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植物研究 ›› 2021, Vol. 41 ›› Issue (1): 130-137.doi: 10.7525/j.issn.1673-5102.2021.01.016

• 研究报告 • 上一篇    下一篇

高效液相色谱法测定郁金香鳞茎中3种内源激素

赵杨静, 唐楠(), 唐道城, 张静   

  1. 青海大学高原花卉研究中心,青海省园林植物与观赏园艺重点实验室,西宁 810016
  • 收稿日期:2020-01-10 出版日期:2021-01-20 发布日期:2021-01-05
  • 通讯作者: 唐楠 E-mail:natasha_tn@hotmail.com
  • 作者简介:赵杨静(1995—),女,硕士研究生,主要从事观赏园艺方面的研究。
  • 基金资助:
    国家自然科学基金项目(31660582);青海省“高端创新人才千人计划”;青海省科技厅重点研发与转化计划项目(2018-NK-102)

An Optimized HPLC Procedure for Analyzing Three Endogenous Hormones in Tulip Bulbs

Yang-Jing ZHAO, Nan TANG(), Dao-Cheng TANG, Jing ZHANG   

  1. The Key Laboratory of Landscape Plants and Ornamental Horticulture of Qinghai Province,Plateau Flower Research Center of Qinghai University,Xining 810016
  • Received:2020-01-10 Online:2021-01-20 Published:2021-01-05
  • Contact: Nan TANG E-mail:natasha_tn@hotmail.com
  • About author:ZHAO Yang-Jing(1995—),female,master students,mainly working on ornamental horticulture.
  • Supported by:
    Under the auspices of the National Natural Science Foundation of China(31660582);“Thousand Talents Program” of Qinghai Province;Key Researchand Transformation Project from Science and Technology Department of Qinghai Province(2018-NK-102)

摘要:

利用Agilent高效液相色谱仪,建立了同时测定郁金香鳞茎中GA3、IAA、ABA 3种植物内源激素的高效液相色谱检测方法和异丙醇提取方法。采用外标法,C18反相柱,流动相A(甲醇)∶B(磷酸缓冲液pH=3.5)= 45∶55;流速1 mL·min-1;检测波长0~3.2 min 265 nm,3.0~4.5 min 212 nm,4.5~6.5 min 218 nm,6.5~13.0 min 265 nm;柱温20℃进行测定;以异丙醇提取剂和二氯甲烷低温摇床提纯鳞茎中内源激素。整个过程操作简 单,只需2.0~2.5 h可完成激素提取测定,检测方法线性相关度均在0.995以上,测出限GA3 200 ng·mL-1,IAA 5 ng·mL-1,ABA 20 ng·mL-1;提取方法回收率为84.812%~95.679%,相对标准偏差为6.432%~2.831%。用此方法提取郁金香的内源激素做出的图基线平稳,准确度高,可得到满意的峰形,且操作简单,各环节激素的损失较少。

关键词: 高效液相色谱, 郁金香鳞茎, 内源激素

Abstract:

Agilent HPLC was used to establish a method for simultaneous determination of endogenous hormones GA3, IAA and ABA in tulip bulbs, and an extraction method of isopropanol was established. The external standard method was used for determination, C18 reverse phase column, mobile phase A(methanol)∶B(phosphate buffer pH=3.5)=45∶55; flow rate 1 mL·min-1; detection wavelength 0-3.2 min 265nm, 3.0-4.5 min 212 nm, 4.5-6.5 min 218 nm; 6.5-13.0 min 265 nm; column temperature 20℃ for HPLC determination. Isopropyl alcohol extractant and dichloromethane low temperature shaker to purify the endogenous hormones in the bulb. The whole process was simple to operate, which took only 2.0-2.5 h to complete the hormone purification. The linear correlation of the detection methods was above 0.995. Detection limits of GA3, IAA and ABA were 200 ng·mL-1, 5 ng·mL-1 and 20 ng·mL-1, respectively. Recovery rate of Purification method was 84.812%-95.679%. The relative standard deviation was 6.432%-2.831%. A stable baseline and high accuracy, and a satisfactory peak shape, simple operation, less loss of hormones can be obtained in each link by extracting the endogenous hormone.

Key words: HPLC, tulip bulb, endogenous hormones

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