CN109010409A - The extraction and purification process and detection method of content of flavonoid compound in a kind of people face fruit leaf - Google Patents
The extraction and purification process and detection method of content of flavonoid compound in a kind of people face fruit leaf Download PDFInfo
- Publication number
- CN109010409A CN109010409A CN201810820608.7A CN201810820608A CN109010409A CN 109010409 A CN109010409 A CN 109010409A CN 201810820608 A CN201810820608 A CN 201810820608A CN 109010409 A CN109010409 A CN 109010409A
- Authority
- CN
- China
- Prior art keywords
- solution
- general flavone
- people face
- reference substance
- content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/38—Clusiaceae, Hypericaceae or Guttiferae (Hypericum or Mangosteen family), e.g. common St. Johnswort
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
Landscapes
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides general flavone enrichment and purification methods in a kind of people face fruit leaf.By 60%~90% ethyl alcohol heating and refluxing extraction of people face fruit leaf, after being concentrated under reduced pressure to give crude extract, upper polyamide column purifying is to get sample after purification.The present invention measures people face fruit leaf flavonoids content using ultraviolet-visible spectrophotometry, using the morelloflavone (fukugetin) and fukugiside (fukugiside) content in high effective liquid chromatography for measuring general flavone.The raw material that the present invention uses is people face fruit leaf, low in cost, general flavone content after purification is stablized, and utilizes ultraviolet spectrophotometry detection and high performance liquid chromatography detection, its content can reach 50% or more, can satisfy requirement of five kind new medicine of Chinese medicine to active component content.
Description
Technical field
The present invention relates to general flavone enriching and purifying technique in plant pharmaceutical technology field more particularly to a kind of people face fruit leaf with
And in corresponding extract Flavonoid substances detection method of content.
Background technique
People face fruit (Garcinia xanthochymus Hook.f.) is Guttiferae gamboge platymiscium.In Thailand and India
In traditional medicine, the fruit of people face fruit can treat disease in the liver and gallbladder, diarrhea and dysentery.As traditional dai medicine, people face fruits and
Leaf has expelling parasite, clearing heat and detoxicating, solution food poisoning and other effects.Early period is phytochemical to its leaf studies have shown that mainization
It studies point based on flavone compound, morelloflavone (fukugetin) and fukugiside (fukugiside) are that it is representative
Compound, and show good alpha amylase inhibitory activity and GSK-3 β inhibitory activity, and there is anti-diabetic, anti-oxidant
With anti-inflammatory a variety of pharmacological activity, general flavone in the fruit leaf of people face is subjected to enriching and purifying, it is potential to develop a kind of novel health sugar
Urinate medicine.People face fruit leaf be integration of drinking and medicinal herbs medicinal material, leaf extract can reduce existing marketed drugs alimentary canal stimulation and
The common side reaction such as dizzy palpitaition has the general flavone of phenolic hydroxyl group abundant in the fruit leaf extract of people face, can multi-path adjust
Blood glucose, while there is antioxidant activity, diabetic polyneuropathy can be significantly improved, alleviates diabetes to a certain extent
Pathology occurrence and development process.Morelloflavone (fukugetin) in the fruit leaf of people face has preferable anti-diabetic activity, anti-oxidant
Activity, anti-tumor activity, anti-diabetic activity.
To the technical study that general flavone in the fruit leaf of people face isolates and purifies, it is particularly important in new drug development.It is Chinese special
A kind of system of the extract of people face fruit leaf is disclosed in the application of sharp CN106074636A people face fruit leaf extract and preparation method
Preparation Method, after including the following steps: (1) dry people face fruit leaf pulverizing medicinal materials, the second for being 40%-80% with concentration of volume percent
Alcohol solution extracts, and after extracting solution filters and recycles ethyl alcohol, obtains the alcohol extract of people face fruit leaf；(2) alcohol extract of people face fruit leaf adds
Water adjusts concentration, pH to 7~8 is adjusted, after upper HPD100 large pore resin absorption column absorption, with water elution, then with volume basis
The ethanol water that specific concentration is 20~30% elutes, then the ethanol aqueous wash for being 50~70% with concentration of volume percent
It is de-, the ethanol water elution liquid of 50-70% is collected, is recovered under reduced pressure to no alcohol taste, arrives HPD100 resin eluent, it is dry, i.e.,
Obtain the extractive of general flavone of people face fruit leaf.Wherein contain 3 kinds of flavone compounds, is respectively as follows: good fortune wood glucoside, Fukugetin, good fortune tree
Flavine Fukugiside and bilobetin.And the quality sum of 3 kinds of compounds should be not less than total flavone part quality
50%.For this 3 kinds of compounds through high effective liquid chromatography for measuring, the sum of gross mass accounts for the 55% of people face fruit leaf flavonoids position.
(3) extractive of general flavone of people face fruit leaf is taken to dissolve with 50~60 DEG C of water, after adjusting concentration, upper 30-60 mesh polyamide column, first
Be eluted with water, then successively use concentration of volume percent 20%, 35%, 60% and 75% ethanol elution, respectively collect 35%,
60% and 75% ethanol eluate, is recovered under reduced pressure to a small amount of volume, then be freeze-dried, and respectively obtains compound good fortune wood glucoside, good fortune
Set flavine and bilobetin.
However, having the disadvantage in that the technique is always yellow using HPD100 macroporous adsorbing resin for purification in above-mentioned preparation method
Ketone, in loading with 10% NaHCO3Adjusting pH is 8, and extracting solution is in alkaline medium, due to the biflavone in the fruit of people face
Class compound, there is a flavanone structural unit, and flavanone open loop easily in lye is transformed into corresponding chalcone knot
Structure unit, may cause and be enriched to general flavone is not unborn flavone compound in plant, but chemical derivatization goes out
The compound come.
Summary of the invention
The present invention solves above-mentioned deficiency in the prior art, provides a kind of people face fruit leaf flavonoids enriching and purifying work
Skill and general flavone detection method and morelloflavone and fukugiside detection method of content.This method simple process, significant effect,
It is suitable for technology production requirement, high financial profit.
Realize technical solution used by above-mentioned purpose of the present invention are as follows:
The extraction and purification process of flavonoid compound in a kind of people face fruit leaf, comprising the following steps:
(1) 6~8 times of volumes of people face fruit leaf raw material are taken, the ethanol solution that volumetric concentration is 60%~90% heats back
Stream extracts, and obtains extracting solution；
(2) extracting solution is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting thing；
(3) 80~100 mesh polyamide columns in gained alcohol extracting thing, alcohol extracting thing: polyamide amount=1:2, diameter are walked on
High ratio=1:3, flow velocity=0.5BV/h successively elute 5~7 times of column volumes with 25~35% ethanol solution of volumetric concentration, use volume
45~55% ethanol solution of concentration elutes 9~11 times of column volumes, and 65~75% ethanol solution of volumetric concentration elutes 5~7 times of cylinders
Product, 65~75% ethanol solution of third column volume eluent and volumetric concentration of 45~55% ethanol solution of collected volume concentration
Second column volume eluent, merge two parts eluent, be recovered under reduced pressure ethyl alcohol and be freeze-dried after resulting dried powder
As extractive of general flavone detects general flavone content in extractive of general flavone, and content is more than 50%.
It is 80 DEG C that temperature is heated to reflux in step (1), and the time is 1 hour.
6 times of column volumes successively are eluted with 30% ethanol solution of volumetric concentration in step (3), it is molten with 50% ethyl alcohol of volumetric concentration
Liquid elutes 10 times of column volumes, and 70% ethanol solution of volumetric concentration elutes 6 times of column volumes.
General flavone content in extractive of general flavone can be examined using ultraviolet-visible spectrophotometry in step (3)
It surveys, comprising the following steps:
(1) preparation of reference substance solution: taking morelloflavone reference substance, accurately weighed, is dissolved with methanol, is configured to every
The reference substance solution of 1mL solution reference substance containing morelloflavone 0.1mg；
(2) preparation of test solution: being taken extractive of general flavone accurately weighed, dissolved with ethyl alcohol, and it is molten to be configured to every 1mL
The test solution of liquid 0.25mg containing sample；
(3) preparation of standard curve: precision measures reference substance solution 0.5mL, 1.0mL, 1.5mL, 2.0mL, 2.5mL points
It is not placed in 10mL volumetric flask, respectively plus methanol is to 3mL, adds the acetic acid of 0.1mol/L aluminium chloride ethanol solution 2mL plus pH 5.2
Sodium/acetate buffer solution 3mL, is finally settled to scale with ethyl alcohol, using corresponding reagent as blank, shakes up, and places at 35 DEG C
15min, then to measure absorbance at the wavelength of 415nm, using absorbance as ordinate, quality is abscissa, and it is bent to draw standard
Line；
(4) measuring method: precision measures test solution 1mL, is placed in 10mL measuring bottle, according to the measurement side in step (3)
Method measure absorbance, and then from standard curve calculate test solution in morelloflavone amount；
(5) measurement result: from regression equation calculation general flavone content.
Also general flavone content in extractive of general flavone is detected using high performance liquid chromatography in step (3), including
Following steps:
(1) preparation of reference substance solution: since functional component is mainly morelloflavone and good fortune wood in extractive of general flavone
Glycosides, therefore precision weighs morelloflavone reference substance 20mg, fukugiside reference substance 10mg respectively first, adds 95% chromatography methanol molten
Solution, is configured to the reference substance solution of every mL 0.4mg containing morelloflavone, the 0.2mg containing fukugiside；
(2) preparation of test solution: precision weighs extractive of general flavone 20mg, fixed after the dissolution of 95% chromatography methanol
Hold to 50mL, obtaining concentration is 0.4mg/mL test solution；
(3) sample measures: reference substance and test sample are measured with high performance liquid chromatograph, chromatographic condition are as follows: chromatographic column:
Agilent C18 chromatographic column (4.6mm ' 250mm, 5 μm)；Detection wavelength: 289nm；Flow velocity: 1.0mL/min；Column temperature: 30 DEG C；Into
Sample amount: 10 μ L；Mobile phase: acetonitrile (A), methanol (B), 0.5% phosphate aqueous solution (C), gradient elution, elution program are as follows:
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) | Mobile phase C (%) |
| 0 | 15 | 20 | 65 |
| 15 | 15 | 20 | 65 |
| 20 | 20 | 25 | 55 |
| 40 | 20 | 25 | 55 |
(4) result calculates: according to the chromatogram of sample and reference substance, recording peak area, is calculated in sample using external standard method
The content of morelloflavone and fukugiside.
Technical solution provided by the invention having the beneficial effect that compared with the prior art
(1) preparation method of the present invention, simple process, significant effect are suitable for technology production requirement, same to patent
CN106074636A is compared, and avoids having used alkaline medium, while the technological parameter applied sample amount of purifying has been determined, diameter height ratio and stream
Speed.
(2) preparation method of the present invention, solvent for use are ethanolic aqueous system, be normal drug production in commonly use it is molten
Agent, property safe to the human body is good, and is easily recycled, high financial profit.
(3) under the premise of guaranteeing general flavone content and yield, using 50% and 70% Fractional Collections, while making 50% second
Alcohol has impurity removal function concurrently, makes in 50% and 70% elution fraction that some general flavone content reaches 50% or more respectively.
(4) it compared with patent CN106074636A, is measured the present invention provides a kind of using ultraviolet-visible spectrophotometry
The method of people face fruit leaf extract general flavone content, method specificity is strong, accurate simple, with traditional with rutin or Quercetin
Method for the determination of total flavonoids of reference substance compares, and can effectively detect people face fruit leaf flavonoids content.Pass through height simultaneously
Effect liquid phase chromatogram develops a kind of new chromatographic condition, at the same measure anti-diabetic in general flavone active material morelloflavone and
The content of fukugiside.
(5) by studying people face fruit leaf total flavone extract, a kind of preparation of people face fruit leaf extract is established
And detection method, be conducive to the R and D of people face fruit leaf Related product.
Detailed description of the invention
Fig. 1 is extraction process flow chart provided by the present invention；
Fig. 2 is the high-efficient liquid phase chromatogram of extractive of general flavone obtained in the embodiment of the present invention.
Specific embodiment
Detailed specific description is done to the present invention combined with specific embodiments below.
Embodiment 1
The preparation process flow of general flavone sample as shown in Figure 1, it is specific the preparation method is as follows:
People face fruit leaf 2000g after taking drying and crushing, is added 70% ethyl alcohol of 12L, the refluxing extraction 1 under the conditions of 80 DEG C
Hour, it extracts once, is filtered under diminished pressure, extracting solution is then concentrated into no alcohol taste, upper 4.0kg polyamide chromatographic column (column diameter
20cm, pillar height 60cm, 1BV=10L), flow velocity=0.5BV/h is purified.Elution requirement are as follows: 30% 6 times of ethanol elution cylinder
Product；50% 10 times of ethanol elution column volume；70% ethyl alcohol elutes 6 times of column volumes respectively.Collect 50% and 70% ethanol eluate
Middle content the best part, is recovered under reduced pressure ethyl alcohol, and freeze-drying obtains dried powder, obtains 110.4g extractive of general flavone.
Embodiment 2
Determination of total flavonoids, it is double yellow with gamboge using visible spectrophotometry (2015 editions Chinese Pharmacopoeia general rules 0401)
Ketone (self-control, purity 98%) is reference substance, is reacted according to flavone compound with aluminium salt and generates yellow complex, measures general flavone
Content, specific experimental method is as follows:
The preparation of 1 reference substance solution
Take morelloflavone Fukugetin reference substance appropriate, it is accurately weighed, it is dissolved with methanol, is configured to every 1mL containing gamboge
The reference substance solution of biflavone Fukugetin reference substance 0.1mg to get.
The preparation of 2 test solutions
Take 1 drying sample powder of example appropriate, it is accurately weighed, it is dissolved with ethyl alcohol, is configured to every 1mL 0.25mg containing sample
Test solution to get.
3 Determination of Total Flavonoids
Precision measures reference substance solution 0.5mL, 1.0mL, 1.5mL, 2.0mL, 2.5mL and is respectively placed in 10mL volumetric flask,
Respectively plus methanol is to 3mL, adds the acetate/acetic buffer 3mL of 0.1mol/L aluminium chloride ethanol solution 2mL plus pH 5.2, finally
It is settled to scale with ethyl alcohol, using corresponding reagent as blank, is shaken up, places 15min at 35 DEG C.According to ultraviolet-visible spectrophotometry
(2015 editions general rules 0401 of Chinese Pharmacopoeia), to measure absorbance at the wavelength of 415nm.Using absorbance as ordinate, quality (mg)
For abscissa, standard curve is drawn.Precision measures test solution 1mL, is placed in 10mL measuring bottle, under sighting target directrix curve preparation
Method measure absorbance in accordance with the law from " adding methanol to 3mL ".It is double yellow from gamboge in test solution is calculated on standard curve
The amount of ketone Fukugetin, calculate to get.
As a result: this product is calculated by dry product, containing general flavone in terms of morelloflavone Fukugetin (C30H20O11), is
55.24%.
Embodiment 3
HPLC method surveys content, is morelloflavone (fukugetin) and its glycosides good fortune in view of people face berry extract effective component
The wooden glycosides (fukugiside), utilizes the two compounds contents in high effective liquid chromatography for measuring after purification sample, specific experiment
Method is as follows:
1 chromatographic condition
Chromatographic column: Agilent C18 chromatographic column (4.6mm × 250mm, 5 μm)；Detection wavelength: 289nm；Flow velocity:
1.0mL/min；Column temperature: 30 DEG C；Sample volume: 10 μ L；Mobile phase: acetonitrile (A), methanol (B), 0.5% phosphate aqueous solution (C), ladder
Degree elution, elution program see the table below 1, and sample chromatogram figure is shown in Fig. 2.
The preparation of 2 test samples
Sample drying powder about 20mg, accurately weighed in Example 1, after the dissolution of 95% chromatography methanol, is settled to
50mL, obtaining concentration is 0.4mg/ml test sample.
The preparation of 3 reference substances
Precision weighs morelloflavone reference substance 20mg, fukugiside reference substance 10mg respectively, and 95% chromatography methanol is added to dissolve,
It is configured to the reference substance solution of every mL 0.4mg containing morelloflavone, the 0.2mg containing fukugiside.
4 assays
Accurate reference substance mother liquor 1.0mL, 2.0mL, 3.0mL, 4.0mL, 5.0mL of drawing is placed in 10mL volumetric flask respectively,
With 95% methanol constant volume to scale.10 μ L are respectively drawn again, are measured by following chromatographic condition sample introductions.
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) | Mobile phase C (%) |
| 0 | 15 | 20 | 65 |
| 15 | 15 | 20 | 65 |
| 20 | 20 | 25 | 55 |
| 40 | 20 | 25 | 55 |
Table 1
Linear regression is carried out to concentration X (mg/mL) with peak area Y, obtains morelloflavone and fukugiside regression equation difference
For Y=460.22X-0.5378 (r=0.999) and Y=308.64X-0.148 (r=0.999).Test sample is pressed into similarity condition
Sample detection measures peak area substitution regression equation and finds out concentration, and then calculates respective content.HPLC chromatogram is as shown in Figure 2.
As a result: morelloflavone content is 33.32%, and fukugiside content is 19.17%, sum of the two 52.49%.
The foregoing is only a preferred embodiment of the present invention, single, scope of protection of the present invention is not limited thereto,
It should be understood by those skilled in the art that the above embodiments do not limit the invention in any form, it is all to use equivalent replacement or wait
The mode technical solution obtained for imitating transformation, falls within the scope of protection of the present invention.
Claims (5)
1. the extraction and purification process of flavonoid compound in a kind of people face fruit leaf, which comprises the following steps:
(1) 6~8 times of volumes of people face fruit leaf raw material are taken, the ethanol solution that volumetric concentration is 60%~90% is heated to reflux and mentions
It takes, obtains extracting solution；
(2) extracting solution is concentrated under reduced pressure into no alcohol taste, obtains alcohol extracting thing；
(3) 80~100 mesh polyamide columns in gained alcohol extracting thing, alcohol extracting thing: polyamide amount=1:2, the high ratio of diameter are walked on
=1:3, flow velocity=0.5BV/h successively elute 5~7 times of column volumes with 25~35% ethanol solution of volumetric concentration, use volumetric concentration
45~55% ethanol solutions elute 9~11 times of column volumes, and 65~75% ethanol solution of volumetric concentration elutes 5~7 times of column volumes, receive
Collect the of 65~75% ethanol solution of third column volume eluent and volumetric concentration of 45~55% ethanol solution of volumetric concentration
Two column volume eluents merge two parts eluent, and resulting dried powder is after ethyl alcohol is recovered under reduced pressure and is freeze-dried
Extractive of general flavone detects general flavone content in extractive of general flavone, and content is more than 50%.
2. the extraction and purification process of flavonoid compound in people face fruit leaf according to claim 1, it is characterised in that: step
(1) it is 80 DEG C that temperature is heated to reflux in, and the time is 1 hour.
3. the extraction and purification process of flavonoid compound in people face fruit leaf according to claim 1, it is characterised in that: step
(3) 6 times of column volumes successively are eluted with 30% ethanol solution of volumetric concentration in, elute 10 times of columns with 50% ethanol solution of volumetric concentration
Volume, 70% ethanol solution of volumetric concentration elute 6 times of column volumes.
4. the extraction and purification process of flavonoid compound in people face fruit leaf according to claim 1, it is characterised in that: step
(3) general flavone content in extractive of general flavone is detected using ultraviolet-visible spectrophotometry in, comprising the following steps:
(1) preparation of reference substance solution: taking morelloflavone reference substance, accurately weighed, is dissolved with methanol, it is molten to be configured to every 1mL
The reference substance solution of liquid reference substance containing morelloflavone 0.1mg；
(2) preparation of test solution: being taken extractive of general flavone accurately weighed, dissolved with ethyl alcohol, is configured to every 1mL solution containing sample
The test solution of product 0.25mg；
(3) preparation of standard curve: precision measures reference substance solution 0.5mL, 1.0mL, 1.5mL, 2.0mL, 2.5mL and is respectively placed in
In 10mL volumetric flask, respectively plus methanol is to 3mL, adds the acetate/acetic of 0.1mol/L aluminium chloride ethanol solution 2mL plus pH5.2 slow
Fliud flushing 3mL, is finally settled to scale with ethyl alcohol, using corresponding reagent as blank, shakes up, and 15min is placed at 35 DEG C, then with
Absorbance is measured at the wavelength of 415nm, using absorbance as ordinate, quality is abscissa, draws standard curve；
(4) measuring method: precision measures test solution 1mL, is placed in 10mL measuring bottle, surveys according to the measuring method in step (3)
Determine absorbance, and then from the amount for calculating morelloflavone in test solution on standard curve；
(5) measurement result: from regression equation calculation general flavone content.
5. the extraction and purification process of flavonoid compound in people face fruit leaf according to claim 1, it is characterised in that: step
(3) general flavone content in extractive of general flavone is detected using high performance liquid chromatography in, comprising the following steps:
(1) preparation of reference substance solution: since functional component is mainly morelloflavone and fukugiside in extractive of general flavone, because
Precision weighs morelloflavone reference substance 20mg, fukugiside reference substance 10mg respectively first for this, adds 95% chromatography methanol to dissolve, matches
The reference substance solution of every mL 0.4mg containing morelloflavone, the 0.2mg containing fukugiside is made；
(2) preparation of test solution: precision weighs extractive of general flavone 20mg, after the dissolution of 95% chromatography methanol, is settled to
50mL, obtaining concentration is 0.4mg/mL test solution；
(3) sample measures: measuring reference substance and test sample, chromatographic condition are as follows: chromatographic column: Agilent with high performance liquid chromatograph
C18 chromatographic column (4.6mm × 250mm, 5 μm)；Detection wavelength: 289nm；Flow velocity: 1.0mL/min；Column temperature: 30 DEG C；Sample volume: 10
μL；Mobile phase: acetonitrile (A), methanol (B), 0.5% phosphate aqueous solution (C), gradient elution, elution program are as follows:
(4) result calculates: according to the chromatogram of sample and reference substance, recording peak area, calculates gamboge in sample using external standard method
The content of biflavone and fukugiside.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810820608.7A CN109010409B (en) | 2018-07-24 | 2018-07-24 | Extraction and purification process and content detection method of total flavonoids in herminium leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810820608.7A CN109010409B (en) | 2018-07-24 | 2018-07-24 | Extraction and purification process and content detection method of total flavonoids in herminium leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109010409A true CN109010409A (en) | 2018-12-18 |
| CN109010409B CN109010409B (en) | 2021-05-14 |
Family
ID=64644805
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810820608.7A Active CN109010409B (en) | 2018-07-24 | 2018-07-24 | Extraction and purification process and content detection method of total flavonoids in herminium leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109010409B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113436A (en) * | 2021-12-02 | 2022-03-01 | 广州白云山光华制药股份有限公司 | Method for measuring total flavone content of scutellaria baicalensis based on fingerprint spectrum and application |
| CN115508480A (en) * | 2022-08-09 | 2022-12-23 | 北京康华远景科技股份有限公司 | Detection method of total flavonoids in dandelion plant and extract thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106074636A (en) * | 2016-07-19 | 2016-11-09 | 中南民族大学 | The application of people face fruit leaf extract and preparation method |
| CN107198705A (en) * | 2017-06-16 | 2017-09-26 | 江苏天晟药业股份有限公司 | A kind of people face berry extract and its preparation method and application |
-
2018
- 2018-07-24 CN CN201810820608.7A patent/CN109010409B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106074636A (en) * | 2016-07-19 | 2016-11-09 | 中南民族大学 | The application of people face fruit leaf extract and preparation method |
| CN107198705A (en) * | 2017-06-16 | 2017-09-26 | 江苏天晟药业股份有限公司 | A kind of people face berry extract and its preparation method and application |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113436A (en) * | 2021-12-02 | 2022-03-01 | 广州白云山光华制药股份有限公司 | Method for measuring total flavone content of scutellaria baicalensis based on fingerprint spectrum and application |
| CN115508480A (en) * | 2022-08-09 | 2022-12-23 | 北京康华远景科技股份有限公司 | Detection method of total flavonoids in dandelion plant and extract thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109010409B (en) | 2021-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101863871B (en) | Total glycosides of Rhodiola rosea, medical application and preparation method thereof | |
| CN102641326B (en) | Astragalus extract, as well as preparation and application methods thereof | |
| CN103070912B (en) | Sophora flavescens totall flavone extract product, preparation method and quality detection method | |
| CN101851261A (en) | Polygonum perfoliatum medicinal material, method for preparing reference substance of active constituents in preparation thereof as well as content determination method | |
| CN104130226B (en) | A kind of preparation method of high-content salvianolic acid B from salvia miltiorrhiza | |
| CN103969350B (en) | A kind of method for building up of antiphlogistic Chunwang capsule HPLC finger printing | |
| CN103340938A (en) | Preparation method for tartary buckwheat standard extract FT83 and high pure quercetin | |
| CN109010409A (en) | The extraction and purification process and detection method of content of flavonoid compound in a kind of people face fruit leaf | |
| CN108670973A (en) | A kind of Chloranthus glaber anti-influenza virus activity extract and preparation method thereof | |
| CN108113985B (en) | Application of eupatorium lindleyanum flavone part in preparation of anti-hepatitis B virus medicines | |
| CN103040945B (en) | Preparation and detection method of gangsong general flavones | |
| CN111595972A (en) | Preparation method and quality detection method of lithocarpus polystachyus rehd total flavone extract | |
| CN103940942B (en) | A kind of detection method of CHANGYANNING preparation | |
| CN102204999A (en) | Method for measuring content of luteolin in callicarpa nudiflora preparation | |
| Yu et al. | Simultaneous determination of six active compounds in Yixin Badiranjibuya granules, a traditional Chinese medicine, by RP-HPLC-UV method | |
| CN106146449A (en) | The preparation of isorhamnetin and application process thereof in sea-buckthorn pomace | |
| CN110272459A (en) | Two kinds of noval chemical compounds and its antioxidant activity position in root of Paeonia sinjiangensis | |
| CN106769964B (en) | The method of quality control of Herba Achilleae extract | |
| CN105334273B (en) | Detection method of anisetree bark | |
| CN101871924B (en) | Method for synchronous detection of xanthohumol, isoxanthohumol and 8-isopentenylnaringenin in lupulus | |
| CN101766664A (en) | Extraction method of total saponin of Radix Ilicis Asprellae and quality detection method thereof | |
| CN102670698B (en) | The application of Radix Flemingiae Philippinensis extract in preparation control diabetes medicament | |
| CN101176755B (en) | Application of meletin-7-0-glycoside in mass control of cudrania tricuspidata or preparations thereof | |
| CN102735787B (en) | Novel digestion promoting tablet and its relative preparation quality detection method | |
| CN101810650A (en) | Method for measuring content of quercetin p-coumaric acyl glucose rhamnoside contained in folium ginkgo or related preparation thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |