摘要
建立了同时测定甜叶菊中绿原酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸3种化合物的HPLC分析方法,并测定了3种化合物在根、茎、叶中的含量。色谱柱为Waters C18（4.6 mm×250mm,5μm）,流动相为0.2%醋酸（A）和甲醇（B）,流速：1 mL/min,检测波长：327 nm,柱温：40℃,洗脱条件为0~10 min 70%A;10~15 min 70%A-45%A;15~25 min 45%A,25~26 min 45%A-70%A;26~30 min 70%A。在此条件下,绿原酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸的线性范围分别为9.50~285.00、9.25~277.50、9.75~292。50μg/mL且在此范围内线性关系良好（R2〉0.999）,加标回收率介于96.42%~101.18%。从乙醇体积分数、料液比和超声提取时间3因素优化绿原酸提取条件,最佳条件为70%乙醇在料液比1：25的条件下超声提取60 min。对甜叶菊根、茎、叶中上述3种化合物含量进行分析,结果显示,甜叶菊根、茎、叶中绿原酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸的含量范围分别为0.59~7.56、2.43~23.92、1.12~21.21 mg/g。该方法准确度高,重复性好,可用于甜叶菊中3种绿原酸类化合物的同时测定。
A HPLC method was developed for simultaneous determination of three caffeoylquinic acids namely chorogenic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Stevia rebaudianum Bertoni. The content of the three compounds in root, stem and leave were determined by this method. A Waters C18 column（4.6 mm×250 mm, 5 μm） was adopted. The mobile phase consisted of 2% acetic acid（A） and methanol（B） with gradient program of 70%（A） in 0~10 min, 70%~45%（A） in 10~15 min, 45%（A） in 15~25 min, 45%~70%（A） in 25~26 min, 70%（A） in 26~30 min. The flow rate is 1.0 mL/min. The Column temperature was set at 40 ℃, and the detection wavelength was 327 nm. The calibration curves were linear in the ranges of 9.50~285.00, 9.25~277.50, 9.75~292.50 μg/mL for chorogenic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid（R2〉0.999）. The recovery ranged from 96.42% to 101.18%. Three critical extraction parameters namely ethanol concentration, liquid ratio and ultrasonic time were optimized, the best conditions were 1：25 liquid ratio with 70% ethanol ultrasonic extraction for 60 min. Root, stem and leave of Stevia rebaudianum Bertoni were determined by this method. The content of chorogenic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in root, stem and leave of Stevia rebaudianum Bertoni was 0.59~7.56 mg/g, 2.43~23.92 mg/g and 1.12~21.21 mg/g respectively. The determination method was accurate and repeatable, and therefore can be used for simultaneous determination of three caffeoylquinic acid in leaves of Stevia rebaudianum Bertoni.
出处
《食品科技》
CAS
北大核心
2014年第8期276-280,共5页
Food Science and Technology
基金
国家自然科学基金项目(31260368)
江西省人才项目
江西省科技支撑计划项目(2010BNB00503)
关键词
高效液相色谱法
甜叶菊
绿原酸
HPLC
Stevia rebaudianum Bertoni
caffeoylquinic acid
