HPLC-DAD segmented detection method for simultaneously determining 14 polyphenol compounds in tobacco

A polyphenolic compound and detection method technology, applied in the field of HPLC-DAD segmented detection, can solve the problems of shortening the service life of a chromatographic column or an instrument, failing to detect mass fraction of polyphenolic compounds, etc., and achieves high sensitivity and peak shape. Good, reduce the effect of interference

Active Publication Date: 2020-09-29
CHINA TOBACCO GUANGDONG IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome that the existing tobacco polyphenol detection technology can only detect about 10 kinds of polyphenol compounds at the same time, and other polyphenol compounds with lower mass fractions cannot be detected, and the mobile phase used will shorten the chromatogram. Defects and deficiencies such as the service life of the column or instrument provide a HPLC-DAD segmented detection method with high sensitivity, precision and accuracy that can simultaneously determine 14 polyphenolic compounds in tobacco

Method used

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  • HPLC-DAD segmented detection method for simultaneously determining 14 polyphenol compounds in tobacco
  • HPLC-DAD segmented detection method for simultaneously determining 14 polyphenol compounds in tobacco
  • HPLC-DAD segmented detection method for simultaneously determining 14 polyphenol compounds in tobacco

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The optimization of embodiment 1 chromatographic conditions

[0062] 1.1 Selection of mobile phase

[0063] Due to the active chemical properties of polyphenolic compounds, phenolic hydroxyl groups are easily oxidized and ionized. In the environment where methanol-water is used as the mobile phase, polyphenols will be ionized as a weak electrolyte, and there is a dual retention mechanism on the surface of the stationary phase. poor effect. It has also been confirmed in practice that neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid have severe tailing of chromatographic peaks without adding appropriate additives to the mobile phase.

[0064] In the case of other chromatographic conditions being the same, mobile phase system I (mobile phase A: 93% water + 4.5% methanol + 2.5% acetic acid; mobile phase B: 10% water + 88% methanol + 2% acetic acid) and mobile phase Phase system II (mobile phase A: 0.05mol / L potassium dihydrogen phosphate aqueous solution...

Embodiment 2

[0103] The selection of embodiment 2 extraction conditions

[0104] 2.1 Selection of solvent ratio and extraction time

[0105] After pulverizing 0.02g of tobacco (accurate to 0.0001g), put it in a 10mL stoppered glass vial, add 5mL methanol solution with a concentration of 30-100vol% and ultrasonically extract it for 10-50min, take the supernatant and filter it through a 0.22μm water phase Membrane, the tobacco sample to be tested was obtained, and the optimal chromatographic conditions in 1.4 were used for detection.

[0106] See results Figure 8 , the picture shows 8 kinds of polyphenols with higher content in tobacco, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, scopoletin, caffeic acid, scopoletin, rutin and kaempferol-3-O-rutinoside The extraction effect of the compound under different methanol / water ratios, the other 6 polyphenols are low in content, which is close to a straight line in the figure, so it is not shown in the figure. Depend on Figu...

Embodiment 3

[0110] Embodiment 3 methodological investigation

[0111] 3.1 Determination of linear range, detection limit and quantification limit

[0112] Use the optimal chromatographic conditions in 1.4 to measure the series of standard solutions, and perform linear regression analysis on the chromatographic peak area and concentration of the target compound to obtain the regression equation and its related parameters. Three times of the deviation is the detection limit of the method, and 10 times of the standard deviation of the obtained measurement results is the quantitative limit of the method. The results are shown in Table 2.

[0113] Table 2 linear relationship, detection limit, limit of quantitation determination result

[0114]

[0115] Note: The correlation coefficient R of the regression equation 2 =0.99976~0.99997.

[0116] It can be seen from Table 2 that within the test range, the correlation coefficient between the response of the target substance and its concentrat...

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Abstract

The invention belongs to the technical field of tobacco chemical component analysis, and particularly relates to an HPLC-DAD segmented detection method for simultaneously determining 14 polyphenol compounds in tobacco. According to the method, a potassium dihydrogen phosphate aqueous solution and methanol are used as mobile phases; gradient elution is carried out according to a specific proportion; and 14 polyphenol compounds in the tobacco can be simultaneously determined, so that the segmented wavelength is further limited, the signal intensity of the polyphenol compounds can be remarkably improved, the sensitivity is high, the peak pattern of a target peak is good, the separation degree is high, the determination of low-content polyphenol compounds in the tobacco is facilitated, and theinterference of other impurities is reduced; and it is proved by methodological investigation that the method provided by the invention has high precision, good reproducibility and high accuracy, and can be used in tobacco quality control and sensory characteristic analysis.

Description

technical field [0001] The invention belongs to the technical field of tobacco chemical component analysis. More specifically, it relates to an HPLC-DAD segmented detection method for the simultaneous determination of 14 polyphenolic compounds in tobacco. Background technique [0002] Polyphenols are a class of important compounds in tobacco, and their content in tobacco can be as high as 5%. They play an important role in the growth and development of tobacco, the characteristics of modulation, the color of tobacco leaves, the taste of smoke, and the physiological strength of smoke. For example, chlorogenic acid is the dicondensate of caffeic acid and quinic acid, which itself has a weak green fragrance. It forms polyquinones under the action of polyphenol oxidase, and undergoes a non-enzymatic browning reaction with amino acids to generate pyrazine and pyridine. , pyrroles, etc. These substances have a nutty, toasted or bready aroma, and are characteristic aroma substance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/74G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/74G01N30/8679
Inventor 赖燕华欧阳璐斯王予陶红潘晓薇黄翼飞周瑢陈翠玲汪军霞
Owner CHINA TOBACCO GUANGDONG IND
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