“Development of New analytical Methods for estimation of some antiviral drugs”

M. Pharm Dissertation Protocol

Submitted to the

Rajiv Gandhi University Of Health Sciences, Karnataka, Bangalore

By

MISS. JADHAV SUVARNA VYANKATRAO

Under the guidance of

Dr. S. APPALA RAJU

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 and
Address (in block letters) / Miss. Jadhav Suvarna Vyankatrao
H.K.E.S’S College of Pharmacy, Sedam Road, GULBARGA-585 105 (Karnataka)
2. / Name of the Institution / H.K.E.S’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 / 24.06.2008
5. / Title of the Topic / “DEVELOPMENT OF NEW ANALYTICAL METHODS FOR ESTIMATION OF SOME ANTIVIRAL DRUGs”
6. / BRIEF RESUME OF THE INTENDED WORK
6.1 Need for the Study
Nelfinavir mesilate is an antiviral drug which bind to protease molecule, interfere with its cleaving function and inhibit it. It is not official in any pharmacopoeia. Investigation of some instrumental methods are in need for the quantitative estimation of nelfinavir mesilate in buk drug and pharmaceutical dosage forms with high sensitivity, accuracy, precision and economical too.

6.2Review of Literature

Nelfinavir mesilate1,4 (I) is chemically (3S, 4aS, 8aS)- N-(1,1-Dimethylethyl)decahydro-2-[(2R,3R)-2-hydroxy-3-[(3-hydroxy-2-methyl benzoyl)amino]-4-(phenylthio) butyl]-3-isoquinoline carboxamide used as an antiviral agent which is a retroviral protease inhibitor5-8. It is not official in any pharmacopiea. It occur as white poeder which is nonhygroscopic. It is very soluble in methanol, ethanol, acetonitile but practically insoluble in water, soyabean oil, mineral oil9. Its molecular formula is C32H45N3O4.CH3SO3, its molecular weight is 663.89 g.

Literature survey revealed that one spectrophotmetric method has been published on nelfianvir mesilate10.

( I )
6.3 Objectives of the Study
i) Since the drug nelfinavir mesilate is sufficiently soluble in methanol, ethanol and acetonitrile a number of UV spectrophotometric methods can be developed for its quantitative estimation in bulk drug and pharmaceutical dosage forms.
ii)Since the drug is having phenolic group, it can form coloured chromogen with phosphomolybdo tungstic acid, well known as Folin-Ciocalteu reagent11 (FC reagent) in alkaline pH by redox reaction which can be used for quantitative estimation of nelfinavir mesilate in bulk drug and pharmaceutical dosage forms by visible spectrophotometry.
iii)Presence of phenolic group in nelfinavir mesilate allows the oxidative coupling reaction with 3-methyl-2-benzothiazolinone hydrazone12 (MBTH) (II) in presence of ceric ammonium sulphate and forms coloured chromogen (III) by which drug can be estimated quantitatively by visible spectrophotometry.
iv) / The phenolic hydroxy group in drug molecule reacts with ferric salt (ferric chloride) in aqueous, alcholic or chloroform media to give intense colouration characteristic of the drug. The colour is due to the strongly ionised complex, phenolate of trivalent iron13. The coloured chromogen (IV) formed can be utilised for quantitative estimation of drug by visible spectrophotometry.

7. / FeCl3 + 6 ROH6H+ + 3Cl- + [ Fe(OR)6]3-
(I) (IV)

Since drug is having phenolic group, it forms coloured chromogen (VI) with Gibb’s reagent (2,6-dichloroquinone chlorimide (V) in alkaline pH which can be utilised for quantitative estimation of drug by visible spectrophotometry in bulk drug and pharmaceutical dosage forms.

(VI)
/ vi) Since the drug is having phenolic group, it form coloured complexes (VII, VIII) with 1,10-phenanthroline and 2,2’-bipyridine in presence of Fe (III) which can be utilized for quantitative estimation of nelfinavir in bulk drug and pharmaceutical dosage forms colorimetrically14-15.
vii) Reversed phase high performance liquid chromatographic techniques can be developed using a gradient HPLC (Shimadzu HPLC Class VP series 6.01) with two LC-10 AT VP Pumps, variable wave length programmable UV/visible detector for quantitative estimation of nelfinavir mesilate in bulk drug and pharmaceutical dosage forms with high sensitivity, accuracy, precision and which are economical too16.
7. / MATERIALS AND METHODS
In the present investigation of the new instrumental methods for quantitative estimation of Nelfinavir mesilate, we are in need and using Shimadzu 1700 double beam UV/visible spectrophotometer, HPLC (Shimadzu, Class VP series 6.01), chromatographic instruments and volumetric glass apparatus. Drug sample will be provided by Cipla Ltd, Mumbai.
7.1 Source of Data
a. Internet, Gulbarga University Library
b. Gulbarga University, Gulbarga
c. I.I.Sc. Library, Bangalore
d. I.I.C.T. Library, Hyderabad
e. R.G.U.H.S. Library, Bangalore
7.2 / Methods of collection of data (including sampling procedures, if any)
Data Collected From
i)Internet: H.K.E.S’s College of Pharmacy, Gulbarga.
ii)Analytical Abstracts and Chemical Abstracts – Gulbarga University, I.I.Sc. 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 nelfinavir mesilate will be collected from Cipla Ltd, Mumbai
7.3 / Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If so, please describe briefly.
---- No ----
7.4 / Has ethical clearance been obtained from your institution in case of 7.3.
---- Not Applicable ----
8. / References:
  1. O’ Neil MJ, editor, The Merck Index: An Encyclopedia of Chemicals, Drug, Biologicals, 14th edn. Merck & Co. Inc.: 2006: p. 1114.
  2. Sweetman SC, editor, Martindale: The Complete Drug Reference, 35th edn. Pharmaceutical Press, London: 2007: p. 804.
  3. Bertam G Katzung, editor, Basic and Clinical Pharmacology, 9th edn. Mc. Graw Hill, singapore, 2007: p.870.
  4. CIMS, Multimedia Health Pvt. Ltd., Bangalore, 2008: p. 401
  5. Patrick AK, et al. Genotypic and phenotypic characterization of human immune efficiency virus type 1 variants isolated from patients treated with the protease inhibitor nelfinavir, Antimicrob Agents chemother, 1998; 42: 2637-44.
  6. Pai VB, Nahata MC, Nelfinavir mesylate: a protease inhibitor, Ann Pharmacother, 1999; 33: 325-39.
  7. Perry CM, et al. Nelfinavir: a review of its use in the management of HIV infection. Drugs 2005; 65: 2209-44.
  8. Shetty BV, et al. Preclinical Pharmacokinetics and tissue distribution, Ibid, 110.
  9. Longer M. et al. Nelfinavir: Physical properties, J. Pharm. Sci., 1995; 84: 1090.
  10. Mohan Rao SVM, Subba Reddy TR, Colorimetric methods of estimation of nelfinavir mesylate, Indian Drugs 2003, 40: 467-70.
  11. Ramana Rao, Kanjilal G, Mohan KR. Analyst 1978, 103, 993.
  12. Sawicki E, Hauser TR, Stanely TW, Elbert W. Anal. Chem, 1961, 93,33.
  13. Morrison RT, Boyd RN. Editor, Organic Chemistry, 6th edn., John Wiley and sons, Inc. New York, p. 197,923.
  14. Chowdhary KRR, Rao GD. Indian Drugs, 1997:34(5):283.
  15. Besada A, Anal. Lett. 20:427
  16. Heftmann E, editor, Chromatography, 1992, Elsevier, Amsterdam.: 112

9. / Signature of Candidate / SUVARNA V. JADHAV
10. / Remarks of the Guide / The work undertaken is novel and results oriented that may lead to new research findings.
11. / Name & Designation of (in block letters)
11.1Guide / Dr. S. AppAla RajuM. Pharm., Ph.D
Principal and Professor
Dept. of Pharmacutical Analysis
H.K.E.S’s College of Pharmacy,
Gulbarga-585105
(Karnataka)
Signature
11.2 Co-guide / Dr. Shobha Manjunath
M.Sc. Ph. D
Professor
Dept. of Pharmaceutical Analysis
H.K.E.S’s College of Pharmacy,
Gulbarga-585105
Signature
11.3Head of the Department / Dr. S. M. MalipatilM.Pharm., Ph. D
Professor
Dept. of Pharmaceutical Analysis
H.K.E.S’s College of Pharmacy,
Gulbarga-585105
Signature
11.4Remarks of the Chairman & Principal
Signature