摘要
针对以往高效液相色谱法测定谷胱甘肽保留时间过短、易干扰的问题,探索了缓冲盐pH值对还原型和氧化型谷胱甘肽保留时间的影响,建立了一种改良的酵母细胞中谷胱甘肽高效液相色谱分析方法.采用HPLC对酵母粗提物分析,色谱柱为Venusil XBP C18(5μm×10×250mm),甲醇/水为洗脱体系(其中水相中添加庚烷磺酸钠、磷酸-二氢钾,并用磷酸调节pH值),流速1mL/min,柱温25℃,检测波长为203nm.结果表明,还原型谷胱甘肽在0.25~4.00mg/mL质量浓度范围内线性关系良好(R=0.999 7),平均回收率100.49%,RSD为0.52%;氧化型谷胱甘肽在0.25~4.00mg/mL质量浓度范围内线性关系良好(R=0.999 5),平均回收率97.49%,RSD为1.2%.本方法快速简便、灵敏度高,特别是还原型谷胱甘肽保留时间延长、实现与杂峰有效分离.
A modified method for determining glutathione of extracts from yeast using high performance liquid chromatography(HPLC)was established to increase retention time and avoid overlap with other metabolites through adjusting the pH value of water phase of elution system.The extracts of yeast was analyzed by HPLC on a Venusil XBP C-18 column(5μm×10×250 mm)using methanol-water(adding sodium heptanesulfonate,potassium dihydrogen phosphate to the water phase and adjusting the pH with phosphoric acid)as the mobile phase at a flow rate of 1.0 mL/min.The detection wavelength was 203 nm and the temperauture was 25℃.The results show that the reduced glutathione exhibits a good linear relationship in the range of 0.25-4.00 mg/mL;the correlation coefficient is 0.9997;the average recovery is 100.49% and the relative standard deviation(RSD)is 0.52%.Moreover,the oxidized glutathione also exhibits a good linear relationship in the range of 0.25-4.00 mg/mL;the correlation coefficient is 0.9995,the average recovery is 97.49%;and the RSD is 1.2%.The proposed method has the characteristics of rapidity,simpleness and accurateness,especially lengthened retention time and non-interfering separation of glutathione.
作者
陈良立
邓艾平
杜维力
姚鹃
李啸
刘秀继
邓张双
Chen Liangli;Deng Aiping;Du Weili;Yao Juan;Li Xiao;Liu Xiuji;Deng Zhangshuang(Hubei Key Laboratory of Natural Products Research& Development, College of Biological & Pharmaceutical Sciences, China Three Gorges Univ., Yichang 443002, China;, Research & Development Center, Angel Yeast Co., Ltd., Yichang 443002, China;School of Pharmacy, Tongji Medical College of Huazhong Univ. of Science & Technology, Wuhan 430030, China)
出处
《三峡大学学报(自然科学版)》
CAS
北大核心
2018年第4期99-102,共4页
Journal of China Three Gorges University:Natural Sciences
基金
天然产物研究与利用湖北省重点实验室(三峡大学)开放基金项目(2015NP02和2016NP02)
关键词
谷胱甘肽
高效液相色谱法
酵母
glutathione
high performance liquid chromatography
yeast