CN101762575A - Method for identifying authenticity and determining content of lycium barbarum polysaccharides - Google Patents
Method for identifying authenticity and determining content of lycium barbarum polysaccharides Download PDFInfo
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- CN101762575A CN101762575A CN200910265091A CN200910265091A CN101762575A CN 101762575 A CN101762575 A CN 101762575A CN 200910265091 A CN200910265091 A CN 200910265091A CN 200910265091 A CN200910265091 A CN 200910265091A CN 101762575 A CN101762575 A CN 101762575A
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Abstract
The invention relates to a food detection method, in particular to a method for identifying authenticity and determining content of lycium barbarum polysaccharides. The method comprises the following steps: 1) identifying the authenticity; 2) removing monosaccharides, oligosaccharides and glycosides disrupting chemicals and preparing sample solutions; 3) making standard curves; 4) determining polysaccharide content in samples; and 5) calculating result. The invention has the advantages that whether saccharan disrupting chemicals are mixed or not can be identified quickly, and the monosaccharides, the oligosaccharides and the glycosides disrupting chemicals can be effectively removed so as to make the content determination of the lycium barbarum polysaccharides reliable, true and accurate. The invention is suitable for identifying the authenticity of the lycium barbarum polysaccharides mixed with saccharan disrupting chemicals, and is suitable for determining the content of the lycium barbarum polysaccharides with or without monosaccharides, oligosaccharides, glycosides and other disrupting constituents.
Description
Technical field
The present invention relates to a kind of food detection method, specifically relate to a kind of real and fake discrimination and content assaying method thereof of LBP-X.
Background technology
LBP-X is an active substance main in the fruit of Chinese wolfberry, and is water-soluble good because of its content in the fruit of Chinese wolfberry low (3-5%), again not by resin adsorption, so the enrichment of LBP-X and refining relatively more difficult on the commercial production.At present, the content that is provided on the market mostly is the puppet product that added the polysaccharide chaff interference greater than 40% product, and these pseudo-product can't distinguish truth from false with analysis means (colourimetry) of using always at present at all.The existence of pseudo-product brings very big inconvenience and puzzlement for user, supplier and consumer.And being exclusively used in LBP-X real and fake discrimination and Determination on content method thereof, Shang Weijian puts down in writing.
Summary of the invention
The purpose of this invention is to provide a kind of LBP-X real and fake discrimination and content assaying method thereof.
A kind of LBP-X real and fake discrimination of the present invention and content assaying method thereof are achieved by following technical proposals: the real and fake discrimination and the content assaying method thereof of a kind of LBP-X of the present invention comprise the steps:
1). real and fake discrimination
Proterties and sense organ are differentiated
LBP-X should be light yellowish brown to the yellowish-brown uniform powder, has the distinctive fragrance of LBP-X, and solubleness is good in water, and solution is light yellowish brown, and smell, color and solubleness are not inconsistent equal decidable and are pseudo-product;
The polysaccharide chaff interference is differentiated
Sample thief is an amount of, puts in the glass test tube, adds an amount of distilled water normal-temperature dissolution, adds 2-3 and drips indentifying substance KP-1, shakes up, and the observing response phenomenon is positive, and color is deepened rapidly, then contains the polysaccharide chaff interference in the interpret sample;
2). the removal of monose, compound sugar and glycoside chaff interference and sample solution configuration
Get the about 0.4g of LBP-X, the accurate title, decide, be accurate to 0.0001g, place round-bottomed flask, add 80% ethanolic solution 200ml, reflux 1 hour, filtered while hot, residue washs (about 10ml * 10) with hot 80% ethanolic solution gradation, and residue is put in the flask together with filter paper, adds 100ml distilled water, heating was extracted 1 hour, filtered while hot, residue fully washs (about 10ml * 10), merging filtrate and washing lotion with hot distilled water, put cold, move into the 250ml volumetric flask, water is diluted to scale, shakes up promptly to get sample solution;
3). the preparation of typical curve
Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g that are dried to constant weight, puts in the 1000ml volumetric flask, adds the suitable quantity of water dissolving, is diluted to scale, shakes up, and promptly gets reference substance solution;
Precision is measured reference substance solution 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml 1.0ml puts respectively in the tool plug test tube, adding distil water is to 2.0ml respectively, each accurate phenol solution 1.0ml that adds shakes up, the accurate rapidly sulfuric acid 5.0ml that adds, shake up, placed 5 minutes, put in the boiling water bath and heated 15 minutes, being cooled to room temperature after the taking-up rapidly, is blank with the reagent corresponding, measures absorbance at 490nm wavelength place, with the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve;
4). determination of polysaccharide in the sample
Precision is measured sample solution a certain amount of (deciding on sample solution content), puts in the tool plug test tube, and adding distil water is to 2.0ml, method under the preparation of sighting target directrix curve, from " each accurate phenol solution 1.0ml that adds ", measure absorbance, find glucose content in the colour developing liquid according to typical curve;
5). the result calculates
Computing formula
W in the formula---polyoses content (%)
P---glucose content (g) in the colour developing liquid
F---the conversion factor of 3.19 glucose conversion polysaccharide
The quality of m---sample (g)
V---draw the volume (ml) of sample solution.
A kind of LBP-X real and fake discrimination of the present invention and content assaying method thereof have following beneficial effect compared with prior art: the inventive method is in conjunction with the peculiar physicochemical property of LBP-X, by its real and fake discrimination and assay thereof, sample is dissolved in (normal temperature) in the distilled water, add indentifying substance KP-1, distinguish whether be mixed with the polysaccharide chaff interference.The sample that does not contain chaff interference, use 80% alcohol reflux, after removing interfering materials such as monose compound sugar and glycoside, the polysaccharide composition is under effect of sulfuric acid, be hydrolyzed into monose earlier, and dehydration generates the alditol derivant rapidly, is condensed into colored compound with phenol then, measures polyoses content in suitable wavelength place with spectrophotometric method.The present invention can effectively differentiate whether be mixed with the polysaccharide chaff interference fast, and can effectively remove monose, compound sugar and glycoside interference component, makes the LBP-X assay reliable, true, accurate.The inventive method is applicable to the LBP-X real and fake discrimination that is mixed with the polysaccharide chaff interference.Be applicable to the assay that contains or do not contain the LBP-X of interference components such as monose, compound sugar and glycoside.
Embodiment
Below in conjunction with embodiment a kind of LBP-X real and fake discrimination of the present invention and content assaying method technical scheme thereof are further described.
The real and fake discrimination and the content assaying method thereof of a kind of LBP-X of the present invention comprise the steps:
1). real and fake discrimination
Proterties and sense organ are differentiated
LBP-X should be light yellowish brown to the yellowish-brown uniform powder, has the distinctive fragrance of LBP-X, and solubleness is good in water, and solution is light yellowish brown, and smell, color and solubleness are not inconsistent equal decidable and are pseudo-product;
The polysaccharide chaff interference is differentiated
Sample thief is an amount of, puts in the glass test tube, adds an amount of distilled water normal-temperature dissolution, adds 2-3 and drips indentifying substance KP-1, shakes up, and the observing response phenomenon is positive, and color is deepened rapidly, then contains the polysaccharide chaff interference in the interpret sample;
2). the removal of monose, compound sugar and glycoside chaff interference and sample solution configuration
Get the about 0.4g of LBP-X, the accurate title, decide, be accurate to 0.0001g, place round-bottomed flask, add 80% ethanolic solution 200ml, reflux 1 hour, filtered while hot, residue washs (about 10ml * 10) with hot 80% ethanolic solution gradation, and residue is put in the flask together with filter paper, adds 100ml distilled water, heating was extracted 1 hour, filtered while hot, residue fully washs (about 10ml * 10), merging filtrate and washing lotion with hot distilled water, put cold, move into the 250ml volumetric flask, water is diluted to scale, shakes up promptly to get sample solution;
3). the preparation of typical curve
Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g that are dried to constant weight, puts in the 1000ml volumetric flask, adds the suitable quantity of water dissolving, is diluted to scale, shakes up, and promptly gets reference substance solution;
Precision is measured reference substance solution 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml 1.0ml puts respectively in the tool plug test tube, adding distil water is to 2.0ml respectively, each accurate phenol solution 1.0ml that adds shakes up, the accurate rapidly sulfuric acid 5.0ml that adds, shake up, placed 5 minutes, put in the boiling water bath and heated 15 minutes, being cooled to room temperature after the taking-up rapidly, is blank with the reagent corresponding, measures absorbance at 490nm wavelength place, with the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve;
4). determination of polysaccharide in the sample
Precision is measured sample solution a certain amount of (deciding on sample solution content), puts in the tool plug test tube, and adding distil water is to 2.0ml, method under the preparation of sighting target directrix curve, from " each accurate phenol solution 1.0ml that adds ", measure absorbance, find glucose content in the colour developing liquid according to typical curve;
5). the result calculates
Computing formula
W in the formula---polyoses content (%)
P---glucose content (g) in the colour developing liquid
F---the conversion factor of 3.19 glucose conversion polysaccharide
The quality of m---sample (g)
V---draw the volume (ml) of sample solution.
Described real and fake discrimination and assay thereof have repeatability, and each sample is got two parallel samples and measured, and are measurement result with its arithmetic mean, and allowing relative deviation is 5%.
Get the about 0.4g of LBP-X in the removal of described monose, compound sugar and glycoside chaff interference and the sample solution configuration step, the accurate title, decide, and is accurate to 0.0001g; Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g that are dried to constant weight in the preparation process of described typical curve, is accurate to 0.0001g.
The used reagent of described discriminating and assay thereof is:
95% ethanol or absolute ethyl alcohol (analyzing pure), indentifying substance KP-1, sulfuric acid (analyzing pure)
Phenol liquid: get phenol 100g, with aluminium flake 0.1g and sodium bicarbonate 0.05g, 172 ℃ of cuts are collected in distillation, take by weighing this cut 10g, add water 150ml, place brown bottle promptly.
The used instrument of described method is:
Analytical balance
The glass reflux
Ultraviolet-visible pectrophotometer
The electric heating constant temperature water-bath
The glass distilling apparatus
250ml volumetric flask, transfer pipet, tool plug test tube, beaker, glass rod.
Embodiment 1.
The present invention is applicable to the LBP-X real and fake discrimination that is mixed with the polysaccharide chaff interference.
Be applicable to the assay that contains or do not contain the LBP-X of interference components such as monose, compound sugar and glycoside.
1. reference document
Clause in the following files becomes the clause of this standard by quotation of this standard.Every reference document with bearing date, its all subsequently modification single (not comprising the content of correcting errors in printing) or revision all are not suitable for this standard, however whether each side's research that encouragement is reached an agreement according to this standard can use the latest edition of these files.Every undated reference file, its latest edition is fit to this standard.
GB/T 18672-2002 appendix A LBP-X is measured
2. principle summary
Sample is dissolved in (normal temperature) in the distilled water, adds indentifying substance KP-1, distinguish whether be mixed with the polysaccharide chaff interference.The sample that does not contain chaff interference, use 80% alcohol reflux, after removing interfering materials such as monose compound sugar and glycoside, the polysaccharide composition is under effect of sulfuric acid, be hydrolyzed into monose earlier, and dehydration generates the alditol derivant rapidly, is condensed into colored compound with phenol then, measures polyoses content in suitable wavelength place with spectrophotometric method.
3. main agents and instrument
3.1. main agents
95% ethanol or absolute ethyl alcohol (analyzing pure), indentifying substance KP-1, sulfuric acid (analyzing pure)
Phenol liquid: get phenol 100g, with aluminium flake 0.1g and sodium bicarbonate 0.05g, 172 ℃ of cuts are collected in distillation, take by weighing this cut 10g, add water 150ml, place brown bottle promptly.
3.2. instrument
Analytical balance
The glass reflux
Ultraviolet-visible pectrophotometer
The electric heating constant temperature water-bath
The glass distilling apparatus
250ml volumetric flask, transfer pipet, tool plug test tube, beaker, glass rod
4. real and fake discrimination and assay
4.1. real and fake discrimination
4.1.1. proterties and sense organ are differentiated
LBP-X should be light yellowish brown to the yellowish-brown uniform powder, has the distinctive fragrance of LBP-X, and solubleness is good in water, and solution is light yellowish brown, and smell, color and solubleness are not inconsistent equal decidable and are pseudo-product.
4.1.2. the polysaccharide chaff interference is differentiated
Sample thief is an amount of, puts in the glass test tube, adds an amount of distilled water (normal temperature) dissolving, adds 2-3 and drips indentifying substance KP-1, shake up, and the observing response phenomenon, be positive (color is deepened rapidly) then contains the polysaccharide chaff interference in the interpret sample.
4.2. the removal of monose, compound sugar and glycoside chaff interference and sample solution configuration
Get the about 0.4g of LBP-X, the accurate title, decide, be accurate to 0.0001g, place round-bottomed flask, add 80% ethanolic solution 200ml, reflux 1 hour, filtered while hot, residue washs (about 10ml * 10) with hot 80% ethanolic solution gradation, and residue is put in the flask together with filter paper, adds 100ml distilled water, heating was extracted 1 hour, filtered while hot, residue fully washs (about 10ml * 10), merging filtrate and washing lotion with hot distilled water, put cold, move into the 250ml volumetric flask, water is diluted to scale, shakes up promptly to get sample solution.
4.3. the preparation of typical curve
Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g (being accurate to 0.0001g) that are dried to constant weight, puts in the 1000ml volumetric flask, adds the suitable quantity of water dissolving, is diluted to scale, shakes up, and promptly gets reference substance solution.
Precision is measured reference substance solution 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml 1.0ml puts respectively in the tool plug test tube, adding distil water is to 2.0ml respectively, each accurate phenol solution 1.0ml that adds shakes up, the accurate rapidly sulfuric acid 5.0ml that adds, shake up, placed 5 minutes, put in the boiling water bath and heated 15 minutes, being cooled to room temperature after the taking-up rapidly, is blank with the reagent corresponding, measures absorbance at 490nm wavelength place, with the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve.
4.4. determination of polysaccharide in the sample
Precision is measured sample solution a certain amount of (deciding on sample solution content), puts in the tool plug test tube, and adding distil water is to 2.0ml, method under the preparation of sighting target directrix curve, from " each accurate phenol solution 1.0ml that adds ", measure absorbance, find glucose content in the colour developing liquid according to typical curve.
4.5. the result calculates
Computing formula
W in the formula---polyoses content (%)
P---glucose content (g) in the colour developing liquid
F---the conversion factor of 3.19 glucose conversion polysaccharide
The quality of m---sample (g)
V---draw the volume (ml) of sample solution
4.6 repeatability
Each sample is got two parallel samples and is measured, and is measurement result with its arithmetic mean, and allowing relative deviation is 5%.
Claims (5)
1. the real and fake discrimination of a LBP-X and content assaying method thereof, its feature comprises the steps: in described real and fake discrimination and content assaying method thereof
1). real and fake discrimination
Proterties and sense organ are differentiated
LBP-X should be light yellowish brown to the yellowish-brown uniform powder, has the distinctive fragrance of LBP-X, and solubleness is good in water, and solution is light yellowish brown, and smell, color and solubleness are not inconsistent equal decidable and are pseudo-product;
The polysaccharide chaff interference is differentiated
Sample thief is an amount of, puts in the glass test tube, adds an amount of distilled water normal-temperature dissolution, adds 2-3 and drips indentifying substance KP-1, shakes up, and the observing response phenomenon is positive, and color is deepened rapidly, then contains the polysaccharide chaff interference in the interpret sample;
2). the removal of monose, compound sugar and glycoside chaff interference and sample solution configuration
Get the about 0.4g of LBP-X, the accurate title, decide, be accurate to 0.000lg, place round-bottomed flask, add 80% ethanolic solution 200ml, reflux 1 hour, filtered while hot, residue washs (about 10ml * 10) with hot 80% ethanolic solution gradation, and residue is put in the flask together with filter paper, adds 100ml distilled water, heating was extracted 1 hour, filtered while hot, residue fully washs (about 10ml * 10), merging filtrate and washing lotion with hot distilled water, put cold, move into the 250ml volumetric flask, water is diluted to scale, shakes up promptly to get sample solution;
3). the preparation of typical curve
Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g that are dried to constant weight, puts in the 1000ml volumetric flask, adds the suitable quantity of water dissolving, is diluted to scale, shakes up, and promptly gets reference substance solution;
Precision is measured reference substance solution 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml 1.0ml puts respectively in the tool plug test tube, adding distil water is to 2.0ml respectively, each accurate phenol solution 1.0ml that adds shakes up, the accurate rapidly sulfuric acid 5.0ml that adds, shake up, placed 5 minutes, put in the boiling water bath and heated 15 minutes, being cooled to room temperature after the taking-up rapidly, is blank with the reagent corresponding, measures absorbance at 490nm wavelength place, with the absorbance is ordinate, and concentration is horizontal ordinate, the drawing standard curve;
4). determination of polysaccharide in the sample
Precision is measured sample solution a certain amount of (deciding on sample solution content), puts in the tool plug test tube, and adding distil water is to 2.0ml, method under the preparation of sighting target directrix curve, from " each accurate phenol solution 1.0ml that adds ", measure absorbance, find glucose content in the colour developing liquid according to typical curve;
5). the result calculates
Computing formula
W in the formula---polyoses content (%)
P---glucose content (g) in the colour developing liquid
F---the conversion factor of 3.19 glucose conversion polysaccharide
The quality of m---sample (g)
V---draw the volume (ml) of sample solution.
2. the real and fake discrimination of LBP-X according to claim 1 and content assaying method thereof, it is characterized in that: described real and fake discrimination and assay thereof have repeatability, each sample is got two parallel samples and is measured, and is measurement result with its arithmetic mean, and allowing relative deviation is 5%.
3. the real and fake discrimination of LBP-X according to claim 1 and content assaying method thereof, it is characterized in that: get the about 0.4g of LBP-X in the removal of described monose, compound sugar and glycoside chaff interference and the sample solution configuration step, the accurate title, decide, and is accurate to 0.0001g; Precision takes by weighing 105 ℃ of anhydrous dextrose reference substance 0.1g that are dried to constant weight in the preparation process of described typical curve, is accurate to 0.0001g.
4. the real and fake discrimination of LBP-X according to claim 1 and content assaying method thereof is characterized in that, the used reagent of described discriminating and assay thereof is:
95% ethanol or absolute ethyl alcohol (analyzing pure), indentifying substance KP-1, sulfuric acid (analyzing pure)
Phenol liquid: get phenol 100g, with aluminium flake 0.1g and sodium bicarbonate 0.05g, 172 ℃ of cuts are collected in distillation, take by weighing this cut 10g, add water 150ml, place brown bottle promptly.
5. the real and fake discrimination of LBP-X according to claim 1 and content assaying method thereof is characterized in that, the used instrument of described method is:
Analytical balance
The glass reflux
Ultraviolet-visible pectrophotometer
The electric heating constant temperature water-bath
The glass distilling apparatus
250ml volumetric flask, transfer pipet, tool plug test tube, beaker, glass rod.
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| CN200910265091A CN101762575A (en) | 2009-12-28 | 2009-12-28 | Method for identifying authenticity and determining content of lycium barbarum polysaccharides |
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| CN200910265091A CN101762575A (en) | 2009-12-28 | 2009-12-28 | Method for identifying authenticity and determining content of lycium barbarum polysaccharides |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102495167A (en) * | 2011-12-09 | 2012-06-13 | 劲牌有限公司 | Method for detecting lycium barbarum polysaccharide in lycium barbarum polysaccharide extract |
| CN102879347A (en) * | 2012-10-18 | 2013-01-16 | 中国科学院西北高原生物研究所 | Detection method of polysaccharide content in medlar polysaccharide extract doped with saccharide chaff interferent |
| CN103901031A (en) * | 2014-04-14 | 2014-07-02 | 中国农业科学院兰州畜牧与兽药研究所 | Method for rapid and high-flux determination of polysaccharide content based on sulfuric acid and phenol |
| CN104406978A (en) * | 2014-12-31 | 2015-03-11 | 青海康普生物科技股份有限公司 | Simple method of true and false identification of wolfberry polysaccharides |
| CN109374776A (en) * | 2018-12-12 | 2019-02-22 | 中国科学院兰州化学物理研究所 | Identify the method for polysaccharides quality using liquid chromatogram |
-
2009
- 2009-12-28 CN CN200910265091A patent/CN101762575A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102495167A (en) * | 2011-12-09 | 2012-06-13 | 劲牌有限公司 | Method for detecting lycium barbarum polysaccharide in lycium barbarum polysaccharide extract |
| CN102879347A (en) * | 2012-10-18 | 2013-01-16 | 中国科学院西北高原生物研究所 | Detection method of polysaccharide content in medlar polysaccharide extract doped with saccharide chaff interferent |
| CN102879347B (en) * | 2012-10-18 | 2016-01-20 | 中国科学院西北高原生物研究所 | Be mixed with the detection method of polyoses content in the Lycium barbarum polysaccharide extract of carbohydrate chaff interference |
| CN103901031A (en) * | 2014-04-14 | 2014-07-02 | 中国农业科学院兰州畜牧与兽药研究所 | Method for rapid and high-flux determination of polysaccharide content based on sulfuric acid and phenol |
| CN104406978A (en) * | 2014-12-31 | 2015-03-11 | 青海康普生物科技股份有限公司 | Simple method of true and false identification of wolfberry polysaccharides |
| CN109374776A (en) * | 2018-12-12 | 2019-02-22 | 中国科学院兰州化学物理研究所 | Identify the method for polysaccharides quality using liquid chromatogram |
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Open date: 20100630 |