CN103169789A - Method for extracting total flavonoid in silverweed cinquefoil root - Google Patents
Method for extracting total flavonoid in silverweed cinquefoil root Download PDFInfo
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- CN103169789A CN103169789A CN2012105959928A CN201210595992A CN103169789A CN 103169789 A CN103169789 A CN 103169789A CN 2012105959928 A CN2012105959928 A CN 2012105959928A CN 201210595992 A CN201210595992 A CN 201210595992A CN 103169789 A CN103169789 A CN 103169789A
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Abstract
The invention belongs to the technical field of extraction of total flavonoid in silverweed cinquefoil root and provides a method for extracting total flavonoid in silverweed cinquefoil root. The method comprises the following steps of: by using water as an extracting agent, adding a 0.35% surface active agent triethanolamine solution into the water under a pretreatment conditions that the material to liquid ratio (g/ml) is 1:60 and the solution is preboiled for 10 minutes, treating the solution by microwave for 2 minutes, wherein the power of the microwave is 373 W, and the extraction rate of the total flavonoid in the silverweed cinquefoil root is 1.326% under the technical condition. The method disclosed by the invention is simple in technical process, has low requirements on production equipment and technique, is low in extraction cost, achieves high extraction rate of total flavonoid, and has high promotion and application values.
Description
Technical field
The invention belongs to Radix potentillae anserinae total flavones extractive technique field, relate in particular to a kind of method of extracting total flavones in Radix potentillae anserinae.
Background technology
Radix potentillae anserinae mainly originates in the extremely frigid zones of the height above sea level 1700～4300m such as Qinghai, Tibet, Gannan, the formal name used at school Potentilla anserina L., Rosaceae belongs to the perennial herbaceous plant that crawls, it not only contains the nutrients such as aminoacid, electrolytes and minerals, and contains the bioactive substances such as flavonoid.Flavonoid substances has allaying tiredness, protects blood vessel, prevents arteriosclerosis, expands blood capillary, dredges microcirculation, the anti-ageing function of waiting for a long time.In recent years, people compare deep research to anti-oxidation function and the mechanism of flavone compound, confirm that flavone compound is a kind of very strong reactive oxygen free radical scavenger, so the research of flavone compound has caused people's attention day by day.
The method that tradition is extracted flavone mainly contains: solvent extraction method, alkaline extraction acid precipitation method etc., Microwave Assisted Extraction Technique, it is a kind of novel extraction technology of rising the eighties in 20th century, have efficient fast, selectivity is good, solvent load is few, the response rate is high, high repeatability and other advantages, is the study hotspot of natural product extraction both at home and abroad at present.Even surfactant is a class also remarkable material of reduction table (boundary) surface tension when concentration is very low.There is now researcher to utilize solubilising, the wetting action of surfactant, carried out the collaborative research of extracting natural product of surfactant, greatly improved extraction ratio.The research report that extracts Flavonoid substances in Radix potentillae anserinae about microwave extracting is few, and particularly the method for the collaborative microwave extracting of surfactant has no bibliographical information.This paper is intended to study the process conditions of total flavones in the collaborative microwave extracting Radix potentillae anserinae of surfactant, in order to provide theoretical foundation for the utilization of Radix potentillae anserinae.
Summary of the invention
The invention provides a kind of method of extracting total flavones in Radix potentillae anserinae, be intended to solve the method for total flavones in the extraction Radix potentillae anserinae that prior art provides, complex technical process, extraction cost are higher, the problem that the extraction ratio of total flavones is lower.
The object of the present invention is to provide a kind of method of extracting total flavones in Radix potentillae anserinae, the method comprising the steps of:
Step 2 takes appropriate Radix potentillae anserinae powder, adds the aqueous surfactant solution of certain mass mark, precooks 10 minutes in 80 ℃ of water-baths;
Step 3 was placed 30 minutes in 80 ℃ of water-baths after microwave treatment, and extracting solution is sucking filtration while hot, collected liquid, and distilling under reduced pressure is concentrated, and carried out standardize solution and process.
Further, the optimum process condition that the method is extracted total flavones in Radix potentillae anserinae is: take water as extractant, under pretreatment condition, adding mass fraction is 0.35% triethanolamine solution, microwave treatment time 2 minutes, 373 watts of microwave powers.
Further, pretreatment condition is: solid-liquid ratio (g/ml) 1: 60, precooked 10 minutes for 80 ℃.
Further, Radix potentillae anserinae total flavones extraction ratio is 1.326% under these process conditions.
The method of total flavones in extraction Radix potentillae anserinae provided by the invention, the method is take water as extractant, under 1: 60,80 ℃ pretreatment conditions of precooking 10 minutes of solid-liquid ratio (g/ml), adding mass fraction is 0.35% surfactant triethanolamine solution, microwave treatment time 2 minutes, 373 watts of microwave powers, Radix potentillae anserinae total flavones extraction ratio is 1.326% under these process conditions, technical process is simple, lower to the requirement of production equipment and technology, extraction cost is lower, the extraction ratio of total flavones is higher, has stronger propagation and employment and is worth.
Description of drawings
Fig. 1 is the realization flow figure of the method for total flavones in the extraction Radix potentillae anserinae that provides of the embodiment of the present invention;
Fig. 2 is the control curve of the rutin standard substance that provide of the embodiment of the present invention;
Fig. 3 is that the different surfaces activating agent that provides of the embodiment of the present invention is on the flow chart of extraction ratio impact;
Fig. 4 is that the dosage of surfactant that provides of the embodiment of the present invention is on the curve synoptic diagram of total flavones extraction ratio impact in Radix potentillae anserinae;
Fig. 5 is that the microwave power that provides of the embodiment of the present invention is on the curve synoptic diagram of total flavones extraction ratio impact in Radix potentillae anserinae;
Fig. 6 is that microwave time of providing of the embodiment of the present invention is on the curve synoptic diagram of total flavones extraction ratio impact in Radix potentillae anserinae;
Fig. 7 is that the solid-liquid ratio that provides of the embodiment of the present invention is on the curve synoptic diagram of total flavones extraction ratio impact in Radix potentillae anserinae.
The specific embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is described in further detail.Should be appreciated that specific embodiment described herein only in order to explaining the present invention, and be not used in and limit invention.
Fig. 1 shows the realization flow of the method for total flavones in the extraction Radix potentillae anserinae that the embodiment of the present invention provides.
The method comprising the steps of:
Step S101 gets appropriate Radix potentillae anserinae, carries out pulverization process after drying;
Step S102 takes appropriate Radix potentillae anserinae powder, adds the aqueous surfactant solution of certain mass mark, precooks 10 minutes in 80 ℃ of water-baths;
Step S103 placed 30 minutes in 80 ℃ of water-baths after microwave treatment, and extracting solution is sucking filtration while hot, collected liquid, and distilling under reduced pressure is concentrated, and carried out standardize solution and process.
In embodiments of the present invention, the optimum process condition that the method is extracted total flavones in Radix potentillae anserinae is: take water as extractant, under pretreatment condition, adding mass fraction is 0.35% triethanolamine solution, microwave treatment time 2 minutes, 373 watts of microwave powers.
In embodiments of the present invention, pretreatment condition is: solid-liquid ratio (g/ml) 1: 60, precooked 10 minutes for 80 ℃.
In embodiments of the present invention, Radix potentillae anserinae total flavones extraction ratio is 1.326% under these process conditions.
Below in conjunction with drawings and the specific embodiments, application principle of the present invention is further described.
Material and reagent:
Radix potentillae anserinae, rutin standard substance (purity 〉=98%), dehydrated alcohol, sodium nitrite, aluminum nitrate, sodium hydroxide, triethanolamine, dodecylbenzene sodium sulfonate, sodium lauryl sulphate, cetyl trimethyl ammonium bromide are analytical pure.
Instrument:
722 type visible spectrophotometers, the desk-top refrigerated centrifuge of L500, rotary evaporator, the multiplex vacuum pump of SHB-III circulating water type, microwave oven, electric-heated thermostatic water bath, electronic balance, pipettor, electric heating constant-temperature blowing drying box, triangular flask.
Technological process:
Radix potentillae anserinae → drying → pulverizing → pretreatment → microwave extraction → water-bath → filtration → concentrated → standardize solution → mensuration
Get appropriate Radix potentillae anserinae, dry 2h in the baking oven of 60 ℃ crosses 40 mesh sieves after pulverizing, accurately taking the 1.5g Radix potentillae anserinae powder is put in triangular flask, the aqueous surfactant solution that adds the certain mass mark was precooked 10 minutes in 80 ℃ of water-baths, placed 30 minutes in 80 ℃ of water-baths after microwave treatment.Extracting solution is sucking filtration while hot, collects liquid, and distilling under reduced pressure is concentrated, is settled to 25ml, with the absorbance of spectrophotometric determination filtrate, calculates extraction ratio.
The drafting of standard curve:
Accurately take the 0.1326g rutoside standard sample, in the brown volumetric flask of 500mL, stand-by with 30% dissolve with ethanol and standardize solution.Pipette respectively above-mentioned rutin titer 0.0,1.0,2.0,4.0,6.0,8.0mL in 6 50mL volumetric flasks, respectively add 1.0mL 50g/L NaNO
2The solution mixing adds 1.0mL 100g/L Al (NO after placement 6min
3)
3The solution mixing, place 6min, add again 10.0mL 40g/L NaOH solution, be settled to scale with 30% ethanol at last, mixing, after standing 15min, in the 500nm wavelength absorbance A of place, blank reagent is made reference, take absorbance A as vertical coordinate, rutin mass concentration C (mg/ml) is abscissa drawing standard curve.Fig. 2 is the standard curve of drawing, and the regression equation of light absorption value A and rutin concentration C is: A=10.1464C-0.0006, R
2=0.9997.
The mensuration of Radix potentillae anserinae total flavones extraction ratio:
Accurately draw sample liquid 1ml in 25ml tool plug test tube, add 1.0mL 50g/L NaNO
2The solution mixing adds 1.0mL 100g/L Al (NO after placement 6min
3)
3The solution mixing is placed 6min, then is added 10.0mL40g/L NaOH solution, is settled to scale with 30% ethanol at last, then places 15min, does simultaneously reagent blank and is contrast, measures light absorption value also by regression equation calculation in the 500nm wavelength.In Radix potentillae anserinae, the total flavones extraction ratio calculates by following formula:
Total flavones extraction ratio (%)=c * v
1* n/ (w * 1000) * 100%
In formula: the flavone concentration that c-obtains according to regression equation calculation, mg/ml;
v
1-be the sample liquid cumulative volume, ml;
N-is extension rate;
W-is the Radix potentillae anserinae quality, g.
Single factor experiment:
The impact of kinds of surfactants on extraction ratio
Accurately take the Radix potentillae anserinae powder 1.5g of equivalent, triethanolamine take mass fraction as 0.40%, dodecylbenzene sodium sulfonate, sodium lauryl sulphate, cetyl trimethyl ammonium bromide aqueous solution are as extractant, after pretreatment, be 1: 30 (g/ml) at solid-liquid ratio, microwave power is 480w, extraction time is to carry out microwave extraction under the condition of 3min, and the test triplicate is investigated kinds of surfactants to the impact of extraction ratio.
As seen from Figure 3, under similarity condition, test in several surfactants used, extraction ratio order from big to small is followed successively by: triethanolamine, sodium lauryl sulphate, dodecylbenzene sodium sulfonate, cetyl trimethyl ammonium bromide.The triethanolamine extraction effect is best, therefore determine that triethanolamine uses surfactant for experiment.
The impact of dosage of surfactant on extraction ratio
Accurately take the Radix potentillae anserinae powder 1.5g of equivalent, be respectively 0.05%, 0.2%, 0.35%, 0.5%, 0.65% triethanolamine aqueous solution as extractant take mass fraction, after pretreatment, be 1: 30 (g/ml) at solid-liquid ratio, microwave power is 480w, extraction time is to carry out microwave extraction under the condition of 3min, and the test triplicate is investigated dosage of surfactant to the impact of extraction ratio.
As seen from Figure 4, along with the increase of triethanolamine solution mass fraction, extraction ratio constantly increases, and when liquid quality fraction reached 0.2%, extraction rate reached was to maximum, and continuing increases liquid quality fraction, and extraction ratio descends on the contrary.Hence one can see that, the too high or too low optimum extraction effect that all do not reach of triethanolamine solution mass fraction, and the concentration of surfactant is larger, more easily forms micellar structure, thereby increases the water solublity of effective ingredient, and the flavone stripping is increased.Yet along with the further increase of surfactant mass fraction, extraction ratio is on a declining curve, and this may be because surfactant not only has solubilization to flavone, simultaneously other impurity is also had solubilization, causes purity drop.Historical facts or anecdotes is tested and selected mass fraction is that 0.2% triethanolamine solution is as the best in quality mark that extracts total flavones in Radix potentillae anserinae.
The impact of microwave power on extraction ratio:
Accurately take the Radix potentillae anserinae powder 1.5g of equivalent, triethanolamine aqueous solution take mass fraction as 0.2% is as extractant, after pretreatment, be 1: 30 (g/ml) at solid-liquid ratio, microwave power is respectively 160w, 267w, 373w, 480w, 587w, extraction time is to carry out microwave extraction under the condition of 3min, and the test triplicate is investigated microwave power to the impact of extraction ratio.
As seen from Figure 5, extraction ratio begins to raise along with the increase of microwave power, and when microwave power is 373W, extraction ratio is maximum.This may be because microwave power raises, temperature also increases, make cell rupture, be beneficial to the stripping of Flavonoid substances, when microwave power continues to increase, cause temperature too high, may cause that flavone decomposes, extraction ratio descends, and therefore selecting microwave power is that 373W is as the best microwave power that extracts total flavones in Radix potentillae anserinae.
The impact of microwave time on extraction ratio:
Accurately take the Radix potentillae anserinae powder 1.5g of equivalent, triethanolamine aqueous solution take mass fraction as 0.2% is as extractant, after pretreatment, be 1: 30 (g/ml) at solid-liquid ratio, microwave power is 373W, extraction time is respectively under the condition of 1min, 3min, 5min, 7min, 9min, carries out microwave extraction.The test triplicate is investigated the microwave time to the impact of extraction ratio.
As seen from Figure 6, extraction ratio increases with the prolongation of extraction time, and when extraction time was 3min, extraction ratio was maximum.When extraction time continues to increase, extraction ratio descends on the contrary, reason may be that microwave extraction can directly pass to cell interior to energy, Fast Heating, be conducive to the flavone stripping, but it is stable that long-time heating is unfavorable for flavone, causes extraction ratio to descend, and therefore selecting the microwave time is that 3min is as the best microwave extraction time of extracting total flavones in Radix potentillae anserinae.
The impact of solid-liquid ratio on extraction ratio:
Accurately take the Radix potentillae anserinae powder 1.5g of equivalent, triethanolamine aqueous solution take mass fraction as 0.2% is as extractant, after pretreatment, be respectively 1: 30,1: 40,1: 50,1: 60,1: 70 (g/ml) at solid-liquid ratio, microwave power is 373W, and extraction time is to carry out microwave extraction under the condition of 3min.The test triplicate is investigated solid-liquid ratio to the impact of extraction ratio.
As seen from Figure 7, extraction ratio is maximum when liquid ratio is 1: 50 (g/ml), after this present the trend that reduces along with the increase of solid-liquid ratio, may be due to most strippings of the flavone in Radix potentillae anserinae, increase solvent volume again and may cause due to the increase of some other impurity and pigment the reduction of total flavones extraction ratio, therefore select solid-liquid ratio to be: 1: 50 (g/ml) is as the best solid-liquid ratio that extracts total flavones in Radix potentillae anserinae.
Orthogonal test:
On the basis of single factor experiment, design L
9(3
4) orthogonal test research determines the optimum process condition of the collaborative microwave extraction Radix potentillae anserinae flavone of surfactant.
Table 1 orthogonal test factor level table
Table 2 L
9(3
4) orthogonal experiments
As shown in Table 2, the order that each factor affects Radix potentillae anserinae flavone extraction ratio is A＞B＞C＞D, and namely the factor that has the greatest impact of Radix potentillae anserinae total flavones extraction ratio is the mass fraction of surfactant, is then microwave power, microwave time, solid-liquid ratio successively; Best extraction conditions is A
3B
2C
1D
3, namely surfactant triethanolamine mass fraction is 0.35%, microwave power 373W, microwave extraction time 2min, solid-liquid ratio are 1: 60 (g/ml).
Adopting the optimum process condition of surfactant assisted extraction Radix potentillae anserinae total flavones is that the surfactant mass fraction is 0.35%, microwave power 373W, microwave extraction time 2min, solid-liquid ratio is under the condition of 1: 60 (g/ml), and Radix potentillae anserinae total flavones extraction ratio is 1.326%.Result shows, utilizes surfactant solution to extract total flavones in Radix potentillae anserinae, can reduce extraction cost, and the method technique is simple, for the novel technology for extracting research of Radix potentillae anserinae total flavones lays the foundation.
The method of total flavones in the extraction Radix potentillae anserinae that the embodiment of the present invention provides, the method is take water as extractant, under 1: 60,80 ℃ pretreatment conditions of precooking 10 minutes of solid-liquid ratio (g/ml), adding mass fraction is 0.35% surfactant triethanolamine solution, microwave treatment time 2 minutes, 373 watts of microwave powers, Radix potentillae anserinae total flavones extraction ratio is 1.326% under these process conditions, technical process is simple, lower to the requirement of production equipment and technology, extraction cost is lower, the extraction ratio of total flavones is higher, has stronger propagation and employment and is worth.
The above is only preferred embodiment of the present invention, not in order to limiting the present invention, all any modifications of doing within the spirit and principles in the present invention, is equal to and replaces and improvement etc., within all should being included in protection scope of the present invention.
Claims (4)
1. a method of extracting total flavones in Radix potentillae anserinae, is characterized in that, the method comprises the following steps:
Step 1 is got appropriate Radix potentillae anserinae, and 60 ℃ of dry 2h in baking oven cross 40 mesh sieves after pulverizing;
Step 2 takes appropriate Radix potentillae anserinae powder, adds the aqueous surfactant solution of certain mass mark, precooks 10 minutes in 80 ℃ of water-baths;
Step 3 was placed 30 minutes in the water-bath of 80 ℃ after microwave treatment, and extracting solution is sucking filtration while hot, collected liquid, and distilling under reduced pressure is concentrated, and carried out standardize solution and process.
2. the method for claim 1, it is characterized in that, the optimum process condition that the method is extracted total flavones in Radix potentillae anserinae is: take water as extractant, under pretreatment condition, adding mass fraction is 0.35% triethanolamine solution, microwave treatment time 2 minutes, 373 watts of microwave powers.
3. method as claimed in claim 2, is characterized in that, pretreatment condition is: solid-liquid ratio (g/ml) 1: 60, precooked 10 minutes for 80 ℃.
4. method as claimed in claim 2, is characterized in that, Radix potentillae anserinae total flavones extraction ratio is 1.326% under these process conditions.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5928646A (en) * | 1996-05-30 | 1999-07-27 | Berkem | Process for extracting catechin polyphenols from potentillas, extract obtained and its use |
| CN101358222A (en) * | 2008-09-01 | 2009-02-04 | 西北师范大学 | Method for microwave auxiliarily extracting antiradiation polysaccharide using potentilla anserine as feedstock |
| CN101912445A (en) * | 2010-08-26 | 2010-12-15 | 西北农林科技大学 | Method for extracting general flavonoids from toona sinensis leaves under assistance of surfactant |
-
2012
- 2012-12-18 CN CN2012105959928A patent/CN103169789A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5928646A (en) * | 1996-05-30 | 1999-07-27 | Berkem | Process for extracting catechin polyphenols from potentillas, extract obtained and its use |
| CN101358222A (en) * | 2008-09-01 | 2009-02-04 | 西北师范大学 | Method for microwave auxiliarily extracting antiradiation polysaccharide using potentilla anserine as feedstock |
| CN101912445A (en) * | 2010-08-26 | 2010-12-15 | 西北农林科技大学 | Method for extracting general flavonoids from toona sinensis leaves under assistance of surfactant |
Non-Patent Citations (2)
| Title |
|---|
| 李军乔: "采收季节对鹅绒委陵菜(Potentilla anserina L.)块根总黄酮含量的影响", 《青海师范大学学报(自然科学版)》, no. 4, 31 December 2004 (2004-12-31) * |
| 韩志萍: "青海蕨麻中类黄酮的提取及其抗氧化性研究", 《食品研究与开发》, vol. 27, no. 1, 28 February 2006 (2006-02-28) * |
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Application publication date: 20130626 |