CN107843669B - Method for rapidly determining aroma components of tea - Google Patents
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- CN107843669B CN107843669B CN201711278162.1A CN201711278162A CN107843669B CN 107843669 B CN107843669 B CN 107843669B CN 201711278162 A CN201711278162 A CN 201711278162A CN 107843669 B CN107843669 B CN 107843669B
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Abstract
The invention discloses a method for rapidly measuring tea aroma components, which comprises the following steps: (1) putting the weighing cups containing the tea samples to be tested into a micro-cell thermal extraction instrument in parallel, inserting adsorption tubes in parallel above the corresponding micro-cells, and continuously introducing high-purity carrier gas with a constant flow rate under a certain temperature condition to extract aroma components of the tea; (2) and (3) directly carrying out GC-MS detection after the sample adsorption tube subjected to the micro-cell thermal extraction treatment is thermally desorbed. According to the invention, the micro-cell thermal extraction technology is combined with the thermal desorption and gas chromatography-mass spectrometry technology to extract and analyze the tea aroma components for the first time, the pretreatment of the sample is simple and rapid, the complex pretreatment process is avoided, an organic solvent is not required, the environment is protected, the safety is high, and the sample dosage is small. The tea aroma component detection method established by the invention overcomes the defects of the prior art, and provides a simple, efficient, rapid and green method for extraction, separation and analysis of tea aroma components.
Description
Technical Field
The invention belongs to the field of chemical analysis, relates to an analysis method of tea aroma components, and particularly relates to a method for rapidly analyzing tea aroma components by combining a micro-cell thermal extraction technology with a thermal desorption and gas chromatography-mass spectrometry technology.
Background
The quality of tea generally refers to the color, aroma, taste, shape and bottom of tea. As for tea leaves as a beverage, the aroma and taste of tea soup are the core of tea leaf quality in terms of drinking needs, wherein the aroma is an important factor for determining the quality of tea leaves, and is also the most important factor for capturing and culturing customer loyalty and is an important factor for determining the price of tea leaves. Therefore, the research on the aroma components of the tea has great significance, the analysis and the measurement of the aroma are always an important subject in the field of tea scientific research, and a great deal of research is carried out on the aroma components by tea workers at home and abroad. Heretofore, over 700 kinds of aroma substances including over ten kinds of compounds such as alcohols, aldehydes, ketones, esters, acids, nitrogen heterocycles and the like have been separated from various tea leaves. The first step of the research on the aroma is to extract and separate the aroma, and the first step is directly related to the qualitative and quantitative analysis result of the aroma. The tea has low content of aroma substances, complex components, easy volatilization and instability, and is easily subjected to complex chemical reactions such as oxidation, condensation, polymerization, group transfer and the like in the extraction process due to the influence of external conditions.
Typical methods for extracting aroma include solid-liquid Extraction (such as Soxhlet Extraction, and liquid-liquid Extraction (such as Extraction with a Deanstar apparatus)), Supercritical Fluid Extraction (SFE Method), Simultaneous Distillation Extraction (SDE Method), Vacuum Distillation (VDE Method), and Headspace adsorption (HAS Method). The existing methods for extracting aroma substances have some problems, such as large solvent consumption, time consumption, complicated operation steps or low extraction rate, the extract cannot well reflect the aroma characteristics of tea samples, and the number of aroma components, the total amount of aroma and the relative content of each component obtained by different extraction methods have great difference. Therefore, it is very important to develop a simple, fast, efficient, green and pollution-free extraction technology.
Disclosure of Invention
The invention provides a rapid and effective detection method, which can simultaneously detect more than 80 aroma components in tea.
A method for rapidly measuring tea aroma components comprises the following steps:
(1) putting the weighing cups containing the tea samples to be tested into a micro-cell thermal extraction instrument in parallel, inserting adsorption tubes in parallel above the corresponding micro-cells, and continuously introducing high-purity carrier gas with a constant flow rate under a certain temperature condition to extract aroma components of the tea;
(2) and (3) directly carrying out GC-MS detection after the sample adsorption tube subjected to the micro-cell thermal extraction treatment is thermally desorbed. The micro-cell thermal extraction technology (mu-CTE) is simple and convenient, has high efficiency, easy operation and strong matching property, is suitable for measuring various spices and odor substances, can simultaneously collect 6 samples, has short collection time, can heat and regulate the temperature range from room temperature to 250 ℃, and adopts a unique constant flow control technology to ensure that air or other carrier gas passes through each micro-cell under constant flow.
The invention adopts a micro-tank thermal extraction technology to extract the aroma components of the tea, is simple, convenient, rapid, green and solvent-free, has high enrichment efficiency and small matrix interference, overcomes the defects of large solvent consumption, time consumption, complicated operation steps or low extraction rate and the like of the traditional extraction method, and can well reflect the original aroma characteristics of the tea sample by the extracted volatile matters.
Preferably, the temperature of the micro-pool thermal extraction treatment is 40-90 ℃ and the time is 5-20 min.
Further preferably, the temperature of the micro-pool thermal extraction treatment is 75-85 ℃ and the time is 14-16 min.
Preferably, the sampling amount of the tea sample to be detected is 0.5-1.5 g; more preferably, the sampling amount of the tea sample to be detected is 0.8-1.2 g.
Most preferably, the sampling amount of the tea sample to be tested is 1g, the temperature of the micro-pool thermal extraction treatment is 80 ℃, and the time is 15 min.
Preferably, 6 weighing cups filled with tea samples are placed in 6 micro-pools in parallel, adsorption tubes are inserted into the tops of the corresponding micro-pools side by side, the pressure of each adsorption tube is adjusted to be 35-45 psi, the adsorption tubes are pulled down after being blown for 5-20 min by using high-purity nitrogen as carrier gas, and the adsorption tubes are sealed by brass caps so as to be used for thermal desorption analysis.
Further preferably, the adsorption tube is subjected to an aging treatment before use: introducing high-purity N2Placing the adsorption tubes into an aging instrument, keeping the flow rate of each adsorption tube at 50-100mL/min, adjusting the pressure to 15-30psi, and heating to 300-350 ℃ for 20-40 min.
Further preferably, high purity N is introduced2The adsorption tubes are placed in an aging instrument, the flow rate of each adsorption tube is kept at 50-100mL/min (the regulated pressure is about 15-30psi), and the temperature is raised to 335 ℃ and kept for 30 min.
The microcell thermal extraction is influenced by various factors such as sample amount, extraction temperature, extraction time, carrier gas flow and the like, and in order to realize the optimization of extraction efficiency, the invention optimizes the parameters to ensure that the aroma components in the tea leaves are more effectively analyzed.
Preferably, the thermal desorption conditions are: the Pre-purge time is 1min, the flow rate is 20mL/min, and the split ratio is 6: 1; the desorption temperature is 275 ℃, the desorption time is 5min, the desorption flow is 50mL/min, and no flow distribution is realized; the cold trap analysis temperature is 290 ℃, the flow rate is 20mL/min, and the split ratio is 6: 1.
Preferably, the GC-MS conditions are:
chromatographic conditions are as follows: capillary chromatography column HP-5ms Ultra Inert,325 ℃ (30m × 250 μm × 0.25 μm); temperature programming: setting the initial column incubator at 40 deg.C, maintaining for 0min, heating to 88 deg.C at 3 deg.C/min, maintaining for 0min, then heating to 290 deg.C at 10 deg.C/min, and maintaining for 10 min; carrier gas: helium (99.999% pure); flow rate of carrier gas: 1.3 mL/min; and (3) sample introduction mode: no split-flow sample introduction.
Mass spectrum conditions: an ion source: electron impact ion source (EI source); electron energy: 70 eV; transmission line temperature: 280 ℃; ion source temperature: 230 ℃; temperature of the quadrupole rods: 150 ℃; the mass scanning range is 33-550 amu, the acquisition frequency is 50Hz, and the solvent delay is 2 min.
Preferably, the GC-MS detection result is calculated according to a peak area normalization method, and the relative percentage content of each component is calculated.
Qualitative analysis is carried out through an automatic mass spectrum deconvolution qualitative system, the peak width of a deconvolution parameter is set to be 20, the requirements of resolution, sensitivity and chromatographic peak shape are set to be medium, the minimum value of a matching factor is set to be 80, a spectrogram is searched for NIST14, and meanwhile, auxiliary qualitative analysis is combined with literature. The quantitative analysis is calculated by a chemical workstation data processing system according to a peak area normalization method to obtain the relative percentage content of each component.
Compared with the prior art, the method has the following beneficial effects:
(1) according to the invention, the micro-cell thermal extraction technology is combined with the thermal desorption and gas chromatography-mass spectrometry technology to extract and analyze the tea aroma components for the first time, the pretreatment of the sample is simple and rapid, the complex pretreatment process is avoided, an organic solvent is not required, the environment is protected, the safety is high, and the sample dosage is small.
(2) The tea aroma component detection method established by the invention continuously sweeps out tea aroma components by using flowing nitrogen, has the advantages of high enrichment efficiency and small interference of a matrix, can well reflect the aroma characteristics of a tea sample by using the extract, is particularly suitable for extracting some trace components in tea, and is easy to realize online detection.
(3) The tea aroma component detection method established by the invention can be used for simultaneously carrying out hot extraction on 6 tea samples, so that the unification of experimental conditions is ensured, and experimental errors caused by the change of external factors are avoided.
(4) The tea aroma component detection method established by the invention overcomes the defects of the prior art, and provides a green, rapid, simple and efficient method for extraction, separation and analysis of tea aroma components.
Drawings
FIG. 1 is a total ion flow diagram of aroma components of West lake Longjing tea
FIG. 2 is a total ion flow diagram of aroma components of Mengding camellia
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the scope of the present invention.
Example 1
1. Instruments and reagents
Microcell thermal extractor mu-CTE (Markes, England), aging apparatus TC-20 (Markes, England), thermal desorption apparatus TD-100 (Markes, England), 7890A-5975C gas chromatography-mass spectrometry (Agilent technologies, USA), Tenax-TA pipe (Markes, England), tea (provided by the institute of tea, Chinese academy of agricultural sciences), weighing cup (40 × 25mm, Bodeb Biotech, Beijing), electronic balance (SQP type, Sadous scientific instruments, Ltd.)
2. Extraction of volatile components by micro-cell thermal extraction technology
The method comprises the following specific steps:
(a) aging of Tenax-TA tubes
Before the experiment, an aging instrument is needed to age the Tenax-TA tube, and the method comprises the following specific steps: is introduced intoHigh purity N2The adsorption tubes are placed in an aging instrument, the flow rate of each adsorption tube is kept at 50-100mL/min (the regulated pressure is about 15-30psi), and the temperature is raised to 335 ℃ and kept for 30 min.
(b) Micro-cell thermal extraction
Weighing 1g of tea sample, putting the tea sample into a weighing cup, putting the weighing cup into a micro-cell, setting the extraction temperature to 80 ℃, inserting an adsorption tube into the micro-cell, adjusting the pressure of each tube to be about 40psi, purging for 15min by using high-purity nitrogen as carrier gas, pulling out the adsorption tube, and sealing by using a brass cap so as to be used for thermal desorption analysis. The experimental process optimizes the hot extraction conditions of the micro-cell, mainly considers the factors of sample amount (0.5, 1.0 and 1.5g), extraction time (5, 10 and 15min) and extraction temperature (40, 60 and 80 ℃), adopts three-factor three-level orthogonal experimental design, takes the total peak area as an evaluation index, optimizes and selects the factors shown in table 1, and finally optimizes the experimental conditions that 1g of tea sample is extracted for 15min at 80 ℃.
TABLE 1 results of optimization of various factors of the microcell thermal extraction technology
3. Thermal desorption analysis conditions:
the Pre-purge time is 1min, the flow rate is 20mL/min, and the split ratio is 6: 1; the desorption temperature is 275 ℃, the desorption time is 5min, the desorption flow is 50mL/min, and no flow distribution is realized; the cold trap analysis temperature is 290 ℃, the flow rate is 20mL/min, and the split ratio is 6: 1.
4. Gas chromatography-mass spectrometry combined analysis condition
Chromatographic conditions are as follows: capillary chromatography column HP-5ms Ultra Inert,325 ℃ (30m × 250 μm × 0.25 μm); temperature programming: setting the initial column incubator at 40 deg.C, maintaining for 0min, heating to 88 deg.C at 3 deg.C/min, maintaining for 0min, then heating to 290 deg.C at 10 deg.C/min, and maintaining for 10 min; carrier gas: helium (99.999% pure); flow rate of carrier gas: 1.3 mL/min; and (3) sample introduction mode: no split-flow sample introduction.
Mass spectrum conditions: an ion source: electron impact ion source (EI source); electron energy: 70 eV; transmission line temperature: 280 ℃; ion source temperature: 230 ℃; temperature of the quadrupole rods: 150 ℃; the mass scanning range is 33-550 amu, the acquisition frequency is 50Hz, and the solvent delay is 2 min.
5. Qualitative and quantitative analysis of volatile components
Qualitative analysis is carried out through an automatic mass spectrum deconvolution qualitative system, the peak width of a deconvolution parameter is set to be 20, the requirements of resolution, sensitivity and chromatographic peak shape are set to be medium, the minimum value of a matching factor is set to be 80, NIST14 is selected for spectrogram retrieval, and meanwhile, auxiliary qualitative analysis is carried out by combining with literature. The quantitative analysis is calculated by a chemical workstation data processing system according to a peak area normalization method to obtain the relative percentage content of each component.
6. Qualitative and quantitative result analysis of tea aroma components
The total ion flow diagram of fragrance components of West lake Longjing is shown in figure 1, and 96 chemical components (shown in table 2) are qualitatively obtained, wherein the chemical components mainly comprise 1 amine compound, 10 alcohol compounds, 1 azacyclo compound, 2 phenol compounds, 2 furan compounds, 8 aldehyde compounds, 47 hydrocarbon compounds, 6 ketone compounds and 19 ester compounds. From the percentage content, the hydrocarbon compound accounts for 39.68 percent at most, the nitrogen heterocyclic compound accounts for 23.83 percent at the next time, and the aldehyde compound, the alcohol compound, the ester compound, the ketone compound, the phenolic compound, the furan compound and the amine account for 11.98 percent, 10.34 percent, 10.11 percent, 2.29 percent, 1.04 percent, 0.69 percent and 0.03 percent respectively. From the individual compounds, some important components such as caffeine (23.83%), nonanal (4.47%), hexanal (3.95%), benzyl alcohol (2.84%), myristyl alcohol (2.46%), geraniol (1.19%), beta-ionone (1.02%) and the like are mainly present.
The total ion flow diagram of the aroma components of the camellia mongolica is shown in fig. 2, and 81 chemical components (shown in table 3) are qualitatively obtained, wherein the chemical components mainly comprise 13 alcohol compounds, 1 nitrogen heterocyclic compound, 2 phenol compounds, 1 furan compound, 4 aldehyde compounds, 35 hydrocarbon compounds, 6 ketone compounds and 19 ester compounds. From the percentage content, the highest alcohol compound content accounts for 37.52 percent, the second hydrocarbon compound content accounts for 28.87 percent, and the nitrogen heterocyclic compounds, ester compounds, ketone compounds, phenolic compounds, aldehyde compounds and furan compounds respectively account for 12.57 percent, 12.69 percent, 5.77 percent, 1.46 percent, 1.09 percent and 0.04 percent. From the individual compounds, there are mainly some important components, such as phenethyl alcohol (20.10%), caffeine (12.57%), benzyl alcohol (6.28%), dihydroactinidiolide (2.34%), jasmone (2.21%), -cadinene (2.08%), beta-ionone (2.00%), linalool (1.92%), myristyl alcohol (1.70%), geraniol (1.20%).
The aroma components of the West lake Longjing tea and the Mengding camellia tea are obviously different due to the difference of the tea varieties, the growth environment, the processing technology and other factors.
TABLE 2 qualitative and quantitative table for aroma components of West lake Longjing tea
TABLE 3 qualitative and quantitative table of aroma components of Mengding camellia
In conclusion, the micro-cell thermal extraction technology is combined with thermal desorption and gas chromatography-mass spectrometry technology to quickly and effectively analyze aroma components in the tea, so that a complex pretreatment process is avoided, an organic solvent is not required, the method is environment-friendly and safe, the sample consumption is small, the method is an ideal tea aroma detection means, and the method has a wide application prospect.
The above description is only an embodiment of the present invention, but the technical features of the present invention are not limited thereto, and any person skilled in the relevant art can change or modify the present invention within the scope of the present invention.
Claims (4)
1. A method for rapidly measuring tea aroma components is characterized by comprising the following steps:
(1) putting the weighing cups containing the tea samples to be tested into a micro-cell thermal extraction instrument in parallel, inserting adsorption tubes in parallel above the corresponding micro-cells, and continuously introducing high-purity carrier gas with a constant flow rate under a certain temperature condition to extract aroma components of the tea;
the sampling amount of a tea sample to be detected is 0.8-1.2 g, the temperature of the micro-pool heat extraction treatment is 75-85 ℃, and the time is 14-16 min;
the adsorption tube needs to be aged before use: introducing high-purity N2Placing the adsorption tubes into an aging instrument, keeping the flow rate in each adsorption tube at 50-100mL/min, adjusting the pressure to 15-30psi, and heating to 300-350 ℃ for 20-40 min;
(2) directly carrying out GC-MS detection after the sample adsorption tube subjected to the micro-cell thermal extraction treatment is thermally desorbed;
the thermal desorption conditions were: the Pre-purge time is 1min, the flow rate is 20mL/min, and the split ratio is 6: 1; the desorption temperature is 275 ℃, the desorption time is 5min, the desorption flow is 50mL/min, and no flow distribution is realized; the cold trap analysis temperature is 290 ℃, the flow rate is 20mL/min, and the split ratio is 6: 1;
the GC-MS conditions were:
chromatographic conditions are as follows: capillary chromatographic column HP-5ms Ultra insert, 325 ℃; temperature programming: setting the initial column incubator at 40 deg.C, maintaining for 0min, heating to 88 deg.C at 3 deg.C/min, maintaining for 0min, then heating to 290 deg.C at 10 deg.C/min, and maintaining for 10 min; carrier gas: helium gas; flow rate of carrier gas: 1.3 mL/min; and (3) sample introduction mode: no shunt sampling;
mass spectrum conditions: an ion source: electron bombardment ion source; electron energy: 70 eV; transmission line temperature: 280 ℃; ion source temperature: 230 ℃; temperature of the quadrupole rods: 150 ℃; the mass scanning range is 33-550 amu, the acquisition frequency is 50Hz, and the solvent delay is 2 min.
2. The method according to claim 1, wherein the sample amount of the tea leaves to be tested is 1g, and the temperature of the micro-cell thermal extraction treatment is 80 ℃ for 15 min.
3. The method according to claim 1, wherein 6 weighing cups containing tea samples are placed in 6 micro-cells in parallel, adsorption tubes are inserted into the upper space of the corresponding micro-cells side by side, the pressure of each adsorption tube is adjusted to 35-45 psi, the adsorption tubes are pulled out and sealed after purging for 5-20 min by using high-purity nitrogen as carrier gas.
4. The method of claim 1, wherein the GC-MS detection result is qualitatively analyzed by an automatic mass spectrum deconvolution qualitative system, and the quantitative analysis is calculated by a peak area normalization method to obtain the relative percentage content of each component.
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Application publication date: 20180327 Assignee: Foshan Aiweidun Technology Co.,Ltd. Assignor: TEA Research Institute CHINESE ACADEMY OF AGRICULTURAL SCIENCES Contract record no.: X2023980032397 Denomination of invention: A method for rapid determination of aroma components in tea Granted publication date: 20200908 License type: Common License Record date: 20230222 |