CN114705772A - Black tea storage year distinguishing method based on year black tea characteristic compounds - Google Patents
Black tea storage year distinguishing method based on year black tea characteristic compounds Download PDFInfo
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- CN114705772A CN114705772A CN202210299041.XA CN202210299041A CN114705772A CN 114705772 A CN114705772 A CN 114705772A CN 202210299041 A CN202210299041 A CN 202210299041A CN 114705772 A CN114705772 A CN 114705772A
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- black tea
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- theaflavin
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- pyrrolidone
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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Abstract
A black tea storage year distinguishing method based on year black tea characteristic compounds belongs to the technical field of tea year distinguishing. The method comprises the following steps: establishing a linear regression equation of the content of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in the black tea and the year of the black tea, measuring the content of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in a black tea sample of unknown year, and substituting the quantitative result into the linear regression equation so as to obtain the pre-estimated value of the year of the black tea sample. The method has the advantages of simple and quick operation, data quantification, objective, accurate and reliable result and less sample consumption.
Description
Technical Field
The invention belongs to the technical field of tea year discrimination, and particularly relates to a black tea year discrimination method based on characteristic compounds in year black tea.
Background
The black tea belongs to the whole fermentation tea, is one of the traditional six teas in China, is also the tea with the largest consumption in the world, and is popular with consumers due to the unique aroma and mellow taste. The black tea has various pharmacological effects of helping gastrointestinal digestion, preventing gastrointestinal inflammation, losing weight and reducing fat, eliminating edema and the like.
Storage has very important influence on the content and flavor quality of tea leaves, and researches find that the color, aroma and taste of tea soup of the black tea can be greatly changed after the black tea is stored for a long time. Wherein, the color of the tea soup is changed from bright red to dark red, the tea soup gradually shows old fragrance, the bitterness is reduced, and the taste is changed from sweet and mellow to mellow. In recent years, black tea has received a lot of attention from tea workers in the year and is enjoyed by consumers. However, the black tea of different years is difficult to be directly and accurately judged from the aspects of appearance, liquor color, aroma, taste and the like, and the artificial subjective factors are large in the tea sensory evaluation process. In addition, no specific compound index is available for directly distinguishing the annual black tea. Therefore, a simple and fast analysis method for judging the storage years of black tea samples is urgently needed. We found four new components in the storage process of black tea in the previous period, namely N-ethyl-2-pyrrolidone substituted theaflavin, N-ethyl-2-pyrrolidone substituted theaflavin-3-gallate, N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate. The content of the four compounds linearly increases in the storage process of the black tea and has a strong positive correlation with the storage time of the black tea. The four compounds and the storage years are utilized to establish a linear regression equation, and the compounds N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate which greatly contribute to the judgment of the years are screened out. The patent firstly proposes that the content of theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and the content of theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone are adopted to judge the year of black tea, and provides a basis for judging the year of black tea.
Disclosure of Invention
In view of the problems in the prior art, the present invention aims to provide a technical solution for a method for discriminating storage years of black tea based on a compound characteristic of year black tea, which provides effective technical support for discrimination of commercially available year black tea. The method has the advantages of simple and quick operation, data quantification, objective, accurate and reliable result and less sample consumption.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention provides a method for judging the storage years of black tea based on the characteristic compounds of the old black tea, the method is to establish a linear regression equation of the content of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in the black tea and the year of the black tea, by measuring the content of the theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and the theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone in the samples of the black tea in unknown years, and then substituting the quantitative result into the linear regression equation to obtain the estimated value of the year of the black tea sample.
Further, N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -bisThe linear equation of the content of the gallic acid ester and the year of the black tea is as follows: 2.3272x1+0.2441x2-6.4329, where x1Is N-ethyl-2-pyrrolidone substituted theaflavin-3' -gallate, x2Is theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone, and y is black tea year.
Further, the method specifically comprises the following steps:
1) extraction of black tea ingredients: collecting black tea samples with different storage times, grinding into powder, accurately weighing a small amount of black tea powder, fully dissolving and extracting non-volatile chemical components in the black tea powder by using a methanol water solution, centrifuging and filtering by using a filter membrane, and taking supernate;
2) quantitative analysis of the characteristic compounds of the yearly black tea based on liquid chromatography-mass spectrometry, namely the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate;
3) establishing a linear regression equation for judging the black tea year through a series of black tea samples of known years;
4) substituting the quantitative results of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate of the unknown black tea sample into a linear regression equation for judging the black tea year so as to obtain the pre-estimated value of the black tea sample year.
Further, the small amount of black tea powder in the step 1) is specifically 0.1 g.
Further, in the step 1), the proportion of the methanol aqueous solution is 70%, the extraction temperature is 70 ℃, the extraction time is 30 minutes, and the volume of the extraction solvent is 20 milliliters.
Further, the centrifugation condition in the step 1) is 8000 rpm and the centrifugation time is 10 minutes.
Further, the quantitative analysis in the step 2) is based on an external standard method.
Further, the estimated value of the black tea sample year in the step 4) retains 2 decimal parts.
Further, the method for detecting the content of the theaflavin-3 '-gallate substituted by the N-ethyl-2-pyrrolidone and the theaflavin-3, 3' -digallate substituted by the N-ethyl-2-pyrrolidone is an ultra-high performance liquid chromatography-quadrupole orbitrap mass spectrometry method based on metabonomics.
The invention also provides application of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate as characteristic compounds for distinguishing the year of black tea.
The invention judges the year of the black tea by the year black tea characteristic compound, and has the following advantages: 1) the compounds used as the black tea year discrimination indexes are theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone which are found in tea leaves for the first time, and have originality; 2) the artificial subjective factors related to the black tea year discrimination mainly based on the tea sensory evaluation are eliminated, unified quantitative data are provided, the result is objective and reliable, the inspection and analysis can be completed without professional tea leaf evaluation personnel, and the requirements on experimenters are greatly reduced; 3) the experimental operation and data analysis are simple, feasible, quick and efficient.
Drawings
FIG. 1 shows the chemical structures of N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate (A) and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -digallate (B).
FIG. 2 shows an extracted ion chromatogram (A) and a mass spectrum (B) of N-ethyl-2-pyrrolidone-substituted theaflavin-3' -gallate; and (3) extracting an ion current chromatogram (C) and a mass spectrum (D) of the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate.
Detailed Description
The following examples of the present invention are described in detail with reference to the accompanying tables: the present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments.
Example 1:
and (3) identification and judgment of black tea year: the content of N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in the known black tea samples of different years is measured, and then a linear discrimination model is established with the storage years, so that the black tea samples of unknown years are subjected to year identification discrimination.
The method comprises the following specific operation steps:
1) 54 black tea samples of different years were collected, ground into powder and stored in a refrigerator at-20 ℃.
2) Measuring the content of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in the black tea sample:
accurately weighing 0.1g of black tea sample powder into a centrifuge tube, adding 20mL of 70% methanol aqueous solution, fully mixing, leaching in a 70 ℃ water bath for 30 minutes, and shaking once every 5 minutes. Centrifugation is carried out for 10 minutes at 8000 rpm, supernatant is taken to pass through a 0.22 mu m nylon membrane, and then 1.5mL of supernatant is taken to be in a liquid phase sample injection vial for subsequent liquid chromatography-mass spectrometry analysis.
The chromatographic conditions for LC-MS are as follows: t3 column (100 mm. times.2.1 mm, 1.8 μm, Volter, USA) with mobile phase A of 0.1% formic acid-water solution and B of 0.1% formic acid-containing acetonitrile solution; the sample injection amount is 3 mu L; the flow rate was 0.4 mL/min-1(ii) a The column temperature was 40 ℃; the mobile phase linear gradient elution was: 0 min, 2% phase B; 0.5 min, 2% phase B; 10 min, 15% phase B; 18 minutes, 40% phase B; 20 min, 90% phase B; 20.9 min, 90% phase B; 21 min, 2% phase B; 25 min, 2% B phase.
The mass spectrum conditions of the LC-MS are as follows: an electrospray ionization (ESI) source is adopted; the detection mode is a positive ion mode; the capillary voltage is 3500V, and the capillary temperature is 300 ℃; the temperature and flow rate of the auxiliary gas are respectively 350 deg.C and 10L min-1(ii) a The mass spectrum scanning range is mass-to-charge ratio (m/z) 100-1200.
The peak appearance of N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -digallate under the chromatography-mass spectrometry conditions is shown in FIG. 2. The content of the theaflavin-3 '-gallate and the theaflavin-3, 3' -digallate are quantified by an external standard method through standard samples of N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate. The contents of N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -digallate in the 54 black tea samples are shown in Table 1. The content of theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone increases with the age of black tea.
TABLE 1 content of N-ethyl-2-pyrrolidone-substituted theaflavin-3 ' -gallate (TF-3 ' -G-cThea) and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3 ' -digallate (TF-DG-cThea) in black tea year
3) Establishment of linear equation for black tea year discrimination
The contents (mu g/g) of N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate in black tea samples are respectively x1And x2The variables were determined by linear regression analysis using the storage years of the black tea samples as the y variable. Obtaining the linear regression equation y of the year discrimination of the black tea as 2.3272x1+0.2441x2-6.4329，r2＝0.963。
4) Black tea year identification and discrimination
A professional is entrusted to buy 10 representative black tea samples, the year information is kept secret in advance, the operation is carried out according to the steps 1-4 of the instruction book, and the year is judged. The experimental result shows that the year estimated value is well matched with the actual year, the judgment error is 0.05-0.59 years, and the accuracy is high, as shown in table 2.
TABLE 2.10 Black tea samples with the contents of N-ethyl-2-pyrrolidone-substituted theaflavin-3 ' -gallate (TF-3 ' -G-cThea) and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3 ' -digallate (TF-DG-cThea), actual year and predicted year
Claims (10)
1. A method for judging the storage years of black tea based on year black tea characteristic compounds is characterized in that a linear regression equation of the contents of theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone in the black tea and the years of the black tea is established, by measuring the content of the theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and the theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone in the samples of the black tea in unknown years, and then substituting the quantitative result into the linear regression equation to obtain the estimated value of the year of the black tea sample.
2. A method for determining the storage year of black tea based on a characteristic compound of yearly black tea as claimed in claim 1, wherein the linear equation of the content of N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -digallate in black tea with respect to the yearly black tea is: 2.3272x1+0.2441x2-6.4329, where x1Is N-ethyl-2-pyrrolidone substituted theaflavin-3' -gallate, x2Is theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone, and y is black tea year.
3. A method for determining the storage year of black tea based on a compound characteristic of annual black tea as claimed in claim 1, which comprises the steps of:
1) extracting the content of the black tea: collecting black tea samples with different storage times, grinding into powder, accurately weighing a small amount of black tea powder, fully dissolving and extracting non-volatile chemical components in the black tea powder by using a methanol water solution, centrifuging and filtering by using a filter membrane, and taking supernate;
2) quantitative analysis of the characteristic compounds of the yearly black tea based on liquid chromatography-mass spectrometry, namely the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate;
3) establishing a linear regression equation for judging the black tea year through a series of black tea samples of known years;
4) substituting the quantitative results of the N-ethyl-2-pyrrolidone substituted theaflavin-3 '-gallate and the N-ethyl-2-pyrrolidone substituted theaflavin-3, 3' -digallate of the unknown black tea sample into a linear regression equation for judging the black tea year so as to obtain the pre-estimated value of the black tea sample year.
4. A method for determining the storage year of black tea based on the compounds characteristic of yearly black tea according to claim 3, wherein the small amount of black tea powder in the step 1) is 0.1 g.
5. A method for determining the storage year of black tea based on the year black tea specific compound according to claim 3, wherein the ratio of the methanol aqueous solution in the step 1) is 70%, the extraction temperature is 70 ℃, the extraction time is 30 minutes, and the volume of the extraction solvent is 20 ml.
6. A method of determining the storage year of black tea based on a compound characteristic of annual black tea according to claim 3, wherein the centrifugation in the step 1) is carried out at a rotation speed of 8000 rpm for a centrifugation time of 10 minutes.
7. The method for determining the storage year of black tea based on the compounds characteristic of yearly black tea according to claim 3, wherein the quantitative analysis in the step 2) is a quantitative analysis based on an external standard method.
8. A method for determining the storage year of black tea based on the year black tea specific compound according to claim 3, wherein the predicted value of the year of the black tea sample in the step 4) is retained at 2 decimal places.
9. The method for determining the storage period of black tea based on the compounds characteristic of annual black tea according to claim 3, wherein the method for detecting the content of N-ethyl-2-pyrrolidone-substituted theaflavin-3 '-gallate and N-ethyl-2-pyrrolidone-substituted theaflavin-3, 3' -digallate is a metabonomics-based ultra high performance liquid chromatography-quadrupole orbitrap mass spectrometry method.
The application of theaflavin-3 '-gallate substituted by N-ethyl-2-pyrrolidone and theaflavin-3, 3' -digallate substituted by N-ethyl-2-pyrrolidone as characteristic compounds for distinguishing the year of black tea.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2019217269A1 (en) * | 2018-05-08 | 2019-11-14 | Epc Natural Products Co., Ltd. | A tasteful natural sweetener and flavor |
| CN111855880A (en) * | 2019-04-29 | 2020-10-30 | 勐海茶业有限责任公司 | Method for detecting series of puerins 401 compounds in fermented tea, quality control method and method for identifying tea |
| CN112946156A (en) * | 2021-03-10 | 2021-06-11 | 安徽农业大学 | Method for rapidly judging grade of black tea |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019217269A1 (en) * | 2018-05-08 | 2019-11-14 | Epc Natural Products Co., Ltd. | A tasteful natural sweetener and flavor |
| CN111855880A (en) * | 2019-04-29 | 2020-10-30 | 勐海茶业有限责任公司 | Method for detecting series of puerins 401 compounds in fermented tea, quality control method and method for identifying tea |
| CN112946156A (en) * | 2021-03-10 | 2021-06-11 | 安徽农业大学 | Method for rapidly judging grade of black tea |
Non-Patent Citations (2)
| Title |
|---|
| CHEN D 等: "Metabolomics analysis reveals four novel N-ethyl-2-pyrrolidinone-substituted theaflavins as storage-related marker compounds in black tea" * |
| 宁井铭 等: "基于代谢谱分析的祁门红茶加工过程中儿茶素及芳香类物质变化" * |
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