Summary of the invention
In order to overcome the deficiencies in the prior art and shortcoming, primary and foremost purpose of the present invention is the preparation method providing a kind of Semen Ziziphi Spinosae extract.
Another object of the present invention is to the Semen Ziziphi Spinosae extract providing above-mentioned preparation method to prepare.
Another object of the present invention is to provide a kind of brain-strengthening sleeping oral liquid containing above-mentioned Semen Ziziphi Spinosae extract.
Object of the present invention is achieved through the following technical solutions:
A preparation method for Semen Ziziphi Spinosae extract, comprises following steps:
(1) Semen Ziziphi Spinosae is pulverized, and obtains spiny jujuba seed powder; Spiny jujuba seed powder hot water extraction's active substance, filters, obtains filtrate 1 and filtering residue 1;
(2) acid solution of filtering residue 1 pH 1 ~ 5 that step (1) prepares soaks, and filters, obtains filtrate 2 and filtering residue 2; Filtering residue 2 soaks with the alkaline solution of pH 8 ~ 10 again, filters, obtains filtrate 3 and filtering residue 3;
(3) filtrate 1,2 and 3 for preparing of combining step (1) and step (2), concentrated, obtain concentrated solution;
(4) the concentrated solution ethanol prepared in step (3) is carried out precipitate with ethanol, except impurity such as destarching, phlegmatic temperament, natural gum, protein, oils and fats, fat-soluble pigment, polysaccharide, inorganic salt, tannins; Centrifugal, collect filtrate, obtain Semen Ziziphi Spinosae extract;
The temperature of the hot water extraction described in step (1) is preferably 75 ~ 80 DEG C, and the time of hot water extraction is preferably 2 ~ 4h;
The time that acid solution described in step (2) soaks is preferably 2 ~ 4h;
The pH value of the acid solution described in step (2) is preferably 5;
Acid solution described in step (2) is preferably hydrochloric acid solution;
The time that alkaline solution described in step (2) soaks is preferably 2 ~ 4h;
The pH value of the alkaline solution described in step (2) is preferably 8.2;
Alkaline solution described in step (2) is preferably sodium hydroxide solution;
Concentrated preferably concentrating under reduced pressure or freeze concentration described in step (3);
The volume fraction of the ethanol described in step (4) is preferably 75% ~ 80%;
The time of the precipitate with ethanol described in step (4) is preferably 3 ~ 4h;
A kind of Semen Ziziphi Spinosae extract, is prepared by above-mentioned preparation method;
Described Semen Ziziphi Spinosae extract comprises 8 kinds of Flavonoid substances, Semen Ziziphi Spinosae alkaloid, jujuboside class material; (Fig. 1 ~ 4)
8 kinds of described Flavonoid substances are respectively swertisin (in Fig. 1 formula I), versulin-6-C-β-D-glucopyanoside (in Fig. 1 formula II), puerarin (in Fig. 1 formula III), 6 " '-ferulic acid Ziziphi Spinosae flavone c-glycoside (in Fig. 1 formula IV), Ziziphi Spinosae flavone carbon glycoside glycosides (Fig. 1 Chinese style V), isovitexin (in Fig. 1 VI formula), different Ziziphi Spinosae flavone c-glycoside (Fig. 1 Chinese style VII) and 6 " ' the different Ziziphi Spinosae flavone c-glycoside of-ferulic acid (Fig. 1 Chinese style VIII);
The alkaloidal structural formula of described Semen Ziziphi Spinosae as shown in Figure 2;
Described jujuboside class material is Semen Ziziphi Spinosae saponin D and Semen Ziziphi Spinosae saponin E; (Fig. 3 and Fig. 4)
In described Semen Ziziphi Spinosae extract, the content of Flavonoid substances is 0.576 ~ 0.594mg/g, and the content of jujuboside class material is 0.365 ~ 0.382mg/g;
A brain-strengthening sleeping oral liquid containing Semen Ziziphi Spinosae extract, comprises following component by mass percentage:
Above-mentioned Semen Ziziphi Spinosae extract 20% ~ 22%;
Mel 8% ~ 10%;
Water supplies 100%;
The preparation method of the described brain-strengthening sleeping oral liquid containing Semen Ziziphi Spinosae extract, comprises following steps:
Each component of the above-mentioned brain-strengthening sleeping oral liquid containing Semen Ziziphi Spinosae extract hooked and adjust, sterilizing, obtains the brain-strengthening sleeping oral liquid containing Semen Ziziphi Spinosae extract;
Sterilizing 25min at the condition of described sterilizing is preferably 78 ~ 80 DEG C;
Have the mainly two class materials of syngignoscism of calming the nerves in Semen Ziziphi Spinosae, i.e. Flavonoid substances and Semen Ziziphi Spinosae saponin class material, concrete structure is shown in Fig. 1, Fig. 2, Fig. 3 and Fig. 4.The present invention carries out abstraction and purification to the active substance in Semen Ziziphi Spinosae with syngignoscism of calming the nerves, obtain Semen Ziziphi Spinosae extract, further science is mixed with brain-strengthening sleeping oral liquid, and by measuring the active and toxicological effect of perfusion mice righting reflex loss and the mode Test extraction thing of recovery time, proof Semen Ziziphi Spinosae extract has no side effect, determined curative effect.This brain-strengthening sleeping oral liquid may be used for the symptom such as insomnia, anxiety because brain overwork, psychentonia, operating pressure cause greatly, have calm the nerves, brain-strengthening, hypnosis, the effect such as Fructus Alpiniae Oxyphyllae.
The present invention has following advantage and effect relative to prior art:
(1) preparation method of Semen Ziziphi Spinosae extract provided by the invention is simple to operation: by Semen Ziziphi Spinosae pulverizing, active substance water extraction, semi-bionic extraction, concentrated, alcohol deposition method decontamination, obtain Semen Ziziphi Spinosae extract; Effective ingredient retains many; The organic solvent that whole leaching process relates to only has ethanol, and safety is controlled, has the advantage to human non-toxic's evil.
(2) yield of product that the preparation method of Semen Ziziphi Spinosae extract provided by the invention obtains is significantly improved.
(3) the brain-strengthening sleeping oral liquid containing Semen Ziziphi Spinosae extract provided by the invention contains Semen Ziziphi Spinosae sleep aiding and nerve soothing effective ingredient, comprise Semen Ziziphi Spinosae flavonoid substance, Semen Ziziphi Spinosae alkaloid, Semen Ziziphi Spinosae saponin D and E, have no side effect, tranquillizing and allaying excitement determined curative effect, may be used for the symptoms such as the insomnia because brain overwork, psychentonia, operating pressure cause greatly, anxiety.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
(1) with the pulverizer through sterilization, the Semen Ziziphi Spinosae after air-dry or airing is pulverized, obtain spiny jujuba seed powder; Spiny jujuba seed powder is with the hot-water soak of 80 DEG C after 3 hours, and filter, obtain filtrate 1 and filtering residue 1, wherein, filtrate 1 cryopreservation is for subsequent use;
(2) semi-bionic extraction: the filtering residue 1 that step (1) the prepares hydrochloric acid solution of pH 5 soaks, filter after 3 hours, obtain filtrate 2 and filtering residue 2, wherein, filtrate 2 cryopreservation is for subsequent use; Filtering residue 2 filters after soaking 3 hours with the sodium hydroxide solution of pH8.2, and obtain filtrate 3 and filtering residue 3, wherein, filtrate 3 cryopreservation is for subsequent use;
(3) concentrated: the filtrate 1,2 and 3 that combining step (1) and step (2) prepare, adopt reduction vaporization to concentrate, obtain concentrated solution;
(4) precipitate with ethanol: the ethanol being 80% by the concentrated solution volume fraction that step (3) prepares carries out precipitate with ethanol, except impurity such as destarching, phlegmatic temperament, natural gum, protein, oils and fats, fat-soluble pigment, polysaccharide, inorganic salt, tannins.After precipitate with ethanol carries out 3 hours, collected by centrifugation filtrate, obtains Ziziphi Spinosae extract, and extraction ratio is 4%.
Embodiment 2
(1) with the pulverizer through sterilization, the Semen Ziziphi Spinosae after air-dry or airing is pulverized, obtain spiny jujuba seed powder; Spiny jujuba seed powder is with the hot-water soak of 75 DEG C after 4 hours, and filter, obtain filtrate 1 and filtering residue 1, wherein, filtrate 1 cryopreservation is for subsequent use;
(2) semi-bionic extraction: the filtering residue 1 that step (1) the prepares hydrochloric acid solution of pH 1 soaks, filter after 2 hours, obtain filtrate 2 and filtering residue 2, wherein, filtrate 2 cryopreservation is for subsequent use; Filtering residue 2 filters after soaking 2 hours with the sodium hydroxide solution of pH10, and obtain filtrate 3 and filtering residue 3, wherein, filtrate 3 cryopreservation is for subsequent use;
(3) concentrated: the filtrate 1,2 and 3 that combining step (1) and step (2) prepare, adopt reduction vaporization to concentrate, obtain concentrated solution;
(4) precipitate with ethanol: the ethanol being 75% by the concentrated solution volume fraction that step (3) prepares carries out precipitate with ethanol, except impurity such as destarching, phlegmatic temperament, natural gum, protein, oils and fats, fat-soluble pigment, polysaccharide, inorganic salt, tannins.After precipitate with ethanol carries out 3.5 hours, collected by centrifugation filtrate, obtains Ziziphi Spinosae extract, and extraction ratio is 4%.
Embodiment 3
(1) with the pulverizer through sterilization, the Semen Ziziphi Spinosae after air-dry or airing is pulverized, obtain spiny jujuba seed powder; Spiny jujuba seed powder is with the hot-water soak of 78 DEG C after 2 hours, and filter, obtain filtrate 1 and filtering residue 1, wherein, filtrate 1 cryopreservation is for subsequent use;
(2) semi-bionic extraction: the filtering residue 1 that step (1) the prepares hydrochloric acid solution of pH 4 soaks, filter after 4 hours, obtain filtrate 2 and filtering residue 2, wherein, filtrate 2 cryopreservation is for subsequent use; Filtering residue 2 filters after soaking 4 hours with the sodium hydroxide solution of pH9, and obtain filtrate 3 and filtering residue 3, wherein, filtrate 3 cryopreservation is for subsequent use;
(3) concentrated: the filtrate 1,2 and 3 that combining step (1) and step (2) prepare, adopt reduction vaporization to concentrate, obtain concentrated solution;
(4) precipitate with ethanol: the ethanol being 78% by the concentrated solution volume fraction that step (3) prepares carries out precipitate with ethanol, except impurity such as destarching, phlegmatic temperament, natural gum, protein, oils and fats, fat-soluble pigment, polysaccharide, inorganic salt, tannins.After precipitate with ethanol carries out 4 hours, collected by centrifugation filtrate, obtains Ziziphi Spinosae extract, and extraction ratio is 4%.
Embodiment 4
(1) jujuboside D in Semen Ziziphi Spinosae extract, E qualitative
Take methanol as solvent, be mixed with the jujuboside D of concentration 1mg/mL and the solution of jujuboside E (all purchased from Nat'l Pharmaceutical & Biological Products Control Institute), in contrast product solution 1 and reference substance solution 2 respectively.
Test according to thin layer chromatography (" Chinese Pharmacopoeia " 2005 editions), draw need testing solution (Semen Ziziphi Spinosae extract that embodiment 1,2 and 3 prepares) and each 10 μ L of blank liquid (methanol), the each 5 μ L of reference substance solution 1,2, put respectively on same silica gel g thin-layer plate, with water saturated butanol solution for developing solvent, launch, take out, dry, spray the vanillin-sulfuric acid solution of 1% (mass percent), inspect immediately.In test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of display same color, thus the effective ingredient demonstrated in the Semen Ziziphi Spinosae extract that embodiment 1,2 and 3 prepares is jujuboside D, E.
(2) quantitative experiment of saponins material in Semen Ziziphi Spinosae extract:
The preparation of standard curve: precision takes ginsenoside Rb1's reference substance (purchased from the Nat'l Pharmaceutical & Biological Products Control Institute) 5.0mg of dry constant weight, moves to 10mL volumetric flask, with dehydrated alcohol standardize solution, is mixed with the reference substance solution of 0.5mg/mL.The general Rb1 reference substance solution (C=0.5mg/mL) 0 of accurate absorption Radix Ginseng soap, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6ml is in 10mL tool plug rub oral examination tube, volatilize reagent in an oven, add 5% (mass percent) vanillin-glacial acetic acid solution 0.2mL and perchloric acid 0.8mL, shake up, 60 DEG C of heating in water bath 20min, 3min is cooled in ice-water bath, add glacial acetic acid 5mL, shake up, retinue reagent is done blank, absorbance is measured at 545nm place with spectrophotometer, with ginsenoside Rb1's concentration for abscissa, absorption angle value is vertical coordinate, drawing standard curve, found that ginsenoside Rb1 within the scope of 0.01-0.05mg/mL in good linear relation, obtain regression equation y=17.925x-0.0498, R
2=0.9988.
The each 0.5mL of Semen Ziziphi Spinosae extract that accurate extraction embodiment 1,2 and 3 prepares, be placed in 10mL tool plug rub oral examination tube, operate according to " preparation of standard curve ", by above-mentioned standard curve, try to achieve the content of saponins material: the content that embodiment 1,2 and 3 prepares saponins material in Semen Ziziphi Spinosae extract is respectively 0.382mg/g, 0.365mg/g, 0.370mg/g.
(3) quantitative experiment of Flavonoid substances in Semen Ziziphi Spinosae extract:
The preparation of standard curve: accurate draw rutin standard solution (purchased from Nat'l Pharmaceutical & Biological Products Control Institute) (C=0.2mg/mL) 0,2,4,6,8,10mL is in the volumetric flask of 50mL, adding distil water is diluted to 10mL, then adds 5% (mass percent) NaNO
2solution 2mL, shakes up, and leaves standstill 6min; Add 10% (mass percent) Al (NO again
3)
3solution 2mL, shakes up, and leaves standstill 6min; Finally add 4% (mass percent) NaOH solution 20mL, be settled to scale with distilled water, shake up, leave standstill 15min, measure absorbance with spectrophotometer at 510nm place.With the concentration of control substance of Rutin for abscissa, absorbance is vertical coordinate, and drawing standard curve obtains regression equation y=13.113x-0.0177, R
2=0.9994.
The accurate each 2mL of Semen Ziziphi Spinosae extract drawing above-described embodiment 1,2 and 3 and prepare, be placed in 50mL volumetric flask, operate according to " preparation of standard curve ", by above-mentioned standard curve, try to achieve the content of Flavonoid substances: embodiment 1,2 and 3 prepares the content of Flavonoid substances in Semen Ziziphi Spinosae extract and is respectively 0.594mg/g, 0.576mg/g, 0.580mg/g.
The preparation of the brain-strengthening sleeping oral liquid of embodiment 5 containing Semen Ziziphi Spinosae extract
(1) tune is hooked: the Semen Ziziphi Spinosae extract, Mel and the water that embodiment 1 are prepared hook adjusts mixing; Wherein, the mass percent of Semen Ziziphi Spinosae extract, Mel and water is respectively 20%, 8%, 72%;
(2) subpackage and sterilizing: step (1) is hooked the mixed liquor after adjusting and carries out pipe dress, often prop up small bottle packing 10 milliliters.Carry out disinfection with pasteurizer, sterilize at 80 DEG C 25min, then carry out bacterial number sanitary index inspection (reference standard is food hygiene quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed casees.
Storage of finished products at low temperature, ventilation, dry place, or is stored in 4 DEG C of refrigerators, and the shelf-life reaches more than 6 months.
The preparation of the brain-strengthening sleeping oral liquid of embodiment 6 containing Semen Ziziphi Spinosae extract
(1) tune is hooked: the Semen Ziziphi Spinosae extract, Mel and the water that embodiment 1 are prepared hook adjusts mixing; Wherein, the mass percent of Semen Ziziphi Spinosae extract, Mel and water is respectively 21%, 9%, 70%;
(2) subpackage and sterilizing: step (1) is hooked the mixed liquor after adjusting and carries out pipe dress, often prop up small bottle packing 10 milliliters.Carry out disinfection with pasteurizer, sterilize at 78 DEG C 24min, then carry out bacterial number sanitary index inspection (reference standard is food hygiene quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed casees.
Storage of finished products at low temperature, ventilation, dry place, or is stored in 4 DEG C of refrigerators, and the shelf-life reaches more than 6 months.
The preparation of the brain-strengthening sleeping oral liquid of embodiment 7 containing Semen Ziziphi Spinosae extract
(1) tune is hooked: the Semen Ziziphi Spinosae extract, Mel and the water that embodiment 1 are prepared hook adjusts mixing; Wherein, the mass percent of Semen Ziziphi Spinosae extract, Mel and water is respectively 22%, 10%, 68%;
(2) subpackage and sterilizing: step (1) is hooked the mixed liquor after adjusting and carries out pipe dress, often prop up small bottle packing 10 milliliters.Carry out disinfection with pasteurizer, sterilize at 79 DEG C 20min, then carry out bacterial number sanitary index inspection (reference standard is food hygiene quality standard), qualified labeling.Every 10 fill a box, every 50 box-packed casees.
Storage of finished products at low temperature, ventilation, dry place, or is stored in 4 DEG C of refrigerators, and the shelf-life reaches more than 6 months.
The direct sleep effect of embodiment 8 Semen Ziziphi Spinosae extract
Mice (purchased from Guangzhou Experimental Animal Center) 40, male and female half and half, are divided into 4 groups at random.Wherein test group 3 groups, namely brain-strengthening sleeping oral liquid high (30mL/kg), in (20mL/kg), low (10mL/kg) dosage group.All the other one group is Normal group, and brain-strengthening sleeping oral liquid is prepared by embodiment 5.Reaching more than 40 seconds for falling asleep with mice righting reflex loss after gavage oral liquid, reverting to righting reflex and reviving, record and calculate number of elements, sleep rate, time for falling asleep and the sleep time of often organizing mice sleep.Result is as shown in table 1.
Result shows, sleep does not appear in Normal group mice, movable normal.Each dosage brain-strengthening sleeping oral liquid all can make mice occur sleep (table 1).
The direct sleep laboratory of table 1 brain-strengthening sleeping oral liquid
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.