A Novel quality-by-design optimized spectrofluorimetric approach for estimation of the oral synthetic adiponectin receptor agonist AdipoRon in different biological matrices: Compliance with greenness metrics

Abstract

The current study introduces the first analytical approach for the quantitative analysis of adipoRon, a synthetic adiponectin receptor agonist, using a facile, rapid, sensitive, and green spectrofluorimetric method. Nitrogen-doped carbon quantum dots (N-CQDs) were synthesized by adopting a fast microwave-assisted procedure using accessible and natural substrates (orange juice and urea). The suggested synthetic method produced CQDs with narrow particle size distribution (2–10 nm), high fluorescence stability, and good quantum yield (29.3%). The produced fluorescent QDs were used as fluorescent sensors and were quantitatively quenched by increasing adipoRon concentrations. A two-level full factorial design (22 FFD) was used to identify the most critical experimental factors affecting the relative fluorescence intensity of N-CQDs. The fluorescence quenching of N-CQDs by the drug was measured at 417 nm after excitation at 325 nm. Under the optimized conditions, the quenching of N-CQDs' fluorescent intensity displayed acceptable linearity with the adipoRon concentration range of 5.0–20.0 μg/mL with limits of quantitation and detection of 0.843 μg/mL and 0.278 μg/mL, respectively. Owing to the high sensitivity, the suggested method was efficiently applied for adipoRon estimation in different biological matrices, including spiked human plasma, real rat plasma, and liver homogenate samples with high % recoveries (87.41–103.17%) and low %RSD values (<2.87%). The method's greenness was evaluated using two metric tools, the complementary green analytical procedure index (ComplexGAPI) and the Analytical GREENNESS metric approach (AGREE), and the results showed that the method was excellent green. The proposed methodology was validated according to the ICHQ2 (R1) recommendations.