色谱

色谱

• 研究论文 • 上一篇    下一篇

反相高效液相色谱法测定烤烟叶片发育过程中的类胡萝卜素类物质

刘国顺1,韦凤杰1,王芳1,李亚娟1,郭桥燕1,黄晓书2

  

  1. 1.National Tobacco Cultivation, Physiology and Biochemistry Research Center, Henan Agricultural University, Zhengzhou 450002, China; 2.Key Discipline and Open Laboratory of Agricultural Biological and Engineering Technique, Henan Institute of Higher Learning, Henan Agriculture University, Zhengzhou 450002, China
  • 收稿日期:2005-03-07 修回日期:2005-11-21 出版日期:2006-03-30 发布日期:1987-06-25
  • 通讯作者: 刘国顺

Determination of Carotenoids in Flue-Cured Tobacco Leaves During its Growth by Reversed-Phase High Performance Liquid Chromatography

LIU Guoshun1, WEI Fengjie1, WANG Fang1, LI Yajuan1, GUO Qiaoyan1, HUANG Xiaoshu2

  

  1. 1.National Tobacco Cultivation, Physiology and Biochemistry Research Center, Henan Agricultural University, Zhengzhou 450002, China; 2.Key Discipline and Open Laboratory of Agricultural Biological and Engineering Technique, Henan Institute of Higher Learning, Henan Agriculture University, Zhengzhou 450002, China
  • Received:2005-03-07 Revised:2005-11-21 Online:2006-03-30 Published:1987-06-25
  • Contact: LIU Guoshun

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摘要：

建立了采用反相高效液相色谱测定烤烟叶片中类胡萝卜素的方法。烤烟叶片先用含0.1%丁基羟基甲苯（BHT）的90%丙酮水溶液萃取,然后加入0.1 g醋酸铅,于4 ℃下以10000 r/min离心5 min以去除蛋白质。色谱柱为 C18反相柱(3.9 mm i.d.×150 mm, 5 μm)。流动相:A，甲醇-异丙醇(体积比为1∶1）；B，超纯水。洗脱程序:0~10 min，70%A+30%B；10~17 min，100%A；17~30 min(90%A+10%B)。流速：0.5 mL/min。进样量:10 μL。检测波长:450 nm。该方法简化了样品的前处理过程,4种类胡萝卜素物质的加标回收率为91.77%~97.42%,相对标准偏差为
3.46%~0.98%。用该方法研究了烤烟发育过程中类胡萝卜素含量的变化规律,获得了与文献较为一致的结果。

关键词: 反相高效液相色谱法, 烤烟叶片 , 类胡萝卜素

Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method for the determination of carotenoids in flue-cured tobacco leaves was developed. Carotenoids were extracted from flue-cured tobacco leaves by acetone-water (90∶10, v/v) solution containing 0.1% butylated hydroxytoluene (BHT). Plant proteins were eliminated by adding 0.1 g Pb(Ac)2 and by centrifugation (10000 r/min) for 5 min at 4 ℃. Lutein, β-carotene, neoxanthin, violaxanthin and other plant pigments were separated on a reversed-phase C18 column (3.9 mm i.d.×150 mm, 5 μm), with a mobile phase of (A) methanol-isopropyl alcohol (1∶1, v/v) and (B) water using a gradient elution at a flow rate of 0.5 mL/min. The optimum elution gradient was as follows: 0-10 min, 70%A+30%B; 10-17 min, 100%A; 17-30 min, 90%A＋10%B. The recoveries of carotenoids in flue-cured tobacco leaves were 91.77%-97.42%, and relative standard deviations were 3.46%-0.98%. This method was applied to determine carotenoids in flue-cured tobacco leaves during its growth with satisfactory results.

Key words: flue-cured tobacco leaves , carotenoids, reversed-phase high performance liquid chromatography (RP-HPLC)