食品科学

• 分析检测 • 上一篇    下一篇

样品前处理对检测柑橘中5-甲基四氢叶酸的影响

郭丽琼,吴厚玖,谈安群,马亚琴,焦晓磊   

  1. 1.西南大学柑橘研究所,重庆 400712;2.西南大学食品科学学院,重庆 400715
  • 出版日期:2013-12-25 发布日期:2014-01-03

Effect of Pretreatments on the Measurement of 5-Methyltetrahydrofolate in Citrus

GUO Li-qiong,WU Hou-jiu,TAN An-qun,MA Ya-qin,JIAO Xiao-lei   

  1. 1. Citrus Research Institute, Southwest University, Chongqing 400712, China;
    2. College of Food Science, Southwest University, Chongqing 400715, China
  • Online:2013-12-25 Published:2014-01-03

摘要:

5-甲基四氢叶酸是叶酸的1种衍生物,是叶酸在柑橘中存在的最主要形式。由于其不稳定及柑橘果实成分复杂,导致叶酸的检测较为困难。要准确测定柑橘中5-甲基四氢叶酸的含量,柑橘样品的前处理非常重要。采用高效液相荧光检测法,考察不同的前处理方法对柑橘中5-甲基四氢叶酸检出结果的影响。结果表明:柑橘类样品果肉、果皮、囊衣3g,种子2g匀浆后加入20mL磷酸缓冲液,调解pH值为6.8,沸水浴处理15min(1次水解),冷却离心取上清液定容至25mL后取3mL,添加0.5mL大鼠血清、37℃振摇温育20h时,可得到最佳检测结果。

关键词: 前处理, 柑橘, 5-甲基四氢叶酸, 检测

Abstract:

The main form of folate, as a polyglutamate derivative of folate, in citrus is 5-methyltetrahydrofolate (5-MTHF).
The content of 5-MTHF is difficult to measure due to its instability and the complex matrix in citrus. Pre-treatment of
samples is very important for determining 5-MTHF in citrus accurately. In this work, the effect of different pre-treatment
methods on the extraction efficiency of 5-MTHF was explored using high performance liquid chromatography-fluorescence
detection (RP-HPLC-FLD). The results showed that the best hydrolysis conditions for citrus samples were found as follows:
homogenization of samples (3 g of pulp, exocarp plus mesocarp, endocarp or 2 g of seeds) and dilution to 20 mL with
phosphate buffer and adjustment of pH to 6.8, heating in a boiling water bath for 15 min (single hydrolysis), dilution of
supernatant to 25 mL after cooling down and centrifugation, addition of 0.5 mL of rat serum to 3 mL of the diluted sample,
and incubation at 37 ℃ for 20 h with an adjusted pH of 6.8 while shaking.

Key words: pre-treatment, citrus, 5-methyltetrahydrofolate (5-MTHF), detetermination