CN108976309A - Polysaccharide extracting process and optimization of orthogonal test in passionflower leaf - Google Patents
Polysaccharide extracting process and optimization of orthogonal test in passionflower leaf Download PDFInfo
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- CN108976309A CN108976309A CN201810701321.2A CN201810701321A CN108976309A CN 108976309 A CN108976309 A CN 108976309A CN 201810701321 A CN201810701321 A CN 201810701321A CN 108976309 A CN108976309 A CN 108976309A
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- passionflower
- polysaccharide
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- passionflower leaf
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Abstract
The present invention is a kind of extracting method of polysaccharide in passionflower leaf with antioxidant activity.By the extraction process of Optimized by Orthogonal Test passionflower leaf polyose, the optimum extraction condition of passionflower leaf polyose is obtained.It the invention further relates to being applied in pharmacy, is especially applied in preparing anti-oxidation medicine, is included in and removes DPPH free radical drug, hydroxy radical drug and Fe3+The medicinal application of reducing power.Passionflower leaf polyose is used for antioxidation activity in vitro test by the present invention, and the application for passionflower leaf polyose in field of medicaments provides reference frame.
Description
Technical field
The present invention relates to a kind of optimization method of Polysaccharide from Bee in passionflower leaf, more particularly to utilizing orthogonal test
The method of method optimization passionflower leaf polyose extraction conditions.
Background technique
Passionflower (Passiflora edulis Sims) is Passifloraceae (Passiflraceae) Passiflora
(Passiflora) herbaceous species plant also known as passion fruit.Dahlia fruit juice have guava, pineapple, banana, mango, apple,
The fragrance of the pleasant of the various fruits such as smoked plum often takes its fruit juice for the processing such as beverage, candy.It is reported that passionflower has
Promote the production of body fluid to quench thirst, eliminate fatigue, refresh oneself sober up, skin-protecting face nursing, weight-reducing defaecation, clearing away the lungheat and moisturizing, preventing phlegm from forming and stopping coughing, enriching yin and nourishing kidney, anti-cancer
A variety of medicines and health protection functions such as anti-ageing, tranquilizing the mind antianxiety, the antibacterial, prevention and cure of cardiovascular disease of antalgic and inflammation relieving, thus also it is known as " fruit
In king ".
The extracting method of plant polyose is generally boiling waterbath, establishes continuous variable surface model, passes through regression equation
Come a series of relationship being fitted between multiple response variables and variables, investigation does not stop experimental factor grade reciprocation to target response
The influence of value.This method energy economical and effective, the optimum combination condition for quickly determining more experimental factors.This study tour difference
Solid-liquid ratio, extraction time and Extracting temperature obtain west kind using Responds Surface Methodology to the affecting laws of polysaccharide in passionflower leaf
The optimization technique of Polyose extraction in lotus leaf.
Summary of the invention
The purpose of the present invention is on the basis of single factor experiment, using orthogonal experiment to Polyose extraction in passionflower leaf
Technique extraction conditions optimize, and provide reference frame for the comprehensive development and utilization of passionflower.
To achieve the above object, the technical solution used in the present invention are as follows:
Step 1: fresh passionflower blade dries at 50 DEG C, crushed 20 meshes.According to certain solid-liquid ratio,
80% 85 DEG C of condensing refluxes of ethyl alcohol extract 3 times, each 2h, separate residue and dry to get removing lipid, pigment and small molecule
The powder of the passionflower leaf of impurity, it is spare.
Step 2: hot water extracts polysaccharide in passionflower leaf, and the change of Polyose extraction in passionflower leaf is determined using response phase method
Parameter is measured, the horizontal test method of three factor of setting five is carried out repeating to test obtaining experimental result, be calculated more in passionflower leaf
The recovery rate of sugar, then orthogonal test is used, it takes Three factors-levels to be analyzed, obtains optimum extraction condition: solid-liquid ratio 1:
35g/ml, extraction time 100min, 100 DEG C of temperature to be optimal, recovery rate is up to 3.006%.
Step 3: its absorbance is measured using phend-sulphuric acid, and calculates the recovery rate of polysaccharide in passionflower leaf.
The invention has the benefit that
The present invention determines the variable parameter of Polyose extraction in passionflower leaf using response phase method, chooses to more in passionflower leaf
Three factors that sugared recovery rate more significantly affects, i.e. solid-liquid ratio, extraction time and temperature, on the basis of single factor experiment, if
Count the orthogonal test of Three factors-levels, and then polysaccharide optimal extract process in optimization passionflower leaf.
Detailed description of the invention
Fig. 1 is influence of the solid-liquid ratio to polysaccharide extract rate in passionflower leaf
Fig. 2 is influence of the extraction time to polysaccharide extract rate in passionflower leaf
Fig. 3 is influence of the Extracting temperature to polysaccharide extract rate in passionflower leaf
Specific embodiment
By embodiment, technical scheme is described further with reference to the accompanying drawing.
1, sample preparation
It by the fresh blade drying of passionflower, crushes, is extracted 3 times, each 2h with 80% 85 DEG C of condensing refluxes of ethyl alcohol, separation
Residue and drying, it is spare.It weighs residue 5.0g in the balance, distilled water is added according to certain solid-liquid ratio, hot water extracts, and filters, filter
Liquid is centrifuged to obtain supernatant concentration, 4 times of volume dehydrated alcohol precipitatings is added, 4 DEG C of standings 48h, 4000r/min are centrifuged 15min, take
Drying is precipitated, Thick many candies in passionflower leaf are obtained.
2, experiment of single factor
Change solid-liquid ratio, extraction time, Extracting temperature progress experiment of single factor, is mentioned using phend-sulphuric acid quantitative determination
Polyoses content in liquid is taken, the recovery rate of polysaccharide in passionflower leaf is calculated, experiment is parallel three times every time.
3, optimization of orthogonal test designs
According to experiment of single factor as a result, choosing the significant solid-liquid ratio of Polyose extraction influential effect, extraction time, Extracting temperature
These three factors, are tested using Orthogonal Experiment and Design, are chosen solid-liquid ratio, extraction time and Extracting temperature and are carried out experimental factor and water
Flat setting, setting the results are shown in Table 1.
1 experimental factor of table and level set
With solid-liquid ratio (A), extraction time (B) and Extracting temperature (C), using the recovery rate of polysaccharide as response (Y), test side
Case and it the results are shown in Table 2.
4, test result analysis and optimization
According to orthogonal experiments it is found that passionflower leaf polyose extracts Optimal Parameters are as follows: solid-liquid ratio 1:35g/mL, when extraction
Between 100min, 100 DEG C of Extracting temperature.With this condition, polysaccharide extract rate is 3.006% in passionflower leaf and recovery rate is relatively inclined
Poor smaller, stable process conditions.
5, influence of the different passionflower leaf polyose concentration to DPPH free radical scavenging ability
The analyte sample fluid 2.0mL prepared is measured, 2.0mL 0.04mg/mL DPPH solution is added, and (dehydrated alcohol is as molten
Agent), 30min is protected from light after mixing well at room temperature, its absorbance A i is measured at wavelength 517nm, while measuring anhydrous second
The absorbance A C of alcohol (2.0ml) and DPPH (2.0mL) mixed liquor, dehydrated alcohol (2.0mL) and sample liquid (2.0mL) mixed liquor
Absorbance A j.
Doses effect is presented to the scavenging effect of DPPH free radical in passionflower leaf polyose in concentration range, and with
The increase of concentration and improve, when concentration reaches 1.0mg/mL, passion flower-fruit peel polysaccharide is to the clearance rate of DPPH
23.09%.
6, influence of the different passionflower leaf polyose concentration to hydroxyl radical free radical Scavenging activity
Take the FeSO of 0.2mL4α-deoxyribose of 0.2mL is added in test tube with ground stopper in-EDTA mixed liquor (10mmol/L)
Solution (20mmol/L) then adds 0.2mL sample, and is settled to phosphate buffer (0.2mol/L, pH=7.4)
Then the H of 0.2mL is added in 1.8mL2O2(10mmol/L), 40 DEG C of water bath with thermostatic control 1h, the trichloroacetic acid that 1mL 2.8% is added terminate
Reaction, adds 1% thiobarbituricacidα- of 1mL, heats 10min after mixing in boiling water bath, surveys at 532nm after cooling
Absorbance value AS.
Passionflower leaf polyose improves the Scavenging activity of OH with the increase of concentration, when concentration reaches 1.0mg/mL
When, passion flower-fruit peel polysaccharide clearance rate is 31.23%.
7, different passionflower leaf polyose concentration are to Fe3+Reducing power effect
0.5ml sample liquid is measured in test tube, sequentially adds the 1%K of 0.5ml3Fe(CN)6Solution and 0.5ml PBS solution
(0.2mo1/L, pH=6.7) is cooled down immediately after 50 DEG C of water-bath 20min, sequentially adds 0.5ml 10%TCA solution,
0.5ml 0.1%FeCl3Solution and 2.0ml deionized water, sufficiently shake up, and stand 10min, measurement is shaken up at wavelength 700nm
Its absorbance value A, using ascorbic acid as positive control.
Doses effect is presented to reproducibility ability scavenging effect in passionflower leaf polyose in concentration range, and with dense
The increase of degree and improve, when concentration reaches 1.0mg/mL, passion fruit stem polysaccharide is to the clearance rate of reproducibility ability
22.70%.
Claims (6)
1. a kind of preparation method with polysaccharide in antioxidant activity passionflower leaf, it is characterised in that: the following steps are included:
(1) selection of raw material: choosing fresh passionflower blade, removes corruption, rotten part, and wash clean is spare；
(2) dry: the passionflower blade that step (1) is selected being put under 70-80 DEG C of oven drying 12-16h or sunlight and is dried；
(3) alcohol extracting: passionflower blade that step (2) are dried is 1:20g/mL according to solid-liquid ratio, and 85 DEG C of ethyl alcohol 80% are cold
Solidifying refluxing extraction 3 times, each 2h separates residue and drying to get the passionflower leaf for removing lipid, pigment and small molecular weight impurity
Powder, it is spare；
(4) water mentions: the passionflower leaf that step (3) alcohol extracting obtains is determined polysaccharide optimum extraction in passionflower leaf with orthogonal experiment
Condition, solid-liquid ratio 1:35g/mL, extraction time 100min, 100 DEG C of temperature；
(5) alcohol precipitation: will be put into refrigerator after the Aqueous extracts heating concentration of step (4), the dehydrated alcohol of 4 times of volumes be added after cooling,
After being sufficiently stirred, 4 DEG C of preservations are stood overnight, and precipitation solution is centrifuged 15min at 4000r/min, and taking precipitate is freeze-dried,
Obtain polysaccharide in passionflower leaf.
2. polysaccharide in passionflower leaf according to claim 1, which is characterized in that polysaccharide answering in pharmaceutical purpose in passionflower leaf
With.
3. polysaccharide in passionflower leaf according to claim 1, which is characterized in that in passionflower leaf polysaccharide prepare it is anti-oxidant
It is applied in drug.
4. the application of polysaccharide in passionflower leaf according to claim 2, which is characterized in that in passionflower leaf prepared by polysaccharide
The application in DPPH free radical drug is removed, polysaccharide effective concentration is 1mg/mL in passionflower leaf.
5. the application of polysaccharide in passionflower leaf according to claim 2, which is characterized in that in passionflower leaf prepared by polysaccharide
The application in hydroxyl radical free radical drug is removed, polysaccharide effective concentration is 1mg/mL in passionflower leaf.
6. the application of polysaccharide in passionflower leaf according to claim 3, which is characterized in that in passionflower leaf prepared by polysaccharide
The application in reducing power drug is being removed clearly, polysaccharide effective concentration is 1mg/mL in passionflower leaf.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111825780A (en) * | 2020-08-27 | 2020-10-27 | 桂林理工大学 | Preparation method and degradation process of passion flower pericarp enzyme-assisted alkali extraction polysaccharide |
| CN111961698A (en) * | 2020-08-27 | 2020-11-20 | 桂林理工大学 | Preparation method and application of passion fruit peel polysaccharide degraded by lactobacillus brevis enzyme production |
| CN112979839A (en) * | 2021-04-15 | 2021-06-18 | 桂林理工大学 | Preparation method of passion flower leaf acidic polysaccharide |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111825780A (en) * | 2020-08-27 | 2020-10-27 | 桂林理工大学 | Preparation method and degradation process of passion flower pericarp enzyme-assisted alkali extraction polysaccharide |
| CN111961698A (en) * | 2020-08-27 | 2020-11-20 | 桂林理工大学 | Preparation method and application of passion fruit peel polysaccharide degraded by lactobacillus brevis enzyme production |
| CN112979839A (en) * | 2021-04-15 | 2021-06-18 | 桂林理工大学 | Preparation method of passion flower leaf acidic polysaccharide |
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Application publication date: 20181211 |