Development and validation of an HPLC‐UV method for quantification of elvitegravir and two other new antiretrovirals, dolutegravir and rilpivirine, in the plasma of HIV‐positive patients

Abstract

Therapeutic drug monitoring may be crucial in selected clinical conditions for the management of HIV infection. In recent years, new antiretrovirals have been introduced and in particular elvitegravir (EVG) is now recommended for first-line and simplification treatment as well as dolutegravir (DTG) and rilpivirine (RPV). The aim of this study was to develop and validate a high-performance liquid chromatography-ultraviolet (HPLC-UV) method for determining EVG and new antiretrovirals DTG and RPV in human plasma. Solid-phase extraction was applied to a 600 μL plasma sample. Chromatographic separation of the three drugs and internal standard was achieved with a gradient of acetonitrile and phosphate buffer on a C reverse-phase analytical column with a 20 min analytical run time. EVG and DTG were detected at 265 nm and RPV at 290 nm. Mean intra- and inter-day precisions were < 10%; the mean accuracy was <15%. Extraction recovery ranged between 105 and 82% for the drugs analyzed. Calibration curves were optimized according to the expected ranges of drug concentrations in patients; the coefficient of determination was >0.997 for all drugs. This method allows for monitoring EVG, DTG and RPV in the plasma of HIV-positive patients using HPLC-UV.

Development and validation of an HPLC method for quantification of dolutegravir in human plasma

“…However, due to the high cost of equipment, these methods are difficult to be adopted in resource‐limited areas. Less costly HPLC–UV methods (Cozzi et al, 2016; Tempestilli et al, 2018) have been developed; however, they used solid‐phase extraction for the pre‐treatment of plasma samples and gradient elution program for chromatographic separation, which is both tedious and time‐consuming. The current study has established an HPLC–UV method for quantifying the concentration of DTG in human plasma, with less demanding HPLC techniques (i.e.…”

“…However, due to the high cost of equipment, these methods are difficult to be adopted in resource-limited areas. Less costly HPLC-UV methods (Cozzi et al, 2016;Tempestilli et al, 2018) Regarding the sample pre-treatment, we tried precipitation with acetonitrile or 5% perchloric acid, but there was no way to avoid the interference of blank plasma. We tried to extract DTG from plasma with chloroform, but its density is greater than that of water, thus making it inconvenient to transfer.…”

“…Several HPLC–MS/MS methods are now available to determinate DTG concentrations in plasma (Bennetto‐Hood et al, 2014; Bollen et al, 2019; Penchala et al, 2016; Simiele et al, 2017; Zheng et al, 2020), while the high cost of the equipment limits its application in less developed countries and regions. Cozzi et al (2016), Tempestilli et al (2018), and Wang et al (2016) developed chromatography coupled with ultraviolet detection methods for DTG determination, but they all used solid‐phase extraction for pre‐treatment of plasma, which is relatively costly compared with liquid–liquid extraction, and gradient elution process was applied for chromatographic separation, for which a double pump system is needed. Overall, there is a lack of method for quantification of DTG in human plasma that not only meets the basic requirements for quantification such as good sensitivity, accuracy, precision, but also has low demand for equipment and low reagent cost, thereby allowing it to be adopted in different levels of laboratories, especially poorly equipped ones.…”

“…Several HPLC-MS/MS methods are now available to determinate DTG concentrations in plasma (Bennetto-Hood et al, 2014;Bollen et al, 2019;Penchala et al, 2016;Simiele et al, 2017;Zheng et al, 2020), while the high cost of the equipment limits its application in less developed countries and regions. Cozzi et al (2016), Tempestilli et al (2018), and Wang et al (2016) developed chromatography coupled with ultraviolet detection methods for DTG determination, but they all used solid-phase extraction for pre-treatment of plasma, which is relatively costly compared with liquid-liquid extraction, and gradient elution process was applied for chromatographic separation, for which a double pump system is needed.…”

Simultaneous quantification of (E) and (Z) isomers of rilpivirine and four degradation products in bulk and tablets by reversed‐phase ultra‐high‐performance liquid chromatography and confirmation of all by molecular weight

“…A thorough literature review study demonstrated that different analytical techniques were used for the estimation of rilpivirine as a single content using spectrophotometric [4,5], HPTLC [6], HPLC [7][8][9][10], UPLC [11], LC-MS [12], and LC-MS/MS [13][14][15] in different matrixes (bulk, tablets, nanoparticles, saliva, serum, and human plasma). The literature also revealed the number of methods including RP-HPLC [16][17][18][19][20][21][22][23][24][25][26], HPLC-MS/MS [27][28][29][30][31], RP-UPLC [32][33], and UPLC-MS/MS [34][35][36] for the determination of rilpivirine in the presence of other antiretroviral drug molecules. To the best of our knowledge, there are only three reported works for the determination of rilpivirine with limited impurities and or in presence of degradation products in pharmaceutical samples by HPLC.…”