Khan Nasr Ullah, Iqbal Muhammad, Khan Barkat Ali
Gomal Center of Pharmaceutical Sciences, Faculty of Pharmacy, Gomal University, Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan.
PLoS One. 2025 Apr 15;20(4):e0321474. doi: 10.1371/journal.pone.0321474. eCollection 2025.
The implementation of quality by design offer quality and safety product to patients, efficient processes for continuous improvements to manufacturers, negligible amount of batch failures and robust product quality attributes. This study was designed to establish a simple, specific, precise, and accurate reverse phase-high performance liquid chromatographic (RP-HPLC) method for the quantitative determination of remdesivir and favipiravir in Capsule dosage forms. The RP-HPLC method was performed on a Kromasil 100A C18 column (250 mm x 4.6 mm, 5 μm particle size) with a mobile phase containing (50 mL of acetonitrile, 350 mL methanol, 100ml of water and 0.5 mL of Phosphoric acid). The flow rate was 1.0 mL/min. The ultraviolet (UV) detection wavelength was 300 nm, and the column temperature was set at ambient. Linearity and range stock solutions were prepared as 50% to 150%. The calibration curves showed a good linear response ranged from 0.02 to 0.06 mg/ml (r=1.0000) for favipiravir and 0.022 to 0.066 mg/ml (r=1.0000) for remdesivir, and the average recoveries were 99.9% for favipiravir and 99.8% for remdesivir in assay test. The retention time of favipiravir was RT 11.5 minutes and remdesivir 20.95 minutes, tailing factor was not more than 1.5 and resolution of more than 2.0 respectively. The extended run time was supported by high concentration of favipiravir and uneven peak behavior of remdesivir in single run, which disturb resolution, retention times, injection repeatability. Reducing runtime on HPLC will lead to coelution of peaks, poor resolution, and loss of sensitivity during degradation profiling. This issue may be overcome with future UHPLC technique. The method was validated in accordance to International Council for Harmonization of Technical Requirements of Pharmaceuticals for Human Use (ICH) guidelines. The limit of detection (LOD) for Favipiravir was 0.104mg/ml and Remdesivir was 0.052mg/ml and limit of quantification (LOQ) for Favipiravir was 0.316mg/ml and Remdesivir was 0.158mg/ml respectively. In stress conditions, this product undergoes degradation and was considered sensitive to Acid, Alkali, Oxidation, reduction, hydrolysis, high temperature and humidity. The assay methods was simple, rapid, sensitive, repeatable, eco-friendly, stability indicating and can be used cost-effectively for the testing of these two drug substances in capsule dosage form. Due to the emergent need of antivirals for flu management, this coloaded combination was considered as dire need of society. Also, we reported that the product must be stored under ambient temperature, low humidity, and protected from light exposure.
质量源于设计的实施为患者提供了质量可靠且安全的产品,为制造商提供了持续改进的高效流程,使批次失败数量可忽略不计,并具备稳健的产品质量属性。本研究旨在建立一种简单、特异、精密且准确的反相高效液相色谱(RP-HPLC)法,用于定量测定胶囊剂型中瑞德西韦和法匹拉韦的含量。RP-HPLC法在Kromasil 100A C18柱(250 mm×4.6 mm,粒径5μm)上进行,流动相包含(50 mL乙腈、350 mL甲醇、100 mL水和0.5 mL磷酸)。流速为1.0 mL/min。紫外(UV)检测波长为300 nm,柱温设定为室温。线性范围储备溶液的配制浓度为50%至150%。校准曲线显示,法匹拉韦在0.02至0.06 mg/ml范围内呈良好线性响应(r = 1.0000),瑞德西韦在0.022至0.066 mg/ml范围内呈良好线性响应(r = 1.0000),在含量测定试验中,法匹拉韦的平均回收率为99.9%,瑞德西韦的平均回收率为99.8%。法匹拉韦的保留时间为11.5分钟,瑞德西韦的保留时间为20.95分钟,拖尾因子分别不超过1.5和分离度大于2.0。单次进样中,高浓度的法匹拉韦和瑞德西韦峰形不均一,导致运行时间延长,干扰了分离度、保留时间和进样重复性。在降解图谱分析过程中,减少HPLC的运行时间会导致峰共洗脱、分离度差和灵敏度损失。未来的超高效液相色谱(UHPLC)技术可能会克服这个问题。该方法按照国际人用药品注册技术协调会(ICH)指南进行了验证。法匹拉韦的检测限(LOD)为0.104 mg/ml,瑞德西韦的检测限为0.052 mg/ml,法匹拉韦的定量限(LOQ)为0.316 mg/ml,瑞德西韦的定量限为0.158 mg/ml。在强制条件下,该产品会发生降解,且被认为对酸、碱、氧化、还原、水解、高温及湿度敏感。该含量测定方法简单、快速、灵敏、可重复、环保、具有稳定性指示作用,且可经济高效地用于检测胶囊剂型中的这两种药物。由于对抗病毒药物治疗流感的迫切需求,这种联合载药组合被视为社会的迫切需求。此外,我们报告该产品必须在室温、低湿度下储存,并避免光照。