Hypoglycemic composition formula method based on large yellow tea and hypoglycemic composition

Abstract

The invention belongs to the technical field of plant medicine refining and improvement, and particularly relates to a formula method of a hypoglycemic composition based on large yellow tea and the hypoglycemic composition. According to the composition method, an ethanol water solution is adopted for extracting the Huangdan tea, GRA and PLSR are adopted for correlation analysis by means of the blood glucose change dynamic of an animal model and the change dynamic of blood entering components and metabolites after animals take drugs, the blood glucose reducing composition composition method based on the Huangdan tea is constructed, four active components are obtained, and the composition is prepared through a traditional Chinese medicine method. And the composition ratio of the four active components is adjusted and screened to obtain the hypoglycemic composition with a good glucose absorption inhibition effect.

Description

Technical Field

[0001] This invention belongs to the field of plant medicine refining and improvement technology, specifically relating to a formulation method of a hypoglycemic composition based on Huangda tea and a hypoglycemic composition. Background Technology

[0002] Tea has a long history of being both a food and a medicine. The Tang Dynasty medical text *Bencao Shiyi* states that "tea is a medicine for all diseases." Its various active ingredients give tea significant blood sugar-lowering potential, with mechanisms including reducing oxidative stress, enhancing pancreatic function, and regulating glucose and lipid metabolism. Huangda tea, produced in Huoshan, Anhui (Huoshan Huangda tea), is a lightly fermented yellow tea. It uses fresh leaves with stems ("one bud and three to five leaves") as raw material, undergoing a yellowing process followed by high-temperature roasting to develop its characteristic "yellow leaves and yellow liquor" and unique aroma. Studies have shown that Huangda tea is rich in catechins, flavonoids, and phenolic acids, exhibiting good effects in lowering blood sugar, regulating lipids, and improving metabolic syndrome.

[0003] Although considerable research has been conducted in this field on the hypoglycemic effect of Huangda tea, such as "The Influence of High-Temperature Roasting (Long-Fire) Process on the Chemical Composition and Hypoglycemic and Lipid-Lowering Activities of Huangda Tea" (Dissertation, Anhui Agricultural University) and "The Influence of Different Acidic Extraction Conditions on the Physicochemical Properties, Structure, and Hypoglycemic Activity of Huangda Tea Polysaccharides" (Xuan Kaili, Chen Hao, Wang Zhuang, et al., Food Science, 2025), the complexity of the effects of extraction and roasting processes, as well as the influence of methods for detecting blood-entry components, means that the hypoglycemic active substances in Huangda tea are not yet fully understood. This hinders the development of hypoglycemic drugs based on Huangda tea.

[0004] Therefore, establishing a formulation method for hypoglycemic compositions based on Huangda tea (i.e., a method for screening and combining active ingredients) and developing corresponding hypoglycemic compositions has significant economic and social value for the development of hypoglycemic drugs. Summary of the Invention

[0005] To address the above problems, the purpose of this invention is to provide a formulation method for a hypoglycemic composition based on Huangda tea, and a hypoglycemic composition. This invention includes the following technical solutions:

[0006] A method for formulating a hypoglycemic composition based on Huangda tea includes the following steps:

[0007] Preparation of S1 Huangda Tea Extract: Huangda tea was extracted with 70%~75% ethanol aqueous solution, and the extract was concentrated and the solvent was removed to prepare Huangda tea extract;

[0008] Screening of ingredients for S2 formulation: A type 2 diabetes animal model was established, and Huangda tea extract was administered by gavage. The area under the blood glucose-time curve Ai and the concentration or peak area of ​​the Huangda tea drug prototype and metabolites in the blood samples were measured in both the model animals and the treated animals. Gray-scale correlation analysis and partial least squares regression analysis were used to analyze the correlation between the difference in Ai between the model animals and the treated animals and the concentration or peak area of ​​the Huangda tea drug prototype and metabolites in the blood samples. Drug prototypes and / or metabolites with a gray correlation ≥ 0.8, a VIP value close to 1, and a negative correlation with the animal's blood glucose level were selected as ingredients for the formulation.

[0009] S3 Hypoglycemic Composition Formulation: Take the ingredients used in step S2 as active ingredients and combine them to prepare a hypoglycemic composition.

[0010] Preferably, in step S2, the concentration or peak area of ​​the original components and metabolites of Huangda Tea in the blood sample is determined by UPLC-Q-TOF / MS (ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry).

[0011] Preferably, the detection conditions of the UPLC-Q-TOF / MS include:

[0012] UPLC conditions: A C18 column was used as the stationary phase; column dimensions: 100 mm × 2.1 mm, 1.7 μm; mobile phase A was 0.1% formic acid in water, mobile phase B was acetonitrile, flow rate was 0.2 mL / min, and gradient elution program was as follows:

[0013] 0~5 min (95%A), 5~10 min (95%→90%A), 10~20 min (90%→84%A), 20~32 min (84%→80%A), 32~37 min (80%→65%A), 37~42 min (65%→45%A), 42~45 min (45%→30%A), 45~50min (30%→10%A), 50~53 min (10%A), 53~54 min (10%→95%A), 54~55 min (95%A).

[0014] Preferably, the detection conditions of the UPLC-Q-TOF / MS include:

[0015] Q / TOF-MS conditions: Electrospray ionization source was used for dual-mode detection (positive and negative ions). Ion source temperature was 120℃, scan range m / z 50-1200 Da, capillary voltage 3.0 kV (positive mode) / 2.5 kV (negative mode), cone voltage 40.0 kV, solvation temperature 350℃, solvent gas flow rate 600 L·h. -1Mass axis correction uses leucine-enkephalin, positive mode [M+H] + =556.2771, Negative Mode [MH] - =554.2615, MSE collision energy is 6V for low energy and 50~60V for high energy.

[0016] Preferably, in step S2, the type 2 diabetes animal model is a rodent model.

[0017] Preferably, in step S2, the area under the blood glucose-time curve Ai is calculated as follows: after modeling, the blood glucose values ​​at different time points are measured as G. i The different time points corresponding to the blood glucose collection are t i The area under the blood glucose-time curve is divided into multiple trapezoids using the trapezoidal method, and the area under the blood glucose-time curve is calculated as Ai.

[0018] .

[0019] Preferably, in step S3, the mass ratio of the active ingredients in the hypoglycemic composition is the ratio of the content of the corresponding active ingredients in the Huangda tea extract.

[0020] A hypoglycemic composition obtained by the aforementioned formulation method of a hypoglycemic composition based on Huangda tea, wherein the active ingredients of the hypoglycemic composition are: (2S,3R)-gallocatechin, (2S,3R)-Epigallocatechin, 3-p-Coumaroylquinic acid and Isoquercitrin.

[0021] The structural formula of the active ingredient is as follows:

[0022]

[0023] Preferably, the mass ratio of (2S,3R)-gallocatechin, (2S,3R)-Epigallocatechin, 3-p-Coumaroylquinic acid, and Isoquercitrin in the hypoglycemic composition is 1:(1.17~1.2):1:(0.97~1).

[0024] Preferably, the hypoglycemic composition is an oral pharmaceutical composition.

[0025] Beneficial effects:

[0026] This invention uses an ethanol-water solution to extract and enrich components from Huangda tea. Then, by utilizing the dynamic changes in blood glucose and blood components and metabolites in animal models after drug administration, gray-scale correlation analysis (GRA) and partial least squares regression analysis (PLSR) are used to construct a hypoglycemic composition based on Huangda tea. Four active ingredients were obtained, and the ratio of the four active ingredients was adjusted to obtain a hypoglycemic composition with good glucose absorption inhibition effect. Attached Figure Description

[0027] Figure 1 The graph shows the results of partial least squares regression analysis.

[0028] Figure 2 The effect of Huangda tea extract and active substance formulation on cellular glucose consumption. Detailed Implementation

[0029] The following detailed embodiments further illustrate the scheme and effects of the present invention. It should be understood that these embodiments are only used to demonstrate the implementation methods and effects of the present invention and are not intended to limit the scope of the present invention. After reading the content of this invention, those skilled in the art can make various simple modifications or alterations to the present invention without creative effort, and these equivalent / identical transformations also fall within the protection scope of the present invention. In the following embodiments, Huangda tea refers to Huoshan Huangda tea (represented by LYT). The raw material selected for LYT Huoshan Huangda tea processing is one bud and four to five leaves, processed through steps of fixation, rolling, initial drying, yellowing, re-drying, and final firing.

[0030] This invention uses an ethanol-water solution to extract *Tea japonica* (Huangda tea), establishes an animal model, administers the drug to different groups, and monitors the dynamic changes in blood glucose levels and blood components and metabolites in the animal models after drug administration. Correlation analysis using GRA and PLSR is performed to obtain the hypoglycemic active ingredients in *Tea japonica* and its metabolites. A formulation method for a hypoglycemic composition based on *Tea japonica* is constructed, and the ratios of the active ingredients are adjusted to obtain a hypoglycemic composition with good glucose absorption inhibition. The following are specific embodiments of this invention.

[0031] Example 1: Preparation of LYT lyophilized powder

[0032] This embodiment demonstrates a method for preparing LYT lyophilized powder for subsequent animal / in vitro experiments (Examples 2 and 3), and the process is as follows:

[0033] Take an appropriate amount of LYT, pulverize it through an 80-mesh sieve, weigh 15.0 g and place it in a 500 mL Erlenmeyer flask. Add 300 mL of 75% (V / V) ethanol-water at a ratio of 1:20 (W / V), and extract by sonication (360 W, 60℃) for 30 min. Collect the extract and cool it to room temperature. After filtration, concentrate the filtrate under reduced pressure on a rotary evaporator, and then freeze-dry it in a lyophilizer to obtain LYT lyophilized powder (1.0 g of LYT lyophilized powder is equivalent to 3.5 g of LYT). Store it at a low temperature and protected from light.

[0034] The batch-scale extraction method is based on this method and scales up the extraction to meet experimental requirements.

[0035] Example 2 Screening of hypoglycemic active ingredients in Huangda tea and its metabolites

[0036] 1. Establishment and intervention of T2DM rat model

[0037] Healthy male SD rats (180-220g) were acclimatized to a normal diet for one week, then switched to a high-fat diet for six weeks. Blood was collected from the canthal vein to detect biochemical indicators such as triglycerides and total cholesterol. Rats meeting the requirements for a high-fat diet model were selected by fasting overnight for 12 hours. Streptozotocin solution was prepared using citrate buffer at pH 4.2-4.5 and administered intraperitoneally to the high-fat diet model rats at a dose of 35 mg / kg. Blood glucose was measured one week later. Rats with a fasting blood glucose ≥11.1 mmol / L were identified as the T2DM rat model.

[0038] The study included 8 normal control rats and 40 type 2 diabetes mellitus (T2DM) model rats. The T2DM model rats were randomly divided into 5 groups (n=8 per group: model control and different treatment groups). The medium dose of LYT in rats was determined based on the recommended daily tea intake for humans (12g / 60kg) and a conversion factor of 6.3. The rats were administered LYT by gavage once daily for 6 weeks. Specific administration details:

[0039] Normal rats served as the normal control group and were administered 1 mL / (100 g·d) of 0.5% CMC-Na (carboxymethyl cellulose sodium) solution by gavage; the T2DM model control group was administered 1 mL / (100 g·d) of 0.5% CMC-Na solution by gavage; the metformin group was administered 0.20 g / (kg·d) of metformin-0.5% CMC-Na suspension by gavage; and the low, medium, and high dose LYT groups were administered 0.18, 0.35, and 0.70 g / (kg·d) of LYT-0.5% CMC-Na suspension by gavage, respectively. All LYT suspensions were prepared using the lyophilized powder from Example 1, and the dosage was based on LYT, not its extract lyophilized powder.

[0040] 2. Sample collection and its biochemical indicators

[0041] After the last gavage, the patient was kept on a fasting schedule but allowed free access to water for 12 hours. After intraperitoneal anesthesia, the patient underwent dissection to expose the abdominal aorta. Plasma was collected using blood collection tubes containing heparin sodium. The plasma was centrifuged at 3000 rpm for 10 minutes at 4°C and then aliquoted into EP tubes for storage at -80°C.

[0042] Rats’ weight was recorded before the experiment and once a week during the experiment. Rats’ activity, mental state, coat color and survival status were observed daily.

[0043] Ten minutes after the last administration, rats were given glucose solution (2.5 g / kg) by gavage. Blood was collected from the tail vein before the last administration of LYT and at 0 h, 0.25 h, 0.5 h, 1 h, 2 h, and 3 h after administration. Blood glucose levels of T2DM rats were measured using a rapid blood glucose meter to obtain real-time blood glucose values ​​at different time points in each group.

[0044] The results showed that blood glucose levels at all measurement points were significantly lower in the high- and medium-dose LYT groups than in the model group. This suggests that LYT has a significant hypoglycemic effect. The area under the blood glucose-time curve (AUC) in the high- and medium-dose LYT groups was also significantly lower than that in the model group and not significantly different from that in the positive control group (metformin).

[0045] 3. Preparation and processing of plasma from T2DM rats

[0046] The high-dose LYT group was selected as the LYT intervention group, and the T2DM model group was selected as the negative control group for dynamic blood glucose and dynamic blood component (original component) and metabolite detection. Venous blood samples of 0.5 mL were collected before the last administration and at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, and 24 h after administration. After anticoagulation and standing, the samples were centrifuged at 4℃ and 3000 rpm for 10 min. The supernatant was collected, and 100 μL of cold methanol was added to every 200 μL to precipitate proteins. After vortexing and mixing, the samples were centrifuged at 4℃ and 12500 rpm for 10 min. The supernatant was collected, dried under nitrogen, and the residue was reconstituted with 100 μL of cold methanol. After vortexing and centrifugation, the supernatant was placed in a vial with a glass inner tube for UPLC-Q / TOF-MS analysis. Plasma samples from the T2DM model group were processed in the same way as a negative control.

[0047] 4.UPLC-Q / TOF-MS analysis

[0048] UPLC conditions: A Waters ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) was used, with a column temperature of 35 °C. The mobile phase was 0.1% formic acid in water (A) and acetonitrile (B), and the flow rate was 0.2 mL / min. -1The injection volume was 2 μL. The gradient elution program was as follows: 0–5 min (95% A), 5–10 min (95% → 90% A), 10–20 min (90% → 84% A), 20–32 min (84% → 80% A), 32–37 min (80% → 65% A), 37–42 min (65% → 45% A), 42–45 min (45% → 30% A), 45–50 min (30% → 10% A), 50–53 min (10% A), 53–54 min (10% → 95% A), and 54–55 min (95% A).

[0049] Q / TOF-MS conditions: Electrospray ionization (ESI) source with dual-mode detection (positive and negative ions), ion source temperature 120℃, scan range m / z 50-1200 Da, capillary voltage 3.0 kV (positive mode) / 2.5 kV (negative mode), cone voltage 40.0 kV, solvation temperature 350℃, solvent gas flow rate 600 L·h -1 Mass axis correction uses leucine-enkephalin (positive mode [M+H]). + =556.2771, Negative Mode [MH] - =554.2615), MS E The collision energy ranges from 6 V for low energy to 50-60 V for high energy; data acquisition and processing are performed using MassLynx 4.1 software (Waters Corporation).

[0050] 5. Identification of LYT precursor components and metabolites in rat plasma

[0051] The first step involved constructing a self-built LYT library based on the research group's previous analysis of the chemical composition of LYT. This library was developed using the chromatographic behavior (retention time), mass spectrometry characteristics (secondary fragment ion information, characteristic fragment ions, quasi-molecular ions), and chemical composition information (molecular formula, precise molecular weight, structural formula, etc.) of the 87 identified compounds. High-resolution mass spectrometry data were analyzed using a MassLynx 4.1 workstation. Ion currents with m / z values ​​matching (error ±10 ppm) and consistent retention times in blood samples after LYT administration were extracted. Combined with fragment ion characteristics at high collision energies (50–60 V), the original components absorbed into the bloodstream by LYT were identified, and interference from extracted ion chromatograms (EIC) in the blank control group was eliminated, resulting in total ion chromatograms of LYT plasma and blank plasma samples.

[0052] The second step involves integrating the identified original components (including molecular formulas and structural .mol ​​files) in the blood after LYT administration and the components with larger response signals in the LYT samples, reconstructing the database, and importing it along with the raw data collected by the instrument into the UNIFI platform.

[0053] The third step involves setting the metabolite identification parameters in UNIFI: mass-to-charge ratio error ±10 ppm; and adduct ions for positive / negative ion modes as [M+H]. + / [MH] - The metabolic reaction types cover phase I (oxidation, reduction, demethylation, etc.) and phase II (glucuronidation, sulfation, etc.); at the same time, the detection thresholds are set as 2D peak area ≥200 and 3D peak intensity ≥500 (low energy) or ≥100 (high energy).

[0054] Finally, using UNIFI automatic matching combined with MassLynx fragmentation data, the retention time, precise mass, fragmentation pattern, and structural rationality of candidate metabolites were manually verified. Overlapping signals from the blank group were removed, ultimately confirming the metabolites of LYT in rat plasma. The detected LYT components and metabolites are shown in Tables 1 and 2.

[0055] Table 1. Information on LYT components and metabolites entering the bloodstream.

[0056] Table 2 Information on LYT components and metabolites entering the bloodstream.

[0057] Overall metabolic type: In rat plasma, the metabolism of LYT mainly involves phase I reactions (oxidation, reduction, demethylation, deglycosylation, etc.) and phase II reactions (glucuronidation, acetylation, methylation, sulfation, etc.). Most metabolites are generated by the synergistic effect of phase I and phase II reactions.

[0058] 6. Establishment of the "spectrum-effect" relationship

[0059] Based on the identified migrating components analyzed by UPLC-ESI-QTOF / MS, the peak areas of each component at different blood collection time points were extracted as "spectrums".

[0060] Blood glucose levels at different time points were measured in model rats after gavage administration of glucose. i The different time points corresponding to the blood glucose collection are t i The area under the blood glucose-time curve was divided into multiple trapezoids using the trapezoidal method, and the blood glucose AUC of each group was calculated as A. i .

[0061]

[0062] (Formula 1).

[0063] Based on the above method, A values ​​for the T2DM group and the LYT-H group at different time points were calculated respectively. iWith two groups of A at the same time i The difference is used as the "effect" in the spectral effect relationship.

[0064] Based on the constructed "spectrum" and "efficacy" data sets, a "spectrum"-"efficacy" correlation model was established through GRA and PLSR analyses. When a compound has a grey relational degree ≥ 0.8, a VIP value highly close to 1, and a negative correlation with the efficacy index, it can be included in the candidate category of active substances. PLSR results are as follows: Figure 1 As shown.

[0065] Four active substances were ultimately obtained that met the criteria. All of these are the original components of LYT, not its metabolites: (2S,3R)-gallocatechin (see M7 in Table 2), (2S,3R)-Epigallocatechin (see M9 in Table 2), 3-p-Coumaroylquinic acid (see M8 in Table 2), and Isoquercitrin (see M14 in Table 2). These four substances will be used as active ingredients in the formulation of a hypoglycemic composition.

[0066] Example 3: Investigation of the formulation ratio of the hypoglycemic composition

[0067] 1. Determination of active substance content

[0068] This method is used to detect the content of corresponding active ingredients in Huangda tea extract, thereby obtaining the quality information of the active ingredients as a benchmark for adjusting the formulation of hypoglycemic compositions.

[0069] UPLC conditions: The column was a Waters BEH C18 (2.1 mm × 100 mm, 1.7 μm); the mobile phase was acetonitrile (A) – 0.1% formic acid aqueous solution (B) gradient elution, with the elution program as follows: 0–5 min, 5–14% A; 5–9 min, 14–20% A; 9–12 min, 20% A; 12–15 min, 20–5% A; 15–17 min, 5% A. The flow rate was 0.2 mL / min. -1 The detection wavelength was 280 nm; the column temperature was 30℃; and the injection volume was 2 μL.

[0070] Preparation of mixed reference solutions: Accurately weigh appropriate amounts of reference standards for (2S,3R)-gallocatechin, (2S,3R)-Epigallocatechin, 3-p-Coumaroylquinic acid, and isoquercitrin, and place them separately in 10 mL volumetric flasks. Dissolve and dilute to the mark with methanol, and shake well to obtain mixed standard solutions of appropriate mass concentrations. Prepare five series of mixed reference standard solutions with concentrations of 1, 5, 10, 50, and 100 μg / mL by diluting an appropriate amount of the mixed standard solution with methanol. Perform chromatographic analysis under the above conditions, and perform linear regression of peak area (Y) against mass concentration (X, μg / mL).

[0071] Accurately weigh 1g of LYT lyophilized powder, add 10mL of 70% methanol-water (V / V), sonicate, centrifuge an appropriate amount of solution (3000rpm, 10min), filter the supernatant through a 0.22μm filter membrane, and collect it into a vial to obtain the LYT sample solution. Inject the sample according to the above injection method and perform quantitative analysis using the external standard method.

[0072] 2. LYT and its active ingredients intervene in insulin resistance (IR) in HepG2 cells.

[0073] Single-cell suspensions were prepared from HepG2 cells in the logarithmic growth phase. After counting, the cell density was adjusted to 2 × 10⁶ cells using complete culture medium. 4 Cells / mL were seeded at 100 μL per well in 96-well plates and cultured at 37°C in a 5% CO2 incubator until cell adhesion occurred. The original culture medium was discarded, and serum-free culture medium was added to starve the cells for 12 h. After washing twice with PBS buffer, the cells were divided into groups for further treatment.

[0074] Normal control group: serum-free culture medium was added;

[0075] Model group: serum-free medium containing 18 mmol / L glucosamine (GlcN) was added, and the medium was replaced with serum-free medium 24 h after intervention;

[0076] LYT intervention group: serum-free medium containing 18 mmol / L GlcN was added. After 24 h of intervention, serum-free medium containing LYT was added (final concentrations were 500 μg / mL, 200 μg / mL, and 100 μg / mL, respectively, based on LYT rather than its extract lyophilized powder).

[0077] All groups were cultured for another 24 hours. The amount of glucose remaining in the supernatant was measured using a glucose assay kit, and the glucose consumption was calculated. The optimal intervention concentration of LYT was determined by comparing the differences between the model group and each intervention group (see [link to intervention group]). Figure 2A). Figure 2 A showed that in HepG2 cells with insulin resistance, treatment with 200 μg / mL LYT most significantly increased glucose consumption, indicating that this concentration had the best effect on improving insulin resistance; 100 μg / mL had the second best effect; while the 500 μg / mL group showed no significant difference. This is usually because excessively high concentrations produce cytotoxic or inhibitory effects, which interfere with the normal glucose uptake function of cells, leading to the disappearance of the improvement effect.

[0078] 3. The proportions of the hypoglycemic composition should be carefully selected.

[0079] Based on the optimal LYT concentration determined above, the mass ratio of the four active ingredients (calculated by concentration) was used as the variable of investigation, with three concentration levels for each ingredient (see Table 3). The L9(3) method was employed. 4 The experiments were arranged using an orthogonal array (Table 4).

[0080] Table 3 Factor Level Table

[0081]

[0082] Table 4 Orthogonal Experiment Table

[0083]

[0084] HepG2 cells were starved and then grouped as follows:

[0085] Model group: Serum-free medium containing 18 mmol / L GlcN was added, and the medium was replaced with serum-free medium after 24 h;

[0086] Active substance combination intervention group: Serum-free medium containing 18 mmol / L GlcN was added, and after 24 h, it was replaced with serum-free medium containing the corresponding orthogonal combination of active substances. After another 24 h of culture, the glucose consumption of cells was used as the evaluation index. The differences between the model group and each active substance combination group were compared to determine the optimal combination of active substances for regulating insulin resistance in HepG2 cells (see...). Figure 2 B).

[0087] The results showed that among all the active substance combinations, the fifth group, consisting of gallocatechin 0.44 μg / mL, epigallocatechin 0.52 μg / mL, 3-coumaroylquinic acid 0.44 μg / mL, and isoquercitrin 0.43 μg / mL, exhibited the most significant regulatory effect on insulin resistance in HepG2 cells. This effect was not only significantly better than the model group but also demonstrated a unique and significant advantage among all the combination groups. The synergistic regulatory effect of the four active substances depended on the rational matching of the concentrations of each component, rather than a simple additive effect. The specific low-dose near-concentration combination screened in this study was determined through systematic exploration among various potential ratios and could not be directly obtained through conventional concentration trials or empirical deduction. Therefore, the fifth group can be identified as the optimal combination of active substances for regulating insulin resistance (IR) in HepG2 cells. When gallocatechin, epigallocatechin, 3-p-coumarylquinic acid and isoquercitrin are used instead of LYT, the approximate mass ratio of the four components is 1:1.18:1:0.98.

Claims

1. A method of formulating a hypoglycemic composition based on Camellia sinensis, characterized in that Comprising the following steps: S1 Preparation of Huangda tea extract: Huangda tea is extracted with 70%-75% ethanol aqueous solution, the extract is concentrated and solvent is removed to prepare Huangda tea extract; S2 Selection of ingredients for prescription: establish a type 2 diabetes animal model, give Huangda tea extract by gavage, respectively determine the area under the blood glucose-time curve Ai of the model animals and the drug administration animals and the concentration or chromatographic peak area of the original components and metabolites of Huangda tea in the blood samples; use grey correlation analysis method and partial least squares regression analysis method to analyze the correlation degree of the Ai difference of the model animals and the drug administration animals and the concentration or chromatographic peak area of the original components and metabolites of Huangda tea in the blood samples, take the original components and / or metabolites of the drugs with grey correlation degree≥0.8, VIP value close to 1 and negative correlation with animal blood glucose value as the ingredients for prescription; S3 Prescription of hypoglycemic composition: take the ingredients for prescription of step S2 as active ingredients and combine them to prepare a hypoglycemic composition.

2. A method of formulating a hypoglycemic composition based on yellow tea as claimed in claim 1, wherein, In step S2, UPLC-Q-TOF / MS is used to determine the concentration or chromatographic peak area of the original components and metabolites of Huangda tea in the blood samples.

3. A method of formulating a hypoglycemic composition based on yellow tea as claimed in claim 2, wherein, The detection conditions of the UPLC-Q-TOF / MS include: The UPLC conditions: use C18 chromatographic column as the stationary phase, the chromatographic column specifications are: 100 mm x 2.1 mm, 1.7 μm; the mobile phase A is 0.1% formic acid water, the mobile phase B is acetonitrile, the flow rate is 0.2 mL / min, and the gradient elution program is: 0-5 min (95% A), 5-10 min (95%→90% A), 10-20 min (90%→84% A), 20-32 min (84%→80% A), 32-37 min (80%→65% A), 37-42 min (65%→45% A), 42-45 min (45%→30% A), 45-50 min (30%→10% A), 50-53 min (10% A), 53-54 min (10%→95% A), 54-55 min (95% A).

4. A method of formulating a hypoglycemic composition based on yellow tea as claimed in claim 2, wherein, The detection conditions of the UPLC-Q-TOF / MS include: Q / TOF-MS conditions: positive and negative ion dual mode detection was used by electrospray ionization source, ion source temperature 120℃, scanning range m / z 50-1200 Da, capillary voltage 3.0 kV (positive mode) / 2.5 kV (negative mode), cone voltage 40.0 kV, solventation temperature 350℃, solvent gas flow rate 600 L·h -1 ; mass axis correction used leucine-enkephalin, positive mode [M+H] + =556.2771, negative mode [M-H] - =554.2615, MSE collision energy was low energy 6V, high energy 50~60V.

5. A method of formulating a hypoglycemic composition based on yellow tea as claimed in claim 1, wherein, In step S2, the type 2 diabetes animal model is a rodent model.

6. The method of claim 1, wherein the composition is a tea composition. In step S2, the method for calculating the area Ai under the blood glucose-time curve is as follows: the blood glucose value at different time points after modeling is G i The different time points for collecting blood glucose are t i The area under the blood glucose-time curve is divided into multiple trapezoids by using the trapezoidal method, and the area under the blood glucose-time curve is calculated as Ai. 。 7. A method of formulating a hypoglycemic composition based on yellow tea as claimed in claim 1, wherein, In step S3, the mass ratio of active ingredients in the hypoglycemic composition is the ratio of the content of the corresponding active ingredients in Huangda tea extract.

8. A hypoglycemic composition obtained by the method of claim 1-6, wherein the hypoglycemic composition is characterized in that, The active ingredients of the hypoglycemic composition are: (2S, 3R)-gallocatechin, (2S, 3R)-Epigallocatechin, 3-p-Coumaroylquinic acid and Isoquercitrin.

9. The antihyperglycemic composition of claim 8, wherein, The mass ratio of (2S, 3R)-gallocatechin, (2S, 3R)-Epigallocatechin, 3-p-Coumaroylquinic acid and Isoquercitrin in the hypoglycemic composition is 1:(1.17-1.2):1:(0.97-1).

10. The antihyperglycemic composition of claim 8, wherein The hypoglycemic composition is an oral pharmaceutical composition.

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