CN1209155C - Balsam pear products containing multiple hypoglycemic active composition and preparation thereof - Google Patents

Balsam pear products containing multiple hypoglycemic active composition and preparation thereof Download PDF

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CN1209155C
CN1209155C CN 03128089 CN03128089A CN1209155C CN 1209155 C CN1209155 C CN 1209155C CN 03128089 CN03128089 CN 03128089 CN 03128089 A CN03128089 A CN 03128089A CN 1209155 C CN1209155 C CN 1209155C
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momordicae charantiae
fructus momordicae
goods
preparation
filtration
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CN1456337A (en
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张厚瑞
梁小燕
阮俊
陈海珊
李典鹏
曾健智
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Guilin futor Technology Consulting Co.,Ltd.
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Guilin Futor Technology Consulting Co ltd
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Abstract

The present invention provides a bitter melon product containing a plurality of hypoglycemic effect active ingredients and a preparation method thereof. The bitter melon product is a lyophilized product of a microfiltration trapped product and a nanometer filtration trapped product after bitter melon juice is made by using bitter melons as a raw material, wherein enzyme activity of the fresh bitter melon, various kinds of protein and saponin in the fresh bitter melon are preserved. The present invention has the preparation method that seeds are removed from the fresh bitter melon, and defibrination enzyme treatment and filtration for discarding coarse fibre are carried out. Then, the bitter melon is subsided and centrifugally separated to obtain bitter melon juice which adopts microfiltration and nanometer filtration separation, and microfiltration cut-off liquid is obtained to merge with the nanometer filtration cut-off liquid. After freeze-drying, hypoglycemic effect substances are obtained, a proper quantity of supplementary materials is added, and the hypoglycemic effect substances are filled into capsules, or stamped into tablets, made into granules, made into buccal tablets or made into pills for diabetic to take.

Description

The preparation method that contains the Fructus Momordicae charantiae goods of blood-sugar decreasing active
Technical field
The present invention relates to a kind of Chinese medicine with blood sugar lowering, particularly is the preparation method that raw material extracts the Chinese medicine preparation of the treatment diabetes that obtain with the Fructus Momordicae charantiae.
Background technology
Diabetes are important diseases of current harm humans health, and the whole world has the people of 5%-7% to be subjected to diabetes mellitus approximately at present, is considered to after cardiovascular and cerebrovascular disease, malignant tumor, cause human wounded or disabled, dead the third-largest killer.
Diabetes are divided into I type and II type, and type i diabetes is a kind of autoimmune disease, and patient's survival needs insulin injection.Type ii diabetes accounts for about 95% of numerous diabetics sums.The reason of type ii diabetes is an islet cells metabolism generation obstacle, hypoinsulinism, perhaps producing insulin resistant, is to be main companion's insulin secretion relative deficiency with the insulin resistant, serves as the metabolic disease that main companion's insulin resistant causes with the insulin secretion relative deficiency perhaps.Hypoinsulinism perhaps produces insulin resistant and all can cause in the blood glucose excessive, and when blood sugar content surpasses the kidney presentation time, glucose is just transferred in the urine and discharged, thereby produces dextrosuria.
The several drugs of clinical practice at present all can not be cured diabetes, mainly is by different mechanism of action, reduces patient's blood sugar level.Diabetics must be taken medicine throughout one's life, and medicine itself is threatening diabetes patient's health equally to the toxic and side effects of human body.In China, use at present medicine can blood sugar control to reach arm's length standard, 20% (the Zou Dajin that is less than the diabetics sum, chief culprit-the insulin resistant of type 2 diabetes mellitus, Popular Medicine, 2002, (6): 17), directly caused the complication of many diabeticss to take place early, progress is fast.Obviously, except screening more safely and effectively the newtype drug, seeking other the effective glucose level control of auxiliary treatment means, is crucial for the health level that improves the diabetes patient equally.
Fructus Momordicae charantiae (Momordica charantia) is that a kind of mankind have the edible historical vegetable of long-term safety.Recent two decades comes, people have found many compositions with hypoglycemic activity in the fruit of Fructus Momordicae charantiae, the extract of some Fructus Momordicae charantiaes, perhaps directly with the tea beverage of making after the Fructus Momordicae charantiae drying, recommended the diabetes patient and used, some Fructus Momordicae charantiae composition has been applied the patent as the treatment diabetes medicament.But, from scientific and technical literature, introduce and existing patent or product, mainly be based on in the many blood-sugar decreasing actives of Fructus Momordicae charantiae to certain understanding of principal component, do not recognize, have in the Fructus Momordicae charantiae composition of hypoglycemic activity known just have multiple, and chemical property has nothing in common with each other, and in addition, it is unknown also having some to have hypoglycemic active component in the Fructus Momordicae charantiae.So, existing document, patent or product, its limitation also is tangible.
(CN 98805001, the Orally active composition in the Fructus Momordicae charantiae, its bioactive peptide, and the purposes of their treatment diabetes for United States Patent (USP) (us 6391854B1) and Chinese patent;<applicant〉U.S. Calyx Therapeutics Inc) provide a kind of water solubility extract of Fructus Momordicae charantiae to be called MC6, it contains three kinds of peptides in the feature of MC6, the feature of its preparation method is: the Succus Momordicae charantiae of centrifugal clarification passes through 30kDal, the ultrafilter membrane of 10kDal and 3kDal successively.The MC6 molecular weight can pass through the 10kDal ultrafilter membrane, and be the 3kDal membrane retention less than 10kDal.The feature of activated MC6 is to move with single strips on the SDS-PAGE gel electrophoresis, and molecular weight comprises three kinds of peptides less than 10kDal.The peptide components that also has MC6 that provides simultaneously is called MC6.1, and the derivant of MC6.1 and analogies.Active MC6, MC6.1, MC6.2, MC6.3 have hypoglycemic activity, diabetes especially, and wherein preferred oral gives these active medicines.In this patent, the balsam pear hypoglycemic survivor of a murder attempt who is utilized also has this activity after obeying administration.The method composition that utilizes these active medicine treatment hyperglycemia diseases in addition that provides simultaneously only is the water-solubility protein (polypeptide) that enough sees through ultrafilter membrane.Patent application person describes, and MC6 does not improve the activity of insulin level in the animal pattern blood plasma.
The invention of international patent application (PCT/IN99/00052 1999.9.28) provides a kind of efficient blood sugar lowing polypeptide-k first, and it extracts from Fructus Momordicae charantiae.Polypeptide-k preparation method that patent application person introduces is: handle Fructus Momordicae charantiae seed (or fruit) with non-polar solven, remove unwanted oils and fats, flavonoid and sapogenin, use the protein in moisture 80% the acetone extraction residue then, optionally Crystallization Separation goes out polypeptide-k at last.The applicant describes, and polypeptide-k is water insoluble, and its hypoglycemic mechanism of action is to activate the insulin of non-activity in the blood.
Clearly, MC6 and polypeptide-k are the protein that has hypoglycemic activity in the Fructus Momordicae charantiae, and still, MC6 is water-soluble, and polypeptide-k is water insoluble, according to the preparation method of MC6, has then lost the polypeptide-k in the Fructus Momordicae charantiae; And, lost the MC6 in the Fructus Momordicae charantiae equally according to the preparation method of polypeptide-k.
Some Chinese patents disclose the content of extracting sugar reducing substance or make Chinese medicine from Fructus Momordicae charantiae in addition:
1,<and application number〉99115925<denomination of invention〉a kind of bitter gourd sugar-reducing capsule and preparation technology thereof, this invention is to be raw material with fresh Fructus Momordicae charantiae, and is freezing through cleaning, low temperature slurrying, filter pressing is crossed 100 mesh sieves with the filtering residue drying, makes the Fructus Momordicae charantiae ground-slag; Filtrate clarification refilters, and stirs 3 hours under 15 ℃ of conditions through acid (pH3.0) ethanol, staticly settled 12 hours, after the filtration with precipitate through cold drying, mistake 100 mesh sieves are made bitter melon protein; Filtrate is through 60 ℃ of vacuum concentration, add 2 times of water dissolutioies, add 2 times of ether defattings, 50 ℃ of vacuum concentration reclaim ether, and 60 ℃ of vacuum reclaim ethanol and make thick Fructus Momordicae charantiae total saponins, again through anhydrous propanone washing of precipitate several under 4 ℃ of conditions, vacuum concentration below 30 ℃ is made the Fructus Momordicae charantiae total saponins, with total saponins, bitter melon protein powder and Fructus Momordicae charantiae slag, incapsulate and make bitter gourd sugar-reducing capsule through mixing by 10: 2: 88 weight ratios at last.It is extensive to have raw material sources, and technology is simple, taking convenience, and low price, the advantage that has no side effect can be as the auxiliary agent of treatment diabetes.
2,<application number 98110802<Granted publication day 08 month calendar year 2001 15 days<denomination of invention medicine of reducing blood sugar using natural bitter gourd extract and preparation method thereof, disclosure of the Invention a kind of manufacture method of medicine of reducing blood sugar using natural bitter gourd extract, comprise the steps: that a. cuts into slices ripe Fructus Momordicae charantiae, and carry out dried; B. exsiccant Fructus Momordicae charantiae slice is soaked in the water, and extracts immersion; C. the Fructus Momordicae charantiae immersion is condensed into concentrated solution; D. concentrated solution is carried out biological enzymolysis, get the high activity degradation solution; E. degradation solution is carried out the freeze-day with constant temperature granulation of dusting; F. granule is incapsulated patent medicine; This method process equipment is simple, meets the large-scale production requirement, product yield height, and therapeutic effect is good, has no side effect.
3,<application number 95103732<denomination of invention the treatment diabetes Chinese medicine and production method thereof, this invention is a kind of Chinese medicine for the treatment of diabetes, the main component of this Chinese medicine is animal pancreas, Fructus Momordicae charantiae, Fructus Cucurbitae moschatae, the Radix Astragali, Rhizoma Dioscoreae, production method is that animal pancreas is pulverized drying, mix with the Fructus Momordicae charantiae that obtains with water extraction or separating out alcohol method, Fructus Cucurbitae moschatae, Radix Astragali extract and to add Rhizoma Dioscoreae powder again and obtain, this medicine has supplementing QI for promoting the production of body fluid, spleen invigorating is moisturized, nourishing YIN to lower pathogenic fire power is imitated, and the human body immunity improving function is arranged, promote insulin secretion, the blood sugar lowering effect.
4,<and application number〉93119351.6<denomination of invention〉new component PA of plant hypoglycemic active protein and preparation thereof, this invention relates to field of biological pharmacy.From the vegetable material Fructus Momordicae charantiae, extract the new component PA of hypoglycemic active protein, material purified treatment-cryogenic freezing among the preparation technology-fragmentation-acid alcohol dipping-high-speed homogenization-acid alcohol extraction-sucking filtration-centrifugal clarification-precipitation-dissolving-Sephadex gel is crossed the preparation method of post-ultraviolet detection fraction collection-concentrated-new component PA of exsiccant balsam pear hypoglycemic activated protein again, has been characterized in adopting isoelectric precipitation and gel to cross the method for post, ultraviolet detection fraction collection.The inventor think this material have separation and Extraction easy, consume few, the yield advantages of higher of organic solvent, obvious through animal experiment proof blood sugar decreasing effect.
5,<and application number〉02102754<denomination of invention〉a kind of method of rapidly and efficiently extracting plant insuline, the Fructus Momordicae charantiae seed contains more rich plant insuline, is a kind of effective ingredient for the treatment of diabetes at present.The invention technical characterictic is: the shelling of Fructus Momordicae charantiae seed, kernel was pulverized 60 or 100 mesh sieves, adopt acidic ethanol to extract then, the centrifuging and taking supernatant, add NaCl and account for volume 3%~5%, with 1~5Mol/LNaOH regulator solution to pH value 7.0 ± 7.5 solution becomes muddinesses, centrifugal, remove liquid, precipitate part three times with washing with acetone, vacuum drying obtains the plant insuline crude product.With 0.1MTris.cl solution (pH7.5) dissolving carrying out gel affinity chromatograph, collect eluting peak solution, concentrate vacuum drying with acetone or alcohol and obtain the plant insuline elaboration.This method can be extracted the highly purified plant insuline composition of preparation, can be used for treatment of diabetes.
6,<and application number〉98102198.0<denomination of invention〉a kind of balsam pear buccal lozenge and preparation method thereof, this invention is a kind of balsam pear buccal lozenge and preparation method thereof.This balsam pear buccal lozenge mainly contains Fructus Momordicae charantiae powder, dextrin, citric acid and mannitol etc., during preparation, earlier with bright Fructus Momordicae charantiae through squeeze the juice, filtration, lyophilization make balsam pear freeze-dried powder, mix with other composition again, and compacting is in flakes.Adopt preparation method of the present invention scientific and reasonable, fully kept the multiple nutritional components in the Fructus Momordicae charantiae, do not lose the activity of Fructus Momordicae charantiae under the made buccal tablet room temperature, have effects such as heat-clearing and toxic substances removing, the chest stuffiness relieving promoting the circulation of QI, promoting the production of body fluid to quench thirst, blood sugar lowering, raising immunity, anticancer, it is daily life, particularly in disaster area rescue, military operation and tourism, exploring, can regain one's strength of body rapidly, improve the necessary that makes things convenient for of immunity.Principal claim (A61K35/78) requires: a kind of balsam pear buccal lozenge is characterized in that containing Fructus Momordicae charantiae powder, dextrin, mannitol, spice, pigment and magnesium stearate.
Existing studies show that, outside the isolating protein, saponin also is a kind of very effectively composition of hypoglycemic activity in the Fructus Momordicae charantiae.
Some scientific and technical literature reports, the Fructus Momordicae charantiae saponin is a tetracyclic triterpenoid, and a plurality of heterogeneities are arranged, all be insoluble in water, available ethanol extracts (Chang Fenggang: Fructus Momordicae charantiae chemical constitution study, Chinese herbal medicine from exsiccant Fructus Momordicae charantiae, 1995,26 (10): 507-510).The Fructus Momordicae charantiae saponin does not influence insulin content in the animal subject model blood plasma, mainly be the synthetic (Wang Xianyuan that promotes to increase muscle glycogen and hepatic glycogen, Jin Hong, Xu Zhiqin etc.: Fructus Momordicae charantiae saponin hypoglycemic activity and Initial Study of Mechanism, aminoacid and living resources, 2001,23 (3): 42-45), thereby have significant hypoglycemic activity (Sun Shuqing, the pharmacology of Fructus Momordicae charantiae and clinical research overview, a Chinese crude drug, 1997, (8): 428--429), animal experiment proves that its blood glucose reduces amplitude even is better than " glyburide " (Sun Baoying etc., the pharmacodynamic experiment research of Fructus Momordicae charantiae saponin, Henan Chinese medicine academic periodical, 1994,9 (6): 19--21).
Water-soluble polypeptide MC6, water-insoluble polypeptide-k and Fructus Momordicae charantiae saponin, because the difference of their chemical property, thereby belong to the material that differs from one another.
And the balsam pear hypoglycemic active component not merely is known saponin and protein.
From the water cooking liquid of fresh Fructus Momordicae charantiae, isolated the composition that dissolves in ether and be insoluble to two the unknowns of ether, identified and be not saponin, flavone and alkaloid, these two kinds of compositions all show obvious functions of blood sugar activity (Wang Yongqing: balsam pear hypoglycemic effect research, Hunan Chinese medicine magazine, 1998, (6): 51-52).Prove that thus blood sugar reducing component also has other hypoglycemic activity chemical compound in the Fructus Momordicae charantiae except that saponin and vegetable protein.
Several distinct methods extracts of Fructus Momordicae charantiae hypoglycemic effect to the inductive chemical diabetes model mice of alloxan is compared, the result shows, the fruit juice of the not heated direct squeezing of Fructus Momordicae charantiae is better than the blood sugar decreasing effect of the effective site that other method is extracted, and infers that its hypoglycemic activity thing may be the water miscible easy ruined composition (Fan Yuling that is heated; Cui Fude, the comparative study of Fructus Momordicae charantiae effective site hypoglycemic activity, Shenyang Pharmaceutical University's journal 2,001 18 (1) 50-52).
Zoopery shows, bright Succus Momordicae charantiae can enable alloxan induce diabetic mice the beta Cell of islet of damaged obtain repairing, and the β cell of regenerating and making new advances, improved the insulin content (Ahmed in the blood plasma, I., Adeghate, E., Sharma, A.K., Pallot, D.J.et al.Effects of Momordica charantia fruit juice on isletmorphology in the pancreas of the streptozotocin-diabetic rat.Diabetes researchand clinical practice.1998,40,145-151).What body was interior and stripped further studies show that; this mechanism of action of bright Succus Momordicae charantiae is the pancreas that has alleviated diabetic mice; the lipid peroxidation of RIN cell and islets of langerhans; and reduced the inductive apoptosis of alloxan in the RIN cell; thereby to the RIN cell; shield (Sitasawad, S.L., the Shewade.Y of islets of langerhans and pancreatic beta cell.,Bhonde,R。,Role?of?bittergourd?fruit?juice?instz-induced?diabetic?state?in?vivo?and?in?vitro.Journal?of?Ethno-pharmacology,2000,73(1):71-79)。And in the known hypoglycemic activity chemical compound of Fructus Momordicae charantiae, still fail to determine to have this reparation or the islet cells of regenerating, improve the biological activity of insulin level.
By the above-mentioned fact as can be known, the function of blood sugar reduction active matter of Fructus Momordicae charantiae has knownly, also belongs to unknown in addition, the function of blood sugar reduction of Fructus Momordicae charantiae, and actual is the wherein contained multiple synergistic result of composition with hypoglycemic activity.With regard to known active component,, be difficult to they be extracted simultaneously with a kind of solvent because of its chemical property difference.In addition, in the hypoglycemic activity thing of Fructus Momordicae charantiae, include heat labile composition, the process of its extraction should be heated.
With the way of defibrination that fresh Fructus Momordicae charantiae pulp organization cell is fully broken, all inclusions of cell comprise all being released to water-soluble and the blood-sugar decreasing active that is insoluble in water in the pulp.With the crude fibre in the pulp (coming from the flesh cell wall) elimination, lyophilization then, the freeze-dried products of this Succus Momordicae charantiae has comprised undoubtedly than more comprehensive blood-sugar decreasing active.But the content of hypoglycemic activity thing is very low in fact in the Succus Momordicae charantiae, direct lyophilizing cost costliness, and on the other hand, the glucide that exists in the Succus Momordicae charantiae is deliquescence very easily, makes this freeze-dried products packing under normal operation, storage and use at all.Though add the deliquescence problem that a large amount of drying aids or anti-blushing agent can solve the freeze-dried products of Succus Momordicae charantiae, can't make high-load balsam pear hypoglycemic goods.
In sum, the preparation of the Fructus Momordicae charantiae effective ingredient of document and patent disclosure, basically be to adopt the physics method and chemical method extracts certain or several clear and definite chemical constituents are main, or Fructus Momordicae charantiae certain solvent under heating condition is (as water, ethanol or its mixture) total extract, they or the not mentioned blood-sugar decreasing active that how to utilize those chemical property the unknowns in the Fructus Momordicae charantiae, some thermal sensitivity hypoglycemic activity thing is subjected to the heat damage problem in the perhaps not clear and definite Fructus Momordicae charantiae.
No. 99115925 Chinese invention patents, a kind of bitter gourd sugar-reducing capsule and preparation technology thereof have proposed the utilization of bitter melon protein and Fructus Momordicae charantiae saponin, but have used ethanol in preparation process, acetone, organic solvents such as ether.No. 98110802 patents are mentioned in medicine of reducing blood sugar using natural bitter gourd extract and preparation method thereof the Fructus Momordicae charantiae concentrated solution are carried out biological enzymolysis, the high activity degradation solution, but what enzyme what adopt in the not mentioned biological enzymolysis is.98102198.0 what use in number patent, a kind of balsam pear buccal lozenge and preparation method thereof is lyophilization Succus Momordicae charantiae technology, but it does not consider to remove the known problem that does not have the composition of hypoglycemic activity in the Fructus Momordicae charantiae.
For these reasons, the different all kinds of hypoglycemic activity things of physicochemical property in the exploitation one cover energy full use Fructus Momordicae charantiae, and active component content height, the preparation method of the balsam pear hypoglycemic goods that physicochemical property is stable, be not only the only way of the highly active balsam pear hypoglycemic goods of exploitation, and, also be very necessary for fully and reasonably utilizing the Fructus Momordicae charantiae resource.
Technology contents
The present invention is primarily aimed at and contains multiple blood-sugar decreasing active in the Fructus Momordicae charantiae, its chemical property has nothing in common with each other: have water-soluble, also in distress water-soluble and be soluble in polar organic solvent, the then water insoluble and polar organic solvent of some composition, and can only be dissolved in non-polar organic solvent, thereby all can not extract whole active component with any solvent (comprising water) comprehensively, thermo-labile this problem of some unknown active component proposes simultaneously.A cover balsam pear hypoglycemic active matter preparation technology who is provided has not only utilized the various blood-sugar decreasing actives in the Fructus Momordicae charantiae all sidedly, and has removed known nonactive thing to greatest extent, has avoided heat loss's problem of thermal sensitivity active matter simultaneously.Thereby adopt Fructus Momordicae charantiae goods of the present invention, and hypoglycemic activity thing content height, using dosage is little and hypoglycemic activity outstanding, and processing characteristics is stable.
Technical scheme of the present invention is as follows:
The Fructus Momordicae charantiae goods that contain a plurality of blood-sugar decreasing actives of the present invention, it comprises the enzymatic activity in the bright Fructus Momordicae charantiae that exists in the goods, multiple proteins, the main saponin constituent in the bright Fructus Momordicae charantiae in the bright Fructus Momordicae charantiae and remove known nonactive thing to greatest extent, that concrete is is fresh, through the Fructus Momordicae charantiae slurry of enzymolysis processing, after filtration, the microfiltration trapped substance and the nanofiltration trapped substance of gained Succus Momordicae charantiae after the centrifugalize.
The above-described enzymatic activity that contains the Fructus Momordicae charantiae goods of a plurality of blood-sugar decreasing actives is meant with the polyphenol oxidase enzyme activity to serve as the enzymatic activity of mark Chi.
Above-described multiple proteins is the material of protein spectrum basically identical that exist in the goods, in its protein native polyacrylamide gel electrophoresis spectrum and the bright Fructus Momordicae charantiae.
Above-described saponin is a material that exist in the goods, its thin layer chromatogram the has feature speckle consistent with bright Fructus Momordicae charantiae raw material saponin chromatograph.
Above-described known nonactive thing glucose content (to the sample soluble solid) is lower than the glucose sugar content (to bright Succus Momordicae charantiae soluble solid) of bright Succus Momordicae charantiae.
Above-described fresh Fructus Momordicae charantiae slurry is that fresh Fructus Momordicae charantiae is gone to beat the juice that (mill) made with beating (mill) pulp grinder behind the seed; Enzymolysis processing is meant in Fructus Momordicae charantiae slurry and adds cellulase, hemicellulase, a kind of in the pectase, or their mixture, the pectin in cellulose, hemicellulose and the pulp of enzymolysis Fructus Momordicae charantiae cell wall.
Above-described microfiltration trapped substance is that the Fructus Momordicae charantiae after the enzymolysis processing is starched after filtration or the centrifugal sarcocarp residue of removing, and what the gained Succus Momordicae charantiae adopted the micro-filtration collection can not be the canescence insoluble matter of 0.2 μ m-0.45 μ m microporous filter membrane through the aperture.
Above-described nanofiltration trapped substance is meant and can sees through 0.2 μ m-0.45 μ m microporous filter membrane, is the material that the NF membrane of 0.2--1Kdal is held back but can be molecular cut off.
The preparation method that contains the Fructus Momordicae charantiae goods of a plurality of blood-sugar decreasing actives of the present invention comprises following step:
1, at first fresh Fructus Momordicae charantiae is removed seed, existing mill (beating) pulp grinder tool is fully broken with the Fructus Momordicae charantiae meat tissue in the employing food industry, and Fructus Momordicae charantiae flesh cell inclusions can fully be separated out, and must have the Fructus Momordicae charantiae juice of ground flesh.
2, in above-mentioned Fructus Momordicae charantiae juice, add about 0.02%-0.08% pectase, cellulase or hemicellulase, the 1-4 hour time of insulation under 30-40 ℃ of condition, remove the pulp residue then after filtration, sedimentation and centrifugation is removed the pulp particle in small, broken bits that remains in the fruit juice, draws a kind of muddy Succus Momordicae charantiae that contains pale powder shape float.Through check, the main component of this canescence float is the Fructus Momordicae charantiae saponin.
3, adopt micro-filtration (pore size filter 0.2 μ m-0.45 μ m) to filter the above-mentioned Succus Momordicae charantiae that contains the canescence float, collection can not see through the canescence insoluble matter of filter, gets component A.
4, the above-mentioned clarification Succus Momordicae charantiae that sees through micro-filtration further separates with the nanofiltration separation device, and reservation can not see through the solute of molecular cut off 0.2--1Kdal NF membrane, removes and deenergizes through saccharide, the organic acid of filter membrane.The nanofiltration later stage adds pure water repeatedly in trapped fluid, continue the nanofiltration operation, to remove sugar, the organic acid in the trapped fluid comparatively fully.Collect the nanofiltration trapped fluid, get B component.
5, the B component with the component A of above-mentioned steps 3, gained and step 4, gained merges the lyophilization collection.Obtain containing the goods of a plurality of balsam pear hypoglycemic active component.
This goods that contain a plurality of balsam pear hypoglycemic active component of the present invention have the obvious functions of blood sugar activity, can be direct, or add appropriate amount of auxiliary materials as required, fill becomes capsule, or strikes out tablet, makes granule, makes buccal tablet, makes pill, for diabetic to take.The dry thing of the sarcocarp in small, broken bits that described adjuvant is above-mentioned preparation method step 2, centrifugation obtained and food or the excipient substance that adopts existing national standard, the international standards organization standard generalized markup.
Goods of the present invention have comprised bright Fructus Momordicae charantiae to be removed after the pulp residue, the whole water insoluble active ingredients except that chlorophyll in the fruit juice, and remove the water soluble ingredient behind saccharide, the organic acid and by force can not see through the micromolecule composition of NF membrane because of hydrophobicity.Because goods of the present invention are complete operations under room temperature or freezing conditions, thereby preserve some the enzymatic activity feature in the fresh fruit.
The chemical characteristic of goods ingredient of the present invention, the enzymatic activity that is kept, can detect confirmation respectively by following method:
1. polyphenol oxidase activity detects
Goods 1g of the present invention is dissolved in the 10ml distilled water, and is centrifugal, gets supernatant 1ml, adds 0.3% pyrogallic acid aqueous solution 3ml, drips 4 low concentration hydrogen peroxide, and brown reaction takes place promptly visible solution, confirms to have polyphenol oxidase activity in the goods.
2. protein electrophorese detects
Goods 1g of the present invention is dissolved in distilled water 10ml, and is centrifugal, and get supernatant and carry out native polyacrylamide gel electrophoresis, and with Fructus Momordicae charantiae sarcocarp natural juice in contrast.Gel behind the electrophoresis is through coomassie brilliant blue staining, and the protein spectrum of goods of the present invention and Fructus Momordicae charantiae sarcocarp natural juice protein spectrum basically identical are seen Fig. 2..Show that goods of the present invention have comprised the multiple proteins in the bright Fructus Momordicae charantiae.
3. Fructus Momordicae charantiae saponin thin layer chromatography detects
Goods 1g of the present invention is dissolved in methanol 2ml, gets the methanol solution point sample on the efficient thin plate of silica gel G, does developing solvent after closing so that chloroform-methanol (7: 2.5) water is full, launches, and takes out, and dries the colour developing of 10% sulphuric acid liquid spraying post-heating.Other gets the methanolic extract point sample of Fructus Momordicae charantiae raw material on same thin plate.Goods of the present invention have 1,2 two feature speckle consistent with Fructus Momordicae charantiae raw material chromatograph, see Fig. 3..Confirm that goods of the present invention have kept saponin constituent main in the Fructus Momordicae charantiae.
4. the saccharide chromatograph detects
Goods of the present invention are dissolved in distilled water, and centrifugal disgorging, adding distil water are adjusted to soluble solid content 2.5% (refractometer mensuration).Other squeezes bright Succus Momordicae charantiae, also with distilled water soluble solid content is adjusted to 2.5%.Through Benson carbohydrate post (BC-100, Ca + 2) carry out high performance liquid chromatography (HPLC) column chromatography, goods of the present invention and the equal eluting of bright Succus Momordicae charantiae are two peaks, and first peak flowed out in the time of 9.7 minutes, and second peak (glucose) flowed out in the time of 15.0 minutes, sugar chromatographic peak retention time overlaps fully, as can be known both contained saccharide unanimities.With identical soluble solid content is benchmark, characterizes relative sugar content with the glucose absorption peak area, and the relative sugar content of goods of the present invention is lower than bright Succus Momordicae charantiae, confirms that goods of the present invention have passed through the desaccharide PROCESS FOR TREATMENT.
Description of drawings
Fig. 1. be process chart of the present invention.
Fig. 2. be the protein polyacrylate hydrogel electrophoresis detection figure as a result of goods of the present invention.
Fig. 3. be the Fructus Momordicae charantiae saponin thin layer chromatography testing result figure of goods of the present invention.
Fig. 4 .-Fig. 9. be the blood sugar decreasing effect figure of goods of the present invention.
Recognize from Fig. 1: the production technology of goods of the present invention is that fresh balsam pear is removed seed, and defibrination, pulp add enzyme preparation and protect Temperature 1-4 hour is filtered and is discarded the pulp residue, and the Bitter Melon Juice of collection (1) is again through the centrifugal removal sediment of decanter type, the balsam pear that obtains Juice (2). Bitter Melon Juice (2) gets the Bitter Melon Juice (3) that micro-filtration dams liquid (A) and clarifies through micro-filtration, and Bitter Melon Juice (3) is used nanofiltration device Separate, discard nanofiltration and see through liquid, reclaim dam liquid (B) and merge with the micro-filtration liquid (A) that dams of nanofiltration, obtain hypoglycemic system after the freeze-drying The product thing. This hypoglycemic goods can be directly or are added pulp in small, broken bits that above-mentioned centrifugation obtains, also can add other symbol Close the auxiliary material of country and international standard and make all kinds of Chinese medicine preparations, become capsule such as can, strike out tablet, or the granulation agent, For diabetic to take.
Fig. 2. shown the native polyacrylamide gel electrophoresis protein spectrum of goods of the present invention and fresh Bitter Melon Juice, both eggs White matter colour band position is basically identical, detects the multiple proteins that has comprised in the proved invention goods in the bright balsam pear with this.
Fig. 3. shown the consistent feature spot in thin-layer chromatography of balsam pear saponin(e in goods of the present invention and the balsam pear raw material, examined with this Survey proved invention goods and kept saponin constituent main in the balsam pear.
Fig. 4. provide the patient and taken the front blood sugar decline situation all around of goods of the present invention, taken goods of the present invention after one week, blood sugar Obvious decline is namely arranged, until 4th week; In eight middle of the month subsequently, the patient subtracts and has taken Western medicine, continues to take goods of the present invention, It is steadily normal that blood sugar level keeps, such as Fig. 5..
Fig. 6. illustrate the patient and take goods of the present invention front and back change of blood sugar all around, the patient withdraws all hypoglycemic medicines simultaneously, extremely After one week, blood sugar descends too much, by reduce dose half, blood sugar slightly has rising immediately, continues to take, blood sugar begins again lower Fall, effect is remarkable.
Fig. 7. illustrate the patient and take goods of the present invention front and back change of blood sugar all around, cooperate Western medicine to take, blood sugar is namely bright after the week Aobvious decline, all around after, blood sugar has been down to reduced levels.
Fig. 8. illustrate the patient and take goods of the present invention front and back change of blood sugar all around, cooperate Western medicine to take, blood sugar is namely bright after the week Aobvious decline, from the 3rd week beginning to reduce the Western medicine dose, blood sugar is still kept lower normal level.
Fig. 9. illustrate the patient and take goods of the present invention front and back change of blood sugar all around, cooperate Western medicine to take goods dose of the present invention Greater than former routine patients, blood sugar descends obviously.
The specific embodiment:
Embodiment 1
Fresh Fructus Momordicae charantiae 100Kg cuts open and removes seed, and ground flesh adds pectase 14g, 35 ℃ of insulation 2h, the centrifugal fiber residue of removing of filtering type gets light green color Succus Momordicae charantiae 70Kg, decanter type is centrifugal removes sedimentary chlorophyll, sarcocarp in small, broken bits, outward appearance be light yellow to linen muddy Succus Momordicae charantiae.
The micro-filtration that with the aperture is 0.45 μ m is carried out fine straining to muddy Succus Momordicae charantiae, during to the only remaining small size of microfiltration trapped fluid, adds an amount of pure water displacement.The muddy trapped fluid of collection behind the displacement microfiltration.
The clarification Succus Momordicae charantiae that sees through micro-filtration joins and carries out the nanofiltration processing in the nanofiltration device, and reservation can not see through the solute of molecular cut off 0.2-1Kdal NF membrane, collects the nanofiltration trapped fluid, discards the nanofiltration permeate.When the nanofiltration trapped fluid is left small size, add an amount of pure water displacement nanofiltration.
Microfiltration trapped fluid and nanofiltration trapped fluid merge lyophilization, get freeze-dried products 520g, and fill becomes capsule after the adding appropriate amount of starch.
Embodiment 2
Fresh Fructus Momordicae charantiae 150Kg, cut open and remove seed, wear into pulp, add cellulase (containing hemicellulase activity) 50 grams, pectase 30g, 40 ℃ of insulation 1h, the centrifugal fiber residue of removing of filtering type, light green color Succus Momordicae charantiae 120Kg, centrifugal that precipitate and outward appearance are light yellow to linen muddy Succus Momordicae charantiae through decanter type.
The micro-filtration that with the aperture is 0.45 μ m is carried out fine straining to muddy Succus Momordicae charantiae, when the only remaining small size of microfiltration trapped fluid, adds an amount of pure water displacement.The muddy trapped fluid of collection behind the displacement microfiltration.
The clarification Succus Momordicae charantiae that micro-filtration sees through joins nanofiltration device, and reservation can not see through the solute of molecular cut off 0.2-1Kdal NF membrane, when the nanofiltration trapped fluid is left small size, adds an amount of pure water displacement nanofiltration.Collect the nanofiltration trapped fluid, discard the nanofiltration permeate.
The sedimentation and centrifugation precipitate mixes with microfiltration trapped fluid, nanofiltration trapped fluid and lyophilization then through sterilization treatment, gets goods 1300 grams.Strike out tablet.
Application implementation 1.
Patient Mr. Zhang, 65 years old, suffer from surplus the chronic diabetes ten year, take multiple its fasting glucose of blood sugar lowering Western medicine and still can be in any more, and complication such as numbness of hands and feet, skin Pruritus, retinal hemorrhage have taken place.After adding goods of the present invention, blood glucose is reduced to normal level very soon.After one month, subtracting the clothes Western medicine, is original 1/3rd until the Western medicine dose, and the patient took goods of the present invention over eight months, and it is normal that blood sugar level keeps, and syndrome disappears, and is healthy.The change of blood sugar of taking before and after the goods of the present invention is seen Fig. 4., Fig. 5..
Application implementation 2.
Patient Liang Nvshi, 60 years old, chronic diabetes, take goods of the present invention certainly after, withdraw all hypoglycemic medicines, blood sugar level keeps normal as a result.Take the change of blood sugar of goods of the present invention front and back and see Fig. 6..
Application implementation 3.
Patient Wei Nvshi, 72 years old, diabetic history 4 years, fasting blood glucose level is 8.5mmol/L.The normal phase is taken Western medicine, take goods of the present invention after, blood sugar level steadily descends, through reducing Western medicine and regulating goods dose of the present invention, it is normal that its blood sugar level keeps.Take the change of blood sugar of goods of the present invention front and back and see Fig. 7..
Application implementation 4.
Patient Liang Nvshi, 66 years old, suffered from chronic diabetes 5 years, concurrent suffer from hypertension, blurred vision.After adding goods of the present invention, blood sugar level steadily descends, and behind the minimizing Western medicine, blood sugar level still keeps normally, and makes blood pressure controlled, and is healthy.Change of blood sugar is seen Fig. 8 before and after taking goods of the present invention.
Application implementation 5.
Patient Ms Zhang, 60 years old, suffered from diabetes 1 year, fasting blood glucose level reaches 18mmol/L, takes multiple hypoglycemic medicine poor effect, and patient's vision is fuzzy, and it is unable to have a pain in the leg, numbness of hands and feet.After taking goods of the present invention, blood glucose obviously descends, and patient feels spirit.Change of blood sugar is seen Fig. 9 before and after taking goods of the present invention..

Claims (2)

1, a kind of preparation method that contains the Fructus Momordicae charantiae goods of blood-sugar decreasing active, comprise with bright Fructus Momordicae charantiae squeeze the juice, filtration, exsiccant process, it is characterized in that: preparation method comprises following step:
(1) fresh Fructus Momordicae charantiae is removed seed, and Fructus Momordicae charantiae meat is ground pulping, and Fructus Momordicae charantiae meat cell inclusion can fully be separated out, and must have the Fructus Momordicae charantiae juice of ground flesh;
(2) in above-mentioned Fructus Momordicae charantiae juice, add 0.02%-0.08% pectase, cellulase or hemicellulase, insulation is 1-4 hour under 30-40 ℃ of condition, remove the pulp residue then after filtration, sedimentation and centrifugation is removed the pulp particle in small, broken bits that remains in the fruit juice, draws a kind of muddy Succus Momordicae charantiae that contains pale powder shape float;
(3) adopt the micro-filtration of pore size filter 0.2 μ m-0.45 μ m to filter the above-mentioned Succus Momordicae charantiae that contains the canescence float, collection can not see through the canescence insoluble matter of filter, gets component A;
(4) with the above-mentioned clarification Succus Momordicae charantiae that sees through micro-filtration, put into the nanofiltration separation device, reservation can not see through the solute of 0.2-1Kdal filter membrane, and saccharide is removed in nanofiltration, organic acid in the nanofiltration later stage, adds pure water repeatedly in trapped fluid, continue the nanofiltration operation, collect trapped fluid in the nanofiltration device, get B component;
(5) the component A of above-mentioned (3) step gained and the B component of (4) step gained are merged, lyophilization obtains containing the goods of balsam pear hypoglycemic active component.
2, the preparation method that contains the Fructus Momordicae charantiae goods of blood-sugar decreasing active according to claim 1, it is characterized in that: the goods that will contain the balsam pear hypoglycemic active component directly or after adding appropriate amount of auxiliary materials fill become capsule, or strike out tablet, make granule, make buccal tablet, make pill, described adjuvant comprises the dry thing of the sarcocarp in small, broken bits that centrifugation obtains, or meet country, international standard, physiologically acceptable excipient substance.
CN 03128089 2003-05-30 2003-05-30 Balsam pear products containing multiple hypoglycemic active composition and preparation thereof Expired - Fee Related CN1209155C (en)

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Publication number Priority date Publication date Assignee Title
CN1330265C (en) * 2004-10-28 2007-08-08 山东师范大学 Bitter gourd juice drink production method
CN101461514B (en) * 2009-01-22 2011-11-30 韩金光 Bitter melon extract preparation method
CN101485429B (en) * 2009-02-25 2013-03-27 武汉科技大学 Preparation method of hypoglycemic bitter melon freeze-dried powder
CN102835633A (en) * 2012-09-19 2012-12-26 民勤县潭龙食品有限公司 Pumpkin powder production process
CN107468744A (en) * 2017-09-15 2017-12-15 李玉保 A kind of new method, bitter gourd polypeptide protein extract and its application of whole temperature production bitter gourd polypeptide protein extract
CN109833337B (en) * 2019-01-18 2021-08-03 华南农业大学 Momordica charantia essence compound for inhibiting pest immune response and high-efficiency preparation method and application thereof
CN111171112A (en) * 2020-03-17 2020-05-19 广西壮族自治区农业科学院 Enzymatic extraction method of bitter melon seed protein

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