“DEVELOPMENT AND VALIDATION OF NEW ANALYTICAL METHODS FOR SIMULTANEOUS ESTIMATION OF DORZALAMIDE AND TIMOLOL IN BULK AND PHARMACEUTICAL DOSAGE FORM”

SYNOPSIS FOR

M. PHARM DISSERTATION

SUBMITTED TO

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

Ms.Akula Ramya

I - M. PHARM

DEPARTMENT OF PHARMACEUTICAL ANALYSIS

PESCOLLEGE OF PHARMACY

BENGALURU-560 050

(2010-2011)

UNDER THE GUIDENCE OF

Mr. VENKATESH PRASAD S.P.

Asst. Professor

DEPARTMENT OF PHARMACEUTICAL ANALYSIS

P.E.S.COLLEGE OF PHARMACY

BENGALURU-560 050

KARNATAKA

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BENGALURU.

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the candidate and address: / Ms. Akula Ramya
Department of Pharmaceutical Analysis,
P.E.S.College Of Pharmacy,
50 Feet Road, Hanumanthanagar,
BSK I- Stage,
Bengaluru-560 050
Permanent Address
Ms. Akula Ramya
D/O Akula Kiran kumar,
Ashok Nagar,
Guntur, Guntur Dt,
Andhra Pradesh-522002.
2. / Name of the institution: / P.E.S.College of Pharmacy
50 Feet Road, Hanumanthanagar,
BSK I-Stage,
Bengaluru-560 050
3. / Course of study and subject: / MASTER OF PHARMACY IN
PHARMACEUTICAL ANALYSIS
4. / Date of the admission: / 03/07/2011
5. / Title of the topic:
“DEVELOPMENT AND VALIDATION OF NEW ANALYTICAL METHODS FOR SIMULTANEOUS ESTIMATION OF DORZALAMIDE AND TIMOLOL IN BULK AND PHARMACEUTICAL DOSAGE FORM”
6. / BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY:
Glaucoma is an eye disorder in which the optic nerve suffers damage, permanently damaging vision in the affected eye and progressing to complete blindness if untreated. It is generally associated with increased intraocular pressure.
Intraocular pressure is mainly maintained by the liquid aqueous humor, which is produced by the ciliary body of the eye. Aqueous humor normally does not go into the posterior segment of the eye; it is kept out of this area by the lens and zonule of zinn. Instead it stays only in the anterior segment, which is divided into the anterior and posterior chambers. When the ciliary bodies produce the aqueous humor, it first flows into the posterior chamber bounded by the lens and the iris. It then flows through the pupil of the iris into the anterior chamber bounded by the iris and the cornea. From here it flows through a structure known as the trabecular meshwork to enter the normal body circulation.Ocular hypertension is the condition of elevated intraocular pressure. The two main mechanisms of ocular hypertension are an increased production of aqueous humor, or a decreased outflow of aqueous humor.Glaucoma can be roughly divided into two main categories, open angle and close angle glaucoma.
The combination ofdorzolamide and timolol are available in a single dosage form as ophthalmic eye drops. Dorzolamide is a topical carbonic anhydrase inhibitor and timolol is a beta-adrenergic receptor blocking agent. The combination have additive or synergistic role in the treatment of increased pressure in the eye caused by open angle glaucoma or a condition called hypertension of the eye when topical beta-blocker monotherapy is not sufficient.(1)
DRUG PROFILE:
Dorzalamide :

IUPAC name : (4S-trans)-4-(ethylamino)-5,6-dihydro-6-methyl-4H-thieno[2,3-b]thiopyran-2-sulfonamide7,7-dioxide.
Molecular Formula : C10H16N2O4S3
Molecular weight : 324.443g/mole
MECHANISM OF ACTION:
Carbonic anhydrase (CA) is an enzyme found in many tissues of the body including the eye. ItScatalyzes the reversible reaction involving the hydration of carbon dioxide and the dehydration of carbonicacid. In humans, carbonic anhydrase exists as a number of isoenzymes, the most active being carbonicanhydrase II (CA-II), found primarily in red blood cells (RBCs), but also in other tissues. Inhibition ofcarbonic anhydrase in the ciliary processes of the eye decreases aqueous humor secretion, presumabling by slowing the formation of bicarbonate ions with subsequent
reduction in sodium and fluid transport. Theresult is a reduction in intraocular pressure (IOP).
Elevated intraocular pressure is a major risk factor in the pathogenesis of optic nerve damage and
glaucomatous visual field loss. . Dorzolamide also accumulates in red blood cells as a result of CA-II binding, as CA-II is found predominantly in erythrocytes.
Timolol:

IUPAC name :(-)-1-(tert-butylamino)-3-[(4-morpholino-1,2,5-thiadiazol-3-yl)oxy]-2-propranol
Molecular formula: C13H24N4O3S
Molecular weight : 316.421g/mol
MECHANISM OF ACTION:
Timolol is a beta-1 and beta-2 (non-selective)adrenergic receptor blocking agent. It works by blocking beta-receptors in the eye. The pressure within the eyeball is naturally maintained by a continuous flow of liquid called aqueous humor through the eyeball. Timolol blocks beta-receptors that are found on the ciliary body and in the blood vessel. This causes the blood vessels to constrict, and reduces the amount of watery fluid that filters out of the blood vessels to form aqueous humor.
Extensive literature survey reveals that few analytical methods have been reported for the estimation of dorzolamideand timolol individually and in combination with other drug which includes spectrophotometry,derivative spectroscopy(2,3),TLC densitometry(4,5),HPTLC(6),HPLC(7,10).The literature review reveals only fewanalytical method for the simultaneous estimation of dorzolamideand timolol by first derivative UV spectrophotometry, ratio derivative spectrophotometry, TLC densitometry, RP-HPLC.
In view of the need for a suitable method for routine analysis in combined formulations, attempts are being made to develop simple, precise and accurate analytical methods for simultaneous estimation of title ingredients and extend it for their determination in formulation.Analytical validation is the corner stone of process validation. Without a proven measurement system it is impossible to confirm whether the manufacturing process has done what it purport to do. Hence, there is a need to validate the new methods developed.
6.2 REVIEW OF THE LITERATURE:
  • Sharath HM, Babu Jose G, Channabasavaraj KP, Modiya JS.have developed thedevelopment and validation of spectrophotometric methods for estimation of dorzolamide hydrochloride in bulk and pharmaceutical dosage forms.
  • Hafsa Mohammedi, Somashekar Shyale, and Shanta Kumar SM. have developed the physico-chemical characterization, UV Spectrophotometric method development and validation studies of timolol maleate.
  • Pritam SJ, Rahul NK, Hardik NJ and Sanjay JS.have developed the development and validation of TLC-densitometry method for simultaneous estimation of brimonide tartarate and timolol maleate in bulk and pharmaceutical dosage form.
  • Lories IB.have developed the application of TLC-densitometry, first-derivativeUV-spectrophotometry and ratioderivativespectrophotometry for the determination of dorzolamidehydrochloride and timolol maleate.
  • Sanjivani P, Kulkarni, Poornima D, Amin have developed the stability indicating HPTLC determination of timolol maleate as bulk drug and in pharmaceutical preparations.
  • Serap saglik. have developed the determination of dorzolamide in ophthalmic solutions using solid-phase extraction by high performance liquid chromatography.
  • Sharath HM, Channabasavaraj K, Jose GB, Jagadish SM.have developed stability indicating RP-HPLC method for analysis of dorzolamide hydrochloride in the bulk drug and its pharmaceutical dosage form.
  • Arun P, Murugesan SK, Nanjaian M.have developed the simultaneous estimation of bromonidine tartarate and timolol maleate in nanoparticles formulation by RP-HPLC.
  • Nagori BP, Amit Maru, Pankaj Muysuni and Subhash Guptahave developed the method development and its validation for simultaneous estimation of timolol maleate and dorzolamide hydrochloride in as API and in ophthalmic solution dosage form by RP-HPLC.
  • Marini RD, MatthijsN,Vander Heyden Y,Smeyers-Verbeke, Dehouck, Hoogmartens, Silvestre P,Ceccato A, Goedert PH,Saevels J, Herbots C ,Caliaro G, Herr aez-Hern R , Verd´u-Andr`es J, Camp´ıns-falc´o P, Van de Wauw W, De Beer, Boulanger B, Chiap P, Crommen J, Hubert PH have developed the collaborative study of an liquid chromatographic method for the determination of R-timolol and other related substances in S-timolol maleate.
6.3 OBJECTIVES OF THE STUDY:
In the proposed work attempts shall be made to:
  • Develop new analytical method for simultaneous estimation of dorzolamide and timolol in combination preferably by HPLC, HPTLC, and Spectroscopy.
  • Validate the proposed method in accordance with BP, USP and ICH guidelines for the intended analytical application.
  • Apply the proposed methods for the analysis of drugs in their bulk and pharmaceutical dosage forms.
7.MATERIALS AND METHODS:
7.1SOURCE OF DATA:
  • Literature survey was done at P.E.S. College of Pharmacy using internet facilities
(RGUHS HELINET) and at Library.
  • Reference from library at RGUHS, Bengaluru.
  • Indian Institute of Sciences, Bengaluru.
7.2 METHOD OF COLLECTION OF DATA:
Analytical method development on Opthalmics drugs shall be carried out in collaboration with M/s STRIDES ARCOLAB LIMITED (Analytical service department). The analytical methods will be developed by using instruments like UV visible spectrometer, HPLC. The data so obtained is treated statistically to determine the compliance of the experimental result as per ICH and Pharmacopeial guidelines and for the routine use of the developed analytical methods in industry.
JOURNALS:
  1. Indian Journal of Pharmaceutical Sciences and Research
  2. Journal of Chromatography.
  3. Asian Journal of Chemistry.
  4. Analytical Chimica Acta.
  5. Indian Drugs.
  6. Journal of Pharmaceutical and Biomedical Analysis.
  7. International Journal of Pharmacy and Pharmaceutical Sciences
  8. International Journal of Recent Advances in Pharmaceutical Research
7.3DOES THE STUDY REQUIRE ANY INVESTIGATION TO BE CONDUCTED ON PATIENTS OR ANIMALS?
-: NO: -
7.4HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION
IN CASE OF ABOVE?
-: NOT APPLICABLE:
8.REFERENCES:
1. Scientific Diccusson.
2.Sharath HM, Babu JG, Channabasavaraj, Modiya JS. Development and Validation
of Spectrophotometric Methods for Estimation of Dorzolamide HCl in Bulk and Pharmaceutical Dosage Forms.IJPSR. 2007;2(4):948-53.
3. Mohammedi H, Shyale S. Physico-Chemial Charactesation, UV Spectrohotometri Method Development and Validation Studies Of Timolol Maleate. International Journal of Pharmaceutical Sciences Review and Research. [Research]. 2011;6(2):163-6.
4. Jain PS, Khatal RN, Jivani HN, Surana SJ. Development and Validation of TLC-densitometry Method for Simultaneous Estimation of Brimonidine tartrate and Timolol maleate in Bulk and Pharmaceutical Dosage Form. J Chromatograph Separat Techniq. 2011;2(3):1-5.
5. Bebawy, LI.Application of TLC-densitometry, first-derivative UV-spectrophotometry and ratio derivative spectrophotometry for the determination of dorzolamide hydrochloride and timolol maleate. Journal of Pharmaceutical and Biomedical Analysis. 2002:737-46.
6.Sanjivani P, Kulkarni, Poornima D, Amin. Stability indicating HPTLC determination of timolol maleate as bulk drug and in pharmaceutical preparations. Journal of Pharmaceutical and Biomedical Analysis. 2000;23:983-7.
7.Serap S. Determination of dorzolamide in ophthalmic solutions using solid-phase extraction by high-performance liquid chromatography. Acta pharmaceutica turcica. 2000;62(2,3):98-100.
8.Sharath HM, Channabasavaraj K, Jose G, Babu, Jagadish S, Modiya. Stability Indicating RPHPLC Method for Analysis of Dorzolamide HCl in the Bulk Drug and its Pharamceutical Dosage Form. Int J Pharm pharm sci. 2011;3(3):100-5.
9.Phogat A, Kumar MS, Mahadevan N. Simultaneous Estimation of Brimonidine Tartrate and Timolol Maleate in Nanoparticles Formulation by RP-HPLC. International Journal of Recent Advances in Pharmaceutical Research. 2011:31-6.
10.Nagoria BP, Muysunic P, Guptad S. Method Development and Its Validation for Simultaneous Estimation of Timolol Maleate and Dorzolamide Hydrochloride in as API and In Ophthalmic Solution Dosage Form by RPHPLC. Journal of Chemical and Pharmaceutical Research. 2011:866-74.
11. Marini RD, Matthijs N, Vander HY,Smeyers VJ, Dehouck P, Hoogmartens J, et al. Collaborative study of an liquid chromatographic method for the determination of R-timolol and other related substances in S-timolol maleate. Analytica Chimica Acta. 2005;546:182-92.
Websites:







9. / Signature of the candidate / ( Akula Ramya)
10. / Remarks of the guide
11. / Name and Designation of
11.1 Guide / Mr. Venkatesh Prasad S.P
Assistant Professor
Department of Pharmaceutical Analysis,
P.E.S.College of Pharmacy,
Hanumanthanagar, Bengaluru-560 050
11.2 Signature
11.3 Head of the department / Dr. Nagaraj
Professor & Head
Department of Pharmaceutical Analysis
P.E.SCollege of Pharmacy, Hanumanthanagar,
Bengaluru-560 050
11.4 Signature
Remarks of principal
12. / Name and designation Chairman and Principal
(Forwarded for approval) / Prof. Dr. S. Mohan
Principal & Director,
P.E.SCollege of Pharmacy, Hanumanthanagar,
Bengaluru-560 050
12.1 Signature

1