The DEVELOPMENT OF A VALIDATED STABILITY INDICATING LC-MS METHOD FOR THE DETERMINATION OF TENOFOVIR DISOPROXIL FUMARATE USING QUALITY BY DESIGN APPROACH

Abstract

Objective: To develop a validated Liquid Chromatography-Mass Spectrometry (LC-MS) method through Quality by Design (QbD) approach for the determination of tenofovir disoproxil fumarate (tenofovir DF) in bulk, pharmaceutical formulation and stress conditions.
 Methods: The analytical target profile (ATP) was to develop a simple, accurate, precise, specific, and robust method with the Critical Quality Attribute (CQA) being retention time, theoretical plate and peak tailing. Derived from the risk assessment studies, four high-risk factors were screened using resolution V irregular fraction design. Organic modifier strength, flow rate and injection volume were further optimized using Box-Behnken design (BBD). The optimized method condition was validated and applied for the determination of the analyte in pharmaceutical formulation. The stability of the analyte in stress conditions was determined. All experiments were performed using reverse-phase Acquity UPLC BEH C18 (1.7 µm, 1 mm X 50 mm) column coupled in-line with Synapt G2 mass spectrometer in positive electrospray ionization (ESI) mode with electron multiplier analog-to-digital detector.
 Results: Optimized method condition eluted analyte at 3.40 min and the m/z 520 ratified the analyte peak. The method was validated and was found to be specific, linear in the range of 20-100 µg/ml (R2 = 0.9998) with accuracy between 98.71% - 101.17% and precise. The limit of detection and quantification were 5.50 µg/ml and 16.68 µg/ml respectively. Analyte degraded completely in acid and basic environment, however, stable in oxidative and photolytic conditions.
 Conclusion: The validated method developed through the systematic and rational QbD approach bespoke superior quality. The QbD approach not only justified the entire process but also eliminated uncertainties.

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 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.…”

The Optimization of Rp-HPLC Condition Using Response Surface Methodology Box-Behnken Design for Simultaneous Determination of Metformin HCL and Glimepiride in Spiked Plasma

“…Box-Behnken Design is a multivariate analysis technique that can minimize the time and costs for the optimization process [22][23][24]. It was applied to various studies, like the isolation process, development of drug formulation, and optimization of chromatographic conditions [25][26][27]. Optimization of HPLC conditions using BBD has been applied for the analysis of various samples [28][29][30].…”