CN1341851A - Quick testing method of rape thio-glycoside and its test board - Google Patents
Quick testing method of rape thio-glycoside and its test board Download PDFInfo
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- CN1341851A CN1341851A CN 01133507 CN01133507A CN1341851A CN 1341851 A CN1341851 A CN 1341851A CN 01133507 CN01133507 CN 01133507 CN 01133507 A CN01133507 A CN 01133507A CN 1341851 A CN1341851 A CN 1341851A
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- sulphur glucoside
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- glucosinolate content
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Abstract
The present invention relates to a quick determination method of rape-glucosinolate and its test plate, and its technical scheme includes the following steps: using 1-5g or rapeseed sample, pulverizing, using 0.20-0.80g of rapeseed powder and placing it into test tube with plug, adding 1.0-5.0 ml of distilled water, uniformly shaking to produce glucosinolate degradation reaction, after a few minuts filtering, taking 20-80 microlitres of filtrate and placing it on the test plate, adding 60-240 microlitres of potassium dihydrogen phosphate aqueous solution to make reaction, after a few minuts forming colour product, its colour is related to glucosinolate content in sample, adopting spectrophotography to measure glucosinolate content by using measurement instrument according to standard curve. Said invented method is simple and convenient, quick in test speed and low in cost.
Description
Technical field
The present invention relates to a kind of being used for carries out the method that speed surveys and measures used test board the rapeseed glucosinolate content, and its application can effectively promote the popularization and the development of double-low rapeseed, to improve the interior quality of rape.
Background technology
Rape is the first oil crops of China, and rapeseed oil accounts for 60% of China's edible vegetable oil total amount, and dregs of rapeseed cake then is the high-quality protein source of feed industry.Double-low rapeseed is a low erucic acid low sulfatide rape, and it has clear and definite limiting the quantity of to erucic acid in the rapeseed and two chemical index of quality in inherence of sulphur glucoside, and wherein the rapeseed oil content of erucic acid should be lower than 2%, and such rapeseed oil is of high nutritive value, and is of value to health; The content of sulphur glucoside should be lower than 30 μ mol/g in the dregs of rapeseed cake, and such grouts toxicity is low, the protein content height, and the amino acid Compositional balance is the forage protein source of high-quality.And the content of its erucic acid and sulphur glucoside can cause common rapeseed oil content of erucic acid height respectively up to more than the 40% and 100 μ mol/g in the common rape, is of low nutritive value, and grouts are difficult to as feed livestock and poultry are poisonous because of sulphur glucoside catabolite.Therefore double-low rapeseed is the direction of present oil plant development.Yet, develop double-low rapeseed, can the mensuration that fast and effeciently carry out the erucic acid glucosinolate content be supplementary measures of being badly in need of solution, otherwise just double-low rapeseed and common rape can't be separated purchase, processing separately, is difficult to realize the excellent usefulness of high-quality of double-low rapeseed.Mensuration with regard to the sulphur glucoside, had at present the liquid chromatogram measuring method of sulphur glucoside, but this method is the program complexity not only, length consuming time, the expense height, and need carry out by the instrument and equipment by costliness in the laboratory, this obviously is unsuitable in the on-the-spot use of plantation wide, that detection limit is big and purchase that distributes; Also have the palladium bichloride method and measure the method for sulphur glucoside, though method of testing is comparatively easy,, instability big because of measuring error can not be practical.
Summary of the invention
To be solved by this invention is the problem that exists at prior art and a kind of simple and effective rape thio-glycoside fast determining method and test board thereof are provided, to promote the popularization and the development of double-low rapeseed.
The technical scheme of rape thio-glycoside fast determining method of the present invention is: after getting the pulverizing of 1-5g vegetable seed sample, take by weighing 0.20-0.80g and place tool plug test tube, add 1.0-5.0ml distilled water, shake up, sulphur glucoside degradation reaction takes place, filter behind the stoichiometric number minute, get 20-80 μ l filtrate and place sulphur glucoside test board, after adding potassium dihydrogen phosphate aqueous solution 60-240 μ l stoichiometric number minute, form coloured product, because the solution colour depth is relevant with glucosinolate content in the sample, adopts spectrophotometric method and can measure glucosinolate content with measuring instrument according to typical curve.
In the such scheme, the fineness after the vegetable seed sample is pulverized is the 30-50 order; The concentration of potassium dihydrogen phosphate aqueous solution is 20-80mmol/L, and preparation method is: fixed molten after getting analysis pure phosphoric acid potassium dihydrogen 6.80g dissolving under the room temperature to 1000ml, behind the ultraviolet disinfection, with a bottle packing; The test result of sulphur glucoside is represented with μ mol/g.
Sulphur glucoside test board of the present invention is made by following method: take by weighing 6.0-6.5mg developer (Chrom G) with deciding molten 10ml behind the anhydrous alcohol solution, get 5-20 μ l and place the every hole of test board, stand-by after vacuum drying 1-2 hour, after taking by weighing the dissolving of 18.24-34.20g dipotassium hydrogen phosphate, fixed molten 1000ml, with standby behind the phosphoric acid accent pH6.0, (Gln MEN) is dissolved in above-mentioned dipotassium hydrogen phosphate damping fluid with sulphur glucoside tested enzyme, compound concentration is 80-180U/ml, get 10-50 μ l and place the every hole of above-mentioned test board, vacuum freezedrying is after 2 hours.
The mechanism of mensuration sulphur glucoside of the present invention is to utilize the selectivity of enzyme, make sulphur glucoside and endogenous enzymes and specificity exogenous enzymes generation reaction in the vegetable seed, the reaction back generates product is formed with characteristic absorption peak again with the developer reaction coloured product, because the solution colour depth is relevant with glucosinolate content in the sample, adopt spectrophotometric method can measure the content of sulphur glucoside with measuring instrument according to typical curve with certain wavelength monochromatic light.
The invention has the advantages that: 1, method of testing is easy, and test speed is fast, can be suitable for the test of various rapeseed kinds; 2, testing expense is low, and the price of tacheometer is low, and volume is little, helps the penetration and promotion of this technology; 3, the mensuration precision is higher, and error is ± 4 μ mol/g, has reached international standard; 4, strong to environmental suitability, be convenient to use at the scene, practical, can effectively promote the industrialized development of double-low rapeseed.
Description of drawings
Fig. 1 is the FB(flow block) of fast determining method of the present invention.
The specific embodiment
Below describe embodiments of the invention in detail.
Embodiment 1
Take by weighing the two No. 3 rape vegetable seed samples of 2g China and pulverize, fineness 40 orders are got 0.50g and are tried in the tool plug Pipe adds 3.0ml distilled water and shakes up generation sulphur glucoside degradation reaction, reaction 3-10 branch under the room temperature Filter behind the clock, get 50 μ l filtrates and place sulphur glucoside test board, add potassium dihydrogen phosphate aqueous solution 150 μ l, React and formed coloured product in 5-15 minute, according to glucosinolate content and form dependency relation between coloured product, Measuring glucosinolate content with measuring instrument is 27.5 μ mol/g.
Embodiment 2
Get two No. 5 rape vegetable seed samples pulverizing among the 4g, fineness 50 orders are got 0.60g in tool plug test tube, Adding 3.6ml distilled water also shakes up, and sulphur glucoside degradation reaction takes place, and reaction was filtered after 8 minutes under the room temperature, Get 60 μ l filtrates and place sulphur glucoside test board, add potassium dihydrogen phosphate aqueous solution 180 μ l, reacted 9 fens The coloured product of bell one-tenth according to dependency relation between glucosinolate content and the coloured product of formation, is surveyed with measuring instrument Deciding glucosinolate content is 25.6 μ mol/g.
Embodiment 3
Embodiment for sulphur glucoside test board of the present invention: take by weighing 6.2mg developer (Chrom G) with not having Fixed molten 10ml got 10 μ l and places the every hole of test board, after vacuum drying 1-2 hour after the water-ethanol dissolving Stand-by, take by weighing 22.8g dipotassium hydrogen phosphate dissolving after, fixed molten 1000ml transfers pH6.0 reserve with phosphoric acid With, sulphur glucoside tested enzyme (Gln MEN) is dissolved in above-mentioned dipotassium hydrogen phosphate buffer solution, compound concentration is 100U/ml gets 20 μ l and places the every hole of above-mentioned test board, and vacuum freezedrying is after 2 hours.
Claims (6)
1, a kind of fast determining method of rape thio-glycoside, after it is characterized in that getting the pulverizing of 1-5g vegetable seed sample, take by weighing 0.20-0.80g and place tool plug test tube, add 1.0-5.0ml distilled water, shake up, sulphur glucoside degradation reaction takes place, filter behind the stoichiometric number minute, get 20-80 μ l filtrate and place sulphur glucoside test board, add potassium dihydrogen phosphate aqueous solution 60-240 μ l stoichiometric number minute after, form coloured product, adopt spectrophotometric method and can measure glucosinolate content with measuring instrument according to typical curve.
2,, it is characterized in that the fineness after the vegetable seed sample is pulverized is the 30-50 order by the fast determining method of the described rape thio-glycoside of claim 1.
3, by the fast determining method of claim 1 or 2 described rape thio-glycosides, the concentration that it is characterized in that potassium dihydrogen phosphate aqueous solution is 20-80mmol/L.
4, press the fast determining method of the described rape erucic acid of claim 3, it is characterized in that taking by weighing 2g vegetable seed sample and pulverize, fineness 40 orders are got 0.50g in tool plug test tube, adding 3.0ml distilled water also shakes up, sulphur glucoside degradation reaction takes place, and reaction was filtered after 3-10 minute under the room temperature, got 50 μ l filtrates and placed sulphur glucoside test board, add potassium dihydrogen phosphate aqueous solution 150 μ l, react and formed coloured product in 5-15 minute, according to glucosinolate content and form correlationship between coloured product, measure glucosinolate content with measuring instrument.
5, press the fast determining method of the described rape erucic acid of claim 3, it is characterized in that getting 4g vegetable seed sample and pulverize, fineness 50 orders are got 0.60g in tool plug test tube, adding 3.6ml distilled water also shakes up, sulphur glucoside degradation reaction takes place, and reaction was filtered after 8 minutes under the room temperature, got 60 μ l filtrates and placed sulphur glucoside test board, add potassium dihydrogen phosphate aqueous solution 180 μ l, react and formed coloured product in 9 minutes, according to glucosinolate content and form correlationship between coloured product, measure glucosinolate content with measuring instrument.
6, a kind of speed of rape thio-glycoside is surveyed test board, it is characterized in that it is made by following method: take by weighing 6.0-6.5mg developer (Chrom G) with deciding molten 10ml behind the anhydrous alcohol solution, get 5-20 μ l and place the every hole of test board, stand-by after vacuum drying 1-2 hour, after taking by weighing the dissolving of 18.24-34.20g dipotassium hydrogen phosphate, fixed molten 1000ml, with standby behind the phosphoric acid accent pH6.0, sulphur glucoside tested enzyme (GlnMEN) is dissolved in above-mentioned dipotassium hydrogen phosphate damping fluid, compound concentration is 80-180U/ml, get 10-50 μ l and place the every hole of above-mentioned test board, vacuum freezedrying is after 2 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01133507 CN1120362C (en) | 2001-09-28 | 2001-09-28 | Quick testing method of rape thio-glycoside and its test board |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01133507 CN1120362C (en) | 2001-09-28 | 2001-09-28 | Quick testing method of rape thio-glycoside and its test board |
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| Publication Number | Publication Date |
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| CN1341851A true CN1341851A (en) | 2002-03-27 |
| CN1120362C CN1120362C (en) | 2003-09-03 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 01133507 Expired - Fee Related CN1120362C (en) | 2001-09-28 | 2001-09-28 | Quick testing method of rape thio-glycoside and its test board |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101832925A (en) * | 2010-05-18 | 2010-09-15 | 浙江大学 | Method and system for quickly detecting content of erucic acid in rapeseed |
| CN101858862A (en) * | 2010-05-18 | 2010-10-13 | 浙江大学 | Method and system for quickly detecting thioglycoside content of rape seeds |
| CN102692387A (en) * | 2012-06-15 | 2012-09-26 | 南京农业大学 | Method for determining total content of glucosinolates in plants |
| EP2520176A1 (en) | 2011-05-02 | 2012-11-07 | ROP Repce-olaj-pellet Kft. | Method for increasing the nutritional value of rapeseed and rape products with decreased oil content |
| CN106644993A (en) * | 2015-10-28 | 2017-05-10 | 广州中大南沙科技创新产业园有限公司 | Spectrophotometer quantitative method of maca glucosinolate |
-
2001
- 2001-09-28 CN CN 01133507 patent/CN1120362C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101832925A (en) * | 2010-05-18 | 2010-09-15 | 浙江大学 | Method and system for quickly detecting content of erucic acid in rapeseed |
| CN101858862A (en) * | 2010-05-18 | 2010-10-13 | 浙江大学 | Method and system for quickly detecting thioglycoside content of rape seeds |
| CN101832925B (en) * | 2010-05-18 | 2011-05-11 | 浙江大学 | Method and system for quickly detecting content of erucic acid in rapeseed |
| EP2520176A1 (en) | 2011-05-02 | 2012-11-07 | ROP Repce-olaj-pellet Kft. | Method for increasing the nutritional value of rapeseed and rape products with decreased oil content |
| CN102692387A (en) * | 2012-06-15 | 2012-09-26 | 南京农业大学 | Method for determining total content of glucosinolates in plants |
| CN106644993A (en) * | 2015-10-28 | 2017-05-10 | 广州中大南沙科技创新产业园有限公司 | Spectrophotometer quantitative method of maca glucosinolate |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1120362C (en) | 2003-09-03 |
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