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Bulletin of Botanical Research ›› 2021, Vol. 41 ›› Issue (1): 130-137.doi: 10.7525/j.issn.1673-5102.2021.01.016

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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)

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

CLC Number: