Development and Validation of HPTLC Method for Simultaneous Estimation of Emtricitabine, Rilpivirine and Tenofovir Disoproxil Fumarate in Combined Dosage form

Abstract

This study describes the development and validation of high performance thin layer chromatographic (HPTLC) method for the simultaneous estimation of Emtricitabine (EMT), Rilpivirine (RPV) and Tenofovir disoproxil fumarate (TFV) in combined dosage form. Chromatographic separation of these drugs was performed on aluminum plates precoated with silica gel 60 F254 as the stationary phase using solvent system consisted of chloroform: ethyl acetate: methanol: glacial acetic acid (5:2:1:0.1 v/v/v/v). The densitometric analysis was carried out in absorbance mode at 272 nm. The drugs were satisfactorily resolved with Rf values of 0.28 ± 0.02, 0.70 ± 0.02 and 0.52 ± 0.04, respectively. The method was validated according to the International Conference of Harmonization (ICH) guidelines. The calibration curves were linear over the (r 2 > 0.999) concentrations range from 600-2400 ng band -1 for Emtricitabine, 50-300 ng bands -1 for Rilpivirine and 600-3600 ng band -1 for Tenofovir disoproxil fumarate. The method showed accuracy of 100.01%, 100.32% and 100.14% and percentage assay of 99.91%, 98.72% and 99.34% for Emtricitabine, Rilpivirine and Tenofovir disoproxil fumarate, respectively. Percentage relative standard deviation (<2%) was found for both precision and robustness study showing that the proposed method was precise, specificity, robust and stable in accordance with ICH guidelines.

Quantitative Analysis of Emtricitabine in Dosage Forms Using Green RP-HPTLC and Routine NP-HPTLC Methods—A Contrast of Validation Parameters

“… 33 On the other hand, the precision and accuracy of all the reported HPTLC methods were found to be much better than the routine NP-HPTLC method. 29 − 34 The accuracy of literature HPTLC methods was found to be similar to the green RP-HPTLC method. Although, the precision of the literature HPTLC methods was found to be superior to the green RP-HPTLC method, but the precision values for the literature and green RP-HPTLC method were within the limit of ICH guidance (less than 2%).…”

“…However, the lower limit of quantification (LLOQ) of these methods was much lower than the green and routine densitometry method in this study. [29][30][31]34 The linearity for the other two literature HPTLC methods has been reported as 30−1100 ng/band 32 and 40− 400 ng/band, 33 respectively. The accuracy and precision of these two methods were also within the limit of ICH guidance.…”

“…Three different validation parameters including linearity, accuracy, and precision of the green and routine densitometry methods were compared with literature values of densitometry methods. The linearity for four of the literature HPTLC methods has been reported as 200–1000 ng/band, 29 200–2200 ng/mL, 30 80–560 ng/band, 31 and 400–2400 ng/band, 34 respectively. The accuracy and precision of these methods were within the limit of ICH guidance.…”

“… 27 , 28 Several high-performance thin-layer chromatography (HPTLC) techniques have also been used for the quantitative analysis of ECT in its single and combined dosage forms. 29 − 34 However, there is no single study and report on green reversed-phase HPTLC (RP-HPTLC) densitometry analysis of ECT in pharmaceutical dosage forms and biological samples. The reported HPTLC methods for the quantitative analysis of ECT have not considered the environmental impact.…”

“…The reported HPTLC methods for the quantitative analysis of ECT have not considered the environmental impact. 29 − 34 Recently, researchers are focusing much attention on green HPTLC methods for the quantitative analysis of drugs/pharmaceuticals in dosage forms and biological samples. 35 − 38 In addition, green HPTLC methods present many advantages compared to routine HPTLC methods of analysis.…”