Development and Validation of Simple Analytical Methods for the Estimation of Zidovudine

Development and Validation of Simple Analytical Methods for the Estimation of Zidovudine

6.
7.
8. / ENCLOSURE-1
BRIEF RESUME OF THE INTENDED WORK:
6.1 GENERAL DISCUSSION :
Tenofovir disproxil fumarate [1]belongs to a class of antiretroviral drugs known as nucleotide analogue reverse transcriptase inhibitors (nRTIs), which block reverse transcriptase, an enzyme crucial to viral production in HIV-infected people. It inhibits the activity of HIV reverse transcriptase by competing with the natural substrate deoxyadenosine 5’-triphosphate and, after incorporation into DNA, by DNA chain termination.
STRUCTURAL FORMULA :

Nomenclature : 9-[(R)-2-[[bis[(iso propoxy carbonyl)oxy](methoxy]phospho phenyl]
Methoxy] propyladenine fuamarate.
Molecular formula : C19H30N5O10P • C4H4O4
Molecular weight : 635.52 g/mol
Characteristics : White to half white crystalline powder
Category : Antiretroviral
Solubility : Soluble in water
Functional group : Amine group
6.2NEED FOR THE STUDY :
Analytical method developmentshould beperformed to the extent that itis sufficient for its intendedpurpose and cost effective.Upon successful completion of method development, the proposed method will be validated to show the suitability for its intended use. Finally, the validated method will betransferred to the quality control laboratory in preparation for the launch of thedrug substance or drug product.
Whether it may be a drug substance or a drug product or the final product will needto be analyzed to assess its potency or strength. Extensive literature survey reveals that only few analytical methods have been reported for the estimation of Tenofovir disproxil fumarateand are UV-Derivative Spectrophotometry [2,3] and RP-HPLC [4, 5,6] methods. Apart from above no other spectroscopic methods such as UV/Vis spectroscopy, Difference spectrophotometric method, HPLC by using internal standard etc., were reported for this compound.
Hence there is a need for the development of newer, simple, sensitive, rapid, accurateand reproducible analytical methods for the routine estimation of Tenofovir disproxil fumaratein pure and pharmaceutical dosage form.
6.3 REVIEW OF LITERATURE :
  1. Mirna EL Barkil, et al [2]., studied on Relevance of a combined UV and single mass spectrometry detection for the determination of Tenofovir disproxil fumarate in human plasma by HPLC in therapeutic drug monitoring.
  2. Gnanaraju, et al [3]., studied on method for development of Tenofovir as API and tablet dosage forms by UV spectroscopy.
  3. Appalaraju N,[4].,Studied on Simultaneous Estimation of Tenofovir disproxil fumerate, Emtricitabine & Efavirenz in tablet dosage forms by Isocratic-RP-HPLC.
  4. Kandagal P B, et al [5]., Studied on Tenofovir disproxil fumarate in Pharmaceutical Formulations and Spiked Human Plasmaby reverse phase high performance liquid chromatography.
  1. Sentenac S, et al [6]., Studied on Sensitive determination of Tenofovir disproxil fumarate in human plasma samples using reversed-phase liquid chromatography.
  2. Naser L Rezk, et al [7]., Studied on Simultaneous quantification of Emtricitabine and Tenofovir disproxil fumarate in human plasma using high-performance liquid chromatography after solid phase extraction.
  3. Tracy king, et al [8]., Studied on Tenofovir disproxil fumarate diphosphate in human peripheral blood mononuclear cells by Liquid chromatography–tandem mass spectrometric method.
  4. Tom delahanti, et al [9]., Studied on Sensitive assay for determining plasma Tenofovir disproxil fumarate concentrations by LC/MS/MS.
6.4OBJECTIVES OF THE STUDY:
In view of the need for a suitable method for routine analysis ofTenofovir disproxil fumarate in formulations, attempts are being made to develop simple, precise and accurate analytical methods for estimation ofTenofovir disproxil fumarate and extend it for their determination in formulation.
In the proposed work attempts shall be made to:
  • To establish sensitive and accurate methods for the quantitative estimation of Tenofovir disproxil fumarate in pure and dosage form.
  • To validate the newly developed methods in accordance with the analytical parameters mentioned in the ICH guidelines.[11,12]
  • To apply the newly developed, validated analytical methods for quantitative estimation of Tenofovir disproxil fumarate in bulk and pharmaceutical dosage forms.
ENCLOSURE-II
MATERIALS AND METHODS:
MATERIALS:
  • All the chemical and reagents for the development of new analytical method to estimate Tenofovir disproxil fumarate will be of analytical grade.
  • The pure sample of Tenofovir disproxil fumarate for the research work will be produced from Shashan pharmaceuticals Pondicherry.
  • 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.
  • HPTLC instrument CamagLinomat 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 of Tenofovir disproxil fumarate in bulk and pharmaceutical dosage form.
  • Development of Analytical methods on Tenofovir disproxil fumarate shall 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 of Tenofovir disproxil fumarate the spectrophotometric methods like Derivative spectroscopy, Difference spectroscopy and Area under Curve method will be developed.
  • Chromatographic methods like RP-HPLC by using different flow rate And HPTLC by using economic solvent system will be developed.
  • Colorimetric methods will be developed depending upon the chemical nature (functional group, chromophore) of Tenofovir disproxil fumarate.
7.1.Source of Data:
  1. Library, Bharathi College of Pharmacy.
  2. E-library, Bharathi College of Pharmacy.
  3. Library IICT, Hyderabad.
  4. Library IISC, Bangalore.
  5. Library, RGUHS,Bangalore.
  6. Internet.
7.2.Method of Collection of Data:
DATA COLLLECTED FROM:
JOURNALS:
  1. Journal of Chromatography B.
  2. Journal of Pharmaceutical and Biomedical Analysis.
  3. E-Journal of Chemistry.
  4. Chromatographia.
  5. Chemical and Pharmaceutical Bulletin.
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?
NOTAPPLICABLE
ENCLOSURE-III
REFERENCES:
  1. URL:
  2. Mirna El Barkil, Marie-Claude Gagnieu, Jerome Guitton. Relevance of a combined UV and single mass spectrometry detection for the determination of Tenofovir disproxil fumarate in human plasma by HPLC in therapeutic drug monitoring. J chromatogr B2007;854(1-2):192-197.
  3. Gnanaraju G, Guptha AK, juyal V, Kumar P. Validated method for development of Tenofovir as API and tablet dosage forms by UV spectroscopy. J young pharm 2009; 1(4):351-355.
  4. Applaraju N. Simultaneous Estimation of Tenofovir disproxil fumarate disproxil Fumerate, Emtricitabine & Efavirenz in Tablet dosage forms by Isocratic-RP-HPLC. J pharm res 2009;2(6):97-103.
  5. Kandagla PB, Manjunatha DH, Seetharamappa J,Kalanura SS. RP- HPLC Method for the Determination of Tenofovirdisproxil fumarate in Pharmaceutical Formulations and Spiked Human Plasma. Anal Letrs 2008; 41(4):561-570.
  6. Sentenac S,Fernandez C, Thuillier A, Lechat P, Aymard G.Sensitive determination of Tenofovir disproxil fumarate in human plasma samples using RP-HPLC.J chromatogr B2003; 793(2):317-324.
  7. Naser L Rezk, Rustin D Crutchley, Angela D, Kashuba M.Simultaneous quantification of Emtricitabine and Tenofovir disproxil fumarate in human plasma using HPLC after solid phase extraction. J chromatogr B 2005; 822(1-2):201-208.
  8. Tracy King, Lane Bushman, Jennifer Kiser, Peter L Anderson, Michelle Ray, Thomas Delahunty. et al. LC/MS determination of Tenofovir disproxil fumarate diphosphate in human peripheral blood mononuclear cells. J chromatogr B 2006; 843(2):147-156.
  1. Tom Delahunty, Lane Bushman, Courtney V Fletcher. Sensitive assay for determining plasma Tenofovir disproxil fumarate concentrations by LC/MS/MS.J chromatogr B2006;830(1):6-12.
  2. Noel A Gomes, Vikas V vaidya, Ashuthosh Pudage. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for simultaneous determination of Tenofovirdisproxil fumarate and Emtricitabine in human plasma and its application to a bioequivalence studyJ Pharm Biomed Anal 2008;48(3):918-926.
  3. ICH, Q2A Text on validation of analytical procedures, Oct, 1991
  4. ICH, Q3BValidationofanalyticalprocedures:methodology,Nov,1996.

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