Green HPLC Method for Simultaneous Analysis of Three Natural Antioxidants by Analytical Quality by Design.

IF 1.7 4区 农林科学 Q3 CHEMISTRY, ANALYTICAL Journal of AOAC International Pub Date : 2024-01-04 DOI:10.1093/jaoacint/qsad105
Ramya Jonnalagadda, Seetharaman Rathinam, Krishnaveni Nagappan, Vinodhini Chandrasekar
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Abstract

Background: Glutathione, silybin, and curcumin are well-known potential antioxidants that are recommended as adjuvant therapy in cancer treatment.

Objective: Based on the principles of Analytical Quality by Design (AQbD) and green analytical chemistry, a simple, robust, and environmentally benign HPLC method for the simultaneous estimation of glutathione, silybin, and curcumin in bulk and formulation was performed.

Method: Elution was achieved by an Agilent Eclipse XDB C18 (150 mm × 4.6 mm id, 3.5 μm) column using a gradient mobile phase composed of ethanol-water pH 6.7 (with 0.1%, v/v orthophosphoric acid) and 1.07 mL/min flow rate with PDA detection at 215 nm. Critical method variables were identified by risk assessment using an Ishikawa diagram, and multivariate optimization of the experimental conditions for the HPLC technique was accomplished by central composite design using design of experiments (DoE) software.

Results: The separation was achieved within 15 min, where the retention time of glutathione, silybin, and curcumin were 3.3, 4.9, and 7.3 min, respectively. The standard curve was linear in the range of 3.75-26.25 µg/mL for glutathione, 62.50-437.50 µg/mL for silybin, and 12.5-87.50 µg/mL for curcumin. The developed method was validated as per ICH guidelines Q2 (R1), and all the parameters are within specified limits.

Conclusions: The proposed method is simple, precise, and robust, which can be employed for routine analysis and also concluded to be a greener approach according to AGREE, Green Analytical Procedure Index, and analytical eco-scale tools.

Highlights: The chosen antioxidants were evaluated for the very first time simultaneously using the chromatographic technique in bulk and dosage forms employing green solvents. The peak purity of all three compounds was studied using a PDA detector. Wastage was reduced in terms of time, cost, and solvents by employing AQbD elements and tools. Complete application of environmentally sustainable safe solvents were employed.

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通过 "分析质量源于设计 "实现同时分析三种天然抗氧化剂的绿色高效液相色谱法。
背景:谷胱甘肽、水飞蓟素和姜黄素是众所周知的潜在抗氧化剂,被推荐作为癌症治疗的辅助疗法:基于设计分析质量(AQbD)和绿色分析化学的原则,建立了一种简单、稳健、对环境无害的高效液相色谱法,用于同时测定散装和制剂中的谷胱甘肽、水飞蓟素和姜黄素:采用Agilent Eclipse XDB C18 (150 mm × 4.6 mm id, 3.5 μm)色谱柱,以pH 6.7的乙醇-水(含0.1% v/v正磷酸)为流动相梯度洗脱,流速为1.07 mL/min,在215 nm波长下进行PDA检测。通过使用石川图进行风险评估,确定了关键的方法变量,并使用实验设计(DoE)软件通过中心复合设计对高效液相色谱技术的实验条件进行了多变量优化:谷胱甘肽、水飞蓟素和姜黄素在 15 分钟内实现分离,保留时间分别为 3.3、4.9 和 7.3 分钟。谷胱甘肽、水飞蓟素和姜黄素在3.75-26.25微克/毫升、62.50-437.50微克/毫升和12.5-87.50微克/毫升范围内线性关系良好。根据 ICH 指南 Q2 (R1),对所开发的方法进行了验证,所有参数均在规定的范围内:结论:所提出的方法简单、精确、稳健,可用于常规分析,根据 AGREE、绿色分析程序指数和分析生态尺度工具,该方法是一种更环保的方法:首次使用色谱技术同时评估了所选抗氧化剂在使用绿色溶剂的散装和剂型中的应用。使用 PDA 检测器研究了所有三种化合物的峰纯度。通过使用 AQbD 元素和工具,减少了时间、成本和溶剂方面的浪费。完全采用了环境可持续的安全溶剂。
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来源期刊
Journal of AOAC International
Journal of AOAC International 医学-分析化学
CiteScore
3.10
自引率
12.50%
发文量
144
审稿时长
2.7 months
期刊介绍: The Journal of AOAC INTERNATIONAL publishes the latest in basic and applied research in analytical sciences related to foods, drugs, agriculture, the environment, and more. The Journal is the method researchers'' forum for exchanging information and keeping informed of new technology and techniques pertinent to regulatory agencies and regulated industries.
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