An RP-HPLC Method for the Simultaneous Analysis of Selected Antiviral Drugs

IF 1.2 4区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Chromatographia Pub Date : 2025-02-05 DOI:10.1007/s10337-025-04386-8

Mehmet Emre Özdemirhan, Gürkan Özen, Emirhan Nemutlu

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引用次数: 0

Abstract

A rapid and efficient reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous determination of six antiviral drugs: emtricitabine (EMT), entecavir (ENT), favipiravir (FAV), ganciclovir (GAN), valaciclovir (VAL), and valganciclovir (VLG). Chromatographic separation was achieved on an ExsilTM Mono 100 C18 column using a mobile phase of 50 mM phosphate buffer (pH 4): acetonitrile (90:10, v/v) at a flow rate of 0.5 mL/min. UV detection was performed at 215 nm. The method was validated according to ICH Q2(R2) guidelines, demonstrating excellent linearity (R² > 0.9938) across the concentration ranges of 1.0–150 μg/mL for EMT, ENT and VLG, and 5.0–200 μg/mL for FAV, GAN and VAL. The LOD and LOQ values ranged from 0.02 to 0.48 μg/mL and 0.05 to 1.45 μg/mL, respectively. Intra-day and inter-day precision, expressed as relative standard deviation (RSD), were less than 1.82%, while accuracy, expressed as relative error (RE), was within ± 1.34%. Recovery studies yielded values ranging from 98.12% to 101.60%, further confirming method accuracy. The method proved to be selective, robust, and suitable for the quantitative analysis of these antiviral drugs in pharmaceutical formulations, with results obtained for commercially available tablets demonstrating excellent agreement with labeled contents. Notably, stability studies revealed the susceptibility of EMT, ENT, and FAV to degradation under refrigerated storage, highlighting the need for timely analysis of these compounds. In addition, the greenness, blueness, and whiteness of the method were evaluated, and the developed method performs strongly across environmental (AGREE score: 0.79), practical (BAGI index: 85), and sustainability (whiteness value: 74.7) criteria, reinforcing its viability as a sustainable approach for antiviral analysis. This validated RP-HPLC method provides a reliable and efficient tool for the quality control of these essential antiviral medications.

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反相高效液相色谱法同时分析选定的抗病毒药物

建立了一种快速高效的反相高效液相色谱（RP-HPLC）方法，用于同时测定恩曲他滨（EMT）、恩替卡韦（ENT）、法匹拉韦（FAV）、更昔洛韦（GAN）、伐昔洛韦（VAL）和伐更昔洛韦（VLG） 6种抗病毒药物。色谱分离采用ExsilTM Mono 100 C18色谱柱，流动相为50 mM磷酸盐缓冲液（pH 4）：乙腈（90:10,v/v），流速为0.5 mL/min。在215 nm处进行紫外检测。结果表明：EMT、ENT和VLG在1.0 ~ 150 μg/mL范围内，FAV、GAN和VAL在5.0 ~ 200 μg/mL范围内呈良好的线性关系（R2 > 0.9938）， LOD和LOQ分别在0.02 ~ 0.48 μg/mL和0.05 ~ 1.45 μg/mL范围内。以相对标准偏差（RSD）表示的日内和日内精密度小于1.82%，以相对误差（RE）表示的准确度在±1.34%以内。回收率在98.12% ~ 101.60%之间，进一步证实了方法的准确性。该方法被证明是选择性的，稳健的，适用于药物制剂中这些抗病毒药物的定量分析，对市售片剂的结果显示与标签内容物非常吻合。值得注意的是，稳定性研究揭示了EMT、ENT和FAV在冷藏条件下对降解的敏感性，强调了对这些化合物进行及时分析的必要性。此外，对该方法的绿度、蓝度和白度进行了评估，发现该方法在环境（AGREE得分：0.79）、实用（BAGI指数：85）和可持续性（白度值：74.7）标准上表现良好，增强了其作为抗病毒分析的可持续方法的可行性。该方法为这些重要抗病毒药物的质量控制提供了可靠、高效的工具。

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来源期刊

Chromatographia 化学-分析化学

CiteScore

3.40

自引率

5.90%

发文量

103

审稿时长

2.2 months

期刊介绍： Separation sciences, in all their various forms such as chromatography, field-flow fractionation, and electrophoresis, provide some of the most powerful techniques in analytical chemistry and are applied within a number of important application areas, including archaeology, biotechnology, clinical, environmental, food, medical, petroleum, pharmaceutical, polymer and biopolymer research. Beyond serving analytical purposes, separation techniques are also used for preparative and process-scale applications. The scope and power of separation sciences is significantly extended by combination with spectroscopic detection methods (e.g., laser-based approaches, nuclear-magnetic resonance, Raman, chemiluminescence) and particularly, mass spectrometry, to create hyphenated techniques. In addition to exciting new developments in chromatography, such as ultra high-pressure systems, multidimensional separations, and high-temperature approaches, there have also been great advances in hybrid methods combining chromatography and electro-based separations, especially on the micro- and nanoscale. Integrated biological procedures (e.g., enzymatic, immunological, receptor-based assays) can also be part of the overall analytical process.