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Method measuring C6-C3 type phenolic acids compound content in tobacco

A technology of C6-C3 and phenolic acids, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of unreported and undiscovered patents of C6-C3 phenolic acid compound content methods, and achieve easy operation, Determination of rapid and reproducible effects

Inactive Publication Date: 2013-06-05
GUIZHOU TOBACCO SCI INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] After searching, no patent on the method for determining the content of C6-C3 phenolic acid compounds in tobacco was found in the Chinese patent database, and there is no relevant report in the existing technical literature

Method used

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  • Method measuring C6-C3 type phenolic acids compound content in tobacco

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 Measurement method embodiment

[0025] First prepare the standard solution: Accurately weigh 10 mg of the sinapinic acid standard product, dilute to 10 mL with acetonitrile, then absorb 1 mL and dilute to 10 mL with acetonitrile to make a standard solution for later use;

[0026] Then prepare test sample solution, including:

[0027] (1) Preparation of fresh tobacco leaf samples: take liquid nitrogen samples, then put the fresh tobacco samples into a freeze dryer to freeze-dry until constant weight, that is, no moisture, and then grind them into powder for later use;

[0028] (2) Prepare the cured tobacco leaf samples: the cured tobacco leaves are crushed and passed through a 40-mesh sieve for later use;

[0029] (3) Extract the substance to be tested: take 0.6g of tobacco leaf powder, weigh it accurately, add 30mL of 4mol / L NaOH and shake on a shaker for 4h, add 4mol / L HCl to the supernatant after centrifugation to adjust the pH to 2, and Add NaCl to sat...

Embodiment 2

[0034] Example 2 Linear relationship investigation and selection of detection wavelength and mobile phase

[0035] Take 0.5, 1, 2, 3, 5, 10, 20, 30, and 50 μL of the above-mentioned standard solution respectively, measure according to the chromatographic conditions in Example 1, and record the chromatogram and peak area. Taking the concentration of each standard substance as the abscissa and the peak area as the ordinate to calculate, obtain the regression equation of caffeic acid, p-coumaric acid, ferulic acid and sinapinic acid, listed in the following table:

[0036] substance standard curve line R 2 Linear range detection limit caffeic acid y=8.16×10 3 x-1.82×10 3 0.9998 1.113~56.236 0.314 p-coumaric acid y=1.48×10 4 x-1.45×10 3 0.9993 1.047~52.332 0.572 ferulic acid y=7.55×10 3 x-5.99×10 2 0.9995 1.110~56.620 0.664 sinapinic acid y=5.69×10 3 x-7.18×10 2 0.9992 1.067~53.881 0.955

[0037] Precisi...

Embodiment 3

[0042] Embodiment 3 extraction method selection

[0043] 1. Extraction of free C6-C3 phenolic acid compounds

[0044] In this test, methanol, 80% methanol, and 70% methanol were used as extraction solutions, and ultrasonic waves were used for 30 minutes, 1 hour, and 2 hours to extract free C6-C3 phenolic acid compounds in tobacco, but it was found that tobacco was not detected within the detection limit. Medium C6-C3 free state phenolic acid compounds.

[0045] 2. Extraction of bound C6-C3 phenolic acid compounds

[0046] In this test, 1mol / L, 2mol / L and 4mol / L NaOH were used as the extracting solution, and the stirring extraction and shaking table extraction were used for 1h, 2h, 4h, 8h and 16h respectively, and the extractant used ether and 1:1 ether respectively. : Ethyl acetate and ethyl acetate are used to extract the bound C6-C3 phenolic acid compounds in tobacco. It is found that the extraction effect is better and the extraction time is shorter with 4mol / L NaOH an...

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Abstract

The invention discloses a method measuring C6-C3 type phenolic acids compound content in tobacco. The method relates the following steps: firstly preparing standard substance solution; then preparing sample solution, preparing fresh tobacco leaf samples, preparing samples of baked tobacco leaves, and drawing to-be-measured substance; and conducting content measurement finally, respectively taking the standard substance solution of different concentration to add the standard substance solution to a super-efficient liquid chromatograph, calculating standard curves according to peak areas in chromatograms, and respectively calculating content of caffeic acid, p-cumaric acid, ferulic acid and sinapic acid. The method achieves measurement of the caffeic acid, the p-cumaric acid, the ferulic acid and the sinapic acid, resolves the problem of tobacco quality identification, has the advantages of being simple and convenient to operate, high in sensitivity, strong in accuracy, good in repeatability, high in measuring speed, and the like, can be used for measurement of the caffeic acid, the p-cumaric acid, the ferulic acid and the sinapic acid of the fresh tobacco leaves and the baked tobacco leaves.

Description

technical field [0001] The invention relates to planting and cultivation of tobacco, in particular to a method for measuring the content of C6-C3 type phenolic acid compounds in tobacco. Background technique [0002] The main skeleton types of phenolic acid compounds found in nature are the following two types: (1) C6- [0003] C1 type, the basic skeleton is benzoic acid, such as protocatechuic acid, vanillic acid, syringic acid and gallic acid, etc.; (2) C6-C3 type, mainly cinnamic acid derivatives, such as caffeic acid, ferulic acid, mustard acid, p-coumaric acid, etc. The C6-C3 phenolic acid compounds in tobacco are a class of metabolite components containing phenolic hydroxyl and carboxyl groups, intermediate products of the phenylpropane metabolic pathway in tobacco, and metabolites of the lignin synthesis pathway, mostly in bound form. Generally linked by ester bond, glycosidic bond, ether bond and other compounds, it is closely related to physiological activities su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/36
Inventor 赵会纳丁福章雷波任竹张捷潘文杰罗宝昌
Owner GUIZHOU TOBACCO SCI INST
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