FORMULATION DEVELOPMENT AND VALIDATION OF AN ANTIMALARIAL HERBAL FORMULATION
M. Pharm Dissertation Protocol Submitted to
RajivGandhiUniversity of Health Sciences, Karnataka
Bangalore – 560041
By
Mr. SHAILENDRA SINGH KUSHWAHA,B. Pharm.
Under the Guidance of
Shri B. S. PATIL,M. Pharm. (Ph. D)
Post Graduate Department of Pharmacognosy,
SET’S College of Pharmacy,
S.R.Nagar, Dharwad,
Karnataka – 580 002.
RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
1 / NAME OF THE CANDIDATE AND ADDRESS / Mr. SHAILENDRA SINGH KUSHWAHAPost Graduate Department of
Pharmacognosy,
Set’s College Of Pharmacy,
S.R.Nagar,
Dharwad-580002
2 / NAME OF THE INSTITUTION / SET’S COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD-
Karnataka580002
3 / COURSE OF STUDY AND SUBJECT / MASTER OF PHARMACY IN PHARMACOGNOSY
4 / DATE OF ADMISSION TO COURSE / JUNE 2009
5 / TITLE OF THE TOPIC:
“FORMULATION DEVELOPMENT AND VALIDATION OF AN
ANTI MALARIAL HERBAL FORMULATION”.
6
7 / BRIEF RESUME OF THE INTENDED WORK
6.1 / Need for study
Malaria remains the world’s most devastating human parasitic infection, afflicting more than 500 million1 people and causing from 1.7 million to 2.5 million deaths each year. According to National Health Policy, 2002, the malaria cases in India in 2000 were 2.2 million. Malaria is commonly associated with poverty, but is also a cause of poverty2 and a major hindrance to economic development.
Malaria is a vector-borne infectious disease caused by a eukaryoticprotist of the genus Plasmodium3. In man, the infective protozoa are Plasmodium falciparum, P. vivax, P. malariae and P. ovale. P. falciparumcauses malignant tertian malaria and this is the most dangerous form of human malaria. P. vivax is the cause of being tertian malaria, in which the clinical attacks are milder than those of P. falciparum. P. malariae causes quartan malaria, an infection that is common in localized areas of the tropics.
P. ovale is the cause of a rare malarial infection with a periodicity like that of P. vivex, but it is milder and more rapidly cured4.
Anti malarial drugs are designed to prevent or cure malaria. Resistance to anti malarial drugs arises as a result of spontaneously-occurring mutations that affect the structure and activity at the molecular level of the drug target in the malaria parasite or affect the access of the drug to that target 5. Mutant parasites are selected if antimalarial drug concentrations are sufficient to inhibit multiplication. The emergence and rapid spread of P. falciparumresistance to commonly used antimalarial drugs poses a serious challenge to the effectiveness of early diagnosis and prompt treatment as a priority strategy within current malaria control efforts 6, 7.
The potential value of herbal malarial therapy using combinations of physiochemicalArtemether8 and Quinine9 was identified as a strategic and viable option in improving efficacy, and delaying development and selection of resistant parasites.
6.2 / Review of literature
The bark of the cinchonatree is used to make quinine. The medicinal properties of the cinchona tree were originally discovered by the Quechua Indians of Peru and Bolivia; later, the Jesuits were the first to bring the cinchona to Europe.Quinineis a natural white crystallinealkaloid having antipyretic, antimalarial, analgesic, and anti-inflammatory properties and a bitter taste. Quinine was the first effective treatment for malaria caused by Plasmodium falciparum, appearing in therapeutics in the 17th century. It remained the antimalarial drug of choice until the 1940s, when other drugs replaced it. Since then, many effective antimalarials have been introduced, although quinine is still used to treat the disease in certain critical situations10.
Artemisia annua is a common herb and has been found in many parts of the world11. Artemether is a lipid soluble methylether of dihydroartemisinin is a novel sesqiterpenelactone, extracted from the leaves of the shrub Artemisiaannua (annual wormwood) 12 and possesses an endoperoxide bridge which is a rare feather in natural products. The endoperoxide bridge is essential for its antimalarial activity 13. It is a white crystalline powder with a bitter taste. The drug is named Qinghaosu14 in Chinese. It was one of many candidates then tested by Chinese scientists from a list of nearly 200 traditional Chinese medicines for treating malaria. It was the only one that was effective, but it was found that it cleared malaria parasites from their bodies faster than any other drug in history.
Artemether have been used in combination with other antimalarial drugs for the treatment of malaria 15.
The concept of combination therapy of Artemether and Quinine based on the synergistic or additive potential of two or more drugs, to improve therapeutic efficacy and also delay the development of resistance to the individual components of the combination 16.
Combination therapy (CT) with antimalarial drugs is the simultaneous use of two or more blood schizontocidal drugs with independent modes of action and different biochemical targets in the parasite. Similarly, certain antimalarial drugs that fit the criteria of synergistic fixed-dose combinations are operationally considered as single products in that neither of the individual components would be given alone for antimalarial therapy.
Artemisinin-based combination therapies have been shown to improve treatment efficacy and also contain drug resistance in South-East Asia 17.
6.3 / Objectives of the study
The population at riskof malaria infection to have access to safe, good quality, effective, affordable and acceptable antimalarial. The objective of the proposal here associated to formulation.
Formulation of a combined antimalarial tablet dosage of Artemether and quinine.
Evaluation of combined antimalarial tablet dosage and study of stability of final product.
Analytical method Validation of antimalarial formulation by HPLC.
MATERIALS AND METHODS
7.1 / Sources of data
Wikipedia
Journal of Infectious Diseases
World Health organization
Science direct
World Wide Web
Pubmed
J-Gate@ Helinet etc
7.2 / Method of collection of Data
The sources of data are from the laboratory experiments, which involve formulation development and its validation.
7.3 / For formulation development
1. Formulation of combined Artemether and Quinine tablet formulation18.
2. Evaluation of tablets:
Hardness test
Friability test
Thickness test
Weight variation
Drug content assay
Dissolution study
Stability study of final selected formulation.
7.4 / For analytical method development :
The main objective of method development is to obtained a good separation with minimum time and efforts, based on goal of separation, the method development is proceed 19.
Recommended method development goals20:
Adequate resolution of all peaks, RS>2.0
Analysis time below 30 min, 20 min preferred.
Retention of first peak preferred to be at least K= 1.
Use buffered mobile phase and try low pH first.
Cost.
Minimum analysis time, Solvent consumption, waste.
Adequate column life time.
Easy to use and minimum training required.
7.5 / Analytical Method Validation of Artemether and Quinine by HPLC.
Unlike any other commodity, the manufactured herbal drug formulations must strictly comply with standard and regulations set forth by concerned authorities. The multicomponentherbal formulations are becoming popular in order to ensure timely and complete medication in multidrug therapy and enhancement of patient compliance. Analytical methods are extensively available for single drug formulations. Due to complexity in multicomponent formulations, method development poses a challenge for the analytical chemist. Official books also do not provide methods for simultaneous analysis. Most of the methods available for the analysis of active ingredients of such herbal formulations are applicable only after prior separation, which make it tedious and time consuming and also costly affair.
The objective of this validation study is to demonstrate that the proposed method is suitable for its intended use.
This validation study covers following parameters21, 22
a)Precision
- System precision
- Method precision
- Ruggedness
c) Linearity
d) Accuracy as recovery
e) Range
f) Solution stability
7.6 / DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS /ANIMALS? IF SO PLEASE DESCRIBE BRIEFLY.
No.
7.7 / HAS ETHICAL CLEARANCE BEEN OBTAINED FROM INSTITUTION IN CASE OF?
Not applicable.
8 / REFERENCES
1. Goodman & Gilman’s. The Pharmacological Basis of Therapeutics. 9th ed. New York: The McGraw Hill; 1996: 965
2. Malaria. Disease Impacts and Long-Run Income Differences (PDF). Institute for the Study of Labor 2007. URL:
3. Snow RW, Guerra CA, Noor AM, Myint HY, Hay SI. The global distribution of clinical episodes of Plasmodium falciparum malaria. Nature 2005; 434 (7030):214–47.
4. Plasmodium. Newsletter of the National Institute of Malaria Research, New Delhi. July 2009, 1(3). URL:
5. Poole S, editor. The use of antimalarial drugs: report of an informal consultation. Geneva: WHO; 2001: 7-8. URL:
- Bloland PB, Lackritz EM, Kazembe PN, Were JB, Steketee R, Camphell CC. Beyond chloroquine: implications of drug resistance for evaluating malaria therapy efficacy and treatment policy in Africa. The J of Infect Dis, 1993; 167(4):932–37.
- Bloland PB and Ettling M. Making malaria treatment policy in the face of
- Geyer M, editor. Antimalarial drug combination therapy: report of a WHO technical consultation. Geneva: WHO; 2001: 5-8. URL:
- Druilhe P, Brandicourt O, Chongsuphajaisiddhi T, Berthe J. Activity of a combination of three Cinchona bark alkaloids against Plasmodium falciparum in vitro. Antimicrob Agents and Chemother, 1998; 32(2): 250-54.
11. Peters W. The prevention of antimalarial drug resistance. Pharmacology
and Therapeutics, 1990; 47: 497-508.
12. URL:
13. Tripathi KD. Essentials of medical pharmacology. 4th ed. New Delhi:Jaypee
Brothers Medical Publishers (P) Ltd; 2003: 797-802
14. Trease and Evan’s. Pharmacognosy. 14th ed. London: W B Saunders
Company (P) Ltd; 2001: 426-9
15. Hien TT. An overview of the clinical use of artemisinin and its derivatives in
the treatment of falciparum malaria in Viet Nam. Transactions of the Royal
Society of Tropical Medicine and Hygiene, 1994; 88(1): 7-8.
16.White NJ. Delaying antimalarial drug resistance with combination Chemotherapy.Parassitologia. 1999; 41: 301-8.
17. White NJ and Olliaro PL. Strategies for prevention of antimalarial drug resistance: rationale for combination therapy for malaria. Parasitol Today.1996;12(10): 399-401.
18. Mark Gibson. Pharmaceutical Preformulation and Formulation. A practical guide from candidate drug selection to commercial dosage form.Baeo Raton: Taylor and Francis gp; 2001: 379-403
19. Sethi PD. High Performance Liquid chromatography, Quantitative analysis of pharmaceutical formulations; 1st ed.New Delhi:CBS Publisherand Distributors;2001: 5- 201
20. Lloyd R. Snyder, Joseph J. Kirkland, Joseph L, Glajch; Practical HPLC method development; 2nd ed.INC: John Wiley and Sons;1997: 233-89
21. Quanyun A, Xu Lawrence AT. Stability- indicating HPLC methods for drug analysis; 3rd ed. London: Pharmaceutical Press;2008: 52, 72722.
22. Ermer J, John H, Miller McB. Method validation in pharmaceutical Analysis. KGaA: Wiley-VCH verlagGmbh and Co; 2005: 17-170
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/SIGNATURE OF THE CANDIDATE
/ (Mr. S. S Kushwaha.)10 / REMARKS OF THE GUIDE / The above mentioned information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.
11 / NAME AND DESIGNATION OF GUIDE
SIGNATURE
/ Mr. B. S. PATIL,M. Pharm, (Ph.D)Asst. Professor,,
Post Graduate Department of Pharmacognosy,
SET’s College of Pharmacy, S.R.Nagar,
Dharwad. Karnataka – 580 002.
12
/NAME AND DESIGNATION OF CO – GUIDE
SIGNATURE / ------13 / NAME AND DESIGNATION OF HOD
SIGNATURE / Dr. P. V. HABBU,M. Pharm, Ph.D.
Professor & HOD
Post Graduate Department of Pharmacognosy,
SET’s College of Pharmacy, S.R.Nagar,
Dharwad. Karnataka – 580 002.
14
/REMARKS OF PRINCIPAL
/ The above mentioned information is correct and I recommend the same for approval.15 / NAME OF THE PRINCIPAL
SIGNATURE / Dr. V. H. KULKARNI, M. Pharm, Ph.D.
Principal, SET’s College of Pharmacy,
S. R. Nagar, Dharwad. Karnataka – 580 002.