CN106074682A - Total alkaloid of sophora alopecuroide extract and preparation method thereof and the application prepared in medicament for resisting cervical cancer - Google Patents
Total alkaloid of sophora alopecuroide extract and preparation method thereof and the application prepared in medicament for resisting cervical cancer Download PDFInfo
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Abstract
The present invention relates to Herba Sophorae alopecuroidis technical field, be a kind of total alkaloid of sophora alopecuroide extract and preparation method thereof and the application preparing in medicament for resisting cervical cancer；This total alkaloid of sophora alopecuroide extract obtains as follows: the first step, pulverizes；Second step, reflux, extract,；3rd step, concentrates；4th step, purification.Total alkaloid of sophora alopecuroide extract preparation process of the present invention is simple, need not add organic solvent, and nodules of Sophora alopecuroides from Xinjiang aboundresources, thus greatly reduce production cost in preparation process；Present invention firstly provides total alkaloid of sophora alopecuroide extract as the application preparing in medicament for resisting cervical cancer simultaneously, pharmacy in vitro experiment proves, total alkaloid of sophora alopecuroide extract of the present invention significantly inhibits effect to the growth of HeLa cell, and internal pharmacodynamic experiment shows, it is to mice U14Cervix uteri carcinoma has a certain degree of inhibitory action, illustrates that total alkaloid of sophora alopecuroide extract of the present invention has certain anti-cervical cancer effect.
Description
Technical field
The present invention relates to Herba Sophorae alopecuroidis technical field, be a kind of total alkaloid of sophora alopecuroide extract and preparation method thereof and preparation
Application in medicament for resisting cervical cancer.
Background technology
Herba Sophorae alopecuroidis (Sophora alopecuroides L.) is pulse family (Leguminosae sp.) sophora plant, is one
Plant herbaceos perennial, Uygur entitled " Bu Ya ".Herba Sophorae alopecuroidis Herb bitter in the mouth, cold in nature, it is medicinal include root, stem, herb and
Its seed, has the effects such as heat-clearing and toxic substances removing, dispeiling pathogenic wind and removing dampness, pain relieving parasite killing.Modern pharmacology proves, Herba Sophorae alopecuroidis has following
The pharmacological action of aspect: analgesic activity, antiinflammatory action, antiarrhythmic effect, antiviral；Calmness, antipyretic, cooling, blood fat reducing；Right
Neural effect, hepatoprotective effect, to blood circulation, the effect of respiratory system；Immunoregulation effect；Positive inotropic action,
Blood pressure lowering, reduces heart rate, to myocardial ischemia and the protective effect of infraction；Effect to smooth muscle；Antitumor action；Antiallergic is with flat
Breathe heavily effect.Along with deepening continuously of modern pharmacology pharmacodynamic study, the antitumor of total alkaloid of sophora alopecuroide is worth by the world of medicine
Recognized by.Recent study report shows, the different alkaloid components in Herba Sophorae alopecuroidis are to people's gastric cancer cell line MGC-803, colon cancer
SW620 cell, human erythroleukemia cell strain K562, hepatocellular carcinoma H22, Lewis lung cancer, transitional cell bladder carcinoma cell line, carcinoma of prostate are thin
The tumor cells such as born of the same parents all have inhibited proliferation；Through retrieval, up to the present do not find that Alkaloids From Sophora Alopecuroides L is applied to anti-cervix uteri
Patent in terms of cancer and report.Meanwhile, the technique being currently used for preparing Alkaloids From Sophora Alopecuroides L needs the extraction of very complicated, divides
From, purifying process, use the harmful organic reagents such as chloroform as Extraction solvent, be not suitable for industrialized production.
Summary of the invention
The invention provides a kind of total alkaloid of sophora alopecuroide extract and preparation method thereof and prepare in medicament for resisting cervical cancer
Application, overcome the deficiency of above-mentioned prior art, its preparation technology that can effectively solve existing Alkaloids From Sophora Alopecuroides L is loaded down with trivial details multiple
Miscellaneous and Extraction solvent is chloroform, the problem not being suitable for industrialized production；Present invention firstly provides total alkaloid of sophora alopecuroide simultaneously
Extract is as the application prepared in medicament for resisting cervical cancer.
One of technical scheme is realized by following measures: a kind of total alkaloid of sophora alopecuroide extract,
Obtain as follows: the first step, pulverize, after taking appropriate Herba Sophorae alopecuroidis pulverizing medicinal materials, obtain Herba Sophorae alopecuroidis medicinal material coarse powder；Second step, returns
Stream extracts, and adds reflux, extract, 2 times to 3 times after ethanol water boils in Herba Sophorae alopecuroidis medicinal material coarse powder, and after extraction, sucking filtration obtains
Mixed extract is obtained after filtrate, merging filtrate mix homogeneously；3rd step, concentrates, and obtains concentrated solution after mixed extract is concentrated；4th
Step, purification, macroporous resin first soaks with ethanol water, and the macroporous resin dress post after soaking obtains macroporous resin column,
With the deionized water of 2 times to 5 times column volumes, macroporous resin column is rinsed, after deionized water rinsing, in macroporous resin column
Adding 1.5 times of concentrated solutions to 3 times of column volumes to adsorb, the loading speed of concentrated solution is 0.5 times to 3 times cylinder per hour
Long-pending, adsorption time is 4h to 10h, washes macroporous resin column with the deionized water of 2 times to 5 times column volumes after concentrated solution absorption
De-, then carry out eluting with the ethanol water that percent by volume is 30% to 60% of 3 times to 6 times column volumes, ethanol is water-soluble
The elution rate of liquid is 0.5 times to 5 times column volume per hour, collects ethanol elution, ethanol elution after ethanol water eluting
Liquid is concentrated and dried, obtains the total alkaloid of sophora alopecuroide extract of powder.
Further optimization and/or improvements to foregoing invention technical scheme one of are presented herein below:
In above-mentioned second step, rise to 12 liters of ethanol waters, at Herba Sophorae alopecuroidis by 1 kilogram of Herba Sophorae alopecuroidis medicinal material coarse powder adds 6
Medicinal material coarse powder adds ethanol water and boils rear reflux, extract, 2 times to 3 times, each reflux, extract, 0.5h to 1.5h, ethanol water
Solution be percent by volume be the ethanol water of 50% to 95%.
In above-mentioned 3rd step, mixed extract concentrates and decompression recycling ethanol at temperature is 54 DEG C to 75 DEG C, and mixed extract concentrates
To the 1/16 to 5/24 of ethanol water cumulative volume added during reflux, extract, obtain concentrated solution.
In above-mentioned 4th step, macroporous resin first soaks 20h extremely with the ethanol water that percent by volume is 95% to 98%
36h, the macroporous resin dress post after soaking obtains macroporous resin column；Or/and, in the 4th step, ethanol elution is concentrated to cream
Shape thing, after paste is dried 10h to 14h at temperature is 100 DEG C to 110 DEG C, the total alkaloid of sophora alopecuroide obtaining powder carries
Take thing；Or/and, macroporous resin be HPD400 macroporous resin or HPD700 macroporous resin or HPD500 macroporous resin or HPD750 big
Hole resin.
The two of technical scheme are realized by following measures: a kind of total alkaloid of sophora alopecuroide extract
Preparation method, is carried out: the first step in the steps below, pulverizes, obtains Herba Sophorae alopecuroidis medicinal material coarse powder after taking appropriate Herba Sophorae alopecuroidis pulverizing medicinal materials；
Second step, reflux, extract, Herba Sophorae alopecuroidis medicinal material coarse powder adds reflux, extract, 2 times to 3 times after ethanol water boils, after extraction
Sucking filtration obtains mixed extract after obtaining filtrate, merging filtrate mix homogeneously；3rd step, concentrates, and obtains dense after mixed extract is concentrated
Contracting liquid；4th step, purification, macroporous resin first soaks with ethanol water, and the macroporous resin dress post after soaking obtains greatly
Hole resin column, is rinsed macroporous resin column with the deionized water of 2 times to 5 times column volumes, after deionized water rinsing, at macropore
In resin column add 1.5 times of concentrated solutions to 3 times of column volumes adsorb, the loading speed of concentrated solution be per hour 0.5 times to 3
Times column volume, adsorption time is 4h to 10h, with the deionized water of 2 times to 5 times column volumes to macroporous resin column after concentrated solution absorption
Carry out eluting, then carry out eluting, second with the ethanol water that percent by volume is 30% to 60% of 3 times to 6 times column volumes
The elution rate of alcohol-water solution is 0.5 times to 5 times column volume per hour, collects ethanol elution, second after ethanol water eluting
Alcohol eluen is concentrated and dried, obtains the total alkaloid of sophora alopecuroide extract of powder.
Be presented herein below to foregoing invention technical scheme two further optimization and/or improvements:
In above-mentioned second step, rise to 12 liters of ethanol waters, at Herba Sophorae alopecuroidis by 1 kilogram of Herba Sophorae alopecuroidis medicinal material coarse powder adds 6
Medicinal material coarse powder adds ethanol water and boils rear reflux, extract, 2 times to 3 times, each reflux, extract, 0.5h to 1.5h, ethanol water
Solution be percent by volume be the ethanol water of 50% to 95%.
In above-mentioned 3rd step, mixed extract concentrates and decompression recycling ethanol at temperature is 54 DEG C to 75 DEG C, and mixed extract concentrates
To the 1/16 to 5/24 of ethanol water cumulative volume added during reflux, extract, obtain concentrated solution.
In above-mentioned 4th step, macroporous resin first soaks 20h extremely with the ethanol water that percent by volume is 95% to 98%
36h, the macroporous resin dress post after soaking obtains macroporous resin column；Or/and, in the 4th step, ethanol elution is concentrated to cream
Shape thing, after paste is dried 10h to 14h at temperature is 100 DEG C to 110 DEG C, the total alkaloid of sophora alopecuroide obtaining powder carries
Take thing；Or/and, macroporous resin be HPD400 macroporous resin or HPD700 macroporous resin or HPD500 macroporous resin or HPD750 big
Hole resin.
The three of technical scheme are realized by following measures: a kind of total alkaloid of sophora alopecuroide extract is made
For preparing the application in medicament for resisting cervical cancer.
Total alkaloid of sophora alopecuroide extract preparation process of the present invention is simple, need not add organic solvent in preparation process,
And nodules of Sophora alopecuroides from Xinjiang aboundresources, thus greatly reduce production cost；Present invention firstly provides total alkaloid of sophora alopecuroide simultaneously
Extract is as the application prepared in medicament for resisting cervical cancer, and pharmacy in vitro is it is demonstrated experimentally that total alkaloid of sophora alopecuroide of the present invention extracts
Thing significantly inhibits effect to the growth of HeLa cell, and internal pharmacodynamic experiment shows, U14 in mice cervix uteri carcinoma is had necessarily by it
The inhibitory action of degree, illustrates that total alkaloid of sophora alopecuroide extract of the present invention has certain anti-cervical cancer effect.
Accompanying drawing explanation
Fig. 1 be variable concentrations total alkaloid of sophora alopecuroide extract effect 24h of the present invention after HeLa cell survival rate.
Fig. 2 (a) is apoptosis situation map after matched group HeLa cell 24h.
Fig. 2 (b) is experimental group administration concentration of the present invention apoptosis situation map after HeLa cell 24h when being 6.25mg/ml.
Fig. 2 (c) is experimental group administration concentration of the present invention apoptosis situation map after HeLa cell 24h when being 12.50mg/ml.
Detailed description of the invention
Embodiment 1, this total alkaloid of sophora alopecuroide extract is obtained by following preparation method: the first step, pulverizes, takes appropriate hardship
Herba Sophorae alopecuroidis medicinal material coarse powder is obtained after bean or pea pulverizing medicinal materials；Second step, reflux, extract, Herba Sophorae alopecuroidis medicinal material coarse powder adds ethanol water
Solution boils rear reflux, extract, 2 times to 3 times, and after extraction, sucking filtration obtains mixed extract after obtaining filtrate, merging filtrate mix homogeneously；
3rd step, concentrates, and obtains concentrated solution after mixed extract is concentrated；4th step, purification, macroporous resin first soaks with ethanol water
Bubble, the macroporous resin dress post after soaking obtains macroporous resin column, with the deionized water of 2 times to 5 times column volumes to macroporous resin
Post is rinsed, and after deionized water rinsing, adds 1.5 times of concentrated solutions to 3 times of column volumes and adsorb in macroporous resin column,
The loading speed of concentrated solution is 0.5 times to 3 times column volume per hour, and adsorption time is 4h to 10h, with 2 times after concentrated solution absorption
Deionized water to 5 times of column volumes carries out eluting to macroporous resin column, then by the percent by volume of 3 times to 6 times column volumes is
The ethanol water of 30% to 60% carries out eluting, and the elution rate of ethanol water is 0.5 times to 5 times column volume per hour,
Collecting ethanol elution after ethanol water eluting, ethanol elution is concentrated and dried, and the Herba Sophorae alopecuroidis obtaining powder is total
Alkaloid extract.In pharmacopeia, coarse powder refers to all by No. two sieves, but to be mixed with the powder that can be less than 40% by No. four sieves；
No. two sieves: 24 mesh, No. four sieves: 65 mesh；In 4th step, with the deionized water of 2 times to 5 times column volumes, macroporous resin column is rushed
Washing, after deionized water rinsing, the deionization of outflow is without alcohol taste, and clarification is without muddy；Xinjiang is one of Herba Sophorae alopecuroidis main producing region, the whole nation,
Available natural resources enriches, it is estimated that full boundary Herba Sophorae alopecuroidis total storage is more than 100,000 tons.Xinjiang is multi-national gathering ground,
Uighur "bilingual students" cervical cancer prevalence and mortality rate are significantly higher than this area Han nationality and other ethnic groups, there are the obvious people
Race's feature difference.The present invention benefits the people for making full use of area resources of medicinal plant advantage, plays traditional national medicine anti-
Particular utility in terms of tumor.
Embodiment 2, this total alkaloid of sophora alopecuroide extract is obtained by following preparation method: the first step, pulverizes, takes appropriate hardship
Herba Sophorae alopecuroidis medicinal material coarse powder is obtained after bean or pea pulverizing medicinal materials；Second step, reflux, extract, Herba Sophorae alopecuroidis medicinal material coarse powder adds ethanol water
Solution boils rear reflux, extract, 2 times or 3 times, and after extraction, sucking filtration obtains mixed extract after obtaining filtrate, merging filtrate mix homogeneously；
3rd step, concentrates, and obtains concentrated solution after mixed extract is concentrated；4th step, purification, macroporous resin first soaks with ethanol water
Bubble, the macroporous resin dress post after soaking obtains macroporous resin column, with the deionized water of 2 times or 5 times column volumes to macroporous resin
Post is rinsed, and after deionized water rinsing, the concentrated solution adding 1.5 times or 3 times column volumes in macroporous resin column adsorbs,
The loading speed of concentrated solution is 0.5 times or 3 times of column volumes per hour, and adsorption time is 4h or 10h, with 2 times after concentrated solution absorption
Or the deionized water of 5 times of column volumes carries out eluting to macroporous resin column, then by the percent by volume of 3 times or 6 times column volumes it is
The ethanol water of 30% or 60% carries out eluting, and the elution rate of ethanol water is 0.5 times or 5 times of column volumes per hour,
Collecting ethanol elution after ethanol water eluting, ethanol elution is concentrated and dried, and the Herba Sophorae alopecuroidis obtaining powder is total
Alkaloid extract.
Embodiment 3, as the optimization of above-described embodiment, in second step, adds 6 liters by 1 kilogram of Herba Sophorae alopecuroidis medicinal material coarse powder
To 12 liters of ethanol waters, Herba Sophorae alopecuroidis medicinal material coarse powder adds reflux, extract, 2 times to 3 times after ethanol water boils, every time
Reflux, extract, 0.5h to 1.5h, ethanol water be percent by volume be the ethanol water of 50% to 95%.
Embodiment 4, as the optimization of above-described embodiment, in the 3rd step, mixed extract concentrates at temperature is 54 DEG C to 75 DEG C
And decompression recycling ethanol, after mixed extract is concentrated into during reflux, extract, the 1/16 to 5/24 of added ethanol water cumulative volume, obtain
Concentrated solution.
Embodiment 5, as the optimization of above-described embodiment, in the 4th step, macroporous resin first with percent by volume be 95% to
The ethanol water of 98% soaks 20h to 36h, and the macroporous resin dress post after soaking obtains macroporous resin column；Or/and, the 4th
In step, ethanol elution is concentrated to paste, after paste is dried 10h to 14h at temperature is 100 DEG C to 110 DEG C, obtains
The total alkaloid of sophora alopecuroide extract of powder；Or/and, macroporous resin be HPD400 macroporous resin or HPD700 macroporous resin or
HPD500 macroporous resin or HPD750 macroporous resin.
Embodiment 6, this total alkaloid of sophora alopecuroide extract is as the application prepared in medicament for resisting cervical cancer.
The total alkaloid of sophora alopecuroide extract that above-described embodiment 1 to embodiment 5 obtains is as preparing in medicament for resisting cervical cancer
Application, effect experiment is as follows:
One. total alkaloid of sophora alopecuroide extract In Vitro Anti cervical cancer effect of the present invention
1. material
0.25% pancreatin, hyclone, DMEM culture fluid, the detection examination of CCK-8, Annexin V-FITC/PI apoptosis
Agent box, human cervical carcinoma Hela cell's strain.
2. method
Exponential phase HeLa cell is inoculated in 96 orifice plates (100 μ l/ hole), is placed in 5%CO2, in 37 DEG C of incubators,
After cultivating the growth of 24h cell attachment, experimental group is separately added into 6.25mg/ml, 8.75mg/ml, 12.50mg/ml variable concentrations
The total alkaloid of sophora alopecuroide solution of extract 100 μ l that the embodiment of the present invention 1 to embodiment 5 obtains, is all provided with 6 multiple holes；Comparison
It is 5000 Cell sap 100 μ l that group adds cell concentration, and after continuing to cultivate 24h, every hole adds 10%CCK-8 and cell is cultivated mixed
Closing solution 100 μ l, continue to hatch 3h, use microplate reader to measure absorption value at 450nm, calculate cell survival rate, formula is as follows:
Cell survival rate=(ODExperiment-ODComparison)/(ODComparison-ODComparison) × 100%
Take the logarithm the HeLa cell of trophophase, obtain single cell suspension through trypsinization, centrifugal (1100rpm, 4min), collect
Cell.Adjust cell density with the DMEM culture fluid containing 10%FBS, add 12 orifice plates with 1 × 105/hole, 1ml/ hole, 37
DEG C, 5%CO2 cell culture incubator cultivate 24h.Arranging 3 matched groups, experimental group is separately added into 6.25mg/ml, 12.50mg/ml not
The total alkaloid of sophora alopecuroide solution of extract 100 μ l obtained with the embodiment of the present invention 1 to the embodiment 5 of concentration, 37 DEG C, 5%
After CO2 cell culture incubator cultivates 24h, collect each group of cell with 5mlEP pipe.First, drawing cell culture fluid, PBS washs 1 time,
Collect cleaning mixture, add 0.5ml trypsin digestion cell, add 1ml culture fluid and blow and beat back and forth 20 times, collect culture fluid, centrifugal
(1100rpm, 4min), sucks supernatant, adds 1mlPBS washing, and this process is repeated 2 times, and adds and combines liquid 500 μ l mix homogeneously,
Piping and druming is uniformly.Room temperature lucifuge operates, and adds Annexin V-FITC 5 μ l mix homogeneously, then adds PI5 μ l mix homogeneously, room temperature lucifuge
Dyeing 15min；Cross 300 eye mesh screens, immediately flow cytometer detection.
3. result
Use CCK-8 method to measure and add 6.25mg/ml, 8.75mg/ml, 12.50mg/ml variable concentrations embodiment of the present invention
The survival rate of HeLa cell after the total alkaloid of sophora alopecuroide solution of extract effect 24h that 1 to embodiment 5 obtains.With matched group
Undressed HeLa cell is compared, along with the increase of total alkaloid of sophora alopecuroide solution of extract administration concentration of the present invention,
In identical action time, cell survival rate is in being decreased obviously trend.Total alkaloid of sophora alopecuroide solution of extract of the present invention is given
When concentration is 8.75mg/ml, the HeLa cell death of more than 50% can be made.Result shows, total alkaloid of sophora alopecuroide of the present invention
Extract significantly inhibits effect to the growth of HeLa cell.The total alkaloid of sophora alopecuroide extract effect of the present invention of variable concentrations
After 24h, HeLa cell survival rate is shown in Fig. 1.
After matched group HeLa cell 24h, apoptosis situation is shown in Fig. 2 (a)；When experimental group administration concentration of the present invention is 6.25mg/ml
After HeLa cell 24h, apoptosis situation is shown in Fig. 2 (b)；When experimental group administration concentration of the present invention is 12.50mg/ml after HeLa cell 24h
Apoptosis situation is shown in Fig. 2 (c)；From Fig. 2 (a), Fig. 2 (b) and Fig. 2 (c) it can be seen that addition 6.25mg/ml, 12.50mg/ml is different
After the total alkaloid of sophora alopecuroide solution of extract that the concentration embodiment of the present invention 1 to embodiment 5 obtains processes 24h, use streaming
Cell art detection apoptosis, experimental group apoptosis rate is followed successively by 16.0%, 33.3%, hence it is evident that higher than matched group, and along with
The increase of administration concentration, early apoptosis rate and the late apoptic rate of cell the most substantially increase.
Two. anti-cervical cancer effect in total alkaloid of sophora alopecuroide extract body of the present invention
1. material
Animal: kunming mice, female, body weight 20 ± 2g, Beijing Vital River Experimental Animals Technology Co., Ltd. provide.
Cell strain: U14 in mice cervix uteri carcinoma strain (ascitic type), is provided by institute of Materia Medica,Chinese Academy of Medical Sciences.
2. method
Taking 40 female KM mices, certain density U14 tumor liquid is inoculated in right fore oxter, and 0.2ml/ is only.Next day mice
Weigh, be randomly divided into 4 groups, 10/group, respectively blank group (saline control group) (Control), positive drug
(CTX) matched group, total alkaloid of sophora alopecuroide extract low dose group (50mg/kg) of the present invention, total alkaloid of sophora alopecuroide of the present invention
Extract high dose group (100mg/kg).After inoculation 24h, positive drug (CTX) matched group intraperitoneal injection of cyclophosphamide 1 time.This
Bright total alkaloid of sophora alopecuroide extract low dose group and high dose group gastric infusion, successive administration 10 days.During experiment, every day is seen
Examine the situations such as the general activity of mice, fur, feces.After the last 24h of administration, disconnected neck is put to death, and weighs and tumor weight, and counts
Calculate tumour inhibiting rate.
3. result
Total alkaloid of sophora alopecuroide extract low dose group of the present invention and high dose group have certain journey to U14 in mice cervix uteri carcinoma
The inhibitory action of degree, illustrates that total alkaloid of sophora alopecuroide extract of the present invention has a certain degree of suppression to U14 in mice cervix uteri carcinoma
Effect, the results are shown in Table 1.
Tumour inhibiting rate (%)=[(blank group average tumor weight-administration group average tumor weight)/blank group average tumor weight)
× 100%
In sum, total alkaloid of sophora alopecuroide extract preparation process of the present invention is simple, need not add in preparation process
Organic solvent, and nodules of Sophora alopecuroides from Xinjiang aboundresources, thus greatly reduce production cost；Present invention firstly provides Herba Sophorae alopecuroidis simultaneously
Total alkaloids extract is as the application prepared in medicament for resisting cervical cancer, and pharmacy in vitro is it is demonstrated experimentally that Herba Sophorae alopecuroidis of the present invention is always given birth to
Alkaloids extract significantly inhibits effect to the growth of HeLa cell, and internal pharmacodynamic experiment shows, it is to U14 in mice cervical cancer
Tumor has a certain degree of inhibitory action, illustrates that total alkaloid of sophora alopecuroide extract of the present invention has certain anti-cervical cancer effect.
Above technical characteristic constitutes embodiments of the invention, and it has stronger adaptability and implementation result, can basis
It is actually needed the non-essential technical characteristic of increase and decrease, meets the demand of different situations.
Table 1 Alkaloids From Sophora Alopecuroides L of the present invention extract is to U14The tumor-inhibiting action of subcutaneous tumor-bearing mice (n=10,)
Note: compared with Control group, * * P ＜ 0.01.
Claims (9)
1. a total alkaloid of sophora alopecuroide extract, it is characterised in that obtain as follows: the first step, pulverizes, takes appropriate hardship
Herba Sophorae alopecuroidis medicinal material coarse powder is obtained after bean or pea pulverizing medicinal materials；Second step, reflux, extract, Herba Sophorae alopecuroidis medicinal material coarse powder adds ethanol water
Solution boils rear reflux, extract, 2 times to 3 times, and after extraction, sucking filtration obtains mixed extract after obtaining filtrate, merging filtrate mix homogeneously；
3rd step, concentrates, and obtains concentrated solution after mixed extract is concentrated；4th step, purification, macroporous resin first soaks with ethanol water
Bubble, the macroporous resin dress post after soaking obtains macroporous resin column, with the deionized water of 2 times to 5 times column volumes to macroporous resin
Post is rinsed, and after deionized water rinsing, adds 1.5 times of concentrated solutions to 3 times of column volumes and adsorb in macroporous resin column,
The loading speed of concentrated solution is 0.5 times to 3 times column volume per hour, and adsorption time is 4h to 10h, with 2 times after concentrated solution absorption
Deionized water to 5 times of column volumes carries out eluting to macroporous resin column, then by the percent by volume of 3 times to 6 times column volumes is
The ethanol water of 30% to 60% carries out eluting, and the elution rate of ethanol water is 0.5 times to 5 times column volume, second per hour
Collecting ethanol elution after alcohol-water solution eluting, ethanol elution is concentrated and dried, and the Herba Sophorae alopecuroidis obtaining powder is always given birth to
Alkaloids extract.
Total alkaloid of sophora alopecuroide extract the most according to claim 1, it is characterised in that in second step, by 1 kilogram of bitter bean
Sub-medicinal material coarse powder adds 6 and rises to 12 liters of ethanol waters, add in Herba Sophorae alopecuroidis medicinal material coarse powder after ethanol water boils and return
Stream extract 2 times to 3 times, each reflux, extract, 0.5h to 1.5h, ethanol water be percent by volume be the ethanol of 50% to 95%
Aqueous solution.
Total alkaloid of sophora alopecuroide extract the most according to claim 1 and 2, it is characterised in that in the 3rd step, mixed extract exists
Temperature is to concentrate and decompression recycling ethanol at 54 DEG C to 75 DEG C, and it is overall that mixed extract is concentrated into added ethanol water during reflux, extract,
Behind long-pending 1/16 to 5/24, obtain concentrated solution.
4. according to the total alkaloid of sophora alopecuroide extract described in claim 1 or 2 or 3, it is characterised in that in the 4th step, macropore tree
Fat first soaks 20h to 36h with the ethanol water that percent by volume is 95% to 98%, and the macroporous resin dress post after soaking obtains
Macroporous resin column；Or/and, in the 4th step, ethanol elution is concentrated to paste, and paste is 100 DEG C to 110 DEG C in temperature
Under be dried 10h to 14h after, obtain the total alkaloid of sophora alopecuroide extract of powder；Or/and, macroporous resin is HPD400 macropore
Resin or HPD700 macroporous resin or HPD500 macroporous resin or HPD750 macroporous resin.
5. the preparation method of a total alkaloid of sophora alopecuroide extract, it is characterised in that carry out in the steps below: the first step, powder
Broken, obtain Herba Sophorae alopecuroidis medicinal material coarse powder after taking appropriate Herba Sophorae alopecuroidis pulverizing medicinal materials；Second step, reflux, extract, in Herba Sophorae alopecuroidis medicinal material coarse powder
Middle addition ethanol water boils rear reflux, extract, 2 times to 3 times, and after extraction, sucking filtration obtains filtrate, merging filtrate mix homogeneously
After obtain mixed extract；3rd step, concentrates, and obtains concentrated solution after mixed extract is concentrated；4th step, purification, macroporous resin first uses second
Alcohol-water solution soaks, will soak after macroporous resin dress post obtain macroporous resin column, with 2 times to 5 times column volumes go from
Macroporous resin column is rinsed by sub-water, after deionized water rinsing, adds 1.5 times to 3 times of column volumes in macroporous resin column
Concentrated solution adsorbs, and the loading speed of concentrated solution is 0.5 times to 3 times column volume per hour, and adsorption time is 4h to 10h, dense
Macroporous resin column is carried out eluting, then with 3 times to 6 times column volumes with the deionized water of 2 times to 5 times column volumes after the absorption of contracting liquid
The ethanol water that percent by volume is 30% to 60% carry out eluting, the elution rate of ethanol water is 0.5 times per hour
To 5 times of column volumes, collecting ethanol elution after ethanol water eluting, ethanol elution is concentrated and dried, obtains powder
The total alkaloid of sophora alopecuroide extract of shape.
The preparation method of total alkaloid of sophora alopecuroide extract the most according to claim 5, it is characterised in that in second step, press
1 kilogram of Herba Sophorae alopecuroidis medicinal material coarse powder adds 6 and rises to 12 liters of ethanol waters, Herba Sophorae alopecuroidis medicinal material coarse powder adds ethanol water
Boil rear reflux, extract, 2 times to 3 times, each reflux, extract, 0.5h to 1.5h, ethanol water be percent by volume be 50% to
The ethanol water of 95%.
7. according to the preparation method of the total alkaloid of sophora alopecuroide extract described in claim 5 or 6, it is characterised in that the 3rd step
In, mixed extract concentrates and decompression recycling ethanol at temperature is 54 DEG C to 75 DEG C, and mixed extract is concentrated into added second during reflux, extract,
Behind the 1/16 to 5/24 of alcohol-water solution cumulative volume, obtain concentrated solution.
8. according to the preparation method of the total alkaloid of sophora alopecuroide extract described in claim 5 or 6 or 7, it is characterised in that the 4th
In step, macroporous resin first soaks 20h to 36h with the ethanol water that percent by volume is 95% to 98%, the macropore after soaking
Resin dress post obtains macroporous resin column；Or/and, in the 4th step, ethanol elution is concentrated to paste, and paste in temperature is
After being dried 10h to 14h at 100 DEG C to 110 DEG C, obtain the total alkaloid of sophora alopecuroide extract of powder；Or/and, macroporous resin
For HPD400 macroporous resin or HPD700 macroporous resin or HPD500 macroporous resin or HPD750 macroporous resin.
9. one kind according to the total alkaloid of sophora alopecuroide extract described in claim 1 or 2 or 3 or 4 as preparing anti-cervical cancer medicine
Application in thing.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107488187A (en) * | 2017-03-21 | 2017-12-19 | 南方医科大学 | Applications of the Sophora alopecuroide alkali dimer A ~ D in anti-inflammatory or anti-tumor medicinal preparation is prepared |
| CN112402506A (en) * | 2019-08-22 | 2021-02-26 | 张爱华 | Traditional Chinese medicine composition for inhibiting HPV virus and/or treating cervical cancer precancerous lesion |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1275571A (en) * | 1999-06-22 | 2000-12-06 | 内蒙古苦参生化集团有限责任公司 | Process for preparing oxymatrine from sophora alopecuroide |
| CN101129455A (en) * | 2007-09-18 | 2008-02-27 | 新疆维吾尔自治区药物研究所 | Sophora extractive and method of preparing the same and application of the same |
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2016
- 2016-06-27 CN CN201610480276.3A patent/CN106074682A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1275571A (en) * | 1999-06-22 | 2000-12-06 | 内蒙古苦参生化集团有限责任公司 | Process for preparing oxymatrine from sophora alopecuroide |
| CN101129455A (en) * | 2007-09-18 | 2008-02-27 | 新疆维吾尔自治区药物研究所 | Sophora extractive and method of preparing the same and application of the same |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107488187A (en) * | 2017-03-21 | 2017-12-19 | 南方医科大学 | Applications of the Sophora alopecuroide alkali dimer A ~ D in anti-inflammatory or anti-tumor medicinal preparation is prepared |
| CN112402506A (en) * | 2019-08-22 | 2021-02-26 | 张爱华 | Traditional Chinese medicine composition for inhibiting HPV virus and/or treating cervical cancer precancerous lesion |
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