Sutariya Vijaykumar, Wehrung Daniel, Geldenhuys Werner J
Department of Pharmaceutical Sciences, Northeast Ohio Medical University, Rootstown, OH 44272, USA.
J Chromatogr Sci. 2012 Mar;50(3):271-6. doi: 10.1093/chromsci/bmr055.
The objective of this study was the development, optimization, and validation of a novel reverse-phase high-pressure liquid chromatography (RP-HPLC) method for the quantification of reduced glutathione in pharmaceutical formulations utilizing simple UV detection. The separation utilized a C18 column at room temperature and UV absorption was measured at 215 nm. The mobile phase was an isocratic flow of a 50/50 (v/v) mixture of water (pH 7.0) and acetonitrile flowing at 1.0 mL/min. Validation of the method assessed the methods ability in seven categories: linearity, range, limit of detection, limit of quantification, accuracy, precision, and selectivity. Analysis of the system suitability showed acceptable levels of suitability in all categories. Likewise, the method displayed an acceptable degree of linearity (r(2) = 0.9994) over a concentration range of 2.5-60 µg/mL. The detection limit and quantification limit were 0.6 and 1.8 µg/mL respectively. The percent recovery of the method was 98.80-100.79%. Following validation the method was employed in the determination of glutathione in pharmaceutical formulations in the form of a conjugate and a nanoparticle. The proposed method offers a simple, accurate, and inexpensive way to quantify reduced glutathione.
本研究的目的是开发、优化和验证一种新型反相高效液相色谱（RP-HPLC）方法，该方法利用简单的紫外检测对药物制剂中的还原型谷胱甘肽进行定量。分离在室温下使用C18柱，并在215nm处测量紫外吸收。流动相是水（pH 7.0）和乙腈按50/50（v/v）混合的等度流动，流速为1.0 mL/min。该方法的验证评估了其在七个类别中的能力：线性、范围、检测限、定量限、准确度、精密度和选择性。系统适用性分析表明所有类别中的适用性水平均可接受。同样，该方法在2.5-60 µg/mL的浓度范围内显示出可接受的线性程度（r(2) = 0.9994）。检测限和定量限分别为0.6和1.8 µg/mL。该方法的回收率为98.80-100.79%。验证后，该方法用于测定共轭物和纳米颗粒形式的药物制剂中的谷胱甘肽。所提出的方法提供了一种简单、准确且廉价的还原型谷胱甘肽定量方法。
