Development of a Green Microwell Spectrofluorimetric Assay with High Analytical Throughput for the Determination of Selective Serotonin Reuptake Inhibitors in Pharmaceutical Dosage Forms and Plasma

Abstract

In this study, a new green microwell spectrofluorimetric assay (MW-SFA) with high throughput was developed and validated, for the first time, for the determination of three selective serotonin reuptake inhibitors (SSRIs) in pharmaceutical dosage forms and plasma. These SSRIs were fluoxetine (FLX), fluvoxamine (FXM), and paroxetine (PXT), which are commonly prescribed drugs for depression treatment. The MW-SFA is based on the condensation reaction of SSRIs with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) in alkaline media to form highly fluorescent derivatives. The MW-SFA procedures were conducted in 96-microwell white opaque assay plates with a flat bottom and the fluorescence signals were measured using a microplate reader at their maximum excitation and emission wavelengths. The calibration curves were generated with good correlation coefficients (0.9992–0.9995) between the relative fluorescence intensity (RFI) and the SSRI concentrations in the range of 35–800 ng/mL. The limits of detection were in the range of 11–25 ng/mL, and the precision and accuracy were satisfactory. The proposed MW-SFA was successfully applied to the analysis of the SSRIs in their pharmaceutical dosage forms. The statistical analysis for the comparison between the MW-SFA assay results and those of pharmacopeial assays showed no significant differences between the assays in terms of their accuracy and precision. The application of the proposed MW-SFA was extended to successfully analyze SSRIs in plasma samples. The greenness of the assay was confirmed using three different metric tools. The assay was characterized with high throughput properties, enabling the sensitive simultaneous analysis of many samples in a short time. This assay is valuable for rapid routine applications in pharmaceutical quality control units and clinical laboratories for the determination of SSRIs.

A facile on–off fluorescence approach for fluvoxamine determination in pharmaceutical tablets; application to content uniformity testing

“…The suggested technique processes several benefits over the spectrofluorimetric techniques that have previously been published. 7–12 The present method incorporate the use of a readily accessible and less expensive reagent (erythrosine B) rather than fluorescamine, which was previously described for FLX measurement. 12 The interaction between the examined medication and erythrosine B was instantaneous at room temperature, which is fairly quick.…”

“…Thus, there was no need to stand for any period of time prior to the measurements. On the other hand, in four methods of the previous techniques, 7–10 the reaction mixtures were heated at high temperatures for long time, in multi-steps procedures. Moreover, the method utilized water as a green solvent so it is more ecologically friendly than other published methods, which use risky organic solvents like acetonitrile.…”

“…Reviewing the literature shows that there are numerous approaches for determining the medication under investigation. These strategies include: spectrophotometry, 3–6 spectrofluorimetry, 7–12 liquid chromatography (HPLC), 13–18 gas chromatography (GC), 19 thin layer chromatography, 20–22 electrochemical methods, 23–25 and capillary electrophoresis. 26 HPLC necessitates the use of expensive instruments and huge quantities of highly pure liquids.…”

“…Meanwhile, spectrofluorimetric methods are more selective due to the presence of two spectra, extremely sensitive, and need a modest instrument as well as very easy sample preparation. Nevertheless, the published spectrofluorimetric procedures for the mentioned medication suffered from certain limitations such as the use of high-priced chemical, 12 damaging organic solvents, 9 high degrees of temperature for extended periods of time, 7–10 or possesses low sensitivity. 11…”

Tuning fluorescence of fexofenadine by switching OFF intramolecular photoinduced electron transfer: Application to development of one‐step green and high throughput microwell spectrofluorimetric assay for analysis of tablets and plasma

“…Furthermore, they are adaptable to several detection techniques, including absorbance, fluorescence, and luminescence, facilitating both quantitative and qualitative assessments and providing a more detailed analysis of the samples in question [25, 26]. Recently, successful microwell spectrophotometric [27] and spectrofluorimetric assays [28] have been described for the determination of different drugs in dosage forms and biological fluids. Compared to spectrophotometric assays, spectrofluorimetric assays offer greater simplicity and higher sensitivity.…”