CN104666377A - Extraction and purification technology of total triterpenoids in scandent schefflera stem and leaf - Google Patents
Extraction and purification technology of total triterpenoids in scandent schefflera stem and leaf Download PDFInfo
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Abstract
The invention relates to an extraction and purification technology of total triterpenoids in scandent schefflera stem and leaf. The technology comprises the following steps: A taking a scandent schefflera stem and leaf medicinal material, crushing to prepare scandent schefflera stem and leaf powder and putting into a container; B adding ethanol of which the volume concentration is 95% to the container, wherein the solid-liquid rate of the scandent schefflera stem and leaf powder to ethanol is 10g/mL, heating in a water bath, wherein the heating temperature in the water bath is 90 DEG C; C carrying out reflux extract for 60 minutes, and filtering an extract liquid to form a filtrate; and D carrying out vacuum concentration on the filtrate to obtain a concentrate, and drying the concentrate to obtain a crude extract. By virtue of the technology, extraction of total triterpenoids in scandent schefflera stem and leaf is carried out; and the mass fraction of the total triterpenoids in the scandent schefflera stem and leaf crude extract can reach 22.09%. Meanwhile, the invention further provides a purification technology of the total triterpenoids in the scandent schefflera stem and leaf. The crude extract is purified by selecting the optimal process operation conditions, so that the content of the total triterpenoids in the final extract of the scandent schefflera stem and leaf reaches 45.98%.
Description
Technical field
The present invention relates to a kind of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene extraction and purification process.
Background technology
Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) is Araliaceae Schefflera plant Schefflera arboricola Hayata, originates in the ground such as Fujian, Taiwan, Guangdong, Yunnan, is the traditional Chinese herbal medicine among the people of China, containing number of chemical composition, comprises: volatile oil; Organic acid; Triterpenes; Alkaloids; Phenols; Protein-based; Panaxynol etc., have widely pharmacologically active as antiinflammatory, analgesia, tranquilizing soporific and lax bronchial smooth muscle etc. and clinical efficacy, clinically for the treatment such as traumatic injury, rheumatic arthritis.
Commercially available Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) sheet has expelling wind and removing dampness, promoting blood circulation and stopping pain function.At present common anti inflammatory analgesic is chemical synthetic drug clinically, and side effect is large, and some also has serious additive.And Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) preparation comparatively speaking, anti-inflammatory and antalgic is evident in efficacy, and to treatment trigeminal neuralgia instant effect, toxicity is little.But at present Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) preparation is used as medicine mainly with crude extract, the production technology system of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) sheet directly beats powder with Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) crude drug and is used as medicine, and make tablet specification bigger than normal, patient's dose is many, and quality and the drug effect of medicine have greater room for improvement.
Also not about the correlation technique of the Isolation and purification of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene in current document.The Isolation and purification method about total triterpene in other Chinese crude drugs is had in some patent documentations, as Authorization Notice No. CN 101785799 B, name is called the Chinese invention patent of " extract of total triterpenes in dracocephalum moldavica and its production and use ", it discloses total triterpenes in dracocephalum moldavica compound extracting method, comprise the steps:
(1) concentrated by Herba Dracocephali ethanol extract, to reclaim after ethanol to obtain extractum, extractum adds water boiling washing, filters;
(2) extractum dissolves with alkaline solution, filters, gets filtrate;
(3) filtrate adds concentrated hydrochloric acid, stirs, and filters, obtains insoluble matter;
(4) not tolerantly water lotion is eluted in neutral with distilled water;
(5) insoluble matter is with dissolve with ethanol, bleaching agent bleaching;
(6) polycaprolactam resin column on the ethanol after decolouring, with the alcohol flushing chromatographic column not higher than 30%, collects
30% ethanol elution part, concentrated, dry, obtaining final product, the weight percentage of extract obtained middle total triterpene is not less than 50%.
Although the method can be used for extracting the total triterpene of Herba Dracocephali, but during extraction in order to total triterpene in Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae), poor effect.
For another example, application publication number CN 104189021 A, Chinese invention patent application that name is called " a kind of Cinnamomum kanahirai hay bacterium Antrodia Camphorata total triterpene extracting method ", it discloses a kind of Antrodia Camphorata total triterpene extracting method, comprises the following steps:
(1) sample pretreatment: take Antrodia Camphorata 10-20g, dries to constant weight, pulverizes and sieves, and adds 10-20 times of water dissolution, obtained testing sample solution;
(2) the obtained testing sample solution in step (1) is dissolved in solvent, and adds active carbon, filtering with microporous membrane, and it is for subsequent use to get filtrate;
(3) add in the ultrasonic mixing of ether 10-20ml in obtained in step (2) sample filtrate, extract, extract operation repetition 2 times, add n-butyl alcohol 25-40ml, ultrasonic mixing, extract operation and repeat 3-5 time, obtain sample crude extract;
(4) by concentrated, centrifugal for sample crude extract obtained in step (3), standing 10-20min, precipitation is adopted water wash 2-4 time, filters and to obtain precipitation;
(5) precipitation obtained in step (4) added in 50ml measuring bottle, add solvent standardize solution, add sodium bicarbonate solution 5-10ml and extract 3-4 time, centrifuging and taking precipitates, dry must sample total triterpenes material extract.
The method is in order to the extraction of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene, and extraction effect is also unsatisfactory.Also record the extracting method of total triterpene in other Chinese crude drugs in the prior art, but owing to containing various different chemical composition in different Chinese crude drugs, the different reagent added in leaching process and adopt different technological parameters that these chemical compositions all may be made to interact, thus the purity extracted of impact and extraction effect.Total triterpene is subject to the impact of medical material nature in the extraction of different Chinese crude drug.
At present, also do not have the Isolation and purification method about total triterpene in Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae), Given this, inventor furthers investigate the Isolation and purification technique of total triterpene in Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae), then has this case to produce.
Summary of the invention
The object of the present invention is to provide a kind of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene extraction and purification process.
In order to achieve the above object, the present invention adopts such technical scheme:
A kind of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene extraction and purification process, it comprises the following steps:
A, get Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) medical material, pulverize and make Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder, put into container;
B, in this container, add the ethanol that volumetric concentration is 95% again, the solid-liquid ratio of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder and ethanol is 10g/mL, carries out heating in water bath, and water bath heating temperature is 90 DEG C;
C, then carry out reflux, extract, reflux extracting time is 60 minutes, then extracting solution is carried out filtration obtains filtrate;
D, filtrate is carried out concentrating under reduced pressure obtain concentrate, concentrate is carried out drying and obtains crude extract.
E, the crude extract taken in above-mentioned steps D, be configured to the sample solution that concentration is 0.884mg/mL;
F, will the AB-8 macroporous adsorbent resin of dried resin 5g be equivalent to, and load diameter by wet method dress post and be 20mm, highly be the cylinder formation resin column of 300mm, carry out remove impurity with distilled water eluting resin column;
G, by resin column on the sample solution in step e, first use the remove impurity of distilled water eluting, then add volumetric concentration be 80% ethanol carry out eluting, ethanol elution flow velocity is 1BV/h, wherein 1BV=30mL, ethanol elution consumption is 8BV, wherein 1BV=30mL, collect ethanol elution, this eluent evaporate to dryness is obtained finished product.
Adopt Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene extraction process in the present invention, obtain optimised process operating condition by test of many times, with this understanding, in Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) crude extract, the mass fraction of total triterpene can reach 22.09%.; By selecting optimised process operating condition to carry out purification to crude extract, the content of total triterpene in final extract (i.e. finished product) in Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) is made to reach 45.98%.
Accompanying drawing illustrates:
Fig. 1 is total triterpene static adsorption kinetic curve in the present invention;
Fig. 2 is the adsorption isotherm of AB-8 macroporous adsorbent resin in the present invention;
Fig. 3 be in the present invention sample solution concentration on the impact of total triterpene adsorbance;
Fig. 4 be in the present invention different pH on the impact of total triterpene adsorbance;
Fig. 5 be in the present invention AB-8 macroporous adsorbent resin to the adsorption curve of total triterpene;
Fig. 6 is AB-8 macroporous adsorbent resin elution curve in the present invention;
Detailed description of the invention
In order to explain technical scheme of the present invention further, be described in detail below in conjunction with embodiment.
Embodiment one
A, get Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) medical material, pulverize and make Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder, accurately take 9 parts of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder, every part of 20g, put respectively in round-bottomed flask;
B, in this round-bottomed flask, add the ethanol of finite concentration and volume again, carry out heating in water bath, water bath heating temperature is 90 DEG C;
C, then carry out reflux, extract, extracting solution is carried out filtration and obtains filtrate;
D, filtrate is carried out concentrating under reduced pressure obtain concentrate, concentrate is carried out drying and obtains crude extract.
Wherein, Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) is provided by Quanzhou, Fujian Province Pharma Inc. of Zhong Qiao Group Plc, through being accredited as the dry root stem and leaf of S.arboricola Hayata.Concentrating under reduced pressure adopts RE 52-86A type Rotary Evaporators to carry out.
Adopt above-mentioned steps, carry out orthogonal test: choose concentration of alcohol, solid-liquid ratio, extraction time 3 affect the principal element of extract total triterpene amount, each factor establishes 3 levels (see factor level table), carries out L
9(3
3) orthogonal test.
Wherein concentration of alcohol is volumetric concentration;
Solid-liquid ratio is the Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder adding 1g, the volume of the corresponding ethanol added, and unit is g/mL;
The content of total triterpene adopts UV-2450 type ultraviolet-visible spectrophotometer (Japanese Shimadzu Corporation) to measure, and selects vanillin-glacial acetic acid-perchloric acid as the developer of colorimetric method for determining Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene amount.
Factor level table
Tab.Factors and levels
Orthogonal experiments is as follows:
Orthogonal experiments
Tab.Results of orthogonal test
Wherein, K
1, K
2, K
3represent the summation of extract total triterpene contents under each level of each factor, R represents extreme difference.Carry out variance analysis, analysis result is as follows:
The results of analysis of variance
Tab.Results of variance analysis
Note: in variance analysis, F
0.05(2,2)=19.00, * P<0.05.(" * " is the sign of significance in analysis of variance table, represents through this factor P<0.05 of variance analysis, have statistical significance, therefore factor A has a significant impact to result).
The extracted amount of total triterpene and extraction efficiency in comprehensive Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae), determine extraction process of the present invention: adopt concentration to be the ethanol of 95%, solid-liquid ratio is 10g/mL, and reflux extracting time is 60 minutes.
Take a certain amount of resin (the AB-8 macroporous adsorbent resin of Beijing Suo Laibao Science and Technology Ltd.), with 95% ethanol (volumetric concentration) soak 24h fully swelling after, pour out ethanol and floating thing, carry out wet method dress post.Repeatedly rinse with 95% ethanol and mix (with volume ratio 1:4 mixing) with distilled water to eluent and do not occur white casse, then with distilled water balance pillar to resin arrangement evenly, bubble-free, eluent is without alcohol taste, for subsequent use.
Carry out the purification of crude extract as follows further:
E, the crude extract taken in above-mentioned steps D, be configured to the sample solution that concentration is 0.884mg/mL;
F, will the AB-8 macroporous adsorbent resin of dried resin 5g be equivalent to, and load diameter by wet method dress post and be 20mm, highly be the cylinder formation resin column of 300mm, carry out remove impurity with distilled water eluting resin column;
G, by resin column on the sample solution in step e, first use the remove impurity of distilled water eluting, then add volumetric concentration be 80% ethanol carry out eluting, ethanol elution flow velocity is 1BV/h, wherein 1BV=30mL, ethanol elution consumption is 8BV, wherein 1BV=30mL, collect ethanol elution, this eluent evaporate to dryness is obtained finished product.
The present invention determines optimised process particular by following test:
1, the static adsorption of macroporous resin and desorbing
1.1, Static Adsorptive capacity is investigated: accurately take 3 kinds of different dry resin (D101 macroporous adsorbent resins that pretreatment is dried, X-5 macroporous adsorbent resin, AB-8 macroporous adsorbent resin) each 2.0g, be placed in 100mL tool plug triangular flask, add 20mL sample liquid (to be configured by above-mentioned crude extract, total triterpene concentration is 0.884mg/mL), be placed in agitator, agitator adopts the TS-1 type agitator of its woods Bel instrument manufacturing company limited of Haimen City, at room temperature, with 120r/min vibration 24h, filter after abundant absorption, measure the concentration of total triterpene in surplus solution.The adsorbance Q (mg/g) of resin to total triterpene is calculated, formula 1:Q=(C by formula 1
0-C
e) × V/W, the results are shown in Table 1.
1.2, static Performance of resolving adds 95% alcoholic solution 20mL in 3 kinds of saturated macroporous resins of absorption, is placed in agitator, at room temperature, with 120r/min vibration 24h, filters, measures the concentration of total triterpene in 95% alcoholic solution.Total triterpene resolution factor B (%) is calculated, formula 2:B=(C by formula 2
b× V
b/ QW) × 100%, the results are shown in Table 1.
Wherein, in formula 1, formula 2: Q: adsorbance, mg/g; C
0: initial concentration, mg/mL; C
e: residual concentration, mg/mL; V: liquor capacity, mL; W: the quality of dried resin, g; B: desorption efficiency; C
b: the concentration of total triterpene in 95% alcoholic solution, mg/mL; V
b: the volume of 95% alcoholic solution, mL.
Table 1
3 kinds of macroporous resin adsorption amounts and desorption efficiency measurement result
Tab 1The static adsorption capacity and elution ratio by the three types of macroporous resin
As seen from the results in Table 1, compared with D101, X-5 two kinds of non-polar resins, the absorption analysis feature of low pole Resin A B-8 macroporous adsorbent resin to Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene composition is relatively better.
Wherein, D101 macroporous adsorbent resin is provided by Chemical Reagent Co., Ltd., Sinopharm Group.
Wherein, X-5 macroporous adsorbent resin is provided by Cangzhou Bao En Science and Technology Ltd..
1.3, static adsorption kinetic curve: accurately take AB-8 macroporous adsorbent resin dried resin 2.0g, be placed in the tool plug conical flask of 100mL, add sample liquid 20mL, be placed in agitator, room temperature 120r/min vibrates, and measures the concentration of total triterpene in solution at set intervals, calculate the adsorbance Q of resin in t, take time as abscissa, adsorbance is vertical coordinate mapping, the results are shown in Figure 1.
As shown in Figure 1, before 1h, the rate of adsorption is very fast, and along with the prolongation of adsorption time, the rate of adsorption declines, and during 2h, absorption reaches balance substantially, and therefore selected 2h is the static adsorption time, selects the equilibrium adsorption time to be 2h when measuring adsorption isothermal curve.Visible AB-8 macroporous adsorbent resin resin has good adsorption dynamics adsorption kinetics characteristic to Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene, comparatively fast can reach adsorption equilibrium, be suitable for suitability for industrialized production.
1.4, AB-8 absorption with macroporous adsorbent resin isothermal line measures and accurately takes 5 parts of each 2.0g of AB-8 macroporous adsorbent resin dried resin, be placed in the tool plug conical flask of 5 100mL, add the Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene crude extract sample liquid 20mL of 5 variable concentrations respectively, be placed in agitator, room temperature 120r/min vibrates 2h, filter, measure the concentration of total triterpene in solution.According to the relation in adsorbance after adsorption equilibrium and solution between total triterpene concentration, obtain the adsorption isotherm of this resin, abscissa represents equilibrium concentration, and unit is mg/mL, and vertical coordinate represents equilibrium adsorption capacity, and unit mg/g, the results are shown in Figure 2.
The absorption behavior that adsorption isotherm can predict macroporous resin in dynamic adsorption experiment is measured in Staticadsorption experiment.As shown in Figure 2, adsorption isotherm is asymptote, absorption behavior meets Langmuir adsorption isotherm, can think that the absorption of AB-8 macroporous resin to Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) triterpenoid compound is monolayer adsorption, therefore lower sample concentration should be adopted, increase sample solution concentration and can not improve adsorbance greatly, the loss of triterpenoid compound can be caused on the contrary.
2. the dynamic adsorption of macroporous resin and desorbing
2.1, sample solution concentration is on the impact of absorption property: accurately take a certain amount of AB-8 macroporous adsorbent resin (being equivalent to 2.5g dried resin), and wet method dress post (1BV=15mL), 95% ethanol elution, distilled water is eluted to without alcohol taste.By above-mentioned crude extract prepare respectively total triterpene concentration be 0.221,0.442,0.663,0.884, each 30mL of sample solution of 1.105mg/mL, collect effluent, measure total triterpene concentration, calculate adsorbance, the results are shown in Figure 3, wherein abscissa represents sample solution total triterpene concentration, unit mg/mL, vertical coordinate represents adsorbance, unit mg/g.
As shown in Figure 3, along with the increase of sample solution concentration, the adsorbance of AB-8 macroporous adsorbent resin to total triterpene increases gradually, and when concentration is 0.884mg/mL, adsorbance is maximum, and when concentration increases again, adsorbance reduces.Water solublity in conjunction with sample increases with concentration and reduces, and determines that sample solution concentration is advisable with 0.884mg/mL.
2.2, sample solution pH value accurately takes a certain amount of AB-8 macroporous adsorbent resin (being equivalent to 2.5g dried resin) to the impact of absorption property, and wet method dress post (1BV=15mL), 95% ethanol elution, distilled water is eluted to without alcohol taste.Get sample solution (pH value is 5.11) 6 parts of each 90mL, regulate the pH of 5 increment liquid with 1%HCl or 5%NaOH solution, be adjusted to respectively: 3.10,4.11,6.09,7.10,8.11.By 6 increment liquid with flow velocity 1BV/h upper prop respectively, collect effluent, measure total triterpene concentration, calculate adsorbance, the results are shown in Figure 4.
As shown in Figure 4, the adsorbance of AB-8 macroporous adsorbent resin to the total triterpene composition of different pH sample liquid is more or less the same, and the pH of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) sample liquid is little on the impact of macroporous resin adsorption Triterpenoid, therefore selects to carry out column purification at the pH of sample liquid itself.
2.3, the investigation of maximum applied sample amount accurately takes a certain amount of AB-8 macroporous adsorbent resin (being equivalent to 5.0g dried resin), and wet method dress post (1BV=30mL), 95% ethanol elution, distilled water is eluted to without alcohol taste.By 240mL sample solution with flow velocity 1BV/h upper prop, Fractional Collections effluent, every 10mL milliliter collects 1 part, totally 24 parts.Measure effluent total triterpene concentration, be abscissa with effluent volume, in effluent, total triterpene concentration is vertical coordinate, draws dynamic adsorption curve, the results are shown in Figure 5.
As shown in Figure 5, the sample solution upper prop being 0.884mg/mL with total triterpene concentration, the relative concentration sample solution concentration of the effluent total triterpene of front 5BV is in reduced levels, and now resin is better to the adsorption effect of triterpenoid compound, and amount of leakage is less.During upper prop effluent 6BV, effluent total triterpene concentration is 10.05% of sample solution concentration, now reaches leak point, and total triterpene starts to leak, therefore determines that maximum applied sample amount is 180mL.Wherein, the leakage point of macroporous adsorbent resin also deserves to be called post terminal, and when sample solution is constantly added on resin column, resin starts adsorbed target thing, and when sample solution adds to certain volume, resin absorption reaches necessarily saturated, and adsorbance reduces.Detection method is Fractional Collections effluent, measures wherein object content, and draws volume-concentration curve, and the concentration revealing liquid when somewhere is that 10% of the front upper sample liquid concentration value of upper prop is leak point.
2.4,180mL sample solution upper prop (AB-8 macroporous adsorbent resin dry weight 5.0g is got in the investigation of the suitableeest eluant strength, 1BV=30mL), respectively with distilled water, 20%, 40%, 60%, 80%, 95%, the ethanol of 100% carries out eluting, elution flow rate 1BV/h, collect the eluent of different concentration ethanol, water bath method, weighs.Precision takes each eluate 5.00mg, measures total triterpene contents, determines best eluant strength.The results are shown in Table 2.
Table 2 different concentration ethanol is on the impact of resin elution effect
Tab 2Effect of ethanol concentrations on desorption quantities of macroret ieular resis
As seen from the results in Table 2, higher with the total triterpene contents of 60%, 80%, 95% ethanol elution products therefrom, the total triterpene contents of other concentration products therefroms is lower.Select which kind of eluant except the total triterpene contents according to eluted product, simultaneously will in conjunction with eluate amount, although the total triterpene contents of 95% ethanol elution products therefrom is the highest, eluate amount is less, therefore considers, and selects 80% ethanol.
2.5,180mL sample solution upper prop (AB-8 macroporous adsorbent resin dry weight 5.0g is got in the investigation of eluting agent, 1BV=30mL), successively with distilled water eluting resin column, use 80% ethanol-eluting resin column of 10BV again, elution flow rate 1BV/h, collect eluent, every 10mL milliliter collects 1 part, totally 30 parts.Measure eluent total triterpene concentration, the results are shown in Figure abscissa in 6, figure and represent column volume, unit BV, vertical coordinate represents total triterpene concentration, unit mg/mL.
As shown in Figure 6, when 80% ethanol elution consumption is 8BV, the quality of accumulation solid content has reached 99% of total effluent solid quality, component high for the total triterpene contents of more than 98% can be eluted.In conjunction with thin layer chromatography, with 1% vanillin concentrated sulphuric acid ethanol for developer directing terminal, eluting is substantially complete, therefore selected 80% ethanol elution consumption is 8BV.
3. demonstration test: preparation total triterpene concentration is sample liquid 3 parts of each 180mL of 0.884mg/mL, crosses AB-8 macroporous adsorptive resins (AB-8 macroporous adsorbent resin dry weight 5.0g, 1BV=30mL) with 1BV/h flow velocity.First use the remove impurity of distilled water eluting resin column, then use 80% ethanol elution, elution flow rate 1BV/h, elution amount 8BV, collect eluent, water bath method, weighs.Precision takes 3 parts of each 5.00mg of eluate, measures total triterpene mass fraction.Result obtains, and the eluting rate after 3 increment product cross post is respectively 47.43%, 49.22%, 47.78%, and average elution rate is 48.14%; The total triterpene mass fraction of gained 3 parts of eluates is respectively 45.36%, 44.24%, 48.34%, and average mass fraction is 45.98%.
The present invention is by static adsorption and desorption experiment, find that AB-8 macroporous adsorbent resin has good static adsorption, desorption performance to seven leaf total triterpenoids, be applicable to the purification of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene composition, AB-8 purification with macroreticular resin Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene has the features such as selectivity is high, high adsorption capacity, operation is simple.By dynamic adsorption and desorption experiment, determine that the technique of AB-8 purification with macroreticular resin Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene is: the dry AB-8 macroporous adsorbent resin of 5g is loaded diameter be 20mm, highly be 300mm cylinder in, with the sample solution loading under natural pH condition, sample solution concentration is 0.884mg/mL, applied sample amount 6BV, carries out eluting with 80% ethanol of 8BV.Under these these process conditions, the eluting rate of product reaches 48.14%, and the mass fraction of total triterpene reaches 45.98%.
In an embodiment of the present invention, except " static adsorption of macroporous resin and desorbing ", this part tests after resin used is through drying in advance, become dried resin, directly take, other part Experiment resin used is wet resin, and what adopt in experiment is the resin of the water content about 68% of directly buying.
In the present invention, the size of resin bed volume (BV) value and experiment amount of resin used number relevant, " 2.1, sample solution concentration on the impact of absorption property " and " 2.2, sample solution pH value on the impact of absorption property " part in a specific embodiment, accurately take a certain amount of AB-8 macroporous adsorbent resin (being equivalent to 2.5g dried resin), the resin column volume 1BV=15mL of its correspondence, other part resin column volumes 1BV=30mL is then corresponding with a certain amount of AB-8 macroporous adsorbent resin (being equivalent to 5.0g dried resin).
Claims (1)
1. a Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) total triterpene extraction and purification process, is characterized in that: it comprises the following steps:
A, get Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) medical material, pulverize and make Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder, put into container;
B, in said vesse, add the ethanol that volumetric concentration is 95% again, the solid-liquid ratio of Radix Schefflerae Arboricolae (Caulis et Folium Schefflerae Arboricolae) powder and ethanol is 10g/mL, carries out heating in water bath, and water bath heating temperature is 90 DEG C;
C, then carry out reflux, extract, reflux extracting time is 60 minutes, then extracting solution is carried out filtration obtains filtrate;
D, filtrate is carried out concentrating under reduced pressure obtain concentrate, concentrate is carried out drying and obtains crude extract.
E, the crude extract taken in above-mentioned steps D, be configured to the sample solution that concentration is 0.884mg/mL;
F, will the AB-8 macroporous adsorbent resin of dried resin 5g be equivalent to, and load diameter by wet method dress post and be 20mm, highly be the cylinder formation resin column of 300mm, carry out remove impurity with distilled water eluting resin column;
G, by resin column on the sample solution in step e, first use the remove impurity of distilled water eluting, then add volumetric concentration be 80% ethanol carry out eluting, ethanol elution flow velocity is 1BV/h, wherein 1BV=30mL, ethanol elution consumption is 8BV, wherein 1BV=30mL, collect ethanol elution, this eluent evaporate to dryness is obtained finished product.
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| CN101433608A (en) * | 2008-12-11 | 2009-05-20 | 中国人民武装警察部队医学院 | Effective component of radix rosae giganteae in treating diarrhea and preparation method |
| CN103417588A (en) * | 2013-08-23 | 2013-12-04 | 中国科学院新疆理化技术研究所 | Preparation method of effective part of Hippophae rhamnoides |
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| CN101433608A (en) * | 2008-12-11 | 2009-05-20 | 中国人民武装警察部队医学院 | Effective component of radix rosae giganteae in treating diarrhea and preparation method |
| CN103417588A (en) * | 2013-08-23 | 2013-12-04 | 中国科学院新疆理化技术研究所 | Preparation method of effective part of Hippophae rhamnoides |
Non-Patent Citations (1)
| Title |
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| 吕超田等: "灵芝三萜类化合物提取纯化及检测技术研究进展", 《安徽农业科学》 * |
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