“Development of new analytical methods for Quantitative estimation of Antiviral agents”
M. Pharm Dissertation Protocol
Submitted to the
Rajiv Gandhi University Of Health Sciences, Karnataka, Bangalore
By
karazgi kishwar jahan
Under the guidance of
Dr. S.M. Malipatil
M.Pharm., Ph.D
DEPARTMENT OF pharmaceutical analysis
H.K.E’S COLLEGE OF PHARMACY,
GULBARGA – 585105
2008-09
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA
BANGALORE
ANNEXURE - II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidate & Address(In block letters) / : / KARAZGI KISHWAR JAHAN
H-NO 13-55 NEAR MASJID E AMEENA
ISLAMABAD COLONY GULBARGA-585104.
2. / Name of the Institution / : / H.K.E. Society’s College of Pharmacy, Sedam Road,Gulbarga -585 105. (kARNATAKA)
3. / Course of Study and Subject / : / M.Pharm
(Pharmaceutical ANALYSIS)
4. / Date of Admission to Course / : / 04.06.2008
5. / Title of the Topic / : / DEVELOPMENT OF NEW ANALYTICAL METHODS FOR QUANTITATIVE ESTIMATION OF ANTIVIRAL AGENTS
6. /
Brief resume of the intended work
6.1 / Need for the study:Oseltamivir has antiviral activity similar to that of zanimivir Its active metabolite, Oseltamivir Carboxylate, selectively blocks the viral surface enzyme neuraminidase, thereby preventing the release of virus particles from infected cells. Oseltamivir is active against influenza A and B virus neuraminidase.
Oseltamivir is not official in any pharmacopoeia. Investigations of new analytical methods are in need for the quantitative estimation of Oseltamivir in bulk as well as pharmaceutical formulations with high sensitivity accuracy, precision and economy.
6.2 / Review of Literature:
Oseltamivir1 is chemically (3R,4R,5S)-4-(Acetylamino)-5-amino-3-(1-ethyl propoxy)-1-cyclohexene-1-carboxylic acid ethyl ester. The molecular formula is C16H28N2O4 and molecular weight is 312.40.C 61.51%, H9.03%, N 8.97%, O 20.49%. Orally active inhibitor of influenza virus neuraminidase; converted in-vivo to the active acid metabolite. Neuraminidase inhibitors2 are effective against all strains of influenza and de novo resistance3 has not been found to date. Some studies4,5 have been described in-vitro resistance to Oseltamivir arising during clinical use but it is not yet known if such resistant viruses are transmissible and pathogenic. No resistant viruses have been isolated from immunocompetent people given zanamivir6.
A literature survey reveals that there are few published reports of HPLC methods7 for determination of Oseltamivir, and a sensitive HPLC-Mass spectrometric8,9 assay for Oseltamivir carboxylate in plasma and urine.
Chemical structure of Oseltamivir:
6.3 / Objectives of the Study:
The aim of the present work is to develop some simple, accurate, precise and economical analytical methods for the quantitative estimation of Oseltamivir from bulk drugs and pharmaceutical dosage forms.
Since the literature survey reveals that only HPLC methods for Oseltamivir Phosphate in pharmaceutical preparations have been described, and a sensitve HPLC-Mass spectrometry assay for Oseltamivir Carbooxylate in plasma and urine has been developed.
In view of the above facts the following analytical methods are planned to develop with high sensitivity, accuracy and precision.
(i) Since Oseltamivir (I) is having an amino group it can be condensed with various aromatic aldehydes (II) like P-dimethylaminobenzaldehyde(PDAB), P-dimethylaminocinnamaldehyde (PDACA) and vanillin to get coloured Schiff’s bases (III) and can be used for quantitative estimation of drug by visible spectrophotometer.
/ R – NH2 + OHC – R’
(I) (II)
R – N = CH – R’
(III)
Coloured chromogen or Schiff’s base
ii) Amino group can be diazotized with nitrous acid (HNO2) and coupled with chromogenic agents (IV) like N-(1-napthyl) ethylendiamine dihydrochloride , β-napthol , 2-napthalamine and phloroglucinol to get coloured chromogen (V) and drug can be estimated quantitatively by colourimetry.
Along with Brotton-Marshall Reagent, we can also use other coupling agents mentioned above and develop number of new analytical methods.
iii) Presence of amino group in Oseltamivir allows the oxidative coupling reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) in presence of ferric chloride or ceric ammonium sulphate and forms coloured chromogen (VI) by which drug can be estimated quantitatively by colourimetry.
Coloured chromogen
iv) Oseltamivir forms coloured complexes with1,10-phenanthroline and 2,2’-bipyridine in presence of Fe (III) which can be utilized for quantitative estimation of Oseltamivir in pharmaceutical formulation.
/ R – NH2 + Fe3+
Fe2+
1, 10-phenanthroline 2, 2’-Bipyridine
O-phosphoric acid O-phosphoric acid
Coloured chromogen Coloured chromogen
vi) Visible spectrophotometric methods can also be developed using chromogenic reagents like Gibb’s reagent and Folin-wu reagent due to presence of primary & secondary amino group in the drug.
vii) Simultaneous spectrophotometric methods and RP-HPLC can also be developed.
viii) Reversed phase HPLC methods can be developed by varying the conditions for estimation of bulk drugs and pharmaceutical formulation.
ix) Various HPTLC methods can also be developed.
7. / MATERIAL AND METHODS
In the present investigation of the new analytical methods for quantitative estimation of Oseltamivir, we are using Shimadzu 1700 double beam UV/ visible spectrophotometer, Shimazdu Prominence Isocratic HPLC system with LC 20 AT Pump, SPD-20A Detector, Spinchrome CFR Software, chromatographic instruments and volumetric glass apparatus. Drug sample is provided by Hetero Drugs (P) Ltd, Erragadda Hyderabad as gift sample for research. All other chemical reagents are from Qualigens, Mumbai.
7.1 / Sources of Data:
a) Internet
b) I.I.C.T. Library, Hyderabad.
c) I.I.S.C. Library, Bangalore.
d) RGUHS Library, Bangalore.
e) Gulbarga University Library, Gulbarga.
7.2 / Methods of collection of data (including sampling procedures, if any):
i) Internet: H.K.E.’s College of Pharmacy, Gulbarga.
ii) Analytical abstracts and chemical abstracts – Gulbarga University, I.I.S.C. and I.I.C.T. Libraries.
iii) Journals like – Indian J. Pharmaceutical Sciences, Indian Drugs and Indian J. Analytical Chemistry.
iv) E-Journals.
v) Drug sample of Oseltamivir is collected from Hetero Drugs (P) Ltd, Erragadda Hyderabad
7.3 / Does the study require any investigation or invervention to be conducted on patients or other humans or animals? If so please describe briefly.
------Not under the plan of the work ------
7.4 / Has ethical clearance been obtained from your institution in case of 7.3
------Not applicable------
8. / List of References:
1 / O’Neil, M.J. (Ed. By) (2006), The Merck Index – an encyclopedia of Chemicals, Drugs and Biologicals, Merck and Co., Inc, 14th Edition; 1187-1188.
2 / Sweetman, S.C. (Ed. By), (2007), Martindale – The Complete Drug Reference, Pharmaceutical Press, London (U.K), 35th Edition Vol (1); 806.
3 / Mckimm-Breschkin J, et,al , Anti microb Agents chemother 2003; 47: 2264-72.
4 / Whitney RJ, Et al. Pediatr Infct-Dist J 2001; 20: 127-33
5 / Kiso M, et al , Lancet 2004; 364: 759-65.
6 / Moscona A. Lancet 2004: 364: 733-34
7 / Michael D Green, Henry Netty and Robert A. Wirtz (2008) J.Emg Inf Dis, 14: 4
8 / Wiltshire H, Wiltshire B, Citron A, Clarke T, Serpe C, Gray D et al. J. Chromatgr B. 2000; 745:373-88
9 / Lindergardh N, Hien, Farrar J, Singhasivanan P, White NJ, Day J. Pharm Biomed Anal 2006; 42: 430-433.
9. / Signature of Candidate / (karAzgi kishwar jahan)
10. / Remarks of the Guide / The work undertaken is novel. It involves high precision and may results into new research finding.
11. / Name & Designation of (in block letters)
11.1 / Guide / Dr. S.M. MALIPATIL
M.Pharm., Ph.D.
PROFESSOR
Dept. of pharmaceutical analysis
H.K.E.S College of Pharmacy, gulbarga
11.2 / Signature
11.3 / Head of Department / Dr. S.M. MALIPATIL
M.Pharm., Ph.D.
PROFESSOR
Dept. of pharmaceutical analysis
H.K.E.S College of Pharmacy, gulbarga
11.4 / Signature
12. / 12.1 / Remarks of the Chairman & Principal
12.2 / Signature
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