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Method for measuring content of lignin monomers in pear peel

A determination method and monomer content technology, applied in the biological field, can solve the problems of inability to separate, measure, and lack of lignin monomers, and achieve the effects of reducing sample browning, early peaking time, and accurate quantification.

Inactive Publication Date: 2015-04-22
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] At present, the determination methods for lignin content mainly include acid hydrolysis method, mercaptoacetic acid method and acetyl bromide method, etc., but these methods are all aimed at determining the total lignin content, and cannot separate and measure lignin monomers.
And for lignin monomer, only the mensuration of lignin monomer content in the root of traditional Chinese medicine Salvia miltiorrhiza is found in the report at present, in aspect the lignin monomer content determination in the pear peel, lacks a kind of effective, simple and accurate assay method

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  • Method for measuring content of lignin monomers in pear peel
  • Method for measuring content of lignin monomers in pear peel

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Embodiment

[0037] 1 Instruments and samples

[0038] 1.1 Instrument: Agilent 7890A GC system connected in series with 5975C inert XL MSD. Chromatographic column: Agilent HP-5, 30m*0.25mm*0.25μm.

[0039] 1.2 Sample: the peel of 'Dangshansu pear' and its brown-skinned bud variety 'Xussu'.

[0040] 2 Methods and results

[0041] 2.1 Sample treatment: Take pear peels of about 1 mm thickness from the above samples, grind them with liquid nitrogen, extract them with acetone and methanol for 24 hours, and then extract them with hot water for 30 minutes. Dry the samples at 45°C for later use. Weigh 50 mg of the dried sample, add 1 mL of freshly prepared reaction solution (final concentration of 2.5% boron trifluoride ether and 10% ethanethiol dissolved in dioxane), and place in a constant temperature oven at 100°C for reaction 4h, then placed at -20°C to stop the reaction. Centrifuge at 12000 rpm for 10 min, take 300 μL of the supernatant, add 0.2 mL of n-docosane (dissolved in dichlorometh...

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Abstract

The invention provides a method for measuring the content of lignin monomers in pear peel. The method comprises the following steps: grinding a sample by using liquid nitrogen, performing organic phase and hot water extraction, and drying; extracting by adopting boron trifluoride and ethanethiol, adding internal standard n-docosane, drying, deriving by using pyrimidine and N,O-bis(trimethylsilyl)acetamide after drying, and analyzing a gas chromatograph-mass spectrometer; performing qualitative analysis according to the molecular ion peak, wherein the mass-to-charge ratio (m / z) 269 refers to the G lignin monomer, and the m / z 299 refers to the S lignin monomer; and quantifying according to the concentration of n-docosane, the peak area and the gas phase response factor, and calculating to obtain the content of the two lignin monomers in the sample. The method for measuring the content of the lignin monomers in the pear peel, provided by the invention, is easy to operate, a basis can be provided for researching the influence of the lignin monomers in the peel on the peel color, and a theoretical basis is laid for further controlling the peel color.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for determining the content of lignin monomers in pear peel. Background technique [0002] Lignin is divided into syringyl lignin (S-lignin) polymerized from syringyl propane structural monomer, guaiacyl lignin (G- Lignin) and p-hydroxyphenyl lignin (H-lignin) polymerized from p-hydroxybenzylpropane structural monomers. In most angiosperms including pear, only S-lignin and G-lignin exist in the species. [0003] The color of pear peel is an important indicator of the economic value of the fruit. Studies have shown that the brown formation of pear peel is closely related to lignin synthesis. With the reduction of the activity of related enzymes PPO and POD, the amount of lignin synthesis decreases significantly, and in the lignin biosynthesis pathway, S- and G-lignin alone Body formation is regulated by different genes. Therefore, in order to study the formation mechanism...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 衡伟朱立武王孟东叶振风贾兵刘莉刘普
Owner ANHUI AGRICULTURAL UNIVERSITY