CN105198851A - Purification method for blackberry anthocyanin - Google Patents
Purification method for blackberry anthocyanin Download PDFInfo
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- CN105198851A CN105198851A CN201510606373.8A CN201510606373A CN105198851A CN 105198851 A CN105198851 A CN 105198851A CN 201510606373 A CN201510606373 A CN 201510606373A CN 105198851 A CN105198851 A CN 105198851A
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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Abstract
The invention discloses a purification method for blackberry anthocyanin. The purification method includes the steps of resin pretreatment and blackberry anthocyanin crude extract purification. Efficient purification of the blackberry anthocyanin can be achieved, and the purity of the obtained blackberry anthocyanin is high. According to the purification method, polyamide resin is selected as adsorbent and has the advantages that the large specific area and the large granularity are achieved, adsorption and desorption capacity is high, cost is low, regeneration is easy, and a preparation method is simple. Through research on the static and dynamic adsorption and desorption process of the blackberry anthocyanin, purification conditions are optimized, the initial concentration of the blackberry anthocyanin, the pH value of adsorption liquid, the pH value of eluent and the optimal value of ethanol concentration are determined, and the adsorption rate and the elution rate of the blackberry anthocyanin are both high. According to the method, time-saving and efficient purification of the blackberry anthocyanin can be achieved, purification time can be shortened, and cost can be reduced.
Description
Technical field
The present invention relates to a kind of purification process of blackberry, blueberry anthocyanidin, belong to plant extract technical field of purification.
Background technology
The method extracting anthocyanidin from plant is a kind of nonselective extracting method, except pigment, can produce other a large amount of compounds, as carbohydrate, glycitols, organic acid, amino acid, protein etc., will affect the stability of pigment.Therefore, purifying is carried out to the anthocyanidin in blackberry extract and is very important, effectively can promote pigment stability, promote its application in food-processing.In prior art, the method for purifying anthocyanidin is mainly CO
2method, column chromatography, sorbent material method and aqueous two phase extraction technique.
Absorption is a kind of method of simple and effective purifying biological activeconstituents.The sorbent material being used to purifying effective constituent (ball pine tree, flavonoid and polyphenol etc.) from natural phant recently has macroporous adsorbent resin, Zeo-karb and polyamide resin.Wherein, polyamide resin a kind ofly has large specific surface area and the resin of volume particle size.Because polyamide resin sorption and desorption ability is high, cost is low, regenerating easily, preparation method is simple, thus causes and pays close attention to widely.Polyamide resin can remove most impurity from the mixture of complexity, thus improves the content of chemical substance to be purified, is widely used in separating effective ingredient from natural phant.Polyamide resin has special selectivity for flavones, therefore in plant extract industry, can be used as the superior adsorbent of flavonoid compound separation and Extraction.But, the relevant report of not yet useful polyamide resin separation and purification blackberry, blueberry anthocyanidin at present.
Summary of the invention
For solving the deficiencies in the prior art, the object of the present invention is to provide a kind of purification process of blackberry, blueberry anthocyanidin, efficiently purifying can be carried out to blackberry, blueberry anthocyanidin.
In order to realize above-mentioned target, the present invention adopts following technical scheme:
A purification process for blackberry, blueberry anthocyanidin, comprises the following steps: (1) resin pre-treatment: first boiled with ethanol by polyamide resin, then cleans with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: by blackberry, blueberry anthocyanidin crude extract thin up after concentrated volatilization ethanol, after adjust ph, upper polyamide resin column, first washes with water, then uses ethanol elution, concentrates to obtain medicinal extract.
Further, aforementioned purification process, comprises the following steps:
(1) resin pre-treatment: polyamide resin is first boiled 3 ~ 5 hours with 60% ~ 95% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.6 ~ 2.99mgmL after concentrated volatilization ethanol
-1after regulating pH=1 ~ 5 with hydrochloric acid, with the speed upper prop 1 ~ 7BV of 2 ~ 10BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, by pH value be again 40% ~ 100% ethanol of 1 ~ 5 with the speed wash-out 0.5 ~ 5BV of 2 ~ 10BV/h, concentrated to obtain medicinal extract.
Preferably, aforementioned purification process, comprises the following steps:
(1) resin pre-treatment: polyamide resin is first boiled 4 hours with 95% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.75mgmL after concentrated volatilization ethanol
-1, regulate after pH=1 with hydrochloric acid, with the speed upper prop 5BV of 5BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, then with pH value be 80% ethanol of 3 with the speed wash-out 3.5BV of 5BV/h, underpressure distillation concentrates and to obtain medicinal extract.
In aforementioned purification process, the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add the ethanol of water or any concentration, adopt microwave auxiliary extraction, obtain extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
Further, in aforementioned purification process, the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add the ethanol of water or any concentration in the ratio of 5 ~ 25mL/1g blackberry, blueberry slurry, adopt microwave auxiliary extraction, microwave power 100 ~ 700w, microwave treatment time 1 ~ 9min, obtains extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
Preferably, in aforementioned purification process, the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add in the ratio of 25mL/1g blackberry, blueberry slurry the ethanol that concentration is 52%, adopt microwave auxiliary extraction, microwave power 469w, microwave treatment time 4min, obtains extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
In aforementioned purification process, extracting solution is centrifugal is extracting solution under 3500r/min condition centrifugal 3 ~ 8 minutes.
Preferably, in aforementioned purification process, extracting solution is centrifugal is extracting solution under 3500r/min condition centrifugal 5 minutes.
In aforementioned purification process, polyamide resin is 60 ~ 100 orders.
In order to ensure blackberry, blueberry anthocyanidin purification process science of the present invention, rationally, inventors performed corresponding experimental study and screening, just determined technical scheme of the present invention.Specific experiment content is as follows:
One, experiment material, medicine and instrument
Blackberry, blueberry is provided by polar bear Agro-ecology company limited sheep Chinese mugwort blackberry, blueberry plantation, and main agents and device information are in table 1 and table 2, and other reagent used are analytical pure.
Table 1 major experimental reagent information
Table 2 device information
Two, experiment and conclusion
1, purification experiment
1.1 purification experiment processes:
(1) resin pre-treatment:
1. ion exchange resin pre-treatment
Ion exchange resin is soaked in water 12h, then washes with water to non-foam without brown, then carries out wet method dress post.Resin column is rinsed with the flow velocity of 2BV/h with 5%HCl solution, pH=5 is washed till with deionized water again after 2h, then with same flow velocity, resin column is rinsed with 5%NaOH solution, be washed till pH=10 with deionized water again after 2h, then with the flow velocity of 2BV/h, rinse resin column with 5%HCl solution, pH=4 ~ 5 are washed till with deionized water again after 2h, for subsequent use.
2. polyamide resin pre-treatment
Polyamide resin is first boiled 3 ~ 5 hours with 60% ~ 95% ethanol, then cleans to without alcohol taste with distilled water, for subsequent use;
(2) blackberry, blueberry anthocyanidin crude extract preparation:
Blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add the ethanol of water or any concentration in the ratio of 5 ~ 25mL/1g blackberry, blueberry slurry, adopt microwave auxiliary extraction, microwave power 100 ~ 700w, microwave treatment time 1 ~ 9min, obtains extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product;
Described blackberry, blueberry anthocyanidin crude extract solution is that blackberry, blueberry anthocyanidin crude extract thin up after concentrated volatilization ethanol obtains.
(3) purifying of blackberry, blueberry anthocyanidin crude extract:
Blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.6 ~ 2.99mgmL after concentrated volatilization ethanol
-1after regulating pH=1 ~ 5 with hydrochloric acid, with the speed upper prop 1 ~ 7BV of 2 ~ 10BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, by pH value be again 40% ~ 100% ethanol of 1 ~ 5 with the speed wash-out 0.5 ~ 5BV of 2 ~ 10BV/h, concentrated to obtain medicinal extract.
2, blackberry, blueberry anthocyanidin content measures
2.1 blackberry, blueberry anthocyanidin content measuring methods: be with chlorination Cyanidin-3-O-glucose for standard substance are analyzed, measure the anthocyanidin content in blackberry, blueberry anthocyanidin composition.
The drafting of 2.2 typical curves
Accurately weighed 3.34mg anthocyanidin standard substance, dissolve with distilled water, constant volume in 10mL volumetric flask, as mother liquor.Mother liquor being diluted to respectively concentration is 33.4,66.8,100.2,133.6,167,200.4,233.8,267.2 μ g/mL, the anthocyanidin standard substance (blank replaces with 50% ethanol water) getting 1.0mL different concns are respectively placed in tool plug test tube, add the methanol hydrochloride solution of 1.00% of 5.00mL, abundant concussion, 50 DEG C of water-bath 10min, be cooled to room temperature, measure its absorbance at 530nm place (A530).With anthocyanidin standard concentration for X-coordinate (X-axis), absorbancy is ordinate zou (Y-axis), drawing standard curve.Typical curve equation is: Y=0.0033X-0.0092, relation conefficient γ=0.9989, and namely effective constituent concentration range is at 33.4 μ gmL
-1~ 267.2 μ gmL
-1inside there is good linear relationship.Typical curve as shown in Figure 1.
The assay of 2.3 blackberry, blueberry anthocyanidin
Get appropriate blackberry, blueberry anthocyanidin solution (dissolved by purifying gained medicinal extract and obtain), be diluted to suitable concn, carry out chromogenic assay according to 2.2 " drafting of typical curve " method, determine its concentration (c) with typical curve, and then determine the content of blackberry, blueberry anthocyanidin.Specific formula for calculation is such as formula shown in (1):
Note: C: mass concentration (the μ gmL of blackberry, blueberry anthocyanidin
-1); V: liquor capacity (mL); N: extension rate; M: the quality (g) of sample.
3, Static Adsorption and desorb
3.1 Static Adsorption:
The Static Adsorption process of blackberry, blueberry anthocyanidin on resin is carried out in the following manner: all Staticadsorption experiment batch processing modes carry out at 25 DEG C.The blackberry, blueberry anthocyanidin crude extract solution of the sorbent material (pretreated resin) of 5mL and the dilution suitable multiple of 50mL is added in 100mL Erlenmeyer flask respectively, puts on constant temperature oscillator, in 25 DEG C, vibrate under 150 ~ 200r/min condition 24h.After Static Adsorption, surplus solution surveys absorbancy after filtering.Through type (2) calculates the adsorptive power (qe-mgmL of resin when balancing
-1resin).
Note: q
estatic Adsorption ability (mgmL
-1resin); C
0and C
estarting point concentration and the equilibrium concentration (mgmL of anthocyanidin respectively
-1).V
iand V
athe amount of initial sample solution and the amount of initial adsorption agent respectively.
3.2 static desorbs:
Static desorption process carries out in the following manner: after guaranteeing to reach adsorption equilibrium, resin is by washed with de-ionized water, then uses 50mL ethanol (0.1% hydrochloric acid, v/v) wash-out.The solution eluted surveys absorbancy.Through type (3) and formula (4) calculate static desorb balance time static desorption ability and static desorption efficiency (D, %)
Adsorption rate calculates according to formula (9)
Note: q
dstatic desorption ability (mgmL
-1resin), A
oand A
einitial absorbance and the balance absorbancy of blackberry, blueberry anthocyanidin respectively; V
ithe amount of initial sample solution, V
dit is the volume (mL) of elutriant.
Static Adsorption and static desorption experiment is carried out with different meshes polyamide resin.Result as shown in Figure 2.As can be known from Fig. 2,60 ~ 100 orders and 100 ~ 200 order polyamide resins show higher Static Adsorption and static desorption ability relative to the polyamide resin of other order numbers.This is because it has large surface-area.The polyamide resin of the rate of recovery different meshes of blackberry, blueberry anthocyanidin is tested, and result as shown in Figure 3.As can be seen from Figure 3, the rate of recovery of the corresponding anthocyanidin of 60 ~ 100 object polyamide resins is higher than the polyamide resin of other order numbers.Static Adsorption and the static desorption ability of 60 ~ 100 order polyamide resins are respectively 4.41 and 3.29mgmL
-1resin.Because the velocity of separation of 100 ~ 200 order polyamide resins is much smaller than 60 ~ 100 order polyamide resins.Therefore, experiment after this uses 60 ~ 100 order polyamide resins as sorbent material.
4, adsorption isothermal line
The isothermal experiment of equilibrium adsorption of sorbent material carries out in the following manner: add in 100mL Erlenmeyer flask respectively with the blackberry, blueberry anthocyanidin crude extract solution of the sorbent material of 5mL and the different concns of 50mL, be placed on constant temperature oscillator, vibrate 60min respectively under 5 DEG C, 15 DEG C, 25 DEG C conditions.Starting point concentration and equilibrium concentration under differing temps is measured respectively with spectrophotometry.By Lang Gemiaoer and Freundlich model, equilibrium adsorption is described.Lang Gemiaoer thermoisopleth is widely used in monolayer adsorption, and empirical equation is such as formula shown in (5):
Note: K
land q
madsorption equilibrium constant and empirical constant respectively, can by drawing C
e/ q
eand C
elinear graph and corresponding regression coefficient (γ) value obtain.
Freundlich model is widely used in nonideal adsorbed solution, and empirical equation is such as formula shown in (6):
Note: K
ffreundlich constant (adsorptive power); 1/n (adsorption strength) is an empirical constant, can by drawing logq
eand logC
elinear graph and corresponding regression coefficient (γ) value obtain.
The regularity of distribution of blackberry, blueberry anthocyanidin between liquid phase and solid-phase media is determined by the time reached needed for balance in measurement adsorption process, stated by Lang Gemiaoer and Freundlich thermoisopleth.Analyze at 15 DEG C, 25 DEG C, 35 DEG C respectively.The constant of Lang Gemiaoer and Freundlich equation calculates by the slope in Fig. 4 and Fig. 5 and intercept respectively, and result is as shown in table 3.Lang Gemiaoer and Freundlich thermoisopleth are modal models in research desorption balance data.The surface of Freundlich model assumption resin is heterogeneous in itself, is characterized in adsorbing at different energy points.It is described that the absorption behavior of a unimolecular layer and polymolecular layer.Lang Gemiaoer model description one with identical adsorption site as far as possible monolayer adsorption and be there is no mutual absorption behavior each other by the molecule adsorbed.Relation conefficient in table 3 shows that Freundlich model is more suitable for describing equilibrium adsorption.Adsorption strength value (n) of Freundlich model is within the scope of 0<n<10, and this shows that the absorption of blackberry, blueberry anthocyanidin is good.Can see from table, K
fincrease with the reduction of temperature, show that the absorption of blackberry, blueberry anthocyanidin is risen along with temperature and reduces.Thermo-responsive anthocyanidin is degraded at higher temperatures and can be explained these results.In addition, the relation conefficient of Freundlich model is far above Lang Gemiaoer model.Therefore can infer, absorption and the Freundlich equation of blackberry, blueberry anthocyanidin have cognation better, especially under 15 DEG C of conditions.
Table 3
Note: q
max– maximum adsorption ability; γ – regression coefficient; K
l-Lang Gemiaoer constant; K
f-Freundlich constant; N – adsorption strength.
5, adsorption equilibrium and kinetics
The blackberry, blueberry anthocyanidin crude extract solution of the dilution suitable multiple of 5mL sorbent material (pretreated resin) and 50mL is added 100mL Erlenmeyer flask respectively, puts 60min that constant temperature oscillator vibrates.Get 1000 μ L solution every 5min and survey absorbancy, judge that adsorption process reaches the time of balance with this.
By the absorption of pseudo-First order dynamic model and pseudo-second-order equation model analysis blackberry, blueberry anthocyanidin.The relation conefficient (γ value) of the consistence model between experimental data and the predictor of adsorptive power is expressed.Relatively high γ value shows that this model successfully describes the kinetics of adsorption of anthocyanidin.
Pseudo-First order dynamic model, shown in (7):
ln(q
e-q
t)＝lnq
e-K
1t(7)
Formula (7) can be exchanged into
Pseudo-second-order equation model, shown in (8):
Note: K
1and K
2the rate constant of pseudo-First order dynamic model and pseudo-second-order equation model respectively; q
tthe blackberry, blueberry anthocyanidin adsorptive capacity (mgmL when the time is t
-1); q
eit is the adsorptive power when adsorption equilibrium.
Absorption is a physical and chemical processes, relates to a large amount of solute (adsorbate) mass transfer from liquid phase to adsorbent surface.The research of kinetics of adsorption can provide the information about absorption mechanism, may be used for the efficiency assessing adsorption process.The research of kinetics of adsorption describes the solute absorption rate at solid-liquid interface.
In order to assess the suitability of pseudo-First order dynamic model for the absorption of anthocyanidin, depict log (q
e-q
t) with the linear graph of t, as shown in Figure 6.Q
evalue (obtaining from model) and K
1(obtaining from the intercept figure and slope) is as shown in table 4 respectively.T/q
twith the linear graph of t, as shown in Figure 7.Equilibrium adsorption capacities and pseudo-second-order equation constant K
2(obtaining from the intercept figure and slope) is as shown in table 4 respectively.As can be seen from Table 4, the γ (0.9314) of pseudo-First order dynamic model is far below the γ (0.9979) of pseudo-second-order equation model.Can infer thus, pseudo-second-order equation model is more suitable for the absorption describing blackberry, blueberry anthocyanidin.Pseudo-First order dynamic model describes the starting stage of adsorption process.Pseudo-second-order equation model is to prediction and to describe whole adsorption process more suitable.Pseudo-second-order equation model is more suitable for describing the kinetics of adsorption of blackberry, blueberry anthocyanidin on resin than pseudo-First order dynamic model.
Table 4
Note: K
1-pseudo-first order kinetics rate constant; K
2-pseudo-second-order equation rate constant; q
ethe adsorptive power of – kinetic model; γ – regression coefficient.
6, the condition optimizing of absorption and wash-out
After determining adsorption equilibrium and kinetics, adopt single factor experiment to obtain the initial ranges of purifying process parameter, comprise starting point concentration (mgmL
-1), the pH value of adsorption liquid, the pH value of elutriant and alcohol concn (%).
Determine optimum starting point concentration in the following way: the anthocyanidin crude extract solution of the resin sorbent and 20mL different concns of getting 5mL adds in Erlenmeyer flask respectively, put 1h that 15 DEG C of constant temperature oscillators vibrate.Measure not by the absorbancy of blackberry, blueberry anthocyanidin of adsorbing.
Determine the pH value of optimum adsorption liquid in the following way: the adsorption liquid getting the resin sorbent pH value different from 20mL of 5mL adds in Erlenmeyer flask respectively, put 1h that 15 DEG C of constant temperature oscillators vibrate.Measure not by the absorbancy of blackberry, blueberry anthocyanidin of adsorbing.
Determine the pH value of best elutriant in the following way: the elutriant getting the resin sorbent the adsorbing blackberry, blueberry anthocyanidin pH value different from 50mL of 5mL adds in Erlenmeyer flask respectively, put 1h that 15 DEG C of constant temperature oscillators vibrate.Measure the absorbancy of the blackberry, blueberry anthocyanidin eluted.
Determine best alcohol concn in the following way: the ethanol of the resin sorbent adsorbing blackberry, blueberry anthocyanidin and 50mL different concns of getting 5mL adds in Erlenmeyer flask respectively, put 1h that 15 DEG C of constant temperature oscillators vibrate.Measure the absorbancy of the blackberry, blueberry anthocyanidin eluted.
The impact that 6.1 starting point concentrations adsorb blackberry, blueberry anthocyanidin
In order to study the starting point concentration of anthocyanidin to the impact of absorption, adopt at 0.6 ~ 2.99mgmL
-1in scope, the mode of different starting point concentration on the impact of blackberry, blueberry anthocyanidin adsorption rate is stated.Keep pH value=3 of adsorption liquid, pH value=4 of elutriant and alcohol concn are 75%.As shown in Figure 8, adsorption rate with starting point concentration from 2.99 to 0.75mgmL
-1reduction and increase, but reduce starting point concentration further, the increase of adsorption rate is not obvious.Therefore, the preferred starting point concentration of the absorption of blackberry, blueberry anthocyanidin is 0.75mgmL
-1.
The impact that the pH value of 6.2 adsorption liquids is adsorbed blackberry, blueberry anthocyanidin
As shown in Figure 9, adsorption rate reduces with the increase from 1 to 5 of the pH value of adsorption liquid, during pH value=1 of therefore adsorption liquid, is conducive to the absorption of blackberry, blueberry anthocyanidin most.
The pH value of 6.3 elutriants is on the impact of blackberry, blueberry anthocyanidin wash-out
In order to study the pH value of elutriant to the impact of the wash-out of anthocyanidin, adopting and in 1 to 5 scopes, the mode of the impact of desorption efficiency being stated in the pH value of elutriant.Maintenance starting point concentration is 0.75mgmL
-1, adsorption liquid pH value=1 and alcohol concn be 75%.As shown in Figure 10, desorption efficiency (eluting rate) increases along with the increase from 1 to 3 of the pH value of elutriant, peaks, improves the pH value of elutriant further, cause desorption efficiency to reduce when the pH value of elutriant is 3.Therefore the optimum value of the pH value of elutriant is 3.
6.4 alcohol concn are on the impact of blackberry, blueberry anthocyanidin wash-out
As shown in figure 11, along with the increase (from 20% to 100%) of alcohol concn, desorption efficiency (eluting rate) is also in increase, and after alcohol concn reaches 80%, desorption efficiency is on a declining curve again.Therefore the optimal concentration of ethanol is 80%.
7, dynamic adsorption and desorb
7.1 dynamic adsorption and desorption experiment:
Dynamic adsorption and desorption experiment are carried out on chromatography column (20mm × 400mm).The column volume (BV) of sorbent material is 40mL, and loading sorbent material height is 12.7cm.Compare the impact that different flow velocitys (2BV/h, 5BV/h, 10BV/h) adsorbs blackberry, blueberry anthocyanidin.Collect once every 0.5BV, carry out absorbance measurement.Reach absorption saturated after, the sorbent material deionized water (2BV) in post cleans up, and then uses ethanol (pH=3,80%, v/v) wash-out under different in flow rate (2BV/h, 5BV/h, 10BV/h).
7.2 dynamic adsorption and desorption curve
In order to the absorption liquid measure needed for specification and wash-out liquid measure, go to calculation sample leak point of (2BV/h, 5BV/h and 10BV/h) under different in flow rate by experiment.Therefore, by the volume of adsorption liquid and the concentration of solute, obtain the dynamic leakage curve of the polyamide resin of 60 ~ 100 order numbers.
As shown in figure 12.When flow velocity is 5BV/h, absorption property reaches best.When being in low flow velocity, adsorbate molecule and the sorbent material interphase interaction time more, although and high flow rate time required time less, the residence time of adsorbate is short, can impact adsorptive power, reveals stagnation point and also will reach sooner.Usually, outflow concentration reaches 5% of adsorption liquid concentration and is defined as leak point.To sum up, adsorb that liquid measure is 5BV, the top condition of adsorption liquid flow velocity to be 5BV/h be dynamic adsorption.
Drawn by the volume of stripping workshop and solute concentration and obtain dynamic desorption curve, as shown in figure 13.With the 80% ethanol elution anthocyanidin that pH value is 3, keep flow velocity at 2BV/h, 5BV/h and 10BV/h respectively.As can be seen from the figure, be 3.5BV at elutriant consumption, can wash-out anthocyanidin completely when flow velocity is 5BV/h.Therefore, elutriant consumption is 3.5BV, and flow velocity is 5BV/h is best desorption condition.
8, different resins is for the impact of the purifying of blackberry, blueberry anthocyanidin
Selected ion exchange resin, macroporous adsorbent resin and polyamide resin respectively, research different resins, for the impact of the purifying of blackberry, blueberry anthocyanidin, the results are shown in Table 5.Adsorbed by polyamide resin in the blackberry, blueberry anthocyanidin medicinal extract obtained containing 432mgg
-1anthocyanidin, this content is higher than the anthocyanidin purity (160mgg obtained by cationic exchange resin adsorption
-1) and the anthocyanidin purity (176mgg that obtained by macroporous adsorbent resin
-1).Therefore, polyamide resin is optimal selection.
Table 5
Usefulness of the present invention is: the purification process that the invention provides a kind of blackberry, blueberry anthocyanidin, can realize the efficiently purifying of blackberry, blueberry anthocyanidin, and the purity of gained blackberry, blueberry anthocyanidin is high.Purification process of the present invention selects 60 ~ 100 order polyamide resins as sorbent material, and it has large specific surface area and volume particle size, and sorption and desorption ability is high, cost is low, regenerating easily and the characteristic such as preparation method is simple.The present invention is by the research to the sorption and desorption process of blackberry, blueberry anthocyanidin Static and dynamic, optimize purification condition, establish the optimum value of the starting point concentration of blackberry, blueberry anthocyanidin, the pH value of adsorption liquid, the pH value of elutriant and alcohol concn, make the adsorption rate of blackberry, blueberry anthocyanidin and eluting rate all higher.Method of the present invention can realize that blackberry, blueberry anthocyanidin saves time, efficient purifying, is conducive to saving purification time, cost-saving.
Accompanying drawing explanation
Fig. 1 is the typical curve that blackberry, blueberry anthocyanidin content measures;
Fig. 2 is the absorption of blackberry, blueberry anthocyanidin and the elutive power figure that adopt different meshes polyamide resin;
Fig. 3 is the blackberry, blueberry anthocyanidin rate of recovery figure adopting different meshes polyamide resin;
Fig. 4 is C under differing temps
e/ q
eand C
elinear graph;
Fig. 5 is logq under differing temps
eand logC
elinear graph;
Fig. 6 is log (q
e-q
t) with the linear graph of t;
Fig. 7 is t/q
twith the linear graph of t;
Fig. 8 is that blackberry, blueberry anthocyanidin adsorption rate is with starting point concentration variation diagram;
Fig. 9 is that blackberry, blueberry anthocyanidin adsorption rate is with adsorption liquid pH value variation diagram;
Figure 10 is that blackberry, blueberry anthocyanidin eluting rate is with elutriant pH value variation diagram;
Figure 11 is that blackberry, blueberry anthocyanidin eluting rate is with alcohol concn variation diagram;
Figure 12 is dynamic adsorption graphic representation;
Figure 13 is dynamic desorption graphic representation.
Embodiment
Below in conjunction with specific embodiment, the present invention is further introduced.
The purification process of embodiment 1 blackberry, blueberry anthocyanidin, comprises the following steps:
(1) resin pre-treatment: polyamide resin is first boiled 5 hours with 60% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract (by art methods preparation, in this crude extract medicinal extract, the content of blackberry, blueberry anthocyanidin is about 2.8%): blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.6mgmL
-1, regulate after pH=5 with hydrochloric acid, with polyamide resin column 7BV in the speed of 2BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, then with pH value be 40% ethanol of 1 with the speed wash-out 5BV of 2BV/h, concentrate and to obtain medicinal extract.
Adopt this purifying process, the adsorption rate of blackberry, blueberry anthocyanidin is 81%, and eluting rate is 70%, and in purifying gained medicinal extract, the content of blackberry, blueberry anthocyanidin is 62%.
The purification process of embodiment 2 blackberry, blueberry anthocyanidin, comprises the following steps:
(1) resin pre-treatment: 60 ~ 100 order polyamide resins are first boiled 3 hours with 80% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 2.99mgmL after concentrated volatilization ethanol
-1, regulate after pH=3 with hydrochloric acid, with polyamide resin column 1BV in the speed of 10BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, then with pH value be 100% ethanol of 5 with the speed wash-out 0.5BV of 10BV/h, concentrate and to obtain medicinal extract.Adopt this purifying process, the adsorption rate of blackberry, blueberry anthocyanidin is 78%, and eluting rate is 65%, and in purifying gained medicinal extract, the content of blackberry, blueberry anthocyanidin is 60%.
In step (2), blackberry, blueberry anthocyanidin crude extract is according to said method prepared: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add dehydrated alcohol in the ratio of 5mL/1g blackberry, blueberry slurry, adopt microwave auxiliary extraction, microwave power 700w, microwave treatment time 1min, obtains extracting solution; Extracting solution under 3500r/min condition centrifugal 8 minutes, collects supernatant liquor, to obtain final product.After this crude extract is condensed into medicinal extract, the content of blackberry, blueberry anthocyanidin is 3.2%.
The purification process of embodiment 3 blackberry, blueberry anthocyanidin, comprises the following steps:
(1) resin pre-treatment: 80 order polyamide resins are first boiled 4 hours with 95% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.75mgmL after concentrated volatilization ethanol
-1, regulate after pH=1 with hydrochloric acid, with polyamide resin column 5BV in the speed of 5BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, then with pH value be 80% ethanol of 3 with the speed wash-out 3.5BV of 5BV/h, underpressure distillation concentrates and to obtain medicinal extract.Adopt this purifying process, the adsorption rate of blackberry, blueberry anthocyanidin is 85%, and eluting rate is 78%, and in purifying gained medicinal extract, the content of blackberry, blueberry anthocyanidin is 66%.
In step (2), blackberry, blueberry anthocyanidin crude extract is according to said method prepared: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add in the ratio of 25mL/1g blackberry, blueberry slurry the ethanol that concentration is 52%, adopt microwave auxiliary extraction, microwave power 469w, microwave treatment time 4min, obtains extracting solution; Extracting solution under 3500r/min condition centrifugal 5 minutes, collects supernatant liquor, to obtain final product.After this crude extract is condensed into medicinal extract, the content of blackberry, blueberry anthocyanidin is 5.2%.
The purification process of embodiment 4 blackberry, blueberry anthocyanidin, comprises the following steps:
(1) resin pre-treatment: 60 ~ 70 order polyamide resins are first boiled 4.5 hours with 75% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 1.5mgmL after concentrated volatilization ethanol
-1, regulate after pH=2 with hydrochloric acid, with polyamide resin column 4BV in the speed of 8BV/h, wash 3BV (be washed till and almost do not have sugar) subsequently with water, then with pH value be 90% ethanol of 4 with the speed wash-out 3BV of 4BV/h, underpressure distillation concentrates and to obtain medicinal extract.Adopt this purifying process, the adsorption rate of blackberry, blueberry anthocyanidin is 80%, and eluting rate is 67%, and in purifying gained medicinal extract, the content of blackberry, blueberry anthocyanidin is 61%.
In step (2), blackberry, blueberry anthocyanidin crude extract is according to said method prepared: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add water in the ratio of 15mL/1g blackberry, blueberry slurry, adopt microwave auxiliary extraction, microwave power 100w, microwave treatment time 9min, obtains extracting solution; Extracting solution under 3500r/min condition centrifugal 3 minutes, collects supernatant liquor, to obtain final product.After this crude extract is condensed into medicinal extract, the content of blackberry, blueberry anthocyanidin is 4.1%.
Claims (9)
1. a purification process for blackberry, blueberry anthocyanidin, is characterized in that: comprise the following steps:
(1) resin pre-treatment: polyamide resin is first boiled with ethanol, then clean with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: by blackberry, blueberry anthocyanidin crude extract thin up after concentrated volatilization ethanol, after adjust ph, upper polyamide resin column, first washes with water, then uses ethanol elution, concentrates to obtain medicinal extract.
2. the purification process of blackberry, blueberry anthocyanidin according to claim 1, is characterized in that: comprise the following steps:
(1) resin pre-treatment: polyamide resin is first boiled 3 ~ 5 hours with 60% ~ 95% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.6 ~ 2.99mgmL after concentrated volatilization ethanol
-1, after regulating pH=1 ~ 5 with hydrochloric acid, with the speed upper prop 1 ~ 7BV of 2 ~ 10BV/h, wash 3BV with water subsequently, then by pH value be 40% ~ 100% ethanol of 1 ~ 5 with the speed wash-out 0.5 ~ 5BV of 2 ~ 10BV/h, concentratedly to obtain medicinal extract.
3. the purification process of blackberry, blueberry anthocyanidin according to claim 2, is characterized in that: comprise the following steps:
(1) resin pre-treatment: polyamide resin is first boiled 4 hours with 95% ethanol, then clean to without alcohol taste with distilled water, for subsequent use;
(2) purifying of blackberry, blueberry anthocyanidin crude extract: blackberry, blueberry anthocyanidin crude extract is diluted with water to 0.75mgmL after concentrated volatilization ethanol
-1, regulate after pH=1 with hydrochloric acid, with the speed upper prop 5BV of 5BV/h, wash 3BV with water subsequently, then with pH value be 80% ethanol of 3 with the speed wash-out 3.5BV of 5BV/h, underpressure distillation concentrates and to obtain medicinal extract.
4. the purification process of the blackberry, blueberry anthocyanidin according to any one of claims 1 to 3, is characterized in that: the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add the ethanol of water or any concentration, adopt microwave auxiliary extraction, obtain extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
5. the purification process of blackberry, blueberry anthocyanidin according to claim 4, is characterized in that: the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add the ethanol of water or any concentration in the ratio of 5 ~ 25mL/1g blackberry, blueberry slurry, adopt microwave auxiliary extraction, microwave power 100 ~ 700w, microwave treatment time 1 ~ 9min, obtains extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
6. the purification process of blackberry, blueberry anthocyanidin according to claim 5, is characterized in that: the preparation method of described blackberry, blueberry anthocyanidin crude extract is: blackberry, blueberry is carried out homogenate, obtains blackberry, blueberry slurry for subsequent use; Add in the ratio of 25mL/1g blackberry, blueberry slurry the ethanol that concentration is 52%, adopt microwave auxiliary extraction, microwave power 469w, microwave treatment time 4min, obtains extracting solution; Extracting solution is centrifugal, collects supernatant liquor, to obtain final product.
7. the purification process of blackberry, blueberry anthocyanidin according to claim 4, is characterized in that: described extracting solution is centrifugal is extracting solution under 3500r/min condition centrifugal 3 ~ 8 minutes.
8. the purification process of blackberry, blueberry anthocyanidin according to claim 7, is characterized in that: described extracting solution is centrifugal is extracting solution under 3500r/min condition centrifugal 5 minutes.
9. the purification process of blackberry, blueberry anthocyanidin according to claims 1 to 3, is characterized in that: described polyamide resin is 60 ~ 100 orders.
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| CN107114786A (en) * | 2017-05-08 | 2017-09-01 | 贵州师范大学 | A kind of method for lifting blackberry, blueberry anthocyanidin stability |
| CN107744673A (en) * | 2017-08-18 | 2018-03-02 | 佛山科学技术学院 | A kind of cockscomb callus anthocyanidin extract solution |
| CN115537036A (en) * | 2022-10-27 | 2022-12-30 | 浙江中医药大学 | Process for purifying carthamus tinctorius red pigment by macroporous resin |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107114786A (en) * | 2017-05-08 | 2017-09-01 | 贵州师范大学 | A kind of method for lifting blackberry, blueberry anthocyanidin stability |
| CN107114786B (en) * | 2017-05-08 | 2020-11-24 | 贵州师范大学 | Method for improving stability of blackberry anthocyanin |
| CN107744673A (en) * | 2017-08-18 | 2018-03-02 | 佛山科学技术学院 | A kind of cockscomb callus anthocyanidin extract solution |
| CN115537036A (en) * | 2022-10-27 | 2022-12-30 | 浙江中医药大学 | Process for purifying carthamus tinctorius red pigment by macroporous resin |
| CN115537036B (en) * | 2022-10-27 | 2024-05-10 | 浙江中医药大学 | Process for purifying safflower red pigment by macroporous resin |
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