Development and Validation of a HPLC Method for Simultaneous Determination of Dapivirine and DS003 in Combination Microbicide Tablet

Abstract

A reverse phase liquid chromatographic analytical method was developed for the simultaneous determination of Dapivirine and DS003 content in tablet dosage form. The chromatographic separation was achieved by using a C 18 column with mobile phase containing a gradient mixture of 0.05% v/v trifluoroacetic acid in water as mobile phase A and acetonitrile as mobile phase B at a flow rate of 0.5 ml/min. Both of the analytes were quantified with a UV detector at 245 nm. Dapivirine and DS003 were subjected to the stress conditions of oxidation, acid and base hydrolysis and thermolysis. The analytes were found to be stable under acidic and thermal conditions. Dapivirine was not significantly affected by base hydrolysis but was severely affected by oxidative conditions. DS003 was significantly affected by both oxidation and base hydrolysis. The stability-indicating capability of this method was demonstrated by adequate separation of the degradation peaks from those of the actives in the stress degraded samples. The method was validated for linearity, specificity, system suitability, precision and accuracy in accordance with ICH guidelines. The proposed method was applied to quantitate Dapivirine and DS003 in drug-excipient compatibility studies, assay of the tablets and stability studies of Dapivirine and DS003 tablets.

Validation of a stability-indicating spectrometric method for the determination of sulfacetamide sodium in pure form and ophthalmic preparations

“…In such type of studies, degradation up to 15%–20% is generally considered acceptable as higher degradation may produce different degradation products which are generally not encountered during routine stability studies or shelf-life of the product. [ 27 28 ] The forced degradation does not produce any measurable thermal or photochemical change to affect the accuracy of the assay method. However, on prolonged light exposure (at least for 200 h at 70°C–90°C), SN is known to be further photo-oxidized to azobenzene-4,4-disulfoxamide which exhibits absorption maxima at 320 nm.…”

Microbicide Dosage Forms

“…The International Partnership for Microbicides (IPM) developed a combination tablet containing the NNRTI dapivirine (TMC120) and a gp120 inhibitor DS003 (Gupta et al 2011). This formulation utilized standard solid dosage form excipients (Table 2) and direct blend methodology, followed by standard tableting techniques to produce an ovule-shaped tablet.…”

Development and validation of a HPLC method for the assay of dapivirine in cell-based and tissue permeability experiments

“…An initial method (Table S1; Supplementary Data) was established after performing different screening runs based on previously reported HPLC-UV methods for assaying dapivirine [15][16][17][18][19]. The column used in this work was selected specifically due to its low silanol activity [25] since dapivirine is known to interact strongly with silanol groups, resulting in peak tailing [18].…”

“…A few high-performance liquid chromatography (HPLC) methods with UV quantification have been briefly described in the literature for dapivirine mainly for evaluating drug content or drug release from different pharmaceutical dosage forms/delivery systems [15][16][17]. Two HPLC-UV methods validated according to ICH guidelines have been reported by our group for polymeric nanoparticles [18] and by Gupta et al for combination vaginal tablets [19]. Also, liquid chromatography tandem mass spectrometry (LC-MS/MS) methods have been used for clinical study samples (genital tissue, cervicovaginal lavage and plasma) [6,7,20] but no details were provided on the method procedure.…”