Development and Validation of Rp-Chiral HPLC Method for Quantification of (S)-Isomer in Tenofovir Disoproxil Fumarate

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 the 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.

Rp-HPLC (Stability-Indicating) Based Assay Method for the Simultaneous Estimation of Doravirine, Tenofovir Disoproxil Fumarate and Lamivudine

“…The flow rate remains constant all along the trails at 1.0 ml per min. During trails, the results were checked for values of tailing factor, peak areas, theoretical plates and resolution for the The method presented was selective for the combined assay of LMV, TFF and DRV in tablets without interruption with diluent solvent system components and tablet excipients [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The coefficients of regression higher than 0.99 indicating a lined correlation between the LMV, TFF, and DRV concentration and their respective area responses [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30].…”

“…During trails, the results were checked for values of tailing factor, peak areas, theoretical plates and resolution for the The method presented was selective for the combined assay of LMV, TFF and DRV in tablets without interruption with diluent solvent system components and tablet excipients [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The coefficients of regression higher than 0.99 indicating a lined correlation between the LMV, TFF, and DRV concentration and their respective area responses [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The method was sufficiently sensitive for the combined assay of LMV, TFF and DRV in tablets because of low values of quantification and detection limits [14][15][16][17][18][19][20][21][22][23][24][25][26][27...…”

“…The coefficients of regression higher than 0.99 indicating a lined correlation between the LMV, TFF, and DRV concentration and their respective area responses [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The method was sufficiently sensitive for the combined assay of LMV, TFF and DRV in tablets because of low values of quantification and detection limits [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The precision results were below 2% RSD.…”

“…The precision results were below 2% RSD. The reports indicated that the assay method was precise for the LMV, TFF and DRV combined analysis [14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30]. The robustness results were below 2% RSD.…”

“…To quantify LMV, spectrophotometry [14,15] and HPLC [16][17][18][19] based methods were proposed. TFF was quantified using spectrophotometry [20], HPLC [21][22][23][24] and LC-MS [25,26] based methods. To determine DRV, LC-MS [27] based method was proposed.…”