CN107233366A - A kind of leaf of Moringa granule and preparation method and application - Google Patents
A kind of leaf of Moringa granule and preparation method and application Download PDFInfo
- Publication number
- CN107233366A CN107233366A CN201710348607.2A CN201710348607A CN107233366A CN 107233366 A CN107233366 A CN 107233366A CN 201710348607 A CN201710348607 A CN 201710348607A CN 107233366 A CN107233366 A CN 107233366A
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- moringa
- leaf
- granule
- preparation
- liver
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1617—Organic compounds, e.g. phospholipids, fats
- A61K9/1623—Sugars or sugar alcohols, e.g. lactose; Derivatives thereof; Homeopathic globules
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention belongs to biological medicine and field of food, a kind of preparation method of leaf of Moringa granule is disclosed.Using leaf of Moringa as primary raw material, Icing Sugar and mannitol are auxiliary material.Its core is that leaf of Moringa pulverizes and sieves, and refluxing extraction is carried out for several times by solvent of distilled water, merges extract solution；Aqueous extracts are purified by ethanol precipitation centrifugation, obtained refined solution is concentrated under reduced pressure, dried again, obtains leaf of Moringa dry extract；Then dry extract is equipped with auxiliary material, granule is made using the method for wet granulation.Compared with prior art, leaf of Moringa granule of the invention makes the effective active composition in leaf of Moringa highly concentrated, stable and controllable for quality, palatability and compliance are good, it is easy to take, is adapted to industrialized production, and there is provided its application in preventing and treating immunological liver injury.
Description
Technical field
The invention belongs to biological medicine and field of food, more particularly, to a kind of leaf of Moringa granule and its preparation side
Method and application.
Background technology
Immunological liver injury (Immunological Liver Disease, ILD) is by various exogenous or autoantigen
Sexual factor induces the disorder of Liver immunity responsing reaction, so that the disease of damage lesion.Immunological liver injury pathogenesis and body
Mechanism of immunotolerance is destroyed, and the lethal cell recognitions of KC are obscured, and nospecific immunity cell largely infiltrates, and inflammatory factor release is drawn
Hepatotoxicity and damage are played, if not in time and obtaining appropriate treatment, fibrosis, the hardening of liver is often evolved to, in addition it is swollen
Knurl.In terms of risk factor, immunological liver injury Etiological has hepatites virus infections, autoimmunity exception, after liver transplantation, thermophilic
Wine, Irrational Use of Drugs etc., wherein, it is very big with the immunological liver injury accounting that hepatites virus infections trigger.China is viral liver
Scorching high Prevalent district, shows according to the statistics of in August, 2016, without Hong Kong, Macao and Taiwan, and China's hepatites virus infections morbidity is up to 123,
297, wherein hepatitis B is fallen ill 97,670, and hepatitis is fallen ill 19,250.Therefore, the immunological liver that hepatites virus infections trigger is damaged
The preventing and treating of wound is when the severe task of previous item.Also it can not be ignored using caused immunological liver injury in addition, medicine is unreasonable,
Crowd such as special constitution is easy to trigger in liver the hypersensitivity of immune response after some medicines are taken, to liver
Cause different degrees of infringement.Research is thought at present, can trigger the medicine of immunological liver injury more than 1,000 kind (David
S,Hamilton JP.Drug-induced liver injury.Nat Rev Gastroenterol Hepatol,2010,1
(6):73-80).Therefore, medicine and functional food that searching and research and development can effectively prevent and treat immunological liver injury are very necessary.
At present, the remedy measures of immunological liver injury are to find early and integrated control, and therapeutic strategy stops including antiviral
Liver cytotoxic drug, scavenging capacity oxygen radical, inflammatory mediator, immune cell factor etc. are used, to liver cell mitochondria, cell membrane flow
Property, cytoskeleton carry out restoration and protection, improve Na+-K+- atpase activity etc..Using suppression or elimination are lured for a long time, to greatest extent
The various factors of immunological liver injury is sent out, regulation immune response recovers normal, so as to delay or prevent disease development.Always
It, immunological liver injury needs comprehensive, chronicity treatment.In this regard, exploitation one kind can play comprehensive liver protection effect, poison is secondary to be made
With small, body drug-resistant intensity is high, can long-term taking auxiliary therapeutical agent it is very necessary.In research, it has been found that leaf of Moringa particle
Agent has preferable protective effect to the experiment mice immunological liver injury that BCG combined LPS is induced, and can be used as immunological liver
The auxiliary treatment purposes of damage.
Moringa (Moringa oleifera Lam.), is Moringa suborder Moringaceae (Moringaceae) Moringa
(Moringa) plant, is perennial tropical and subtropical zone deciduous tree, and Yin Qigen has acid, therefore Moringa of thus gaining the name.Moringa
Root, leaf, the equal edible of okra fruit, containing several mineral materials, vitamin, rich in nutrition content；In addition, its root, skin, leaf, fruit are all
Medical material can be made, therefore, Moringa is described as " tree of miracle ", " diamond in plant ".Moringa has a variety of medical values, existing
Generation research thinks, it can with control ulcer, hypertension, high fat of blood, cancer, diabetes, inflammation, bacterium and virus infection etc. disease
Disease, is the plant of great research and development potentiality.Foreign countries have been related to the research report of Moringa liver protection at present, and Sharifudin etc. is used
Paracetamol (APAP) inducing mouse hepatic injury, using N-acetylcystein as control drug, have studied leaf of Moringa and flower
Therapeutic action of the extract to mouse liver injury, as a result shows the mouse medicine that leaf of Moringa and flower extract are excessively caused to APAP
Property hepatic injury has therapeutic action (Sharifudin SA, Fakurazi S, Hidayat MT, et al.Therapeutic
potential of Moringa oleifera extracts against acetaminophen-induced
Hepatotoxicity in rats.Pharmaceutical Biology, 2013,51 (3)：279-288).Sharma etc. is ground
The protective effect for the acetsminophen that Moringa is induced 7,12- dimethylbiphenyls [a] anthracene (DMBA) is studied carefully, it is found that it can drop
Liver aspartate transaminase (AST), alanine aminotransferase (ALT) and the alkaline phosphatase of the liver injury model mouse of low DMBA inductions
Enzyme (ALP) level, significantly reverses the pathological change of the liver organization of DMBA inductions, it is believed that it is to hepatocellular injury caused by DMBA
With good liver protection potentiality (Sharma V, Paliwal R, Janmeda P, et al.Chemopreventive
Efficacy ofMoringa oleifera pods against 7,12-dimethylbenz [a] anthracene
induced hepatic carcinogenesis in mice.Asian Pacific Journal ofCancer
Prevention, 2012,13 (6)：2563-2569).Sinha etc. have studied moringa oleifera leaf extractive causes liver to radiation
The influence of dirty lipid peroxidation injury.Mouse after being administered 15 days is radiated exposed to 60Co- γ, observation irradiation model mice
The indexs such as liver lipid peroxidation, reduced glutathione, it is believed that moringa oleifera leaf extractive can prevent hepatic lipid peroxidation to damage
(SinhaM, Das DK, Datta S, et al.Amelioration ofionizing radiation induced lipid
peroxidation in mouse liver by Moringa oleifera Lam.leaf extract.Indian
Journal ofExperimental Biology, 2012,50 (3)：209-215).But, in terms of current research report, only
Research on leaf of Moringa to hepatotoxic compound chemical damage as caused by DMBA and APAP, and have no moringa oleifera leaf extractive
To the Effect study of liver immune response immunological liver injury caused by disorderly.
In addition, from the point of view of current production technology and the state of the art, to the use of leaf of Moringa mainly using it is soaked, decoct,
The method such as tabletting after being milled (fecula) or being milled.For soaked and decoction, be not suitable for industrialized production；Fecula is taken, human body
Bioavilability is low, and active ingredient absorbs few, is difficult to swallow, it is difficult to work；Fecula tabletting, the daily dose of user is from every
It control amount is far apart, and quality standard is difficult to control to, and does not reach effect of health care.
The content of the invention
The invention aims to the defect for overcoming prior art, there is provided a kind of preparation method of leaf of Moringa granule.
It is a further object of the present invention to provide leaf of Moringa granule prepared by a kind of above method.
Another object of the present invention is to provide the application of above-mentioned leaf of Moringa granule.
Above-mentioned purpose of the present invention is achieved by the following technical programs：
A kind of preparation method of leaf of Moringa granule, is comprised the following specific steps that：
S1. get the raw materials ready：Leaf of Moringa is dried, crush and sieved, leaf of Moringa powder is obtained；
S2. extract：Add water in leaf of Moringa powder, extracted at 60~100 DEG C, obtain the Aqueous extracts of leaf of Moringa；
S3. purify：The Aqueous extracts of leaf of Moringa are added in ethanol and precipitated, supernatant, as Moringa are taken after centrifugation
Leaf refined solution；
S4. concentrate：Leaf of Moringa refined solution is concentrated under reduced pressure under the conditions of 60~80 DEG C, leaf of Moringa thick paste is obtained；
S5. dry：Leaf of Moringa thick paste is dried in vacuo at 60~80 DEG C, leaf of Moringa dry extract is obtained；
S6. pelletize：By leaf of Moringa dry extract and auxiliary materials and mixing, leaf of Moringa granule is obtained using wet granulation.
Preferably, the mesh number sieved described in step S1 is 80~100 mesh.
Preferably, the mass ratio of leaf of Moringa powder and water described in step S2 is 1：(15~25)；The time of the extraction is
0.5~1.5h；The number of times of the extraction is 1~3 time.
Preferably, the volume ratio of the Aqueous extracts of leaf of Moringa and ethanol described in step S3 is 1:(1~3)；The ethanol
Concentration is 60%~80%；The rotating speed of the centrifugation is 8000~14000rpm；The time of the centrifugation is 10~20min.
Preferably, the vacuum depressurized described in step S4 is -0.06~0.07MPa；Institute is vacuum drying in step S5
Vacuum is -0.06~0.07MPa.
Preferably, the relative density of leaf of Moringa thick paste described in step S4 is 1.30~1.35.
Preferably, auxiliary material described in step S6 is Icing Sugar and mannitol.
It is further preferable that the mass ratio of the Icing Sugar and mannitol is (1~5):(5~1).
Preferably, the mass ratio of leaf of Moringa dry extract and auxiliary material described in step S6 is (1~3):(3~5).
Leaf of Moringa granule and its application in preventing and treating immunological liver injury prepared by the above method.
The leaf of Moringa granule of the present invention combines lipopolysaccharides to BCG vaccine (Bacillus Calmette-Gu é rin, BCG)
Reflect the paddy third of hepatocellular injury in the Immune liver injury mice serum of (Lipopolysaccharide, LPS) induction
Transaminase (ALT) and glutamic-oxalacetic transaminease (AST) are significantly reduced effect, and to reflecting the superoxides of hepatic antioxidative function
Mutase (SOD) and glutathione peroxidase (GSH-Px) vigor have humidification, to reflection liver lipid peroxide water
Flat MDA (MDA) has reduction effect, is conducive to the defence and the reparation to liver of immunological liver injury, makes hepatocyte function
Damage mitigates or recovered normal.
ALT, AST in serum are mainly derived from liver cell, participate in synthesis and the catabolism of amino acid.Work as liver cell
When being caused the pathology damages such as inflammation, necrosis by various influences, the rupture of cell membrane can cause a large amount of of ALT, AST to flow into outside
Enter body circulation, ALT, AST level in blood is significantly raised, be present in after centrifugal blood in serum.Therefore, serum transaminase is
Reflect the important indicator of hepatocellular injury in clinical Liver function grade.
SOD is the primary protein substance of the various oxygen radicals of removing existed in body, to SOD water in hepatic homogenate
Flat detection can reflect the oxidation resistance of liver；GSH-Px is a kind of important catalyzing hydrogen peroxide being widely present in body
(H2O2) decompose enzyme.It is specifically catalyzed reduced glutathione (Glutathione, GSH) to H2O2Reduction reaction, can
To play protection membrane structure and fully functional effect.GSH-Px can promote H2O2With GSH reaction generations H2O and oxidation
Type glutathione (GSSG), GSH-Px vigor can use the consumption of GSH in enzymatic reaction to deduct non-enzymatic reaction (redox
Reaction) caused by GSH consumption represent.Therefore the detection to GSH in liver homogenate can reflect the oxidation resistance of liver；
MDA is one of most important product of membrane lipid peroxidatio, and its generation can cause the crosslinking of the large biological molecules such as protein, nucleic acid
Polymerization, makes function damage or the forfeiture of film, produces cytotoxicity, and the detection to MDA levels in hepatic homogenate can also reflect
The oxidation resistance or liver peroxide injury degree of liver.
Compared with prior art, the invention has the advantages that：
1. the preparation method of the present invention can make in leaf of Moringa by the water extraction of optimization, purifying, concentration, dry and moulding process
Effective active composition it is highly concentrated, quality controllable stabilization, palatability and compliance are good, are easy to take, be adapted to industrialized production.
2. leaf of Moringa granule of the present invention has humidification to liver SOD and GSH-px enzyme activity, it is more to combine fat to BCG vaccine
The Immune liver injury experiment mice of sugar induction shows obvious liver protection.
Brief description of the drawings
Fig. 1 is the standard curve of rutin.
Embodiment
Present disclosure is further illustrated with reference to specific embodiment, but be should not be construed as limiting the invention.
Unless otherwise specified, the conventional meanses that technological means used in embodiment is well known to those skilled in the art.Except non-specifically
Illustrate, the reagent of the invention used, method and apparatus is the art conventional reagent, methods and apparatus.
The DDB described in embodiment (Biphenyldicarboxylate, BP) dripping pill is a Community in Baiyunshan, Guangzhou group of stars
(medicine company) limited company produces；ALT, AST, MDA, SOD, GSH-Px enzyme detection kit builds up biological work by Nanjing
Journey research institute provides.
The preparation of the leaf of Moringa granule of embodiment 1
1. get the raw materials ready：Leaf of Moringa dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
2. extract：The leaf of Moringa powder obtained in step 1 is taken, using water as solvent, solid-liquid ratio is 1:20, extracted at 60 DEG C
1h, extraction time is 3 times, merges to obtain leaf of Moringa Aqueous extracts；
3. purifying：The leaf of Moringa Aqueous extracts obtained in step 2 are taken, the ethanol precipitation of equivalent 80% is added, turned in 12000rmp
The lower centrifugation 10min of speed, takes supernatant；
4. concentration：The leaf of Moringa refined solution obtained in step 3 is taken, be concentrated under reduced pressure under the conditions of 80 DEG C (vacuum -0.06~
0.07Mpa) to relative density 1.30~1.35, leaf of Moringa thick paste is obtained；
5. dry：The leaf of Moringa thick paste obtained in step 4 is taken, in being dried at 70 DEG C (vacuum -0.06~
0.07Mpa), leaf of Moringa dry extract (moisture ﹤ 3%) is obtained；
6. granulation：The leaf of Moringa dry extract obtained in step 5 is taken, ormal weight extract powder and auxiliary material are taken by the prescription 1 of table 1
(1:3) mix, leaf of Moringa granule is obtained using wet granulation.
The prescription compatibility of medicines of the different auxiliary material of table 1 and extract powder
Preparation process condition realizes optimization, is obtained by orthogonal experiments, and experimental method and data are referring to implementation
Example 2-6 concrete mode.
The preparation of the leaf of Moringa Aqueous extracts of embodiment 2
Embodiment high spot reviews Extracting temperature A, extraction time B, solid-liquid ratio C, tetra- factors of extraction time D, each factor
3 levels are taken, using L9(3)4Orthogonal arrage arrangement is tested, and screens optimum process condition, and factor level is shown in Table 2.Flavones is leaf of Moringa
Main composition, the leaching effect of the composition represents the leaching situation of active ingredient, and its assay has maturation method,
It is evaluation index (the higher the better for recovery rate) to select it, and weight coefficient is 0.6；Dry extract yield is used as the direct of impurity-eliminating effect simultaneously
Characterize, it is evaluation index (dry extract is more low better) to select it, and weight coefficient is 0.4.According to water extraction experiment of single factor, finally enter
L is gone9(3)4The experiment that orthogonal arrage is arranged, it is specific as shown in table 2.
Water extraction orthogonal arrage experimental design：According to the experiment of single factor of water extraction, it is determined that investigate Extracting temperature A, extraction time B,
Solid-liquid ratio C, tetra- factors of extraction time D, design L9(3)4It is orthogonal as shown in table 2.
The quadrature factor water-glass of table 2
Dry extract is measured according to orthogonal test table 2, extract solution is concentrated into below 50mL, 1 is carried out with petroleum ether：1
Extraction, stands after after complete layering, removes a layer sample liquid constant volume 50mL, respectively take 40mL sample liquids in 75mL evaporating dishes, be placed in
It is evaporated to dryness in baking oven, in being weighed on electronic balance, as shown in table 3, remaining sample liquid keeps sample preservation record data.
The measurement result of the dry extract of table 3
Set up the standard curve of rutin：
0.0100g rutin standard items accurately are weighed, is dissolved in 50ml distilled water, is placed in brown bottle, are carried out with ultrasound
Dissolving, as 0.1mg/mL rutin standard liquids.Respectively take 0.0,1.0,2.0,3.0,4.0,5.0mL titers in test tube, respectively
Plus distilled water is to 5mL；After add 5%NaNO2Solution 0.5mL, stands 5min, 10%AlCl3Solution 0.5ml, stands 5min, 4%
NaOH solution 4mL, stands 10min, and the absorbance value of rutin standard liquid is determined under 510nm wavelength.
The experimental data of the rutin standard curve of table 4
By the measurement result of table 4, cast out improper data (0.8,0.729), and thus do scatter diagram, linear fit is obtained：y
=0.8753x+0.0059, R2=0.9983, draw rutin standard curve as shown in Figure 1.
Orthogonal sample determination of total flavonoids：Sample liquid in table 2 respectively takes 0.1mL in test tube, plus distilled water is to 5mL,
Similarly add 5%NaNO2Solution 0.5mL, stands 5min, 10%AlCl3Solution 0.5ml, stands 5min, 4%NaOH solution 4mL,
10min is stood, the absorbance value of each sample liquid is determined under 510nm wavelength, and contained by standard curve calculating general flavone percentage
Amount, its result is as shown in table 5.It can show that comprehensive grading 41.667% is highest under the extracting factor of tested number 8 from table 5.
According to weight distribution, calculated just according to general flavone content (accounting for weight 0.6) and dry extract yield (accounting for weight 0.4)
Hand over the comprehensive grading of experiment, it is alternatively that one of basis of optimal experimental program, the standard of interpretation of result is according to formula (1) Suo Shi.
The orthogonal experiments of the leaf of Moringa aqueous extract of table 5 and its analysis
The orthogonal analysis of variance table of the leaf of Moringa aqueous extract of table 6
The results of analysis of variance in table 6, each factor influence power order be：D extraction times>B extraction times>C
Solid-liquid ratio>A Extracting temperatures.On the other hand, from the analysis of table 5, optimal extracting factor is：60 DEG C of Extracting temperature, is carried
Take 1h, solid-liquid ratio 1:20, extract 3 times.
The purifying process of the Aqueous extracts of embodiment 3
In order to retain active ingredient to greatest extent, the effect of removal of impurities is reached again, and Aqueous extracts carry out pure using centrifugation technique
Change is handled, and compares research to different centrifugal speeds.In view of the rotating speed limitation of actual centrifugation apparatus, setting speed
8000th, 10000 and 12000rpm is compared, and quadrature factor water-glass is as shown in table 7.
The quadrature factor level of the purifying process of table 7
2.044g leaf of Moringa powder is weighed, takes optimum extraction scheme to be extracted.After take 20 2mL centrifuge tubes, use liquid relief
Rifle pipette samples liquid 1mL and corresponding concentration ethanol 1mL, centrifugal purification is carried out according to the quadrature factor water-glass 7 of purifying process, after
2mL centrifugates are taken in colorimetric cylinder, same operation carries out 510nm light absorption value measure, determine and orthogonal the results are shown in Table 8.
With liquid-transfering gun 1:1 accurate pipette samples liquid and 80% ethanol, is well mixed, 10min is centrifuged under 12000rpm, returns
Centrifugate is received, 50mL extract solutions is separately taken, while being dried, weighs dry cream quality；Other extracting centrifugal liquid 2mL, at the same take not from
Heart liquid 1mL, adds 80% ethanol 1mL, plus distilled water is to 5mL, shakes up, and adds 5%NaNO2Solution 0.5mL, stands 5min,
10%AlCl3Solution 0.5mL, stands 5min, 4%NaOH solution 4mL, stands 10min, is determined under 510nm wavelength and absorbs light
Angle value, compares flavones content.Meanwhile, rutin standard curve drafting is similarly carried out, is learnt by linear fit result：Y=
1.0610X, R2=0.9998.
The purifying process quadrature factor of table 8 is analyzed
Learnt by the analysis result of table 8, purifying optimum process condition is：Using 80% ethanol 1:1 mixing, 12000rpm,
Centrifuge 10min.
The concentration technology of the Aqueous extracts of embodiment 4
Leaf of Moringa is extracted by Optimized Extraction Process condition, Aqueous extracts filtration, merging filtrate, centrifugal purification takes supernatant, pressed
Volume is divided into 3 equal portions, is concentrated under reduced pressure (vacuum -0.06~0.07Mpa) under the conditions of 60 DEG C, 70 DEG C and 80 DEG C respectively to relative
The thick paste of density 1.30~1.35, determines the content of general flavone, as shown in table 9.
(60 DEG C -1 is extracted by optimal case:20-1h-3 times), purifying (80% ethanol 1:1 mixing, 12000r, centrifugation
10min), the rear 45mL purifying sample liquid that accurately measures every time is concentrated, and temperature is controlled at 50 DEG C, 60 DEG C, 70 DEG C and 80 DEG C respectively,
Each concentration time is recorded, concentrate is concentrated into below 10mL, and constant volume is 10mL.Concentrate is taken to carry out the survey of general flavone content
It is fixed.
The screening of the concentration technology of table 9 and its test result
Learnt by table 9, when concentration time is 80 DEG C, flavones content and polyoses content are relatively most, and efficiency value is maximum,
It is thus determined that optimal thickening temperature is 80 DEG C.
The screening of the drying process of embodiment 5
Leaf of Moringa is extracted by Optimized extraction techniques, Aqueous extracts are filtered, and merging filtrate, centrifugal purification takes supernatant, by volume
Classify in three categories part, and decompression (vacuum -0.06~0.07Mpa) is concentrated into the thick paste of relative density 1.30~1.35, respectively at 50
DEG C, 60 DEG C, be dried (vacuum -0.06~0.07Mpa) at 70 DEG C and 80 DEG C, dry cream moisture ﹤ 3%.Dry cream is crushed, standby
With, and determine the content of general flavone and polysaccharide.
By optimal concentration technology, centrifugate is concentrated, 10mL is respectively taken every time, drying temperature be set as 50 DEG C, 60 DEG C,
70 DEG C, 80 DEG C, the dry time is recorded, completion is dried and takes specific phase homogenous quantities dry cream to dissolve, 10mL distilled water is added, ultrasound is molten
Solution to liquid is clarified, and carries out the assay of general flavone.
The selection result of the drying process of table 10
As shown in Table 10,70 DEG C when general flavone content preserve it is more intact, excessive temperature may cause flavones knot
The destruction of structure, crosses low temperature because drying time length also has certain influence to the content of general flavone, polyoses content influence is little, because
This determination optimum drying temperature is 70 DEG C.
The technique of the preparations shaping of embodiment 6
Pelletized using wet granulation technology.The product of wet granulation have good appearance, good fluidity, wearability compared with
By force, the advantages of compressibility is good.The hygroscopicity of particle, mouldability, melting, heap are mixed evenly to prepare according to auxiliary material and extract powder
Density and angle of repose select appropriate auxiliary material.
After recipe quantity medicinal material is taken by the optimum extraction process extraction, removal of impurities, concentration preferably gone out, dry dry extract is pressed
The prescription of table 1 takes ormal weight extract powder and auxiliary material (1:3) mix, determine the mouldability of leaf of Moringa granule, heap density, angle of repose with
And hygroscopicity, and using mouldability, heap density, angle of repose, melting and hygroscopicity as index comprehensive evaluation, it is preferably optimal auxiliary
Material, the standard as a result tested is carried out according to formula (2) Suo Shi.
The prescription compatibility of medicines design of the different auxiliary material of table 11 and extract powder
The measure of 6.1 mouldabilities
The particle prepared is weighed, a sieve (10 mesh sieve) is first crossed, sieves (65 mesh sieve) after No. four, collection can pass through one
Number sieve but can not by No. four sieve particles, weigh.Mouldability is calculated according to formula (3) and (4), and its measurement result is shown in Table 12.
The measure of the mouldability of the leaf of Moringa granule of table 12
The measure of 6.2 heap density
Heap density is also known as apparent density or bulk density, means the quality of unit volume particle.Volume used in heap density is
Refer to the cumulative volume including space between particle and itself space and particle and particle.The heap density of particle is heap volume greatly
It is small, can represent the close degree of particle and determine volume divided dose number.Particle after sieving is put into dry graduated cylinder
In, gently vibrate, read the mL numbers (V) at its nearly scale；Using the quality after sieving as M (g), both ratio is that heap is close
Degree.The calculating of heap density score value is carried out according to formula (5), and it the results are shown in Table 13.
The measure of the heap density of the leaf of Moringa granule of table 13
The measure at 6.3 angles of repose
Mobility is one of critical nature of granule, the quality of mobility and quality, the degree of accuracy of divided dose of particle
It is relevant, commonly use turnover rate in pharmacy and angle of repose is represented.Angle of repose is smaller, and mobility is better.Usual particle diameter is smaller or granularity
The wide particle of distribution, its body angle till is bigger；And particle diameter circle, big and uniform particle easily flow, angle of repose is small.
Using fixed funnel method, 3 funnels are connected and are fixed on the graph paper of horizontal positioned at 1cm height, it is small
Heart is poured into particle along hopper walls in the funnel on most until the particle cone tips formed on graph paper touch funnel
Mouthful untill, by graph paper measure conical base diameter ((2R), calculates angle of repose tga=H/R, does 3 times, calculate average value.
Angle of repose score value is calculated according to formula (6), and its result is as shown in table 14.
Wherein,
The measure at the angle of repose of the leaf of Moringa granule of table 14
6.4 hygroscopic measure
A certain amount of granule is taken, constant weight in 40 DEG C of baking ovens is put, bottom is placed with to the glass desicator of NaCl saturated solutions
Middle timing is put into NaCl until forming NaCl supersaturated solutions, and now the relative humidity in drier is 75%.In constant weight
Flat measuring cup bottom, which is put into after 2mm granule, correct amount, to be placed in above-mentioned drier (flat measuring cup opening).Claim after 48h
Amount, calculates moisture absorption percentage.Two groups are done altogether, calculate average value.Hygroscopicity is calculated according to formula (7) and (8), its test result
As shown in Table 15.
The hygroscopicity of the leaf of Moringa granule of table 15 is determined
The overall merit of preparations shaping technique is according to formula (2), and its result is as shown in table 16.As can be known from Table 16, it is optimal
Ratio of adjuvant scheme is formula number 1 (pure extract powder 2g, Icing Sugar 1g, mannitol 5g).
The overall merit of the leaf of Moringa granular preparation moulding process of table 16
The leaf of Moringa granule of embodiment 7 combines the immunological liver injury mould caused by lipopolysaccharides (LPS) to BCG vaccine (BCG)
The influence of type mouse
1. material：
(1) experimental animal：84 SPF grades of Kunming (KM) are planted into male mice and are randomly divided into 6 groups, respectively blank control
Group, model group, DDB (15mg/mL) group, the high, medium and low dosage of leaf of Moringa granule of the present invention (concentration be respectively 2g/mL,
1g/mL, 0.5g/mL) group, every group of each 14 mouse.The SPF grades of Kunming (KM) plants 18~22 grams of the weight of male mice,
Experimental animal credit number：SCXK (Guangdong) 2013-0034, by Traditional Chinese Medicine University Of Guangzhou, Experimental Animal Center is provided.Use license
Card number：SYXK (Guangdong) 2014-0136.
(2) reagent：The leaf of Moringa dry cream that 1-6 of the embodiment of the present invention is prepared；Bifendate, by Guangzhou white clouds
A mountain group of stars (medicine company) limited company produces；AST, ALT, MDA, SOD and GSH-PX detection kit build up biology by Nanjing
Graduate School of Engineering is provided.
2. mouse adaptability is raised and packet
Experiment mice 84, adapts to environment and raises one week, and only, drinking water is enough, and replacing in two days is once padded by daily feeding 4g/
Material.After one week, mouse is randomly divided into 6 groups (every group 14) according to body weight：Blank control group, model group, Moringa Leave extract 3
Individual dosage group (2mg/kg, 1amg/kg, 0.5amg/kg), DDB positive controls (150mg/kg).
3. test method：Immunological liver injury model is set up using BCG vaccine (BCG) stuffing polysaccharide (LPS).Experiment
Preceding adaptability is raised 7 days, free diet, the food ration and amount of drinking water of daily observed and recorded each group mouse.Experiment first day, except sky
Outside white control group, remaining mouse is through tail vein injection 0.25mg (0.2mL, containing about 2.5 × 105Individual viable count) BCG.After 3h, remove
Blank control group and model group mouse stomach are given outside normal saline, each dose of test group mouse give daily 2 times it is tested
Thing, by 0.12mL/10g weight gavages, DDB group mouse presses the dosage medication after the daily quantity conversion of people, i.e.,
150mg/kgBW dosage gavages, one time a day.Record each group mouse weight is weighed within every 2 days, to adjust corresponding tested material consumption.
Test the 10th day, remaining mouse after last time administration 3h in addition to blank control group is through the μ g (0.2mL) of tail vein injection 7.5
LPS, fasting (can't help water) 16h, weighs body weight, then plucks eyeball and take blood, and the whole blood of collection is placed after 1h at 20 DEG C, through turning
Fast 3000rpm centrifuges 10min, draws serum in sterilizing 1.5mL centrifuge tubes, is placed in -20 DEG C of refrigerators and saves backup.Cervical dislocation
Mouse is put to death, mice organs is weighed, liver is placed on ice platform, liver lobus dexter is quickly cut with knife blade, notes taking liver right
Leaf same area hepatic tissue, which is divided into 2~3 parts and is distributed into clean cryopreservation tube and is placed in -80 DEG C of refrigerators, to be saved backup.
4. Testing index and detection method：Glutamic-oxalacetic transaminease (AST) activity and glutamic-pyruvic transaminase (ALT) activity in serum
Determine, carried out in strict accordance with kit specification operating procedure.MDA (MDA) content, superoxide dismutase in hepatic tissue
(SOD) measure of activity and glutathione thing peroxidase (GSH-px) activity.A liver organization of above-mentioned preservation is taken, with cold
Normal saline is operated into 10% liver homogenate by kit specification operating procedure, and MDA contents, SOD live in detection liver
Property, GSH-px activity.
5. statistical analysis：Experimental data carries out statistical disposition using SPSS 17.0 software kits, as a result with Table
Show, comparing between group and using one-way analysis of variance, P ＜ 0.05 indicate significant difference.Eyeball takes blood, separates serum, determines
MDA and triglycerides (TG) content in mice serum；Liver organization is taken to detect the vigor of MDA, SOD enzyme activity and GSH-px enzymes,
Its result is as shown in table 17 and table 18.
6. result of the test：
Influence of the leaf of Moringa granule of table 17 to immunological liver injury in mice Serum ALT and AST activity
Note：Compared with model group, * represents P<0.05, * * represents P<0.01.Compared with blank group, # represents P<0.05, ##
Represent P<0.01.
Drawn, compared with blank control group mouse by table 17, the activity of ALT and AST enzymes is obvious in model group mice serum
Rise, with significant significant difference (P<0.01), illustrate that BCG adds LPS inducing mouse Immune liver injuries to be created as
Work(.Compared with model group, leaf of Moringa granule of the invention is high, neutralize ALT and AST enzymatic activitys in the mice serum of low dose group
It is decreased obviously (P<0.05, P<0.01) it is, the most notable with the decline of high dose group, in positive control drug DDB group mice serum
ALT and AST enzymatic activitys are also than the low (P of ALT and AST enzymatic activitys in model group mice serum<0.01)；From each group mice serum ALT
Seen with AST activity, leaf of Moringa granule can reduce the extent of damage of Immune liver injury mouse liver cell, and three agent
Amount group is in certain dosage correlation.Therefore, leaf of Moringa granule of the invention is shown to immunological liver injury in mice model
Obvious hepatoprotective effect.
The leaf of Moringa granule of table 18 influences on immunological liver injury in mice hepatic tissue MDA, SOD and GSH-px
Note：Compared with model group, * represents P<0.05, * * represents P<0.01.Compared with blank group, # represents P<0.05, ##
Represent P<0.01.
Drawn, compared with blank control group mouse by table 18, MDA (MDA) content is obvious in model group murine liver tissue
Rise, with significant significant difference (P<0.05), and SOD enzymes and GSH-px enzymatic activitys are significantly lower than blank control group (P<
0.01).After leaf of Moringa granule high, medium and low dosage processing Immune liver injury mouse, from hepatic tissue SOD enzymes and
GSH-px enzymatic activitys see that the high, medium and low dosage group Mouse Liver SOD enzyme activity of leaf of Moringa granule and GSH-px enzymatic activitys are obvious
Rise (P<0.05, P<0.01), wherein leaf of Moringa granule is high, middle dose group Mouse Liver SOD enzyme activity and GSH-px enzyme activity
(128.33±14.88(U/mgpro)、120.19±16.09(U/mgpro)、30.24±3.18(mg/gpro)、31.85±
5.85 (mg/gpro)) it is close (118.04 ± 10.05 (U/mgpro), 33.51 ± 5.35 (mg/ with blank control group
gpro))。
Therefore, compared with model group, leaf of Moringa granule of the invention has humidification to liver SOD and GSH-px enzyme activity
(P<0.05, P<0.01), be conducive to improving the hepatic injury of model mice, leaf of Moringa granule has immunological liver injury protection work(
Can effect.
Above-described embodiment is preferably embodiment, but embodiments of the present invention are not by above-described embodiment of the invention
Limitation, other any Spirit Essences without departing from the present invention and the change made under principle, modification, replacement, is combined and simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of leaf of Moringa granule, it is characterised in that comprise the following specific steps that：
S1. get the raw materials ready：Leaf of Moringa is dried, crush and sieved, leaf of Moringa powder is obtained；
S2. extract：Add water in leaf of Moringa powder, extracted at 60~100 DEG C, obtain the Aqueous extracts of leaf of Moringa；
S3. purify：The Aqueous extracts of leaf of Moringa are added in ethanol and precipitated, supernatant is taken after centrifugation, as leaf of Moringa is pure
Change liquid；
S4. concentrate：Leaf of Moringa refined solution is concentrated under reduced pressure under the conditions of 60~80 DEG C, leaf of Moringa thick paste is obtained；
S5. dry：Leaf of Moringa thick paste is dried in vacuo at 60~80 DEG C, leaf of Moringa dry extract is obtained；
S6. pelletize：By leaf of Moringa dry extract and auxiliary materials and mixing, leaf of Moringa granule is obtained using wet granulation.
2. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that the mesh sieved described in step S1
Number is 80~100 mesh.
3. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that leaf of Moringa described in step S2
The mass ratio of powder and water is 1：(15~25)；The time of the extraction is 0.5~1.5h；The number of times of the extraction is 1~3 time.
4. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that leaf of Moringa described in step S3
Aqueous extracts and ethanol volume ratio be 1:(1~3)；The concentration of the ethanol is 60%~80%；The rotating speed of the centrifugation is
8000~14000rpm；The time of the centrifugation is 10~20min.
5. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that depressurized described in step S4
Vacuum is -0.06~0.07MPa；Vacuum drying vacuum described in step S5 is -0.06~0.07MPa；The Moringa
The relative density of leaf thick paste is 1.30~1.35.
6. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that auxiliary material is described in step S6
Icing Sugar and mannitol.
7. the preparation method of leaf of Moringa granule according to claim 6, it is characterised in that the Icing Sugar and mannitol
Mass ratio is (1~5):(5~1).
8. the preparation method of leaf of Moringa granule according to claim 1, it is characterised in that leaf of Moringa described in step S6
The mass ratio of dry extract and auxiliary material is (1~3):(3~5).
9. the leaf of Moringa granule prepared according to any one of claim 1-8 methods described.
10. leaf of Moringa granule described in claim 9 is in the application in preventing and treating immunological liver injury.
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| CN109123672A (en) * | 2018-08-28 | 2019-01-04 | 华南农业大学 | A kind of Moringa composition, granule, pressed candy and the preparation method and application thereof |
| CN110596263A (en) * | 2019-08-23 | 2019-12-20 | 广州泽力医药科技有限公司 | Establishing method of moringa oleifera extract fingerprint and fingerprint thereof |
| CN114304630A (en) * | 2021-12-24 | 2022-04-12 | 中国热带农业科学院热带生物技术研究所 | Rapid preparation method and application of moringa oleifera total flavonoids |
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| CN106474145A (en) * | 2016-11-29 | 2017-03-08 | 华南农业大学 | Application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109123672A (en) * | 2018-08-28 | 2019-01-04 | 华南农业大学 | A kind of Moringa composition, granule, pressed candy and the preparation method and application thereof |
| CN110596263A (en) * | 2019-08-23 | 2019-12-20 | 广州泽力医药科技有限公司 | Establishing method of moringa oleifera extract fingerprint and fingerprint thereof |
| CN114304630A (en) * | 2021-12-24 | 2022-04-12 | 中国热带农业科学院热带生物技术研究所 | Rapid preparation method and application of moringa oleifera total flavonoids |
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Application publication date: 20171010 |