阅读说明：本技术 薄荷清新茶指纹图谱的检测方法 (Method for detecting fingerprint of mint refreshing tea ) 是由 金俊杰 李国维 刘爱国 于 2021-07-31 设计创作，主要内容包括：本发明公开了薄荷清新茶指纹图谱的检测方法,它包括步骤1、薄荷清新茶供试品溶液的制备；步骤2、混合对照品溶液的制备：步骤3、分别精密吸取混合对照品溶液和供试品溶液注入液相色谱仪,记录色谱图；步骤4,薄荷清新茶指纹图谱仪器导出,导入中药色谱指纹图谱相似度评价系统,选择不同批次薄荷清新茶的色谱图中均存在的色谱峰作为共有峰；用平均值计算法生成薄荷清新茶的对照指纹图谱；计算各共有峰的相对保留时间、相对峰面积；将薄荷清新茶指纹图谱和混合标准品图谱进行比对,指认主要成分峰。本发明所提供的薄荷清新茶指纹图谱,能全面,客观地表征薄荷清新茶的质量。且检测方法具有方法简便、稳定、精密度高、重现性好等优点。(The invention discloses a method for detecting fingerprint of mint refreshing tea, which comprises the following steps of 1, preparing a test solution of the mint refreshing tea; step 2, preparation of a mixed reference solution: step 3, respectively and precisely sucking the mixed reference solution and the test solution to be injected into a liquid chromatograph, and recording a chromatogram; step 4, guiding out the fingerprint instrument of the mint fresh tea, guiding the fingerprint instrument into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and selecting chromatographic peaks existing in the chromatograms of different batches of mint fresh tea as common peaks; generating a comparison fingerprint of the mint fresh tea by using an average value calculation method; calculating the relative retention time and the relative peak area of each common peak; comparing the fingerprint of the mint fresh tea with the fingerprint of the mixed standard substance, and identifying the peak of the main component. The fingerprint spectrum of the mint refreshing tea provided by the invention can comprehensively and objectively represent the quality of the mint refreshing tea. And the detection method has the advantages of simple and stable method, high precision, good reproducibility and the like.)

1. A method for detecting fingerprint spectrum of mint refreshing tea is characterized by comprising the following steps:

step 1, preparing a test solution of the mint refreshing tea:

weighing fresh mint tea samples of different batches respectively, pulverizing, placing in a round-bottomed bottle, adding methanol solution, performing ultrasonic extraction, filtering, placing filtrate in a volumetric flask, adding methanol to a constant volume, and filtering with a 0.45 μm microporous membrane to obtain a sample solution;

step 2, preparation of mixed reference solution:

precisely weighing chlorogenic acid, luteolin and quercetin reference substances, placing in a volumetric flask, adding methanol to desired volume to scale, shaking, and making into mixed reference substance solution;

step 3, precisely absorbing the test solution in the step 1 and the reference solution in the step 2 respectively, injecting the test solution and the reference solution into a high performance liquid chromatograph, and recording a chromatogram;

step 4, exporting the fingerprint of the mint fresh tea test solution obtained in the step 3, and importing the fingerprint into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; selecting chromatographic peaks existing in chromatograms of different batches of mint fresh tea as common peaks; generating a control fingerprint of the mint refreshing tea by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; marking chemical components of peaks in the comparison fingerprint spectrum according to the retention time of the mixed reference solution chromatogram;

and 5, comparing the fingerprint spectrum of the mint refreshing tea obtained in the step 3 with the spectrum of the mixed standard substance, and identifying that the No. 1 peak in the mint refreshing tea is chlorogenic acid, the No. 2 peak is luteoloside and the No. 7 peak is quercetin.

2. The method for detecting the fingerprint of the mint refreshing tea according to claim 1, wherein the preparation method of the mint refreshing tea test solution in the step 1 comprises the following steps: taking 5g of 8 batches of fresh mint tea samples, pulverizing, placing in a round-bottom flask, carrying out ultrasonic extraction, filtering, placing filtrate in a 100mL volumetric flask, adding methanol to a constant volume, and filtering through a 0.45 mu m microporous filter membrane to obtain a test solution.

3. The method for detecting fingerprint of mint refreshing tea according to claim 1, wherein the step 2 of preparing the mixed reference solution comprises the following steps: precisely weighing chlorogenic acid, luteolin and quercetin reference substances, placing in a volumetric flask, adding methanol to constant volume to scale, shaking, and making into mixed reference substance solution containing 0.1019mg/mL chlorogenic acid, 0.1001mg/mL luteolin and 0.0529mg/mL quercetin.

4. The method for detecting the fingerprint of the mint refreshing tea according to claim 1, wherein in the step 3, the liquid chromatography conditions are as follows: a chromatographic column: purospher STAR LP RP-18 endclamped, mobile phase: acetonitrile and 0.1% phosphoric acid water, gradient elution, diode array detector, detection wavelength: 245nm, column temperature 30 ℃, flow rate 1.0mL/min, sample injection volume: 10 μ L, gradient elution procedure as follows:

5. the method for detecting the fingerprint of the mint refreshing tea as claimed in claim 1, wherein the fingerprint contains 7 peaks, the retention time of the 7 peaks is 20.464min for peak 1, 53.76min for peak 2, 55.59min for peak 3, 62.323min for peak 4, 66.745min for peak 5, 71.043min for peak 6 and 105.092min for peak 7, wherein the 1 peak is chlorogenic acid, the 2 peak is luteolin and the 7 peak is quercetin.

6. The method for detecting the fingerprint of the mint refreshing tea according to any one of claims 1 to 5, wherein the mint refreshing tea comprises the following medicinal ingredients in parts by weight: 4-8 parts of mint, 2-4 parts of honeysuckle, 2-4 parts of chrysanthemum, 5-10 parts of hawthorn, 5-10 parts of momordica grosvenori and 2-4 parts of lophatherum gracile.

7. The method for detecting the fingerprint of the mint refreshing tea as claimed in claim 6, wherein the mint refreshing tea comprises the following medicinal ingredients in parts by weight: 4 parts of mint, 2 parts of honeysuckle, 2 parts of chrysanthemum, 5 parts of hawthorn, 5 parts of momordica grosvenori and 2 parts of lophatherum gracile.

Technical Field

The invention relates to a detection method of traditional Chinese medicine substituted tea, in particular to a detection method of fingerprint spectrum of mint refreshing tea.

Background

The traditional Chinese medicine substituted tea is a special traditional Chinese medicine preparation formulation for treating diseases, conditioning, building body and prolonging life in traditional Chinese medicine, and plays an important role in the medical health care industry. The traditional Chinese medicine substitutive tea can be dialectically formulated according to the disease condition, can be increased or decreased according to the disease condition, can be properly used according to the performance characteristics of the medicine, and the like, has simple program, convenient preparation and strong pertinence, not only keeps the characteristics of flexible increase and decrease and remarkable curative effect of dialectical treatment of the traditional Chinese medicine decoction, but also overcomes the defects of complicated decoction, inconvenient carrying and the like of the traditional decoction, and is suitable for the development trend of accelerated pace of modern life. The traditional Chinese medicine substituted tea is convenient to store and carry, can be drunk for many times at any time, is completely absorbed, can be used for preparing special conditions or certain emergencies, and has good auxiliary treatment effect. From the view of the property of the medicine, the medicine of the traditional Chinese medicine substitutional tea is mild in property, does not hurt the stomach, is sweet in taste or slightly bitter and cold in taste, has the functions of disease removal and conditioning, is tasteless and bitter, and is particularly suitable for children patients. The Chinese medicinal tea is suitable for frequent administration with mild property and no damage to stomach qi, so it can be administered for a long time to relieve the symptoms of viscera, and is especially suitable for treating chronic diseases and regulating body function.

From the perspective of traditional Chinese medicine, the mint refreshing tea aims at clearing heat and purging fire, soothing liver and regulating qi, takes heat-clearing and fire-purging medicinal materials as main materials, and is supplemented with medicines for cooling blood, dissolving turbidity, tonifying spleen and tonifying qi to promote qi and invigorate qi. Herba Menthae has effects of inducing sweat, relieving fever, dispersing stagnated liver qi, relieving sore throat, relieving pain, and relieving itching; flos Lonicerae has effects of clearing away heat and toxic materials, dispelling pathogenic wind and heat, cooling blood, and relieving dysentery; the chrysanthemum has the effects of dispelling wind, clearing heat, calming liver, improving eyesight, clearing heat and removing toxicity; the hawthorn can promote digestion, invigorate stomach, promote qi circulation, remove blood stasis, eliminate turbid pathogen and reduce blood fat; fructus Siraitiae Grosvenorii has effects of clearing heat, moistening lung, relieving sore throat, nourishing kidney, moistening lung, invigorating spleen, and invigorating qi; lophatherum gracile has the effects of clearing heat, purging fire, relieving restlessness and promoting urination; experiments show that the health-care tea has good effects of clearing heat and purging fire, promoting qi circulation and removing blood stasis, nourishing kidney and moistening lung and the like when being drunk daily.

At present, the quality detection method of the mint refreshing tea is less. The invention adopts the high performance liquid chromatography to establish the fingerprint spectrum detection method of the mint refreshing tea, and has important significance for ingredient identification, quality evaluation and quality standard formulation of the mint refreshing tea.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to overcome the defects of the prior art and provides the fingerprint detection method of the mint refreshing tea, which can objectively, comprehensively and accurately evaluate the quality of the mint refreshing tea and has important significance for controlling the quality of the mint refreshing tea and ensuring the clinical curative effect.

The technical scheme is as follows: in order to achieve the purpose, the invention adopts the technical scheme that:

a method for detecting fingerprint spectrum of mint refreshing tea is characterized by comprising the following steps:

step 1, preparing a test solution of the mint refreshing tea:

precisely weighing mint fresh tea samples of different batches respectively, powdering, placing in a round-bottomed bottle, adding methanol solution, reflux extracting, filtering, placing the filtrate in a volumetric flask, adding methanol to a constant volume, and filtering with a 0.45 μm microporous membrane to obtain a sample solution;

step 2, preparation of mixed reference solution:

precisely weighing chlorogenic acid, luteolin and quercetin reference substances, placing in a volumetric flask, adding methanol to desired volume to scale, shaking, and making into mixed reference substance solution;

step 3, precisely absorbing the test solution in the step 1 and the reference solution in the step 2 respectively, injecting the test solution and the reference solution into a high performance liquid chromatograph, and recording a chromatogram;

step 4, exporting the fingerprint of the mint fresh tea test solution obtained in the step 3, and importing the fingerprint into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; selecting chromatographic peaks existing in chromatograms of different batches of mint fresh tea as common peaks; generating a control fingerprint of the mint refreshing tea by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; marking chemical components of peaks in the comparison fingerprint spectrum according to the retention time of the mixed reference solution chromatogram;

and 5, comparing the fingerprint spectrum of the mint refreshing tea obtained in the step 3 with the spectrum of the mixed standard substance, and identifying that the No. 1 peak in the mint refreshing tea is chlorogenic acid, the No. 2 peak is luteoloside and the No. 7 peak is quercetin.

As a preferred scheme, in the above method for detecting fingerprint of mentha arvensis fresh tea, step 1, the method for preparing the test solution of mentha arvensis fresh tea comprises the following steps: taking 5g of fresh mint tea sample powder of 8 batches, placing the fresh mint tea sample powder in a 100mL round-bottomed bottle, adding 50mL of methanol solution, carrying out ultrasonic extraction for 30min, filtering, placing the filtrate in a 100mL volumetric flask, adding methanol to a constant volume, and filtering through a 0.45-micrometer microporous membrane to obtain a sample solution.

Preferably, the method for detecting fingerprint of mint refreshing tea comprises the following steps of 2, preparing a mixed reference solution: precisely weighing chlorogenic acid, luteolin and quercetin reference substances, placing in a volumetric flask, adding methanol to constant volume to scale, shaking, and making into mixed reference substance solution containing 0.1019mg/mL chlorogenic acid, 0.1001mg/mL luteolin and 0.0529mg/mL quercetin.

Preferably, in the method for detecting fingerprint of mentha arvensis fresh tea, in step 3, the liquid chromatography conditions are as follows: a chromatographic column: purospher STAR LP RP-18 endclamped, mobile phase: acetonitrile and 0.1% phosphoric acid water, gradient elution, diode array detector, detection wavelength: 245nm, column temperature 30 ℃, flow rate 1.0mL/min, sample injection volume: 10 μ L, gradient elution procedure as follows:

Procedure	time (a)min）	Acetonitrile volume (%)
			1	0	5
2	10	10
			3	20	12
4	30	13
			5	54	19
6	99	22
			7	105	30
8	125	50

Preferably, in the method for detecting fingerprint of mint fresh tea, 7 peaks are shared in the fingerprint.

Optimizing fingerprint detection conditions:

1. in the aspect of preparation optimization of sample solution

According to the invention, through experimental comparison of different extraction methods (ultrasonic extraction, reflux extraction, percolation extraction and the like) and different extraction solvents (methanol, water, 50% ethanol water solution, 70% ethanol, 95% ethanol and absolute ethanol), the results show that the spectrogram difference component obtained by ultrasonic extraction is relatively comprehensive, the extraction efficiency is high, the separation degree is good, so that the ultrasonic extraction method is adopted; in the investigation of the extraction solvent, the chromatogram of the methanol extract is found to have the most information content and the highest component content, so that methanol is selected for extraction.

2. In the aspect of optimizing chromatographic conditions

According to the invention, a diode array detector is adopted to inspect the detection wavelength, chromatograms at positions of 205nm, 225nm, 245nm and 280nm are extracted, and when the detection wavelength is found to be 245nm, the information content contained in the chromatograms is most comprehensive and the base line is stable, so 245nm is selected as the detection wavelength;

the invention screens the flow rate (1 mL/min, 0.9mL/min, 0.8mL/min, 0.6 mL/min), and finally selects the gradient condition with the flow rate of 1.0mL/min because the components in the mint refreshing tea have better peak shape and better separation effect under the flow rate of 1 mL/min.

The invention compares the elution effects of 5 different elution systems of methanol-water, acetonitrile-0.1% formic acid, acetonitrile and 0.05% phosphoric acid water, and acetonitrile-0.1% phosphoric acid water under different gradients. As a result, it was found that acetonitrile and 0.1% phosphoric acid water were finally selected as the mobile phase because the separation effect of each component in the peppermint refreshing tea was good when acetonitrile and 0.1% phosphoric acid water were used as the mobile phase.

After the optimal fluidity composition is determined, the optimal gradient elution procedure is screened through a large number of experiments, and experiments show that when acetonitrile is adopted for 0-10 min, the volume is 5% -10%; 10-20 min acetonitrile with volume of 10-12%; the volume of acetonitrile is 12-13% in 20-130 min; 30-54 min acetonitrile with volume of 13-19%; the volume of acetonitrile is 19-22% in 54-99 min; the volume of acetonitrile is 22-30% in 99-105 min; acetonitrile with the volume of 30-50% in 105-125 min; good separation degree of each spectrum peak in the fingerprint spectrum can be realized.

The fingerprint spectrum detection method of the mint refreshing tea comprises the following medicinal components in parts by weight: 4-8 parts of mint, 2-4 parts of honeysuckle, 2-4 parts of chrysanthemum, 5-10 parts of hawthorn, 5-10 parts of momordica grosvenori and 2-4 parts of lophatherum gracile.

Preferably, the method for detecting the fingerprint of the mint refreshing tea is characterized in that the mint refreshing tea comprises the following medicinal components in parts by weight: 4 parts of mint, 2 parts of honeysuckle, 2 parts of chrysanthemum, 5 parts of hawthorn, 5 parts of momordica grosvenori and 2 parts of lophatherum gracile.

Has the advantages that:

1. according to the structural property characteristics of active ingredients contained in the mint refreshing tea, the optimal mobile phase composition is screened out through a large number of experiments, and analysis conditions such as gradient elution procedures, flow rate, detection wavelength, chromatographic column, column temperature and the like are verified through a plurality of experiments.

2. The fingerprint spectrum detection method of the mint refreshing tea, provided by the invention, can comprehensively, objectively and accurately detect and evaluate the quality of the mint refreshing tea, and has important significance for ensuring the curative effect of the mint refreshing tea.

3. The fingerprint of the mint refreshing tea established by the method provided by the invention can effectively represent the quality of the mint refreshing tea, objectively reflect the front and back sequence and the mutual relation of each fingerprint characteristic peak, pay attention to the overall facial features, avoid the one-sidedness of the quality of the mint refreshing tea determined by measuring individual chemical components, and reduce the possibility of manual treatment for reaching the quality standard.

4. The method for detecting the fingerprint of the mint refreshing tea provided by the invention has the advantages of simplicity, convenience, good stability, high precision, good reproducibility and the like.

Drawings

FIG. 1 is a chromatogram of a mixed control according to the present invention.

Fig. 2 is a control fingerprint of a sample of mint fresh tea of the present invention.

FIG. 3 shows the fingerprint spectrum of 8 batches of test samples of the mint fresh tea sample of the invention.

Detailed Description

Embodiments of the present invention will be described in detail with reference to examples, in which specific conditions are not specified, according to conventional conditions or conditions recommended by manufacturers. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

The following examples used the following instruments and reagents:

experimental equipment

1.1 instruments

A high-performance liquid chromatography system with full-wave-band scanning (200- & 800 nm) of Shimadzu corporation in Japan comprises a full-automatic online degassing system, a full-automatic sample introduction system promience SIL-20A, a diode array detector SPD-M20A, an automatic temperature control column oven CTO-20A, a KQ3200DB type numerical control ultrasonic cleaner (ultrasonic instruments Co., Ltd., Kunshan city) and a BP121S electronic analysis balance (SARTORIUS).

1.2 drugs and reagents

The mint fresh tea sample is from Jiangsu Haihong pharmaceutical Co., Ltd; chlorogenic acid reference (batch No. 110753-201314) was purchased from China food and drug Biometrics institute; the luteolin control (batch No. 111720-201609) was purchased from China pharmaceutical and biological products institute; the quercetin reference product 100081-201610 (batch number) is purchased from China pharmaceutical biologicals institute; methanol (analytically pure); phosphoric acid (analytically pure); acetonitrile (chromatographically pure); water (ultrapure water).

Embodiment 1 a method for detecting fingerprint of mint fresh tea, comprising the following steps:

step 1, preparing a test solution of the mint refreshing tea:

taking 20g of 8 batches of fresh mint tea samples (4 g of mint, 2g of honeysuckle, 2g of chrysanthemum, 5g of hawthorn, 5g of momordica grosvenori and 2g of lophatherum gracile), pulverizing, putting into a 1000mL round-bottomed bottle, adding 400mL of methanol solution, carrying out ultrasonic extraction for 30min, filtering, putting the filtrate into a 500mL volumetric flask, adding methanol to a constant volume, and filtering through a 0.45-micron microporous filter membrane to obtain a test solution;

step 2, preparation of mixed reference solution:

precisely weighing chlorogenic acid, luteolin and quercetin reference substances, placing in a volumetric flask, adding methanol to constant volume to scale, shaking, and making into mixed reference substance solution containing 0.1019mg/mL chlorogenic acid, 0.1001mg/mL luteolin and 0.0529mg/mL quercetin.

Respectively and precisely sucking 8 batches of mint fresh tea test solution and mixed reference solution, injecting the test solution and the mixed reference solution into a high performance liquid chromatograph, and recording a chromatogram; the liquid chromatography conditions were: a chromatographic column: purospher STAR LP RP-18 endclamped, mobile phase: acetonitrile and 0.1% phosphoric acid water, gradient elution, diode array detector, detection wavelength: 245nm, column temperature 30 ℃, flow rate 1.0mL/min, sample injection volume: 10 μ L, gradient elution procedure as follows:

Procedure	time (min)	Acetonitrile volume (%)
			1	0	5
2	10	10
			3	20	12
4	30	13
			5	54	19
6	99	22
			7	105	30
8	125	50

Step 4, exporting the fingerprints of the 8 batches of mint fresh tea test solution obtained in the step 3, and importing the fingerprints into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system 2012A; selecting chromatographic peaks existing in chromatograms of 8 batches of mint fresh tea as common peaks; generating a control fingerprint of 1 batch of mint refreshing tea by using an average value calculation method, and calculating the relative retention time and the relative peak area of each common peak; results 7 common peaks were found in 1 batch of raw mentha arvensis tea, the reference fingerprint is shown in FIG. 2, and the fingerprint of 8 batches of the test product is shown in FIG. 3. The retention time of chlorogenic acid is 20.464min, and the retention time of luteoloside is 53.76 min; the quercetin retention time is 105.092 min.

Step 5, comparing the fingerprint spectrum of the mint refreshing tea obtained in the step 3 with the spectrum of the mixed standard substance (figure 1), identifying the main components, and comparing the number 1, 2 and 7 chromatographic peaks in the mint refreshing tea to obtain the following components: chlorogenic acid (retention time 20.464 min), luteolin (retention time 53.76 min), and quercetin (retention time 105.092 min).

Meanwhile, the invention uses the automatically generated contrast HPLC fingerprint spectrum R to generate a common chromatographic peak mode, and the common chromatographic peaks of 8 batches of the mint fresh tea traditional Chinese medicines of Jiangsu Haoyang pharmaceutical manufacturers are analyzed and calculated to have relatively good similarity, which shows that the fingerprint spectrum established by the mint fresh tea traditional Chinese medicine established by the method can well detect the quality of multiple batches of the mint fresh tea in Jiangsu Haoyang pharmaceutical industries, and the results are shown in Table 1.

TABLE 1 similarity between batches of samples and consensus patterns

	S1	S2	S3	S4	S5	S6	S7	S8	R
										S1	1	0.905	0.926	0.935	0.908	0.944	0.927	0.911	0.935
S2	0.905	1	0.957	0.903	0.921	0.902	0.905	0.906	0.922
										S3	0.926	0.957	1	0.941	0.963	0.944	0.911	0.975	0.923
S4	0.935	0.903	0.941	1	0.922	0.989	0.999	0.901	0.917
										S5	0.908	0.921	0.963	0.922	1	0.942	0.939	0.925	0.966
S6	0.944	0.902	0.944	0.989	0.942	1	0.92	0.949	0.948
										S7	0.927	0.905	0.911	0.999	0.939	0.92	1	0.907	0.911
S8	0.911	0.906	0.975	0.901	0.925	0.949	0.907	1	0.952
										R	0.935	0.922	0.923	0.917	0.945	0.948	0.911	0.952	1

Example 2 methodological study of fingerprint detection

1. Methodology investigation

1.1 precision investigation

Taking 20g of fresh mint tea samples (4 g of mint, 2g of honeysuckle, 2g of chrysanthemum, 5g of hawthorn, 5g of momordica grosvenori and 2g of lophatherum gracile), preparing a sample solution according to the sample preparation method of the example 1, carrying out continuous sample injection for 6 times with the sample injection amount of 10 mu L each time, detecting according to the chromatographic conditions of the example 1, measuring an HPLC chromatogram, and inspecting 7 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 1.2%, the area RSD of the common fingerprint peaks is less than 2.4%, and the instrument precision is better.

1.2 stability Studies

Taking 20g of fresh mint tea samples (4 g of mint, 2g of honeysuckle, 2g of chrysanthemum, 5g of hawthorn, 5g of momordica grosvenori and 2g of lophatherum gracile), preparing a sample solution according to the sample preparation method of the example 1, injecting samples for 0, 2, 4, 8, 12 and 24 hours according to the chromatographic conditions in the example 1, recording a chromatogram, and inspecting 7 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 1.7%, the area RSD of the common peak is less than 2.1%, and the sample solution has good stability within 24 hours.

1.3 repeatability test

Taking 6 parts of fresh mint tea sample, preparing a sample solution according to the sample preparation method in the example 1, respectively measuring, recording a chromatogram, and inspecting 7 common fingerprint peaks in the chromatogram, wherein the results show that the retention time RSD of the common fingerprint peaks is less than 1.0 percent, and the area RSD of the common fingerprint peaks is less than 2.9 percent, which indicates that the method has good repeatability.

The experimental results show that the fingerprint spectrum detection method for the mint refreshing tea provided by the invention has the advantages of good stability, high precision and good repeatability, can comprehensively and objectively evaluate the quality of the mint refreshing tea, and has important significance for ensuring the clinical curative effect.

The above embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and the scope of the present invention is defined by the claims. Various modifications and equivalents may be made by those skilled in the art within the spirit and scope of the present invention, and such modifications and equivalents should also be considered as falling within the scope of the present invention.