Mikawy Neven N, Magdy Nancy, Mohamed Marwa H, El-Kosasy Amira M
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Ain Shams University, Abbassia, Cairo, 11566, Egypt.
Sci Rep. 2025 Feb 4;15(1):4252. doi: 10.1038/s41598-024-82387-7.
A combination of three co-administrated drugs, such as azelastine hydrochloride (AZT), fluticasone propionate (FP), and oxymetazoline (OXY), is more effective than single therapy for the treatment of seasonal allergy and COVID-19. We established an efficient methodology for the determination of those analytes in spiked nasal mucosa and nasopharyngeal swabs from real human samples. A simple and quick protein precipitation method was used for sample extraction, using acetonitrile. RP-HPLC/DAD method was performed using an Exsil 100 ODS C18 (250 × 4.6 mm, 5 μm) column with an acetonitrile: water (70:30 v/v) solvent system at a flow rate of 0.7 mL/min. A photodiode array detector was applied at 240 nm. A good separation of the three proposed analytes with a short run time of 10 min was noted. Our method was validated according to FDA guidelines for bioanalytical validation methods. Calibration curves were linear in nasal mucosa samples at concentration ranges of 8-125, 10-100, and 10-125 µg/mL, with average recoveries ± SD of 101.56%±0.39, 102.45%±0.86, and 104.61%±4.52 for AZT, FP, and OXY; respectively. The results of precision and accuracy are within acceptable limits. According to stability assays, the three analytes under investigation were stable throughout sample preparation, storage, and injection. Our method was applied to real nasopharyngeal swabs. It shows that the results of the swabs were not affected by gender or age. Good recoveries with low % RSD were observed: 99.03% ± 0.75, 100.02% ± 0.94, and 100.94% ± 1.98 for both genders, and 100.45% ± 0.96, 100.69% ± 1.08, and 100.32% ± 1.53 for different ages for AZT, FP, and OXY; respectively. Moreover, the amount of those drugs in the nasal mucosa was observed for seven hours, and a constant concentration with a low% RSD was noted for the first four hours. Therefore, this method can be applied to monitor the therapeutic dose in the nasal mucosa for the determination of those analytes.
三种共同给药的药物，如盐酸氮卓斯汀（AZT）、丙酸氟替卡松（FP）和羟甲唑啉（OXY）联合使用，在治疗季节性过敏和新冠肺炎方面比单一疗法更有效。我们建立了一种高效的方法，用于测定来自真实人体样本的加标鼻黏膜和鼻咽拭子中的这些分析物。采用简单快速的蛋白质沉淀法，使用乙腈进行样品提取。采用RP-HPLC/DAD法，使用Exsil 100 ODS C18（250×4.6 mm，5μm）色谱柱，乙腈：水（70:30 v/v）溶剂系统，流速为0.7 mL/min。在240 nm处应用光电二极管阵列检测器。在10分钟的短运行时间内，三种目标分析物得到了良好的分离。我们的方法根据FDA生物分析验证方法指南进行了验证。在鼻黏膜样品中，校准曲线在8-125、10-100和10-125μg/mL的浓度范围内呈线性，AZT、FP和OXY的平均回收率±SD分别为101.56%±0.39、102.45%±0.86和104.61%±4.52。精密度和准确度结果在可接受范围内。根据稳定性试验，所研究的三种分析物在整个样品制备、储存和进样过程中均稳定。我们的方法应用于真实的鼻咽拭子。结果表明，拭子结果不受性别或年龄的影响。观察到回收率良好，RSD%较低：AZT、FP和OXY在男性和女性中的回收率分别为99.03%±0.75、100.02%±0.94和100.94%±1.98，在不同年龄组中的回收率分别为100.45%±0.96、100.69%±1.08和100.32%±1.53。此外，观察了鼻黏膜中这些药物的含量7小时，在前4小时内浓度恒定，RSD%较低。因此，该方法可用于监测鼻黏膜中的治疗剂量，以测定这些分析物。
