Studies on Phytochemical Constituents of Medicinal Plants
CHINTAN.PPT
1. 1
DESIGN DEVELOPMENT AND EVALUATION
OF NOVEL POLYHERBAL ANTIDIABETIC
TABLET FORMULATION
Pharmacy Department,
Faculty of Technology & Engineering
The Maharaja Sayajirao University of Baroda
PRESENTED BY:
Chintan Chauhan
GUIDED BY:
Prof. S. H. Mishra
2. OVERVIEW
2
1) Introduction
2) Herb that used traditionally in diabetes
3) Literature review
4) Objective
5) Experimental design
6) Procurement of raw material
7) Evaluation of raw material
8) Phytochemical studies of raw material
9) Thin layer chromatography(TLC) of Each extract
10) Selection of Dose for formulation
11) Anti hyperglycemic activity of each combination
12) Future prospects
4. 5
Plant Name Botanical name
and family
Part use Chemical
constituents
Biological use
Gudmar Gymnema
sylvestre,Asclepediaceae
Leaves Gymnemosides,gymn
emic acid(I-IV),resin
stigmasterol etc
Anti-Diabetic agent
Bitter
gourd(Karela)
Momordica
charantia,Cucurbitaceae
fruit Momordicin,charanti
n,ascorbic acid
Anti-Diabetic agent
Indian
Kino(Vijaysar/Bij
asar)
Pterocarpus
marsupium,Fabaceae
Bark, heartwood Pteroside
,epicatechin,sitosterol
,lupeol
Anti-Diabetic Agent
Jamun Syzygium cumini
(Eugenia
jambolana),Myrtaceae
Seed,fruit,leaves,k
ernel
Anti-arthritic,Anti-
Diabetes,Gastric
Ulceration
Ginger Zingeber
officnale,Zingeberaceae
Rhizome Gingerol,Shagoal,sesq
uiterpene
Anti-
Arthritic,Antidaibetes,car
diovascular disease
Outline of plants selected for further studies
5. Literature review
6
Gymnema sylvestre:
Antidiaretic effect of a leaf extract from gymnema sylvestre in non-insulin-dependent diabetes mellitus
patients, K. BASKARAN, Journal of Ethno pharmacology, 30 (1990) 295 – 305
In vitro callus and in vivo leaf extract of Gymnema sylvestre stimulate –cells regeneration and anti-
diabetic activity in Wistar rats, A. Bakrudeen Ali Ahmeda, journal of Phytomedicine,
Momordica charantia:
A medicinal potency of momordica charantia, D. Sathish Kumar, International Journal of
Pharmaceutical Sciences Review and Research.
Pharmacological actions and potential uses of Momordica charantia: a review, J.K. Grover, Journal
of Ethno pharmacology 93 (2004) 123–132
6. 7
Eugenia jambolana
Anti-diabetic activity of Syzygium cumini and its isolated compound against streptozotocin-induced diabetic
rats, A. Kumar, Journal of Medicinal Plants Research Vol. 2(9), pp. 246-249
Hypoglycemic and hypolipidemic effects of flavonoid rich extract from Eugenia jambolana seeds on
streptozotocin induced diabetic rats, Bhavna Sharma, Food and Chemical Toxicology, 46 (2008)
2376–2383
Pterocarpus marsupium:
Evaluation of anti-hyperglycemic and hypoglycemic effect of Trigonella foenum-graecum Linn,
Ocimum sanctum Linn and Pterocarpus marsupium Linn in normal and alloxanized diabetic rats, V.
Vats, Journal of Ethnopharmacology 79 (2002) 95–100
Zingeber officinale:
Effect of Ginger Extract Consumption on levels of blood Glucose, Lipid Profile
and Kidney Functions in Alloxan Induced-Diabetic Rats., Abd-Elraheem A. Elshater, Egypt.
Acad. J. biolog. Sci., 153-162 (2009)
9. Literature review
10
Plant Phytochemical Reference Biological Reference
Gymnema sylvestre Amino acids V. A. Khramov et al Stimulate β-cells
regeneration
A. Bakrudeen Ali Ahmeda
et al
dihydroxy gymnemic
triacetate
Pitchai Daisya et al
Regeneration of Islets of
langerhans
E.R.B.
shanmugasundaram et al
Triterpenoid saponins Niranjan p. Sahu et al
Momordica charantia Cucurbitane-type
triterpenoids(charantin)
Sook Young Lee et al Pharmacological Review J.K. Grover et al
Medicinal potency of
momordica charantia
D. Sathish Kumar et alMansoor ahmed et alTriterpenes, a sterol and a
monocyclic alcohol
Pterocarpus marsupium pterostilbene Ajanta Chakraborty et al Hypoglycemic activity of
Red kino tree
T.Radhika et al
aurone glycosides Achyut Narayan Kesariet
al
Upregulation of Glut-4
and PPAR
R. Anandharajana et al
Pterocarposide S. S. Handa et al Efficacy of P.Marsupium
in newly diagnosed
patients
ICMR group
Eugenia jambolana mycaminose A. Kumar et al effects of flavonoid rich
extract
Bhavna Sharma et al
Betulinic acid and 3,5,7,4
–tetrahydroxy flavanone
K.Karthic et al
Kasiappan Ravi et alantioxidant defense
system in STZ induced
diabetes
Zingeber officinale Volatile oils, gingerols,
shogaols
Natural Remedies Consumption on levels of
blood Glucose, Lipid
Profile
Abd-Elraheem et al
10. OBJECTIVE
11
Evaluation of selected herbal extracts according to WHO
guidelines and modern parameters
Optimization of different combinations of selected
extracts by Oral Glucose Tolerance Test in Rats
Optimization of excipients in selected combination used
in the formulation
Development and evaluation of herbal tablet
Phytochemical analysis of developed herbal tablet by
HPTLC
Anti-diabetic evaluation of optimized combination and
formulation of extracts by STZ-NAD induced
experimental model in rats
11. EXPERIMENTAL DESIGN
12
Procurement of Raw material.
1) Evaluation of Raw material
2) Phytochemical studies of each extract
Optimization of Combination:
1) Acute toxicity study as per OECD guidelines
2) Oral Glucose Tolerance Test in normal Rats
Development of dosage form(tablet)
1) Method of preparation
2) Selection of excipients
3) Optimization of excipient using factorial design
4) Evaluation of the dosage form
5) Stability study
Quality Assurance
1) HPTLC of the final dosage form
2) Total tannins evaluation
3) Total alkaloids evaluation
4) Total Flavanoids evaluation
5) Proximate analysis
Anti Diabetic Study
1) STZ-NAD induced Anti-Diabetic Study
12. PROCUREMENT OF RAW MATERIAL
Herbal extracts were obtained from AMSAR INDUSTRIES
LTD,INDORE. As a gift sample.
TYPE OF EXTRACT PART USED SOLVENT USED FOR
EXTRACTION
GYMNEMA SYLVESTRE DRY
EXTRACT
LEAVES HYDRO-ALCOHOLIC
MOMORDICA CHARANTIA DRY
EXTRACT
FRUITS AQUEOUS
PTEROCARPUS MARSUPIUM DRY
EXTRACT
BARKS ALCOHOLIC
EUGENIA JAMBOLANA DRY
EXTRACT
SEEDS AQUEOUS
ZINGEBER OFFCINALE DRY
EXTRACT
RHIZOME AQUEOUS
13
18. 19
Plant Extract Reported Acute toxicity
study (LD50)
Reference
Gymnema extract 3990 mg/kg in rats http://www.mdidea.com/pr
oducts/new/new020.html
Momordica extract 1200 mg/kg in rats Abalaka, M. E et al.
Pterocarpus extract >2000 mg/kg in rats T.Radhika et al.
Eugenia extract >2000 mg/kg in rats A. Kumar et al.
Ginger extract >2500mg/kg in rats Natural remedies document
SELECTION OF DOSE FOR FORMULATION
Acute toxicity study of each plant had been reported and they are as under.
19. 20
Acute oral toxicity study was performed as per OECD-423 guidelines (acute toxic
class method).Rats of either sex selected by random sampling technique were used
for the study. The animals were kept fasting for overnight providing only water,
after which the combination of extracts at a ratio of (1:1:1:1:1) was administered
orally at the dose level of 100 mg/kg body weight by intragastric tube and
observed for 14 days. If mortality was observed in 2 - 3 animals, then the dose
administered was assigned as toxic dose. If mortality was observed in one animal,
then the same dose was repeated again to confirm the toxic dose. If mortality was
not observed, the procedure was repeated for further higher dose 500, 1000 and
2000 mg/kg body weight.
This study showed no mortality up to the dose of 2,000 mg/kg body weight. So,
the combination of extracts is safe for long term administration.
Sr. No. Dose of
Combination
Mortality of
Rats
1 100 mg/kg 0
2 500 mg/kg 0
3 1000 mg/kg 0
4 2000 mg/kg 0
20. 21
Combination 1 Combination 2 Combination 3 Combination 4
Gymnema sylvestre 2
Momordica charantia 1
Pterocarpus marsupium 1
Eugenia jambolana 1
Zingeber officinale
1
Gymnema sylvestre 1
Momordica charantia 2
Pterocarpus marsupium 1
Eugenia jambolana 1
Zingeber officinale
1
Gymnema sylvestre 1
Momordica charantia 1
Pterocarpus marsupium 2
Eugenia jambolana 1
Zingeber officinale
1
Gymnema sylvestre 1
Momordica charantia 1
Pterocarpus marsupium 1
Eugenia jambolana 2
Zingeber officinale
1
Combination 5 Combination 6 Combination 7 Combination 8
Gymnema sylvestre 2
Momordica charantia 2
Pterocarpus marsupium 1
Eugenia jambolana 1
Zingeber officinale
1
Gymnema sylvestre 2
Momordica charantia 1
Pterocarpus marsupium 2
Eugenia jambolana 1
Zingeber officinale
1
Gymnema sylvestre 2
Momordica charantia 1
Pterocarpus marsupium 1
Eugenia jambolana 2
Zingeber officinale
1
Gymnema sylvestre 1
Momordica charantia 1
Pterocarpus marsupium 1
Eugenia jambolana 1
Zingeber officinale
1
Part
The total dose of the formulation was selected as 400 mg/kg in human and Dose for each extract
in combination was selected on random trial bases that were divided into parts.
21. Preliminary Screening of combination Through Oral Glucose
Tolerance Test.
22
Above selected combination were screened for oral glucose tolerance test in
normal rat for its anti-hyperglycemic activity.
Animals
Healthy female Sprague dawley rats weighing 250-350gm, procured from
Zydus Cadilla Research Laboratory, Ahmedabad maintained under standard
husbandry conditions (Temperature 23 ± 2 °C, relative humidity 55 ± 10%
and 12 hrs light dark cycle).
The animals were fed with standard rat pellet diet and had free access to
water. The experimental protocols were approved by the Institutional Animal
Ethics Committee, The M.S.University of Baroda, Vadodara, Gujarat.
22. 23
Preparation of solutions for Administration:
I Vehicle: 0.5 % CMC suspension was used as a vehicle.
II Metformin suspension: Standard metformin was suspended vehicle and
administered at the dose of 150 mg/kg of body weight.
III Suspension of test substances: Selected formulation were suspended in vehicle
and administered at their decided dose levels.
In vivo screening of different combinations for oral glucose
tolerance test in normal rats.
The oral glucose tolerance test for different selected combinations were
performed in overnight (18 hr) fasted normal rats.
Glucose (3 g/kg) was fed orally to each group 30 min after the administration
of combinations and Metformin. Then each group was subjected to blood
collection for 60,90,120 min.
23. 24
Collection of blood and estimation of serum glucose:
Blood was withdrawn using haematocrit capillary from the retro orbital plexus
under ether inhalation anesthesia at -30, 0, 30, 60, and 120 min of glucose
administration and glucose levels were estimated using glucose oxidase-
peroxidase standard kit from Span Diagnostics Ltd. India.
Estimation of glucose
Calculation
Plasma Glucose mg/dl =
Absorbance of Test
Absorbance of
Standard
* 100
26. 27
From the above study Combination 2 was showing good blood glucose
lowering activity( Anti-Hyperglygemic) compared to other combinations
during the OGTT study.
Combination 2 was showing decrease in glucose level at the time interval of 60
min .
Hence for further development in formulation and streptozotocin induce Anti-
Diabetic study this combination along with optimized formulation were
selected.
27. Future studies
28
Optimization of excipients used in the formulation.
Method of preparation of herbal tablet
Evaluation of herbal tablet as per WHO guideline.
Phytochemical analysis of the herbal tablet by
HPTLC.
STZ-NAD induced Anti-Diabetic evaluation of the
selected combination and optimized formulation.
