.7
Phosphorus 14.6 ± 2.1
*The values represent mean ± SD for three samples
Table 6 Profiles of water soluble and fat-soluble vitamins in
herbal nutraceutical tablets
Vitamin Total amount
C (ascorbic acid), mg/g 27.2 ± 4.3
Vitamin B1 (thiamin), mg/g 0.6 ± 0.0
Vitamin B3 (niacin), mg/g 0.6 ± 0.2
Vitamin B6 (pyridoxine), mg/g 1.1 ± 0.2
Vitamin B12 (cobalamin), mcg/g 0.6 ± 0.2
Vitamin B9 (folic acid), mcg/g 82.6 ± 7.6
Vitamin A (retinol), mcg/g 287.4 ± 6.3
Vitamin D3 (cholecalciferol), mcg/g 2.6 ± 0.6
Vitamin E (tocopherol), ng/g 0.7 ± 0.0
*The values represent mean ± SD for three samples
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anemia globally, particularly 51% in India [32]. Our
herbal nutraceutical tablet had a significant amount
of iron (74.6 ± 2.7 mg/g), so it may alleviate the
effects of iron deficiency (Table 5). Dietary potassium
and sodium are two electrolytes that play a vital
role in regulating fluid and blood volume [33]. The
tablets developed contained a fair amount of sodium
(4.4 ± 0.4 mg/g) and potassium (5.3 ± 0.7 mg/g),
so it may be used to regulate the body’s fluid
and blood volume. Calcium, phosphorus, and
magnesium are three crucial micronutrients for
healthy bone and teeth formation, as well as
metabolic functions [34]. Our herbal nutraceutical
tablets contained a considerable amount of
these minerals, namely 163.1 ± 2.2, 14.6 ± 2.1,
and 39.2 ± 1.7 mg/g, respectively. Therefore, the newly
developed herbal tablets can help bone formation and
improve metabolic functions in the human body.
Determination of vitamins. We determined the
values of water-soluble (C, B1, B3, B6, B12, and folic
acid) and fat-soluble vitamins (A, D3, and E) in our
herbal nutraceutical tablets (Table 6). We found rich
amounts of vitamin C (27.2 ± 4.3 mg/g) and vitamin A
(287.4 ± 6.3 mcg/g). Vitamin C is a water-soluble
vitamin that works as an antioxidant and improves the
immune function of the human body [35]. Vitamins B1,
B3, B6, and B9 are essential for maintaining the nervous
system, digestion, protein metabolism, red blood cells,
Figure 1 Carbohydrate metabolism parameters of ob/ob mice treated with sodium carboxymethyl cellulose (normal group), sodium
carboxymethyl cellulose (vehicle control group), metformin (positive control group), and herbal nutraceutical tablet (experimental
group). Each group contained six animals
c d
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Figure 2 Change of blood glucose with time in ob/ob mice
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sodium carboxymethyl cellulose (vehicle control group),
metformin (positive control group), and herbal nutraceutical
tablet (experimental group). Each group contained six animals
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15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
HbAlc, ng/mL
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
HbAlc, ng/mL
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
0
5
10
15
20
25
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
##
## ** ***
0
5
10
15
20
25
30
35
##
*** ***
#
0
5
10
15
20
25
30
35
40
**
##
## ***
0
5
10
Normal Vehicle Metformin Herbal tablet
Blood glucose, #
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
10
15
20
25
30
35
Normal Metformin tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Herbal tablet
Blood glucose, ##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
Normal Vehicle Metformin Herbal tablet
Blood 0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
5
10
15
20
25
30
35
Normal Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
Normal Vehicle Metformin Herbal tablet
Blood 0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
5
10
15
20
25
30
35
Normal Metformin ##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
Normal Vehicle Metformin Herbal tablet
Blood 0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
0
5
10
15
20
25
30
35
Normal Metformin Herbal tablet
HbAlc, ng/mL
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
0
5
Normal Vehicle Metformin Herbal tablet
Blood 0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
Normal Vehicle Metformin Herbal tablet
Blood 0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
0.5
1.0
1.5
2.0
Normal Vehicle Metformin Herbal tablet
Blood glucose during 4 weeks, ## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Vehicle Herbal tablet
Glycosylated protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
25
Normal Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
5
10
15
20
25
Normal Vehicle Metformin Herbal tablet
Blood glucose, mM
##
*
#
*
0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Normal Vehicle Metformin Herbal tablet
Blood glucose change
during 4 weeks, mM
##
## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0.5
Vehicle Metformin Herbal tablet
during 0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## 0
0.5
1.0
Normal Vehicle Herbal tablet
Blood during ## ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/**
##
## ***
0
0.5
1.0
Normal Vehicle Metformin Herbal tablet
Blood during ## 0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0.5
1.0
Normal Vehicle Metformin Herbal tablet
Blood during ## 0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0.5
1.0
Normal Vehicle Metformin Herbal tablet
Blood during ## 0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
0
***
0
0.5
1.0
Vehicle Metformin Herbal tablet
Blood during ## ** ***
0
5
10
15
20
25
30
35
Normal Vehicle Metformin Herbal tablet
##
*** ***
HbAlc, ng/mL
#
0
5
10
15
20
25
30
35
40
Normal Vehicle Metformin Herbal tablet
Glycosylated serum protein,
ng/mL
**
##
## ***
Normal
232
Debnath B. et al. Foods and Raw Materials. 2022;10(2):227–234
type 2 diabetes [39]. The oral administration of the
herbal tablet to ob/ob mice significantly reduced their
blood glucose level (Fig. 2). Therefore, this tablet may
be used as an anti-diabetic drug.
We also found that the herbal nutraceutical tablet
(200 mg/kg) significantly enhanced oral glucose
tolerance (Fig. 2). This means that this tablet can be used
to alleviate type 2 diabetes mellitus through progressing
insulin sensitivity.
CONCLUSION
Our study showed that the newly formulated herbal
tablet contained optimal amounts of macro- and
micronutrients, water, and fat-soluble vitamins. The
tablet also provided significantly higher hypoglycemic
activity compared to the standard drug, metformin.
The results suggested that the herbal tablets developed
may be recommended as an anti-diabetic herbal
remedy.
CONTRIBUTION
The authors were equally involved in writing the
manuscript and are equally responsible for plagiarism.
CONFLICT OF INTEREST
The authors have declared no conflict of interest in
relation to this manuscript.
and skin health. Vitamins A, D3, and E are common
fat-soluble vitamins that support such body functions
as vision, hair growth, bone maintenance, immune
system regulation, oxidative stress prevention, etc. [36].
Our herbal nutraceutical tablets contained fair amounts
of vitamins D3, E, and B group (Table 6). Therefore,
they may be used to treat diseases caused by their
deficiency.
Anti-diabetic activity. The treatment of ob/ob
mice with our herbal nutraceutical tablets (200 mg/kg
body weight) significantly decreased their fasting blood
glucose, serum hemoglobin, and glycosylated serum
protein (Fig. 1). The effectiveness of the herbal tablet
was comparable to that of metformin, a standard drug
(200 mg/kg body weight).
A fasting blood glucose test, which is generally
called a fasting plasma glucose test, measures the
amount of glucose in the blood and determines the
patient’s risk of prediabetes or diabetes [37]. Our
newly formulated herbal tablet (200 mg/kg) reduced
the fasting blood glucose level (Fig. 1a and 1b).
Serum hemoglobin and glycosylated serum protein
are important indicators for the long-term glycemic
control [38]. Our experiment showed that the herbal
tablet (200 mg/kg) reduced their levels, compared to
metformin (200 mg/kg) (Fig. 1c and 1d).
An oral glucose tolerance test determines the body’s
response to glucose. This test can be used to detect

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