Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
Molecules. 2023 Jul 5;28(13):5221. doi: 10.3390/molecules28135221.
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.
在这项研究中，首次开发并验证了一种新的绿色微量荧光分析（MW-SFA）方法，具有高通量，用于测定药物制剂和血浆中的三种选择性 5-羟色胺再摄取抑制剂（SSRIs）。这些 SSRIs 是氟西汀（FLX）、氟伏沙明（FXM）和帕罗西汀（PXT），它们是常用于治疗抑郁症的处方药物。MW-SFA 基于 SSRIs 在碱性介质中与 4-氯-7-硝基苯并-2-氧杂-1,3-二唑（NBD-Cl）的缩合反应，形成高荧光衍生物。MW-SFA 程序在平底白色不透明 96 微孔板中进行，使用微孔板读数器在其最大激发和发射波长处测量荧光信号。校准曲线在 35-800ng/mL 范围内具有良好的相关系数（0.9992-0.9995），相对荧光强度（RFI）与 SSRIs 浓度之间存在相关性。检测限范围在 11-25ng/mL 之间，精密度和准确度令人满意。所提出的 MW-SFA 成功应用于 SSRIs 药物制剂的分析。MW-SFA 测定结果与药典测定结果的统计分析表明，两种测定方法在准确度和精密度方面无显著差异。该方法的应用扩展到成功分析血浆样品中的 SSRIs。使用三种不同的度量工具证实了测定的绿色性。该测定具有高通量特性，能够在短时间内灵敏地同时分析许多样品。该测定方法对于药物质量控制单位和临床实验室中 SSRIs 的快速常规应用具有重要价值。
