Rajiv Gandhi University of Health Sciences s25

“STABILITY INDICATING INSTRUMENTAL METHOD FOR THE DETERMINATION OF AMPICILLIN AND SULBACTUM IN PHARMACEUTICAL DOSAGE FORMS.”

SYNOPSIS FOR M.PHARM DISSERTATION

SUBMITTED TO RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES KARNATAKA

BY SIDDALINGA PRASAD N. C. Ist. M. PHARM DEPARTMENT OF PHARMACEUTICAL ANALYSIS PES COLLEGE OF PHARMACY

BANGALORE-560050 (2007-08)

1 RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1 Name of the candidate and address Mr. SIDDALINGA PRASAD N. C. Department of Pharmaceutical Analysis, P.E.S.College of Pharmacy, 50 Feet Road, Hanumanthanagar, BSK Ist Stage, Bangalore-560050.

Permanent Address S/o Channabasava Raju N. G. Sri Gangadhareshvara Nilaya Sri Hippe Anjaneya Swamy Temple Layout. Adepete. Nelamangala. Bangalore Rural dist, Karnataka-563137.

2 Name of the institution P.E.S.College of Pharmacy 50 Feet Road, Hanumanthanagar, BSK Ist Stage, Bangalore-560050.

3 Course of study and subject MASTER OF PHARMACY IN PHARMACEUTICAL ANALYSIS

4 Date of the admission 10th June 2007

5 Title of the topic:

“STABILITY INDICATING INSTRUMENTAL METHOD FOR THE DETERMINATION OF AMPICILLIN AND SULBACTUM IN PHARMACEUTICAL DOSAGE FORMS.”

2 6 Brief resume of the intended work: 6.1 Need for the study: - Stability testing is a routine procedure performed on drug substances and products and it is involved at various stages of product development. In early stages, accelerated stability testing (at relatively high temperature and/or humidity) can be used for evaluation of drugs to determine what types of degradation products may be found after long-term storage. Stability testing is performed to ensure that drug products retain their therapeutic effectiveness until the end of their expiration dates. ICH guidelines recommended stability for active substances and related finished products. The purpose of stability testing is to provide evidence on how the quality of the active substances or finished product varies with time under the influence of variety of environmental factors such as temperature, humidity and light, and to establish a re-test period of the active substance for the shelf life for the finished product and recommend the storage conditions established. Stability of a pharmaceutical product may be defined as the capability of a particular formulation in a specific container/closure system to remain within its physical, chemical, microbiological, therapeutic, toxicological, protective and informational specifications, although there are exceptions, 90% of a labeled potency is generally recognized as the minimum acceptable potency level. Many factors affects the stability of a pharmaceutical product, including the stability of the active ingredient(s); the potential interaction between active constituents; the manufacturing process; the dosage form; the container/closure system; the environmental conditions encountered during shipment, storage and handling; and the period of time between its manufacture and usage. Environmental properties such as heat, light, and moisture, as well as chemical factors like oxidation, reduction, hydrolysis or racemisation, also play vital role in pharmaceutical stability. Stability indicating methodology is validated quantitative analytical method that can detect changes with time in chemical, physical or microbiological properties of drug substances and drug product and that are specific so that contents of active ingredients, degradation products and other components of interest can be accurately analyzed without interference.

3 Ampicillin:

NH2 N S

O N O OH

O Ampicillin sodium is derived from the penicillin nucleus, 6-aminopenicillanic acid. Chemically, it is monosodium (2S, 5R, 6R)-6-[(R)-2-amino-2-phenylacetamido]-3, 3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2-carboxylate and is a beta-lactam antibiotic that has been used extensively to treat bacterial infections since 1961. It is considered part of the amino penicillin family and is roughly equivalent to amoxicillin in terms of spectrum and level of activity. It can sometimes result in allergic reactions that range in severity from a rash (e.g. patients with mononucleosis) to potentially lethal anaphylaxis.

Sulbactum: O O S

H Sulbactam sodium is a derivative of the basic penicillin nucleus. Chemically, sulbactam sodium is sodium penicillinate sulfone; sodium (2S, 5R)-3,3-dimethyl-7-oxo-4-thia azabicyclo [3.2.0] heptane-2-carboxylate 4,4-dioxide. Sulbactam is a molecule which is given in combination with beta-lactam antibiotics to inhibit beta-lactamase, an enzyme produced by bacteria that destroys the antibiotics. Sulbactam is an irreversible inhibitor of beta-lactamase; it binds the enzyme and does not allow it to interact with the antibiotic. Sulbactam is able to inhibit the most common forms of beta-lactamase but is not able to interact with the ampC cephalosporinase. Thus, it confers little protection against bacteria such as Pseudomonas aeruginosa, Citrobacter, Enterobacter, and Serratia, which often express this gene.

4 In this project work, attempt shall be made to carryout stability studies of ampicillin and sulbactum in parentral dosage form. Its stability studies are important to know its potency, as degradation products formed may be toxic and may deteriorate the patient’s health further.

6.2 Review of the Literature: . International conference on hormonisation1 gives guidelines for stability testing of new drug substances and products under stability guidelines. . Wildfeuer. A, Räder. K2 has reported Stability of beta-lactamase inhibitors and beta-lactam antibiotics in parenteral formulations as well as in body fluids and tissue homogenates. Comparison of sulbactam, clavulanic acid, ampicillin and amoxicillin. . Mushinsky.R. F et al3. have discussed Stability of sulbactam/ampicillin in diluents for parenteral administration. . Hoda Mahgoub and Fatma Ahmed Aly4 have developed UV-spectrophotometric determination of ampicillin sodium and sulbactam sodium in two-component mixtures. . Thomas D el al5. have reported Stability of cefepime and metronidazole prepared for simplified administration as a single product. . Lieve Okerman, Johan Van Hende and Lieven De Zutter6. Stability of frozen stock solutions of beta-lactam antibiotics, cephalosporins, tetracyclines and quinolones used in antibiotic residue screening and antibiotic susceptibility testing. . Vijay Kumar, Hemant Bhutani and Saranjit Singh7. Have discussed ICH guidance in practice: Validated stability-indicating HPLC method for simultaneous determination of ampicillin and cloxacillin in combination drug products. . Erdal Dinçand Dumitru Baleanu8 has developed a new fractional wavelet approach for the simultaneous determination of ampicillin sodium and sulbactam sodium in a binary mixture. . R. Chadha. N.Kashidand D.V.S. Jain9. have reported Micro calorimetric evaluation of the in vitro compatibility of amoxicillin/clavulanic acid and ampicillin/sulbactam with ciprofloxacin. . A. Wildfeuer and K. Räder10. Stability of β-lactamase inhibitors and β-lactam antibiotics in parenteral dosage forms and in body fluids and tissue homogenates: a comparative study of sulbactam, clavulanic acid ampicillin and amoxicillin.

5 . F. Fitouss et al11. have investigated Inhibitory activity of various concentrations of clavulanic acid, tazobactam and sulbactam against 117 beta lactamase producing enterobacteriaceae strains. . Sonja Riediker, Andreas Rytz and Richard H. Stadler12 have studied Cold-temperature stability of five β-lactam antibiotics in bovine milk and milk extracts prepared for liquid chromatography–electrospray ionization tandem mass spectrometry analysis.

. Eric Verdon et al13. Stability reported has of penicillin antibiotic residues in meat during storage Ampicillin. . Reza Oliyai and Siegfried Lindenbaum14 have studied Stability testing of pharmaceuticals by isothermal heat conduction calorimetry: Ampicillin in aqueous solution .

6.3 OBJECTIVES OF THE STUDY:

In the proposed work attempts shall be made to:

 To study the effect of temperature and relative humidity with time on the physical and chemical properties of -lactam antibiotics in finished product.

 To establish the shelf life for the finished product and to recommend the storage conditions.

7.  To perform stability studies of combination of  lactam antibiotics in a formulation.

MATERIALS AND METHODS:

7.1 SOURCE OF DATA:

 Literature survey was done in the Library of P.E.S.College of Pharmacy.  Reference from STRIDES ARCOLAB LIMITED.  The relevant data was obtained from Internet facilities at P.E.S.College of Pharmacy, Bangalore-50.  Department of Drug Testing Laboratory, Bangalore.

6  Indian Institute of Science, Bangalore.

7.2 METHOD OF COLLECTION OF DATA:

All stability studies experiments shall be carried out at ATRIDES ARCOLAB LIMITED, in Bilekahalli. They are adequately equipped with necessary analytical setup to carry out stability studies.

7.3 Does the study require any investigation to be conducted on patients or Animals? -: No: -

7.4 Has ethical clearance been obtained from your institution in case of above?

-: Not applicable: -

References:

1. ICH, Stability guidelines, Q1A (R2), stability testing of new drug substance and products; 2003: 1-20. 2. WildfeuerA, Räder K. Stability of beta-lactamase inhibitors and beta-lactam antibiotics in parenteral formulations as well as in body fluids and tissue homogenates. Comparison of sulbactam, clavulanic acid, ampicillin and amoxicillin, 1991; 41(1):70-3. PMID: 2049115 [PubMed - indexed for MEDLINE]. 3. Mushinsky RF, Reynolds ML, Nicholson CA, Crider LL, Forcier GA Stability of sulbactam/ampicillin in diluents for parenteral administration, 1986; 8:523-7. PMID: 3026000 [PubMed - indexed for MEDLINE].

4. Hoda Mahgoub and Fatma Ahmed Aly. UV-spectrophotometric determination of ampicillin sodium and sulbactam sodium in two-component mixtures, J :Ph. and Biomed.

7 Ana, 1998;17 (8): 1273-1278.

5. Thomas D. Nolin, Daniel A. Lambertand Robert C. Owens, Jr. Stability of cefepime and metronidazole prepared for simplified administration as a single product, 2006; 56(2): 179-184. [Pub Med-indexed for MEDILINE] 6. Lieve Okerman, Johan Van Hende and Lieven De Zutter. Stability of frozen stock solutions of beta-lactam antibiotics, cephalosporins, tetracyclines and quinolones used in antibiotic residue screening and antibiotic susceptibility testing, Analytica Chimica Acta,2006;586(1- 2):284-288. 7. Vijay Kumar, Hemant Bhutani and Saranjit Singh. ICH guidance in practice: Validated stability-indicating HPLC method for simultaneous determination of ampicillin and cloxacillin in combination drug products. J :Ph. and Biomed. Ana. 2006;43(2):769-773. 8. Erdal Dinç and Dumitru Baleanu. A new fractional wavelet approach for the simultaneous determination of ampicillin sodium and sulbactam sodium in a binary mixture, J: of Pharm. and Biomed Ana, 2006; 63(3):631-638. 9. R. Chadha, N. Kashid and D. V. S. Jain. Microcalorimetric evaluation of the in vitro compatibility of amoxicillin/clavulanic acid and ampicillin/sulbactam with ciprofloxacin. J: of Pharm and Biomed Ana, 2004; 36(2): 295-307. 10. A. Wildfeuer and K. Räder Stability of β-lactamase inhibitors and β-lactam antibiotics in parenteral dosage forms and in body fluids and tissue homogenates. J: Pharm and Biomed Ana, 1996; 6(1): S31-S34. 11. F. Fitoussi, G. Arlet, I. Casin, Ph. Lagrange and A. Philippon Inhibitory activity of various concentrations of clavulanic acid, tazobactam and sulbactam against 117 betalactamase producing enterobacteriaceae strains. J :Pharm and Biomed Ana, 1993; 23(8): 8-21. 12. Sonja Riediker, Andreas Rytz and Richard H. Stadler. Cold-temperature stability of five β- lactam antibiotics in bovine milk and milk extracts prepared for liquid chromatography– electrospray ionization tandem mass spectrometry analysis. J: of Pharm and Biomed Ana,

8 2004; 1054(1-2): 359-363.

13. Eric Verdon, Régine Fuselier, Dominique Hurtaud-Pessel, Pierrick Couëdor, Nathalie Cadieu and Michel Laurentie. Stability of penicillin antibiotic residues in meat during storage Ampicillin. J : of Chrom A, 2000; 882(1-2): 135-143. 14. Reza Oliyai and Siegfried Lindenbaum. Stability testing of pharmaceuticals by isothermal heat conduction calorimetry: Ampicillin in aqueous solution. International J: of Ph, 1991; 73(1): 33-36.

15. Websites:

www.sciencedirect.com. www.pubmed.com. www.medline.com. www.chemindustry.com. www.google.com.

9 9. SIGNATURE OF THE CANDIDATE

(Siddalinga Prasad. N. C)

10. REMARKS OF THE GUIDE

11. NAME AND DESIGNATION OF

11.1 GUIDE Mr. Venkatesh Prasad, Assistant Professor, Department of Pharmaceutical Analysis, P.E.S.College of Pharmacy, Hanumanthanagar, Bangalore-560050

11.2 SIGNATURE

11.3 CO-GUIDE NOT APPLICABLE 11.4 SIGNATURE

11.5 HEAD OF THE DEPARTMENT Dr. Nagaraj. G Professor & HOD. Department of Pharmaceutical Analysis, P.E.S.College of Pharmacy, Hanumanthanagar, Bangalore-560050

11.6 SIGNATURE

12. REMARKS OF THE PRINCIPAL

SIGNATURE OF THE PRINCIPAL Prof. Dr. S. Mohan Principal, P.E.S.College of Pharmacy, Hanumanthanagar, Bangalore-50.