Determination method of content of endogenous hormones in terminal buds of tomato plants

A technology for endogenous hormones and determination methods, which is applied in the field of crop endogenous hormones in agronomy, can solve the problems of preparing mobile phases and setting high requirements for chromatographic conditions, and achieves the effect of rapid determination.

Active Publication Date: 2015-07-08
怀远县翰邦科技咨询有限公司
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Problems solved by technology

High-performance liquid chromatography has the characteristics of fast, efficient, high-sensitivity, and high-automation, but it has high requirements for preparing mobile phase and setting chromatographic conditions. At present, there is no sample suitable for high-performance liquid chromatography to determine endogenous hormones in tomato terminal buds Methods of pretreatment, mobile phase, and setting chromatographic conditions

Method used

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  • Determination method of content of endogenous hormones in terminal buds of tomato plants
  • Determination method of content of endogenous hormones in terminal buds of tomato plants
  • Determination method of content of endogenous hormones in terminal buds of tomato plants

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Experimental program
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preparation example Construction

[0041] 3) Preparation of standard solution: the standard solution of plant hormones includes 0.1g / L IAA standard solution, 0.001g / L ZT standard solution, 0.001g / L ABA standard solution and 0.01g / L GA 3 Standard solution; prepare the standard solution according to different concentration ratios to prepare a series of standard mixed solutions with concentration gradients, and store at 4°C;

[0042] 4) Examination of specificity: under the chromatographic conditions of step 2), the investigation of specificity includes IAA, ZT, ABA and GA in the tomato terminal bud sample 3 Perform HPLC separation and detection;

[0043] 5) Linear relationship investigation: Measure the IAA, ZT, ABA and GA made by step 3) respectively 3 1mL each standard solution, dilute to the volume with methanol in a 10mL brown volumetric flask, filter, inject and detect, and perform linear regression analysis with the peak areas of the four mixed standards and the corresponding mass concentrations;

[0044] 6...

Embodiment

[0047] 1. Material preparation

[0048] ① Material to be tested

[0049] Samples of pretreated tomato terminal shoots.

[0050] ② Reagent:

[0051] IAA standard product, SIGMA-ALDRICH, product of USA;

[0052] ZT standard product, SIGMA-ALDRICH, product of USA;

[0053] ABA standard product, SIGMA-ALDRICH, product of USA;

[0054] GA 3 Standard product, SIGMA-ALDRICH, product of USA;

[0055] Methanol, chromatographically pure, Sinopharm Chemical Reagent Co., Ltd., Shanghai;

[0056] Glacial acetic acid, analytically pure, Sinopharm Chemical Reagent Co., Ltd., Shanghai.

[0057] ③Instrument:

[0058] 10-1000μL DRAGONLAB adjustable pipette, Shanghai Wandao Instrument Technology Co., Ltd.;

[0059] 1000mL sand core filter device, Tianjin Jinteng Experimental Equipment Co., Ltd.;

[0060] QP-01 small vacuum pump, Shenzhen Liangyi Instrument Company;

[0061] DS-5510 DTH ultrasonic cleaner, Shanghai Aoxi Scientific Instrument Co., Ltd.;

[0062] Agilent 1200series HPLC...

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Abstract

The invention discloses a determination method of the content of endogenous hormones in terminal buds of tomato plants. The method comprises the following steps: 1) preprocessing a tomato terminal bud sample for test; 2) preparing a mobile phase and setting chromatographic conditions; 3) preparing a standard liquid; 4) inspecting specificity; 5) inspecting a linear relation; and 6) determining sample content. The method of the step 1) comprises process of performing vacuum evaporation after extraction and petroleum ether extraction, further regulating pH, carrying out ethyl acetate extraction, and finally performing secondary vacuum evaporation to obtain a to-be-determined sample liquid; and in the step 2), the mobile phase is prepared from methanol and 0.075% glacial acetic acid. The determination method of the content of endogenous hormones in terminal buds of tomatoes by HPLC, provided by the invention, can quickly, accurately and simultaneously determine the content of four plant endogenous hormones, and has the characteristics of fast and efficient determination, high sensitivity and high degree of automation; a recovery of standard addition experiment is carried out, the average recovery is 99.85%, and RSD (Relative Standard Deviation) is less than 1.56%, which meets the hormone determination requirement.

Description

technical field [0001] The invention relates to the technical field of crop endogenous hormones in agronomy, in particular to a method for measuring the content of endogenous hormones in tomato plant terminal buds by high performance liquid chromatography (HPLC). Background technique [0002] Tomato (Lycopersicon esculentum) is one of the main facility crops in the world and in China. It is loved by consumers because of its good taste and rich nutrition. my country's protected tomato cultivation area and output both rank first in the world. At present, most of the scientific research on facility tomato focuses on morphological indicators, photosynthetic characteristics and dry matter distribution. Through the study of molecular mechanism, it is of great scientific significance to deeply understand the mechanism of plant hormones regulating the growth and development of facility tomato and its adaptation to the environment. [0003] Plant hormones (plant hormones, PH) refer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 杨再强袁昌洪邱译萱
Owner 怀远县翰邦科技咨询有限公司
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