CN104949933A - Detection method for Korean pine kernel polysaccharide - Google Patents
Detection method for Korean pine kernel polysaccharide Download PDFInfo
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- CN104949933A CN104949933A CN201510365851.0A CN201510365851A CN104949933A CN 104949933 A CN104949933 A CN 104949933A CN 201510365851 A CN201510365851 A CN 201510365851A CN 104949933 A CN104949933 A CN 104949933A
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- korean pine
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Abstract
The invention discloses a detection method for Korean pine kernel polysaccharide. The method comprises the following steps: adding dry Korean pine kernels into distilled water, performing reflux extraction, performing and frozen centrifuging on an extracting liquid, and performing nanofiltration concentration on a centrifuged supernatant I; adding a mixed liquid of n-amyl alcohol and ethyl acetate into a concentrated liquid, stirring, standing, and removing sediments I; adding a 95% ethanol aqueous solution into a supernatant II, centrifuging, and removing supernatant III; adding a mixed liquid of n-butyl alcohol and trichloromethane into sediments II, stirring, standing, and removing sediments III; adding the 95% ethanol aqueous solution into a supernatant IV, centrifuging, and washing sediments IV with absolute ethyl alcohol so as to obtain the Korean pine kernel polysaccharide; and determining the absorbance value of the Korean pine kernel polysaccharide by adopting an ultraviolet spectrophotometer at a wave length being 490nm, and determining the content of the Korean pine kernel polysaccharide by taking glucose as the contrast. The detection method is simple, the required sample amount is small, and a practical and effective extraction and analysis method can be provided for further research on drug actions of the Korean pine kernel polysaccharide.
Description
Technical field
The invention belongs to food analysis technical field, be specifically related to a kind of detection method of Korean pine kind benevolence polysaccharide.
Background technology
In China, the history of edible Korean pine kind benevolence oneself have more than 3000 year, since the Tang Dynasty, Korean pine kind benevolence just becomes the pot foods that people like, Song dynasty is considered as Korean pine kind benevolence " peanut " of promoting longevity especially, it was both regarded as ambrosia, made again dietotherapy good merchantable brand, extremely the high praise of doctor, nutrition man.Compendium of Material Medica one is recorded: " pine nut taste is sweet, warm in nature, nontoxic.Cure mainly: bone joint rheumatism, dizzy, wines used as antirheumatic, moisturizing viscera, allays one's hunger, and by wandering arthritis cold air, complement is empty, skin care, and clothes are made light of one's life by commiting suicide old for a long time, separately have moistening lung function, control constipation cough ".
Along with the rise pursuing wholefood tide, the exploitation of Korean pine kind benevolence Related product becomes a new highlight in food and health products exploitation gradually, and the carbohydrates total content in Korean pine kind benevolence is 2.2%.Polysaccharide is the natural polymer polymer that a class is formed by connecting by sugar former times key by aldose or ketose, is one of molecular basis forming life." polysaccharide biology " is the new forward position of life science, the biological information amount of polysaccharide than protein and adjust taller go out more than an order of magnitude.
Research finds that polysaccharide compound contained by kindred plant is not a para-immunity correctives, and it has Tumor suppression growth, and immune cell activated, improves the effects such as body's immunity.To not normal cells spinoff.Up to the present, the most important with water-soluble polysaccharide in the polysaccharide compound separated from plant.Shown by the research movable in vivo to immune substance, biological membrane and various bioactivators, carbohydrate function is in vivo not only provide energy and participation structure tissue, also there is diversified biological function simultaneously, important role is played the part of in biological life activity, participate in the various activities of cell, especially function and the application of a polysaccharide focus paying close attention to of people especially in recent years, large quantity research both domestic and external proves, polysaccharide has many-sided biologically active and function.
At present, the research of carrying out extracting the assay of also Optimized Extraction Process and extraction product to Korean pine kind benevolence polysaccharide is also little.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art and the detection method that a kind of Korean pine kind benevolence polysaccharide is provided, this detection method is simple, required sample size is few, for the drug action studying Korean pine kind benevolence polysaccharide further provides a kind of extraction and analytical method of practicability and effectiveness.
The detection method of Korean pine kind benevolence polysaccharide, comprises the following steps:
Step 1, gets dry Korean pine kind benevolence, adds distilled water, refluxing extraction;
Step 2, by step 1 gained extract refrigerated centrifuge, the centrifugal supernatant nanofiltration of gained concentrates;
Step 3, adds the mixed liquor of n-amyl alcohol and ethyl acetate in step 2 gained concentrate, leave standstill, abandon precipitation after stirring;
Step 4, adds 95% ethanol water in step 3 gained supernatant, centrifugal, abandons supernatant;
Step 5, adds the mixed liquor of normal butyl alcohol and methenyl choloride, leaves standstill, abandon precipitation after stirring in step 4 gained precipitation;
Step 6, adds 95% ethanol water in step 5 gained supernatant, centrifugal, and gained precipitation, through absolute ethanol washing, obtains Korean pine kind benevolence polysaccharide;
Step 7, by step 6 gained Korean pine kind benevolence polysaccharide ultraviolet spectrophotometer, 490nm place measures absorbance, contrasts with glucose, measures content.
As the further improvement of foregoing invention, in step 1, the consumption of Korean pine kind benevolence is 20 ~ 60g, and the consumption of distilled water is 10 ~ 50mL.
As the further improvement of foregoing invention, in step 1, refluxing extraction temperature is 70 ~ 90 DEG C, and the time is 1 ~ 3h.
As the further improvement of foregoing invention, in step 2, NF membrane aperture is 100 ~ 200nm.
As the further improvement of foregoing invention, in step 2, the condition of refrigerated centrifuge is-10 DEG C, 3000 ~ 5000rpm, and the time is 10 ~ 15min.
As the further improvement of foregoing invention, in step 3, the consumption of concentrate is 5 ~ 10mL, and the consumption of the mixed liquor of n-amyl alcohol and ethyl acetate is 25 ~ 45 mL, and in mixed liquor, the volume ratio of n-amyl alcohol and ethyl acetate is 1 ~ 5:3 ~ 9.
As the further improvement of foregoing invention, in step 4, the volume ratio of upper cleer and peaceful 95% ethanol water is 2 ~ 9:7 ~ 14.
As the further improvement of foregoing invention, precipitating consumption in step 5 is 5 ~ 20g, and the consumption of the mixed liquor of normal butyl alcohol and methenyl choloride is 30 ~ 50mL, and in mixed liquor, the volume ratio of normal butyl alcohol and methenyl choloride is 1 ~ 6:4 ~ 12.
As the further improvement of foregoing invention, in step 6, the volume ratio of upper cleer and peaceful 95% ethanol water is 3 ~ 12:5 ~ 25.
Detection method of the present invention is simple, and required sample size is few, for the drug action studying Korean pine kind benevolence polysaccharide further provides a kind of extraction and analytical method of practicability and effectiveness.
Embodiment
Embodiment 1
The detection method of Korean pine kind benevolence polysaccharide, comprises the following steps:
Step 1, gets dry Korean pine kind benevolence, adds distilled water, refluxing extraction;
Step 2, by step 1 gained extract refrigerated centrifuge, the centrifugal supernatant nanofiltration of gained concentrates;
Step 3, adds the mixed liquor of n-amyl alcohol and ethyl acetate in step 2 gained concentrate, leave standstill, abandon precipitation after stirring;
Step 4, adds 95% ethanol water in step 3 gained supernatant, centrifugal, abandons supernatant;
Step 5, adds the mixed liquor of normal butyl alcohol and methenyl choloride, leaves standstill, abandon precipitation after stirring in step 4 gained precipitation;
Step 6, adds 95% ethanol water in step 5 gained supernatant, centrifugal, and gained precipitation, through absolute ethanol washing, obtains Korean pine kind benevolence polysaccharide;
Step 7, by step 6 gained Korean pine kind benevolence polysaccharide ultraviolet spectrophotometer, 490nm place measures absorbance, contrasts with glucose, measures content.
In step 1, the consumption of Korean pine kind benevolence is 20g, and the consumption of distilled water is 10mL.
In step 1, refluxing extraction temperature is 70 DEG C, and the time is 1h.
In step 2, NF membrane aperture is 100nm.
In step 2, the condition of refrigerated centrifuge is-10 DEG C, 3000rpm, and the time is 10min.
In step 3, the consumption of concentrate is 5mL, and the consumption of the mixed liquor of n-amyl alcohol and ethyl acetate is 25mL, and in mixed liquor, the volume ratio of n-amyl alcohol and ethyl acetate is 1:3.
In step 4, the volume ratio of upper cleer and peaceful 95% ethanol water is 2:7.
Precipitating consumption in step 5 is 5g, and the consumption of the mixed liquor of normal butyl alcohol and methenyl choloride is 30mL, and in mixed liquor, the volume ratio of normal butyl alcohol and methenyl choloride is 1:4.
In step 6, the volume ratio of upper cleer and peaceful 95% ethanol water is 3:5.
Detect through above-mentioned detection method, Korean pine kind benevolence polyoses content is 58.9%.
Embodiment 2
The detection method of Korean pine kind benevolence polysaccharide, comprises the following steps:
Step 1, gets dry Korean pine kind benevolence, adds distilled water, refluxing extraction;
Step 2, by step 1 gained extract refrigerated centrifuge, the centrifugal supernatant nanofiltration of gained concentrates;
Step 3, adds the mixed liquor of n-amyl alcohol and ethyl acetate in step 2 gained concentrate, leave standstill, abandon precipitation after stirring;
Step 4, adds 95% ethanol water in step 3 gained supernatant, centrifugal, abandons supernatant;
Step 5, adds the mixed liquor of normal butyl alcohol and methenyl choloride, leaves standstill, abandon precipitation after stirring in step 4 gained precipitation;
Step 6, adds 95% ethanol water in step 5 gained supernatant, centrifugal, and gained precipitation, through absolute ethanol washing, obtains Korean pine kind benevolence polysaccharide;
Step 7, by step 6 gained Korean pine kind benevolence polysaccharide ultraviolet spectrophotometer, 490nm place measures absorbance, contrasts with glucose, measures content.
In step 1, the consumption of Korean pine kind benevolence is 45g, and the consumption of distilled water is 28mL.
In step 1, refluxing extraction temperature is 80 DEG C, and the time is 2h.
In step 2, NF membrane aperture is 150nm.
In step 2, the condition of refrigerated centrifuge is-10 DEG C, 4000rpm, and the time is 12min.
In step 3, the consumption of concentrate is 8mL, and the consumption of the mixed liquor of n-amyl alcohol and ethyl acetate is 30mL, and in mixed liquor, the volume ratio of n-amyl alcohol and ethyl acetate is 3:7.
In step 4, the volume ratio of upper cleer and peaceful 95% ethanol water is 6:11.
Precipitating consumption in step 5 is 5g, and the consumption of the mixed liquor of normal butyl alcohol and methenyl choloride is 45mL, and in mixed liquor, the volume ratio of normal butyl alcohol and methenyl choloride is 5:9.
In step 6, the volume ratio of upper cleer and peaceful 95% ethanol water is 7:22.
Detect through above-mentioned detection method, Korean pine kind benevolence polyoses content is 65.4%.
Embodiment 3
The detection method of Korean pine kind benevolence polysaccharide, comprises the following steps:
Step 1, gets dry Korean pine kind benevolence, adds distilled water, refluxing extraction;
Step 2, by step 1 gained extract refrigerated centrifuge, the centrifugal supernatant nanofiltration of gained concentrates;
Step 3, adds the mixed liquor of n-amyl alcohol and ethyl acetate in step 2 gained concentrate, leave standstill, abandon precipitation after stirring;
Step 4, adds 95% ethanol water in step 3 gained supernatant, centrifugal, abandons supernatant;
Step 5, adds the mixed liquor of normal butyl alcohol and methenyl choloride, leaves standstill, abandon precipitation after stirring in step 4 gained precipitation;
Step 6, adds 95% ethanol water in step 5 gained supernatant, centrifugal, and gained precipitation, through absolute ethanol washing, obtains Korean pine kind benevolence polysaccharide;
Step 7, by step 6 gained Korean pine kind benevolence polysaccharide ultraviolet spectrophotometer, 490nm place measures absorbance, contrasts with glucose, measures content.
In step 1, the consumption of Korean pine kind benevolence is 60g, and the consumption of distilled water is 50mL.
In step 1, refluxing extraction temperature is 90 DEG C, and the time is 3h.
In step 2, NF membrane aperture is 200nm.
In step 2, the condition of refrigerated centrifuge is-10 DEG C, 5000rpm, and the time is 15min.
In step 3, the consumption of concentrate is 10mL, and the consumption of the mixed liquor of n-amyl alcohol and ethyl acetate is 45mL, and in mixed liquor, the volume ratio of n-amyl alcohol and ethyl acetate is 5:9.
In step 4, the volume ratio of upper cleer and peaceful 95% ethanol water is 9:14.
Precipitating consumption in step 5 is 20g, and the consumption of the mixed liquor of normal butyl alcohol and methenyl choloride is 50mL, and in mixed liquor, the volume ratio of normal butyl alcohol and methenyl choloride is 6:12.
In step 6, the volume ratio of upper cleer and peaceful 95% ethanol water is 12:25.
Detect through above-mentioned detection method, Korean pine kind benevolence polyoses content is 67.8%.
Detection method provided by the invention is simple, and required sample size is few, for the drug action studying Korean pine kind benevolence polysaccharide further provides a kind of extraction and analytical method of practicability and effectiveness.
Claims (9)
1. the detection method of Korean pine kind benevolence polysaccharide, is characterized in that: comprise the following steps:
Step 1, gets dry Korean pine kind benevolence, adds distilled water, refluxing extraction;
Step 2, by step 1 gained extract refrigerated centrifuge, the centrifugal supernatant nanofiltration of gained concentrates;
Step 3, adds the mixed liquor of n-amyl alcohol and ethyl acetate in step 2 gained concentrate, leave standstill, abandon precipitation after stirring;
Step 4, adds 95% ethanol water in step 3 gained supernatant, centrifugal, abandons supernatant;
Step 5, adds the mixed liquor of normal butyl alcohol and methenyl choloride, leaves standstill, abandon precipitation after stirring in step 4 gained precipitation;
Step 6, adds 95% ethanol water in step 5 gained supernatant, centrifugal, and gained precipitation, through absolute ethanol washing, obtains Korean pine kind benevolence polysaccharide;
Step 7, by step 6 gained Korean pine kind benevolence polysaccharide ultraviolet spectrophotometer, 490nm place measures absorbance, contrasts with glucose, measures content.
2. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, is characterized in that: in step 1, the consumption of Korean pine kind benevolence is 20 ~ 60g, and the consumption of distilled water is 10 ~ 50mL.
3. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, it is characterized in that: in step 1, refluxing extraction temperature is 70 ~ 90 DEG C, the time is 1 ~ 3h.
4. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, is characterized in that: in step 2, NF membrane aperture is 100 ~ 200nm.
5. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, is characterized in that: in step 2, the condition of refrigerated centrifuge is-10 DEG C, 3000 ~ 5000rpm, and the time is 10 ~ 15min.
6. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, it is characterized in that: in step 3, the consumption of concentrate is 5 ~ 10mL, the consumption of the mixed liquor of n-amyl alcohol and ethyl acetate is 25 ~ 45 mL, and in mixed liquor, the volume ratio of n-amyl alcohol and ethyl acetate is 1 ~ 5:3 ~ 9.
7. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, is characterized in that: in step 4, the volume ratio of upper cleer and peaceful 95% ethanol water is 2 ~ 9:7 ~ 14.
8. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, it is characterized in that: precipitating consumption in step 5 is 5 ~ 20g, the consumption of the mixed liquor of normal butyl alcohol and methenyl choloride is 30 ~ 50mL, and in mixed liquor, the volume ratio of normal butyl alcohol and methenyl choloride is 1 ~ 6:4 ~ 12.
9. the detection method of Korean pine kind benevolence polysaccharide according to claim 1, is characterized in that: in step 6, the volume ratio of upper cleer and peaceful 95% ethanol water is 3 ~ 12:5 ~ 25.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5595893A (en) * | 1992-06-19 | 1997-01-21 | Iowa State University Research Foundation, Inc. | Immobilization of microorganisms on a support made of synthetic polymer and plant material |
| CN1317495A (en) * | 2001-04-23 | 2001-10-17 | 南京大学 | Ocean thalassiomycete hypoxylon polyose and its extracting process and application |
| CN1803789A (en) * | 2006-01-20 | 2006-07-19 | 华中科技大学 | Method for extracting glycyrrhizicacid, licorice flavone and licorice polysaccharide |
| CN1836676A (en) * | 2005-03-23 | 2006-09-27 | 吴逸芳 | Method for preparing and controlling the quality of lucid ganoderma spore powder soft capsule |
| CN101116722A (en) * | 2006-08-01 | 2008-02-06 | 天津市轩宏医药技术有限公司 | Pharmaceutical formulations with the raw material containing panax, ophiopogon root and schisandra fruit, processes for their preparation, the raw material and the quality control method for the prepa |
-
2015
- 2015-06-29 CN CN201510365851.0A patent/CN104949933A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5595893A (en) * | 1992-06-19 | 1997-01-21 | Iowa State University Research Foundation, Inc. | Immobilization of microorganisms on a support made of synthetic polymer and plant material |
| CN1317495A (en) * | 2001-04-23 | 2001-10-17 | 南京大学 | Ocean thalassiomycete hypoxylon polyose and its extracting process and application |
| CN1836676A (en) * | 2005-03-23 | 2006-09-27 | 吴逸芳 | Method for preparing and controlling the quality of lucid ganoderma spore powder soft capsule |
| CN1803789A (en) * | 2006-01-20 | 2006-07-19 | 华中科技大学 | Method for extracting glycyrrhizicacid, licorice flavone and licorice polysaccharide |
| CN101116722A (en) * | 2006-08-01 | 2008-02-06 | 天津市轩宏医药技术有限公司 | Pharmaceutical formulations with the raw material containing panax, ophiopogon root and schisandra fruit, processes for their preparation, the raw material and the quality control method for the prepa |
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Application publication date: 20150930 |
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