EVALUATION OF NOOTROPIC ACTIVITY OF ETHANOLIC EXTRACT HIBISCUS ROSA SINENSIS.
M.PHARM DISSERTATION PROTOCOL SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,BANGALORE,
KARNATAKA.
BY
MOHD. ISMAIL ZUBAIR
B.Pharm
UNDER THE GUIDANCE OF
Prof. SHASTRI V.V. _
M.Pharm Ph.D
DEPARTMENT OF PHARMACOLOGY
KARNATAKA COLLEGE OF PHARMACY
BIDAR-585403
2011-2012
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.0
/NAME OF THE CANDIDATE AND ADDRESS(IN BLOCK LETTERS)
/MOHD. ISMAIL ZUBAIR
S/O MOHD HASHAM ALI
H.NO : 5/3/147(N), GOLE KHANA,
OPPOSITE OLD DALL MILL.
BIDAR-585401
2.0
/NAME OF THE INSTITUTION
/KARNATAKA COLLEGE OF PHARMACY BIDAR-585403
3.0
/COURSE OF STUDY AND SUBJECT
/MASTER OF PHARMACY IN
PHARMACOLOGY
4.0
/DATE OF THE ADMISSION
/14/07/2011
5.0
/TITLE OF THE TOPIC:
“Evaluation of Nootropic Activity of Ethanolic Extract Hibiscus Rosa Sinensis”
6.07.0 / BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDY
Memory is ability of an individual to record event, information and retains them over short or longperiods of time and recalls the same whenever needed. Age, stress and emotion are conditions that may lead to memory loss, amnesia, anxiety, high blood pressure, dementia, to more ominous threat like schizophrenia and Alzheimer’s diseases. Alzheimer’s disease is progressive neurodegenerative disease that primarily affects the elderly population, and is estimated to account for 50-60% of dementia cases in persons over 65 years of age1, 2. “Nootropics” are agents that enhance the cognitive skills, and “amnstics” are agent that disrupts the learning and memory processes. Learning and memory can be conceived as both a psychological process, as well as a change in synaptic neural connectivity3,. Cognitive deficits have long been recognized as severe and consistent neurological disorders associated with numerous psychiatric and neuro-degenerative stats. Indian systems of medicine emphasize use of herbs, nutraceuticals or life style changes for controlling age related neurodegenerative disorders.
6.2 REVIEW OF LITERATURE:
6.2.1Plant review12
Botanical name: Hibiscus rosa sinensis
Family name: Malvaceae
Synonym: Sanskrit japa, Hindi jasum, Bengali jaba, English Chinese hibiscus, Bunga Raya in Malayalam,Sembaruthi in Tamil and Dasananu in Telugu.
Hibiscus Rosa Sinensis(Family: Malvaceae) is widely cultivated in many parts of India. Phytochemical studies revealed the presence of several chemicals including flavonoids, hibiscetin, cyanidine, cyanin glycoside, taraxerylacetate,β-sitosterol,campesterol,stigmasterol,ergosterol, citric, tartaric and oxalic acids cyclopropenoids and anthocyanin pigments .Theplanthas also been evaluated pharmacologically for the presence of anticonvulsant , antitumor , hypoglycaemic and estrogenic activities.In view of the traditional uses ofHibiscusrosasinensisinconstipationand diarrhoea, the crude extract of aerial parts and its subsequent fractions were tested on isolated intestinal preparations to provide pharmacological basis for their folk use in the gut motility disorders, constipation and diarrhea, anti complementary, anti-diarrhetic, anti-phologistic, anti-spermatogenic , androgenic ,anti-tumour and anticonvulsant4 hair growth5 and anti ulcer6
6.2OBJECTIVE OF THE STUDY
- To collect the flowers of the plants and authenticated
- To extract the dried powered of the flower with absolute ethanol
- To carry out preliminary phytochemical analysis of ethanolic extract.
- To carryout toxicity studies of ethanolic extract of flower of Hibiscus rosa sinensis
- To asses anti oxidant property (In-Vitro)
- To evaluate the nootropic activity of Hibiscus rosa sinensis extract by using experimentally induce disruption of memory (amentia) by differing models.
2)Elevated plus maze (Transfer latency) and its disruption by Scopolamine and MES were used as two induction method.
MATERIAL AND METHODS
7.1 Source Of Data :
The standard information is collected from various journals, standard Text books available in library of Karnataka college of pharmacy, Indian Institute of Science, RGUHS digital library and from various standard websites.
Web sites:
7.2 METHOD OF COLLECTION OF DATA:
a) Collection and extraction of plant materials
The Hibiscus rosa sinensis flower is collected in and around the BIDAR district. and authenticated by the Botanist. About 500grms of dried flowers are extracted successively with solvent by using soxhlet apparatus under reduced pressure and controlled temperature. The prepared extract will be concentrated under reduced pressure and stored in air tight container.
b) Toxicity studies
Acute oral toxicity study is to be carried out for the determination of LD50 by using OECD guide line- 423
c)In Vitro anti-oxidant activity11
1)Reducing power
2) Superoxide anion scavenging activity.
3) Hydroxyl radical scavenging activity.
d) NOOTROPIC ACTIVITY
1. Evaluate of nootropic activity of Hibiscus rosa sinensis.
ANIMALS REQUIREDa) Species : Wistar albino rats
b) Weight : 150-220 gm
c) Gender : Either sex
d) Number : 96 rats
Groups of Animals :
Group 01: (n=6) Normal Control Treated with vehicle
Group 02: (n=6) Positive Control – Treated with Extract of Hibiscus Rosa Sinensis
Group 03: (n=6) Positive Control - Treated with Mentat(Standard Drug).
Group04: (n=6) Negative Control – MES Induced.
Group 05: (n=6) Nagative Control – Scopolamine Induced.
Group 06: (n=6) Tratment Group- MES + Treated with extract of Hibiscus Rosa Sinensis
Group 07: (n=6) Tratment Group- MES+ Treated with Mentat (Standard drug).
Group 08: (n=6)Treatment Group – Scopalamine + Treated with Extract of HibiscusRosa Sinensis
Group 09: (n=6) Treatment Group – Scopalamine + Treated with Mentat (Standard Drug).
Experimental Schedule :
All the animals were dosed once in a day with respective drug for ten days. All drugs were given through per oral route (p.o) Group-1 animals received only the vehicle(0.5% tween 60), Group-2 animal received Extract Hibiscus rosa sinensis. Group-3 received only mentat(standard Drug)8 Group 4 received only electric shock (150mA for 0.2s), Group-5 received only scopolamine(0.3 mg/kg b.w.)7, Group-6 and Group 7 recieved both electric shock and respective drug treatments,Group-8 received scopolamine and Extract HibiscusRosa Sinensis was administered orally Group-9 received scopolamine and Mentat.
The animals were trained on the 0(zero) day and the acquisition of memory was tested on the day 1, later the animals were subjected to induction followed by drug treatment, that was continued for upto day 10. Then the animals were subjected for the retention test on the day 10.
Transfer latency :The rats were placed individually at the end of one arm facing away from the center of the maze and the time the rat took to move from open arm to either of the enclosed arms (Transfer latency, TL) was recorded. On the first day, the rats were allowed to explore the plus maze for 20sec after the measurement of TL. Rats were returned to their home cages after the first trial. Twenty four hours later, the rats were placed on the elevated plus-maze individually as before and TL was recorded again. TL measured on 1st and 2nd day served as parameters for acquisition and retrieval respectively. All the drugs were administered 30min to the first trial, either alone or in combination and each treatment group consisted of 6 animals9.Passive Avoidance Task (step-down latency): The procedure described by Vasudevan Raghavendra, Shrinivas K. Kulkarni.10 was used. In brief, the apparatus consisted of an electric grid (24 x 30 cm) with a shock-free zone (SFZ; 2 x 3 x 1 cm) in the centre and the entire grid having a perflex enclosure10.
e) Statistical analysis
The data obtained from the above findings will be subjected to statical analysis using one-way ANOVA followed by student-t-test
f) Work of plan details :
Total duration for the completion of proposed research work may be 10 months.
- Authentication and collection of plant material – 1 month
- Duration of experimentation on animals including preparation of crude extract- 5 month
- Literature review- 2 month
- Dissertation writing – 2 month
APPLICABLE
ANIMALS:
Wistar albino rats of either sex (weight: 150-250gm) will be involved in this study. The animals are kept in cage and maintained under standard housing condition (room temperature 24-270C) with 12hr dark and light cycle. The food in the form of dry pellets and water will be available. The experiment will be performed as per CPCSEA guidelines.
7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
Yes, Ethical clearance certificate is attached with hard copy.
LIST OF REFFERENCES:
1.Sunil n, kshirsagar. Nootropic activity of dried seed kernels of caesalpinia crista linn against scopolamine induced amnesia in mice. International journal of pharm tech research 2011; 3(0974-4304):104-409.
2.Hanumanthachar joshi, milind parle. Zingiberofficinale:evaluation of its nootropic effect in mice. Afr. J. Traditional, complementary and alternative medicines 2006; 3(0189-6016):64-74.
3.Baburao chandu, mukkanti khagga, silva r challa, varun dasari. Evaluation of nootropic potential and in vitro antioxidant activity aqueous extract of roots of asparagus racemosus in rats international journal of pharmaceutical research and developments 2011; 3(0974-9446):184-191.
4.Anwar h. Gilani, samra bashir, khalid h. Janbaz, abdul jabbar shah. Presence of cholinergic and cacium channel blocking activities explains the traditional use of hibiscus rosasinensis in constipation and diarrhoea. Journal of ethnopharmacology 2005; (102):289-294.
5.Adhirajan n, ravi kumar t, shanmugasundaram n, mary babu. In vivo and in vitro evaluation of hair growth potential of hibiscus rosa-sinensis linn. Journal of ethnopharmacology 2003; 88:235-239.
6.Rajesh j mandade, sreenivas sa, sakarkar et al dm. Pharmacological effects of aqueous-ethanolic extract of hibiscus rosasinensis on volume and acidity of stimulated gastric secretion. Asian pacific journal of tropical medicine 2011; :883-888.
7.Meera sumanth, sowmya h, nagaraj sv k. Efficacy of donepezil and galantamine in retrograde amnesia. Asian journal of pharmaceutical and clinical research vol. 2010; 3(0974-2441):25.
8.Bhattacharya sk, nootropic effect of br-16a (mentat), a psychotropic herbal formulation, on cognitive deficits-induced by prenatal undernutrition, postnatal environmental impoverishment and hypoxia in rats. [indian journal of experimental biology 1994; 32(1):1,31.
9.Kulkarni sk, anita verma. Evidence for nootropic effect of br-16a (mentat), a herbal psychotropic preparation, in mice. Indian journal of physiology and pharmacology 1992; 36(1):1,29.
10.Vasudevan raghavendra, shrinivas k. Kulkarni. Possible antioxidant mechanism in melatonin reversal of aging and chronic ethanol-induced amnesia in plus-maze and passive avoidance memory tasks. Free radical biology & medicine 2000; 30(6):595-602.
11.Chanda S, Dava R. In- vitro model for anto- oxidant activity evaluation and some medicnal plant possessing anti-oxidant properties. African journal of microbialogical Research. 2009: 3.13 : 981-96.
12.Sukirti Upadhyay, Prashant Upadhyay. Hibiscus rosa sinensis : pharmacological review. internationl j of research in pharmaceutical and biomedial sciences 2011; 2(4)(2229-3701):1449-1450.
9.0
/SIGNATURE OF THE CANDIDATE
/MOHD. ISMAIL ZUBAIR
10.
/REMARKS OF THE GUIDE
11.
/NAME AND DESIGNATION
11.1 GUIDE
/Mr.SHASTRI V. V
Professor
DEPT. OF PHARMACOLOGY,
KARNATAKA COLLEGE OF PHARMACY
BIDAR-585403
11.2 SIGNATURE
11.3 CO-GUIDE (IF ANY)
/------
11.4 SIGNATURE
/------
11.5 HEAD OF THE DEPARTMENT
/Mr. RAGHAVENDRA PATIL.
H.O.D;
DEPT. OF PHARMACOLOGY,
KARNATAKA COLLEGE OF PHARMACY
BIDAR-585403
11.6 SIGNATURE
12.
/12.1 REMARKS OF THE PRINCIPAL
12.2 SIGNATURE
/Prof. Dr.SIRSE KRANTIKUMAR
PRINCIPAL,
KARNATAKA COLLEGE OF PHARMACY
BIDAR-585403