Pankaj B. Magadum

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Comparative study of sildenafil, Chlorophytum borivilianum leaf and root aqueous extracts on aphrodisiac activity and its influence on cardiac profile.”


Pankaj B. Magadum. B Pharm

Dissertation Protocol Submitted to the

Rajiv Gandhi University of Health Sciences,

Bangalore – 560040, Karnataka.

In partial fulfillment

Of the requirement for the degree of



Under the Guidance of

Dr. Benson Mathai K, M.Pharm,Ph.D.


Dept. of Advanced Pharmacology and Toxicology

St. John’s Pharmacy College.







Name of the candidate and address







Name of the institution

St. John’s Pharmacy College

No 6,9th cross, 2nd Main, Vijaynagar 2nd Stage (Hampinagar)

Bangalore- 560104

Tel: 91-80-23300958/23300668




Course of study and subject



Date of the admission

22th JUNE 2010


Title of the topic:

“Comparative study of sildenafil, Chlorophytum borivilianum leaf and root aqueous extracts on aphrodisiac activity and its influence on cardiac profile.”

6. Brief resume of the intended work:

6.1 Need for the study:

Sexual activity is an important component of a human well being and quality of life, Sexual dysfunction (impotence) and other related problems is eluding scientific community and medical practitioners since time immemorial.[1] Erectile dysfunction may affect 30-50% of men aged 40-70 years. [2]

Presently, pharmaceutical compound sildenafil citrate (Viagra®) is looked upon as a reliable drug to treat sexual dysfunction. But due to venodilator effects on the peripheral vasculature it has got undesirable side effects specially on cardiac activity in patients having history of cardiac problems.[3] The risk association between erectile dysfunction (ED) and new onset of coronary artery disease (CAD) has been documented in a recent study, which strongly suggested that ED is a surrogate marker for future CAD.[4] There has been a constant exploration for newer herbal and chemical agents to overcome these age-old problems of sexual dysfunction with less side effects.[1]
The present study is designed to validate on scientific basis for traditional use of the plant Chlorophytum borivilianum (CB) for treating sexual dysfunction and also will check for its action on cardiac profile as compared to sildenafil citrate.

    1. Review of the Literature:

The Chlorophytum has 256 varieties in the world, 17 of them are found in India, amongst which C. borivilianum (CB) belonging to the family Liliaceae is indogenous medicinal plant which is cultivated in eastern parts of India and has good market demand. The common name of this herb is SAFED MUSLI. [5] It possess unique healing and health giving properties which makes it useful as a tonic and an important ingredient in around 20 ayurvedic and unani medicines of which CHYAWANPRASH is the best example. [6]

Extracts of tubers has been studied for its antidiabetic, antistress, immunomodulatory,anti inflammatory, antioxidant, anti microbial and anthelmintic activity.[7] Fasiculated roots of C.borivilianum are used in preparation of medicines.

Dry safeed musli contains carbohydrates, proteins, fiber, saponins (contains sapogenins, stigmosterols herbal steroids, hecogenin) which are responsible for its aphrodisiac activity[8] and alkaloids which are the primary source of its significant medicinal properties. Because of its saponin content CB is one of the plant which have known to possesses aphrodisiac potential traditionally and has numerous reference in Ayurvedic Materia Media. Stigmasterol, the chief constituent of Chlorophytum borivilianum is structurally similar to that of corticosteroids and testosterone and hence known to enhance blood levels of these sex hormones.[5]

There are many reported studies on aphrodisiac activity of various herbal extracts in the literature. Thakur et al reported that the aqueous extract of Dactylorhiza hatagirea (D.Don) possesses the considerable effect on the sexual behavior an performance on male rats.[9] Emilia et al showed the aphrodisiac property of the ginseng in rats.[10] Alexis et al showed that Caesalpinia benthamiana roots have the aphrodisiac property in male rats.[11] Zonali et al proved the significant effect of Eurycoma longifolia on the copulatory behaviour and sexual impotence in rats.[12]

The aphrodisiac activity of CB root has been proved scientifically.[13] But no study has been carried out on leaves of CB for the said activity. Therefore this study has been carried out to validate aphrodisiac activity of leaves of CB on scientific basis

    1. Objectives of the study :

The objectives of the present study are as follows :

  1. To evaluate the Aphrodisiac activity of aqueous extract of CB leaf.

  2. To compare the aphrodisiac potential of aqueous extract of CB leaf and root in comparison with sildenafil citrate.

  3. To study the cardiac effect of aqueous extracts of CB leaf and root in comparison with sildenafil citrate.

7. Materials & Methods

7.1 Source of data

Whole work is planned to generate data from laboratory studies i.e.; experiments are performed as described in reference, experimental studies in journals and in textbooks available with college, Indian Institute of Sciences (IISc) library, various other institutions of Bangalore and Rajiv Gandhi University of Health Sciences (RGUHS) digital library (Helinet).

Websites:,, etc.

were used to obtain related information regarding this research protocol.

7.2 Chemicals- All chemicals of standard quality and grade will be procured.

7.3 - Method of collection of data:

The data collected will be based on laboratory animal experimentation. The experiments will be conducted using laboratory animals and the data will be collected by analyzing the appropriate parameters. All experiments are planned in accordance with Committee for the purpose of control and supervision of experiments on animals (CPCSEA), Chennai, India. The present study has got the Institutional Animal Ethical Committee (IAEC) approval (certificate attached).

  1. Experimental animals:

  • Species: Albino Wistar Rats

  • Age: adult

  • Weight: 225-250

  • Gender: Male

  • Number: 36.

  • Duration of housing: 4-5 months

30 female rats from breeding stock will be used for mating purpose only and will be returned.

The animals will be collected from Central animal facility, St. Johns Pharmacy College, Bangalore, Karnataka, India. The animals will be housed in sterile polypropylene cages containing sterile paddy husk as bedding material with maximum of 3 animals in each cage. The experimental procedures will be performed between 10.00 and 16.00hrs. The mice will be fed on autoclaved standard mice food pellets and water ad libitum. Animals which do not comply with above criteria and which are found to be diseased will be excluded from the study.
(b)Acute oral toxicity study[14]:

Acute toxicity study for the aqueous leaf extracts of CB will be done according to the OECD guidelines No:423 and low and high dose will be selected for treatment.

(c)Preparation of drug:

The dry aqueous extract obtained will be soluble in distilled water 2–3 h prior to experimental

use to obtain the desired concentrations ( low dose and high dose ) in 1 mL.

Mode of administration of Chlorophytum borivilianum extracts: The test drug and the standard
drug will be administered to the respective groups orally by gavage once daily for 28 days. The standard
drug, sildenafil citrate (4mg/kg body weight) will be prepared in 0.1% sodium carboxy methyl cellulose
suspension for oral dose.

(d)Group classification:
Group 1 : Control group treated with distil water

Group 2 : CB leaf extract at the low dose.

Group 3 : CB leaf extract at the high dose.

Group 4 : CB root extract at the low dose

Group 5 : CB root extract at the high dose

Group 6 : Viagra® group: 4 mg/kg/day sildenafil citrate

(Dose of the Viagra is based on previous published article[1]
Treatment period for all these groups will be 28 days [Duration of treatment was fixed based on previous published article].[1]


  • sexual behavior analysis[1,15]

  • effect on sexual organ weight[1,18]

  • penile erection index[1,16]

  • orientation behavior analysis[1,16]

  • Sperm analysis[1,17]

The above mentioned parameters will be evaluated on days 1, 7, 14, 21, and 28 of treatment by pairing with a pro-estrous ovariectomized female rats.

This study is designed to find out the effect of leaf and root aqueous extracts of CB on the cardiac activity in comparison with the standard drug sildenafil citrate.

Sildenafil citrate has been reported to show cardiac dysfunction on first day itself in clinical studies.[19] In the present study cardiac activity will be evaluated on days 1,7, 14, 21 and 28 of treatment.


  • Electrocardiogram(ECG)

  • Serum glutamate oxaloacetate transaminase ( SGOT),

  • Lactose dehydrogenase (LDH)

The ECG will be recorded using polywriter on the days of 1st ,7ths 14th, 21st and 28th day of the drug treatment.

The following parameters will be estimated at the end of the treatment period (28 days) after sacrificing the rats by isolating the heart.

  • SGOT

  • LDH

The above enzyme estimation will be carried out by using biochemical kits in a semi-autoanalyser.


  • Lipid peroxidation (LPO)

Thio barbituric acid reactive substancess (TBARS) in the heart homogenate will be estimated using the standard protocol.[20]

  • Super oxide dismutase (SOD)

The assay of SOD will be carried out based on standard procedure.[21]

Statistical analysis:

Values will be expressed as mean ± SEM from 6 animals. Statistical difference in mean will be analyzed using one way ANOVA (analysis of variance) with suitable post test.

Number Of Animals Required:

    • No of the animal in each group = 6

    • No of groups = 6

    • Total no. of male animals required = 6 × 6 = 36

    • Total no of animals = 36

Accepted October 26th, 1972)EHAVIOR OF MALE CA

7.4 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly?

Yes, the study requires investigation on male wistar rats.

7.5 Has ethical clearance been obtained from your institution in case of 7.3?

Institutional Animal Ethical Committee (IAEC) clearance has been obtained.

8. References:

  1. Vikas S, Mayan T, Nagendra S.C, Vinod K.D.. Evaluation of the anabolic, aphrodisiac and reproductive activity of Anacyclus pyrethrum dc in male rats. sci pharm 2009; 77: 97–10.

  2. Prathap T, Ganesh G. Erectile dysfunction Mens’ Health. Clinicalevidence 2006 :1-30. Avaible at URL:

  3. Melvin DC, Adolph MH, Johnson R, Ralph GB, Peter G, Sanjay K, et al. Use of sildenafil (viagra) in patients with cardiovascular disease. Circulation 1999;99:168-77.

  4. Ajay N. Erectile dysfunction and cardiovascular disease:efficacy and safety of phosphodiesterase type 5 inhibitors in men with both conditions. Mayo Cli. Proc 2009;84:139-48.

  5. Guno SC, Vidhu A. Phytochemical and antimicrobial studies of Chlorophytum borivilianum. IJPSDR 2009 ;1:110-12.

  6. Haque R, Saha S, Roy A, Bera T. Micropropagation of an medicinal plant Chlorophytum

Borivilianum through shoot crown bud and characterization. Int J Ph Sci 2009;1:250-60.

  1. Deore Sl, Khadabadi SS. Invitro anthelmintic study of chlorophytum borivilianum sant and fernandez tubers. Indian J Nat Prod Research 2010;1:53-56.

  2. Nutan K. Saponins of Chlorophytum species. Phytochemistry Reviews 2005;4:191-96.

  3. Thakur M, Dixit VK. Aphrodisiac Activity of Dactylorhiza hatagirea (D.Don) Soo

in male albino rats. eCAM 2007;4(S1)29–31.

  1. Emilia N, Marianna A, Angelo A, Izzo. The aphrodisiac and adaptogenic properties of ginseng. Fitoterapia 2000:1-5.

  1. Alexis Z, Franc N, Sevser S, Thierry H, Bart S, R´egis B. Vasoactivity, antioxidant and aphrodisiac properties of Caesalpinia benthamiana roots. J Ethanopharmacol 2008;116: 112–19.

  2. Zanoli, P, Zavatti M, Montanari C, Baraldi M. Influence of Eurycoma longifolia on the copulatory activity of sexually sluggish and impotent male rats. J Ethanopharmacol 2009;126:308–13.

  3. Rakesh K, Riddhi S, Sadhana S. Effects of Chlorophytum borivilianum on sexual behavior and sperm count in male rats. Phytother Res 2008;22:796-01.

  4. The organization of economic co-operation and development (OECD) the guideline for testing of chemicals: 423 acute oral toxicity, OECD, Paris 2001;1-14

  5. Anders A. Male rat sexual behavior. Brain Research 1997;1:203-09.

  6. Ageel AM, Islam MW, Ginawi OT, Al-Yahyat MA. Evaluation of the aphrodisiac activity of Litseachinensis (Lauraceae) and Orchis malculata (Orchidaceae) extracts in rats. Phytother Res 1994 ;8:103-05.

  7. Rakesh K, Riddhi S, Sadhana S. Effect Of Chlorophytum borivilianum On Sexual Behavior And Sperm Count In Male Rats. Phytother Res 2008;22:796-01.

  8. Suresh S, Prithiviraj, Prakash S. dose and time dependent effects of ethanolic extract of Mucuna pruriens linn.seed on sexual behavior of normal male rats. J ethanopharmacol 2009;110:497-501.

  9. Jeppe GR, Egon T, Ole F.E Ventricular tachycardia after administration of sildenafil citrate: a case report. JMCR 2007;1:65.

  10. Gelvan D, Saltman P. Different cellular targets for Cu- and Fe-catalysed oxidation observed using a Cu-compatible thiobarbiturate acid assay, Biochim Biophys Acta 1990;1035:353-60.

  11. Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-5.


Department of Anatomy, School of Veterinary Medicine, University of California, Davis, Calif. 95616


Signature of the candidate

( pankaj magadum)


Remarks of the Guide:


Name and Designation of:

11.1 Institutional Guide

Dr. Benson Mathai K, M.Pharm , Ph.D

Professor & Principal


Department Of Pharmacology And Toxicology,

St. Johns Pharmacy College


11.2 Signature

11.5 Head of the Department

11.6 Signature

Dr. Girish Gowda, M.Pharm , Ph.D

Asst. Professor &

Head Of

Department Of Pharmacology And Toxicology,

St. Johns Pharmacy College



12.1 Principal

Dr. Benson Mathai K

Professor & Principal

Department Of Pharmacology And Toxicology,

St. Johns Pharmacy College


12.3 Remark of the principal

12.4 Signature

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