CN108295240A - A kind of ginseng oral liquid of invigorating qi for tranquilization and preparation method thereof - Google Patents
A kind of ginseng oral liquid of invigorating qi for tranquilization and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to field of health care products more particularly to a kind of ginseng oral liquid of invigorating qi for tranquilization and preparation method thereof.Oral solution of the present invention is prepared by ginseng total glycosides extractive, Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, Semen Platycladi extract, Fructus lycii P.E, Wild jujube seeds extract and small molecule ginseng polypeptide's chelates of zinc, have effects that repose tonifying Qi, tranquilizing the mind prevent prolapse, reinforce the spleen to benefit the lung, yin-nourishing of enriching blood, calming heart brain tonic, be suitable for the diseases such as apathetic, deficiency of both qi and yin caused by the blood deficiency of Tianjin, with a haggard face, insomnia forgetfulness, neurasthenia.Extracting technique of Chinese medicine of the present invention is efficient, at low cost, energy saving, environmentally protective, further improve the economic value of Chinese medicine, it is made oral solution high effect nontoxic, convenient to take, especially it is added to the small molecule ginseng polypeptide's chelates of zinc being easily absorbed by the body, so that user is supplemented the amino acid and trace element of needed by human body simultaneously, enriches the nutritive value of oral solution health products.
Description
Technical field
The invention belongs to field of health care products more particularly to a kind of ginseng oral liquid of invigorating qi for tranquilization and preparation method thereof.
Background technology
With the continuous quickening of rhythm of life, work and life stress continue to increase, and not only daytime will be by many working clans
The work of unit anxiety it is also often desirable to overtime work and stay up late, long-term lack of sleep, in addition diet is irregular, nutriment intake
Deficiency causes crowd's quantity of inferior health more and more.Most common problem is exactly to have a sleepless night in sub-health population, chronic insomnia meeting
There is dysphoria, attention cannot be concentrated, poor memory, often feel like the symptoms such as tired.Chinese medicine thinks that insomnia belongs to Ying Wei
It becomes estranged, the disease of imbalance of yin and yang or the deficiency of Yin cannot receive sun or the overabundance of yang must not enter the moon.It is exactly YIN and YANG balance regulating to take Chinese medicine treatment insomnia,
Its growth and decline is set to balance.
Ginseng (Panax ginseng C.A.Meyer) is Araliaceae, is traditional rare traditional Chinese medicine, is swung because it has
Long medicinal history and special efficacy is favored in depth between east and west by people since ancient times.Ginseng exists《Sheng Nong's herbal classic》Middle quilt
It is classified as top grade, Chinese medicine effect is to reinforce vital energy, invigorate the spleen and benefit qi, veins takes off admittedly, beneficial lung, promote the production of body fluid, calm the nerves, and a large amount of pharmacological researches face
Bed observation and epidemiological investigation, which confirm that ginseng has, to be improved microcirculation, improves tissue anti-anoxia ability, inhibits blood small
The multiple biological activities such as plate aggregation, antitumor, anti-aging, radioresistance, antifatigue.On September 4th, 2012, ministry of Health of China approval
Ginseng becomes new resource food, and the application of ginseng will be extended to the fields such as food, beverage and health product by single Chinese medicine.
Common ginseng health product currently on the market, or addition ginseng crude extract are unfavorable for absorption of human body active ingredient, or
Lead to that valuable raw material are largely wasted or the single health-care effect of ingredient is limited because effective component extraction rate is low.
Invention content
To solve above-mentioned the deficiencies in the prior art, the present invention provides a kind of ginseng oral liquid of invigorating qi for tranquilization and its preparations
Method.
Technical scheme of the present invention：
A kind of ginseng oral liquid of invigorating qi for tranquilization, includes the component of following parts by weight：5~10 parts of ginseng total glycosides extractive,
10~30 parts of Astragalus Root P.E, 10~30 parts of siberian Ginseng P.E, 5~20 parts of tuckahoe extracts, Semen Platycladi extract 5~20
Part, 5~20 parts of Fructus lycii P.E, 5~20 parts of Wild jujube seeds extract, small molecule ginseng polypeptide -30~80 parts of chelates of zinc；It goes
500 parts of ionized water；The preparation method step of its small molecular ginseng polypeptide-chelates of zinc enters as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, it is that neutral Tris-HCl is buffered that pH value, which is added, by certain mass volume ratio
Solution, after stirring evenly under certain temperature stirring and leaching, collect leaching liquor；It repeats to extract primary and will close leaching liquor twice
And the ultrafiltration membrane that molecular cut off is 5~30KDa is placed it in by after leaching liquor filtering clarification with 0.22~0.45 μm of filter membrane
Device carries out ultrafiltration, and it is the ginseng water-soluble protein crude extract for removing small molecular weight impurity to collect ultrafiltration to retain liquid；
S2, enzymolysis ginseng water-soluble protein prepare small molecule ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5, alkali protease is added, carries out at a certain temperature by ginseng water-soluble protein concentrate
It digests, NaOH solution maintenance system pH value is added dropwise in enzymolysis process, HCl is added by system pH in enzymolysis into enzymatic hydrolysis system after completing
5~7 are adjusted to, neutral proteinase is added, enzymatic hydrolysis system temperature is risen to 90~100 DEG C after completing enzymolysis at a certain temperature and is protected
Holding 10 minutes makes enzyme inactivate, and it is neutrality that pH value is adjusted after cooled to room temperature, obtains enzymolysis liquid；It will using SephadexG-25
The enzymolysis protein classification obtained in enzymolysis liquid, collects the small molecule ginseng polypeptide that molecular weight is 500~1000Da；
S3, small molecule ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected is uniformly mixed with zinc sulfate by certain mass ratio, by reactant
System is adjusted to certain pH value, and the chelating of small molecule ginseng polypeptide and zinc sulfate, water-bath certain time are carried out under certain bath temperature
The ethanol solution that mass concentration is 75% is added in backward system, stands 20~30min, 4000r/min centrifuges 15min, collects
Precipitation, vacuum drying obtain small molecule ginseng polypeptide-chelates of zinc.
Further, the ginseng oral liquid of the invigorating qi for tranquilization includes the component of following parts by weight：Ginseng total glycosides extractive 8
Part, the extraction of 20 parts of Astragalus Root P.E, 20 parts of siberian Ginseng P.E, 15 parts of tuckahoe extracts, 15 parts of Semen Platycladi extract, the fruit of Chinese wolfberry
15 parts of object, 15 parts of Wild jujube seeds extract, small molecule ginseng polypeptide -60 parts of chelates of zinc；500 parts of deionized water.
Further, the total glycosides purity of ginseng is 90~96% in ginseng total glycosides extractive, Astragalus Root P.E, wilsonii extraction
Object, tuckahoe extracts, Semen Platycladi extract, Fructus lycii P.E and Wild jujube seeds extract mass concentration be 80~85%.
Further, the mass volume ratio of ginseng raw material described in step S1 and Tris-HCl buffer solutions is 1g:10~
15mL；The molar concentration of the Tris-HCl buffer solutions is 0.01mol/L；The temperature of the stirring and leaching is 4 DEG C, stirring leaching
It is 3~6h to carry the time.
Further, the enzyme concentration of alkali protease described in step S2 is the 0.2~0.6% of substrate quality, hydrolysis temperature
It it is 55~70 DEG C, enzymolysis time is 1~2h；The enzyme concentration of the neutral proteinase is the 2.0~3.0% of substrate quality, enzymolysis
Temperature is 50~60 DEG C, and enzymolysis time is 1~2h.
Further, the mass ratio of small molecule ginseng polypeptide and zinc sulfate described in step S3 are 5~7:1；The pH value is
6.5~7.5, bath temperature is 50~60 DEG C, and water bath time is 30~40min.
Steps are as follows for the preparation method of the ginseng oral liquid of invigorating qi for tranquilization of the present invention：
Step 1: preparing ginseng total glycosides extractive：
Fresh ginseng flower bud is chosen, dry 10~15h in 60~65 DEG C of vacuum drying chambers is placed it in after cleaning up,
Ginseng flower bud after abundant drying is crushed to 50~100 mesh；It is 1g by mass volume ratio：15~20mL is to gained ginseng flower bud
It is 40% ethanol solution that mass concentration is added in powder, impregnates 2~3h under room temperature, will be molten immersed with the ethyl alcohol of ginseng flower bud powder
Liquid is fitted into the polybag of pressure resistance, is put into after sealing in the extraction vessel of high pressure extraction device；It will in 10~15min under room temperature
Pressure in high pressure extraction device is increased to 300~600Mpa, fills high pressure extraction in 10~20s after 15~25min of pressurize
Pressure in setting is let out rapidly as normal pressure；It repeats after high pressure extraction operates 3~5 times, by polybag from the extraction of high pressure extraction device
It is taken out in container, material filtering is removed into filter residue, filtrate is placed in 10~30min of centrifugation under 3000~5000rmp centrifugal rotational speeds,
Take supernatant repeated centrifugation 1 time, supernatant is the total glycosides crude extract of ginseng；The total glycosides crude extract of ginseng is concentrated, will be concentrated
Liquid pH value, which is adjusted to make it through macroporous absorbent resin D101 chromatographic columns after neutrality, to be adsorbed；The ethyl alcohol for being 20% by mass concentration
Solution ph is adjusted to neutrality and is eluted as the 1st eluting solvent；The ethanol solution pH value tune for being 60% by mass concentration
It is eluted to neutrality and as the 2nd eluting solvent, collects the fraction containing the total glycosides ingredient of ginseng, be concentrated under reduced pressure, spraying is dry
It is dry, obtain ginseng total glycosides extractive after purification；
Step 2: using supercritical CO2Extracting method prepares Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, cypress
Sub- benevolence extract, Fructus lycii P.E and Wild jujube seeds extract；
Step 3: respectively by following parts by weight weigh 5~10 parts of ginseng total glycosides extractive, 10~30 parts of Astragalus Root P.E,
10~30 parts of siberian Ginseng P.E, 5~20 parts of tuckahoe extracts, 5~20 parts of Semen Platycladi extract, Fructus lycii P.E 5~20
500 parts of water are added in part, 5~20 parts of Wild jujube seeds extract and small molecule ginseng polypeptide -30~80 parts of chelates of zinc, agitated mixed
Filtering and impurity removing, filling, high-temperature sterilization after closing uniformly, obtain the ginseng oral liquid of the invigorating qi for tranquilization.
Further, supercritical CO described in step 22Extracting method is：Fully dry Chinese medicine material is crushed, is taken
Chinese medicine material after 100g crushing is put into 1L extraction kettles and is extracted, supercritical CO2Fluid extraction operating process is liquid CO2→
High-pressure pump → extraction kettle (entering supercriticality) → I → separating still of separating still II → cycle；CO in extraction process2Fluid flow
For 20L/h, extraction kettle temperature is 45 DEG C, extracting pressure 30MPa, separating still I, separating still II pressure be 6MPa, separation
Temperature is room temperature, uses absolute ethyl alcohol as entrainer in extraction process, and absolute ethyl alcohol dosage is 500mL, and 2~3h of extraction is
Obtain Chinese medical extract.
Beneficial effects of the present invention：
1, the ginseng oral liquid of invigorating qi for tranquilization of the present invention is carried with the active ingredient of the rare traditional Chinese medicines such as ginseng total glycosides extractive
It is to make raw material to take object, by the mutual compatibility between each effective ingredient, oral solution is made to have tonifying Qi of reposing, calm the nerves admittedly
Take off, reinforce the spleen to benefit the lung, yin-nourishing of enriching blood, sperm adding and marrow healthing, calming heart brain tonic the effect of, be suitable for Tianjin blood deficiency caused by apathetic, gas
The diseases such as Yin bivacuity, with a haggard face, insomnia forgetfulness, neurasthenia.
The present invention goes out the active drug ingredient in Chinese medicine using advanced extractive technique high efficiency extraction, keeps the present invention oral
Liquid effective active composition content is high, and impurity is few, avoids the potential sensitization danger of non-active ingredients in traditional Chinese patent medicine drink, with
And the burden brought to user's liver kidney of complicated drug ingedient and damage, it is obtained oral solution high effect nontoxic, convenient to take.
It also added small molecule ginseng polypeptide-chelates of zinc in oral solution of the present invention, small molecule ginseng polypeptide's molecular weight exists
1000D mutually chelates small molecule ginseng polypeptide with trace element zinc hereinafter, be highly susceptible to being absorbed by the body, metal ion with
After peptide molecule is combined by coordinate bond, stable chelate is generated, not only stability is good, alleviates the antagonism between minerals
It acts on, and reduces the influence of the substances such as pH value, lipid, fiber and hydrochloric acid in gastric juice in digestion process, keep user small in absorption
Molecule ginseng polypeptide can more fully absorb while supplementing the essential amino acid that it is rich in by body, using small
The trace element chelated on molecule ginseng polypeptide.
The present invention is that Chinese medicine deep processing provides new thinking with utilization, is realizing the same of the healthcare function of invigorating qi for tranquilization
When, the effect of supplement amino acid and trace element is also acted, the nutritive value of oral solution health products is enriched.
2, high pressure extraction method used by the present invention extraction total glycosides of ginseng is sent out using osmotic pressure moment inside and outside plant cell
The structure change that cell membrane generates when changing so that the total glycosides of intracellular ginseng can be transferred to extracellularly across cell membrane
Extracting solution in, improve the rate of transform of the total glycosides of ginseng, make ginseng total glycosides extraction more fully, improve the utilization rate of ginseng,
Reduce cost of material.Compared with the extracting methods such as hot reflux, decoction, microwave, ultrasound, the present invention extracts at room temperature,
Reduce the destruction to active constituents of medicine, obtained extract have better medical active, also have extraction time it is short, effect
The advantages that rate is high, easy to operate, energy saving, environmental pollution is small.
3, the present invention uses supercritical CO2Extracting method prepare Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts,
Semen Platycladi extract, Fructus lycii P.E and Wild jujube seeds extract improve supercritical CO by controlling extracting pressure and temperature2
Fluid is to the dissolubility of active constituents of medicine, to improve recovery rate and purity；It completes to make CO by decompression after extracting2Gasification,
Not residual solvent impurity is conducive to subsequent purification；In extraction process active constituents of medicine is improved using ethyl alcohol as entrainer
Recovery rate, and ethyl alcohol is easily isolated, and is not had any impact to subsequent purification；Without using organic molten in extraction process of the present invention
Agent not only reduces extraction cost, more environmentally protective, safe and efficient.
Specific implementation mode
With reference to embodiment, the following further describes the technical solution of the present invention, and however, it is not limited to this, every right
Technical solution of the present invention is modified or replaced equivalently, and without departing from the spirit of the technical scheme of the invention and range, should all be contained
It covers in protection scope of the present invention.
Embodiment 1
A kind of ginseng oral liquid of invigorating qi for tranquilization of the present embodiment, includes the component of following parts by weight：Ginseng total glycosides extractive 5
~10 parts, 10~30 parts of Astragalus Root P.E, 10~30 parts of siberian Ginseng P.E, 5~20 parts of tuckahoe extracts, Semen Platycladi extract
5~20 parts, 5~20 parts of Fructus lycii P.E, 5~20 parts of Wild jujube seeds extract, small molecule ginseng polypeptide-chelates of zinc 30~80
Part；500 parts of deionized water；
Steps are as follows for the preparation method of its small molecular ginseng polypeptide-chelates of zinc：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, it is that neutral Tris-HCl is buffered that pH value, which is added, by certain mass volume ratio
Solution, after stirring evenly under certain temperature stirring and leaching, collect leaching liquor；It repeats to extract primary and will close leaching liquor twice
And the ultrafiltration membrane that molecular cut off is 5~30KDa is placed it in by after leaching liquor filtering clarification with 0.22~0.45 μm of filter membrane
Device carries out ultrafiltration, and it is the ginseng water-soluble protein crude extract for removing small molecular weight impurity to collect ultrafiltration to retain liquid；
S2, enzymolysis ginseng water-soluble protein prepare small molecule ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5, alkali protease is added, carries out at a certain temperature by ginseng water-soluble protein concentrate
It digests, NaOH solution maintenance system pH value is added dropwise in enzymolysis process, HCl is added by system pH in enzymolysis into enzymatic hydrolysis system after completing
5~7 are adjusted to, neutral proteinase is added, enzymatic hydrolysis system temperature is risen to 90~100 DEG C after completing enzymolysis at a certain temperature and is protected
Holding 10 minutes makes enzyme inactivate, and it is neutrality that pH value is adjusted after cooled to room temperature, obtains enzymolysis liquid；It will using SephadexG-25
The enzymolysis protein classification obtained in enzymolysis liquid, collects the small molecule ginseng polypeptide that molecular weight is 500~1000Da；
S3, small molecule ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected is uniformly mixed with zinc sulfate by certain mass ratio, by reactant
System is adjusted to certain pH value, and the chelating of small molecule ginseng polypeptide and zinc sulfate, water-bath certain time are carried out under certain bath temperature
The ethanol solution that mass concentration is 75% is added in backward system, stands 20~30min, 4000r/min centrifuges 15min, collects
Precipitation, vacuum drying obtain small molecule ginseng polypeptide-chelates of zinc.
Embodiment 2
The present embodiment is differed only in embodiment 1：The ginseng oral liquid of 2 invigorating qi for tranquilization of embodiment includes following weight
The component of part：8 parts of ginseng total glycosides extractive, 20 parts of Astragalus Root P.E, 20 parts of siberian Ginseng P.E, 15 parts of tuckahoe extracts, cypress
15 parts of sub- benevolence extract, 15 parts of Fructus lycii P.E, 15 parts of Wild jujube seeds extract, small molecule ginseng polypeptide -60 parts of chelates of zinc；
500 parts of deionized water.
Embodiment 3
The present embodiment and embodiment 1 difference lies in：The preparation method of 3 small molecule ginseng polypeptide of embodiment-chelates of zinc
Steps are as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, is 1g by mass volume ratio:It is neutral that pH value, which is added, in 10~15mL
The Tris-HCl buffer solutions of 0.01mol/L, after stirring evenly at 4 DEG C 3~6h of stirring and leaching, collect leaching liquor；It repeats to soak
It carries primary and leaching liquor will merge twice, with 0.22~0.45 μm of filter membrane by after leaching liquor filtering clarification, place it in retention point
Son is measured carries out ultrafiltration for the ultra-filtration membrane device of 5~30KDa, and the ginseng that collection ultrafiltration retains liquid as removal small molecular weight impurity is water-soluble
Property protein crude extract；
S2, enzymolysis ginseng water-soluble protein prepare ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5 by ginseng water-soluble protein concentrate, and temperature is 55~70 DEG C, and substrate quality 0.2 is added
~0.6% alkali protease is digested, and NaOH solution maintenance system pH value is added dropwise in enzymolysis process, and 1~2h of enzymolysis is backward
HCl is added in enzymatic hydrolysis system, system pH is adjusted to 5~7, temperature is 50~60 DEG C, and the neutrality of substrate quality 2.0~3.0% is added
Protease is digested, and enzymatic hydrolysis system temperature is risen to 90~100 DEG C after digesting 1~2h and keeps making enzyme inactivate in 10 minutes, from
It is neutrality that pH value is adjusted after being so cooled to room temperature, and obtains enzymolysis liquid；The enzymolysis that will be obtained in enzymolysis liquid using SephadexG-25
Albumen is classified, and collects the small molecule ginseng polypeptide that molecular weight is 500~1000Da；
S3, ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected and zinc sulfate in mass ratio 5~7:1 is uniformly mixed, and adjusts body
Be pH value be 6.5~7.5,50~60 DEG C at a temperature of 30~40min of water-bath, into system be added mass concentration be 75% ethyl alcohol
Solution, standing 20~30min makes Complexes In Solution fully generate, and 4000r/min centrifuges 15min, collects precipitation, vacuum drying
Obtain ginseng polypeptide-chelates of zinc.
Embodiment 4
The present embodiment and embodiment 3 difference lies in：The preparation method of 4 small molecule ginseng polypeptide of embodiment-chelates of zinc
Steps are as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, is 1g by mass volume ratio:It is neutral that pH value, which is added, in 12mL
The Tris-HCl buffer solutions of 0.01mol/L, after stirring evenly at 4 DEG C stirring and leaching 4h, collect leaching liquor；Repeat extraction one
It is secondary and merge leaching liquor twice, with 0.22~0.45 μm of filter membrane by after leaching liquor filtering clarification, place it in molecular cut off
Ultrafiltration is carried out for the ultra-filtration membrane device of 5~30KDa, it is the ginseng water-soluble egg for removing small molecular weight impurity to collect ultrafiltration to retain liquid
White crude extract；
S2, enzymolysis ginseng water-soluble protein prepare ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5 by ginseng water-soluble protein concentrate, and temperature is 60 DEG C, and substrate quality 0.3% is added
Alkali protease is digested, and NaOH solution maintenance system pH value is added dropwise in enzymolysis process, is added into enzymatic hydrolysis system after digesting 1h
Enter HCl and system pH is adjusted to 5~7, temperature is 50 DEG C, and the neutral proteinase that substrate quality 2.0% is added is digested, and digests 1h
Enzymatic hydrolysis system temperature is risen to 90~100 DEG C afterwards and keeps making enzyme inactivate in 10 minutes, pH value is adjusted after cooled to room temperature is
Neutrality obtains enzymolysis liquid；The enzymolysis protein obtained in enzymolysis liquid is classified using SephadexG-25, it is 500 to collect molecular weight
The small molecule ginseng polypeptide of~1000Da；
S3, ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected and zinc sulfate in mass ratio 5:1 is uniformly mixed, regulation system
PH value is water-bath 30min at a temperature of 6.5,50 DEG C, the ethanol solution that addition mass concentration is 75% into system, standing 20~
30min makes Complexes In Solution fully generate, and 4000r/min centrifuges 15min, collects precipitation, and vacuum drying obtains ginseng polypeptide-
Chelates of zinc.
Embodiment 5
The present embodiment and embodiment 3 difference lies in：The preparation method of 5 small molecule ginseng polypeptide of embodiment-chelates of zinc
Steps are as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, is 1g by mass volume ratio:It is neutral that pH value, which is added, in 13mL
The Tris-HCl buffer solutions of 0.01mol/L, after stirring evenly at 4 DEG C stirring and leaching 5h, collect leaching liquor；Repeat extraction one
It is secondary and merge leaching liquor twice, with 0.22~0.45 μm of filter membrane by after leaching liquor filtering clarification, place it in molecular cut off
Ultrafiltration is carried out for the ultra-filtration membrane device of 5~30KDa, it is the ginseng water-soluble egg for removing small molecular weight impurity to collect ultrafiltration to retain liquid
White crude extract；
S2, enzymolysis ginseng water-soluble protein prepare ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5 by ginseng water-soluble protein concentrate, and temperature is 65 DEG C, and substrate quality 0.4% is added
Alkali protease is digested, dropwise addition NaOH solution maintenance system pH value in enzymolysis process, after enzymolysis 1.5h into enzymatic hydrolysis system
HCl is added, system pH is adjusted to 5~7, temperature is 55 DEG C, and the neutral proteinase that substrate quality 2.5% is added is digested, and is digested
Enzymatic hydrolysis system temperature is risen to 90~100 DEG C after 1.5h and keeps making enzyme inactivate in 10 minutes, pH is adjusted after cooled to room temperature
Value is neutrality, obtains enzymolysis liquid；The enzymolysis protein obtained in enzymolysis liquid is classified using SephadexG-25, collecting molecular weight is
The small molecule ginseng polypeptide of 500~1000Da；
S3, ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected and zinc sulfate in mass ratio 6:1 is uniformly mixed, regulation system
PH value is water-bath 35min at a temperature of 7.0,55 DEG C, the ethanol solution that addition mass concentration is 75% into system, standing 20~
30min makes Complexes In Solution fully generate, and 4000r/min centrifuges 15min, collects precipitation, and vacuum drying obtains ginseng polypeptide-
Chelates of zinc.
Embodiment 6
The present embodiment and embodiment 3 difference lies in：The preparation method of 6 small molecule ginseng polypeptide of embodiment-chelates of zinc
Steps are as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, is 1g by mass volume ratio:It is neutral that pH value, which is added, in 14mL
The Tris-HCl buffer solutions of 0.01mol/L, after stirring evenly at 4 DEG C stirring and leaching 6h, collect leaching liquor；Repeat extraction one
It is secondary and merge leaching liquor twice, with 0.22~0.45 μm of filter membrane by after leaching liquor filtering clarification, place it in molecular cut off
Ultrafiltration is carried out for the ultra-filtration membrane device of 5~30KDa, it is the ginseng water-soluble egg for removing small molecular weight impurity to collect ultrafiltration to retain liquid
White crude extract；
S2, enzymolysis ginseng water-soluble protein prepare ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains
Concentrate pH value is adjusted to 8.5~9.5 by ginseng water-soluble protein concentrate, and temperature is 70 DEG C, and substrate quality 0.5% is added
Alkali protease is digested, and NaOH solution maintenance system pH value is added dropwise in enzymolysis process, is added into enzymatic hydrolysis system after digesting 2h
Enter HCl and system pH is adjusted to 5~7, temperature is 60 DEG C, and the neutral proteinase that substrate quality 3.0% is added is digested, and digests 2h
Enzymatic hydrolysis system temperature is risen to 90~100 DEG C afterwards and keeps making enzyme inactivate in 10 minutes, pH value is adjusted after cooled to room temperature is
Neutrality obtains enzymolysis liquid；The enzymolysis protein obtained in enzymolysis liquid is classified using SephadexG-25, it is 500 to collect molecular weight
The small molecule ginseng polypeptide of~1000Da；
S3, ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected and zinc sulfate in mass ratio 7:1 is uniformly mixed, regulation system
PH value is water-bath 40min at a temperature of 7.5,60 DEG C, the ethanol solution that addition mass concentration is 75% into system, standing 20~
30min makes Complexes In Solution fully generate, and 4000r/min centrifuges 15min, collects precipitation, and vacuum drying obtains ginseng polypeptide-
Chelates of zinc.
Embodiment 7
The present embodiment is the preparation method of the ginseng oral liquid of invigorating qi for tranquilization disclosed in embodiment 1-7, is as follows：
Step 1: preparing ginseng total glycosides extractive：
In order to reduce oral solution cost of the present invention, the present embodiment has chosen the total glycosides content height of ginseng, but price is with respect to ginseng
The cheap fresh ginseng flower bud of root and rhizome is raw material, places it in 60~65 DEG C of vacuum drying chambers and does after cleaning up
Ginseng flower bud after abundant drying is crushed to 50~100 mesh by dry 10~15h；It is 1g by mass volume ratio：15~20mL is to institute
It is 40% ethanol solution to obtain and mass concentration is added in ginseng flower bud powder, impregnates 2~3h under room temperature, will be immersed with ginseng flower bud powder
The ethanol solution at end is fitted into the polybag of pressure resistance, is put into after sealing in the extraction vessel of high pressure extraction device；10 under room temperature
The pressure in high pressure extraction device is increased to 300~600Mpa in~15min, it will in 10~20s after 15~25min of pressurize
Pressure in high pressure extraction device is let out rapidly as normal pressure；It repeats after high pressure extraction operates 3~5 times, by polybag from high pressure extraction
It is taken out in the extraction vessel of device, material filtering is removed into filter residue, filtrate is placed under 3000~5000rmp centrifugal rotational speeds and is centrifuged
10~30min, takes supernatant repeated centrifugation 1 time, and supernatant is the total glycosides crude extract of ginseng；The total glycosides crude extract of ginseng is carried out dense
Contracting is adjusted to the concentrate pH value to make it through macroporous absorbent resin D101 chromatographic columns after neutrality and adsorb；It is by mass concentration
20% ethanol solution pH value is adjusted to neutrality and is eluted as the 1st eluting solvent；The ethyl alcohol for being 60% by mass concentration
Solution ph is adjusted to neutrality and is eluted as the 2nd eluting solvent, collects the fraction containing the total glycosides ingredient of ginseng, and decompression is dense
Contracting, spray drying obtain ginseng total glycosides extractive after purification, and the wherein total glycosides purity of ginseng is 90~96%；
Step 2: using supercritical CO2Extracting method prepares Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, cypress
Sub- benevolence extract, Fructus lycii P.E and Wild jujube seeds extract；Prepared Astragalus Root P.E, siberian Ginseng P.E, Poria cocos carry
It is 80~85% to take the mass concentration of object, Semen Platycladi extract, Fructus lycii P.E and Wild jujube seeds extract；
It is carried Step 3: weighing ginseng total glycosides extractive, Astragalus Root P.E, siberian Ginseng P.E, Poria cocos by weight respectively
Object, Semen Platycladi extract, Fructus lycii P.E, Wild jujube seeds extract and small molecule ginseng polypeptide-chelates of zinc are taken, is added 500
Part water, agitated filtering and impurity removing after mixing, filling, high-temperature sterilization obtain the ginseng oral liquid of the invigorating qi for tranquilization.
Embodiment 8
Difference lies in embodiment 8 extracts Radix Astragali, wilsonii, Poria cocos, the seed of Oriental arborvitae, the fruit of Chinese wolfberry to the present embodiment with embodiment 7
With the supercritical CO of spina date seed2Extracting method is：By fully dry Radix Astragali, wilsonii, Poria cocos, the seed of Oriental arborvitae, the fruit of Chinese wolfberry or acid
Jujube kernel raw material crushes, and the Chinese medicine material after taking 100g to crush is put into 1L extraction kettles and is extracted, supercritical CO2Fluid extraction operates
Flow is liquid CO2→ high-pressure pump → extraction kettle (entering supercriticality) → I → separating still of separating still II → cycle；It extracted
CO in journey2Fluid flow is 20L/h, and extraction kettle temperature is 45 DEG C, extracting pressure 30MPa, separating still I, separating still II pressure
Power is 6MPa, and separation temperature is room temperature, uses absolute ethyl alcohol as entrainer in extraction process, and absolute ethyl alcohol dosage is
500mL extracts 2~3h and concentrates gained extract liquor to get Radix Astragali, wilsonii, Poria cocos, the seed of Oriental arborvitae, the fruit of Chinese wolfberry or spina date seed
Extract；Gained Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, Semen Platycladi extract, Fructus lycii P.E and wild jujube
The mass concentration of benevolence extract is 80~85%.
Comparative example 1
This comparative example is differed only in embodiment 1, and the oral liquid prescription of comparative example 1 includes the group of following parts by weight
Point：10~30 parts of Astragalus Root P.E, 10~30 parts of siberian Ginseng P.E, 5~20 parts of tuckahoe extracts, Semen Platycladi extract 5~
20 parts, 5~20 parts of Fructus lycii P.E, 5~20 parts of Wild jujube seeds extract, 500 parts of deionized water.
Comparative example 2
This comparative example is differed only in embodiment 1, and the oral liquid prescription of comparative example 2 only includes the group of following parts by weight
Point：5~10 parts of ginseng total glycosides extractive, small molecule ginseng polypeptide -30~80 parts of chelates of zinc, 500 parts of deionized water.
The clinical test of pesticide effectiveness
Test object：Selection 30~50 years old, there is the man of apathetic, with a haggard face, insomnia forgetfulness, neurasthenia symptom
Property, each 150 of women；
Test method：300 test objects are randomly divided into 5 groups, every group of 30 males, 30 women；
The 1st group of ginseng qi-tonifying Anshen oral liquid that the offer of embodiment 1 is provided,
The 2nd group of ginseng qi-tonifying Anshen oral liquid that the offer of embodiment 2 is provided,
The 3rd group of oral solution for taking the preparation of comparative example 1, the 4th group of oral solution for taking the preparation of comparative example 2,
5th group is taken placebo, and placebo is allocated by caramel colorant and bitters, is free of any drug ingedient；,
Five set takes in the morning and evening daily, adheres to after taking 30 days, apathetic to test object, face is thin and pallid
Worried, insomnia forgetfulness, neurasthenia symptom are investigated, and the results are shown in Table 1：
Table 1
By 1 test result of table it is found that thering is 95.85% or so taking the embodiment of the present invention in the 1st, 2 group of test object
3, after 1 month, apathetic, with a haggard face, insomnia forgetfulness and neurasthenia obtain 4, the 5 invigorating qi for tranquilization ginseng oral liquids prepared
To being effectively relieved；It takes for 3rd group and is only prepared by Radix Astragali, wilsonii, Poria cocos, the seed of Oriental arborvitae, the fruit of Chinese wolfberry and Wild jujube seeds extract
Oral solution and the 4th group of survey for taking the oral solution only prepared by ginseng total glycosides extractive and small molecule ginseng polypeptide-chelates of zinc
It tries in object, only 32.5% and 43% apathetic, with a haggard face, insomnia forgetfulness and neurasthenia symptom are eased；And
Only 3% symptom is alleviated in the 5th group of test object for taking placebo.It can be seen that invigorating qi for tranquilization ginseng mouth of the present invention
Take each active drug ingredient in liquid it is compatible can more effectively play tonifying Qi of reposing, tranquilizing the mind prevent prolapse, reinforce the spleen to benefit the lung, enrich blood it is foster
The effect of the moon, sperm adding and marrow healthing, calming heart brain tonic, to preferably alleviate apathetic user, with a haggard face, insomnia forgetfulness and god
Through weak symptom.
Claims (8)
1. a kind of ginseng oral liquid of invigorating qi for tranquilization, it is characterised in that the ginseng oral liquid of the invigorating qi for tranquilization includes following weight
The component of part：5~10 parts of ginseng total glycosides extractive, 10~30 parts of Astragalus Root P.E, 10~30 parts of siberian Ginseng P.E, Poria cocos carry
Take 5~20 parts of object, 5~20 parts of Semen Platycladi extract, 5~20 parts of Fructus lycii P.E, 5~20 parts, small point of Wild jujube seeds extract
30~80 parts of sub- ginseng polypeptide-chelates of zinc；500 parts of deionized water；The preparation side of its small molecular ginseng polypeptide-chelates of zinc
Method step enters as follows：
S1, extraction ginseng water-soluble protein：
It is cut into small pieces after fresh ginseng is cleaned, it is that neutral Tris-HCl bufferings are molten that pH value, which is added, by certain mass volume ratio
Liquid, after stirring evenly under certain temperature stirring and leaching, collect leaching liquor；It repeats to extract primary and leaching liquor will merge twice,
With 0.22~0.45 μm of filter membrane by after leaching liquor filtering clarification, the ultra-filtration membrane device that molecular cut off is 5~30KDa is placed it in
Ultrafiltration is carried out, it is the ginseng water-soluble protein crude extract for removing small molecular weight impurity to collect ultrafiltration to retain liquid；
S2, enzymolysis ginseng water-soluble protein prepare small molecule ginseng polypeptide：
The ginseng water-soluble protein crude extract that step S1 is obtained is concentrated into the 1/5 of original volume, is cooled to room temperature, obtains ginseng
Concentrate pH value is adjusted to 8.5~9.5, alkali protease is added, carries out enzyme at a certain temperature by water-solubility protein concentrate
It solves, NaOH solution maintenance system pH value is added dropwise in enzymolysis process, HCl is added by system pH tune in enzymolysis into enzymatic hydrolysis system after completing
To 5~7, neutral proteinase is added, enzymatic hydrolysis system temperature is risen to 90~100 DEG C after completing enzymolysis at a certain temperature and is kept
Enzyme is set to inactivate within 10 minutes, it is neutrality that pH value is adjusted after cooled to room temperature, obtains enzymolysis liquid；Using SephadexG-25 by enzyme
The enzymolysis protein classification obtained in solution liquid, collects the small molecule ginseng polypeptide that molecular weight is 500~1000Da；
S3, small molecule ginseng polypeptide-chelates of zinc is prepared：
The small molecule ginseng polypeptide that step S2 is collected is uniformly mixed with zinc sulfate by certain mass ratio, by reaction system tune
To certain pH value, the chelating of small molecule ginseng polypeptide and zinc sulfate are carried out under certain bath temperature, water-bath certain time is backward
The ethanol solution that mass concentration is 75% is added in system, stands 20~30min, 4000r/min centrifuges 15min, collects precipitation,
Vacuum drying obtains small molecule ginseng polypeptide-chelates of zinc.
2. a kind of ginseng oral liquid of invigorating qi for tranquilization according to claim 1, it is characterised in that the ginseng of the invigorating qi for tranquilization
Oral solution includes the component of following parts by weight：8 parts of ginseng total glycosides extractive, 20 parts of Astragalus Root P.E, 20 parts of siberian Ginseng P.E,
15 parts of tuckahoe extracts, 15 parts of Semen Platycladi extract, 15 parts of Fructus lycii P.E, 15 parts of Wild jujube seeds extract, small molecule ginseng
60 parts of polypeptide-chelates of zinc；500 parts of deionized water.
3. a kind of ginseng oral liquid of invigorating qi for tranquilization according to claim 1 or claim 2, it is characterised in that in ginseng total glycosides extractive
The total glycosides purity of ginseng is 90~96%, Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, Semen Platycladi extract, the fruit of Chinese wolfberry
The mass concentration of extract and Wild jujube seeds extract is 80~85%.
4. a kind of ginseng oral liquid of invigorating qi for tranquilization according to claim 3, it is characterised in that ginseng raw material described in step S1
Mass volume ratio with Tris-HCl buffer solutions is 1g:10~15mL；The molar concentration of the Tris-HCl buffer solutions is
0.01mol/L；The temperature of the stirring and leaching is 4 DEG C, and the stirring and leaching time is 3~6h.
5. a kind of ginseng oral liquid of invigorating qi for tranquilization according to claim 4, it is characterised in that basic protein described in step S2
The enzyme concentration of enzyme is the 0.2~0.6% of substrate quality, and hydrolysis temperature is 55~70 DEG C, and enzymolysis time is 1~2h；The neutrality
The enzyme concentration of protease is the 2.0~3.0% of substrate quality, and hydrolysis temperature is 50~60 DEG C, and enzymolysis time is 1~2h.
6. a kind of ginseng oral liquid of invigorating qi for tranquilization according to claim 5, it is characterised in that small molecule people described in step S3
The mass ratio for joining polypeptide and zinc sulfate is 5~7:1；The pH value is 6.5~7.5, and bath temperature is 50~60 DEG C, water bath time
For 30~40min.
7. a kind of preparation method of the ginseng oral liquid of the invigorating qi for tranquilization as described in claim 1-6 any claims, feature
It is that steps are as follows for the preparation method：
Step 1: preparing ginseng total glycosides extractive：
Fresh ginseng flower bud is chosen, dry 10~15h in 60~65 DEG C of vacuum drying chambers is placed it in after cleaning up, will be filled
The ginseng flower bud after drying is divided to be crushed to 50~100 mesh；It is 1g by mass volume ratio：15~20mL is to gained ginseng flower bud powder
Middle addition mass concentration is 40% ethanol solution, impregnates 2~3h under room temperature, will be filled immersed with the ethanol solution of ginseng flower bud powder
In the polybag for entering pressure resistance, it is put into after sealing in the extraction vessel of high pressure extraction device；By high pressure in 10~15min under room temperature
Pressure in extraction element is increased to 300~600Mpa, will be in high pressure extraction device in 10~20s after 15~25min of pressurize
Pressure let out rapidly as normal pressure；It repeats after high pressure extraction operates 3~5 times, by polybag from the extraction vessel of high pressure extraction device
Material filtering is removed filter residue by middle taking-up, and filtrate is placed in 10~30min of centrifugation under 3000~5000rmp centrifugal rotational speeds, is taken
Clear liquid repeated centrifugation 1 time, supernatant are the total glycosides crude extract of ginseng；The total glycosides crude extract of ginseng is concentrated, by concentrate pH
Value, which is adjusted to make it through macroporous absorbent resin D101 chromatographic columns after neutrality, to be adsorbed；The ethanol solution for being 20% by mass concentration
PH value is adjusted to neutrality and is eluted as the 1st eluting solvent；During the ethanol solution pH value that mass concentration is 60% is adjusted to
Property and eluted as the 2nd eluting solvent, collect the fraction containing the total glycosides ingredient of ginseng, be concentrated under reduced pressure, spray drying, obtain
To ginseng total glycosides extractive after purification；
Step 2: using supercritical CO2Extracting method prepares Astragalus Root P.E, siberian Ginseng P.E, tuckahoe extracts, the seed of Oriental arborvitae
Extract, Fructus lycii P.E and Wild jujube seeds extract；
Step 3: weighing 5~10 parts of ginseng total glycosides extractive, 10~30 parts of Astragalus Root P.E, thorn five by following parts by weight respectively
Add 10~30 parts of extract, 5~20 parts of tuckahoe extracts, 5~20 parts of Semen Platycladi extract, 5~20 parts of Fructus lycii P.E, acid
500 parts of water are added in 5~20 parts of jujube kernel extract and small molecule ginseng polypeptide -30~80 parts of chelates of zinc, agitated to be uniformly mixed
Filtering and impurity removing, filling, high-temperature sterilization afterwards, obtain the ginseng oral liquid of the invigorating qi for tranquilization.
8. a kind of preparation method of the ginseng oral liquid of invigorating qi for tranquilization according to claim 7, it is characterised in that step 2 institute
State supercritical CO2Extracting method is：Fully dry Chinese medicine material is crushed, the Chinese medicine material after taking 100g to crush is put into 1L extractions
Kettle is taken to be extracted, supercritical CO2Fluid extraction operating process is liquid CO2→ high-pressure pump → extraction kettle (enters supercritical state
State) → I → separating still of separating still II → cycle；CO in extraction process2Fluid flow is 20L/h, and extraction kettle temperature is 45 DEG C, extraction
Pressure power be 30MPa, separating still I, separating still II pressure be 6MPa, separation temperature is room temperature, is used in extraction process
Absolute ethyl alcohol is as entrainer, and absolute ethyl alcohol dosage is 500mL, and 2~3h is up to Chinese medical extract for extraction.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112841393A (en) * | 2020-12-30 | 2021-05-28 | 广州合诚实业有限公司 | Walnut peptide stable to zinc ions as well as preparation method and application thereof |
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