DEVELOPMENT OF VALIDATED ANALYTICAL METHODS

FOR THE ESTIMATION OF SOME NEW ANTICOAGULANT

DRUGS IN PHARMACEUTICAL DOSAGE FORMS

M. Pharm Dissertation Protocol

Submitted to

RajivGandhiUniversity Of Health Sciences,
Bangalore,Karnataka

By

p.digamber patel

Under the guidance of

Dr.S.APPALA RAJU

M.Pharm., Ph.D.

DEPARTMENT OF PHARMACEUTICAL ANALYSIS

H.K.E.S’S MATOSHREE TARADEVI RAMPURE INSTITUTE OF PHARMACEUTICAL SCIENCES,

GULBARGA – 585105

2011-12

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES KARNATAKA, BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the Candidate and
Address (in block letters) / P.DIGAMBER
S/OP.ANANDRAO
H.NO:5-16,BASAR(VILL)
MUDHOLL(MDL)
DIST:ADILABAD
ANDHRA PRADESH-504101
2. / Name of the Institution / MATOSHREE TARADEVI RAMPURE INSTITUTEOF PHARMACEUTICAL SCIENCES, Sedam Road, GULBARGA-585105.
3. / Course of Study and Subject / M. Pharm.
(PHARMACEUTICAL ANALYSIS)
4. / Date of Admission to Course / 18/12/2011
5. / Title of the Research topic / “DEVELOPMENT OF VALIDATED ANALYTICAL METHODS FOR THE ESTIMATION OF SOME NEW ANTICOAGULANT DRUGS IN PHARMACEUTICAL DOSAGE FORMS”
6. / Brief Resume of Intended Work
6.1 Need for study
Acenocoumarol is an oral anticoagulant of the coumarin family.It is the most widely prescribed drug in deep venous thrombosis pulmonary embolism and arterial thromboembolism in patients with atrial fibrillation.Hence it is used as an anticoagulant agent for the management of thromboembolic disorders. There is wide scope for the development of new analytical methods for the assays of Acenocoumarol.Development of some new instrumental methods are in need for the quantitative estimation of Acenocoumarol in bulk drug and pharmaceutical dosage forms with high sensitivity,accuracy,precision and economical too.
6.2 Review of Literature
Acenocoumarol1 is chemically (RS)-4-hydroxy-3-(1,4-nitro phenyl-3-oxobutyl)coumarin.The molecular formula is C19H15NO6,Molecular weight is 353.3 and melting point is 196-1990.It is practically insoluble in water and in ether,slightly soluble in alcoholand in chloroform.It dissolves in aqueoussolutions of alkali hydroxide2,3.It is official in Indian and British pharmacopoeia.Literature survey reveals that:
1)A.S.Mehta et al developed aspectrophotometric methods for estimation of Acenocoumarol in bulk and its pharmaceutical dosage forms4.
2)B.M.Gurupadayya et al developed visible spectrophotometric methods for the estimation of Acenocoumarol in tablets5.
3)R.Ceresole et al developed a HPLC method for determination of Acenocoucomarol and its major thermaldegradation product6.
4)I.J.Terpstra et al developed a placebo-controlled pharmacodynamic and pharmacokinetic interaction study between tamsulosin and acenocoumarol7.
5) E.Moore and C Lau-Cam developed liquid chromatographic determination of coumarin anticoagulant in tablets8.
6)K.M.Rentsch et al developed asensitive stereospecific determination of acenocoumarol and phenprocoumon in plasma by HPLC9.
The analytical important functional Hydroxyl and Nitro group in Acenocoumarol have not been exploited completely.Hence there is ascope for developing more spectrophotometric methods for Acenocoumarol.The Structural formula of Acenocoumarol is as shown.

6.3 Objectives of the Study:
Many new analytical methods can be investigated for the Quantitative estimation of Acenocoumarol in bulk drug and dosage forms.Thus,the following analytical methods are planned to develop with high sensitivity,accuracy and precision.
1.Since the drug Acenocoumarol is soluble in methanol,ethanol and aqueous solutions of alkali,UV-Spectrophotometric methods can be developed for its quantitative estimation in bulk drug and pharmaceutical dosage forms.
2.The analytical important hydroxyl group in Acenocoumarol can be completely exploited by reacting the drug with suitable analytical reagents like Folin’s Ciocalteu reagent,MBTH(3-methyl-2-benzothiazolinone hydrazone), 2,2-Bipyridine,1,10-phenanthroline,Gibb’s reagent(2,6-dicholroquinone-4-chlorimide) to form coloured chromogens by which acenocoumarol can be estimated in bulk drug and pharmaceutical dosage forms by visible spectrophotometry.
3. The analytical important nitro group in acenocoumarol after reduction can be reacted with suitable analytical reagents like ninhydrin reagent in DMF, ascorbic acid in DMF, p-benzoquinone (PBQ) to give coloured condensation products .
The reduced acenocoumarol can also undergo diazotization coupling reaction with Bratton Marshall reagent to give coloured chromogens by which acenocoumarol can be estimated in bulk drug and pharmaceutical dosage forms by visible spectrophotometry.
i)Reduced acenocoumarol reacts with ninhydrin reagent in DMF which results in the
formation of diketohydrindylindene-diketohydrindamine (Ruhemenn’s purple) which can be utilized for the quantitative estimation in bulk drug and pharmaceutical dosage forms.


ii)The amino group in reduced acenocoumarol reacts with ascorbic acid in DMF medium to produce a coloured product. Ascorbic acid undergoes oxidation resulting in the formation of dihydro ascorbicacid. The carbonyl group of dihydroascorbate reacts with the –NH2 group of reduced acenocoumarol to form a coloured condensation product which can be utilized for the quantitative estimation of the drug.

iii)The amino group in reduced acenocoumarol reacts with p-benzoquinone(PBQ) to give coloured product. The amino group condenses with carbonyl group of PBQ to form coloured condensation product which can be utilized for the quantitative estimation of the drug.

iv)The amino group in reduced acenocoumarol undergoes diazotization followed by coupling with Bratton Marshall reagent [N-(1-naphthyl)-ethylenediamine dihydrochloride] to give coloured chromogen which can be utilized for the quantitative estimation of the drug .

4. Reverse phase HPLC method can also be developed using (Shimadzu HPLC class VP series 6.01) with two LC-10 ATVP pumps variable wavelength programmable uv-visible detector for quantitative estimation of acenocoumarol in bulk drug and pharmaceutical dosage forms with high accuracy, sensitivity and precision.
5. Liquid chromatography with mass spectrometric method can also be developed for quantitative estimation of acenocoumarol in bulk drug and pharmaceutical dosage forms.
7.Material and Methods
In the present investigation of new analytical methods for quantitative estimation ofAcenocoumarol, 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 is collected fromAbbott Healthcare Pvt.Ltd, Mumbai– 400 080.
7.1 Source of Data

1.Internet, Library

  1. GulbargaUniversity, Gulbarga
  2. I.I.Sc. Library, Bangalore
  3. I.I.C.T. Library, Hyderabad
  4. R.G.U.H.S. Library, Bangalore
7.2 Methods of Collection of Data (including sampling procedure, if any)

Data collected from

  1. Internet, H.K.E.S. College of Pharmacy, Gulbarga.
  2. Analytical abstract and chemical abstract GulbargaUniversity, I.I.C.T. & I.I.SC. Libraries.
  3. Journals like – Indian J. Pharmaceutical Sciences, Indian Drug & Indian J. Analytical Chemistry.
  4. e-Journals
  5. Drug sample of Acenocoumarol is collected from Abbott healthcare pvt.Ltd, Mumbai– 400 080.
7.3 Does study require any investigation 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-

References
  1. O’Neil M.J, editor, The Merck Index An Encyclopedia of Chemicals, Drugs and Biologicals, Merck & Co. Inc. 14th edition, 2006: 07.
  1. Sweetman SC, editor, Martindale The Complete Drug Reference, Pharmaceutical Press, London (U.K), 35th edition,2007:1078.
  1. CIMS-115,UBM medica Drug Reference world wide,Oct.11-Jan.12,17.
  1. MehtaA.S.,GurupadayyaB.M.,GopinathB.,Manohara Y.N.and Akhilesh Chandra, Int.J.Chem.Sci.: 2008,6(2),1067-1073.
  1. Ankita S.M,Gurupadayya B.M, Gopinath B, Manohara Y.N and Akhilesh Chandra,Indian J.Pharm.Educ.Res; 2008,42(4),310-313.
  1. Ceresole R, Rosasco M.A, Forastieri C.C & Segall A.I, Jour.Of. Liq.Chroma. & Related Techno., 2007;31(2).
  2. Rolan P, Terpstra I.J, Clarke C, Mullins F, & Visser J.N, Br.J.Clin.Pharmacol ,2003;55(3): 314-316.
  3. MooreE, Lau-Cam C, J.Assoc.Off Anal Chem, 1986; 69(4):629-32.
  1. Rentsch KM, Gutteck-Amsler U, Buhrer R, Fattinger KE, Vonderschmitt DJ, J.Chromatogr B Biomed Sci Appl, 2000; 742(1): 131-42.

9. / Signature of candidate
10. / Remarks of Guide / The work undertaken is novel and results oriented
11. / Name and designation of
(in block letters) / Dr. S. Appala Raju
Principal
11.1 Guide / Dr.S.APPALARAJU
M.Pharm., Ph.D.
PROFESSOR
DEPT. OF PHARMACEUTICAL ANALYSIS,MATOSHREE TARADEVI RAMPURE INSTITUTE OF PHARMACEUTICAL SCIENCES,
GULBARGA
11.2 Signature
11.3 Co-guide /
Dr. SHOBHA MANJUNATH M.Sc., Ph.D.
PROFESSOR
DEPT. OF PHARMACEUTICAL ANALYSIS,MATOSHREE TARADEVI RAMPURE INSTITUTE OF PHARMACEUTICAL SCIENCES,
GULBARGA
11.4 Signature
11.5 Head of the department / Dr. S.M.MALIPATIL
M.Pharm., Ph.D.
PROFESSOR
DEPT. OF PHARMACEUTICAL ANALYSIS, MATOSHREE TARADEVI RAMPURE INSTITUTE OF PHARMACEUTICAL SCIENCES,
GULBARGA.
11.6 Signature
12 / 12.1 Remarks of Principal
12.2 Signature