Chrysanthemum extract, preparation method and application

Abstract

The invention belongs to the technical field of chrysanthemum extracts, and particularly relates to a chrysanthemum extract, a preparation method and application. The chrysanthemum extract contains chrysanthemum flavonoid glycoside, and the chrysanthemum flavonoid glycoside comprises the following main components: luteolin-7-O-rutinoside of which the molecular formula is C27H30O15, luteolin-7-O-glucoside of which the molecular formula is C21H20O11, luteolin-7-O-glucuronide of which the molecular formula is C21H18O12, luteolin-7-O-rutinoside of which the molecular formula is C27H30O15, luteolin-7-O-glucuronide of which the molecular formula is C21H20O11, luteolin-7-O-rutinoside of which the molecular formula is The molecular formula of the luteolin-7-O-6 ''-malonyl glucoside is C24H22O14, the molecular formula of the luteolin-7-O-2''-acetyl glucuronide is C23H20O13, and the molecular formula of the luteolin-7-O-6 ''-acetyl glucuronide is C23H22O12. The invention further discloses a preparation method of the luteolin-7-O-6''-malonyl glucoside. The chrysanthemum flavonoid glycoside serving as a raw material is applied to preparation of a blood fat reducing medicine or a medicine for enhancing the antioxidant capacity of an organism. The product is feather-shaped and layered, the whole product is light, thin, easy to absorb water and extremely easy to dissolve in water, and an aqueous solution is bright yellow; wherein the luteoloside is rich in luteoloside; the health-care tea has the functions of reducing blood fat and enhancing the antioxidant capacity of the body.

Description

Technical Field

[0001] The invention belongs to the technical field of chrysanthemum extracts, and particularly relates to a chrysanthemum extract, a preparation method and an application thereof. Background Art

[0002] Chrysanthemum, scientific name Dendranthema morifolium, Latin name Flos Chrysanthemi, is a perennial herbaceous plant of the Asteraceae family that can be used as both medicine and food. Its petals are tongue-shaped or tubular. The dried capitulum of chrysanthemum is a commonly used Chinese medicine in China. It has the effects of dispelling wind, clearing heat, improving eyesight, and detoxifying. It is mainly used to treat headaches, dizziness, red eyes, chest heat, furuncles, swelling and poison, etc. It is also often used as a health food for lowering blood pressure. Studies have found that the chemical composition of chrysanthemum is relatively complex, among which flavonoids, triterpenoids and volatile oils are its main active ingredients. According to literature reports, chrysanthemum extracts have multiple pharmacological activities such as antibacterial, antiviral, anti-inflammatory and immune, cardiovascular system protection, anti-tumor, liver protection and neuroprotection (Chen Chuanqian et al., Research Progress on the Pharmacological Effects of Wild Chrysanthemum Extracts). However, there are no related research reports on chrysanthemum and its extracts in the prevention and treatment of fatty liver. Summary of the invention

[0003] In view of the deficiencies in the prior art, the present invention provides a chrysanthemum extract, a preparation method and an application thereof.

[0004] The purpose of the present invention can be achieved through the following technical solutions:

[0005] A chrysanthemum extract, the chrysanthemum extract contains chrysanthemum flavonoids, and the main components of the chrysanthemum flavonoids include:

[0006] Luteolin-7-O-rutinoside, whose molecular formula is C 27 H 30 O 15 , the structural formula is:

[0007]

[0008] Luteolin-7-O-glucoside, whose molecular formula is C 21 H 20 O 11 , the structural formula is:

[0009]

[0010] Luteolin-7-O-glucuronide, whose molecular formula is C 21 H 18 O 12 , the structural formula is:

[0011]

[0012] Luteolin-7-O-6"-malonyl glucoside, whose molecular formula is C 24 H 22 O 14 , the structural formula is:

[0013]

[0014] Luteolin-7-O-2"-acetyl glucuronide, whose molecular formula is C 23 H 20 O 13 , the structural formula is:

[0015]

[0016] Luteolin-7-O-6"-acetyl glucuronide, whose molecular formula is C 23 H 22 O 12 , the structural formula is:

[0017]

[0018] A method for preparing a chrysanthemum extract, the method comprising the following steps:

[0019] S1, removing impurities from the picked chrysanthemums to retain the complete inflorescence, and then drying them to obtain dried chrysanthemums;

[0020] S2, crushing and grinding the dried chrysanthemum, and then sieving to obtain chrysanthemum powder;

[0021] S3, using an ethanol solution to fully soak the chrysanthemum powder, and then subjecting the chrysanthemum powder to ultrasonic oscillation to obtain a chrysanthemum powder soaking solution;

[0022] S4, extracting the chrysanthemum powder soaking solution in a water bath to obtain an extract;

[0023] S5, filtering the extract to remove impurities, and then concentrating the extract to 1% of the volume of the original solution to obtain a concentrated solution;

[0024] S6. freeze-drying the concentrated solution to obtain a crude chrysanthemum extract;

[0025] S7. Dissolve the crude chrysanthemum extract in water to prepare a chrysanthemum solution, adjust the pH of the chrysanthemum solution to 2.5, and then flow into a macroporous adsorption resin at a flow rate of 1.00 mL / min; use an ethanol solution to analyze the macroporous adsorption resin adsorbed with the chrysanthemum solution; and obtain a chrysanthemum extract after concentrating and freeze-drying the effluent.

[0026] Furthermore, in step S1, the drying method adopts a stepwise low-temperature drying method, and the temperature is set to 25°C - 35°C - 45°C - 35°C - 25°C.

[0027] Furthermore, in step S2, a sieve with a mesh size of 200 to 250 is used for screening.

[0028] Furthermore, in step S4, the material-liquid ratio of the chrysanthemum powder and the clean water is 1:40; the water bath temperature is 85° C. to 100° C., and the water bath time is 45 to 60 min.

[0029] Furthermore, the concentration of the ethanol solution used in steps S3 and S7 is 70%.

[0030] Furthermore, the concentration of the chrysanthemum solution in step S7 is 3.60 mg / mL, and the volume of the chrysanthemum solution is twice the volume of the macroporous adsorption resin; when using ethanol solution for analysis, the volume of the ethanol solution is twice the volume of the macroporous adsorption resin.

[0031] The above-mentioned chrysanthemum extract is used as a raw material in the preparation of lipid-lowering drugs.

[0032] The above-mentioned chrysanthemum extract is used as a raw material in the preparation of a drug for enhancing the body's antioxidant capacity.

[0033] The beneficial effects of the present invention are as follows: the product is feather-shaped, layered, thin and light as a whole, easy to absorb water, extremely soluble in water, and the aqueous solution is bright yellow; the product is rich in luteolin; it has the effects of lowering blood lipids and enhancing the body's antioxidant capacity. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0035] Figure 1 is a mass spectrometry analysis diagram of luteolin-7-O-rutinoside in the present invention;

[0036] Figure 2 is a mass spectrometry analysis diagram of luteolin-7-O-glucoside in the present invention;

[0037] Figure 3 is a mass spectrometry analysis diagram of luteolin-7-O-glucuronide in the present invention;

[0038] Figure 4 is a mass spectrometry analysis diagram of luteolin-7-O-6"-malonyl glucoside in the present invention;

[0039] Figure 5 The mass spectrum analysis diagram of luteolin-7-O-2"-acetyl glucuronide in the present invention;

[0040] Figure 6 The mass spectrum analysis diagram of luteolin-7-O-6"-acetyl glucuronide in the present invention;

[0041] Figure 7 This is a full-wavelength ultraviolet scanning diagram of the chrysanthemum extract of the present invention;

[0042] Figure 8 This is the infrared spectrum of the chrysanthemum extract in the present invention;

[0043] Fig. 9 This is a fluorescence emission spectrum of the chrysanthemum extract of the present invention;

[0044] Fig.10 These are actual pictures of the chrysanthemum extract of the present invention, wherein the left picture is a crude chrysanthemum extract; the middle picture is a purified chrysanthemum extract; and the right picture is a solution of the chrysanthemum extract dissolved in water. DETAILED DESCRIPTION

[0045] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0046] A method for preparing chrysanthemum flavonoid glycosides, the preparation method comprising the following steps:

[0047] S1. Remove impurities from the picked chrysanthemums to retain the complete inflorescence, and then dry them to obtain dried chrysanthemums; the drying method adopts a step-by-step low-temperature drying method, and the temperature is set to 25°C-35°C-45°C-35°C-25°C. This method can retain the flower shape and the effective ingredients in the chrysanthemums to the maximum extent;

[0048] S2, using a universal grinder to preliminarily grind the chrysanthemum, then further refine it using a grinding mill, and then use a 200-250 mesh sieve to grade and screen to obtain chrysanthemum powder;

[0049] S3, using a 70% ethanol solution to fully soak the chrysanthemum powder, and then using an ultrasonic device to oscillate for 30 minutes to obtain the soaked chrysanthemum powder;

[0050] S4. Place the soaked chrysanthemum powder in clean water for water bath extraction to obtain an extract; the solid-liquid ratio of the soaked chrysanthemum powder to clean water is 1:40; the water bath temperature is 91° C., and the water bath time is 51 min.

[0051] S5, filtering the extract to remove impurities, and then concentrating the extract to 1% of the volume of the original solution to obtain a concentrated solution;

[0052] S6. The extract is first cryogenically frozen for 24 hours, and then vacuum freeze-dried for 48 hours to obtain a crude chrysanthemum extract.

[0053] S7. Dissolve the crude chrysanthemum extract in water to prepare a chrysanthemum solution with a concentration of 3.60 mg / mL, and adjust the pH of the chrysanthemum solution to 2.5, and then flow into the macroporous adsorption resin at a rate of 1.00 mL / min, wherein the volume of the chrysanthemum solution is twice the volume of the macroporous adsorption resin; then use a 70% ethanol solution to analyze the macroporous adsorption resin adsorbed with the chrysanthemum solution; the volume of the ethanol solution is twice the volume of the macroporous adsorption resin; the effluent is concentrated and freeze-dried to obtain chrysanthemum flavonoids.

[0054] The components of the chrysanthemum flavonoid glycosides were analyzed by mass spectrometry; the main components obtained by analysis are as follows:

[0055] 1. Luteolin-7-O-rutinoside

[0056] The peak time of mass spectrometry analysis was 11.18, [MH]- was m / z 593.1508, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 27 H 30 O 15 According to the secondary mass spectrometry, the parent nucleus of the compound is 285. According to literature research, the parent nucleus may be luteolin. In addition, the fragment ion m / z 285 [M-rutinoside] was observed, indicating that there is a rutinose in the structure. Combined with the database search, it is speculated that component 1 is luteolin-7-O-rutinoside; its structural formula is:

[0057]

[0058] 2. Luteolin-7-O-glucoside

[0059] The peak time of mass spectrometry analysis was 11.72, [MH]- was m / z 447.0926, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 21 H 20 O 11 According to the secondary mass spectrometry, the parent nucleus of the compound is 285, which is luteolin. In addition, the fragment ion m / z 285[M-Glc]- was observed, indicating that there is a glucose in the structure. Combined with the database search, it is speculated that component 2 is luteolin-7-O-glucoside; its structural formula is:

[0060]

[0061] 3. Luteolin-7-O-glucuronide

[0062] The peak time of mass spectrometry analysis was 11.72, [MH]- was m / z 461.0719, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 21 H 18 O 12 According to the secondary mass spectrometry, the parent nucleus of the compound is 285, which is luteolin. In addition, the fragment ion m / z 285[M-Glu]- was observed, indicating that there is a glucuronic acid in the structure. Combined with the database search and speculation, component 3 is luteolin-7-O-glucuronide; its structural formula is:

[0063]

[0064] 4. Luteolin-7-O-6”-malonyl glucoside

[0065] The peak time of mass spectrometry analysis was 13.24, [MH]- was m / z 533.0930, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 24 H 22 O 14 According to the secondary mass spectrometry, the parent nucleus of the compound is 285, which is luteolin. The compound easily loses CO2, indicating that there may be a malonic acid monoacyl structure in the structure. Combined with the database search, it is speculated that component 4 is luteolin-7-O-6"-malonyl glucoside; its structural formula is:

[0066]

[0067] 5. Luteolin-7-O-2”-acetyl glucuronide

[0068] The peak time of mass spectrometry analysis was 13.99, [MH]- was m / z 503.0823, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 23 H 20 O 13 According to the secondary mass spectrometry, the parent nucleus of the compound is 285, which is luteolin. In addition, the fragment ion m / z 285 [M-Glu-Ac] was observed, indicating that the structure contains 1 glucuronic acid and 1 acetylated group. Combined with the database search, it is speculated that component 5 is luteolin-7-O-2"-acetyl glucuronide; its structural formula is:

[0069]

[0070] 6. Luteolin-7-O-6”-acetyl glucuronide

[0071] The peak time of mass spectrometry analysis was 14.94, [MH]- was m / z 489.1029, and the molecular formula fitted based on the high-resolution mass spectrometry results was C 23 H 22 O 12 According to the secondary mass spectrometry, the parent nucleus of the compound is 285, which is luteolin. In addition, the fragment ion m / z 285 [M-Glc-Ac] was observed, indicating that the structure contains 1 glucose and 1 acetylated group. Combined with the database search, it is speculated that component 6 is luteolin-7-O-6"-acetyl glucuronide, and its structural formula is:

[0072]

[0073] The chrysanthemum extract was subjected to spectral analysis, and the analysis results were as follows: Figure 7 to Figure 9 shown.

[0074] It can be seen from the figure that flavonoids have two main absorption bands in the ultraviolet range, namely absorption band I and absorption band II. The maximum absorption peak of flavonoids is mainly concentrated in absorption band II ( Figure 7 The peaks appearing at 1750-1600cm-1 are mainly the breathing vibration of the carbon skeleton ring of the benzene ring and the stretching vibrations of C=N, C=O, C=C, etc. The peaks belong to flavonoids. The appearance of two peaks in this range indicates that the flavonoid content is richer, and the highest absorption peak appears at 250-280nm ( Figure 8 ). Flavonoids have a multi-ring structure and therefore have strong fluorescence properties when excited ( Fig. 9 ).

[0075] The efficacy of chrysanthemum extract was experimentally analyzed.

[0076] Example 1: Using chrysanthemum extract in animal experiments

[0077] 1.1 Experimental Animals

[0078] Sixty male SD rats (SPF grade), weighing 200±9g, were purchased from the Experimental Animal Base of Nanjing Medical University, with the license number SYXK(Su)2016-0015. The experimental animals were raised in the SPF animal experimental center of the Dingjiaqiao campus of Southeast University. During the entire experiment, the indoor temperature was controlled at 22℃~24℃, the humidity was maintained at 40%~60%, the day and night cycle was 12h, and they were free to drink water and eat. The animals were used in the experiment after 1 week of adaptive feeding. The experiment was approved by the Animal Ethics Committee of Southeast University, and all procedures involving animals were in accordance with the operating guidelines for laboratory animal care of Southeast University.

[0079] 1.2 Experimental Materials

[0080] Chrysanthemum flavonoids: Chrysanthemum flavonoids were extracted according to the method of "Part 1". Animal feed: Low-fat feed (D12450B) and high-fat feed (D12492) were purchased from Jiangsu Collaborative Biomedicine Co., Ltd. The fat energy supply ratio of low-fat feed (D12450B) and high-fat feed (D12492) was 10% and 60%, respectively. The nutrient content and energy supply of the two feeds are shown in Table 1. D12492 high-fat feed was used to model the high-fat model of rats, which is closer to the characteristics of hyperlipidemia.

[0081] Table 1 Nutrients and energy supply of D12450B feed and D12492 feed

[0082]

[0083] 1.3 Modeling and grouping

[0084] After one week of adaptive feeding, 60 male SD rats were randomly divided into a normal control group (n=10) and a high-fat model group (n=50). The normal group and the high-fat model group were given D12450B feed and D12492 high-fat feed, respectively. The modeling period was 4 weeks. After 4 weeks, blood was collected by tail cutting in the morning to determine the levels of TC and TG in serum. If the TG and TC levels in the model group were significantly higher than those in the normal control group (p<0.05), the model was judged to be successful.

[0085] Antioxidant experiments and content detection have proved that luteolin is the main active ingredient of chrysanthemum flavonoids. In order to further study the hypolipidemic effect and mechanism of chrysanthemum flavonoids and its important active ingredient luteolin, after successful modeling, the model rats were randomly divided into five groups: high-fat model group, chrysanthemum flavonoids group, luteolin group, and simvastatin group (positive control group), with 10 rats in each group. The normal control group was fed with D12450B feed, and the other groups were fed with D12492 high-fat feed. The intervention period was 6 weeks, and each group of animals had free access to water and food. The experimental design and intervention dose are shown in Table 2. The intragastric dose of chrysanthemum flavonoids, luteolin, luteolin, and simvastatin in each group was carried out according to Table 2. Specifically, the intragastric solution of the corresponding concentration was prepared according to the intragastric volume of 10 ml / kg, and the solvent was 5‰ sodium carboxymethyl cellulose (CMC) solution.

[0086] Table 2 Experimental groups and interventions

[0087]

[0088] 1.3. Determination of general indicators such as body weight and organ coefficients

[0089] During the intervention period, rats were weighed once every Monday morning and recorded to observe changes in body weight over the intervention period. At the end of the intervention, rats were fasted for 12 hours, and blood was collected from the femoral artery after weighing and anesthesia, and then centrifuged and separated. The serum was stored at -20°C, and the upper serum was used to detect relevant biochemical indicators. Afterwards, the rats were killed by cervical dislocation, and the changes in fat and liver and other organs in the rats were observed by dissection. The liver and total visceral fat were then quickly separated and weighed separately. Based on the wet weight of the liver and the total fat weight, the liver coefficient and body fat rate were estimated in combination with the body weight of each group of SD rats. The liver coefficient = wet weight of the liver / body weight × 100%, and the body fat rate = total fat weight / body weight × 100%. In addition, serum, feces, liver and peritesticular fat were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator for later use.

[0090] 1.4. Determination of four blood lipids, apolipoprotein A1 and apolipoprotein B

[0091] Take 2.5 μL serum sample, and measure four blood lipids and apolipoprotein according to the kit instructions. TC and TG levels in serum were measured by enzyme method, HDL-C and LDL-C levels in serum were measured by direct method, Apo-A1 and Apo-B levels in serum were measured by enzyme linked immunosorbent assay (ELISA), and Apo-B / Apo-A1 was calculated.

[0092] 1.5. Pathological section analysis of liver and peritesticular fat tissue

[0093] The liver tissue and peritesticular adipose tissue fixed with 10% formaldehyde solution were dehydrated, paraffin-embedded, sliced ​​(4 μm thick) and stained with HE. The pathological changes of liver and adipose tissue were observed using an Olympus BX41 microscope. The degree of fatty degeneration of the liver was quantitatively analyzed by using Imge-pro Plus 6.0 image analysis software according to the degree of fatty degeneration score: 0 points indicated normal, 1 point indicated that the degree of fatty degeneration of the liver was less than 5%, 2 points indicated that the degree of fatty degeneration of the liver was 5% to 30%, 4 points indicated that the degree of fatty degeneration of the liver was 50% to 75%, and 5 points indicated that the degree of fatty degeneration of the liver was greater than 75%. The eyepiece was set to 10* and the objective lens to 20*. The number of fat cells in an area of ​​25 mm2 (cells / 25 mm2 / 200×) was measured using a "micrometer for microscope" under a 200x light microscope to evaluate the size of fat cells in the pathological sections of adipose tissue.

[0094] 1.5. Determination of liver antioxidant system-related enzymes and malondialdehyde

[0095] According to the instructions of the kit, the contents of malondialdehyde (MDA), glutathione catalase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) in the liver were determined by TBA method, colorimetry, visible light method and hydroxylamine method respectively.

[0096] 1.6 Experimental Results

[0097] Table 3 Liver wet weight, total fat, body weight liver coefficient and body fat percentage of SD rats in each group

[0098]

[0099] Note: a P < 0.05, compared with the normal control group; b P<0.05, compared with the high-fat model control group; c P<0.05, compared with the chrysanthemum flavonoids group; n=10.

[0100] As shown in Table 3, the liver wet weight, total fat, liver coefficient and body fat rate in the high-fat model group were significantly higher than those in the normal control group (P<0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the liver wet weight, total fat, liver coefficient and body fat rate were significantly decreased (P<0.05), and there was no statistical difference with the normal control group (P>0.05). Compared with the chrysanthemum flavonoids group, there was no statistical difference in liver wet weight, total fat, liver coefficient and body fat rate in the luteolin group and simvastatin group (P>0.05).

[0101] As shown in Table 3, the body weight of the high-fat model group was significantly higher than that of the normal control group (P<0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the body weight of the chrysanthemum flavonoids group showed a downward trend but no statistical difference (P>0.05), and both were significantly increased compared with the normal control group (P<0.05). The body weight of the luteolin group and the simvastatin group decreased significantly (P<0.05), and both were not statistically different from the normal control group (P>0.05). Compared with the chrysanthemum flavonoids group, the body weight of the luteolin group and the simvastatin group was not statistically different (P>0.05).

[0102] Table 4 Changes of four blood lipids in SD rats in each group

[0103]

[0104] Note: a P < 0.05, compared with the normal control group; b P < 0.05, compared with the model control group;c P<0.05, compared with the chrysanthemum flavonoids group; n=10.

[0105] As shown in Table 4, the levels of TC, TG and LDL-C in the serum of the hyperlipidemia model group were significantly higher than those of the normal control group (P<0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the levels of TC, TG and LDL-C in the serum were significantly decreased (P<0.05), and there was no statistical difference between them and the normal group (P>0.05). Compared with the chrysanthemum flavonoids group, there was no statistical difference in the levels of TC, TG and LDL-C in the serum of the simvastatin group and the luteolin group (P>0.05).

[0106] The HDL-C content in the serum of the hyperlipidemia model group was significantly lower than that in the normal control group (P<0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the HDL-C content in the serum showed an upward trend but no statistical difference. Compared with the normal control group, the HDL-C content in the serum of the chrysanthemum flavonoids group was significantly decreased (P<0.05), and there was no statistical difference in the HDL-C content in the serum of the luteolin group and the simvastatin group (P>0.05). Compared with the chrysanthemum flavonoids group, there was no statistical difference in the HDL-C content in the serum of the luteolin group and the simvastatin group (P>0.05).

[0107] Table 5 Changes of apolipoprotein Apo-A1, Apo-B, Apo-B / Apo-A1 in rats of each group

[0108]

[0109] Note: a P < 0.05, compared with the normal control group; b P<0.05, compared with the high-fat model control group; c P<0.05, compared with the chrysanthemum flavonoids group; n=10.

[0110] As shown in Table 5, the Apo-A1 content in the serum of the high-fat model group was significantly lower than that in the normal control group (P<0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the Apo-A1 content in the serum of the chrysanthemum flavonoids group and the luteolin group increased significantly (P<0.05), and there was no statistical difference in the Apo-A1 content in the serum of the simvastatin group (P>0.05), and there was no statistical difference between the two groups and the normal control group (P>0.05). Compared with the chrysanthemum flavonoids group, there was no statistical difference in the Apo-A1 content in the serum of the luteolin group and the simvastatin group (P>0.05).

[0111] The content of Apo-B in the serum of the hyperlipidemia model group was higher than that of the normal control group, although there was an increasing trend but no statistical difference (P>0.05). After intervention with chrysanthemum flavonoids, luteolin and simvastatin, the content of Apo-B in the serum of the chrysanthemum flavonoids group was significantly decreased (P<0.05), and there was no statistical difference in the serum Apo-B between the luteolin and simvastatin groups (P>0.05), and there was no statistical difference between the normal group (P>0.05). Compared with the chrysanthemum flavonoids group, the content of Apo-B in the luteolin group and the simvastatin group was significantly increased (P<0.05). There was no statistical difference in Apo-B / Apo-A1 among the groups (P>0.05).

[0112] Luteolin can significantly improve the body weight, organ coefficient, and body fat rate of SD rats induced by a high-fat diet; significantly reduce the levels of TC, TG, and LDL-C in the serum of hyperlipidemic SD rats, thereby alleviating the increase in blood lipid levels; Luteolin can significantly improve the degree of liver fat degeneration and restore the normal morphology and number of peritesticular fat adipocytes; these results reveal that Luteolin has a good lipid-lowering effect. The effect of Luteolin on improving liver fatty degeneration may be achieved by inhibiting lipid peroxidation and increasing the activity of antioxidant enzymes.

[0113] Example 2

[0114] A total of 44 subjects were recruited in this study, and 37 subjects were identified after population screening according to the inclusion and exclusion criteria. Then the subjects were randomly divided into a control group (n=18) and a chrysanthemum group (n=19) according to basic conditions (fasting blood glucose lipids, gender, etc.). This study was approved by the Clinical Research Ethics Committee of Zhongda Hospital Affiliated to Southeast University (2020ZDSYLL263-P01) and registered with the China Clinical Trial Registry (ChiCTR2100044946). All subjects gave informed consent. The inclusion and exclusion criteria are as follows:

[0115] Inclusion criteria: 1) healthy people with BMI between 18.5-23.9, and fasting TG <1.70mmol / L, TC <5.72mmol / L, HDL-C>0.91mmol / L, LDL-C <3.6mmol / L; 2) people without other serious metabolic or chronic diseases. Exclusion criteria: 1) people with chronic diseases such as diabetes and hypertension; 2) people with gastrointestinal diseases; 3) people with severe smoking and drinking.

[0116] The baseline information of the two groups is shown in Table 6. There were 18 people in the control group and 19 people in the Huangju group. There was no statistically significant difference in gender, age, height and other data between the two groups.

[0117] Table 6 Baseline information of the two groups (x±s)

[0118]

[0119] Note: The results are mean ± standard deviation

[0120] For the convenience of eating, this study adopted the liquid high-fat load diet used in previous studies by the research team (fat accounts for 50% of total energy, carbohydrates 35%, protein 15%). The dietary composition (based on 60kg / person) is shown in Table 3:

[0121] Table 7: Dietary Composition

[0122]

[0123] High-fat load meal production process: mixed raw materials + purified water - heating - shearing (80°C, 2000r / min) - homogenization - pasteurization.

[0124] Chrysanthemum flavonoids extract was processed in advance according to "Part I". The amount of high-fat load meal was based on the subject's body weight and was given at 18 kcal / kg·bw (body weight). The subjects arrived at the hospital early the next morning after fasting for 12 hours. After collecting fasting blood, the meal was distributed according to the subject's weight, and the subject was required to drink it within 5 minutes and start timing. Blood samples were collected 0.5, 1, 2, 4, 6, and 8 hours after the meal and immediately centrifuged to obtain serum for testing. During the blood sample collection period, the subjects were not allowed to exercise vigorously and could drink water in moderation.

[0125] Measurement indicators: blood sugar, four blood lipids, superoxide dismutase (SOD), proglutathione (GSH) and total antioxidant capacity (T-AOC).

[0126] Test results:

[0127] Table 8 Changes in serum GLU levels after meals

[0128]

[0129] Note: *Compared with the control group, #Compared with fasting (*, #p<0.05, **, ##p<0.01)

[0130] The changes in serum GLU levels of the subjects after the meal are shown in Table 8. After the SW test, the data of each group obeyed the normal distribution. The result of the sphericity test, MachlyW=0.013, was significant (p<0.001), and did not meet the sphericity test. Therefore, the correction results in the univariate analysis of variance "Greenhouse-Geisser" shall prevail. The results of repeated measures analysis of variance are shown in Table 6. The results of the simple effect test of the group showed that the serum GLU level of the Huangju group was significantly lower than that of the control group 1h after the meal (p<0.01), and the serum GLU level of the Huangju group was significantly higher than that of the control group 4, 6, and 8h after the meal (p<0.05). The results of the simple effect of measurement time showed that the simple effect of measurement time in the control group and the Huangju group was extremely significant (F=12.72, p<0.001, η2=0.72; F=5.22, p=0.001, η2=0.51). Multiple comparisons found that the serum GLU level in the control group was significantly higher than that in the fasting state within 2 hours after the meal (p<0.01); the serum GLU level in the royal chrysanthemum group increased significantly 0.5 hours after the meal (p<0.01), and then began to decrease. Table 8 shows the trend of postprandial serum GLU levels in each group. The maximum change value of postprandial serum GLU levels and postprandial PR in the control group and royal chrysanthemum group were statistically significant.

[0131] Chrysanthemum extract affects serum GLU levels in a short period of time and prevents the increase of serum blood lipid levels at different time points, thereby enhancing the body's antioxidant capacity.

[0132] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0133] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A chrysanthemum extract, characterized in that The chrysanthemum extract contains chrysanthemum flavonoids, and the main components of the chrysanthemum flavonoids include: Luteolin-7-O-rutinoside, whose molecular formula is C 27 H 30 O 15 , the structural formula is: Luteolin-7-O-glucoside, whose molecular formula is C 21 H 20 O 11 , the structural formula is: Luteolin-7-O-glucuronide, whose molecular formula is C 21 H 18 O 12 , the structural formula is: Luteolin-7-O-6"-malonyl glucoside, whose molecular formula is C 24 H 22 O 14 , the structural formula is: Luteolin-7-O-2"-acetyl glucuronide, whose molecular formula is C 23 H 20 O 13 , the structural formula is: Luteolin-7-O-6"-acetyl glucuronide, whose molecular formula is C 23 H 22 O 12 , the structural formula is:

2. A method for preparing a chrysanthemum extract, characterized in that: The preparation method comprises the following steps: S1, removing impurities from the picked chrysanthemums to retain the complete inflorescence, and then drying them to obtain dried chrysanthemums; S2, crushing and grinding the dried chrysanthemum, and then sieving to obtain chrysanthemum powder; S3, using an ethanol solution to fully soak the chrysanthemum powder, and then subjecting the chrysanthemum powder to ultrasonic oscillation to obtain a chrysanthemum powder soaking solution; S4, extracting the chrysanthemum powder soaking solution in a water bath to obtain an extract; S5, filtering the extract to remove impurities, and then concentrating the extract to 1% of the volume of the original solution to obtain a concentrated solution; S6. freeze-drying the concentrated solution to obtain a crude chrysanthemum extract; S7. Dissolve the crude chrysanthemum extract in water to prepare a chrysanthemum solution, adjust the pH of the chrysanthemum solution to 2.5, and then flow into a macroporous adsorption resin at a flow rate of 1.00 mL / min; use an ethanol solution to analyze the macroporous adsorption resin adsorbed with the chrysanthemum solution; and obtain a chrysanthemum extract after concentrating and freeze-drying the effluent.

3. The method for preparing a chrysanthemum extract according to claim 2, characterized in that: In step S1, the drying method adopts a step-by-step low-temperature drying method, and the temperature is set to 25°C - 35°C - 45°C - 35°C - 25°C.

4. The method for preparing a chrysanthemum extract according to claim 2, characterized in that: In step S2, a sieve with a mesh size of 200 to 250 is used for screening.

5. The method for preparing a chrysanthemum extract according to claim 2, characterized in that: In step S4, the material-liquid ratio of the chrysanthemum powder and the clean water is 1:40; the water bath temperature is 85° C. to 100° C., and the water bath time is 45 to 60 minutes.

6. The method for preparing a chrysanthemum extract according to claim 2, characterized in that: The concentration of the ethanol solution used in steps S3 and S7 is 70%.

7. The method for preparing a chrysanthemum extract according to claim 2, characterized in that: The concentration of the chrysanthemum solution in step S7 is 3.60 mg / mL, and the volume of the chrysanthemum solution is twice the volume of the macroporous adsorption resin; when using ethanol solution for analysis, the volume of the ethanol solution is twice the volume of the macroporous adsorption resin.

8. Use of the chrysanthemum extract as claimed in claim 1 as a raw material in the preparation of lipid-lowering drugs.

9. Use of the chrysanthemum extract as claimed in claim 1 as a raw material in the preparation of a drug for enhancing the body's antioxidant capacity.

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