Sri Ramachandra Institute of Higher Education and Research (DU), Department of Pharmaceutical Chemistry, Sri Ramachandra Faculty of Pharmacy, Chennai, Tamil Nadu 600116, India.
SRM College of Pharmacy, SRM Institute of Science and Technology, Department of Pharmaceutical Analysis, Kattankulathur, Tamil Nadu 603203, India.
J AOAC Int. 2024 Jan 4;107(1):14-21. doi: 10.1093/jaoacint/qsad105.
Glutathione, silybin, and curcumin are well-known potential antioxidants that are recommended as adjuvant therapy in cancer treatment.
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.
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.
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.
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.
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.
谷胱甘肽、水飞蓟宾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。
基于分析质量源于设计（AQbD）和绿色分析化学的原则，建立了一种简单、稳健且环境友好的 HPLC 法，用于同时测定原料药和制剂中的谷胱甘肽、水飞蓟宾和姜黄素。
采用 Agilent Eclipse XDB C18（150mm×4.6mm id，3.5μm）柱，以乙醇-水 pH 6.7（含 0.1%，v/v 正磷酸）为流动相，梯度洗脱，流速 1.07mL/min，PDA 检测波长 215nm。采用石川图进行风险评估，确定关键方法变量，并通过实验设计（DoE）软件的中心复合设计对 HPLC 技术的实验条件进行多变量优化。
分离在 15min 内完成，谷胱甘肽、水飞蓟宾和姜黄素的保留时间分别为 3.3、4.9 和 7.3min。谷胱甘肽在 3.75-26.25μg/mL、水飞蓟宾在 62.50-437.50μg/mL、姜黄素在 12.5-87.50μg/mL 范围内呈线性关系。该方法经 ICH 指南 Q2（R1）验证，所有参数均在规定范围内。
该方法简单、准确、稳健，可用于常规分析，并且根据 AGREE、绿色分析程序指数和分析生态规模工具，被认为是一种更绿色的方法。
首次采用色谱技术同时评价所选抗氧化剂在原料药和制剂中的应用，采用绿色溶剂。采用 PDA 检测器研究了所有三种化合物的峰纯度。通过采用 AQbD 要素和工具，减少了时间、成本和溶剂的浪费。完全采用环境可持续的安全溶剂。
