Attia Khalid A M, El-Olemy Ahmed, Ramzy Sherif, Abdelazim Ahmed H, Hasan Mohamed A, Omar Mahmoud K M, Shahin Mohamed
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
Spectrochim Acta A Mol Biomol Spectrosc. 2021 Jan 5;244:118871. doi: 10.1016/j.saa.2020.118871. Epub 2020 Aug 22.
Lesinurad and allopurinol combination is newly FDA approved for treatment of patients suffering from hyperuricemia associated with uncontrolled gout. In the present work, two different highly sensitive, selective and accurate fluorescence spectroscopic methods were developed for quantitative analysis of lesinurad and allopurinol in their pharmaceutical dosage form without any tedious operation procedure. Lesinurad was quantitatively analyzed based on its unique native fluorescence nature. Lesinurad fluorescence emission was quantitatively determined at 343 nm after excitation at 288 nm without any interference from allopurinol. Allopurinol, has free terminal secondary amino group, reacted with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBDCl) through nucleophilic substitution mechanism forming highly fluorescent dark yellow fluorophore. Allopurinol was quantitavely analyzed based on measurement the emission fluorescence intensity of the fluorescent dark yellow fluorophore at 535 nm after excitation at 465 nm. Different parameters which affect the described methods of the studied drugs were carefully checked and optimized. Calibration graphs were found to be linear over the concentration range of 0.25-4.0 μg/mL for lesinurad and 0.2-20 μg/mL for allopurinol. The proposed methods were successfully applied for the quantitative analysis of the two drugs in Duzallo® pharmaceutical dosage form and spiked human plasma.
雷西纳德与别嘌醇的组合已获美国食品药品监督管理局（FDA）新批准，用于治疗与痛风控制不佳相关的高尿酸血症患者。在本研究中，开发了两种不同的高灵敏度、高选择性且准确的荧光光谱法，用于定量分析其药物剂型中的雷西纳德和别嘌醇，无需任何繁琐的操作步骤。雷西纳德基于其独特的天然荧光性质进行定量分析。在288nm激发后，于343nm处定量测定雷西纳德的荧光发射，不受别嘌醇的任何干扰。别嘌醇具有游离的末端仲氨基，通过亲核取代机制与4-氯-7-硝基苯并-2-恶唑-1,3-二氮杂茂（NBDCl）反应，形成高荧光的暗黄色荧光团。基于在465nm激发后，测量535nm处暗黄色荧光团的发射荧光强度，对别嘌醇进行定量分析。仔细检查并优化了影响所研究药物上述方法的不同参数。发现校准曲线在雷西纳德浓度范围为0.25 - 4.0μg/mL和别嘌醇浓度范围为0.2 - 20μg/mL内呈线性。所提出的方法成功应用于Duzallo®药物剂型和加标人血浆中两种药物的定量分析。
