Journal of Pharmacognosy and Phytochemistry 2019; 8(4): 3529-3535

E-ISSN: 2278-4136
P-ISSN: 2349-8234
JPP 2019; 8(4): 3529-3535 Formulation and evaluation of immune boosting
Received: 06-05-2019
Accepted: 10-06-2019 herbal tea
Sushmita L Bhandare
Department of Pharmacognosy, Sushmita L Bhandare and Smita P Borkar
Arvind Gavali College of
Pharmacy, Jaitapur, Satara,
Abstract
Shivaji University Kolhapur,
The evaluation of anti-inflammatory activity was carried out on the crude aqueous extract of the flowers
Maharashtra, India
of Moringa oleifera. The aim of this study was to evaluate the anti-inflammatory activity of Moringa
Smita P Borkar oleifera flowers against Diclofenac Sodium, also to evaluate the Anti-inflammatory activity of prepared
Department of Pharmaceutics, Herbal Tea formulation against Diclofenac Sodium as an standared. The anti-inflammatory activity of
Arvind Gavali College of aqueous flower extract was determined in vitro, by inhibition of thermally induced protein denaturation.
Pharmacy, Jaitapur, Satara, The Moringa oleifera flower extract showed significant inhibition of denaturation of egg albumin in dose
Shivaji University Kolhapur, dependent manner. This result provide valuable information that Moringa oleifera hold great promise as
Maharashtra, India highly effective as an anti-inflammatory agent.

Keywords: Moringa oleifera, anti-inflammatory activity, pharmacological activities, diclofenac sodium

Introduction
The sensory appeal of tea, like a all food products, is an important consideration in new
product development. Tea in general and herb tea in particular are gaining increasing
consumer attention due to growing awareness of health benefits derived from their
consumption. Even though several under utilized plants exits with potential for processing into
herb tea, research in product development of herb tea is limited [1].
The herbal teas are made from herbs, fruits, seeds, roots steeped in hot water. Instant tea may
contain very little amounts of actual tea and plenty of sugars. A pharmaceutical branch of
Ayurveda has contributed several innovative dosage forms. Conversion of dosage form into
more suitable for modern era with additional benefits of palatability and presentation is always
essential [1].

Need of present investigation

 Research work on same plant was done by various ways, in current investigation we will
the study about flowers of the plant.
 Formulation of investigation plant is unique and easy to use and shall take regular as a
health drink.

Fig 1: Drumstick tree

Aim
Correspondence “Formulation and evaluation of immune boosting herbal tea”.
Sushmita L Bhandare
Department of Pharmacognosy, Objectives
Arvind Gavali College of  To prepare immune boosting herbal tea.
Pharmacy, Jaitapur, Satara,
Shivaji University Kolhapur,
 To evaluate the formulation with respect to various physical parameter.
Maharashtra, India
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 To evaluate the phytochemical screening of given crude  To evaluate the formulation with respect to anti-
drug. inflammatory activity.

Table 1: Plan of work

S. No Title
1 Literature review.
2 Selection of drug.
3 Crude drug profile.
4 Materials and method.
5 Experimental work.
6 Formulation of herbal tea.
7 Evaluation of herbal tea.
8 Result and conclusion.
9 Reference.

Literature review

Table 2: List of reference papers

S. No. Title of paper Author Information
Isolation and characterization of coagulant
Tetsuji Okuda; Aloysius U. It discovered coagulant extract from Moringa olifera seed
1. extracted form of moringa olefera seed and
Baes by salt solution.
salt solution.
Effect of Moringa oleifera leaves aqueous Dolly Jaiswal, Prashant
2. It estimate the study of hypoglycemic agents on rats.
extract therapy on hypoglycemic rats kumar Rani, Amit kumar
Effect of antioxidant activity of Moringa It discovered that antioxidant provide protection against
3. Shahid Iqbal, M.I. Bhanger
oleifera leaves degenerative diseases including cancer, Alzheimer disease.
Evaluation of Aqueous leaves extract of It includes aqueous extract of leaves of Moringa oleifera
B.S. Rathi, S.L. Bodhankar
4. Moringa oleifera for wound healing in was investigated and rationalized for its wound healing
and A.M. Baheti
albino rats activity.
Pharmacological studies on Hypotensive and Anwar H. gilani, Khalid It estimate the study of hypotensive, spasmolytic activity
5.
spasmolytic activities of Moringa oleifera Aftab exhibited by Moringa oleifera constituent.
The review work deals with nutritional, Khawaja Tahir Mahmood,
Nutritional values of Moringa oleifera compared with other
6. therapeutic, traditional uses or benefits of Tahir Mugal and Ikram UL
food.
moringa. Haq.
Potential uses of Moringa oleifera and
Reminder that heroic lengths and modern science are not
examination of antibiotic efficacy conferred Rockwood J.L, Anderson
7. always necessary to combat antimicrobial pathogens in
by M. oleifera seed and leaf extract used by B.G, Casamatta D.A.
remote regions where modern medicine are not available.
crude extraction techniques.
Bhoomika R Goyal, Babita B
Phyto-pharmacology of Moringa oleifera M. oleifera mainly contains alkaloids, flavonoids,
8. Agrawal, Ramesh K Goyal
Lam, An overview. anthocyanins, proanthocyanidins and cinnamates.
and Anita A Mehta.

Selection of drug dipropionate and progrsterone and shown incresed

Moringa oleifera is one of the miracle tree, is widely histoarchitecture of uterine [8].
cultivated throughout India, belong to Family Moringaceae. It
is widely used as a nutritive herb and possess valuable 3. Anti-oxidant acitivity
pharmacological activities. present article describes habitat, Moringa oleifera exhibit strong anti-oxidant and radical
pharmacogonstic features hytochemistry, nutritive values and scavenging activity [8].
pharmacological activities like anticancer, antimicrobial, anti-
inflamatory, antihyperlipidemic, hypotensive, antidiabetic, 4. Cardiovascular activity
hepatoprotective, antiasthemetic, anthelminic, anti-fertility, Ethanolic extract of Moringa oleifera shows antihypertensiv
etc of moringa. It is one of the rich sources of vitamin C, milk or hypotensive activity. It was found that thiocarbamate and
protein, etc. Present review gives the information of all isothiocyanate glycosides are responsible for this promising
essential nutrients that are needed to improve immunity [8]. hypotensive activity [8].

5. Anti-epileptic activity
Pharmacological activities
Methanolic extract of Moringa oleifera were investigated its
1. Anti-cancer activity
anti-convulsant activity using pentylenetetrazole (PTZ) and
Various extracts of leaves and ethanolic extract of seeds of
maximum electric shock (MES) on male albino mice [8].
Moringa oleifera shows anti tumor activity in-vitro tests.
Thiocarbamate and isothiocyanate related compounds were
6. Anti-asthmatic activity
isolated, which act as inhibitor of tumor promoter teleocidin Moringa oleifera were found spasmolytic in Acetylcholin,
B-4-induced Epstein Barr virus (EBV) activation in Raji cells [8]. histamine, Bacl2 and 5HT induced bronchospasm [8].
2. Anti-fertiliy activity 7. Anti-ulcer activity
Aqueous extract of Moringa oleifera was found be effective Antiulcer activity in various animal models on adult holtzman
as an anti-fertility in presence as well as absence of estradiol albino rats of either sex [8].
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8. Anti-inflammatory activities a) Indoleacetic acid: Indomethacin

Methanolic extract of leaves and flowers as well as ethanolic b) Indeneacetic acid: Sulindac
extract of seeds of Moringa oleifera has shown anti- c) Pyrroleacetic acid: Tolmentin,
inflammatory activity in carrageenan induced paw edema d) Phenylacetic acid: Ibuprofen, Diclofenac
model. Aurantiamide acetate and 1, 3 dibenzyl urea, isolated
from roots shown this intiinflammatory activity so they 6. Oxicams-
responsible for anti inflammatory activity of Moringa Piroxicam
oleifera. anti-inflammatory agents are used to cure
inflammation caused by prostaglandin (PGE2). Drugs with 7. Miscellaneous
analgesic, antipyretic, and anti-inflammatory effects -they Nimesulide
reduce pain, fever, inflammation [8].
MOA
Classification Inhibitors of the enzyme cyclo-oxygenase, inhibiting both the
1. Salicylic acid derivatives- sodium salicylate, Aspirin cyclo-oxygenase-1 (COX-1) and cyclo-oxygenase-2 (COX-2)
2. p-Aminoplenol derivatives - paracetamol, phenacetin isoenzymes. Cyclo-oxygenase catalyses the formation of
3. Pyrazolidinedione derivatives- Plenylbutazone prostaglandins, prostacyclin and thromboxane from
4. Anthranilic acid derivatives - Mefenamic acid, arachidonic acid (Which is derived from the cellular
Meclofenamate phospolipid bilayer by phospolipase A2). Prostaglandins act
5. Aryl alkanoic acid derivatives- as messenger molecules in the process of inflammation.

Fig 2: Mode of action of Anti-inflammatory drugs

Crud drug profile Marathi: Shevga

Introduction to Moringa oleifera Hindi: Saguna, sainjna
Name: Moringa oleifera English: Drumstick tree
Tamil: Morigkai
Synonyms [8] Family: Moringaceae
Sanskrit: Subhanjan

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borneol (0.5%), zingiberene (25%) and sesquiterpines (53%).

Curcumin (diferuloylmethane) (3–4%) is responsible for the
yellow colour, and comprises curcumin I (94%), curcumin II
(6%) and curcumin III (0.3%) Curcumin was first isolated in
1815 and its chemical structure was determined by Roughley
and Whiting 9 in 1973 melting. It has a point at 176–177 °C;
forms a reddish-brown salt with alkali and is soluble in
ethanol, alkali, ketone, acetic acid and chloroform [4].

Pharmacological actions of curcumin

1. Effect on cardiovascular system
Curcumin decreases severity of pathological changes thus
protects from damage caused by myocardial infarction.
Curcumin improves Ca2+ -transport and its slippage from the
cardiac muscle sarcoplasmic reticulum, thereby raising the
Fig 3: M. oleifera flower
possibility of pharmacological interventions to correct the
defective Ca2+ homeostasis in the cardiac muscle. Curcumin
Taxonomic classification [8]
has significant hypocholesteremic effect in
Kingdom: Plantae
hypercholesteremic rats [4].
Sub kingdom: Tracheobionata
Super division: Spermatophyta 2. Effect on nervous system
Division: Magnoliopsida Curcumin and manganese complex of curcumin offer
Class: Magnoliopsida protective action against vascular dementia by exerting
Sub class: Dilleniidae antioxidant activity [4].
Order: Capparales
Genus: Moringa 3. Anti-inflammatory activity
Species: oleifera Curcumin is effective against carrageenin-induced oedema in
rats and mice. The natural analogues of curcumin, viz. FHM
Termeric and BHM, are also potent anti-inflammatory agents. The
Synonyms [4]: Curcuma longa, Curcuma herb volatile oil and also the petroleum ether, alcohol and water
extracts of Curcuma longa show anti-inflammatory effects [4].

4. Anticoagulant activity
Curcumin shows anticoagulant activity by inhibiting collagen
and adrenaline-induced platelet aggregation in vitro as well as
in vivo in rat thoracic aorta [4].

5. Antidiabetic effect
Curcumin prevents galactose-induced cataract formation at
very low doses. Both turmeric and curcumin decrease blood
sugar level in alloxan-induced diabetes in rat. Curcumin also
decreases advanced glycation end products induced
complications in diabetes mellitus [4].

6. Antifungal effect
Ether and chloroform extracts and oil of C. longa have
Fig 4: Termeric antifungal effects. Crude ethanol extract also possesses
antifungal activity. Turmeric oil is also active against
Biological Source: It is dried root and rhizomes of Curcuma
Aspergillus flavus, A. parasiticus, Fusarium moniliforme and
longa.
Penicillium digitatum [4].
Family: Zingiberaceae
7. Antivenom effect
Taxonomical Classification [4] Ar-turmerone, isolated from Curcuma longa, neutralizes both
Kingdom: Plantae. haemorrhagic activity of Bothrops venom and 70% lethal
Division: Magnoliophyta
effect of Crotalus venom in mice. It acts as an enzymatic
Class: Liliopsida
inhibitor of venom enzymes with proteolytic activities [4].
Subclass: Commelinids
Order: Zingiberales Cardamom
Genus: Curcuma Species Curcuma longa
Synonym [5]: Choti – Ilalchi (Hindi)
The wild turmeric is called C. aromatica and the domestic
Biological source: Ripe fruit of Elettaria cardamomum var
species is called C. longa.
Family: Zingiberacea
Chemical composition
Turmeric contains protein (6.3%), fat (5.1%), minerals Taxonomic classification [5]
(3.5%), carbohydrates (69.4%) and moisture (13.1%). The Kingdom: Plantae
essential oil (5.8%) obtained by steam distillation of rhizomes Order: Zingiberales
has a-phellandrene (1%), sabinene (0.6%), cineol (1%), Genera: Elettarai Amomum
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Equipments

Table 4: List of equipments used

S. No. Instrument Make & Model
1. Incubator Quality, QBOD-05
2. Hot air oven Bio-Techniqcs India, BTI-26
3. Vaccum pump Value, VE115N
4. Refrigerator Blue Star, CHF150C
5. Electronic balance Shimadzu, BL-22OH
6. Uv Spectrophotometer Dynamic, Halo DB-20
7. Muffle Furnance Hally Instruments, HI-25

Fig 5: Cardamom seeds Experimental work

Preparation of herbal tea [7]
Chemical composition
Seeds of Elettaria cardamomum are rich in volatile oil that General method of preparation emphasized for shatavari
mainly includes phenolic and flavonoid components. Starch, granules is follow preparation of Moringa oleifera herbal tea.
protein, waxes and Sterols are other components of the o. Coarse powder of Moringa oleifera flowers

Pharmacological activities Mixed with sugar syrup

1. Antibacterial activity The mixture was heated on mild fire (Mandagni) i.e 90 0C-
Ethanolic extract of E. cardamomum possess antibacterial 1000c till it attained more than two thread consistency of
effect at the dose of 512 μg/mL. Toxicity of the extract was sugar syrup
observed at 0.3 mg/g, which showed inflammation in brain,  1 hrs 30 min of heating –adhesion of syrup to spoon.
oxidative stress and cells necrosis in heart. The use of E.  1 hrs 50 min of heating-syrup was found to be in a two
cardamomum as spice should not exceed the 0.003 mg/g since thread consistency.
at this amount no negative effects were observed [5].  2 hrs 5 min of heating-not instant dissolution in water.

2. Gastroprotective activity Add Turmeric as a Anti-inflammatory agent and also as a

Gastroprotective activity of E. cardamomum was best found colouring agent
in the petroleum ether soluble extract which inhibited lesions
by nearly 100% at 12.5 mg/kg in the aspirin-induced gastric Add cardamom as a flavouring agent
ulcer. Methanolic extract also possess gastro protective effect
[5]
. The contents were removed from heat source

3. Antioxidant activity Thus obtained mass was dried in hot air oven and subjected to
Cardamom oil is effective as an antioxidant and can increase multi mill sieve to obtain granules
levels of glutathione, a natural antioxidant in body. The effect
is increased by increasing the content of the oil from 100 to Herbal tea
5000 ppm [5].
Formulation of herbal tea
4. Insecticidal activity
The volatile oil from cardamom acts as a potential grain Table 5: Formula for herbal tea
protectant by killing various life stages of the stored product
Formulation
insects attacking wheat, e.g. Tribolium castaneum and Ingredients
F1(gm) F2(gm)
Sitophilus zeamais, via contact and fumigant action [5]. Moringa oleifera Flowers 2 8.33
Sugar 10 41.66
Materials and Methods Turmeric 0.030 0.125
Materials Cardamom 0.030 0.125
 Moringa oleifera flowers.
 Turmeric. Extraction of plant material
 Cardamom. In the present study, the dried powdered flower were
extracted by aqueous extraction method at room temperature
Methods with occasional shaking. The extract was filtered and re-
To determine anti-inflammatory activity of Moringa oleifera extracted by same process until plant material were
flowers. exhausted. The collected filtrates were pooled and evaporated
to dryness under reduce the pressure to yield dry extract and
Chemicals was stored at 40c until used.

Table 3: List of chemicals used Result and Discussion

S. No Name of ingredients Source Evaluation of in-vitro anti-inflammatory activity
Diclofenac Sodium Acme Generics LLP Davni, Tehsil Anti-inflammatory activity of M. oleifera flower extract was
1. evaluated by protein denaturation method. Diclofenac
tablet IP 50mg Nalegarh.
Phosphate Buffer Saline Sodium, a powerful non -steroidal anti-inflammatory drug
2. A.G.C.O.P. Satara
pH 6.4 was used as standard drug.
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Reaction mixture consists of 2 ml of different concentrations % water soluble ash value =

of M. oleifera flower extract (ug/ml) or standared Diclofenac
Sodium (ug/ml) and 2.8 ml of Phosphate Buffered Saline pH Wt. of total ash-Wt of water insoluble ash × 100
(6.4) was mixed with 0.2 ml of egg albumin (From fresh Wt. Of crude drug taken
hen’s egg) and Incubated at (27 0C) for 15min. Denaturation
was induced by keeping the reaction mixture at 70 0C in a = 2.85 %w/w
water bath for 10 min. After cooling, the absorbance was
measured at 660 nm by using double distilled water as blank. 3. Acid insoluble ash value
Each experiment was done in triplicate and average was Boiled the ash for 5 min with 25 ml of 2 M HCL. Filtered and
taken. The percentage inhibition of protein denaturation was collected the insoluble matter on ash less filter paper, washed
calculated by using following formula [6]. with hot water and ignited in tarred crucible the temperature
not exceeding 450 0C for 4 hrs. Cooled in dissicator and
% inhibition = At –Ac ×100 weighted. Calculated of percentage of acid insoluble ash with
Ac the reference to the air dried drug.

Were, Wt. of acid insoluble ash

% Acid insoluble ash value = × 100
At =Absorbance of Test Wt. of crude drug taken
Ac=Absorbance of Standard/control
=3.45 %w/w
Evaluation parameters
Determination of ash value 4. Bulk density (gm/ml)2
Ash value are helpful in determining the quality and purity of  Bulk volume in ml
crude drug, especially in powder form. The objective of ash  Mass of Granule in gram
vegetable drugs is to remove all traces of organic matter,
which may otherwise interfere in an analytical determination. Bulk density = Mass
Bulk Volume =0.60 g/ml
1. Total Ash value
Weight accurately about 2 gm of powdered drug in a tarred 5. Tapped density (gm/ml)2
silica crucible. Incinerated at a temperature not exceeding 450  Tapped volume in ml
0
C for 4hrs, until free from carbon, cooled and wighted.  Mass of granule in gram

Mass
Wt. Of total ash Tapped density =
% Total Ash value = × 100 Tapped Volume
Wt. Of crude drug taken
= 0.78 g/ml
= 7.9 %w/w
6. Angle of repose2
2. Water soluble ash value  Height of pile in cm
The ash boiled with 25 ml of water, filtered and collected the  Average radius of circle in cm
insoluble matter on an ash less filter paper, washed with hot
water and ignited in a tarred crucible at temperature not Angle of repose (ɵ) = tan-1(h/r)
exceeding 450 0C for 4 hrs cooled in desiccators, weighted =18.26
and sub stracted wight off insoluble matter from the total
weight of ash. Preliminary phytochemical analysis of M. oleifera flower
extract [3]

Table 6: Preliminary phytochemical screening of M. oleifera flower extract

Phytochemical tests Name of test Flower extract
Tannins 5%FeCL3,Lead acetate test +
Steroids Salkowski test +
Flavonoids Shinoda test +
Hagers test
Alkaloids Meyers test +
Wagners test
Carbohydrates Molishs test +
Terpenoids Salkowski test +
Cardic Glycoside Keller killiani test +

In vitro Anti-inflammatory effect of M. oleifera flower extract

Table 7: % inhibition of protein denaturation by M. oleifera flower extract

Conc.(ug/ml) Absorbance of test (Extract) Absorbance of reference (Diclofenac sodium) % Inhibition of protein denaturation
100 0.0837 0.0489 71.16
400 0.1553 0.0621 150.08

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In vitro Anti-inflammatory effect of prepared herbal tea formulation

Table 8: % inhibition of protein denaturation by prepared herbal tea formulation

Conc.(ug/ml) Absorbnce of test (formulation) Absorbance of reference (Diclofenac sodium) % inhibition of protein denaturation
100 0.0927 0.0489 89.57
400 0.1270 0.0621 104.50

Conclusion
The Medicinal Plants since ancient time are lauded for their
diverse pharmacological actions which could be attributed to
presence of secondary plant metabolites such as alkaloids,
flavonoids, glyacosides, tannin etc.
The result of our study suggest that the aqueous extract of
Moringa oleifera flowers shows marked in-vitro anti-
inflammatory activity in dose dependent manner.

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