6.
7.
8. / ENCLOSURE-I
BRIEF RESUME OF THE INTENDED WORK:
6.1 GENERAL DISCUSSION :
Rufinamide [1,2] is an anticonvulsant. It is used in combination with other medication and therapy to treat Lennox–Gastaut syndrome and various other seizure disorders. It is a triazole derivative, it is presumed to involve stabilization of the sodium channel inactive state, effectively keeping these ion channels closed. Although the direct mechanism of action may be different, several other antiepileptic agents also stabilize a sodium channel inactive state. It is also being investigated as a treatment for anxiety disorders.
Molecular Structure :

Nomenclature : 1-(2,6-difluorobenzyl)-1H-1,2,3-triazole-4-carboxamide.
Molecular formula : C10H8F2N4O
Molecular weight : 238.19g / mol.
Characteristics : White to off-white crystalline powder.
Solubility : Practically insoluble in water, slight solubility in tetra hydro furan
(THF) andmethanol, very slight solubility in ethanol and
acetonitrile, soluble in dimethyl sulfoxide (DMSO).
Category : Anticonvulsant drug.
Functional groups : Amide group, secondary and tertiary amino groups.
6.2 NEED FOR THE STUDY :
Modern analytical chemistry generally requires precise analytical measurements at very low concentrations, with a variety of instruments. Analytical methods are required to characterize drug substances and drug products composition during all phases of pharmaceutical development. Control of the chemical environment is often necessary to ensure that the analyte is measured in the desired form & to minimize the effect of interference. Development of methods to achieve the final goal of ensuring the quality of drug substances and drug products must be implemented in conjunction with an understanding of the chemical behavior and physicochemical properties of the drug substance. This determination requires highly sophisticated instruments and methods like Spectrophotometer, HPLC and HPTLC etc.
Extensive literature survey reveals that no analytical methods that have been reported for the estimation of Rufinamidein pure and dosage form which includes Spectrophotometry,HPLC andHPTLC.
Hence there is a need for the development of newer, simple, sensitive, rapid, accurate and reproducible analytical methods for the routine estimation of Rufinamidein bulk and pharmaceutical dosage form.
6.3REVIEW OF LITERATURE :
  1. Manuela Contin, et al[3]., have developedan implementation methodfor newer antiepileptic drugs (AEDs) in plasma of patients with epilepsy using high performance liquid chromatography (HPLC) with UV detection.HPLC analysis was carried out on a Synergi 4 μm Hydro-RP, 250 mm × 4.6 mm I.D. column. The mobile phase was a mixture of potassium dihydrogen phosphate buffer (50 mM, pH 4.5), acetonitrile and methanol (65:26.2:8.8, v/v/v) at an isocratic flow rate of 0.8 mL/min. The UV detector was set at 210 nm. The chromatographic run lasted 19 min. Commonly coprescribed AEDs did not interfere with the assay. The present study describes the simple and validated method for Rufinamide determination in plasma of patients with epilepsy.
  1. M. C. Rouanet al[4].,have developed a practice of solid-phase extraction and protein precipitation in the 96-well format combined with high-performance liquid chromatography–ultraviolet detection for the analysis of drugs in plasma and brain.C18 Empore 96-well extraction disc plates have been employed for the analysis of three drugs with different polarities in plasma in conjunction with HPLC–UV, rufinamide, ICL670 and an anticonvulsant agent (AA1) in an early stage of development. The methods were used routinely, one plate per analysis day being processed, resulting in increase in sample throughput and saving in solvents.
6.4 OBJECTIVES OF THE STUDY:
The study ofRufinamidemakes an attempt to establish sensitive and accurate methods for the estimation ofRufinamidein pure and tablet dosage form.
In view of the need for a suitable method for routine analysis ofRufinamidein formulations, attempts are being made to develop simple, precise and accurate analytical methods for estimation ofRufinamideand extend it for their determination in formulation.
In the proposed work attempts shall be made to:
To develop new, simple, sensitive, accurate, and economical analytical methods for estimation ofRufinamidein bulk and pharmaceutical dosage forms preferably by HPLC, HPTLC and Spectroscopy.
To validate the newly developed methods in accordance with the analytical parameters mentioned in the ICH guidelines.[5],[6].
To apply the newly developed, validated analytical methods for quantitative determination ofRufinamidein bulk and pharmaceutical dosage forms
ENCLOSURE-II
MATERIALS AND METHODS:
MATERIALS:
All the chemical and reagents for the development of new analytical method to estimateRufinamidewill be of analytical grade.
The pure sample ofRufinamidefor the research work will be produced from Ranbaxy Pharma, Ahmedabad.
Spectral measurement will be carried out using Shimadzu UV-VIS Spectrophotometer 1800.
Shimadzu HPLC model containing LC-20AT (VP series) pump, variable wavelength programmable UV / VIS detector SPD-20A (VP series) will be used for HPLC method.
Camag Linomat V sample applicator and Camag TLC Scanner 3 equipped with Win-CAT software will be employed for the development and validation of new HPTLC method for the estimation ofRufinamidein bulk and pharmaceutical dosage form.
Development of Analytical methods onRufinamideshall be carried out at Department of pharmaceutical analysis, Bharathi College of Pharmacy. The college is equipped with necessary analytical set up to carry out desired work.
METHODS:
Based on solubility ofRufinamidethe UV spectrophotometric methods like Derivative spectroscopy [8], Difference spectroscopy and Area under Curve method will be developed and validated.
The RP-HPLC[9,10] methods will be developed for estimation of Rufinamide by using with lesser retention and runtime by using different columns and solvent systems.
The validated HPTLC[11] method will be made and the chromatographic conditions will be optimised.
The colorimetricmethods[12,13] will be developed and validated depending upon analytically important functional groups located in Rufinamide which is responsible for chemical derivatization.
For the selection and standardization of mobile phases to be used in HPLC/HPTLC different combination and ratio of solvent will be tried to obtain good optimum resolution.
Various data collected for the analytical method development either by spectrophotometry or by HPLC/HPTLC will be treated statistically to find its compliance with ICH or other official guidelines. Typical analytical parameters such as linearity, range, precision, specificity, accuracy, ruggedness, robustness and other system suitability parameters used in assay validation will be determined.
7.1.Source of Data:
  • Library, Bharathi College of Pharmacy.
  • E-library, Bharathi College of Pharmacy.
  • Library, RGUHS, Bangalore.
  • Library IISC, Bangalore.
  • Library IICT, Hyderabad.
7.2. Method of Collection of Data:
DATA COLLLECTED FROM:
JOURNALS:
  • Journal of Chromatography B.
  • Journal of Pharmaceutical and Biomedical Analysis.
  • International Journal of Pharmaceutical Analysis.
  • Journal of Chromatography B Biomedical Science Application.
  • Journal of epilepsia.
RELATED LINKS:





7.3. DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS?
NOT APPLICABLE.
7.4. HAS ETHICAL CLEARENCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
NOT APPLICABLE.
ENCLOSURE-III
REFERENCES:
  1. Manuela Contin, Susan Mohamed, Carmina Candela, Fiorenzo Albani, Roberto Riva,Agostino Baruzzi.Simultaneous HPLC–UV analysis of rufinamide, zonisamide, lamotrigine, oxcarbazepine monohydroxy derivative and felbamate in deproteinized plasma of patients with epilepsy.J Chromatogr B2010;878(3-4):461-465.
  2. M. C. Rouan, C. Buffet, L. Masson, F. Marfil, H. Humbert, G. Maurer.Practice of solid-phase extraction and protein precipitation in the 96-well format combined with high-performance liquid chromatography–ultraviolet detection for the analysis of drugs in plasma and brain. J Chromatogr B Biomed Sci Appl 2001; 754(1):45-55.
  3. ICH, Q2A Text on validation of analytical procedures, International conference on harmonization, Oct, 1994.
  4. ICH, Q3B Validation of analytical procedures: methodology, International conference on harmonization, Nov, 1996.
  5. ICH, Q2 (R1) Validation of Analytical Procedures: text and methodology; 2005.
  6. Gurudeep R. Chatwal, Sham K. Anand. Instrumental methods of chemical analysis. 5th ed. Hyderabad: HPH; 2002. p. 149-184.
  7. Yuri Kazakevich, Rosaria Lobrutto. HPLC for Pharmaceutical scientists. New York: CBS; 2007. P. 139-161.
  8. Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC method development. 2nd ed. New York: Wiley Inter Science; 1997. p. 1-50.
  9. Sethi PD. HPTLC: Quantitative analysis of pharmaceutical formulations. 1st ed. New Delhi: CBS; 1996. p. 53-57.
  10. Beckett AH, Stenlake JB. Pharmaceutical Chemistry. 4th ed. Part-two. New Delhi: CBS; 1997. p. 302-304.
  11. Hobart H. Willard, Lynne L. Meritt, John A. Dean, Frank A. Settle. Instrumental method of analysis. 7th ed. New Delhi: CBS; 1986. p. 159-177.