Development of New Analytical Methods for Estimation of Some Hemostatic Drugs

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. Name of the Candidate and M.D ARSHAD Address (in block letters) H.NO: 5-993/2 B MEHBOOB NAGAR COLONY ROZA KHURD GULBARGA, KARNATAKA. 2. Name of the Institution H.K.E.S’S COLLEGE OF PHARMACY, SEDAM ROAD, GULBARGA-585105 (KARNATAKA) 3. Course of Study and Subject M. PHARM. (PHARMACEUTICAL ANALYSIS)

4. Date of Admission to Course 20-6-2009

5. Title of the Topic “DEVELOPMENT OF NEW ANALYTICAL METHODS FOR ESTIMATION OF SOME HEMOSTATIC DRUGS’’ 6. BRIEF RESUME OF THE INTENDED WORK 6.1 Need for the Study:

Ethamsylate is an orally available hemostatic drug. It is official in British Pharmacopoeia. Investigations of some new economical analytical methods are in need for the quantitative estimation of Ethamsylate in bulk drug and pharmaceutical dosage forms with high sensitivity, accuracy, precision.

6.2 Review of Literature:

Ethamsylate1-3is chemically Diethylammonium 2,5-dihydroxybenzenesulphonate.

Molecular formula for Ethamsylate is C10H17NO5S, Molecular weight is 263.31, Melting point is 125C, soluble in water and alcohol. Ethamsylate4 has been used in the prevention and treatment of capillary bleeding in menorrhagia after abortion, epistaxis, malena, hematuria and after tooth extraction but efficacy is unsubstantiated. There are few reported methods available for determination of Ethamsylate by RP- HPLC5 ,and HPTLC6 method for the determination of Ethamsylate in bulk and tablet dosage. O O S OH HO H . H3C N CH3 OH Structure of Ethamsylate ( I )

6.3 Objectives of the Study:

O O S OH HO R =

Since few analytical methods have been reported for the quantitative estimation of Ethamsylate, there is a necessity for investigation of new analytical methods for quantitative estimation of Ethamsylate in bulk drug and pharmaceutical dosage forms (Tablets).In view of the above facts, the following analytical methods are planned to develop

I) Since the drug Ethamsylate is sufficiently soluble in water and ethanol, 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 hydroxy group,it can form colored chromogens with phosphomolybdotungstic acid, well known as Folin - ciocalteu reagent ( FC reagent ) in alkaline pH by redox reaction which can be used for quantitative estimation of Ethamsylate in bulk drug and pharmaceutical dosage forms by visible spectrophotometry. III) Presence of phenolic hydroxyl group in Ethamsylate allows the oxidative coupling reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) (II) in presence of Cerric ammonium sulphate forms colored chromogen(III) by which drug can be estimated quantitatively by spectrophotometrically.

N N - NH2 + R - OH

CH3

( II )

Cerric ammonium sulphate

+ N N = N - R - OH

CH3 Colored chromogen ( III )

IV) The phenolic hydroxy group in drug molecule reacts with ferric salt (ferric chloride) in aqueous, alcoholic or chloroform media to give intense coloration characteristic of the drug. The color is due to the strongly ionized complex, phenolate of trivalent iron. The colored chromogen (IV) formed can be utilized for quantitative estimation of drug by colorimetrically.

+ - 3- FeCl3 + 6 ROH 6H + 3Cl + [ Fe(OR)6]

(I) (IV) Colored chromogen

V) Since the drug is having phenolic group, it form colored complex (V, VI) with 1,10-phenanthroline and 2,2l-bipyridine in presence of Fe+3 which can be Utilized for quantitative estimation of Ethamsylate in bulk drug and pharmaceutical dosage forms colorimetrically.

R - OH + Fe+3 ( I )

Fe+2

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

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 Ethamsylate in bulk drug and pharmaceutical dosage forms with high sensitivity, accuracy, precision and which are economical.

7. MATERIALS AND METHODS:

In the present work investigation of new analytical methods for quantitative estimation of Ethamsylate, 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 obtained by Dr. Reddy’s laboratory, Hyderabad. 7.1 Source of Data:

a. Internet, 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.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 Ethamsylate will be obtained from Dr. Reddy’s laboratory, Hyderabad.

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 and Biologicals,14th edn, Merck & Co. Inc: 2006; 3727.

2. Sweetman SC, editor. Martindale: The Complete Drug Reference, 35th edn Pharmaceutical Press, London: 2007; 959.

3. British Pharmacopoeia. Medicines and Health care products regulatory agency (MHRA), London. 2008; 2: 840.

4. Tripathi.K.D,Essentials of Medical Pharmacology, 6th edn 2008;596.

5. P. Nagaraju*,K. Krishnachaithanya,VDN.Srinivas and SVN. Padma. RP-HPLC method for the determination of Ethamsylate in bulk and tablets dosage forms. Asian journal research chem.2008;1(2):0974-4169.

1 1 1 6. Yogini S. Jaiswal ,Gokul S. Talele ,Sanjay J. Surana A Simple and sensitive HPTLC method for quantitative analysis of Ethamsylate in tablets. journal of planar chromatography-modern TLC.2007;18:380-383.

9. Signature of Candidate

10. Remarks of the Guide The work under taken is novel and can be considered for quality control in a routine manner.

Name & Designation of (in block letters)

11. 11.1 Guide Mr. ARVIND.B.KARADI,M.Pharm(Ph D) ASSISTANT PROFESSOR DEPT. OF PHARMACEUTICAL ANALYSIS, H.K.E.S’S COLLEGE OF PHARMACY, GULBARGA

11.2 Signature

11.3 Head of the Department DR. S. M. MALIPATIL, M.Pharm,Ph.D PROFESSOR DEPT. OF PHARMACEUTICAL ANALYSIS, H.K.E.S’S COLLEGE OF PHARMACY, GULBARGA

11.4 Signature

12. 12.1 Remarks of the Chairman & principal 12.2 Signature