A novel quality by design approach for development and validation of a green reversed‐phase HPLC method with fluorescence detection for the simultaneous determination of lesinurad, febuxostat, and diflunisal: Application to human plasma

Abstract

A novel and eco‐friendly reversed‐phase HPLC method with fluorescence detection was developed for simultaneous estimation of two co‐administered antigout drugs (lesinurad and febuxostat) with diflunisal as a nonsteroidal anti‐inflammatory drug. Unlike routine methodology, the developed method was optimized using analytical quality by design approach. A full factorial design was applied to optimize the effect of variable factors on chromatographic responses. The chromatographic separation was performed using isocratic elution on the Hypersil BDS C18 column at 40°C. The mobile phase consisted of acetonitrile:potassium phosphate buffer (30.0 mM; pH 5.5, 32.2:67.8% v/v) pumped at a flow rate of 1.0 mL/min and injection volume of 20.0 μL was employed. The proposed method was able to separate the ternary mixture in <10 min. The calibration curves of diflunisal, lesinurad, and febuxostat were linear over concentration ranges of 50.0–500.0, 50.0–700.0, and 20.0–700.0 ng/mL, respectively. Recovery percentages ranging from 98.1 to 101.3% with % relative standard deviation of <2% were obtained upon spiking to human plasma samples, indicating high bioanalytical applicability. Furthermore, the method was found to be excellent green when it was assessed according to Green Analytical Procedure Index and analytical Eco‐Scale guidelines.

Optimization and validation of liquid–liquid extraction with low-temperature purification (LLE-LTP) for determining fluopyram fungicide in water samples using HPLC-DAD

“…The optimization of LLE-LTP of fluopyram in water samples was carried out through two-level (2 3 ) full factorial design (FFD), wherein all influencing independent factors with level combinations are simultaneously investigated. 20 The three parameters evaluated (see Table 1) were chosen from previous studies. 9,21 It was observed in these studies that these parameters had a decisive influence on the increase in the recovery percentage.…”

Quantification and Determination of Stability of Tylvalosin in Pig Plasma by Ultra-High Liquid Chromatography with Ultraviolet Detection

“…Concerning the mobile phases, buffer solutions could cause problems in the LC system, such as clogging in the pump; an acidic mobile phase is one of the best ways to avoid these problems. The current trend in green chemistry is to avoid the use of buffers [ 34 , 35 , 36 ]. Moreover, isocratic elution was selected with a short running time instead of gradient elution.…”

A Brief Review of Analytical Methods for the Estimation of TTR Kinetic Stabilizers in Pharmaceutical Formulations and Biological Matrices

“…Liquid chromatography (HPLC-UV and LC-MS) is the method of choice for the analysis of TAF plasma levels [46][47][48][49]. DIF is also quantified predominantly by HPLC [50][51][52][53][54][55][56][57][58][59] and LC-MS [60] alone [50][51][52][53]60], in combination with other drugs [54][55][56][57], metabolites [58] or impurities [59]. Most of the analytical methods found in the literature were developed in the 1980s [42, 51-53, 57, 58], some in the 1990s and 2000s [43-45, 50, 54, 55, 60] and two in 2021 [56,59].…”

“…The analytical chromatographic methods for the analysis of DIF described in the literature determined reverse phase liquid chromatography as the preferred fast and reliable method. Both LC-MS [60] procedures and HPLC with UV [42,50,51,54,55,[57][58][59] or FLD detection [52,53,56] adapted for bioassay in plasma [50-53, 56, 60], serum [57,58] or urine [52,53,58] have been developed.…”