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Method for analyzing chlorophyll and degradation product components thereof in tea leaves

A technology of degradation products and analysis methods, which is applied in analysis materials, material separation, measurement devices, etc., can solve the problems of complex pretreatment, low sensitivity, long time consumption, etc., and achieve simple and rapid pretreatment, high detection coverage, and analysis system. full effect

Pending Publication Date: 2021-08-03
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Spectrophotometry involves complex pretreatments such as acid treatment and saponification, and can only roughly measure chlorophyll components, with low sensitivity
Thin-layer chromatography is time-consuming and poorly separates components
Although liquid chromatography detection can improve the separation between different components, the pretreatment is also complicated and time-consuming
The detection coverage of the above methods for chlorophyll and its degradation components needs to be improved, and it is difficult to simultaneously extract and measure multiple derivative products (water-soluble and fat-soluble) on the chlorophyll degradation pathway, and it is difficult to separate compounds with similar chemical structures and properties. epimer
The limitations of the above technologies are difficult to meet the needs of detailed characterization of chlorophyll components in tea and systematic research on its transformation process

Method used

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  • Method for analyzing chlorophyll and degradation product components thereof in tea leaves
  • Method for analyzing chlorophyll and degradation product components thereof in tea leaves
  • Method for analyzing chlorophyll and degradation product components thereof in tea leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1) Extraction of chlorophyll and its degradation products in the tea to be tested

[0041] ① After the tea sample to be tested is ground, pass through a 30-mesh sieve and set aside;

[0042]②Weigh about 20.0mg±0.1mg of tea powder into a centrifuge tube, add 300μL methanol solution containing internal standard compound, and vortex for 30s. The internal standard compound is lysophosphatidylcholine LPC19:0, the concentration is 2.00 μg / mL; phosphatidylcholine PC38:0, the concentration is 1.33 μg / mL;

[0043] ③ Add 1mL MTBE, vortex for 30min;

[0044] ④Add 300μL ultrapure water and vortex for 30s;

[0045] ⑤ After standing for 10 minutes, centrifuge at high speed (10000rpm*10min) to make layers;

[0046] ⑥ Take 600 μL of the upper layer and 250 μL of the lower layer, transfer to a new centrifuge tube, concentrate and dry by low-temperature centrifugation, and set aside.

[0047] 2) LC-MS analysis of chlorophyll and its degradation products in the tea to be tested

[00...

Embodiment 2

[0057] Example 2: Chlorophyll degradation pathway analysis in green tea

[0058] (1) Get a green tea sample and analyze it according to the method in Example 1. Three copies of each tea sample were taken for testing as technical replicates.

[0059] (2) Analysis of chlorophyll degradation pathways in green tea: the results showed that all 19 chlorophylls and their degradation products involved in the present invention could be detected in green tea samples, covering enzymatic and non-enzymatic degradation products of different properties. Path analysis shows that ( figure 1 ), green tea processing involves the following reactions: ① Isomerization: Chlorophyll a and b are converted into their respective isomers a' and b'. ②Enzyme-catalyzed explantation reaction: under the action of chlorophyllase, a hydrolysis reaction occurs to generate water-soluble explantation chlorophyll. ③ Demagnesium degradation reaction: Under acidic or heating conditions, the Mg on the porphyrin rin...

Embodiment 3

[0060] Example 3: Comparative analysis of chlorophyll and its degradation products in green tea under different foot fire methods

[0061] (1) Making green tea: spread naturally (indoor temperature 25-28°C, relative humidity 70%-80%, wait until the moisture content drops to about 70%) → drum-type finishing (leaf throwing amount 150kg / h, 260°C, 75s )→cooling (1h)→kneading (30kg leaves, rotating speed 50-52r / min, lightly kneading for 30min)→hair drying (110°C, 20min, until the moisture content drops to about 20%). Then use five different foot fire methods (far-infrared foot fire, microwave foot fire, aroma machine foot fire, strip machine foot fire and hexagonal hui pot foot fire) to carry out foot fire drying to obtain green tea.

[0062] (2) The green tea made by different foot fire methods is analyzed according to the method in Example 1. Three copies of each tea sample were taken for testing as technical replicates.

[0063] (3) Orthogonal partial least squares discriminan...

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Abstract

The invention discloses a method for analyzing chlorophyll and degradation product components thereof in tea and belongs to the field of food or plant quality component analysis and detection. The method comprises the following steps of: extracting chlorophyll and degradation products thereof in tea leaves to be detected; performing LC-MS analysis on the chlorophyll and degradation products thereof in the tea leaves to be detected; and carrying out relative quantitative analysis on the chlorophyll and degradation products thereof in the tea leaves to be detected. According to the method, sample pretreatment is simple and rapid, and chlorophyll with different solubilities and degradation product components (water-soluble and fat-soluble components) thereof can be extracted at the same time; the analysis speed is high, and the LC-MS analysis time of each sample is only 20 minutes; and the detection coverage degree of the chlorophyll and degradation product components thereof in the tea leaves is high, and the comprehensive coverage of 19 components in enzyme-mediated degradation and non-enzyme thermal degradation means in a chlorophyll degradation path can be realized. In short, the method is simple and rapid, and is more systematic and comprehensive in analysis.

Description

technical field [0001] The invention belongs to the field of analysis and detection of food or plant quality components, and in particular relates to an analysis method of chlorophyll and its degradation product components in tea. Background technique [0002] Tea color is one of the important factors of tea quality attributes of "color, aroma, taste and shape". It is the most direct visual perception of consumers. The beautiful and smooth color is an important basis for consumers to choose. Tea color includes three aspects: dry tea, soup color and leaf bottom. Taking green tea as an example, green tea is loved by consumers because of its "three green" quality characteristics, including tea soup green, dry tea green, and leaf bottom green. The coloring substances of tea include chlorophyll and its degradation derivatives, carotenoids, flavonoids, tea pigments (theaflavins, tea red, theabrownins), etc.; because of their different solubility, they can also be divided into wat...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/96
CPCG01N30/02G01N30/06G01N30/96G01N2030/065G01N2030/027
Inventor 李佳吴仕敏江用文袁海波邓余良滑金杰杨艳芹王近近朱佳依沈帅
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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