DEVELOPMENT AND CHARACTERISATION OF HERBAL FORMULATION

OF ARTEMISIA ANNUA AND MORINDA CITRIFOLIA LINN

M. Pharm Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore – 560041

By

Mr.KARNATI NITHIN KUMAR,B.Pharm

Under the Guidance of

Mr. SATEESHA .S.B

Asst.Professor

Department of Industrial Pharmacy

AcharyaB.M.Reddy College of Pharmacy

Soldevanahalli,Chikkabanavara (Post),

Hesarghatta main road, Bangalore – 560 090

2010– 2012

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE.

ANNNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1 / Name and address of candidate / Mr. KARNATI NITHIN KUMAR
H.No- 5-110, Rajyalakshmi Nagar 2,
Gurramguda,Saroornagar(M),Hyderabad,
Ranga Reddy (Dist), Pin 501510.
ANDHRA PRADESH.
2 / Name of institution / ACHARYA & B.M. REDDY COLLEGE OF
PHARMACY.
Soldevanahalli, Hesarghatta Main Road,
Chikkabanavara Post,
Bangalore-560090.
3 / Course of study and subject / M. Pharm
(Industrial Pharmacy)
4 / Date of admission /
18-09-2010
5 / Title of the project / DEVELOPMENT AND CHARACTERISATION OF HERBAL FORMULATION OF
ARTEMISIA ANNUAAND MORINDA CITRIFOLIA LINN
6
6.1 / BRIEF RESUME OF INTENDED WORK
NEED FOR THE STUDY:
A Gastro intestinal inflammatory disease primarily affects the digestive tract (GI tract) and any area of the GI tract from the mouth to the anus. It most commonly affects the lower part of the small intestine, such as ileum and jejunum. The swelling extends deep into the lining of the affected organ, causing abdominal pain, diarrhoea or constipation, malnutrition and weight loss. It is difficult to diagnose and its etiology is not clear. Types of inflammatory GI disordersare Crohn's disease and ulcerative colitis as these are inflammatory bowel diseases(IBD). Crohn’s disease isan auto immune disease in which all layers of the intestine may be involved leaving the body unprotected from infection. Researchers believe that the immune system mistakes bacteria or foreign invading substancesthat is normally found in the intestines and launches an attack sending white blood cells into the lining of the intestines, where they produce chronic inflammation. These cells then generate harmful products that ultimately lead to ulcerations and bowel injury. When the small intestine is inflamed, its ability to fully digest and absorb nutrients from food is reduced. These nutrients and unabsorbed bile salts, can escape into the large intestine which can cause a loss of appetite, diarrhoea, dehydration and malnutrition. If the large intestine also becomes inflamed, the diarrhoea may become even more extreme. Ulcerative colitis causes inflammation and ulcers in the top layer of the lining of the large intestine1.
Optimal treatment of inflammatory bowel disease which are currently available are depends on what form it consists of, for example,mesalamineis more useful in ulcerative colitisthan inCrohn's disease.Generally, depending on the level of severity, IBD may requireimmune suppressionto control the symptom, such asprednisone,Tumour necrosis factor (TNF), Azathioprine methotrexate, or6-mercaptopurine. More commonly, treatment of IBD requires a form ofmesalamine. In use for several years in Crohn's disease patients and recently in patients with ulcerative colitis,biologicalshave been used such as TNF inhibitors. Usually the treatment is started by administering drugs with high anti-inflammatory effects, such as prednisone2.
As demerits of these existing medications for biologicals are lymphoma, infections, congestive heart failure, lupus like syndrome, injection site reactions, systemic side effects. Using the high anti-inflammatory drugs continuously causes diarrhoea, nausea, cramping. Using the steroids such as prednisone short term side effects as with all glucocorticoids are increased blood glucose especially in diabetic patients, increase in body weight, depression, blurred vision etc3.
The existing treatment can be improved by reducing the adverse effects of allopathic medications and/or using the herbal formulations. Synthetic drugsmay bring onwell-known and documented harmful, side-effects while natural herbs contain infinite intelligence; they usually have no harmful side-effects. Herbs are natural, they are more readilyabsorbed and assimilated by the body. Herbal drugs are considered less potent than prescribed medicines however; herbs will have several active ingredients that are chemically similar and contain manyknown and some still unknown synergistic ingredients.So, the objective of present research work is to design and development of herbal formulation for anti-inflammatory diseases using the stems ofArtemisia annua4and fruits ofMorinda citrifolia Linn5which are having a potent anti-inflammatory activity.
6.2
6.3 / REVIEW OF LITERATURE
  1. Oscar CTet al., studied the aetiology of inflammatory bowel disease (IBD) involves a combination of genetic predisposition and environmental factors (cigarette smoking, use of non-steroid anti-inflammatory drugs, psychological stress and the presence of the caecalappendix) have been postulated as a trigger of IBD. He has also been suggested thatthe gut microbiota plays a major role in the development and persistence of IBD, and numerous modifications of intestinal microbiota composition have been identified6.
  1. Balfour SRstudied the Crohn’s disease and ulcerative colitis is idiopathic, chronic, relapsing,inflammatory conditions that are immunologically mediated. His studies indicate that Crohn’s disease and ulcerative colitis are heterogeneous diseases characterized by various genetic abnormalities that lead to overly aggressive T-cell responses to a subset of commensal enteric bacteria and different genetic abnormalities can lead to similar disease phenotypes; these genetic changes can be broadly characterized as causing defects in mucosal barrier function, immune-regulation or bacterial clearance7.
  1. Udai PS et al., studied the rates of incidence and prevalence of Crohn’s disease and ulcerative colitis and his stem cell (SC) research has become a new direction for IBD therapy. His findings suggest that, in the future, SC-based therapy will be a promising alternative to conventional therapy for IBD8.
  1. Smita Net al., studied the physicochemical and phytochemical evaluation of Morinda citrifolia Linn fruit extractives. She also carried out phytochemical screening by qualitative chemical methods. Additionally, she developed fractionation methodologies and standardized for isolation of components rich in polysaccharides, anthraquinones and alkaloids. Her TLC studies confirmed the presence of scopoletin, an important marker in the identification of Morinda fruits9.
  1. Jagtap AG et al.,studied the polyherbal ayurvedic formulation against inflammatory bowel disease (IBD). He studied his formulations on two different experimental animal models of inflammatory bowel disease, which are acetic acid-induced colitis in mice and indomethacin-induced enterocolitis in rats. The formulation showed significant inhibitory activity against inflammatory bowel disease induced experimental animal models and results obtained established the efficacy of this polyherbal formulation against inflammatory bowel diseases10.
  1. Eng SOhas studied the botanicals and herbal preparations containing a single or two or more medicinal plants. He focussed on the analytical methodologies, which included the combination of sample preparation and chromatographic techniques for the chemical standardization of marker compounds or active ingredients in botanicals and herbal preparations. He has shown the applications of common chromatographic techniques, such as HPLC, CE, HRGC/MS, HPLC/MS and HPLC/MS/M for the analysis of compounds present in the plant extracts. He proposed the HPLC/MS procedures for the identification of marker or active compounds in plant extracts11.
  1. Yi-Zenget al., studied thedifferent chromatographic and electrophoretic techniques commonly used in the instrumental inspection of herbal medicines (HM). He recommended the chemical fingerprints obtained by chromatographic and electrophoretic techniques as a quality control tools for herbal medicine. He also reported the Hyphenated chromatographic techniques and chemometric evaluation as a powerful tool for quality control of herbal products12.
  1. Niharika Set al., studied the use of herbal drugs for the prevention and treatment of various health ailments has been in practice. They developed the effective marker systems for isolation and identification of the individual components and standardization, stability and quality control for herbal drugs13.
  1. Bina SS et al., studied the methanolic extract of the fruits of Morinda citrifolia, which showed antimicrobial, antioxidant, pesticidal, anti-HIV, anti-leshmaniasis and hypotensive activities. Their investigations focused on the isolation and characterization of a scopoletin constituent14.
OBJECTIVES OF THE STUDY
The objectives of the present study are following
Development of powder and syrup formulation using stems ofArtemisia annua (Asteraceae)and fruits of Morinda citrifolia Linn (Rubiaceae) extracts.
In vitro characterization of formulations.
Accelerated stability studies on the optimized formulations.
7.0
7.1
7.2 / MATERIALS AND METHODS
SOURCE OF DATA
1)Review of literature from:
  1. Journals
  • American journal of medicine.
  • Journal of Ethanopharmacology.
  • International journal of pharmacy and pharmaceutical sciences.
  • International journal of pharmacognosy and phytochemical research
  • Online journals.
b.Reference books
  • Quality control of herbal drugs by Dr. Pulok K. Mukherjee
  • Pharmacognosy by Trease and Evans 15th edition.
  • Plant drug analysis H.Wagner, S. Bladt, 2nd edition.
MATERIALS
DRUG: Stems ofArtemisiaannuaand fruits ofMorinda citrifoliaLinn.
SOLVENTS: Polar and non polar solvents etc.
POLYMERS: Biodegradable/non biodegradable polymers etc.
EXCIPIENTS: Approved pharmaceutical excipients as required etc.
METHODS
Collection and authentication of stems of Artemisia annua and fruits of Morinda citrifolia Linn.
Preparation of aqueous acetonitrile extract from stems of Artemisia annua15and aqueous extract of Morinda citrifolia Linn16.
Development of powder and syrup formulation using extracts.
In vitro characterization of formulation for
  • Micromeritic properties
  • Rheological properties
  • In vitro dissolution properties etc.
Accelerated stability studies on the optimized formulations.
.
METHOD OF COLLECTION OF DATA
Formulation of dry powder and syrup and its evaluation for the following parameters,Micromeritic properties, rheological properties andIn Vitro studies using pharmacokinetic models and any other study as required.
In vitro analysis of developed formulations and statistical analysis of the results.
Accelerated stability studies on the optimized formulations.
7.3

8.0
8.1 / DOES THE STUDY REQUIRE ANY INVESTIGATION TO BE CONDUCTED ON PATIENT OR OTHER HUMANS OR ANIMALS?
“ NO”
Has ethical clearance been obtained from your institution in case of 7.3?
“ not APPLIcable”
LIST OF REFERENCES
  1. Richard H,Georgina LH, Fiona LC, Emad M, Richard KR. Inflammatory bowel disease. JPaediatrchild h 2010; 20(10): 473-8.
  1. Treatment for crohns disease [homepage on internet]. 2011[updated 2011 May 12; cited 2006 July 30]. Available from.
  1. Byron C, Michael B. Gastrointestinal side effects of nonsteroidal anti-inflammatory drugs. Am J Med 1998;105(1): 33-40.
  1. Simone K, Talib NO, BilalO.Wormwood (Artemisia absinthium) suppressestumournecrosisfactoralpha and accelerateshealinginpatientswithCrohn’sdisease-Acontrolled clinical trial. J Phytomedicine 2010; 17(5):305-9.
  1. Douglas AK, Hee-byung C, Chung KS. The classical drug discovery approach to defining bioactive constituents of botanicals. J Fitoterapia 2011; 82(1):71-9.
  1. Oscar CT, Jose M, Angel G. Aetiology of inflammatory bowel disease (IBD): Role of intestinal microbiota and gut-associated lymphoid tissue immune response. J ESPEN 2005; 24(3): 339-52.
  1. Balfour SR.Mechanisms of Disease: pathogenesis of Crohn’s disease and ulcerative colitis. Jnatclinpractgastr 2006; 3(7):390 -407.
  1. UdaiSP, NarendraSP, Balwan S, Manoj KM, Mitzi N,Prakash SNet al., Stem cells as potential therapeutic targets for inflammatory bowel disease. Front Biosci (schol Ed) 2010; 1(2): 993-23.
  1. Smita N, sushmaM. Preliminary physicochemical and phytochemical evaluation of Morinda citrifoliaLinn fruit extractives. IJPPS 2010; 2(4): 150-4.
  1. Jagtap AG, Shirke SS, Phadke AS.Effect of polyherbal formulation on experimental models of inflammatory bowel diseases. J Ethnopharmacol 2004; 90(2-3):195-04.
  1. Eng SO.Extraction methods and chemical standardization of botanicalsand herbal preparations. J ChromatogrB 2004; 81(1-2): 23-33.
  1. Yi-Zeng L, Peishan X, Kelvin C. Quality control of herbal medicines. J ChromatogrB 2004; 812(1-2): 53-70.
  1. Niharika S, Padmavati M, SatyahariD. Herbal drugs: Sstandards and regulation. J Fitote 2010; 81(6): 462-71.
  1. Bina SS, Fouzia AS, Fayaz A,Sabira B. Isolation and Structural Elucidation of Chemical Constituents from the Fruits of Morinda citrifolia Linn. Arch pharm res 2007; 30(8): 919-23.
  1. Jain, Dharam CJ, Neerja P, Madan MG, Rajendra SB, Ram KVSet al., U.S patent: 6337095. Process for the isolation of the compound scopoletin useful as nitric oxide synthesis inhibitor, 2002.
  1. Sirima M, Wibool R,Sawpheeyah N, Narubodee P.Effects of Morinda citrifolia aqueous fruit extract and its biomarker scopoletin on reflux esophagitis and gastric ulcer in rats. J Ethnopharmacol, 2011; 134 (2): 243 - 50.

9 / Signature of the candidate:
10 / Remarks of the Guide:
11 / Name and Designation of:

11.1 Institutional Guide: / Mr.Sateesha S.B
Asst.Professor
11.2 Signature:
11.3 Co-Guide:
11.4 Signature:
11.5 Head of the Department: /
Mr. Anup Kumar Roy
Asst. Professor & HOD
Dept. of Industrial Pharmacy
11.6 Signature
12 / 12.1 Remarks of the Principal
12.2 Signature /
Dr. GoliDivakar
Principal
ACHARYA & B.M.REDDY COLLEGE OF PHARMACY,
SOLDEVANAHALLI,
HESARAGHATTA MAIN ROAD,
BANGALORE-90.