CN108732117A - A kind of extracting method of the Radix Glehniae saccharide compound optimized using response phase method combination genetic algorithm and Direct search algorithm - Google Patents
A kind of extracting method of the Radix Glehniae saccharide compound optimized using response phase method combination genetic algorithm and Direct search algorithm Download PDFInfo
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Abstract
The invention belongs to compound extractive technique fields, and in particular to a kind of extracting method of the Radix Glehniae saccharide compound optimized using response phase method combination genetic algorithm and Direct search algorithm.This method is realized by following steps：（1）The preparation of test solution；（2）The making of standard curve；（3）Experimental design and statistical analysis：1. single factor experiment 2. BBD- response phase methods optimization analysis；（4）Genetic algorithm searching is analyzed；（5）Direct search method searching analysis.After the present invention first uses BBD- response surface optimization methods founding mathematical models, acquisition object function, the accurate optimal value of each factor is searched for using genetic algorithm and Direct search algorithm, three kinds of methods are used in combination, to obtain optimum optimization as a result, overcoming the limitation present in existing three kinds of methods.The extracting method optimized through the invention can realize and be extracted to the greatest extent to Radix Glehniae saccharide compound have important research significance.
Description
Technical field
The invention belongs to compound extractive technique fields, and in particular to a kind of using response phase method combination genetic algorithm and straight
Connect the extracting method of the Radix Glehniae saccharide compound of searching algorithm optimization.
Background technology
Radix Glehniae (Radix Glehniae) is samphire glehnia littoralisGlehnia littoralisFr. schmidt
The dry root of ex Miq.The moral character is sweet, and mildly bitter flavor is slightly cold, and plays the role of nourishiing yin to clear away the lung-heat, reinforcing stomach reg fluid, dry for treatment lung heat
Cough, pyreticosis Tianjin wound, the illnesss such as phlegm blood, thirsty of coughing of chattering, are one of our province genunie medicinal materials.The main chemical compositions of Radix Glehniae are sugar
Class, Coumarins, phosphatide, alkaloid, amino acid, volatile oil, trace element etc..Wherein, carbohydrate content content highest.In addition,
Some researches show that Radix Glehniae saccharide compounds can enhance humoral immunity and cellular immune function, has immunocompetence.Therefore, north is husky
The main component of ginseng（Content is high）With important immune active ingredient --- saccharide compound has important researching value, to northern husky
The optimizing research that ginseng saccharide compound extracts condition necessitates.
Document《The technical study of Star point design-response phase method optimization hot water extraction Glehnia Littoralis Polysaccharide》And《Response phase method is excellent
Change the Study on extraction of carbohydrate in Radix Glehniae》It is excellent to provide a kind of technique carrying out saccharide compound extraction using response phase method
Change method, still, both of which method all only simply use response phase method and carry out process optimization, the fluctuation pair of each factor in method
The influence of effect value is indefinite, in actual application, in order to reach higher effect value, needs to carry out each factor accurate
Control, increase the difficulty of technology controlling and process, presently, there are method in, for problems, there are no preferred solutions.
Study a kind of more optimal method of extraction process for the extraction of medicinal material Radix Glehniae saccharide compound have important value and
Meaning.
Invention content
For problems of the prior art, the present invention provides a kind of using response phase method combination genetic algorithm and straight
Connect the extracting method of the Radix Glehniae saccharide compound of searching algorithm optimization.
Used technical solution is the present invention to achieve the goals above：
The present invention provides a kind of Radix Glehniae carbohydrates optimized using response phase method combination genetic algorithm and Direct search algorithm
The extracting method for closing object, includes the following steps：
（1）The preparation of test solution：50-200 mesh coastal glehnia root 5g are weighed, are carried with ratio of water to material 10-60mL/g hot water returns
It takes 1-3 times, extraction time 1-6h, extracting solution filter after merging, centrifuges, Aspirate supernatant is settled to 500mL volumetric flasks, therefrom
Precision measures 4mL and is settled to 250mL volumetric flasks；It is therefrom accurate after mixing well to draw in the tool plug test tube that 1mL enters dried and clean,
Parallel 3 test tubes continue Anthrone Sulphuric acid reaction, then measure the absorbance of test solution, each sample at 590 nm
Product measure twice, with the average value of absorbanceCalculate recovery rate result；
（2）The making of standard curve：105 DEG C of dryings are weighed to the DEXTROSE ANHYDROUS 0.5g of constant weight, with pure water dilution and constant volume
To 100mL volumetric flasks, therefrom draws 1mL and be settled to 50mL volumetric flasks with pure water again, obtain the grape of a concentration of 0.1mg/mL
Standard for Sugars solution；Absorption 0.2,0.4,0.6,0.8,1.0,1.2mL glucose standards solutions respectively, every group of parallel 3 test tube, often
It is 1.2mL that branch test tube adds pure water to total liquid volume respectively, while 1.2mL pure water being taken to make blank control, is separately added into
4mL anthrone reagents, cooling in immersion ice-water bath rapidly, after shake well, each test tube immerses in boiling water bath simultaneously, and nozzle capping is opened
Beginning reaction, reacts 10 minutes, and shaking is uniform, cooling, is placed at room temperature for after ten minutes, obtains standard sample, and detect wave in 590nm
Under length, the absorbance of each standard sample is measured;Each sample measures twice, with the average value of absorbanceFor ordinate, with Portugal
The concentration C of grape sugar（mg/mL）For abscissa, the calibration curve equation of glucose is obtained；
（3）Experimental design and statistical analysis：1. single factor experiment：It is with medicinal material granularity, extraction time, ratio of water to material and extraction time
Parameter carries out single factor experiment, measures the saccharide compound content in Radix glehniae extract using Anthrone-sulfuricacid method, it is husky to calculate north
Join saccharide compound recovery rate（%）, calculation formula is：Recovery rate（%）The volume * of the mass concentration * solution of=saccharide compound is dilute
Release the quality * 100 of multiple/Radix Glehniae medicinal material；2. the optimization analysis of BBD- response phase methods：On the basis of single factor experiment, use
Software Design-Expert 8.0.5b, using the recovery rate of Radix Glehniae saccharide compound as response（Y）, during selection and withdrawal
The significant medicinal material granularity of influential effect（A）, extraction time（B）And ratio of water to material（C）Three factors are independent variable, establish polynary secondary returning
Return equation：Y=-69.04440+0.88849A+14.07365B+0.49493C-0.040163AB-0.0000804375AC+
0.055938BC-0.00232195A2-1.47066B2-0.00669713C2（R 2 =0.8915,P=0.0115）；
Software analysis is carried out, response surface design figure and contour map are obtained；
（4）Genetic algorithm searching is analyzed：Using BBD binomial fittings equation as object function, using software Matlab 2014b, adopt
With the optimum extraction condition of Genetic algorithm searching Radix Glehniae saccharide compound, search process uses genetic algorithm tool GUI, setting
Parameter, random search 10 times, verification analysis；
（5）Direct search method searching analysis：Using binomial fitting equation as object function, using software Matlab 2014b, adopt
With Direct search algorithm search for Radix Glehniae saccharide compound optimum extraction condition, search process use pattern research tool GUI,
Arrange parameter, random search 10 times, verification analysis.
Further, step（4）In, the search process of the genetic algorithm uses genetic algorithm tool GUI, arrange parameter
As follows：
。
Further, step（5）In, the search process use pattern research tool GUI of the direct search method, setting ginseng
Number is as follows：
。
After the present invention is by optimization, the extracting method of optimization is：Radix Glehniae pharmaceutical decocting piece is taken, is crushed as 150 mesh of granularity
Powder, precision weighs Radix Glehniae medicinal powder, and hot water return extraction, 49: 1 mL/g of ratio of water to material is centrifuged after extracting solution filtration
Supernatant is retained in separation, and sediment fraction adds water to repeat last fetched process, and amount of water is identical, and each extraction time is 3.2h,
It extracts 2 times altogether.
Further, the condition of the centrifugation is to centrifuge 15min in 4000r/min.
Beneficial effects of the present invention are：
（1）The present invention first uses BBD- response surface optimization methods founding mathematical models, obtain object function after, using genetic algorithm and
Direct search algorithm searches for the accurate optimal value of each factor, three kinds of methods is used in combination, to obtain optimum optimization as a result, overcoming
Limitation present in existing three kinds of methods.
（2）BBD- response surface optimizations method can intend complicated unknown function relationship in zonule with simple model
It closes, calculate simplicity, test number (TN) can be reduced, reduce cost etc., there is the advantage that can be continuously analyzed testing each level.It loses
Propagation algorithm has used for reference living nature natural selection and naturally hereditary mechanism, and initial data is made to generate heredity in an iterative process and become
It is different, and optimal experiment condition is looked for from global angle, it can effectively avoid subjectivity drawn game when optimal experiment condition is chosen
Portion's Optimal solution problem.Direct search algorithm can be accomplished that pattern search method as a supplement of other optimization methods,
Maximum, minimum positive group pattern, processing boundary constraint, linear equality, linear inequality are can search for out, and not needing function can be micro-
Or it is continuous.
（3）The extracting method optimized through the invention can be realized and be carried out to the greatest extent to Radix Glehniae saccharide compound
Extraction has important research significance.
Description of the drawings
Fig. 1 is single factor exploration result in embodiment 1.
Wherein, A is the recovery rate result of different grain size Radix Glehniae sample；B is carrying for different extraction time Radix Glehniae samples
Take rate result；C is the recovery rate result of different ratio of water to material Radix Glehniae samples；D is the extraction of different extraction time Radix Glehniae samples
Rate result.
Fig. 2 be in embodiment 1 extraction time and medicinal material granularity to the influence diagram of Radix Glehniae saccharide compound recovery rate.
Fig. 3 be in embodiment 1 ratio of water to material and medicinal material granularity to the influence diagram of Radix Glehniae saccharide compound recovery rate.
Fig. 4 be in embodiment 1 ratio of water to material and extraction time to the influence diagram of Radix Glehniae saccharide compound recovery rate.
Fig. 5 is that the optimal adaptation as obtained by genetic algorithm is write music line in embodiment 1.
Fig. 6 is pattern search result figure obtained by Direct search algorithm in embodiment 1.
Specific implementation mode
The testing program of the present invention is made further explanation below by specific embodiment.
Instrument and material used in the present invention
1, instrument
BT125D electronic balances（Sai Duolisi scientific instrument Co., Ltd）, spectrophotometry instrument UV-250（Shimadzu is public
Department）, DK-S24 type thermostat water baths（Shanghai is gloomy reliable to test Co., Ltd）, KDM-A control-temperature electric heating sets（Jintan City's instrument medical treatment
Factory）, DGG-9123A type drying boxes（Shanghai is gloomy reliable to test Instrument Ltd.）, FW-80 high speed Universal pulverizers（Beijing is forever
Bright Medical Instruments Co., Ltd）, TDL40B low speed centrifuges（Anting Scientific Instrument Factory, Shanghai）, pure water ultra-pure water all-in-one machine
（Mole company）.
2, material
Radix Glehniae medicinal material is purchased from the shops of Chinese medicines Jian Lian of Jinan City, and through Medicine Institute, Shandong Province, Lin Huibin researcher is accredited as umbrella shape
Section's plant glehnia littoralis（GLehnia Littoralis Fr.Schmidtex Mip.）Dry root；Glucose control product
（10010518）, anthrone and the concentrated sulfuric acid be purchased from Sinopharm Chemical Reagent Co., Ltd..
Embodiment 1
（1）The measurement of Radix Glehniae saccharide compound recovery rate
This experiment measures the recovery rate of Radix Glehniae saccharide compound using Anthrone-sulfuricacid method.
（2）The preparation of reagent
The preparation of anthrone reagent：It weighs 0.16g anthrones to be dissolved in the 80mL concentrated sulfuric acids, is slowly stirred and made it dissolve with glass bar,
It needed the same day to prepare when use to use.
（3）The selection of Detection wavelength and the foundation of glucose solution standard curve
105 DEG C of dryings are weighed to the DEXTROSE ANHYDROUS 0.5g of constant weight, is diluted with pure water and is settled to 100mL volumetric flasks, therefrom
It draws 1mL and is settled to 50mL volumetric flasks with pure water again, obtain the glucose standards solution of a concentration of 0.1mg/mL.It draws respectively
0.2,0.4,0.6,0.8,1.0,1.2mL glucose standards solutions enter the tool plug test tube of clean dried, every group of parallel 3 test tube,
It is 1.2mL that every test tube adds pure water to total liquid volume respectively, while 1.2mL pure water being taken to make blank control.
Each test tube is numbered in order, 4mL anthrone reagents are separately added into, it is rapid to immerse cooling in ice-water bath, shake well
Afterwards, each test tube immerses in boiling water bath simultaneously, and nozzle capping starts to react.Accurate response 10 minutes.Shake uniform, cooling, room temperature
It places after ten minutes, obtains standard sample.The length scanning of 400 ~ 900nm ranges is carried out using spectrophotometry instrument,
Obtain best detection wavelength.And under the Detection wavelength, the absorbance of each standard sample is measured.Each sample measure twice, with
The average value of absorbanceFor ordinate, with the concentration C of glucose（mg/mL）For abscissa, calculate glucose standard it is bent
Line equation, linear relationship and range of linearity result.
Length scanning result is it is found that best detection wavelength is 590nm.It calculates it is found that standard curve is Y=8.9463X+
0.0341,R 2 =0.9945,R=0.9972 > 0.995, illustrates that linear relationship is good, and the range of linearity be 16.82~
100.92ug/ml。
（4）The preparation of test solution
Precision weighs coastal glehnia root 5g, hot water return extraction, and extracting solution filtration after merging, centrifuges（4000r/min,
15min）, Aspirate supernatant is settled to 500mL volumetric flasks, and therefrom the accurate 4mL that measures is settled to 250mL volumetric flasks.It mixes well
Therefrom accurate afterwards to draw in the tool plug test tube that 1mL enters dried and clean, parallel 3 test tubes continue Anthrone Sulphuric acid reaction.Instead
Answer process same（3）In begin from " each test tube numbering in order, be separately added into 4mL anthrone reagents ", until " be measured sample "
Only.Then the absorbance of test solution is measured at 590 nm, and each sample measures twice, with the average value of absorbanceMeter
Calculate recovery rate result.
（5）The calculating of Radix Glehniae saccharide compound recovery rate
Radix Glehniae saccharide compound recovery rate（%）Calculation formula is：Recovery rate（%）The mass concentration * solution of=saccharide compound
The quality * 100 of volume * extension rates/Radix Glehniae medicinal material
（6）Single factor exploration
1. influence of the medicinal material granularity to Radix Glehniae saccharide compound recovery rate
Radix Glehniae medicine materical crude slice smashes it through different meshes（50,65,80,100,120,150,200 mesh）Sieve, obtain different grain size
Radix Glehniae sample.Under conditions of holding extraction time is 3h, extraction time is 1 time, ratio of water to material is 50mL/g constant, investigate
The influence of Radix Glehniae medicine materical crude slice and variant granularity Radix Glehniae sample to Radix Glehniae saccharide compound recovery rate.
Radix Glehniae medicine materical crude slice is directly extracted, gained recovery rate result is 21.7021%, hence it is evident that is less than variant granularity Radix Glehniae
The saccharide compound recovery rate of powder.The recovery rate result of variant granularity Radix Glehniae sample is by Fig. 1（A）It is shown.It is found that extraction
Rate reduces afterwards as the increase of medicinal material granularity first increases.When medicinal material granularity is 150 mesh, the recovery rate of Radix Glehniae saccharide compound
The reason of highest continues growing the granularity of medicinal material, and recovery rate reduces instead may be since coastal glehnia root granularity is too low so that
Extraction system viscosity becomes larger, and the recovery rate of saccharide compound reduces.Therefore for efficiently using Radix Glehniae, the granularity for choosing Radix Glehniae is
150 mesh are most suitable.
2. influence of the extraction time to Radix Glehniae saccharide compound recovery rate
Under conditions of holding ratio of water to material is 50mL/g, extraction time is 1 time, medicinal material granularity is that 150 purposes are constant, it is husky to measure north
Ginseng carries out single factor experiment in 1h, 2h, 3h, 4h, 5h, 6h recovery rate, with extraction time to recovery rate.
By Fig. 1（B）It is found that recovery rate then tends to be steady as the increase of extraction time first increases, it is in extraction time
Recovery rate starts to tend to be steady when 3h.The purpose for saving the energy is taken into account while ensuring that Radix Glehniae is efficiently utilized, so
Selection and withdrawal time 3h optimums.
3. influence of the ratio of water to material to Radix Glehniae saccharide compound recovery rate
Under conditions of holding medicinal material granularity is 150 mesh, extraction time is 1 time, extraction time is 3h constant, ratio of water to material is investigated
10,20,30,40,50, influences of the 60mL/g to Radix Glehniae saccharide compound recovery rate.
By Fig. 1（C）It is found that recovery rate then tends to be steady as the increase of ratio of water to material first increases, it is 50mL/ in ratio of water to material
Start to tend to be steady when g, it is that 50mL/g is most appropriate to choose ratio of water to material.
4. influence of the extraction time to Radix Glehniae saccharide compound recovery rate
Under conditions of to keep medicinal material granularity be 150 mesh, extraction time 3h, ratio of water to material are 50mL/g constant, north is investigated respectively
The root of straight ladybell hot water return extracts 1,2,3 recovery rate.（The recovery rate result of 2 gained of extraction time is accumulative to be extracted twice
The recovery rate result of the sum of recovery rate, 3 gained of extraction time is accumulative the sum of the recovery rate extracted three times）.
By Fig. 1（D）It is found that although recovery rate increases with the increase of extraction time, cumulative yield that extraction is 3 times with
2 cumulative yields only poor 0.8000773% of extraction, and there was no significant difference, it is contemplated that economic cost selective extraction 2 times is most suitable.
（6）The optimization analysis of BBD- response phase methods
1. factor and horizontal determination
On the basis of single factor experiment, using software Design-Expert 8.0.5b, with carrying for Radix Glehniae saccharide compound
It is response to take rate（Y）, medicinal material granularity is carried out using BBD- response phase methods（A）, extraction time（B）And ratio of water to material（C）Three factors
Three hydraulic tests.Factor is shown in Table 1 with level design.BBD test arrangements and it the results are shown in Table 2.
1 factor of table and water-glass
2 BBD- response phase methods design scheme of table and result
2. models fitting
Models fitting is carried out to each factor using software Design-Expert 8.0.5b.Software is recommended（Suggested）Mould
Type type is fitted for binomial equation（Quadratic）, fit equation is Y=- 69.04440+0.88849A+14.07365B+
0.49493C-0.040163AB-0.0000804375AC+0.055938BC-0.00232195A2-1.47066B2-
0.00669713C2（R 2=0.8915,P=0.0115）.It loses and intends errorP=0.6522, it is known that, the correlation of binomial fitting equation
Coefficient is higher, binomial modelP<0.05, it loses and intends errorP>0.05, illustrate that models fitting effect is preferable, therefore is carried out with the model
Analysis is reasonable.
3. variance analysis
The results of analysis of variance is shown in Table 3.
3 the results of analysis of variance of table
As shown in Table 3, B, A2It influences significantly, i.e., in three factors, extraction time B is relative to C pairs of ratio of water to material A and medicinal material granularity
The recovery rate of Radix Glehniae saccharide compound influences more notable.
Software is analyzed it is found that the optimum extraction process of Radix Glehniae saccharide compound is：Medicinal material granularity is 187.02 mesh, extraction
Time is 3.17h, solid-liquid ratio 49.04mL/g, can get the highest prediction extraction of Radix Glehniae saccharide compound with this condition
Rate result 38.402%.
4. response surface design figure and contour map
Response surface design figure and contour map have been obtained after software processing, Radix Glehniae saccharide compound has been extracted for analyzing each factor
The influence of rate, as a result as shown in Figure 2,3, 4：Response surface design figure and contour map can reflect the influence feelings of each factors on extraction rate
Condition.As shown in Figure 2, the shape of extraction time and medicinal material granularity contour is ellipse, illustrates extraction time and medicinal material granularity two
Because being known as certain reciprocation, extraction time and the curved surface gradient of medicinal material granularity are not much different, and the density difference of contour is not yet
Greatly, illustrate extraction time compared with two factor of medicinal material granularity, the influence to Radix Glehniae saccharide compound recovery rate is similar；By scheming
3 it is found that the shape of ratio of water to material and medicinal material granularity contour be ellipse, illustrate ratio of water to material and medicinal material granularity two because being known as certain friendship
The curved surface of interaction, ratio of water to material is more slow than the curved surface of medicinal material granularity, and the contour line density of medicinal material granularity is more than ratio of water to material, illustrates medicinal material
Influence bigger of the fineness ratio ratio of water to material to Radix Glehniae saccharide compound recovery rate；As shown in Figure 4, ratio of water to material and extraction time are contour
The shape of line is ellipse, illustrates ratio of water to material and extraction time two because being known as certain reciprocation, and the curved surface of ratio of water to material ratio carries
Take the curved surface of time slow, the density of extraction time contour is significantly greater than ratio of water to material, illustrates extraction time than medicinal material granularity to north
The extraction of the root of straight ladybell saccharide compound influences bigger.
（7）Genetic algorithm searching optimum extraction condition
Using BBD binomial fittings equation as object function, using the tool boxes GADS of software Matlab 2014b, calculated using heredity
Method searches for the optimum extraction condition of Radix Glehniae saccharide compound.Search process uses genetic algorithm tool GUI, parameter setting to be shown in Table
4.Random search 10 times, the results are shown in Table 5.Gained optimal adaptation line of writing music is shown in Fig. 5.
4 genetic algorithm tool GUI parameter settings of table
5 random search result of table
As shown in Figure 5, after about 50 generations of evolving, the optimal adaptation degree of Radix Glehniae saccharide compound recovery rate tends towards stability.And by
5 data statistic analysis of table is it is found that the range of variation of 10 random search results is smaller, to the approximation ratio of object function optimum value
Preferably.Therefore, this experiment selects the average level of 10 random searches, i.e. medicinal material granularity is 159.523377 mesh, extraction time
For 3.575010h, ratio of water to material is optimum extraction items of the 50.923100mL/g as genetic algorithm optimization Radix Glehniae saccharide compound
Part, it is contemplated that recovery rate result is 39.581479%.
（8）Direct search algorithm searches for optimum extraction condition
Using binomial fitting equation as object function, using the tool boxes GADS of software Matlab 2014b, using direct search
The optimum extraction condition of algorithm search Radix Glehniae saccharide compound.Search process use pattern research tool GUI, random search 10
Secondary, parameter setting is shown in Table 6.Gained pattern search result figure is shown in Fig. 6.
6 Direct search algorithm tool GUI parameter settings of table
It is found after Direct search algorithm random search 10 times, 10 times result is completely the same：It is 159.523438 mesh in medicinal material granularity,
Extraction time is 3.575012h, under conditions of ratio of water to material is 50.923096mL/g, it is contemplated that optimum extraction rate result is
39.581479%, and 10 search process stopped at for 74 generations.
（9）Result verification
According to actual conditions, the decimal digits of each extraction conditions is determined, it is known that the optimum results of BBD- response phase methods are：Medicinal material grain
Degree is 150 mesh, extraction time 3.2h, ratio of water to material 49mL/g.The best item optimized through genetic algorithm and Direct search algorithm
Part is not much different, and common optimum results are：Medicinal material granularity is 150 mesh, extraction time 3.6h, ratio of water to material 51mL/g.Point
Radix Glehniae saccharide compound, each condition parallel extraction 6 times are not extracted according to above two condition.Carry out statistics pairingtIt examines,
It is found that two groups of equal Normal Distributions of result, and two groups of results do not have significant difference（t=-1.021,P=0.354>0.05）,
I.e. use BBD- response phase methods, genetic algorithm and Direct search algorithm can preferably to the extraction of Radix Glehniae saccharide compound into
Row optimizing, and without significant difference.
Comparative example 1
If only with document《Response phase method optimizes the Study on extraction of carbohydrate in Radix Glehniae》The method of offer extracts item
Piece optimization, it is desirable that experimenter is accurately controlled each factor as possible, and the difficulty of technology controlling and process is made to increase.And it can not learn
The influence of the fluctuation pairing effect value of three factor of medicinal material granularity, extraction time and ratio of water to material.And obtained using Genetic algorithm searching
Optimal procedure parameters can fluctuate in a certain range, and have little influence on recovery rate, be more easily implemented in produce reality.This
Outside, Direct search algorithm can obtain the best of each technological parameter by setting boundary constraint, using pattern search method precise search
Value.Therefore, three kinds of methods are used in combination, and learn from other's strong points to offset one's weaknesses, and the optimum extraction process to obtain Radix Glehniae saccharide compound optimizes
As a result.
Claims (5)
1. a kind of extraction of the Radix Glehniae saccharide compound optimized using response phase method combination genetic algorithm and Direct search algorithm
Method, which is characterized in that include the following steps：
（1）The preparation of test solution：50-200 mesh coastal glehnia root 5g are weighed, are carried with ratio of water to material 10-60mL/g hot water returns
It takes 1-3 times, extraction time 1-6h, extracting solution filter after merging, centrifuges, Aspirate supernatant is settled to 500mL volumetric flasks, therefrom
Precision measures 4mL and is settled to 250mL volumetric flasks.It is therefrom accurate after mixing well to draw in the tool plug test tube that 1mL enters dried and clean,
Parallel 3 test tubes continue Anthrone Sulphuric acid reaction, then measure the absorbance of test solution, each sample at 590 nm
Product measure twice, with the average value of absorbanceCalculate recovery rate result；
（2）The making of standard curve：105 DEG C of dryings are weighed to the DEXTROSE ANHYDROUS 0.5g of constant weight, with pure water dilution and constant volume
To 100mL volumetric flasks, therefrom draws 1mL and be settled to 50mL volumetric flasks with pure water again, obtain the grape of a concentration of 0.1mg/mL
Standard for Sugars solution；Absorption 0.2,0.4,0.6,0.8,1.0,1.2mL glucose standards solutions respectively, every group of parallel 3 test tube, often
It is 1.2mL that branch test tube adds pure water to total liquid volume respectively, while 1.2mL pure water being taken to make blank control, is separately added into
4mL anthrone reagents, cooling in immersion ice-water bath rapidly, after shake well, each test tube immerses in boiling water bath simultaneously, and nozzle capping is opened
Beginning reaction, reacts 10 minutes, and shaking is uniform, cooling, is placed at room temperature for after ten minutes, obtains standard sample, and detect wave in 590nm
Under length, the absorbance of each standard sample is measured;Each sample measures twice, with the average value of absorbanceFor ordinate, with Portugal
The concentration C of grape sugar（mg/mL）For abscissa, the calibration curve equation of glucose is obtained；
（3）Experimental design and statistical analysis：1. single factor experiment：It is with medicinal material granularity, extraction time, ratio of water to material and extraction time
Parameter carries out single factor experiment, measures the saccharide compound content in Radix glehniae extract using Anthrone-sulfuricacid method, it is husky to calculate north
Join saccharide compound recovery rate（%）, calculation formula is：Recovery rate（%）The volume * of the mass concentration * solution of=saccharide compound is dilute
Release the quality * 100 of multiple/Radix Glehniae medicinal material；2. the optimization analysis of BBD- response phase methods：On the basis of single factor experiment, use
Software Design-Expert 8.0.5b, using the recovery rate of Radix Glehniae saccharide compound as response（Y）, during selection and withdrawal
The significant medicinal material granularity of influential effect（A）, extraction time（B）And ratio of water to material（C）Three factors are independent variable, establish polynary secondary returning
Return equation：Y=-69.04440+0.88849A+14.07365B+0.49493C-0.040163AB-0.0000804375AC+
0.055938BC-0.00232195A2-1.47066B2-0.00669713C2（R 2 =0.8915,P=0.0115）；Carry out software point
Analysis, obtains response surface design figure and contour map；
（4）Genetic algorithm searching is analyzed：Using BBD binomial fittings equation as object function, using software Matlab 2014b, adopt
With the optimum extraction condition of Genetic algorithm searching Radix Glehniae saccharide compound, search process uses genetic algorithm tool GUI, setting
Parameter, random search 10 times, verification analysis；
（5）Direct search method searching analysis：Using binomial fitting equation as object function, using software Matlab 2014b, adopt
With Direct search algorithm search for Radix Glehniae saccharide compound optimum extraction condition, search process use pattern research tool GUI,
Arrange parameter, random search 10 times, verification analysis.
2. extracting method according to claim 1, which is characterized in that step（4）In, the search process of the genetic algorithm
Using genetic algorithm tool GUI, arrange parameter is as follows：
。
3. extracting method according to claim 1, which is characterized in that step（5）In, the search of the direct search method
Journey use pattern research tool GUI, arrange parameter are as follows：
。
4. according to claim 1-3 any one of them extracting methods, which is characterized in that the method, which optimizes, is：Take north
The root of straight ladybell pharmaceutical decocting piece crushes as 150 mesh powder of granularity, and precision weighs Radix Glehniae medicinal powder, hot water return extraction, ratio of water to material
It for 49: 1 mL/g, is centrifuged after extracting solution filtration, retains supernatant, sediment fraction adds water to repeat last fetched process, adds water
Measure identical, each extraction time is 3.2h, is extracted 2 times altogether.
5. extracting method according to claim 4, which is characterized in that the condition of the centrifugation be in 4000r/min,
Centrifuge 15min.
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