CN117582475A - Compound eucommia ulmoides tea and preparation method, quality detection method and application thereof - Google Patents
Compound eucommia ulmoides tea and preparation method, quality detection method and application thereof Download PDFInfo
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- CN117582475A CN117582475A CN202311586550.1A CN202311586550A CN117582475A CN 117582475 A CN117582475 A CN 117582475A CN 202311586550 A CN202311586550 A CN 202311586550A CN 117582475 A CN117582475 A CN 117582475A
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- eucommia ulmoides
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- parts
- tea
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Classifications
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- A—HUMAN NECESSITIES
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- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/34—Tea substitutes, e.g. matè; Extracts or infusions thereof
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/355—Lonicera (honeysuckle)
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
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Abstract
The invention belongs to the field of foods or medicines, and particularly relates to compound eucommia ulmoides tea, a preparation method, a quality detection method and application thereof. The compound eucommia ulmoides tea is prepared from the following raw materials: eucommia ulmoides leaf, dendrobium flower, honeysuckle flower, grosvener siraitia and longan. The compound eucommia ulmoides tea is prepared according to the traditional Chinese medicine formula principle and the modern extraction process, the quality detection method is simple and accurate to operate, has good precision, reproducibility and stability, can provide basis for comprehensively evaluating and controlling the quality of the compound eucommia ulmoides tea, and has good oxidation resistance through the medicinal effect experiment, can be applied to health-care foods or medicines, and has wide application prospect.
Description
[ field of technology ]
The invention relates to the field of foods or medicines, in particular to compound eucommia ulmoides tea, a preparation method, a quality detection method and application thereof.
[ background Art ]
Oxidative damage is a damage to cells and tissues caused by an oxidative reaction. During normal metabolic processes, the body produces a certain number of reactive oxygen radicals, which have a certain physiological effect. However, when free radical generation and scavenging are unbalanced in the body, oxidative damage can occur. Oxidative damage is closely related to the occurrence and progression of a variety of diseases, including cardiovascular and cerebrovascular diseases, cancer, autoimmune diseases, aging, and the like.
Traditional Chinese medicine has some unique advantages in terms of antioxidation. First, traditional Chinese medicine emphasizes the concept of unity and focuses on regulating the overall function of the human body. The traditional Chinese medicine can strengthen the self-oxidation resistance of the organism by regulating the balance of qi and blood and yin and yang in the body. Secondly, the traditional Chinese medicine has various medicines which can provide natural antioxidant substances, such as baikal skullcap root, coptis root, red sage root and the like. The traditional Chinese medicines contain rich polyphenol compounds and antioxidase, and can neutralize free radicals and reduce oxidative damage. In addition, the traditional Chinese medicine also has various pharmacological activities such as anti-inflammatory and anti-tumor, and the like, and can comprehensively regulate the immunity and the antioxidation function of organisms.
In summary, there is a close relationship between oxidative damage and disease. Traditional Chinese medicine has some unique advantages in terms of antioxidation, including holistic concept, therapeutic methods for providing natural antioxidant substances and comprehensive regulatory functions. The traditional Chinese medicine plays an important role in antioxidation treatment and has important significance for preventing and treating diseases related to oxidative damage.
[ invention ]
In order to solve the technical problems, the invention provides the compound eucommia ulmoides tea, the preparation method, the quality detection method and the application thereof, wherein the compound eucommia ulmoides tea is prepared according to the traditional Chinese medicine formula principle and the modern extraction process.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a compound eucommia tea is prepared from the following raw materials: eucommia ulmoides leaf, dendrobium flower, honeysuckle flower, grosvener siraitia and longan.
Further, the material is prepared from the following raw materials in parts by weight: 3-6 parts of eucommia ulmoides leaves, 3-5 parts of dendrobium flowers, 2-4 parts of lonicera japonica, 1-3 parts of fructus momordicae and 1-3 parts of longan.
Preferably, the material is prepared from the following raw materials in parts by weight: 5 parts of eucommia ulmoides leaves, 4 parts of dendrobium flowers, 3 parts of lonicera confusa, 2 parts of fructus momordicae and 2 parts of longan.
A method for preparing the compound eucommia ulmoides tea comprises the following steps: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, decocting in water for 1-5 times, each time with water 10-30 times of the total weight of the raw materials, each time for 1-3 hr, filtering decoction, concentrating the filtrate to obtain extract, and drying.
Specifically, the method comprises the following steps: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, pulverizing into coarse powder, decocting with water for 3 times, adding water 20 times of the total weight of the raw materials for the first time, decocting for 2 hr, adding water 15 times of the total weight of the raw materials for the second and third times, decocting for 1 hr, filtering the decoction with filter cloth, evaporating the filtrate, concentrating to obtain extract, vacuum lyophilizing, pulverizing, and sieving with 60 mesh sieve.
The quality detection method of the compound eucommia ulmoides tea comprises the following steps:
(1) Chromatographic conditions: the chromatographic column is a C18 column, the mobile phase comprises a phase A and a phase B, the phase A is 0.1% phosphoric acid aqueous solution, the phase B is acetonitrile, and gradient elution is carried out; the ultraviolet detection wavelength is 327nm;
(2) Preparation of a control solution: precisely weighing chlorogenic acid reference substance, adding 50% methanol to obtain chlorogenic acid reference substance solution;
(3) Preparation of test solution: (1) preparation of the mixed solvent: tetrahydrofuran, methanol and 0.1mol/L hydrochloric acid are taken, and the volume ratio is 5:4:1, mixing to obtain the mixed solvent; (2) precisely weighing compound eucommia ulmoides tea powder, adding the mixed solvent, performing ultrasonic treatment, cooling, shaking uniformly, and filtering to obtain a sample solution;
(4) Assay: and (3) measuring according to the chromatographic conditions in the step (1) to obtain the product.
Further, the quality detection method comprises the following steps:
(1) Chromatographic conditions: the chromatographic column is a C18 column, the mobile phase comprises a phase A and a phase B, the phase A is 0.1% phosphoric acid water solution, the phase B is acetonitrile, and the gradient elution procedure is as follows: at 0-10min, the mobile phase A phase is 89% and the mobile phase B phase is 11%; at 10-30min, the phase A of the mobile phase is 89% -60% and the phase B of the mobile phase is 11% -40%; at 30-31min, the mobile phase A is 60-89% and the mobile phase B is 40-11%; at 31-35min, mobile phase A phase is 89% and mobile phase B phase is 11%; the flow rate is 1ml/min; the ultraviolet detection wavelength is 327nm; the column temperature is 30 ℃; the sample injection amount is 10 μl;
(2) Preparation of a control solution: taking 0.001g of chlorogenic acid reference substance, precisely weighing, placing into a 10ml volumetric flask, adding 50% methanol to scale, and preparing into chlorogenic acid reference substance solution;
(3) Preparation of test solution: (1) preparation of the mixed solvent: tetrahydrofuran, methanol and 0.1mol/L hydrochloric acid are taken, and the volume ratio is 5:4:1, mixing to obtain the mixed solvent; (2) precisely weighing 0.2g of compound eucommia tea powder, adding 100ml of the mixed solvent, weighing, performing ultrasonic treatment for 30min, cooling, weighing again, supplementing the reduced weight with the mixed solvent, shaking uniformly, and filtering to obtain a sample solution;
(4) Assay: and (3) measuring according to the chromatographic conditions in the step (1) to obtain the product.
Further, the chromatographic conditions of the step (1) are as follows: the instrument is an Shimadzu LC-2030plus high performance liquid chromatography analyzer; the column was a Diamond C18 column, the column size was 4.6 mm. Times.250 mm, 5. Mu.m.
The invention also provides the application of the compound eucommia ulmoides tea in preparing health-care food or medicine with the antioxidant function.
The invention also provides a preparation with an antioxidant function, which is prepared from the compound eucommia tea and a carrier acceptable in food or medicine.
In summary, due to the adoption of the technical scheme, the beneficial effects of the invention are as follows:
(1) The traditional Chinese medicine holds that yin-yang balance has close relation with antioxidation treatment. The balance of yin and yang can affect the antioxidant function of the organism. When yang qi is sufficient and yin qi is insufficient, a heat disease is likely to occur in the body, and the heat disease can increase the generation of oxidation reaction, so that the oxidative damage of cells is increased. When yin qi is sufficient and yang qi is insufficient, cold diseases easily occur in the body, and the cold diseases also affect the oxidation resistance of the body. Therefore, regulating yin-yang balance can maintain the antioxidant function of organism.
The compound eucommia ulmoides tea is prepared according to the traditional Chinese medicine formula principle, and the formula solution is as follows: eucommia ulmoides leaves have the effects of warming and nourishing liver and kidney and strengthening tendons and bones, can tonify yang qi of liver and kidney and strengthen physical energy, and are main medicines of the formula; dendrobium flower, flos Lonicerae and fructus Siraitiae Grosvenorii are all ministerial drugs, wherein the herba Dendrobii flower has effects of nourishing kidney and nourishing yin, can assist folium Eucommiae to tonify kidney yin, and flos Lonicerae has effects of clearing heat and detoxicating, cooling blood and stopping bleeding, can provide balance for yang tonifying effect of folium Eucommiae, prevent overheating, and fructus Siraitiae Grosvenorii has effects of nourishing kidney liquid, lowering fire and detoxicating, can coordinate tonifying effect of folium Eucommiae, and maintain balance of yin and yang; the monarch drug and the ministerial drug are nontoxic and drastic, so that the adjuvant drug is not needed; the longan has the effects of warming and invigorating spleen and stomach and tonifying qi and blood, and can be used as a guiding drug to strengthen the drug effect of the prescription and strengthen the effect of regulating yin and yang balance. The combination of the medicines is beneficial to keeping yin-yang qi-blood balance and vigorous function of viscera.
(2) The research results of in-vitro simulated digestion process show that the digestion products of the compound eucommia ulmoides tea have good antioxidant activity at each stage in the body, and the raw materials used in the compound eucommia ulmoides tea are both medicinal and edible raw materials, so that the compound eucommia ulmoides tea can be developed into a novel functional food or medicine, and the development requirement of the health industry is met.
(3) The determination method has the advantages of good separation effect, good peak shape, proper retention time, strong specificity, accurate determination result and stable determination method within a specified time. The method established in the research is reliable and accurate, is simple to operate, can ensure the quality and the curative effect of the product, and ensures the medication safety of patients.
[ description of the drawings ]
Fig. 1: high performance liquid chromatography of chlorogenic acid reference solution;
fig. 2: content determination chromatogram of sample solution (202302 batches);
fig. 3: the compound eucommia ulmoides tea powder of the embodiment 3 of the invention has the capability of scavenging hydroxyl free radicals (-OH);
fig. 4: the compound eucommia ulmoides tea powder of the embodiment 3 of the invention has superoxide anion free radical (-O) 2 - ) Cleaning ability;
fig. 5: the compound eucommia tea powder iron ion (Fe) of the embodiment 3 of the invention 3+ ) Reducing power.
In fig. 1-2, the chromatographic peaks correspond to: 1 is chlorogenic acid.
[ detailed description ] of the invention
Example 1
Prescription: 3g of eucommia ulmoides leaves, 3g of dendrobium flowers, 2g of lonicera confusa, 1g of momordica grosvenori and 1g of longan.
The preparation method comprises the following steps: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii, and arillus longan, pulverizing into coarse powder, adding 20 times of water, decocting for 3 hr, filtering the decoction with filter cloth, evaporating the filtrate, concentrating to obtain extract, vacuum freeze drying, pulverizing, and sieving with 60 mesh sieve.
Example 2
Prescription: 6g of eucommia ulmoides leaves, 5g of dendrobium flowers, 4g of honeysuckle flowers, 3g of momordica grosvenori and 3g of longan.
The preparation method comprises the following steps: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, pulverizing into coarse powder, decocting with water for 5 times, decocting with water 30 times of the total weight of the raw materials for 3 hr, decocting with water 20 times of the total weight of the raw materials for the second and third times, decocting with water 2 hr, decocting with water 10 times of the total weight of the raw materials for the fourth and fifth times, decocting for 1 hr, filtering the decoction with filter cloth, evaporating the filtrate, concentrating to obtain extract, vacuum freeze drying, pulverizing, and sieving with 60 mesh sieve.
Example 3
Prescription: 5g of eucommia ulmoides leaves, 4g of dendrobium flowers, 3g of honeysuckle flowers, 2g of momordica grosvenori and 2g of longan.
The preparation method comprises the following steps: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, pulverizing into coarse powder, decocting with water for 3 times, adding water 20 times of the total weight of the raw materials for the first time, decocting for 2 hr, adding water 15 times of the total weight of the raw materials for the second and third times, decocting for 1 hr, filtering the decoction with filter cloth, evaporating the filtrate, concentrating to obtain extract, vacuum lyophilizing, pulverizing, and sieving with 60 mesh sieve.
Test one: high performance liquid chromatography content determination 1.1 instrument and reagent for chlorogenic acid contained in compound eucommia ulmoides tea of embodiment 3 of the invention
Island liquid LC-2030plus high performance liquid chromatography system (shimadzu corporation, japan); analytical balance No. 00000246 by kyo (beijing company of the certolisco instruments); KQ-500 high frequency digital control ultrasonic cleaner (Kunshan ultrasonic instruments Co., ltd.).
Chlorogenic acid reference (110753-201817, content of 96.8%) of Chinese food and drug administration institute; methanol, acetonitrile, phosphoric acid (chromatographic purity); water (ultrapure water).
1.2 chromatographic conditions and System applicability
The column was a Diamond C18 column (4.6mm.times.250 mm,5 μm) and was eluted with a gradient of 0.1% aqueous phosphoric acid as mobile phase A and acetonitrile as mobile phase B according to Table 1. The flow rate is 1ml/min, the column temperature is 30 ℃, the sample injection amount is 10 μl, and the detection wavelength is 327nm. Under the chromatographic conditions, the theoretical plate number is not less than 10000 (as shown in FIG. 1) calculated as chlorogenic acid peak.
TABLE 1
1.3 preparation of solutions
1.3.1 preparation of control solution chlorogenic acid control 0.0010g was precisely weighed and placed in a 10ml volumetric flask, and 50% methanol was added to the scale to prepare chlorogenic acid control solution.
1.3.2 preparation of sample solution (1) preparation of mixed solvent: tetrahydrofuran, methanol and 0.1mol/L hydrochloric acid are taken, and the volume ratio is 5:4:1, mixing to obtain the mixed solvent; (2) precisely weighing 0.2g of compound eucommia tea powder, adding 100ml of the mixed solvent, weighing, performing ultrasonic treatment for 30min, cooling, weighing again, supplementing the reduced weight with the mixed solvent, shaking uniformly, and filtering to obtain the solution of the sample.
1.4 methodology investigation
1.4.1 Linear relation chlorogenic acid reference solutions 2, 4, 6, 8, 10, 12, 14 and 16. Mu.l were precisely taken and sampled and analyzed according to the above chromatographic conditions. Carrying out linear regression on the peak area integral value (Y) by using the sample injection quantity (X, mug), and calculating the linear range of chlorogenic acid to obtain a linear equation: y= 8308882.330x-100836.286, r=0.9998 (n=8), and the result shows that the sample injection amount is between 0.1936 and 1.5488 μg, and the sample injection amount has good linear relation with the peak area.
1.4.2 precision taking control solution, continuously sampling for 6 times according to the above chromatographic condition method, and determining chlorogenic acid peak area. The RSD of the chlorogenic acid peak area obtained by calculation is 0.84% in sequence, which shows that the precision of the used instrument is good.
1.4.3 stability the same sample solution was taken and assayed by the chromatographic conditions described above at 0, 2, 4, 6, 8, 10, 12 and 24h respectively, the liquid chromatographic peak area was recorded and the RSD for chlorogenic acid peak area was calculated to be 1.53%, indicating that the sample solution was stable within 24 h.
1.4.4.4 repeated accurate weighing of the same batch of samples, preparing solution according to the preparation method of the sample, measuring according to the chromatographic condition method, calculating RSD of chlorogenic acid area in the samples to be 1.22%, and indicating that the method has good repeatability.
1.4.5 sample recovery rate 6 portions of compound eucommia tea powder with known content are precisely weighed, about 0.10g of compound eucommia tea powder is divided into each portion, a certain amount of chlorogenic acid standard substance is respectively added, a solution is prepared according to a sample preparation method, the content is measured according to the chromatographic condition method, and the result is shown in table 2. The average recovery rate of chlorogenic acid is 99.06% and RSD is 1.27%, which shows that the sample recovery rate of the measuring method meets the requirement.
TABLE 2
1.5 determination of samples
3 batches of samples (prepared according to the prescription and the preparation method of example 3) were taken, solutions were prepared according to the preparation method of the test sample, and the chlorogenic acid content was measured according to the above chromatographic condition method, so that the chlorogenic acid content in samples of batches 202301, 202302 and 202303 was 2.48%, 2.35% and 2.43%, respectively, and fig. 2 is a content chromatogram of batch 202302.
Experiment two Compound eucommia ulmoides tea antioxidant activity study (in vitro simulated digestion method) of example 3 of the invention
2.1 instruments and reagents
Evolution201 ultraviolet spectrophotometry (Thermo company); centrifuge5430R Centrifuge (eppendorf corporation); BSA 124S-type electronic analytical balance (sartorius company, germany); HH-2 electric heating constant temperature water bath (national electric Co., ltd.); LGJ-10E vacuum freeze dryer (four-ring Furicke Instrument technologies Co.); JBQ-ZD8 all-temperature oscillator (Hengzhou Putian Instrument Co., ltd.).
The hydroxyl free radical (OH) determination kit, the inhibition and generation superoxide anion free radical (O2) determination kit and the total antioxidant capacity (T-AOC) determination kit are purchased from Nanjing to build biotechnology Co-Ltd; alpha-amylase, saccharifying enzyme, trypsin, pepsin, ascorbic acid, ammonium carbonate, magnesium chloride, cholic acid, sodium bicarbonate, sodium chloride, potassium dihydrogen phosphate, anhydrous calcium chloride and the like are all commercially available analytical purity.
2.2 preparation of in vitro simulated digestive juice
Weighing each reagent according to the table 3, respectively dissolving in a certain volume of pure water, preparing mother solution with corresponding concentration, and refrigerating for standby. Preparing simulated digestive juice, adding corresponding mother liquor according to the following table, adding distilled water to constant volume to 400ml, and refrigerating for standby.
TABLE 3 Table 3
Note that: caCl2 (H2O) 2 is mixed with other solutions to generate precipitation, so that the mixture is added before starting the next experiment; 2.
HCL or NaOH is used to adjust the corresponding pH.
2.3 in vitro simulation of digestion Process
2.3.1 simulation of oral digestion process:
3 parts of the sample are weighed, 0.5g of each part is added with 4ml of artificial simulated saliva, 25 mu l of CaCl2 (H2O) 2 and 750U of alpha-amylase (which can be dissolved with water to prepare 1500U/ml, 0.5ml is taken when the sample is used), and the mixture is mixed, dissolved with pure water and fixed to 10ml. Sealing, placing into a constant temperature oscillator, and oscillating at 37deg.C for 10min to simulate oral cavity digestion. After the reaction is finished, centrifuging for 15min at 4500r/min, and obtaining supernatant as oral cavity digestive juice. Inactivating enzyme in water bath at 70deg.C, and refrigerating at 4deg.C; the other two parts are used in the next stomach digestion process without enzyme deactivation.
2.3.2 simulation of gastric digestion process:
after the last digestion phase, 8ml of gastric simulant working solution, 5. Mu.l of CaCl2 (H2O) 2 solution and 40000U of pepsin are added into the non-enzyme-inactivated oral cavity digestive juice. After adjusting the pH to 3.0 with HCl solution, distilled water was set to 20ml. Mixing, placing in a constant-temperature water bath shaker, shaking at 37deg.C in dark for 2 hr to simulate gastric digestion, collecting supernatant after reaction, inactivating enzyme in a water bath at 70deg.C, and storing in a refrigerator at 4deg.C; the other part is used for the next intestinal digestion process without enzyme deactivation.
2.3.3 simulation of intestinal digestion
After the last digestion stage, 8ml of small intestine digestion working solution, 40. Mu.l of CaCl2 (H2O) 2 solution, 4000U of trypsin, 0.6g of cholic acid powder, pH value adjusted to 7.0 by NaOH and distilled water volume fixed to 40ml are added into the stomach digestion solution without enzyme. After being evenly mixed, the mixture is placed on a constant-temperature water bath shaking table, shaking is carried out for 4 hours at 37 ℃ in a dark place, and the supernatant is taken and placed on a refrigerator at 4 ℃ for storage.
2.4 antioxidant experiment
2.4.1 grouping
Divided into 5 groups: the original drug control group (tea powder 0.5g dissolved in 10ml pure water), oral cavity digestion group, stomach digestion group, intestinal digestion group, and positive drug control group (ascorbic acid drug).
2.4.2 detection of antioxidant Activity (according to the kit requirements)
(1) Scavenging of hydroxyl radicals.
The measurement principle is as follows: fenton reaction produces hydroxyl radicals in an amount corresponding to H 2 O 2 Proportional to the ratio. After the electron acceptor is given, a red substance is formed by the development of the Grignard reagent, which develops color with OH - Is proportional to how much.
The protocol and reagent compositions (50 tubes/48 samples) are as described in Table 4:
TABLE 4 Table 4
Note that: the preparation of the reagent is shown in the specification of the hydroxyl radical assay kit.
Inhibition ability = (control-a assay)/(a standard-a blank) ×c standard/V sample×n
C standard: standard concentration 8.824mmol/L; n: a sample pre-dilution multiple;
OH - clearance = (sample a-blank)/(control a-blank) ×100%
The measurement results are shown in Table 5:
TABLE 5
| Sample name | Absorbance A | Inhibition ability | OH-clearance |
| Blank pipe | -0.003 | ||
| Standard tube | 0.102 | ||
| Control tube | 0.814 | ||
| Measuring tube ascorbic acid medicine (15 mg) | 0.190 | 262.1988 | 23.6230 |
| Measuring tube tea powder raw medicine | 0.329 | 203.7923 | 40.6364 |
| Oral cavity digestion group for measuring tube tea powder | 0.510 | 127.7379 | 62.7906 |
| Stomach digestion group for measuring tube tea powder | 0.643 | 71.8525 | 79.0697 |
| Determination of tube tea powder sausage digestion group | 0.120 | 291.6121 | 15.0550 |
The higher the absorbance, the weaker the inhibition ability, and the absorbance A of each sample was measured at a concentration of 0.5mg/ml as shown in Table 5.
OH-clearance determination results: the OH-clearance of each component was determined as shown in FIG. 3. Stomach digestion group > oral digestion group > original drug > ascorbic acid drug > intestinal digestion group.
(2) Scavenging of superoxide anion radical:
the measurement principle is as follows: the simulated organism xanthine and Huang Piao make the oxidase reaction system produce superoxide anion free radical, and then add electron transfer material and color-developing agent to make it appear purple red, and can use spectrophotometer to measure absorbance. When the tested sample contains the superoxide anion radical inhibitor, the absorbance of the measuring tube is lower than that of the control tube; when the sample contains a substance that generates superoxide anion radical, the absorbance of the measurement tube is higher than that of the control tube. And (3) taking vitamin C as a standard, and calculating the influence capability of the detected object on the superoxide anion free radical.
The operation table is shown in table 6:
TABLE 6
Note that: the preparation of the reagent is shown in the specification of the determination kit for inhibiting and generating superoxide anion free radicals.
Anti-superoxide anion capacity = (control-a assay)/(control-a standard) ×c standard =1000×n
C standard: standard concentration 0.15mg/ml; n: dilution fold before sample testing.
O 2- Clearance = (a blank-a sample + a standard)/a blank%
The measurement results are shown in Table 7:
TABLE 7
| Sample name | Absorbance A | Resistance to superoxide anions | O 2- Clearance rate of |
| Standard tube | 0.118 | ||
| Control tube | 0.738 | ||
| Measuring tube tea powder raw medicine | 0.659 | 19.1129 | 26.6937 |
| Oral cavity digestion group for measuring tube tea powder | 0.615 | 29.7580 | 32.6558 |
| Stomach digestion group for measuring tube tea powder | 0.680 | 14.0322 | 23.8482 |
| Determination of tube tea powder sausage digestion group | 0.799 | -14.7581 | 7.72357 |
The higher the absorbance, the weaker the resistance to superoxide anions, and the absorbance A of each sample was measured at a concentration of 0.5mg/ml as shown in Table 7.
Anti-superoxide anion assay results: the anti-superoxide anion capacity of each component was measured, as shown in fig. 4, oral cavity digestive group > original drug > stomach digestive group > intestine digestive group.
(3) Determination of Total antioxidant Capacity
The measurement principle is as follows: there are many antioxidant substances in the body, which have the effect of adding Fe 3+ Reduction to Fe 2+ Is provided). When Fe < 2+ > and phenanthroline substances form a firm complex, the oxidation resistance of the complex can be measured by a colorimetric method.
Two reagent compositions and configurations:
the operation table is shown in table 8:
TABLE 8
Note that: the preparation of the reagent is shown in the specification of the total antioxidant capacity determination kit.
Anti-superoxide anion capacity = (assay-a control)/0.01/T x V inverse total/V sample x N
V inverse total: the total volume of the reaction system; n: a sample pre-dilution multiple;
the measurement results are shown in Table 9:
| sample name | Measuring tube absorbance | Absorbance of control tube | Total antioxidant capacity |
| Tea powder raw medicine | 1.123 | 0.311 | 100.1466 |
| Oral cavity digestion group for measuring tube tea powder | 1.204 | 0.228 | 120.3733 |
| Determination of tube tea powder gastric digestionGroup of | 0.151 | 0.028 | 15.17 |
| Determination of tube tea powder sausage digestion group | 0.152 | 0.028 | 15.2933 |
The higher the absorbance, the better the oxidation resistance, and the stronger the reducing power, and the absorbance A of each sample was measured at a concentration of 0.5mg/ml as shown in Table 9.
The total antioxidant capacity measurement results are shown in FIG. 5: oral cavity digestive group > tea powder raw medicine > intestinal digestive group > stomach digestive group.
2.5 conclusion
The research results of in vitro simulated digestion process show that the tea powder has good antioxidant activity in digestion products of various stages in the body, can be developed into a novel functional food, and meets the development requirements of the health industry.
The foregoing description is directed to the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the invention, and all equivalent changes or modifications made under the technical spirit of the present invention should be construed to fall within the scope of the present invention.
Claims (10)
1. The compound eucommia ulmoides tea is characterized by being prepared from the following raw materials: eucommia ulmoides leaf, dendrobium flower, honeysuckle flower, grosvener siraitia and longan.
2. The compound eucommia ulmoides tea according to claim 1, which is prepared from the following raw materials in parts by weight: 3-6 parts of eucommia ulmoides leaves, 3-5 parts of dendrobium flowers, 2-4 parts of lonicera japonica, 1-3 parts of fructus momordicae and 1-3 parts of longan.
3. The compound eucommia ulmoides tea according to claim 1, which is prepared from the following raw materials in parts by weight: 5 parts of eucommia ulmoides leaves, 4 parts of dendrobium flowers, 3 parts of lonicera confusa, 2 parts of fructus momordicae and 2 parts of longan.
4. A method for preparing the compound eucommia tea according to any one of claims 1 to 3, comprising the steps of: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, decocting in water for 1-5 times, each time with water 10-30 times of the total weight of the raw materials, each time for 1-3 hr, filtering decoction, concentrating the filtrate to obtain extract, and drying.
5. The method of compound eucommia tea according to claim 4, comprising the steps of: weighing folium Eucommiae, flos Dendrobii, flos Lonicerae, fructus Siraitiae Grosvenorii and arillus longan, pulverizing into coarse powder, decocting with water for 3 times, adding water 20 times of the total weight of the raw materials for the first time, decocting for 2 hr, adding water 15 times of the total weight of the raw materials for the second and third times, decocting for 1 hr, filtering the decoction with filter cloth, evaporating the filtrate, concentrating to obtain extract, vacuum lyophilizing, pulverizing, and sieving with 60 mesh sieve.
6. The quality detection method of the compound eucommia ulmoides tea according to any one of claims 1 to 3, which is characterized in that: the quality detection method comprises the following steps:
(1) Chromatographic conditions: the chromatographic column is a C18 column, the mobile phase comprises a phase A and a phase B, the phase A is 0.1% phosphoric acid aqueous solution, the phase B is acetonitrile, and gradient elution is carried out; the ultraviolet detection wavelength is 327nm;
(2) Preparation of a control solution: precisely weighing chlorogenic acid reference substance, adding 50% methanol to obtain chlorogenic acid reference substance solution;
(3) Preparation of test solution: (1) preparation of the mixed solvent: tetrahydrofuran, methanol and 0.1mol/L hydrochloric acid are taken, and the volume ratio is 5:4:1, mixing to obtain the mixed solvent; (2) precisely weighing compound eucommia ulmoides tea powder, adding the mixed solvent, performing ultrasonic treatment, cooling, shaking uniformly, and filtering to obtain a sample solution;
(4) Assay: and (3) measuring according to the chromatographic conditions in the step (1) to obtain the product.
7. The quality detection method of the compound eucommia ulmoides tea as defined in claim 6, which is characterized in that: the quality detection method comprises the following steps:
(1) Chromatographic conditions: the chromatographic column is a C18 column, the mobile phase comprises a phase A and a phase B, the phase A is 0.1% phosphoric acid water solution, the phase B is acetonitrile, and the gradient elution procedure is as follows: at 0-10min, the mobile phase A phase is 89% and the mobile phase B phase is 11%; at 10-30min, the phase A of the mobile phase is 89% -60% and the phase B of the mobile phase is 11% -40%; at 30-31min, the mobile phase A is 60-89% and the mobile phase B is 40-11%; at 31-35min, mobile phase A phase is 89% and mobile phase B phase is 11%; the flow rate is 1ml/min; the ultraviolet detection wavelength is 327nm; the column temperature is 30 ℃; the sample injection amount is 10 μl;
(2) Preparation of a control solution: taking 0.001g of chlorogenic acid reference substance, precisely weighing, placing into a 10ml volumetric flask, adding 50% methanol to scale, and preparing into chlorogenic acid reference substance solution;
(3) Preparation of test solution: (1) preparation of the mixed solvent: tetrahydrofuran, methanol and 0.1mol/L hydrochloric acid are taken, and the volume ratio is 5:4:1, mixing to obtain the mixed solvent; (2) precisely weighing 0.2g of compound eucommia tea powder, adding 100ml of the mixed solvent, weighing, performing ultrasonic treatment for 30min, cooling, weighing again, supplementing the reduced weight with the mixed solvent, shaking uniformly, and filtering to obtain a sample solution;
(4) Assay: and (3) measuring according to the chromatographic conditions in the step (1) to obtain the product.
8. The quality inspection method according to claim 6, wherein: the chromatographic conditions in the step (1) are as follows: the instrument is an Shimadzu LC-2030plus high performance liquid chromatography analyzer; the column was a Diamond C18 column, the column size was 4.6 mm. Times.250 mm, 5. Mu.m.
9. Use of the compound eucommia tea according to any one of claims 1 to 3 in the preparation of health food or medicine having an antioxidant function.
10. A preparation with an antioxidant function, which is characterized by being prepared from the compound eucommia tea according to any one of claims 1-3 and a carrier acceptable in food or medicine.
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