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Method for determining four hormones in rubber tree through high-efficiency liquid chromatography

A high-performance liquid chromatography, a rubber tree technology is applied in the field of high-performance liquid chromatography for the determination of four hormones in rubber trees, which can solve problems such as poor elution effect, error in determination results, dragging of peaks, etc. The effect of high accuracy and precision, and short extraction process time

Inactive Publication Date: 2015-04-15
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the chromatographic analysis process, only two organic solvents, methanol and acetonitrile, are prone to peak dragging, and the elution effect is not good, resulting in errors in the measurement results.

Method used

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  • Method for determining four hormones in rubber tree through high-efficiency liquid chromatography
  • Method for determining four hormones in rubber tree through high-efficiency liquid chromatography
  • Method for determining four hormones in rubber tree through high-efficiency liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, the content of four kinds of hormones of auxin, zeatin, gibberellin and abscisic acid in Hevea brasiliensis leaves determined by high performance liquid chromatography

[0059] 1. Collection and pretreatment of Hevea brasiliensis leaves

[0060] The leaves of Hevea brasiliensis were collected from Reyan 7-33-97, a clone of Hevea brasiliensis planted in the experimental field of the Chinese Academy of Tropical Agricultural Sciences. The sample pretreatment was carried out as described above to obtain the samples to be tested.

[0061] 2. High performance liquid chromatography to detect the content of four hormones in the sample to be tested

[0062] Chromatographic conditions: the chromatographic column is CNWSIL C18 liquid chromatographic column (4.6×150mm5μm) produced by American Waters Company, the column temperature is 30°C; the mobile phase is methanol, acetonitrile, phosphate buffer (volume ratio is 15:15:70, pH3 .5) Isocratic elution; the flow rate...

Embodiment 2

[0078] Embodiment 2, the content of four kinds of hormones of auxin, zeatin, gibberellin and abscisic acid in the rubber tree branch of high performance liquid chromatography

[0079] 1. Collection and pretreatment of rubber tree branches

[0080] Brasiliensis branches were collected from the Hevea brasiliensis clone Reyan 7-33-97 planted in the experimental field of the Chinese Academy of Tropical Agricultural Sciences. The sample pretreatment was carried out as described above to obtain the samples to be tested.

[0081] 2. High performance liquid chromatography to detect the content of four hormones in the sample to be tested

[0082] The operation method is the same as step 2 in Example 1.

[0083] Chromatograms such as image 3As shown, the hormone content in the shoots of Hevea brasiliensis clone Reyan 7-33-97 was further determined according to the standard curve as follows: the content of zeatin (ZT) was 1581.6 μg / g fresh weight; gibberellin (GA3 ) content was 5.06...

Embodiment 3

[0084] Embodiment 3, the content of four kinds of hormones of auxin, zeatin, gibberellin and abscisic acid in the bark of Hevea brasiliensis determined by high performance liquid chromatography

[0085] 1. Collection and pretreatment of the bark of Hevea brasiliensis

[0086] The bark of Hevea brasiliensis was collected from Reyan 7-33-97, a clone of Hevea brasiliensis planted in the experimental field of Chinese Academy of Tropical Agricultural Sciences. The sample pretreatment was carried out as described above to obtain the samples to be tested.

[0087] 2. High performance liquid chromatography to detect the content of four hormones in the sample to be tested

[0088] The operation method is the same as step 2 in Example 1.

[0089] Chromatograms such as Figure 4 As shown, the hormone content in the bark of Hevea brasiliensis clone Reyan 7-33-97 was further determined according to the standard curve as follows: the content of zeatin (ZT) was 102.9 μg / g fresh weight; gi...

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Abstract

The invention discloses a method for determining four hormones in a rubber tree through high-efficiency liquid chromatography. The method provided by the invention comprises the following steps of: (1) sample pretreatment: to be specific, grinding rubber tree tissues with liquid nitrogen, then placing in leaching liquor for extraction, centrifuging, separating a centrifuged liquid supernatant to obtain an aqueous phase, enabling the aqueous phase to pass through a chromatographic column, and eluting with methyl alcohol to obtain a sample to be tested; and (2) high-efficiency liquid chromatography detection: adopting an inverse-phase C18 chromatographic column, taking a mixed liquid of the methyl alcohol, acetonitrile and phosphate buffer as a mobile phase and 254nm as testing wavelength, and detecting auxin, zeatin, activol and abscisic acid in the sample to be tested obtained in the step (1). The method provided by the invention has the advantages that the four hormones can be detected, the extraction flow time is short, the determination accuracy rate is high, and the determination and analysis efficiency is improved. The method is a good method for researching the high-yield fast-growing mechanism of the Brazil rubber tree and the plant hormones in the cambium differentiation process.

Description

technical field [0001] The present invention relates to a method for measuring four hormones in rubber tree by high performance liquid chromatography, in particular to a method for simultaneously measuring auxin, zeatin, gibberellin and abscisic acid in rubber tree leaves, branches or bark by using high performance liquid chromatography The four hormonal approach. Background technique [0002] The natural rubber industry has long focused on collecting latex as industrial raw materials, supplemented by the development and utilization of other biomass such as wood and seeds. Endogenous hormones auxin (IAA), gibberellin (GA 3 ), the content, activity and signaling pathway of abscisic acid (ABA) and zeatin (ZT) regulate latex yield and wood growth and development. However, the above-mentioned four endogenous hormones of rubber tree are extremely low in content and unstable in nature, and there are interferences of various components in rubber tree leaves and bark, which makes ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 王立丰
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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