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Ocimum sanctum, anthocephalus cadamba, allium sativum, origanum vulgare


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DEVELOPMENT AND EVALUATION OF ANTIMICROBIAL OINTMENT

FORMULATION CONTAINING EXTRACTS OF OCIMUM SANCTUM,

ANTHOCEPHALUS CADAMBA, ALLIUM SATIVUM, ORIGANUM VULGARE.
M. Pharm Dissertation Protocol Submitted to



ZA
Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore - 560041.

By
Mr. T.RAJESH B.Pharm
Under the Guidance of

Prof. ANUP KUMAR ROY M.Pharm (Ph.D)




Department of Industrial Pharmacy,

Acharya & B.M. Reddy College of Pharmacy,

Acharya Dr. Sarvepalli Radhakrishnan Road,

Soldevanahalli, Chikkabanavara Post,

Bangalore - 560090.

2012- 2014.



RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BENGALURU, KARNATAKA.
ANNEXURE - II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION





1.


Name of the Candidate

and Address


MR. T.RAJESH,

S/O T.RAMANNA, H/NO 45/24/K63-1,

ASHOK NAGAR, KURNOOL,

ANDHRA PRADESH. 518001.




2.



Name of the Institution


ACHARYA & B.M. REDDY COLLEGE OF

PHARMACY, Acharya Dr. Sarvepalli Radhakrishnan Road,

Soldevanahalli, Chikkabanavara Post. Bangalore. 560 090.




3.


Course of Study and Subject



M. Pharm

(Industrial Pharmacy)




4.



Date of Admission



20 June-2012


5. TITLE OF THE PROJECT:-


DEVELOPMENT AND EVALUATION OF ANTIMICROBIAL OINTMENT FORMULATION CONTAINING EXTRACTS OF OCIMUM SANCTUM, ANTHOCEPHALUS CADAMBA, ALLIUM SATIVUM, ORIGANUM VULGARE.





6.0

7.0

8.0



BRIEF RESUME OF THE INTENDED WORK:

6.1 NEED FOR THE STUDY:

  • INTRODUCTION:



In the recent years, there has been a gradual revival of interest in the use of medicinal plants in developing countries because herbal medicines have been reported safe without any adverse side effect especially when compared with synthetic drugs. Thus a search for new drugs with better and cheaper substitutes from the plant origin are a natural choice1.

Herbal remedies for skin care with antibacterial and antifungal activities are prepared from a variety of plant parts such as leaves, stem, root, bark or fruit. These medicines are administered topically and may be applied in the form of cream, lotion, gel, soap, solvent extract or ointment, and have been establish to possess antimicrobial properties. Gels, creams and soap formulations containing a variety of plant extracts have been used to treat various skin disorders caused by microbial infections2.

Most of the skin infections are caused by fungi, Staphylococcus aureus and Streptococcus species. Treatment of bacterial infections is achieved by use of antibiotics, while fungal infections require antifungal pharmaceutical preparations such as clotrimazole solution2. Benefits of Herbal Cosmetics;

A number of people with sensitive skin don’t want to use chemical sunscreens due to concern about skin exposure to unknown chemicals. Topical cosmetic formulations are the most preferred treatments asked by patients and are also often most prescribed by family physicians and dermatologists. Patients feel more comfortable using topical therapies because they have milder side effects, are easier to use, are generally less expensive and are more readily available3.

The study of the present review focuses on the potential of herbal extracts for cosmetic purposes and the present disorder under study can be given enhanced therapy by reducing the adverse effects of allopathic medication by the use of natural herbs. Synthetic drugs may bring on well- known and documented harmful, side-effects whereas the use of natural herbs and their derivatives many have some synergistic effect which is not known. As herbs are natural products they would get ready absorbed and assimilated by the body, these contain several active topical ointment formulation for anti-microbial activity by the use of Ocimum sanctum, Anthocephalus cadamba, Allium sativum, Origanum vulgare3.




  • Ocimum sanctum (Tulsi)

Ocimum sanctum is one of roughly 60 species of the genus Ocimum, the basil genus which consist of aromatic herbs and shrubs indigenous to the topical regions of Asia and the America. It is also known as Holy Basil. They are 3 varieties of culmination are there 1) Rama Tulsi (Ocimum sanctum), Krishna Tulsi (Ocimum sanctum) and vana tulsi (Ocimum gratissimum). Tulsi applied topically as a paste with black pepper is used to treat ring worm and eczema.

Extensive studies, experimental and clinical prove that Tulsi possess anti-stress/adatogenic, antioxidant, immunomodulator and anti-radiation properties which also may help it to play major role in prevention and treatment of cancer. As Tulsi has health benefitting effects by reducing stress and improving both cellular and humoral immunity, its role in prevention and treatment of cancer cases may be a new approach in therapy of cancer and in prevention of ill effects of radiation4.



  • Anthocephalus cadamba (Kadamba)







Kadamba consists of dried stem bark of Anthocephalus cadamba (Rubiaceae) is ethno medicinally widely used in the form of paste by tribe in western Ghats for treating skin diseases. In this context, antimicrobial potential of A. cadamba against a wide range of microorganisms was studied. To validate the ethnotherapeutic claims of the plant in skin diseases, wound healing activity was studied, besides antioxidant activity to understand the mechanism of wound healing. The alcoholic and aqueous extract of this plant showed significant antibacterial and antifungal activity against almost all the organisms5.


  • Ilium sativum (Garlic)

Garlic has been used as both a food and a medicine and it is used externally to prevent wound infections. Louis Pasteur was one of the first scientist to confirm that garlic had antimicrobial properties. garlic has been used as a warming and blood cleansing herb to prevent and treat colds and flu, coughs, and parasites. Garlic appears to exert both direct and indirect effects against various pathogens as it enhances macrophage and T-lymphocyte function and has direct antibacterial, antifungal, antiviral and antiparasitic activity. Allicin is believed to be chiefly responsible for garlic’s antimicrobial activity6.


  • Origanum vulgare (Oregano)

Oregano is a perennial plant. It can grow to 80 centimeters in height. Leaves are dark green, oval and opposite. Flowers are white, pink or purple, formed in erect spikes. Oregano has long been used as a medicinal herb in treatment of various conditions: fevers, diarrhea, indigestion, jaundice and vomiting. Recent studies have shown that Oregano displays anti-oxidant, anti-fungal and antibiotic properties. Due to its anti-oxidant functions, Oregano could become helpful agent in treatment of cancer, heart disease and high blood pressure. It is also useful as a digestive aid, since it promotes salivation. Used externally, Oregano is successful in treatments of rheumatism, muscle and joint pain, sores and swellings7.

6.2 Objective of the Study

Following are the objectives of the present study ;


  • Collection of the different plants and authentication.




  • Extraction of selected plants by using soxlet apparatus.




  • Preliminary physiochemical investigation of different extract of the plants.




  • Formulation of the antimicrobial ointment by using extracts.




  • To carry out physicochemical parameters.

  • Measurement of pH

  • Acid value

  • Saponification value

  • Viscosity

  • Spreadability

  • Consistency




  • Antimicrobial evaluation




  • Bacterial strains and inoculum preparation.

  • Antimicrobial assay


MATERIALS AND METHODS:

7.1 Source of Data

  1. Journals such as,

    1. International Journal of Research in Cosmetic Science.

    2. Indian Journal of Pharmaceutical Sciences.

    3. Indian Journal of Pharmaceutical Education and Research.

    4. Drug Development & Industrial Pharmacy.

    5. African Journal of Pharmacy and Pharmacology.

    6. International Journal of Research in Pharmaceutical & Biomedical Science.

    7. Journal of Medicinal Plants Research.

    8. Asian General of Biomedical and pharmaceutical Science.

  2. Review articles.

  3. World Wide Web.J-gate@Helinet.

  4. Science Direct, Pub med and biomed central.

  5. Acharya & B.M. Reddy College of Pharmacy Library.


7.2 METHOD OF COLLECTION OF DATA:

MATERIALS

  1. Plant: The following plants can be used for preparing antimicrobial ointment.



Common name

Botanical name

Family

Part to be

Used

Tulsi

Ocimum sanctum

Lamiaceae


Leaves

Kadamba

Anthocephalus

Cadamba

Rubiaceae

Leaves

Garlic 

Allium sativum

Liliaceae

Pearls

Oregano

Origanum vulgare

Lamiaceae


Leaves









  1. Solvent: suitable solvents will be used.

  2. Excipients: suitable excipients will be used.


METHODS:

I. Collection of mature leaf from the local sources



  1. Tulsi leaf - collect 1kg of leaves, shade dry, weigh & then coarsely crush.

  2. Kadamba leaf - collect 1kg of leaves, shade dry, weigh & then coarsely crush.

  3. Garlic pearls - collect 100g & crush just before extraction.

  4. Oregano leaf - collect 1kg of leaves, shade dry, weigh & then coarsely crush

II. Extraction of the collected parts of the plant using different solvents

  1. Extraction of 100g of each dry herb with hexane, ethyl acetate, (80% methanol + 20% water) successively with 200mL solvent each time by using soxhlet apparatus.

  2. It is heated to boil & leave it at room Temperature overnight & then filter out (only one extraction) & this process is repeated for every extract.

  3. All solvent are to be stripped from each sample separately by using rotovapor under reduced pressure (Vacuum). It is to be kept at Keep water bath at max 600 C while evaporating & then take the extract in a 250mL Round Bottom flask & evaporate under reduced pressure to about 30mL & transfer to previously weighed 50mL Round Bottom flask. All solvents are to be stripped of and weight the extracts are to be taken. TLC studies will be carried out and documented.

III. Antimicrobial screening of collected plant extracts

  1. Two pathogens P1 (Gram +ve) and P2 (Gram -ve) will be taken.

  2. In-vitro screening with each of the 12 extracts individually and determine their MIC (minimum inhibitory concentration) and to be carried out.

V. Screening of the antimicrobial property of the prepared formulation

  1. Screening will be done for each formulation against the two pathogens and compare their MIC.

  2. Comparison with any one or two modern antibiotic suitable for (Gram +ve) and

(Gram –ve) bacteria.

VI. To carry out physicochemical parameters.



  • Measurement of pH

  • Acid value

  • Saponification value

  • Viscosity

  • Spreadability

  • Consistency


7.3 Does the study require any investigation or interventions to be

Conducted on patients or other humans or animals?
“NO”

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

“NOT APPLICABLE”



REFERENCES:

  1. Edeoga HO, Okwu DE and Mbaebie BO. Phytochemical constituents of some Nigerian Medicinal plants. Afr. J. Biotech. 2005; 4(7): 685-88.




  1. Kareru PG, Keriko JM, Kenji GM, Thiongo GT, Gachanja AH, Mukiira HN. Antimicrobial

activities of Skincare Preparations from Plant Extracts. Afr. J. Trad. Comp. Altern. Med. 2010; 7(3): 214-18.


  1. Tengamnuay P, prugsakij W, wongkajornsilp A. In Vitro Evaluation Of Anti -Ageing Activities Of Artocarpus Lakoocha Heartwood Extract. Afr. J. Trad. Comp. Alter. Med. 2008; 2(1): 826-28.




  1. Tulsi (HOLY BASIL) [Internet].[Cited on 10/12/2012] Available from http://www.organicindia.co.in/tulsi.php.



  1. Sanjay, Umachigi P, Kumar GS, Jayaveera KN, Kishore kumar DV, Ashok ck, kumar. Antimicrobial wounds healing and antioxidant activities of anthocephalus cadamba. Afr. J. Trad. Cam. 2007; 4(4): 481-87.



  1. Lau BH, Yamasaki T, Gridly DS. Garlic compounds modulate macrophages and T-lymphocyte functions. Mol. Biothepy.1991; 3(2): 103-07.




  1. Origanum vulgare [Internet]. [Cited on 29/11/2012] Available from: http://en.wikipedia.org/wiki/Oregano.




  1. Chhetri HP, Yogol NS, Sherchan J, Anupa KC, Mansoor S, Panna Thapa.

Formulation and evaluction of antimicrobial herbal ointment. Kathmandu. Univer. J. Scie. Engin and Tech. 2010; 6(1): 102-07.


  1. Chris A, Cecilia I, Chukwuemeka P, Azubuike. Evaluation of the Antibacterial activity of Herbal ointments formulated with Methanolic extract of Cassia alata. Asi. J. Biomed. Pharma. Scie. 2012; 2(13): 15-9.

  2. Rios JL, Recio MC, Villar A. Screening methods for natural products with antimicrobial activity. J. Ethnopharmacol. 1988; 23(3): 127-49.




  1. Rahman MM, Fazlic V, and Saad NW. Antioxidant properties of raw garlic (Allium sativum) extract. Inter. Food. Res. J. 2012; 19(2): 589-91.



  1. Kamkaen N, Soisuwan S, Weerasak S, Adelheid H. Development of peacock extract as anti- wrinkle formulation. J. Health. Res. 2010; 24(1): 29-34.



  1. Satish N, Damodar G, Atul D. Development and Evaluation of Antimicrobial Formulation containing extract of anthocephalus cadamba. Inter. J. Pharma. Res. Dev. 2011; 10(3): 8-12.




  1. Zakaria Ahmed. Ummeya and Allium Sativum. J. Pharm. Biol. Scie. 2012; 4 (1): 28-32.




  1. Kavitha T, Nelson R, Thenmozhi R, Priya E. Antimicrobial activity and phytochemical analysis of Anisomeles malabarica. J. Micro. Biotech. Res. 2012; 2(1): 1-5.




  1. Rees LP, Minney SF, Plummet NT, Slater JH and Skyrme DA. Quantitative assessment of the antimicrobial. World. J. Micro. Biotech. 1993; 9(1): 303-07.


  1. Harris JC, Cottrell S, Plummer S, Lloyd D, Antimicrobial properties of Allium sativum (garlic). Applied. Microbio. Biotech. 2009; 57(3): 282-86.



  1. Singh N, Verma P, Pandey BR, Bhalla M. Therapeutic Potential of Ocimum sanctum in Prevention and Treatment of Cancer and Exposure to Radiation. Inter. J. Pharma. Scie. Drug. Res. 2012; 4(2): 97-04.




  1. Nieblas MO, Burgueno RR, Felix EA, Leon AG, Trejo AM and Moreno LV. Chemical

Composition and Antimicrobial activity of Oregano (Lippia palmeri S. WATS) Essential oil.

Rev. Fitotec. Mex. 2011; 34(1): 11-17.





9.


SIGNATURE OF THE CANDIDATE







10.


REMARKS OF THE GUIDE





11.


NAME AND DESIGNATION OF









11.1 Guide


PROF. ANUP KUMAR ROY M.Pharm (Ph.D)


11.2 Signature






11.3 Head of the Department


Dr. ROOPA KARKI .

Professor & Head



Department of Industrial pharmacy




11.4 Signature






12.


12.1 Remarks of the

Principal



Forwarded to the University for approval





12.2 Signature




Principal

Dr. DIVAKAR GOLI M.Pharm, Ph.D

Professor and Principal

Acharya & B.M. Reddy College of Pharmacy,

Bangalore – 560090.







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