一种新型的质量设计辅助HPLC-DAD方法同时定量色氨酸、色氨酸和伏立康唑，用于真菌感染的早期诊断和预后，解码群体感应现象

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2025-03-25 DOI:10.1016/j.jchromb.2025.124571

Ahmed M. Saleh , Ola A. Saleh , Rabeay Y.A. Hassan , Amr M. Badawey , Hoda M. Marzouk

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引用次数: 0

摘要

本文首次引入了一种综合分析方法，利用反相高效液相色谱与二极管阵列检测（HPLC-DAD）在单一平台内同时定量代谢前体（色氨酸）、真菌感染群体感应生物标志物（色氨酸）和抗真菌药物（伏立康唑）。该方法采用独特的五氟化结构的Pursuit PFP柱，流动相为甲醇：水（60:40,v/v），流速为1.0 mL/min，检测波长为254.0 nm。采用分析质量设计（AQbD）方法学，结合全因子设计进行最佳方法开发。根据ICH指南进行验证，证明所有目标分析物具有良好的线性（2.0-60.0 μg/mL），同时具有高精度、准确性和系统适用性。此外，当应用于复杂基质时，该方法证明了鲁棒性和通用性，包括加标人血清和药物片剂配方。值得注意的是绿色和白色化学原理的整合，以评估方法的可持续性，代表了分析技术发展的重大进步。分别用AGREE、ComplexGAPI指数、BAGI和RGB 12工具评价绿、蓝、白。这种创新的分析平台为真菌感染的早期检测和实时治疗监测提供了强大的工具。通过同时分析代谢标记物、群体感应特异性生物标记物和抗真菌剂，该方法推进了个性化医疗。它为真菌感染的个性化管理提供了一种新颖、高效和可持续的解决方案，增强了诊断和治疗策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

A novel quality-by-design assisted HPLC-DAD method for the simultaneous quantification of tryptophan, tryptophol, and voriconazole for early diagnosis and prognosis of fungal infections decoding quorum sensing phenomenon

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For the first time, a comprehensive analytical approach is introduced that simultaneously quantifies a metabolic precursor (tryptophan), a quorum-sensing biomarker for fungal infections (tryptophol), and an antifungal drug (voriconazole) within a single platform using reversed-phase high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). The method utilizes a Pursuit PFP column featuring a unique pentafluorinated structure, with a mobile phase of methanol: water (60:40, v/v), a flow rate of 1.0 mL/min, and a detection wavelength set at 254.0 nm. An Analytical Quality-by-Design (AQbD) methodology was employed, incorporating a full factorial design for optimal method development. Validation was performed in accordance with ICH guidelines, demonstrating exceptional linearity (2.0–60.0 μg/mL) for all target analytes, along with high precision, accuracy, and system suitability. Furthermore, the method proved robust and versatile when applied to complex matrices, including spiked human serum and pharmaceutical tablet formulations. Noteworthy is the integration of green and white chemistry principles for evaluating the method's sustainability, representing a significant advancement in analytical technique development. Assessment of greenness, blueness, and whiteness with AGREE, ComplexGAPI index, BAGI and RGB 12 Tools, respectively. This innovative analytical platform provides a powerful tool for the early detection and real-time therapeutic monitoring of fungal infections. By enabling the simultaneous analysis of a metabolic marker, a quorum-sensing specific biomarker, and an antifungal agent, the method advances personalized medicine. It offers a novel, efficient, and sustainable solution for the personalized management of fungal infections, enhancing both diagnostic and therapeutic strategies.

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.