To analyze Hesperidin (HES) in its pure form and in supplement formulations, a new spectrophotometric method was developed using a flow injection system (FI) as an integrated, semi-automated technique for quantitative analysis. An orange-colored product was produced during the reaction of HES with diazotized Procaine penicillin (PRP), and was spectrophotometrically detectable at 434nm wavelength (HES–N=N–PRP). The optimum conditions for the Flow injection method were established after evaluating all relevant chemical and physical parameters. The linearity range (LR), the correlation coefficient(r), limit of detection (LOD), and limit of quantification (LOQ) were obtained at 5 -100 μg mL-1, 0.9936, 0.679 μg mL-1, and 1.86 μg mL-1, respectively, for 79 samples per hour. The Azo bond (R-N=N-R) was obtained between the HES ring (nucleophilic) and diazotized PRP (electrophile). So, the reaction creates a highly conjugated chromophore that forms an intensely colored azo compound dye. The procedure demonstrated successful applicability for HES quantification in pharmaceutical formulations. Finally, the environmental sustainability of the developed method was assessed using complex MoGAPI and AGREE Preparation techniques. The evaluation confirmed that the approach is both environmentally friendly and aligned with green analytical principles.
Abed et al. (Sun,) studied this question.