A

Project report
on

Formulation And Evaluation of Polyherbal Cookies

For Diabetic Patient

Submitted in partial fulfillment course of

the requirement for the Degree of

Bachelor of Pharmacy

Submitted by

Vishal Kumar
(B. Pharm VIII Sem)
Reg.no.21109168017

Guided by
Assistant Professor.Rajnish
Kumar Kushwaha

RAJENDRA KISHORI PHARMCY COLLEGE,

Sughari,Siwan (Bihar) 841406
 RAJENDRA KISHORI PHARMACY
COLLEGE,Sughari,Siwan (Bihar) 841406
2021-2025

CERTIFICATE
This is to certify that the Project work presented in this thesis entitled “Formulation
And Evaluation of Polyherbal Cookies For Diabetic Patient’’ is the project of
Vishal Kumar conducted in Rajendra Kishori Pharmacy College Sughari,Siwan
(Bihar), under supervision of Assistant Professor.Rajnish Kumar Kushwaha. A
project work submitted in partial fulfillment of the requirement for the degree of
Bachelor of Pharmacy
in the faculty of Science and Technology, Aryabhatta Knowledge University,Patna

This Project work is now ready for examination.

Uttpal Pandey

Principal

Place: S i w a n
Date:
 RAJENDRA KISHORI PHARMCY
COLLEGE,Sughari,Siwan (Bihar) 841406

2021-2025

CERTIFICATE

This is to certify that the Project work presented in this thesis entitled “
Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient’’ is the project
of Vishal Kumar.conducted in R a j e n d r a K i s h o r i P h a r m c y C o l l e g e ,
S u g h a r i , S i w a n ( B i h a r ) 8 4 1 4 0 6 , under my supervision. The Project work
submitted in partial fulfillment of the requirement for the degree of
Bachelor of Pharmacy in the faculty of Science and Technology, Aryabhatta
Knowledge University, Patna.

This Project work is now ready for examination.

Signature Of The Guide

Assistant professor.Rajnish
Kumar Kushwaha

Place: S i w a n
Date:
 HI-TECH COLLEGE OF PHARMACY
Padoli Phata, Nagpur highway, Morwa, Chandrapur
2022-2023

DECLARATION

I V i s h a l K u m a r , hereby declare that the Project work presented in

this thesis entitled “Formulation And Evaluation of Polyherbal Cookies For
Diabetic Patient’’ was carried out in group under the supervision of A s s i s t a n t
P r o f e s s o r . R a j n i s h K u m a r K u s h w a h a , conducted in R a j e n d r a
K i s h o r i P h a r m a c y C o l l e g e , S u g h a r i , S i w a n . A Project work
submitted in partial fulfillment of the requirement for the degree of Bachelor of
Pharmacy in the faculty of Science and Technology, Aryabhatta Knowledge
University,Patna.

This Project work is now ready for examination and evaluation.

Vishal Kumar

(B. Pharm VIII Sem)

Reg.no.21109168017

Place: Siwan

Date:
 Acknowledgement

It is a moment of great pleasure and immense satisfaction for us to express our respect
and reverence to R a j e n d r a K i s h o r i P h a r m c y C o l l e g e , S u g h a r i ,
S i w a n ( B i h a r ) , for giving us the opportunity to obtain the degree of Bachelor of
Pharmacy.

we convey our deep gratitude and respect to our esteemed Guide, Assistant Professor.
Rajnish Kumar Kushwaha , R a j e n d r a K i s h o r i P h a r m c y
C o l l e g e , S u g h a r i , S i w a n ( B i h a r ) , who has been a continuous
source of inspiration throughout the course of our work. Her deep
involvement and logical thinking has made high difference towards our Project work.

Our special thanks go to U t t p a l P a n d e y , Principal, Rajendra Kishori

Pharmcy College, Sughari, Siwan( Bihar),for providing us facilities for my Project
Report work to be conducted smoothly. Without his valuable support this work was
untouchable for us.

Words are not enough to express our deepest sense of thankfulness to our friends for
their constant help and support. Friendship all about trusts each other, helping each
other, loving each other and being crazy together.

Our family is the persistent source of inspiration for us, whose blessings are always
with us and which always helped us to move smoothly onto the road of success. Our
real sources of inspiration are Our Parents who come before God for us.

At last but not least, we want to extend our thanks to all those who directly or indirectly
helped us for completion of this Project Report.

I extend our deepest sense of gratitude to God for blessing.

 INDEX

Sr.No. Content Page No.

Abstract 1

1. Introduction 2-3

2. Literature review 4

3. Aim and objectives 5

4. Drug Profile 6-16

5. Plan of work 17

6. Material And Methods 18-19

7. Evaluation Parameters 20-24

8. Result and discussion 25-27

9. Conclusion 28

10. References 29-30

List of Figures:

Sr.no Name of figures Page no.

1 Tecoma stans leaves 7
2 Tecoma stans bark 7
3 Withania somnifera 12
4 Avena sativa L 14
5 Preparation of cookies 19
6 Acceptance of taste 26
7 Acceptance of flavor 26
8 Comparative studies with other marked product 27

List of Table:

Sr.no Name of tables Page no.

1 List of ingredients used 18
2 List of equipment used 18
3 Preliminary batches for polyherbal cookies 19
4 Chemical and physiochemical parameter 25

List of Abbreviations:

MCF-7 Michigan Cancer Foundation-7

PIP3 Phosphatidylinositol (3,4,5)-trisphosphate
PI3 Phosphoinositide 3-kinases
GLUT-4 Glucose transporter type 4
METS Methanolic extract of tecoma stans
DPPH 2,2-diphenyl-1-picrylhydrazyl
FBS Fetal bovine serum
MEF Mouse embryo fibroblast
DDSS Diabetes distress scale score
LB TEST Ljung-Box Test
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

ABSTRACT
Most of the people consume cookies during their breakfast, snacks and leisure time to regulate
their hunger and get some energy, in market there are varieties of cookies which are available, and
the main components are refined flour, sugar and butter. Hence, are generally avoided by obese
and diabetic patients as they lead to high sugar level in blood. Therefore, in this recent
investigation, we have formulated Polyherbal cookies using oats, wheat flour and different
Ayurveda herb. Diverse varieties were formulated using different plant to find out the best
composition for cookies on the basis of palatability. After selection, cookies were formulated for
the physiochemical, sensory and their nutritional analysis. Sensory analysis was evaluated based
on organoleptic characteristics: color, taste, aroma and overall acceptability on the basis of 9-
paledonic scale. Physiochemical evaluation including: total ash value, total moisture content total
water and alcoholic extraction, total moisture content. On the basis of its nutritional value
comparison, it was found that the protein content is higher in our formulation than the other
marketed preparation.

Rajendra Kishori Pharmacy College,Sughri Siwan (Bihar) 841406 1

 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

1. INTRODUCTION
Plants have provided a source of inspiration for novel drug compounds, as plant derived medicines
have made large contributions of human health and well-being. Traditional medicine using plant
extract continues to provide health coverage for over 80% of the world’s population, especially in
the developing world. Plants are used medicinally in different countries and are a source of many
potent and powerful drugs.

Medicinal plants provide good remedies for human diseases and play a vital role in our day-to-day
life. Indian systems of medicines are all based on the knowledge of drugs from plants. Higher
plants, as sources of medicinal compounds, have continued to play a dominant role in the
maintenance of human health since ancient time. Over 50% of all modern clinical drugs are natural
product origin and natural products play an important role in drug development programs in the
pharmaceutical industry. In India people have been used plants and natural products for the
treatment of various diseases from ancient time. In the last two decades of the century the scientists
are sincerely trying to evaluate many plant drugs used in traditional system of medicine. [1]

Diabetes- common, non-communicable metabolic disorder mostly occurring in young ones and
are associated with other diseases like kidney disease, cardiovascular diseases etc. It occurs when
pancreas does not produce insulin or is not properly consumed by body. According to the WHO
survey report globally, it is estimated that 422 million adults were suffering in 2014 and rate of
increment is about from 4.7% to 8.5%. Diabetes mellitus, a chronic metabolic disease condition
enounces by the elevated glucose level in blood (Hyperglycemia), hyperlipidemia, negative
nitrogen balance, glycosuria and sometimes ketonemia. Basically, it is grouped in three main
types: Type-1 diabetes mellitus / Insulin dependent diabetes mellitus/Juvenile onset diabetes
mellitus, which are mostly caused by the T-cell mediated autoimmune devastation of islets of
insulin secretion Beta cells. Type-2diabetes mellitus /Non-Insulin Dependent diabetes mellitus
occurs due to the moderate reduction in β cell mass or the low circulation of insulin due to the
interaction of genetic or metabolic factor. Nearly 90% of diabetes patients are suffering from type-
2 diabetes. And Type-3 occurred during pregnancy when insulin is not produced properly or the
cells become resist to insulin.

Rajendra Kishori Pharmcy College, Sughari, Siwan(Bihar) 841406

.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Insulin is a hypoglycemic hormone with two polypeptide 51amino acid chains. 1U insulin is
secreted per hour by the human pancreas but maximum amount is being secreted after meal and
are controlled by chemical, hormonal and neural mechanism. Insulin is site specific and is present
on all cell membrane but more densely depends on the cell type: liver and fat cells are rich in its
presence. Some of the secondary messenger like PIP3 activate PI3 kinase also facilitates the action
of insulin on metabolism enzymes. Insulin activates glucose transport through the cell membrane
by ATP dependent transfer of glucose transporter (GLUT-4) to plasma membrane.

The secondary messenger PIP3 and certain tyrosine phosphorylated guanine exchanger protein
help GLUT-4 to move from cytosol to the plasma membrane. Recent drugs which are used to
treat diabetes worldwide are in effect, but some natural therapies, natural foods and various
changes in the lifestyle would be a wise act to treat and control diabetes and its associated disease.
Various plants, its part or products (more than 1200) are identified as hypoglycemic with fewer
side effect. [2]

Polyherbal antidiabetic cookies are the combination of various herbs and cereals that have a great
power to stimulate insulin secretion by countless mechanism of action. These cookies contain oats,
Tecoma stans leaves, Tulsi leaves, ashwagandha, honey, artificial sugar (sucralose) etc., which is
tasty,healthy as well as beneficial for a diabetic patient of all age group and as the product contains
herbs and cereals it has less adverse effect on ones body. According to WHO report, 2016 it was
made an action plan for the year from 2013-2020 in which it involved various action plans and
suggested natural food products which can decrease or regularise the blood glucose level in blood
should be develop more. [3]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

2. LITERATURE REVIEW
2.1 A Complete Review on Avena sativa
Kinthali Usha Rani et al, (2021),
The review of all clinical results study Avena sativa a potential candidate as nutraceutical &
pharmaceutical agent. The advantages seen within clinical studies, but it must be well tried in
larger studies for far exploration within therapeutic aspect would be helpful.

2.2 Evaluation of the Cookies Formulated with Costus igneus Plant Material for Antidiabetic
Activity
A Rajani Chowdary et al, (2020),
The present research work aimed to formulate a nutritionally rich cookie with Costus igneus
leaf extract and to determine the effect of cookie consumption on decreasing blood glucose
level in type 2 Diabetic patient.

2.3 Therapeutic Properties and Applications of Tecoma stans Linn.

Abisha Vince Jeo V.S et al, (2020),
In the recent study, the researcher gave the evidence that it contains chemical constituents like
alkaloids, amino acids, phytosterols, monoterpenes, triterpenes, glycosides and phenols,
tannins, saponin & flavonoids. The present review comprises the pharmacological,
phytochemical & therapeutic potential of Tecoma stans.

2.4 Formulation of Polyherbal Antidiabetic Cookies

Kajal Pathak et al, (2018),
The cookies are generally avoided by obese and diabetic patients as they cause high sugar level
in blood. So, objective of present research was to formulate polyherbal Cookies with the help
of oats, wheat flour & different Ayurvedic Herbs.

2.5 Phytopharmacology of Ashwagandha as an Anti-Diabetic Herb

 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

3. AIM AND OBJECTIVE

Aim
The aim of the present study is to design and formulate a cookie formulation as a drug delivery
system for diabetic patients.

Objective
 The objective of the study is to design polyherbal cookies containing Tecoma stans, Tulsi leaves
and ashwagandha.
 The essential target of the study was to formulate and evaluate antidiabetic polyherbal cookies that
will have to work on antidiabetic enhancing property.
 Polyherbal formulation has wide therapeutic range, pure side effects, cheaper eco-friendly and
rapidly available
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

4. DRUG PROFILE
Oats (Avena sativa) belongs to family Poaceae. It has soluble fibre known as beta glucan is
polysaccharides which containing glucose residue binds with 1, 3 and 1, 4 bonds. Products that
have β glucan have been used from thousands of years for human health, but β glucans has been
recently scrutinized as active ingredient. Since then, they have been studied extensively for immune
stimulation effects and developed for the treatment of various diseases including cancer, infectious
diseases. Oat β glucan have been used for several clinical trials to reduce glucose. Studies showed
that oat β glucan lowered postprandial glycemia. The effect of β glucans to reduce blood glucose
could be mediated possibly by delaying stomach emptying so that dietary glucose is absorbed more
gradually. After ingestion of oats (bran flour or crisp), the blood glucose level was lower at 15, 30
and 45 min but higher at 90 min after 12.5g glucose loading. Thus, the peak level is much smoothed
and the shape of the plasma glucose response curve is flatter. These changes reduce the feeling of
hunger caused by rapid decrease in blood glucose. Thus, β glucan may reduce appetite and reduce
food intake. [4]

Ashwagandha (Withania somnifera) belong to family Solanaceae. Ashwagandha root is mainly

containing steroids and alkaloids that have rejuvenative herb that is used to treat mental health
problem associated with other disease. It controls the lipid profile; cholesterol level and blood
glucose level as compare to oral hypoglycemic agent. The flavonoids and antioxidant which is
present in root of Ashwagandha is responsible for its antidiabetic property through improve the
liver and kidney function and balanced albumin and globulin ratio that play a vital role to treat
diabetes. [5]

Tecoma stans is commonly planted as an ornamental in warmer climates throughout the world
because of its showy yellow flowers and pinnate foliage. Tecoma stans is an important medicinal
plant. The major bioactive compounds like alkaloids, phenols, terpenoids, glycosides, flavonoids,
saponins had been isolated from this plant. The leaves, bark and roots contain biologically active
chemicals, and extracts from those tissues are in use as traditional folk medicines. The presence of
phytoconstituents like phytosterol, triterpene, glycosides, phenols, flavonoids, saponins, and
tannins either individually or combined together may exhibit the synergistic effect towards healing
of wounds. [6]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Figure 1. Tecoma stans leaves Figure 2. Tecoma stans bark

4.1 Scientific name: Tecoma stans(L) kunth.

Taxonomy:
Kingdom Plantae – plantes, Planta, Vegetal, plants
Plantae
Subkingdom Viridaeplantae – green plants
Infrakingdom Streptophyta – land plants
Division Tracheophyta – vascular plants, tracheophytes
Subdivision Spermatophytina – spermatophytes, seed plants, phanerogams
Infradivision Angiospermae – flowering plants, angiosperms, plantas com
flor, angiosperma, plantes à fleurs, angiospermes, plantes à fruits
Class Magnoliopsida
Superorder Asteranae
Order Lamiales
Family Bignoniaceae – bignonias
Genus Tecoma Juss. – trumpetbush
Species Tecoma stans (L.) Juss. ex Kunth – yellow elder, yellow
trumpet-flower, trumpet bush, trumpet flower.

Description
Tecoma stans is a promising species in the trumpet vine family, Bignoniaceae. The common names
are yellow trumpet bush, yellow bells, yellow elder and Esperanza. It is a flowering perennial shrub
or small tree, 5-7.6 m in height. Bark is pale brown to grey and roughens with age. Leaves are
opposite, compound and imparipinnate with 2 to 5 pairs of leaflets and a larger single terminal
leaflet.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Flowers occur in clusters at the ends of the branches and are trumpet shaped with 5 rounded lobes,
6 cm long, pale to bright yellow, with faint orange stripes at the throat. Stamens 4, attached at
summit of tube in 2 unequal pairs, filaments pilose at base, curved above, anthers versatile, linear,
yellow, pilose and 6 mm long; Fruits are narrow, slightly flattened to pointed capsules, up to 20 cm
long, containing many winged seeds; green when young, pale brown on ripening and remain on the
tree in untidy clusters for many months. [7]

Phytochemical properties
Tecoma stans leaves bark, and roots contain many biologically active chemicals, and extracts from
those tissues have been used in traditional folk medicine to treat many diseases and conditions. The
major bio active compounds like alkaloids, phenols, terpenoids, glycosides, flavonoids, saponins
have been isolated. These bioactive compounds contain various phytoconstituents. The
phytochemical screening of the Tecoma stans plants varied according to the solvents used for the
extraction of the leaves. Therapeutically important active principle of Tecoma stan is Tecomine
(the alkaloids isolated from the plant harvested in Egypt) was shown to be one of the compounds
responsible for the hypoglycemic action given the interest in substances able to treat type II
diabetes. The Tecoma stans leaves and barks both shows the anti-inflammatory activity. The
biosynthesis of monoterpene alkaloids in callus tissues of Tecoma stans has been studied, together
with the identification of the presence of lapachol and other primary and secondary plant
metabolites such as: sugars (glucose, fructose, sucrose and xylose), triterpenoids (ursolic and
oleanolic acids and α- amyrine), p-sitosterol and phenolics (chlorogenic, caffeic, vanillic, o-
ceramics and sinapicnacids). Alkaloids namely 5β Hydroxyskitanthine, early called Base C, and
boschniakine were inactive both in vivo and in vitro assays. Other chemical constitutes are
phytosterols, alkaloids, quinones, amino acids, monoterpenes, triterpene, glycosides, phenols,
flavonoids, saponins, and tannins. Recently the presence of iridoid glycosides, indolic compounds
in the leaves has been identified from this plant. [8]

Pharmacological activities
Tecoma stans have various pharmacological activities like antidiabetic, anti-inflammatory, wound
healing, antimicrobial, antioxidant, cytotoxic, antifungal and anti-proliferative activity.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

 Anti-Diabetic activity
Antidiabetic effect of Tecoma stans was attributed to alkaloids denominated tecomine and
tecostanine. It has been reported that sub chronic and acute administration of the alkaloid tecomine
diminished plasmatic cholesterol and triglycerides levels without modifying fasting glucose.
Tecoma stans aqueous extract of leaves exert its antidiabetic effects by stimulating glucose uptake
in both insulin-sensitive and insulin-resistant murine and human adipocytes without significant
proadipogenic or antiadipogenic side effects.[9]

In another study it has been reported that the ethanolic extract of Tecoma stans stem (200 mg/kg)
showed statistically more significant in decrease blood glucose level. The ethanolic extract shows
(147.5±4.4) more significant value, when compared to positive control group and near to standard
group (124.6±3.9). The antidiabetic activity of ethanolic extract of Tecoma stans stem may be due
to potentiation of insulin secretion from β-cells of pancreas. The potential antidiabetic activity
may be due to the actions of phytochemicals such as flavonoids, saponins and alkaloids present in
ethanolic extract of Tecoma stans stem. [10]

 Anti-Inflammatory activity
The methanol, ethanol and water extracts of T. stans showed anti-inflammatory activity by
inhibiting the heat induced albumin denaturation and red blood cells membrane stabilization. In
another studies administration of alcohol extract at 250 and 500 mg/kg inhibited the edema from
the 3h after carrageenan challenge, and aqueous extract at dose of 250 and 500 mg/kg inhibited
the edema from 4h after carrageenan challenge, which probably inhibits the different aspects and
chemical mediators of inflammation. [11]

 Wound healing activity

Wound healing process is a complex series of events that begins at the moment of injury and can
continue for months to years. The stages of wound healing are inflammatory phase, proliferation
phase, fibroblastic phase and maturation phase. Different extracts of the bark of Tecoma stans was
evaluated for its wound healing potential in two different types of wound models in albino rats.
The methanolic extract of Tecoma stans (METS) leaf shows marked reduction in the wound area,
significant increase in wound contraction and formation of scar in excision wound model when
compared to controls. This activity is attributed to the presence of phytoconstituents like
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

phytosterols, triterpenes, glycosides, phenols, flavonoids, saponins, and tannins either individually
[12]
or combined together may exhibit the synergistic effect towards healing of wounds.

 Antimicrobial Activity
The alcoholic and aqueous extract of T. stans exhibited the antibacterial activity and the growth of
E. coli and B. subtilis was inhibited at different concentrations. The methanol extracts of leaves and
stem bark of Tecoma stans was studied for their antimicrobial activity using a wide range of Gram-
positive and Gram-negative bacteria and fungi. It was observed that the extracts of stem bark
generally showed better antimicrobial activity than those of the leaves. In another study, the
ethanol, methanol and water leaf extracts of Tecoma stans was tested on various bacteria
(Pseudomonas fluorescens, Clavibacter michiganensis, Staphylococcus aureus, E. coli,
Pseudomonas aeruginosa and Klebsiella pneumonia) and was found to be effective. Phytochemical
analysis revealed the presence of alkaloids, flavonoids, saponins, phenols, steroids, anthraquinones
and tannins.

The three extract fractions have showed highest total phenolic content (177-216mg gallic acid
equivalent/g) which may be attributed to its antimicrobial activity. Flavonoids also have been
proven to display a wide range of pharmacological and biochemical actions, like antimicrobial
activities. [13]

 Antioxidant Activity
Plants are primary resources for antioxidants like carotenes, phenolic acids, etc., the phytochemical
analysis 0f Tecoma stans revealed that the plant has alkaloids, steroids, glycosides and
carbohydrates. Flavonoids can act as free radical scavengers and terminate the radical chain
reactions that occur during the oxidation of triglycerides in the food systems. [14]

The presence of tannins in the extracts may explain its potent bioactivities known to possess potent
antioxidants. The antioxidant activity of methanolic and ethanolic extract of Tecoma stans plants
parts was measured by the capability of DPPH free radicles scavenging in comparison to the
standards, ascorbic acid and butylated hydroxytoulene. DPPH assay results show that methanolic
extract of T. stans in higher concentration show better antioxidant potential than the standard L-
ascorbic acid. They exhibited strong antioxidant potential at 20 μg/mL concentration. [15]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

 Cytotoxic Activity
Cytotoxic effects of Tecoma stans were found to be concentration and time-dependent in the
presence and absence of fetal bovine serum (FBS). Cytotoxicity was determined
spectrophotometrically by MTT and reported in terms of % cell viability. In another study the
cytotoxicity effects of different concentrations of alcoholic and aqueous extracts 100000(A),
50000(B), 25000(C), 12500(D), 6250(E), 3125(F), 1562.5(G), 781.2(H) μg mL-1 were tested on
normal mouse embryo fibroblast cell line (MEF) and treated cell line. The higher concentrations of
the plant extracts had a significant cytotoxic effect than lower concentrations on the MEF cell line.
Morphological assessment using inverted microscopy of the cultured cells indicated that cell death
occurred in the treated cell lines. The active constitutes in the plant extract causes morphological
changes and cell death. [16]

 Antifungal activity
Antifungal activities were tested by the drop diffusion method. Tecoma stans differ significantly in
their activities against the microorganisms tested and most of the extracts showed antifungal
activity against Candida albicans, Cryptococcus neoformans and Microsporum gypseum. Tecoma
stans found to give the best zone of inhibition against the fungal activity. In another study Tecoma
stans was tested against two species of subcutaneous fungi: Sporothrix schenckii and Fonsecaea
pedrosoi by the agar dilution method at a concentration of 100 μ/mL. Tecoma stans, showed
effective activity against F. pedrosoi at MIC 12.5 μg/mL. [17]

 Anticancer activity
Natural products derived from medicinal plants have gained significance in the treatment of
cancer. Compounds which are derived from plants one or another way constitutes more than 50%
of antiproliferative agents. Traditionally flowers and bark of Tecoma stans are used for treatment
of various cancers. Tecoma stans leaves and bark shows anti proliferative activity. Report
suggested that the hydroxyl groups of the aromatic rings (acteoside) appear to play an essential
role in the antiproliferative effect. A novel monoterpene alkaloid, 5- hydroxy-skytanthine
hydrochloride isolated from Tecoma stans Juss. fruits and flowers are effective as an anti-
proliferative against MCF-7 and as a NO inhibitor. The invitro anti- proliferative activity of the
various parts of the Tecoma stans was studied in the Breast cancer. MCF-7 cell lines by MTT
assay. The stem, root, bark and flower extracts showed significant anti-proliferative action on the
cell line (MCF-7) but extreme action was found to be in extract stem bark of Tecoma stans.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Another study shows a dramatic invitro anticancer activity of ethanolic leaf extract of Tecoma
stans on human breast cancer cell line (MCF-7) at increasing concentrations with Inhibitory
concentration 64.5μg/ml. [18-19]

4.2. Scientific Name: Withania somnifera (WS)

Taxonomy:
Kingdom Plantae, Plants
Subkingdom Tracheobionta, Vascular plants
Super division Spermatophyta, Seeds plants
Division Angiosperma
Class Dicotyledons
Order Tubiflorae
Family Solanaceae
Genus Withania
Specie Somnifera dunal

Figure 3. Withania somnifera

 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

It is in use for a very long time for all age groups and both sexes and even during pregnancy without
any side effects. Historically, the plant has been used as an antioxidant, adaptogen, aphrodisiac,
liver tonic, anti-inflammatory agent, astringent and more recently to treat ulcers, bacterial infection,
venom toxins and senile dementia.

Clinical trials and animal research support the use of WS for anxiety, cognitive and neurological
disorders, inflammation, hyperlipidaemia and Parkinson’s disease. WS chemo preventive
properties make it a potentially useful adjunct for patients undergoing radiation and chemotherapy.
Recently WS is also used to inhibit the development of tolerance and dependence on chronic use
of various psychotropic drugs.

WS is a small, woody shrub in the Solanaceae family that grows about two feet in height. It can be
found growing in Africa, the Mediterranean, and India. An erect, evergreen, tomentose shrub, 30-
150 cm high, found throughout the drier parts of India in waste places and on bunds. Roots are
stout fleshy, whitish brown; leaves simple ovate, glabrous, those in the floral region smaller and
opposite; flowers inconspicuous, greenish or lubrid-yellow, in axillary, umbellate cymes; berries
small, globose, orange-red when mature, enclosed in the persistent calyx; seeds yellow, reniform.
The roots are the main portions of the plant used therapeutically. The bright red fruit is harvested
in the late fall and seeds are dried for planting in the following spring. Parts used: Whole plant,
roots, leaves, stem, green berries, fruits, seeds, bark are used. [20]

Pharmacological Activity
Antidiabetic Activity
The use of Ashwagandha is also considered in the context of anti-diabetic effects. However, there
are relatively few reports on this issue. The antidiabetic properties of the raw material were
interestingly described in a review paper by Drug. The results of the preclinical studies were
promising. Animal studies have shown its ability to lower blood glucose levels. Withaferin A can
effectively control induced type 1 diabetes in rats through modulation of Nrf2/NFκB signaling and
therefore has significant potential for therapy. In silico studies have also confirmed withaferin A’s
potential using molecular docking. However, only one clinical study from the year 2000 showed a
direct blood-glucose-lowering effect. On the other hand, many studies have shown a beneficial
effect on the lipidemic profile. In a study conducted on white albino rats with hypercholesterolemia,
a reduction in cholesterol levels and also in the antioxidant effects of Withania somnifera were
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

observed. In the case of clinical trials on diabetes, despite not showing an effect on blood sugar
levels, interesting results were achieved in improving the lipidemic profile, body weight, and blood
pressure, showed an improvement in the lipidemic profile and patient assessment with the DDS17
scale assessing patients’ distress scale. The administration of a standardized Ashwagandha extract
under the name SENSORIL improved antioxidant parameters and the lipidemic profile and
demonstrated the tolerability and safety of the raw material. In site of its tolerability and safety,
demonstrated an effect on lipidemic profile and a change in the reflection index. [21-22]

4.3. Scientific Name: Avena sativa L

Kingdom Plantae
Super division Spermatophyta
Division Magnoliophyta
Class Liliopsida
Order Cyperales
Family Poaceae
Genus Avena

Species A. Sativa
Common Names Oat, Cultivated Oat, Oats, Side Oat, Tree Oat, Red Oat

Figure 4. Avena sativa L

 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Description:
The word Avena originated from the Sanskrit word “avi,” which means sheep, or “avasa” means
food material. The Indian variety is equated with A. byzantina C. Koch., Avena sativa L. known as
oat or common oat is associate degree annual grass concerning 1.5 meters high, culms tufted or
solitary, erect or bent at the bottom. The leaves are non-articulate, green; the ligules are blunt and
membranous. The inflorescence may be a diffuse raceme with 2–3 florets, all bisexual or the distal
one or 2 could also be reduced and male or sterile; glumes sub-equal 7–11 patterned, longer glumes
17–30 mm; lemmas 7–9 patterned, either divided or with a mind their apex; lowest lemma is 12–
25 metric linear unit. The rachilla of the cultivated oat doesn't disjoint at maturity (that of many
weed species do). Its lemmas are seldom awed. The grain is tightly enveloped by the exhausting
lemma and palea. Seed size is found to vary with the variety. [23]

Pharmacological Activities
 Antidiabetic Activity
Foods yielding low postprandial glycemic responses have been suggested to be beneficial for the
dietary prevention of type 2 diabetes. The soluble fibre β-glucan from the oats can decrease the
glycemic response. The high molecular weight of β-glucan increases the fluid viscosity in the
intestine, it is crucial for obtaining the positive effect of β-glucan on the peak blood glucose level.
It has been reported that the inclusion of oat β-glucan into breakfast cereals could reduce the
postprandial glycemic response up to 50%. At low levels (below 5%) the response was dose
dependent whereas at the levels above 5% it did not show significant reductions in the glycemic
response. These findings will be of importance for deciding the appropriate levels of β-glucan
inclusion in food systems. Studies to evaluate the effect of inclusion of β-glucan enriched cereal in
foods was carried out in Type 2 diabetic patients and their glycemic indices (GI) were found to be
significantly lower than the GI of the group with white bread.

The results of the study suggest that the blood glucose levels of diabetic and pre-diabetic individuals
can be moderated by using β-glucan rich foods and so oat has great potential. Oats are a rich source
of magnesium which acts as a co-factor for more than 300 enzymes, including enzymes involved
in the body’s use of glucose and insulin secretion.

Foods containing at least 51% whole grains by weight (and are also low in fat, saturated fat, and
cholesterol) are permitted by FDA to display a health claim stating that its consumption is linked
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

to lower risk of heart disease and certain cancers. The data of an 8-year trial, involving 41,186
participants of the Black Women’s Health Study, revealed inverse associations between
magnesium, calcium, and major food sources in relation to type 2 diabetes Risk of type 2 diabetes
was 31% lower in the group of black women frequently eating whole grains as compared to those
eating the lesser quantity of those magnesium-rich foods. When the women’s dietary intake of
magnesium in take was considered by itself, a beneficial, but lesser-19%-reduction in risk of type
2 diabetes was found, indicating that whole grains offer special benefits in promoting healthy blood
sugar control. Daily consumption of low-fat dairy foods was also helpful in lowering the risk of
type 2 diabetes by 13%. [24]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

5. PLAN OF WORK
5.1. Comprehensive literature
5.2. Collection of crude drugs
5.3. Phytochemical screening
 Carbohydrates
 Alkaloids
 Tannins
 Glycosides
 Saponins
 Flavonoids
 Steroids (LB test)
 Proteins
 Amino acids
 Phenolic compounds
 Gums and Mucilage
5.4. Formulation of polyherbal antidiabetic cookies
5.5. Evaluation of physical properties of polyherbal antidiabetic cookies
 Phytochemical evaluation of powdered flower extract of Tecoma stans.
 Physiochemical properties of cookies.
 Nutritional analysis.
5.6. Results and discussion
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

6. MATERIALS AND METHODS

6.1. Materials
Table No 1. List of ingredients used
Name of ingredients Sources
Tecoma stans Local area, Chandrapur
Ashwagandha Local area, Chandrapur
Tulsi Local area, Chandrapur
Oats Local market, Chandrapur
Milk Local market, Chandrapur
Black gram flour Local market, Chandrapur
Flavoring Agents Local market, Chandrapur
Butter Local market, Chandrapur
Salt Local market, Chandrapur
Baking soda Local market, Chandrapur
Baking powder Local market, Chandrapur
Artificial sugar Diabliss Online Store, Chennai (TN)
Wheat flour Local market, Chandrapur

Table No 2. list of equipments used

Name of Equipment Make/Mode
Digital Weighing Balance MAB220, WENSAR
Soxhlet Apparatus LTSW-5, LAB TECH
Hot Air Oven S Clean-135947175, S Clean
Desiccator JII-1504, A SKY INSTRUMENT

 Collection of Sample
Fresh aerial parts of Tecoma stans (leaves), Ashwagandha (bark), and Tulsi (leaves) were collected
from local areas of Chandrapur city. The collected plants were washed properly and sun dried. The
dried plant was ground into coarse powder material and in sunlight container till future. Milk,
Flavoring agent (vanilla and cocoa), salt, baking powder, baking soda, butter, were purchased from
local market of Chandrapur and Artificial sugar (Diabliss sugar) was procured from its online store.
[25]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

6.2. Methods
 Preparation of Cookies
Different compositions of cookies were formulated using different ratios of Oats, wheat flour,
roasted black-gram flour, Tecoma stans leaves powder, Ashwagandha powder, Tulsi powder,
Milk, Flavoring agent (vanilla and cocoa), salt, baking powder, baking soda, butter, artificial sugar
(sugar free Natura), and based on the palatability and visual appealing final product were selected
for sensory evaluation and nutritional value analysis.

Figure 5. Preparation of cookie

 Preliminary batches for polyherbal cookies

Table No 3. Preliminary batches for polyherbal cookies
Content F1 F2 F3 F4 F5
Tecoma stans 15 gm. 13 gm. 15 gm. 12 gm. 20 gm.
Ashwagandha 10 gm. 8 gm. 12 gm. 10 gm. 5 gm.
Tulsi 5 gm. 9 gm. 3 gm. 8 gm. 5 gm.
Oats 40 gm. 40 gm. 40 gm. 40 gm. 40 gm.
Milk 10 ml. 10 ml. 10 ml. 10 ml. 10 ml.
Black gram flour 40 gm. 40 gm. 40 gm. 40 gm. 40 gm.
Flavoring Agents 1 ml. 1 ml. 1 ml. 1 ml. 1 ml.
Butter 15 ml. 15 ml. 15 ml. 15 ml. 15 ml.
Salt 1 gm. 1 gm. 1 gm. 1 gm. 1 gm.
Baking soda 1 gm. 1 gm. 1 gm. 1 gm. 1 gm.
Baking powder 1.5 gm 1.5 gm 1.5 gm 1.5 gm 1.5 gm
Artificial sugar 10 gm. 10 gm. 10 gm. 10 gm. 10 gm.
Wheat flour 100 gm. 100 gm. 100 gm. 100 gm. 100 gm.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

7. EVALUATION PARAMETERS
7.1. Phytochemical Evaluation of Powdered Flower Extract of Tecoma stans
The powdered flower sample of Tecoma stans was taken and phytochemical screening was done
to check the phytoconstituents present using standard reagents.

a) Carbohydrates
Molisch's test:
About 2ml of powdered flower extract was mixed with 0.2 ml of alcoholic solution of α- naphthol
10% in addition to 2 ml of sulphuric acid, a bluish violet zone is formed this indicates the presence
of carbohydrates and /or glycosides.

b) Alkaloids
About 0.2 g of the powdered flower extracts was warmed with 2% H2S04 for two minutes. It was
filtered and few drops of Dragendroff’s reagent were added. Orange red precipitate indicates the
presence of alkaloids.

c) Tannins
Small quantity of powdered flower extract was mixed with water and heated on water bath. The
mixture was filtered and ferric chloride was added to the filtrate. A dark green solution indicates
the presence of tannins.

d) Glycosides
The powdered flower extract was hydrolysed with HCl solution and neutralized with NaOH
solution. A few drops of Fehling’s solution A and B were added. Red precipitate indicates the
presence of glycosides.

e) Saponins
About 0.2 g of the powdered flower extract was shaken with 5ml of distilled water and then heated
to boil. Frothing (Appearance of creamy miss of small bubbles) shows the presence of saponins.

f) Flavonoids
Powdered flower extract of about 0.2 g was dissolved in diluted NaOH and HCl was added. A
yellow solution that turns colourless, indicates the presence of flavonoids.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

g) Steroids (LB test)

2 ml of acetic anhydride was added to 0.5 g of the powdered flower of each with 2 ml of H2 S04.The
colour changed from violet to blue or green in samples indicating the presence of steroids.

h) Proteins
To the powdered flower extract, 5%NaOH and 1% copper sulphate solution were added. Violet
color produced shows the presence of proteins.

i) Amino acids
The powdered flower was treated with Million’s reagent. Red colour showed the presence of amino
acids.

j) Phenolic compounds
Small quantities of powdered flower samples were taken separately in water and test for the
presence of phenolic compounds was carried out by using reagents like 5%ferric chloride solution,
1%gelatin solution containing 10% NaCl and 10%lead acetate.

k) Gums and Mucilage

A small quantity of powdered flower extracts were added separately to 25ml of absolute alcohol
with stirring and filtered. The precipitate was dried in air and examined for its swelling properties.
No swelling was observed indicates the absence of gums and mucilage. [8]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

7.2. Physiochemical Properties of Cookies

a) Moisture content:
Moisture content was determined by the method prescribed in the chemical Analysis of the
food. As stated in the procedure, sample of the cookies were weighed precisely in a moisture
dish and were kept in hot air oven for about 2 hours at 105°C and then it was cooled in
desiccators and again weighed. The process of heating was repeated for 30 min. and then again
cooled and weighed. The procedure was done until the variance between two successive
weighing became lessthan that of 0.001 gm. Moisture content in the test sample was calculated
which was based on the equation given below:

Where,
W1 = Weight of moisture dish with sample before drying;
W2 = Weight of moisture dish with sample after drying;
W = Weight of moisture dish.

b) Ash value:
Total ash content of the prepared cookies was determined by the following procedure.
According to the given procedure 1 gm of cookie sample was taken in a tarred crucible and it
was burnt on the Bunsen burner until all the carbon was burnt. Then Sample was let to be
cooled, weighed and then again the procedure was repeated until the weight became constant.
After that the total Ash value were calculated based on the equation given below:

Where,
W2 = Weight of empty dish;
W = Weight of sample taken;
W1=Weight of crucible with sample after complete burn.

c) Total alcoholic and water extractive values:

For the analysis of total alcohol/Water extractive value, 5 gm of cookies powder sample were
taken in 250 ml of volumetric flasks in which 90% ethyl alcohol or Distilled water were added
and was kept aside for 24 hours. After the completion of 24-hour samples were filtered and
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

were taken in porcelain dishes. All the samples of alcoholic and water extracts were heated at
the temperature of 100°C for evaporation, following samples were cooled down and advance
calculations were done by the following method.
Calculation: 5 gm of sample gives 4x of alcohol extract so 100 gm of sample gives = 80 × x/4
Where, x=Sample after drying. [2]

7.3. Nutritional analysis

a) Protein estimation:
Protein estimation was done by prescribed procedure in the given DGHS Manual. According
to this method 200-300 mg of cookies powder were taken in 4 test tubes and then 3 gm of
catalyst (K2SO4+CuSO4) was added in it. 10 ml of concentrated sulphuric acid H2SO4 was
added to all tubes and then digested for 3-4 hrs. Later these samples were distilled with boric
acid, potassium permanganate and 40% of Sodium hydroxide and then were titrated with acid.
This titrant was neutralized with ammonia and by this % of protein was calculated by using
following equation.

b) Fat content:
According to the given procedure, 2 gm of the cookie sample was kept in Soxhlet apparatus
with diethyl alcohol and petroleum ether in the ratio of 1:1 for 6 hours then ether was removed
by the process of distillation and were dried in hot air oven at 110 ± 1°C and later on was cooled
in a desiccator. Taken dried sample was weighed again. The left Residue was washed with 2
to 3 ml of diethyl ether and the same process was repeated until the weight became constant.

% of fat content = (M1-M2) x 100/weight of the sample

Where,
M1 =Weigh of Round bottom flask with fat;
M2 = Weigh of the Round bottom flask.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

c) Carbohydrate estimation:
Carbohydrate estimation was done by the procedure given in DGHS Manual. For estimation of
carbohydrate 2 gm of cookie sample powder was taken in a 200 ml of volumetric flask and
then 50 ml of lead acetate was added. 6 ml of 0.5 N HCl was added and heated on hot water
bath. After heating, the sample was cooled and neutralized with 6 ml of 0.5 N NaOH, lastly the
sample volume was makeup upto 200 ml using distilled water, Invert sugar was determined
before inversion by Lane and Eynon method.

According to this method 10 ml of mixed Fehling A and B solution was taken in the conical
flask and titration was passed out with sample solution within 3 min without inversion by using
1% aqueous Methylene Blue as an indicator.

Reducing sugar % before inversion =F X 10/C X R

Where, C = concentration; R = Reading; F = Factor of Fehling solution
Total invert sugar % after inversion =F X 10/C X R
C = concentration; R = Reading; F = Factor of Fehling solution
Total carbohydrate = total invert sugar after inversion – invert sugar % before inversion x 0.95

d) Total Energy
Total energy was valued on the basis of carbohydrates, proteins and fats content of cookie
sample.
Energy (Kcal) = Fat×4+ Protein×9+ Carbohydrates×4

e) Sensory Analysis
Sensory attributes such as flavor, aroma, taste, appearance and odor were evaluated by 9- point
hedonic scale, total 64 people participated in this survey. Questionnaires and mouth rinsing
water were conferred to taste panelist, through the session product was introduced and
questions were explained to the volunteers. The obtained data were analyzed by Microsoft
Excel on the basis of age group. [27]
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

8. RESULTS AND DISCUSSION

Table 4: Chemical and physiochemical parameter

Sr. no Chemical and physiochemical Results
parameters
1 Ash content 7.10%
2 Moisture content 6.91%
3 Alkaloid Present
4 Alcohol extraction 6.58%
5 Water extraction 5.30%
6 Fat content 14.04%
7 Carbohydrate content 60.51%
8 Protein content 11.65%
9 Total energy 414.9874 Kcal

By above mentioned Table 1 of results shown, those Cookies were spanking in nutritional values
with controlled fat and carbohydrate and high rich in protein content which have made it
acceptable among the health-conscious people, growing people and case of malnutrition.
Comparative studies of different parameters of sensory evaluation exhibited that use of curry
leaves as an active ingredient has given an appetizing and flavoring effect. Selected composition
of cookies has made it acceptable around 90-96% that will give us a hope to convert this
formulation into large scale production. In terms of physiochemical properties, nutrition value,
sensory evaluation and comparison with other marketed product were in acceptable range.

8.1. Sensory Evaluation

Taste is a strategic parameter of sensory evaluation. The product might be captivating and having
an eminent energy but sans of righteous taste it is likely to unaccepted. So, on the basis of sensory
evaluation, it is found that due to use of Tecoma stans leaves its mean score was 90-95% among
the age group of 18-40. Graph of taste has been depicted below (Figure No. 6).
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

Figure 6: Acceptance of taste

8.2. Flavour
Flavour is an integral part of taste that plays a vital role in the acceptance of any food material.
Used flavour of Tecoma stans leaves and coco was found to be highly appreciable among all the
volunteer of sensory evaluation because its smell act as an appetizer, because of that it means score
on excel was found to be 90-95% (Figure No. 7).

Figure 7: Acceptance of flavor

8.3. Aroma and Color
Aroma or fragrance of foods are the indiscernible part of the acceptance and play a crucial role in
mouth feel. Aroma is the first sign of consumer to choose any food for consumption as well as
color of food product is a sign of acceptance that have a great impact on the choice of any product.
Aroma and color show an elegant effect on the acceptance.
The outcome of results showed that strong combined Tecoma stans leaves, Ashwagandha and Tulsi
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

powder’s aroma influenced the people greatly.

8.4. Overall Acceptance of Cookies

It is very important for food, snacks, and soft drinks that after taking the bite or ship it must give a
palatable and flavourful effect on tongue so it could be taken easily. The results of sensory
evaluation on the basis of taste, flavour, crispiness and aroma were found to be appreciable with
medicated effect.

8.5. Comparative Study of Nutritional Factor

Nutritional factor is the main aim of this study. After completion of all evaluation when it was
compared with different brands of biscuits and it was found that it has less fat and carbohydrate
which is highly beneficial for diabetic patient as well as high protein content due to use of black
gram powder, this make it protein rich which might be use full for growing and malnourished
children (Figure No. 8).

Figure 8: Comparative studies with other marked product

 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

9. CONCLUSION
Based on statistical data it was found that herbal cookies have high amount of protein, controlled
amount of fat and carbohydrate in compared to other marketed product. Presence of Tecoma stans
leaves and ashwagandha bark have given a new direction to convert home remedies into a marketed
preparation which gives the highest acceptance in terms of flavor as well as the ingredients used in
formulation has already proven significant role to control non-insulin dependent diabetes mellitus
and other health issue. So, it could be serviceable in the manufacturing of highly nutritious cookies
with low cost and high efficiency.
 Formulation And Evaluation of Polyherbal Cookies For Diabetic Patient

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