DEVELOPMENT AND VALIDATION OF RP-CHIRAL HPLC METHOD FOR QUANTIFICATION OF (S)-ISOMER IN TENOFOVIR DISOPROXIL FUMARATE


Suresh Babu Bodempudi, Ravi Chandra Babu Rupakula, Konda S. Reddy

Abstract


Objective: The main objective of present study was to develop and validate a reverse phase enantioselective chiral high performance liquid chromatographic method was developed for enantiomeric resolution of Tenofovir disoproxil fumarate; it decreases the HIV infection in human body. The method is specific, rapid, precise and accurate for the separation and determination of (S)-isomer in tenofovir disoproxil fumarate drug substance form.

Methods: The S-Isomer of Tenofovir disoproxil fumarate was resolved on a Chiral AGP (150 × 4.0 mm, 5 µm) column (L-41) using a mobile phase system containing 0.1 M ammonium acetate in water pH 6.8 with ammonia solution and methanol in the ratio of (85:15 v/v). The mobile phase was set at a flow rate of 0.8 ml/min and the volume injected was 10μl for every injection. The detection wavelength was set at 260 nm and the column temperature was set at 15 °C.

Results: The proposed method was productively applied for the quantitative determination of (S)-isomer in Tenofovir disoproxil fumarate drug substance form. The linear regression analysis data for calibration plots showed a good linear relationship over a concentration range of 0.125 to 3.75 µg/ml for (S)-isomer, 0.125-3.75 µg/ml for Tenofovir disoproxil fumarate. The mean values of the correlation coefficient were 0.999 and 0.999 for (S)-isomer and Tenofovir disoproxil fumarate. The method was validated as per the ICH guidelines. The detection limit (LOD) was about 0.05 µg/ml and quantitation limit (LOQ) was about 0.125 µg/ml for (S)-isomer and Tenofovir disoproxil fumarate. The relative standard deviation was found to be 0.78 % for (S)-isomer in Tenofovir disoproxil fumarate.

Conclusion: The developed and validated HPLC method and the statistical analysis showed that the method is repeatable and selective for the estimation of the (S)-isomer of the Tenofovir disoproxil fumarate drug substance.

Keywords


Tenofovir disoproxil fumarate, (S)-Isomer; RP-Chiral HPLC, Identification, Quantification and Validation

| PDF | HTML |

References


Neil MJO. Ed. by the Merck Index-An Encyclopedia of Chemicals, Drugs and Biological; Merck and Co. Inc; 2006.

Sweetman SC. Ed. by Martindale, the Complete Drug Reference; Pharmaceutical Press; 2007.

Keamey BP, Flherty JF, Shah J. Tenofovir disoproxil fumarate: clinical pharmacology and pharmacokinetics. Clin Pharmacokinet 2004;43:595-612.

Seshachalam U, Rajababu B, Haribabu B, Chandrasekhar KB. Enantiomeric separation of tenofovir on an achiral C18 column by HPLC using L-phenylalanine as a chiral mobile phase additive. J Liq Chromatogr Relat Technol 2008;31:410-20.

Kandagal PB, Manjunatha DH, Seetharamappa J, Kalanur SS. RP-HPLC method for the determination of tenofovir in pharmaceutical formulations and spiked human plasma. Anal Lett 2008;41:561-70.

Sentenac S, Fernandez C, Thuillier A, Lechat P, Aymard G. Sensitive determination of TNF in human plasma samples using reversed-phase liquid chromatography. J Chromatography B 2003;793:317-24.

Jullien V, Treluyer JM, Pons G, Rey E. Determination of TNF in human plasma by high-performance liquid chromatography with spectrofluorimetric detection. J Chromatography B 2003;785:377-81.

Sparidens RW, Crommentuyon KM, Scheelens JH, Beijnen JH. Liquid chromatographic assay for the antiviral nucleotide analogue TNF in plasma using derivatization with chloroacetaldehyde. J Chromatography 2003;791:227-33.

Delahunty T, Bushman L, Fletcher CV. Sensitive assay for determining plasma TNF concentrations by LC/MS/MS. J Chromatogr B: Anal Technol Biomed Life Sci 2006;830:6-12.

Massaki T, Yuichi K, Naoya O, Atsushi H, Kazuhide B, Tsuguhiru K. Determination of plasma tenofovir concentrations using a conventional LC-MS method. Biol Pharm Bull 2007;30:1784-6.

Bezy V, Morin P, Couerbe P, Lepen G, Agrofoglio L. Simultaneous analysis of several antiretroviral nucleosides in rat-plasma by high-performance liquid chromatography with UV using acetic acid/hydroxylamine buffer Test of this new volatile medium-pH for HPLC–ESI-MS/MS. J Chromatogr B: Anal Technol Biomed Life Sci 2005;821:132-43.

Shirkhedkar AA, Bhirud CH, Surana SJ. Application of UV-spectrophotometric methods for estimation of tenofovir disoproxil fumarate in tablets. Pakistan J Pharm Sci 2009;22:27-9.

ICH. Q2 (R1) Validation of Analytical Procedures: Text and Methodology. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, ICH harmonized Tripartite Guideline; 2005.

Kanarek AD. Analytical methods: a statistical perspective on ICH Q2A and Q2B guidelines for validation of analytical methods. BioPharm Int 2005;19:28-33.

ICH guidance on method validation; validation of analytical procedures Text and Methodology Q2 (R1); 2005

ICH harmonized tripartite guideline impurities in new drug substances Q3 A (R2); 2006.




About this article

Title

DEVELOPMENT AND VALIDATION OF RP-CHIRAL HPLC METHOD FOR QUANTIFICATION OF (S)-ISOMER IN TENOFOVIR DISOPROXIL FUMARATE

Keywords

Tenofovir disoproxil fumarate, (S)-Isomer; RP-Chiral HPLC, Identification, Quantification and Validation

DOI

10.22159/ijcpr.2017v9i6.23425

Date

14-11-2017

Additional Links

Manuscript Submission

Journal

International Journal of Current Pharmaceutical Research
Vol 9, Issue 6 (Oct-Nov), 2017 Page: 31-36

Online ISSN

0975-7066

Statistics

11 Views | 3 Downloads

Authors & Affiliations

Suresh Babu Bodempudi
GITAM University, Visakhapatnam, India, and Dr. Konda’s Life Sciences, Hyderabad

Ravi Chandra Babu Rupakula
GITAM University, Visakhapatnam, India, and Dr. Konda’s Life Sciences, Hyderabad

Konda S. Reddy
GITAM University, Visakhapatnam, India, and Dr. Konda’s Life Sciences, Hyderabad


Refbacks

  • There are currently no refbacks.