CN115956630A

CN115956630A - Compound magnolia bark extract, preparation method and quality control method thereof

Info

Publication number: CN115956630A
Application number: CN202211666017.1A
Authority: CN
Inventors: 吕剑波; 马亚; 刘起军; 王亚利
Original assignee: Sichuan Hengtong Animal Protection Biotechnology Co ltd
Current assignee: Sichuan Hengtong Animal Protection Biotechnology Co ltd
Priority date: 2022-12-23
Filing date: 2022-12-23
Publication date: 2023-04-14

Abstract

The invention discloses a compound magnolia bark extract, which comprises the following raw materials in parts by weight: 1-60 parts of mangnolia officinalis, 3-50 parts of cherokee rose fruit meat, 1-60 parts of scorched hawthorn fruit, 4-63 parts of scorched medicated leaven, 1-60 parts of scorched malt, 82-120 parts of ginger and 90-101 parts of Chinese date meat. The invention also discloses a preparation method of the compound magnolia bark extract, and a quality control method of the compound magnolia bark extract, which comprises a detection method of cherokee rose fruit flesh and a detection method of a characteristic map of magnolia bark. The invention adopts a semi-bionic extraction method, simulates the physiological conditions of the gastrointestinal tract of the weaned piglets, ensures that the obtained effective components are more beneficial to the weaned piglets, and avoids the adverse effects caused by the obvious difference between the weaned piglets and the physiological conditions (pH and enzyme types) of the gastrointestinal tract of the weaned piglets and the adult piglets.

Description

Compound magnolia bark extract, preparation method and quality control method thereof

Technical Field

The invention relates to the field of feed additives, in particular to a compound magnolia officinalis extract, a preparation method and a quality control method thereof.

Background

Probiotics refer to microorganisms beneficial to the health of organisms, and as of 2019, 35 or subspecies of probiotics which can be used in the field of food are released in China. The modern Chinese medicine fermentation technology inherits the processing method of the traditional Chinese medicine and is combined with the modern bioengineering technology. The probiotic fermented traditional Chinese medicine has the advantages of reducing toxic and side effects, generating new drug effect substances, enlarging indications, saving traditional Chinese medicine material resources, reducing cost and the like. The solid fermentation does not need to sterilize a simple nutrient medium, belongs to an open system, has easy operation of fermentation conditions such as temperature, humidity, pH value, ventilation and the like, but still has ineffective components which are difficult to absorb and digest after the fermentation, and is not beneficial to the modernization of traditional Chinese medicines.

The semi-bionic extraction method is proposed in 1995 by Zhangmegawang and Sunxuimei, which are universities of traditional Chinese medicines in Shandong, and is currently applied to various traditional Chinese medicine preparations, such as Shaoyan pain-relieving granules, hantongding effervescent granules, coptis chinensis detoxifying granules and the like. The method simulates the transportation process of oral drugs in the gastrointestinal tract, and adopts acidic water and alkaline water with selected pH value to extract sequentially and continuously. The traditional semi-bionic extraction method adopts water decoction or heating reflux extraction, has high extraction temperature and large difference with the gastrointestinal tract environment, mostly does not consider the physiological conditions of pH, enzyme types, gastrointestinal tract peristalsis capability and the like of the digestive system of young animals, and has complex extracted components.

The Chinese veterinary pharmacopoeia adopts the accompanying control of national standard substances (fructus Rosae Laevigatae reference medicinal materials) and thin-layer chromatography to identify fructus Rosae Laevigatae medicinal materials. The concentrated sulfuric acid is strong acid, the trichloromethane is easy to produce poison, the concentrated sulfuric acid and the trichloromethane are not friendly to operators and the environment, the thin-layer plate needs to be heated at 105 ℃ after the color developing agent is sprayed, and the operation is complicated.

The quality control of the magnolia officinalis in the existing compound preparation mainly comprises thin-layer chromatography identification and content determination of magnolol and honokiol. The magnolia officinalis contains a plurality of chemical components, and the quality of a single component is difficult to reflect.

The sow is long in birth process, so that a lot of adverse effects are caused, especially dead fetus and weak piglets are easy to generate, the postnatal physique of the piglets is poor, the growth is slow, and the survival rate is low. The sow is also greatly damaged due to the over-long labor process, for example, the sow with the dystocia has the diseases of parturition syndrome, postpartum hyperpyrexia, uterine bleeding, birth canal bleeding, poor mental state, slow recovery, metritis infection and the like, and even the sow can be directly eliminated. Therefore, feasible measures for shortening the labor of the sows are urgently needed to reduce the loss of the producers caused by the overlong labor of the sows, and the method has great significance for the pig raising industry.

Disclosure of Invention

The invention aims to provide a compound magnolia bark extract, a preparation method and a quality control method thereof, which are used for solving the technical problem of overlong labor course of sows in the prior art.

In order to achieve the purpose, one embodiment of the invention provides a compound magnolia bark extract, which comprises the following raw materials in parts by weight:

1 to 60 portions of magnolia officinalis, 3 to 50 portions of cherokee rose fruit flesh, 1 to 60 portions of scorched hawthorn fruit,

4 to 63 portions of charred medicated leaven, 1 to 60 portions of charred malt, 82 to 120 portions of ginger,

90-101 parts of jujube meat.

In one preferred scheme of the invention, the compound magnolia bark extract comprises the following raw materials in parts by weight: 52-57 parts of mangnolia officinalis, 29-33 parts of cherokee rose fruit meat, 12-18 parts of scorched hawthorn fruit, 35-36 parts of scorched medicated leaven, 26-28 parts of scorched malt, 102-103 parts of ginger and 96 parts of Chinese date meat.

Based on the compound magnolia bark extract disclosed by the invention, the invention also discloses a preparation method of the compound magnolia bark extract, which comprises the following steps:

step (1): pulverizing charred Massa Medicata Fermentata into fine powder, pulverizing cortex Magnolia officinalis, fructus Rosae Laevigatae, charred fructus crataegi and charred fructus Hordei Germinatus, and making into raw material micropowder;

step (2): adding water into the ginger and jujube pulp, and juicing to obtain ginger and jujube juice;

and (3): uniformly mixing the fine powder of the charred medicated leaven prepared in the step (1), the micro powder of the raw materials and the ginger and jujube juice prepared in the step (2) to obtain a fermentation substrate;

and (4): adding composite probiotic powder, uniformly mixing with a fermentation substrate, and fermenting to obtain a fermentation product;

and (5): adding additives into the fermentation product prepared in the step (4) for multiple times, stirring and extracting, filtering after extraction is finished, centrifuging filtrate, and concentrating to obtain a fermentation raw material concentrated solution;

and (6): and (3) taking the concentrated fermentation raw material solution prepared in the step (5), adjusting the pH, passing through a macroporous adsorption resin column, eluting with water and ethanol in sequence, collecting the eluent, concentrating the eluent, drying, crushing, mixing totally, and subpackaging to obtain the compound magnolia bark extract.

In one preferable scheme of the invention, the composite probiotic powder in the step (4) comprises 1 part by mass of monascus anka, 0.8-3.6 parts by mass of lactobacillus plantarum and 1-1.7 parts by mass of bifidobacterium bifidum, and the mass ratio of the inoculation amount of the composite probiotic powder to the fermentation substrate is 1:46-130.

In one preferable scheme of the invention, the fermentation raw material in the step (5) needs to be stirred and extracted for three times, wherein the additive added in the first stirring and extraction is a mixture of a medium and enzyme, the medium is lactic acid, hydrochloric acid, citric acid and water, the enzyme is pepsin, rennin and lipase, and the extraction time is 0.5h; the additive added in the second stirring extraction is a mixture of a medium and enzyme, wherein the medium is sodium bicarbonate and water, the enzyme is lactase, amylase and trypsin, and the extraction time is 1h; the additive added in the third stirring extraction is a medium, wherein the medium is sodium carbonate and water, and the extraction time is 2h.

The invention also discloses a quality control method of the compound magnolia bark extract, which comprises a detection method of cherokee rose fruit flesh and a detection method of a magnolia bark characteristic map.

In a preferred embodiment of the present invention, the method for detecting cherokee rose-hip comprises the following steps:

a, step a: preparing a test solution: grinding 1g of the product, extracting with solvent, filtering, evaporating filtrate, dissolving residue with solvent, and diluting to obtain sample solution;

step b: preparation of control solutions: extracting fructus Rosae Laevigatae 1g with solvent, filtering, evaporating filtrate, dissolving residue with solvent, and diluting to obtain reference solution;

step c: and (3) determination: sucking the test solution and the reference solution, respectively dropping on the same silica gel G thin layer plate, developing in developing agent, taking out, air drying, and inspecting.

In one preferable scheme of the invention, the extraction solvent in the step a is ethyl acetate, and the extraction mode is ultrasonic extraction; in the step b, the extraction solvent is ethyl acetate, and the extraction mode is any one of ultrasonic extraction and heating reflux extraction; in the step c, the developing solvent is a mixture of petroleum ether, ethyl acetate and formic acid, wherein the volume ratio of the petroleum ether to the ethyl acetate to the formic acid is 100.

In one preferable scheme of the invention, the detection method of the magnolia officinalis characteristic spectrum comprises the following steps:

step a: preparation of reference solutions: weighing honokiol reference substance, and adding solvent to obtain reference solution;

step b: preparing a test solution: weighing compound cortex Magnolia officinalis extract, placing in conical flask with plug, adding acetone, ultrasonic treating, filtering, evaporating filtrate with water bath, dissolving residue in water, passing through D101 macroporous adsorbent resin column, sequentially eluting with water and ethanol, collecting ethanol eluate, filtering, and collecting filtrate to obtain sample solution;

step c: and (3) determination: and respectively sucking the reference substance solution and the reference substance solution, and injecting the reference substance solution and the reference substance solution into a liquid chromatograph for measurement.

In a preferred embodiment of the present invention, the chromatographic conditions in step c are: the chromatographic column is C18, the filler is octadecylsilane chemically bonded silica, the mobile phase is a mixed solution of methanol and 0.4% formic acid, wherein the methanol is a mobile phase A, the 0.4% formic acid solution is a mobile phase B, the flow rate is 1ml/min, the column temperature is 30 ℃, and the detection wavelength is 230nm.

In conclusion, the beneficial effects of the invention are as follows:

1. the invention ensures that the effective components are dissolved out more completely and the conditions are milder through the solid fermentation of the probiotics, thereby avoiding the damage of heat-sensitive effective substances.

2. The invention adopts a semi-bionic extraction method, simulates the physiological conditions of the gastrointestinal tract of the weaned piglets, ensures that the obtained effective components are more beneficial to the weaned piglets, and avoids the adverse effects caused by the obvious difference between the weaned piglets and the physiological conditions (pH and enzyme types) of the gastrointestinal tract of the weaned piglets and the adult piglets.

3. The invention adopts low-speed stirring extraction, simulates the characteristic of weak gastrointestinal peristalsis function of the weaned piglets, and prevents impurities from dissolving out.

4. The extracting solution is separated and purified by macroporous absorption resin, so that the effective components (such as total sugar and total flavone) are enriched, and the effectiveness of the preparation is ensured.

5. The invention can shorten the labor process of sows, has obvious curative effect on treating diarrhea of weaned piglets in the inland river, and has the pharmacological actions of enhancing the immunity and adjusting the gastrointestinal tract movement.

6. The invention adopts thin-layer chromatography to identify the cherokee rose fruit flesh in the compound magnolia bark extract, and the method has strong specificity, simple and convenient operation and good reproducibility.

7. The invention adopts high performance liquid chromatography to establish the characteristic spectrum of the magnolia officinalis in the compound magnolia officinalis extract, and can be used for comprehensively evaluating the quality of the magnolia officinalis.

Drawings

FIG. 1 is a schematic diagram of the TLC identification of a portion of Cherokee rose flesh from Magnolia bark extract (9 pilot runs in total) in accordance with one embodiment of the present invention;

FIG. 2 is a graph of thin layer chromatography identification of another portion of FIG. 1 in accordance with an embodiment of the present invention;

FIG. 3 is a diagram of the test result of the thin layer chromatography identification method of the Magnolia bark extract according to the item of "Cherokee Rose fruit" in the second part of 2020 animal pharmacopoeia of the Compound Houpo of the present invention;

FIG. 4 is a diagram of the inspection result of the specificity of Magnolia bark by the method for detecting the characteristic spectrum of Magnolia bark extract;

FIG. 5 is an HPLC control profile of Magnolia officinalis in the established Magnolia officinalis extract according to one embodiment of the present invention;

FIG. 6 is a histogram overlay of Magnolia officinalis characteristics in 15 batches of Magnolia bark extract, in accordance with one embodiment of the present invention.

Detailed Description

The compound magnolia bark extract comprises the following raw materials in parts by weight: 1-60 parts of mangnolia officinalis, 3-50 parts of cherokee rose fruit meat, 1-60 parts of scorched hawthorn fruit, 4-63 parts of scorched medicated leaven, 1-60 parts of scorched malt, 82-120 parts of ginger and 90-101 parts of fresh jujube meat.

Preferably, the compound magnolia bark extract comprises the following raw materials in parts by weight: 52-57 parts of mangnolia officinalis, 29-33 parts of cherokee rose fruit meat, 12-18 parts of scorched hawthorn fruit, 35-36 parts of scorched medicated leaven, 26-28 parts of scorched malt, 102-103 parts of ginger and 96 parts of fresh Chinese date meat;

a preparation method of a compound magnolia bark extract comprises the following steps:

step (1): pulverizing the charred Massa Medicata Fermentata at low temperature into fine powder, and micronizing cortex Magnolia officinalis, fructus Rosae Laevigatae, charred fructus crataegi and charred fructus Hordei Germinatus to obtain superfine powder;

step (2): adding water into rhizoma Zingiberis recens and fructus Jujubae meat, and squeezing to obtain rhizoma Zingiberis recens and fructus Jujubae juice;

and (3): taking the fine powder of the charred medicated leaven prepared in the step (1), the superfine powder of the raw materials and the ginger and jujube juice prepared in the step (2), adding probiotics, mixing uniformly to obtain a fermentation substrate, wherein the amount of the added ginger and jujube juice is 46%,

and (4): adding a proper amount of composite probiotic powder, uniformly mixing with a fermentation substrate, and fermenting to obtain a fermentation product, wherein the composite probiotic powder comprises 1 part by mass of Monascus anka AS 3.782 with a CCTCC number of AF 93208, 0.8-3.6 parts by mass of Lactobacillus plantarum with a CCTCC number of AB 2010210, 1-1.7 parts by mass of Bifidobacterium bifidum (Bifidobacterium bifidum ATCC29521 with a CCTCC number of AB 94055), and the mass ratio of the inoculation amount of the composite probiotic powder to the fermentation substrate is 1:46 to 130 ℃, the fermentation temperature is 21 to 37 ℃, the fermentation temperature is preferably 33 to 34 ℃, and the fermentation time is 72 to 120 hours;

and (5): adding lactic acid, hydrochloric acid, citric acid and water into the fermentation product obtained in the step (4) (and adjusting the pH value to 4.6), adding pepsin, chymosin and lipase, extracting for 0.5h at 37 ℃ under stirring (the rotation speed is 45 rpm), filtering, adding sodium bicarbonate and water into the dregs (and adjusting the pH value to 7.2), adding lactase, amylase and trypsin into the dregs, extracting for 1h at 37 ℃ under stirring (the rotation speed is 45 rpm), filtering, adding sodium carbonate and water into the dregs (and adjusting the pH value to 6.1), extracting for 2h at 37 ℃ under stirring (the rotation speed is 45 rpm), and filtering; centrifuging and concentrating the filtrate to obtain a concentrated solution of the fermentation raw material;

and (6): and (3) taking the concentrated fermentation raw material solution prepared in the step (5), adjusting the pH value to be neutral, passing through an AB-8 type macroporous adsorption resin column, eluting with water and 80% ethanol in sequence, collecting the eluent, concentrating the eluent, drying, crushing, mixing totally, and subpackaging to obtain the compound magnolia bark extract.

A quality control method of compound cortex Magnolia officinalis extract comprises fructus Rosae Laevigatae detection method and cortex Magnolia officinalis characteristic chromatogram detection method.

The detection method of the cherokee rose flesh comprises the following steps:

a, step a: preparing a test solution: grinding 1g of the product, adding 50ml of ethyl acetate solvent, performing ultrasonic extraction for 20min, filtering, evaporating filtrate, dissolving residue with ethyl acetate, and diluting to 0.5ml to obtain sample solution;

step b: preparation of control solutions: collecting fructus Rosae Laevigatae control medicinal material 1g, adding 50ml ethyl acetate solvent, performing ultrasonic extraction or heating reflux, preferably heating reflux extraction for 1h, filtering, evaporating filtrate, dissolving residue with ethyl acetate, and diluting to 0.5ml to obtain control solution;

step c: and (3) determination: sucking sample solution and reference solution 5ul, respectively dropping on the same silica gel G thin layer plate, developing in developing agent to 8cm, taking out, air drying, inspecting under ultraviolet lamp (365 nm), and displaying a same yellow fluorescent spot in the sample chromatogram at the position corresponding to the control chromatogram; the developing agent is a mixture of petroleum ether, ethyl acetate and formic acid, and the mass ratio of the petroleum ether to the ethyl acetate to the formic acid is 100.

The detection method of the magnolia officinalis feature map comprises the following steps:

step a: preparation of reference solutions: precisely weighing honokiol reference substance, and adding methanol to obtain reference substance solution containing 30.26ug of honokiol per 1 ml;

step b: preparing a test solution: precisely weighing 0.8g of compound cortex magnoliae officinalis extract, placing in a conical flask with a plug, adding 25ml of acetone, performing ultrasonic treatment for 20min, filtering, evaporating the filtrate in a water bath, dissolving the residue in 50ml of water, passing through a D101 macroporous adsorption resin column (the inner diameter is 1cm, the column height is 15 cm), sequentially eluting with 50ml of water and 50ml of 65% ethanol, collecting 65% ethanol eluate, filtering with a microporous membrane, and collecting the filtrate to obtain a sample solution;

step c: and (3) determination: respectively sucking reference substance solution and reference substance solution, and injecting into a liquid chromatograph for determination;

the high performance liquid chromatography is adopted for analysis, and the chromatographic conditions are as follows:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filler, and particularly, a chromatographic column is Wondasil C18super (the column is 250mm long, the inner diameter is 4.6mm, and the particle size is 5 um);

mobile phase: methanol is taken as a mobile phase A, and 0.4% formic acid solution is taken as a mobile phase B;

gradient elution: gradient elution was performed as specified in the following table, where% in the table refers to volume percent:

time (minutes)	Mobile phase A (%, v/v)	Mobile phase B (%, v/v)	Elution is carried out
				0→17	11	89	Equal degree
17→23	11→17	89→83	Linear gradient
				23→41	17→23	83→77	Linear gradient
41→64	23→58	77→42	Linear gradient
				64→77	58→64	42→36	Linear gradient
77→79	64	36	Equal degree
				79→85	64→94	36→6	Linear gradient
85→95	6	6	Equal degree

Flow rate: 1ml/min

Column temperature: 30 deg.C

Detection wavelength: 230nm

The reference characteristic spectrum of the compound magnolia bark extract comprises 16 characteristic peaks which are respectively 1 to 16. Wherein, the peak 9 corresponding to the honokiol reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time of each peak is as follows:

peak number	Relative retention time	RSD(％)
			1	0.14	1.84
2	0.24	1.30
			3	0.36	1.15
4	0.44	1.59
			5	0.52	0.91
6	0.59	0.81
			7	0.86	0.57
8	0.97	1.04
			9	1.00	0.00
10	1.02	0.54
			11	1.03	0.59
12	1.05	0.61
			13	1.07	0.49
14	1.08	0.38
			15	1.21	0.54
16	1.33	0.51

Example 1

The compound magnolia bark extract comprises the following raw materials in parts by weight:

52 parts of mangnolia officinalis, 33 parts of cherokee rose fruit meat, 18 parts of scorched hawthorn fruit, 35 parts of scorched medicated leaven, 26 parts of scorched malt, 102 parts of ginger and 96 parts of fresh jujube meat.

step (1): taking 35g of charred medicated leaven, pulverizing into fine powder at low temperature, taking 4 kinds of medicines such as 52g of mangnolia officinalis, 33g of cherokee rose pulp, 18g of charred hawthorn, 26g of charred malt and the like, carrying out superfine grinding to prepare raw material superfine powder, and controlling the grain diameter D (v, 0.9) of the superfine powder to be less than or equal to 5um;

step (2): and adding a proper amount of water into 102g of ginger and 96g of jujube meat, and squeezing the fresh ginger and jujube meat to obtain ginger and jujube juice.

and (4): adding a proper amount of composite probiotic powder, uniformly mixing with a fermentation substrate, and fermenting to obtain a fermentation product; the composite probiotic powder comprises 1 part of Monacus anka AS 3.782 with the CCTCC number of AF 93208, 1 part of Lactobacillus plantarum (with the CCTCC number of AB 2010210), 0.8 part of Bifidobacterium bifidum (with the CCTCC number of AB 201021521), and 1 part of Bifidobacterium bifidum (with the CCTCC number of AB 94055) by mass, wherein the mass ratio of the inoculation amount of the composite probiotic powder to the fermentation substrate is 1:110, the fermentation temperature is 33 ℃, and the fermentation time is 72 hours;

and (5): adding lactic acid, hydrochloric acid, citric acid and water into the fermentation product obtained in the step (4) (and adjusting the pH value to 4.6), adding pepsin, chymosin and lipase, extracting for 0.5h at 37 ℃ under stirring (the rotation speed is 45 rpm), filtering, adding sodium bicarbonate and water into the dregs (and adjusting the pH value to 7.2), adding lactase, amylase and trypsin into the dregs, extracting for 1h at 37 ℃ under stirring (the rotation speed is 45 rpm), filtering, adding sodium carbonate and water into the dregs (and adjusting the pH value to 6.1), extracting for 2h at 37 ℃ under stirring (the rotation speed is 45 rpm), and filtering. Centrifuging the filtrate, and concentrating to obtain concentrated solution of fermentation raw material;

and (6): and (3) taking the concentrated solution of the fermentation raw material in the step (5), adjusting the pH value to be neutral, passing through an AB-8 type macroporous adsorption resin column, eluting with water and 80% ethanol in sequence, collecting eluent, concentrating, drying, crushing, mixing, and subpackaging to obtain the finished product.

Example 2

The thin-layer chromatography identification experiment research of the cherokee rose fruit meat in the compound magnolia officinalis extract comprises the following steps:

1: standard substance and sample

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2: screening of test conditions

2.1 examination of the amount of formic acid

As a result: indicating that only 0.1 formic acid was added.

2.2 screening of developer species

2.3 selection of test article preparation method

Preparing a magnolia bark extract test sample solution (methanol ultrasonic extraction and ethyl acetate extraction): weighing cortex Magnolia officinalis extract 1g, adding methanol 50ml, ultrasonic treating for 20min, filtering, evaporating filtrate to dryness, dissolving residue with water 50ml, extracting with ethyl acetate for 3 times, each time 30ml, mixing ethyl acetate solutions, evaporating in water bath, dissolving residue with ethanol, and diluting to 1 ml.

Preparing a magnolia officinalis extract test sample solution (ethyl acetate ultrasonic extraction): weighing cortex Magnolia officinalis extract 1g, grinding, adding ethyl acetate 50ml, ultrasonic treating for 20min, filtering, evaporating filtrate, dissolving residue in ethyl acetate, and diluting to 0.5ml to obtain test solution.

Preparation of cherokee rose fruit reference medicinal material solution (methanol ultrasonic extraction, ethyl acetate extraction): weighing fructus Rosae Laevigatae reference medicinal material 1g, adding methanol 50ml, ultrasonic processing for 20min, filtering, evaporating filtrate to dryness, dissolving residue with water 50ml, extracting with ethyl acetate for 3 times, each time 30ml, mixing ethyl acetate solutions, evaporating in water bath, dissolving residue with ethanol, and diluting to 1 ml.

Preparation of cherokee rose fruit reference medicinal material solution (ethyl acetate ultrasonic extraction): weighing fructus Rosae Laevigatae reference material 1g, adding ethyl acetate 50ml, ultrasonic treating for 20min, filtering, evaporating filtrate, adding ethyl acetate into residue to dissolve and dilute to 0.5ml, and using as sample solution.

Preparation of cherokee rose fruit reference medicinal material solution (heating reflux extraction by ethyl acetate): weighing fructus Rosae Laevigatae reference material 1g, adding ethyl acetate 50ml, heating and refluxing for extraction for 1h, filtering, evaporating filtrate, adding ethyl acetate into residue to dissolve and dilute to 0.5ml, and using as reference material solution.

The results are shown in table 1:

table 1: examination result of test sample preparation method

As can be seen from Table 1, the fructus Rosae Laevigatae reference drug is extracted by heating and refluxing with ethyl acetate, and the thin-layer chromatography of the test sample prepared by the compound cortex Magnolia officinalis extract by ethyl acetate ultrasonic extraction is good.

3: test methods and results

Preparation of a test solution: grinding 1g of the product, adding 50ml of ethyl acetate, performing ultrasonic treatment for 20min, filtering, evaporating the filtrate, dissolving the residue with ethyl acetate, and diluting to 0.5ml to obtain a sample solution.

Preparation of reference drug solution: and adding 50ml of ethyl acetate into 1g of fructus Rosae Laevigatae control medicinal material, extracting under reflux for 1h, filtering, evaporating the filtrate, dissolving the residue with ethyl acetate, and diluting to 0.5ml to obtain control solution.

Preparation of negative solution lacking cherokee rose flesh: collecting negative 1g of fructus Rosae Laevigatae pulp, grinding, adding ethyl acetate 50ml, ultrasonic treating for 20min, filtering, evaporating filtrate, dissolving residue with ethyl acetate, and diluting to 0.5ml to obtain negative solution.

Thin layer chromatography assay: according to a thin-layer chromatography test (appendix 0502 of Chinese veterinary pharmacopoeia), 5ul of each of the two solutions is absorbed, the two solutions are respectively spotted on the same silica gel G thin-layer plate, petroleum ether (60-90 ℃), ethyl acetate-formic acid (10.1) are used as a developing agent, the mixture is spread to about 8cm, and the mixture is taken out, dried and placed under an ultraviolet lamp (365 nm) for inspection.

As a result: in the test sample chromatogram of the 9 batches of compound magnolia bark extract pilot-plant sample, the same yellow fluorescent spot is displayed at the position corresponding to the Cherokee rose fruit reference medicinal material chromatogram, and the negative is non-interference, which indicates that the method has strong specificity and good repeatability, and the result is shown in figure 1 and figure 2, wherein 1-3 are compound magnolia bark extracts (the pilot-plant numbers are 20220301, 20220302 and 20220303 in sequence); 4 and 9 are fructus Rosae Laevigatae reference materials; 5-6 are compound cortex magnoliae officinalis extracts (the pilot plant numbers are 20220304 and 20220305 in sequence); 7-8 are magnolia officinalis extracts (the pilot plant numbers are 20220401 and 20220402 in sequence); 10-11 are compound magnolia bark extracts (the pilot plant numbers are 20220501 and 20220502 in sequence); 12 is negative for cherokee rose-lacking flesh.

Example 3

The method for establishing the Magnolia officinalis characteristic spectrum in the compound Magnolia officinalis extract comprises the following specific tests:

1: material

Magnolia officinalis (magnolia officinalis) reference drug (lot No. 121285-201303, china institute for food and drug testing); magnolol reference substance (China institute for food and drug testing, purity 99.0%, batch No. 110729-202015); honokiol reference substance (China institute for food and drug testing, purity 99.8%, batch No. 110730-201915);

collecting 15 batches of Magnolia bark decoction pieces, the dried bark, root bark and branch bark are identified as Magnolia officinalis of Magnoliaceae, magnolia officinalis of Magnolia officinalis Rehd. The corresponding finished products are prepared according to the prescription and the process of the invention, and the detailed information is shown in table 2.

Table 2: information of source and finished product of Magnolia officinalis decoction pieces (15 batches)

Note: wherein 7 batches (batches 20220101, 20220102, 20220103, 20210104, 20220220220201, 20220220202 and 20220220220220203) are pilot plant samples of the laboratory, and 8 batches (batches 220301, 20220302, 20220303, 20220304, 20220401, 20220402, 20220501 and 20220502) are pilot plant samples of the company.

2: chromatographic conditions

LC-20A Shimadzu high performance liquid chromatograph (SPD-20A type ultraviolet visible light detector, SIL-20AxR type autosampler, column oven CTO-20AC, DGU-20A5R degasser); wondasil C18super (4.6X250mm, 5um, S/N:9F 3706-04); column temperature: 30 ℃; flow rate: 1ml/min; the sample amount is 10ul; gradient elution was performed as specified in table 3 with methanol as mobile phase a and 0.4% formic acid solution as mobile phase B; the detection wavelength was 230nm.

TABLE 3 gradient elution procedure

3: preparation of sample solution

Preparing a magnolia officinalis decoction piece test solution: precisely weighing cortex Magnolia officinalis decoction pieces about 0.1g, placing into conical flask with plug, precisely adding 60% methanol 50ml, ultrasonic treating for 20min, cooling, weighing, supplementing with 60% methanol, shaking, filtering with microporous membrane, and collecting filtrate

Preparing a reference substance solution of a magnolia officinalis reference medicinal material: precisely weighing about 0.1g of cortex Magnolia officinalis (Magnolia officinalis) as control medicinal material powder, placing into conical flask with plug, precisely adding 50ml of 60% methanol, ultrasonically treating for 20min, cooling, weighing, supplementing lost weight with 60% methanol, shaking, filtering with microporous membrane, and collecting filtrate.

Preparation of honokiol reference solution: accurately weighing 15.16mg of honokiol reference substance (China institute for food and drug inspection, purity 99.8%, batch No. 110730-201915), placing in a 100ml brown measuring flask, adding methanol to dissolve and dilute to scale, accurately weighing 2ml, placing in a 10ml brown measuring flask, adding methanol to dissolve and dilute to scale, and making into the product with concentration of 30.26ug/ml.

Preparation of magnolol reference solution: accurately weighing magnolol control substance 20.75mg, placing in 50ml brown measuring flask, adding methanol to dissolve and dilute to scale, accurately weighing 1ml, placing in 10ml brown measuring flask, adding methanol to dissolve and dilute to scale to obtain the final product with concentration of 41.09ug/ml.

Preparation of a compound magnolia bark extract test solution: taking about 0.8g of compound magnolia bark extract, precisely weighing, placing in a conical flask with a plug, adding 25ml of acetone, carrying out ultrasonic treatment for 20min, filtering, evaporating the filtrate in a water bath, adding 50ml of water into the residue, carrying out ultrasonic treatment to dissolve the residue, passing through a D101 macroporous adsorption resin column (the inner diameter is 1cm, the column height is 15 cm), sequentially eluting with 50ml of water and 50ml of 65% ethanol, collecting the 65% ethanol eluate, filtering with a microporous membrane, and taking the subsequent filtrate to obtain the compound magnolia bark extract.

Preparation of a negative solution of Magnolia officinalis: according to the prescription (without magnolia officinalis) of the invention, a negative sample without magnolia officinalis is prepared by the process, about 0.8g is weighed, and the preparation method is the same as that of the compound magnolia officinalis extract test solution.

4: measurement method

Respectively and precisely sucking 10ul of cortex Magnolia officinalis decoction pieces solution, reference solution of reference medicinal material, reference solution of reference substance, compound cortex Magnolia officinalis extract sample solution, and cortex Magnolia officinalis negative solution, injecting into high performance liquid chromatograph, and determining according to specified chromatographic conditions.

5: methodology investigation

5.1: specialization inspection

The results show that: negative results have no interference on the detection of the magnolia officinalis, and the specificity is good. The results are shown in FIG. 4, where from top to bottom: data 1: a negative solution lacking magnolia bark; data 2: compound magnolia bark extract (batch No. 20220501); data 3: honokiol control solution, data 4: magnolol control solution.

5.2: precision degree

Taking a test solution of cortex Magnolia officinalis extract (batch No. 20220301), continuously injecting sample for 6 needles under the above chromatographic conditions, and recording chromatogram. The peak 9 (honokiol) was used as the S peak, and the relative retention time and RSD of the relative peak area of each characteristic peak and the S peak were calculated, and the results are shown in Table 4.

Table 4: results of precision investigation

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The results show that: the relative retention time RSD% and the relative peak area RSD% of each characteristic peak in the 6-needle test sample solution are both less than 2.0%, so the instrument precision of the method is good.

5.3: repeatability of

Taking compound cortex Magnolia officinalis extract (batch No. 20220301), preparing 6 parts of test solution, analyzing under determined chromatographic conditions, and recording chromatogram.

The peak 9 (honokiol) was used as the S peak, and the relative retention time of each characteristic peak and the S peak and the RSD of the relative peak area were calculated, and the results are shown in Table 5.

Table 5: results of repeated investigation

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The results show that: the relative retention time RSD% and the relative peak area RSD% of each characteristic peak in the 6-pin parallel samples are all less than 2.0%, which shows that the method has good repeatability.

5.5: stability of

Taking the compound cortex Magnolia officinalis extract (batch No. 220220301), standing at room temperature for 0h, 2h, 4h, 6h, 8h, 10h, 12h, 18h, and 24h, analyzing under determined chromatographic conditions, and recording chromatogram.

The peak 9 (honokiol) was used as the S peak, and the relative retention time and RSD of the relative peak area of each characteristic peak and the S peak were calculated, and the results are shown in Table 6.

Table 6: stability examination results

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The results show that: the relative retention time RSD% and the relative peak area RSD% of 16 characteristic peaks of the test solution are less than 2.0% within 24h, and the stability is good.

6: establishing characteristic map

6.1: selection of common peaks

Comparing the HPLC chromatogram of 15 batches of compound magnolia bark extracts with the reference magnolia bark medicinal materials and 15 batches of magnolia bark decoction pieces, finally selecting 16 peaks as common peaks (namely peaks 1-16) with large area, good separation degree and stable physicochemical properties, and obtaining the result as shown in figure 5, wherein peak 9 (S peak) is honokiol and peak 12 is magnolol.

6.2: calibration of common peaks and selection of reference peaks

Comparing the chromatogram of honokiol and magnolol reference substance with chromatogram peaks at corresponding positions in cortex Magnolia officinalis decoction pieces, cortex Magnolia officinalis reference medicinal material, and compound cortex Magnolia officinalis extract, it can be seen that the characteristic peak No. 9 in sample chromatogram is honokiol, and the characteristic peak No. 12 is magnolol. Since chromatographic peaks of honokiol in the chromatogram of each batch of samples are well separated, the peak area is large and is shared by all samples, the honokiol is determined as a reference peak (S peak), and the result is shown in figure 4.

6.3: evaluation of similarity of Magnolia bark extract

Introducing HPLC chromatograms of 15 batches of cortex magnoliae officinalis decoction pieces into software of 'traditional Chinese medicine chromatogram fingerprint similarity evaluation system' (2012.130723 version) of the State pharmacopoeia Committee, setting a reference spectrum, performing multi-point correction and peak matching on the chromatograms, and generating a reference characteristic spectrum of the 15 batches of cortex magnoliae officinalis decoction pieces by adopting an average method, wherein the result is shown in figure 5. Calculating the relative retention time of each characteristic peak and the S peak, wherein the relative retention time is as follows: 0.14 (peak 1), 0.24 (peak 2), 0.36 (peak 3), 0.44 (peak 4), 0.52 (peak 5), 0.59 (peak 6), 0.86 (peak 7), 0.97 (peak 8), 1.00 (peak 9), 1.02 (peak 10), 1.03 (peak 11), 1.05 (peak 12), 1.07 (peak 13), 1.08 (peak 14), 1.21 (peak 15), 1.33 (peak 16).

The 16 common chromatographic peaks in the test samples S1-S15 of the compound magnolia bark extract are subjected to multi-point correction and peak matching, and the similarity between 15 batches of the compound magnolia bark extract and magnolia bark decoction pieces is calculated, and the result is shown in Table 7 and figure 6.

Table 7: similarity calculation result of compound magnolia bark extract (15 batches in total) and magnolia bark decoction pieces

From the above, it can be seen that: the similarity of the 15 batches of compound magnolia bark extracts and the contrast characteristic spectrum of magnolia bark decoction pieces is higher and is more than 0.90. The chromatogram of the compound cortex Magnolia officinalis extract should present 16 chromatograms corresponding to the control characteristic chromatogram, wherein the retention time of peak 9 and peak 12 should correspond to the retention time of chromatographic peaks of honokiol and magnolol control. The characteristic spectrum detection method is stable and reliable, and can be used for quality control of the magnolia officinalis in the compound magnolia officinalis extract.

Comparative example 1

Weighing 52g of mangnolia officinalis, 33g of cherokee rose fruit meat, 18g of scorched hawthorn fruit, 26g of scorched malt, 35g of scorched medicated leaven, 102g of ginger and 96g of Chinese date meat (fresh), directly feeding the decoction pieces, adding water for decocting for 3 times, filtering, combining the filtrates, concentrating, drying under reduced pressure, crushing, and totally mixing to obtain the finished product.

Comparative example 2

Refer to thin layer identification method test under item "Cherokee Rose fruit" in the second part of 2020 edition of Chinese beast Pharmacopeia.

Preparation of a compound magnolia bark extract test solution: taking 2g of compound cortex Magnolia officinalis extract, adding 50ml of ethanol, performing ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, adding 20ml of water into residue to dissolve, extracting with ethyl acetate for 3 times, 30ml each time, mixing ethyl acetate solutions, evaporating to dryness in water bath, adding methanol into residue to dissolve and diluting to 2ml to obtain the final product.

Preparation of reference drug solution: weighing 2g of fructus Rosae Laevigatae reference medicinal material, and making into compound cortex Magnolia officinalis extract test solution.

The solution was pipetted up to 5ul, and spotted on the same silica gel G thin layer plate, developed with chloroform-ethyl acetate-methanol-formic acid (5.

And (4) conclusion: in the chromatogram of the test sample, there are no spots of the same color at the position corresponding to the chromatogram of fructus Rosae Laevigatae control drug, and the result is shown in FIG. 3, wherein 1-2 are fructus Rosae Laevigatae control drug, 3 is cortex Magnolia officinalis extract (batch No. 20220701), and 4 is cortex Magnolia officinalis extract (batch No. 20220702).

Test example 1

The contents of total sugar and total flavone in finished products of example 1 and comparative example 1 are respectively determined by the following method:

(1) Method for measuring total sugar

Preparation of control solutions: taking 70mg of anhydrous glucose dried to constant weight at 105 ℃, precisely weighing, placing in a 100ml measuring flask, adding water to dissolve, diluting to scale, and shaking up to obtain the product (0.7 mg of anhydrous glucose is contained in each 1 ml).

Preparation of a standard curve: precisely measuring the reference substance solutions 0.5ml, 1.0ml, 1.5ml, 2.0ml and 2.5ml, respectively placing in 50ml measuring bottles, adding water to the scale, and shaking. Precisely measuring 2ml of the solution, placing the solution in test tubes with plugs, precisely adding 1ml of 4% phenol solution respectively, mixing, rapidly and precisely adding 7ml of sulfuric acid, shaking, placing in a 40 ℃ water bath for 30min, taking out, placing in an ice water bath for 5min, taking out, measuring absorbance at 490nm by ultraviolet-visible spectrophotometry (general rule 0401) with corresponding reagents as blank, and drawing a standard curve with the absorbance as ordinate and the concentration as abscissa.

Preparing a test solution: taking about 0.2g of each of the finished products in example 1 and the finished products in comparative example 1, precisely weighing, placing in a conical flask with a plug, precisely adding 50ml of water, weighing, standing for 1 hour, heating and refluxing for 1 hour, cooling, weighing again, supplementing the lost weight with water, shaking up, filtering, precisely weighing 1ml of subsequent filtrate, placing in a 100ml measuring flask, adding water to the scale, shaking up, precisely weighing 25ml, placing in a 50ml measuring flask, adding water to the scale, shaking up, densely weighing 2ml, placing in a test tube with a plug, and measuring the absorbance according to the method under the preparation item of a standard curve from the point that 1ml of each precise 4% phenol solution is added.

The weight of total sugar (ug) in the test solution was read from the standard curve and the total sugar content in the sample was calculated.

(2) Method for measuring total flavonoids

Preparation of control solutions: taking an appropriate amount of rutin control, precisely weighing, and adding ethanol to obtain a solution containing rutin 0.15mg per 1 ml.

Preparation of a standard curve: precisely measuring 1ml, 2ml, 3ml, 4ml, 5ml and 6ml of reference substance solution, respectively placing the reference substance solution into 25ml measuring flasks, adding water to 6ml, adding 1ml of 5% sodium nitrite solution, uniformly mixing, standing for 6min, adding 1ml of 10% aluminum nitrate solution, uniformly shaking, standing for 6min, adding 10ml of sodium hydroxide test solution, adding water to the scale, uniformly shaking, standing for 15 min, taking a corresponding reagent as a blank, measuring absorbance at the wavelength of 500nm by an ultraviolet-visible spectrophotometry (general rule 0401), taking the absorbance as a vertical coordinate and the concentration as a horizontal coordinate, and drawing a standard curve.

Preparing a test solution: taking 0.10g of each of the samples in the examples and the samples in the comparative examples 1, precisely weighing, placing in a conical flask with a plug, precisely adding 25ml of dilute ethanol, sealing the plug, shaking up, carrying out ultrasonic treatment for 5min, placing for more than 3h, filtering, precisely weighing 2ml of subsequent filtrate, placing in a 25ml measuring flask, diluting to a scale with water, and shaking up to obtain a sample solution. Precisely measuring 2ml of a test sample solution, placing the test sample solution in a 25ml measuring flask, measuring the absorbance by a method from 'adding water to 6 ml', simultaneously precisely measuring 2ml of the test sample solution, placing the test sample solution in the 25ml measuring flask, adding water to the scale, and shaking up to obtain a blank solution according to the method under the preparation item of the standard curve.

And reading the rutin amount in the test solution from the standard curve, and calculating the content.

The results of the content measurement of the final product are shown in Table 8.

Table 8: comparison result of total sugar content and total flavone content in finished products of example 1 and comparative example 1

Numbering	Name of active ingredient	Comparative example 1	Example 1
				1	Total sugar content (%, g/g)	9.77％	43.83％
2	Total Flavonoids (%, g/g)	13.40％	41.26％

As can be seen from Table 8, the total sugar and total flavone of the active ingredients in example 1 are 43.83% and 41.26%, respectively, which are significantly higher than those of the active ingredients prepared by the conventional method (comparative example 1). The reason analysis is that the effective components are easier to dissolve out due to solid fermentation, and the macroporous adsorption resin can effectively remove ineffective impurities.

Test example 2

The male Kunming mice 72, weight 18-22g, were randomly divided into 6 groups of 12 mice each. Blank control group, model group, comparative example 1 group, and example 1 group (low, medium, and high dose groups), respectively. The administration was carried out by gavage, the dose of example 1 was 21.5mg, 43mg and 86mg/kg, the dose of comparative example 1 was 100mg/kg, and the administration was continued for 30 days by administering an equal volume of physiological saline to the blank control group and the model group. The mice in the model group were administered cyclophosphamide intraperitoneally at the 21 st day, and 5 times intraperitoneally every other day.

The MTT method is used for measuring the proliferation of splenic lymphocytes of mice, and the plantar thickening method is used for measuring the delayed type allergic reaction. Statistical analysis software SPSS22.0 is adopted for treatment, single-factor variance analysis is carried out, multiple comparisons are carried out among groups, differences are considered to have statistical significance when P is less than 0.05, and results are shown in tables 9-11.

Table 9: example 1, comparative example 1 Effect on the humoral immune function of mice

Group of	Dosage (mg/kg)	Number of antibody produced	Half maximal hemolysis value (HC) ₅₀ )
				Blank control group	/	141±23	104.42±7.64
Model set	100	76±31	77.10±8.03
				Comparative example 1 group	100	143±47	118.06±13.14
Example 1 Low dose group	21.5	105±47	99.06±17.09
				Dose groups of example 1	43	131±38	113.72±10.63
Example 1 high dose group	86	214±35*	135.14±11.07*

Note: p < 0.05 compared to control blank

Table 10: example 1, comparative example 1 Effect on immune function of mouse cells

Note: p < 0.05 compared to control blank

Table 11: example 1 and comparative example 1 Effect on the degree of swelling of toes in mice

Note: p < 0.05 compared to control blank

As can be seen from tables 9-11, the high dose group (86 mg/kg) in example 1 significantly increased the delayed type allergic reaction ability, lymphocyte transformation ability, antibody-producing cell number and half serum hemolysis value of mice compared to the blank control group, indicating that the compound Magnolia bark extract of the present invention can enhance the immunity of mice. Modern research suggests that lymphoid tissue of the gastrointestinal tract is an important component of peripheral lymphoid tissue with immune function, and digestive system diseases are mostly related to immune dysfunction.

Test example 3

60 Kunming mice with the weight of 18-22g are randomly divided into 5 groups, and each group has 12 mice. Blank control, model, example 1 (low, medium, high dose) groups were used. The preparation is administered by gastric gavage, the dosage of example 1 is 30mg, 60mg and 120mg/kg respectively, and the blank control group and the model group are administered with physiological saline with the same volume for 30 days continuously. The model group was modeled with compound diphenoxylate. After the experiment is finished, the test sample is fasted for 16 hours, the test sample is given once again on the day of measurement, the compound diphenoxylate is given to each experimental group and each model group after 30min, the distilled water is given to the blank control group, the indicator is given to each group after 30min, the animals are killed after 25min, the ink propulsion rate is calculated, and the measurement result is shown in the table 12.

Table 12: example 1 Effect on the rate of intestinal motility in mice

Group of	Dosage (mg/kg)	Ink push Rate (%)
			Blank control group	/	61.2±1.3
Model set	/	25.2±2.0
			Example 1 Low dose group	30	43.8±1.4#
Dose groups of example 1	60	52.1±2.4#
			Example 1 high dose group	120	57.8±1.7#

Note: compared with the model group, # P < 0.05

SD rats were 48, and were randomly divided into 4 groups of 12 rats each by body weight. Blank control group, example 1 group (low, medium, high dose group) respectively. The samples were administered by gavage, and the blank control group was given distilled water 1 time per day for 30 consecutive days. The dose of example 1 was 45mg, 90mg and 180mg/kg, respectively. Body weight and food intake were measured 2 times per week during the experiment and the dosage was adjusted according to body weight changes. Animal weight gain, feed intake and food utilization were calculated at the end of the experiment and the results are shown in table 13.

Table 13: example 1 Effect on weight gain, food intake and food availability in rats

Note: p < 0.05 compared to control blank

As can be seen from table 13, each of the dose groups of example 1 was able to modulate the propulsive effect of the small intestine as compared to the model group; as can be seen from Table 6, the dose group (90 mg/kg) and the high dose group (180 mg/kg) in example 1 increased the body weight, food intake and food utilization of rats, compared with the blank control group.

Test example 4

The inland river weaned piglets diagnosed with diarrhea were randomly divided into 2 groups of 20 piglets (half of the male parent) each, the administration dose of the group of example 1 was 110mg/kg once a day for three consecutive days, the blank control group was not treated, the number of piglets with diarrhea, the number of days with diarrhea, and other indices were measured, and the diarrhea rate and the diarrhea index were calculated, and the results are shown in table 14.

Table 14: EXAMPLE 1 therapeutic results on Anjiang post-weaning diarrhea

Note: p < 0.05 compared to control blank

As can be seen from table 14, the diarrhea rates of the inland river weaned piglets in the blank control group and the example 1 group are 79.2% and 6.67%, respectively, and the diarrhea indexes are 1.95 and 0.35, respectively. The diarrhea rate and the diarrhea index of the group of example 1 are both obviously lower than those of a blank control group (P < 0.05). The compound magnolia bark extract has obvious effect of treating diarrhea of weaned piglets in Nejiang river.

Test example 5

Selecting 60 sows with similar gestation times and healthy pregnancy time of 95-96 days, and randomly dividing the sows into 3 groups, namely an example 1 group, a comparative example 1 group and a blank control group, wherein each group comprises 20 sows. The feed is administered with water or mixed feed at a dose of 0.1g/kg body weight/day for 5 days. The time of the sow's labor was recorded, from the start of the sow's labor to the time of the last piglet's afterbirth discharge, and the results are shown in table 15.

Table 15: EXAMPLE 1 Effect on sow parturition results

Group of	Number of test heads (only)	Labor (min)
			Blank control group	20	289.93
Comparative example 1 group	20	267.38
			EXAMPLE 1 group	20	150.19*

Note: p < 0.05 compared to blank control and comparative example 1

As can be seen from Table 15, the parturient labor of the sows of example 1 was significantly reduced (P < 0.05) compared with the placebo group and the group of comparative example 1. The pregnant sow can benefit in the labor process when fed with the compound magnolia bark extract.

While the present invention has been described in detail with reference to the illustrated embodiments, it should not be construed as limited to the scope of the present patent. Various modifications and changes may be made by those skilled in the art without inventive work within the scope of the appended claims.

Claims

1. The compound magnolia bark extract is characterized by comprising the following raw materials in parts by weight:

1 to 60 parts of magnolia officinalis, 3 to 50 parts of cherokee rose fruit meat, 1 to 60 parts of scorched hawthorn fruit,

90-101 parts of jujube meat.

2. The compound magnolia bark extract as claimed in claim 1, wherein the compound magnolia bark extract comprises the following raw materials in parts by weight: 52-57 parts of mangnolia officinalis, 29-33 parts of cherokee rose fruit meat, 12-18 parts of scorched hawthorn fruit, 35-36 parts of scorched medicated leaven, 26-28 parts of scorched malt, 102-103 parts of ginger and 96 parts of Chinese date meat.

3. The preparation method of the compound magnolia bark extract is characterized by comprising the following steps:

step (1): pulverizing charred Massa Medicata Fermentata into fine powder, pulverizing cortex Magnolia officinalis, fructus Rosae Laevigatae, charred fructus crataegi and charred fructus Hordei Germinatus into raw material micropowder;

4. The method for preparing the compound magnolia bark extract as claimed in claim 3, wherein: the composite probiotic powder in the step (4) comprises 1 part by mass of monascus anka, 0.8-3.6 parts by mass of lactobacillus plantarum and 1-1.7 parts by mass of bifidobacterium bifidum, and the mass ratio of the inoculation amount of the composite probiotic powder to the fermentation substrate is 1:46-130.

5. The method of claim 3, wherein the method comprises the steps of: stirring and extracting the fermentation raw materials in the step (5) for three times, wherein the additive added in the first stirring and extracting is a mixture of a medium and enzyme, the medium is lactic acid, hydrochloric acid, citric acid and water, the enzyme is pepsin, chymosin and lipase, and the extracting time is 0.5h; the additive added in the second stirring extraction is a mixture of a medium and enzyme, wherein the medium is sodium bicarbonate and water, the enzyme is lactase, amylase and trypsin, and the extraction time is 1h; the additive added in the third stirring extraction is a medium, wherein the medium is sodium carbonate and water, and the extraction time is 2h.

6. A quality control method of a compound magnolia bark extract is characterized in that: comprises a detection method of cherokee rose flesh and a detection method of a magnolia officinalis characteristic spectrum.

7. The quality control method of magnolia bark extract as set forth in claim 6, wherein the detection method of cherokee rose fruit comprises the following steps:

step a: preparing a test solution: grinding 1g of the product, extracting with solvent, filtering, evaporating filtrate, dissolving residue with solvent, and diluting to obtain sample solution;

step c: and (3) determination: sucking the sample solution and the reference solution, respectively spotting on the same silica gel G thin layer plate, developing in developing agent, taking out, air drying, and inspecting.

8. The quality control method of magnolia bark extract as claimed in claim 7, wherein: the extraction solvent in the step a is ethyl acetate, and the extraction mode is ultrasonic extraction; the extraction solvent in the step b is ethyl acetate, and the extraction mode is any one of ultrasonic extraction and heating reflux extraction; the developing agent in the step c is a mixture of petroleum ether, ethyl acetate and formic acid, wherein the volume ratio of the petroleum ether to the ethyl acetate to the formic acid is 100.

9. The quality control method of magnolia bark extract as claimed in claim 6, wherein the detection method of the magnolia bark characteristic spectrum comprises the following steps:

step a: preparation of reference solutions: weighing honokiol reference substance, and adding solvent to obtain reference substance solution;

10. The method for controlling the quality of magnolia bark extract as claimed in claim 9, wherein the chromatographic conditions in step c are: the chromatographic column is C18, the filler is octadecylsilane chemically bonded silica, the mobile phase is a mixed solution of methanol and 0.4% formic acid, wherein the methanol is a mobile phase A, the 0.4% formic acid solution is a mobile phase B, the flow rate is 1ml/min, the column temperature is 30 ℃, and the detection wavelength is 230nm.