CN105203532A - Detection method for urea and total ammonia nitrogen in bean sprouts - Google Patents
Detection method for urea and total ammonia nitrogen in bean sprouts Download PDFInfo
- Publication number
- CN105203532A CN105203532A CN201510582321.1A CN201510582321A CN105203532A CN 105203532 A CN105203532 A CN 105203532A CN 201510582321 A CN201510582321 A CN 201510582321A CN 105203532 A CN105203532 A CN 105203532A
- Authority
- CN
- China
- Prior art keywords
- urea
- bean sprouts
- urase
- ammonia nitrogen
- detection method
- 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.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a detection method for urea and total ammonia nitrogen in bean sprouts. The detection method comprises the steps of chopping the bean sprouts, soaking the chopped bean sprouts with water, adding urease into a supernatant so as to catalyze the supernatant, adding a color developing agent to carry out color development, and comparing colors with a color gradation card, thereby obtaining urea content. The use amount ratio of a sample to the urease to the color developing agent is 0.5mL: (20-200)uL: (20-200)uL, and the enzyme activity of the urease is 30-180U/mL. The detection method provided by the invention is low in cost and high in sensitivity and can be used for rapidly, accurately and conveniently detecting urea content greater than or equal to 3mg/L and ammonia nitrogen content greater than or equal to 1.5mg/L.
Description
Technical field
Chemical field of the present invention, particularly relates to urea and total ammonia nitrogen detection method in a kind of bean sprouts.
Background technology
The normal production cycles of bean sprouts mostly is about 11 ~ 15 days, but some retailers are in order to improve bean sprouts output, shortens the production cycle, sprays the chemical fertilizer such as urea, ammonium nitrate, ammonium sulfate and root-free agent and hormone expedites the emergence of on bean sprouts.Urea is the metabolic product of human normal, it is a kind of nitrogenous small-molecule substance, can urinate along with human body and discharge, a small amount of urea can't work the mischief to human body, when urea is assembled in a large number and can not be discharged, damage can be caused to human body, mainly make the cylinder electrolyte balance of human body destroyed, thus generation disease, i.e. uremia.These ammonium compounds in addition, under the effect of bacterium, can be transformed into nitrosamine, and nitrosamine are a kind of carcinogens.Strengthen the monitoring work to bud dish, seek the market demand that quick screening method is fast and accurately the screening of a large amount of sample.
At present, urea context of detection mainly contains several method:
1) Diacetylmonoxime development process
Diacetyl monoxime method principle is: strong acid heating condition under, urea and diacetyl react generate redness former diazine, shade and urea content proportional.This method reagent used is easy to get, and cost is low, but needs heating when detecting operation, and the range of linearity is low, is not generally suitable for field quick detection.The more important thing is that this kind of method is in bean sprouts urea measures, in bean sprouts, composition interference is comparatively large, easily causes false positive.
2) o-phthalaldehyde method
O-phthalaldehyde method measures urea principle: in acid condition, and the bond of urea and o-phthalaldehyde(OPA), under antipyrine existent condition, generates yellow compound, and the compound of this yellow and metavanadic acid root knot are closed in blue.This method does not need to boil, and simple to operate, disturbing factor is few, but the method reagent is not easily preserved for a long time, is unsuitable for make and examines product soon.
3) paradime thylaminobenzaldehyde method
Paradime thylaminobenzaldehyde method principle is: in an acidic solution, and urea and paradime thylaminobenzaldehyde generate the diaminobenzene formaldehyde urea of lemon yellow.The method stability, linearly good, interference less, easy to operate, but its sensitivity is not high, and the urea being generally used for the aspect such as dairy products, feed detects.
4) nitrite-Griess reagent method
Urea and nitrite react generation nitrogen under strongly acidic conditions, and Griess reagent and nitrite react and generate red compound.Reactant liquor redness containing urea can be partially light, and the color according to solution colour and titer judges whether to there is urea.It is qualitative that the method can be used for urea, and the sensitivity of reaction is 0.2%, does not reach the mensuration requirement of urea in bean sprouts.
Consider the defect of the aspects such as above method reagent stability, material interference, sensitivity, convenience, find a kind of easy and simple to handle, highly sensitive, result is accurate, cost is low, practical bean sprouts carbamide detection method is fast a large amount of sample screening in food security aspect market demand.Urase is a kind of urea decomposition catalyzer efficiently, urea generates NH4+ through urase enzymolysis, and according to ammonium radical ion hydrolysis PH change, the acid base indicator that causes is as variable colors such as cresol red, adjacent phenyl methylcarbamate complexing agent, methyl thymol blues or to survey NH4+ to urea indirect quantification by methods such as salicylic acid-indophenol blue, phenol-indophenol blues be one of study hotspot.
Acid base indicator Indirect Determination and liquid PH to be measured, damping fluid surge capability etc. have very large relation, and its detection is limited to more than 20mg/L urea.Salicylic acid-indophenol blue, phenol-ammonia-nitrogen principle is that sodium nitroprusside catalyzing N H4+ and phenol and hypochlorite react and generate blue compound, according to shade to urea indirect quantification, this kind of method is highly sensitive, but the reaction time is longer, is unsuitable for do and sieves product soon.And Na's reagent and NH4+ are swift in response, get final product colorimetric after shaking up, urase is combined with nessler reagent indirectly to be surveyed urea content and substantially increases detection sensitivity and fast screen capacity.
Summary of the invention
The object of the invention is the defect overcoming the aspects such as above method reagent stability, material interference, sensitivity, convenience, provide a kind of easy and simple to handle, highly sensitive, result is accurate, cost is low, practicality urea and total ammonia nitrogen detection method thereof in bean sprouts fast.
Technical scheme of the present invention is:
A detection method for urea and total ammonia nitrogen in bean sprouts, it, by detecting the ammonia nitrogen produced through urase decomposing urea, carries out sxemiquantitative according to ammonia-nitrogen content to urea, and its method detecting urea and total ammonia nitrogen comprises the following steps:
1) detecting thing is bean sprouts, is shredded bean sprouts, adds water and mixes, and room temperature is placed, and centre is shaken frequently, and obtaining supernatant is liquid to be measured;
2) get two parts respectively and detect liquid, portion adds urase, and another part adds isopyknic water, shakes up, room temperature catalysis;
3) to step 2) described in two parts of reactant liquors in add nessler reagent respectively, shake up;
4) step 3) shake up, observing response liquid color, contrasts with color scale card, to urea content sxemiquantitative.
Urase is beans urase; Described beans urase extracts from soya bean, green soya bean, black soya bean.
The preparation method of described beans urase is: in beans, add water, soaked overnight, and add ethanol water making beating, centrifugal, supernatant is beans urase liquid, and supernatant vacuum freeze drying is obtained beans urase.
Step 1) ratio of bean sprouts and water is 1:1-1:10.
Step 1) room temperature standing time is 2-15min.
Step 2) sample and urase amount ratio be 0.5mL:20-200uL.
Step 2) urease activity is 30 ~ 180UmL
-1.
Step 2) urase catalysis time is 2-10min.
Step 3) consumption of developer is 20-200uL.
Step 4) shake up after, contrasts with color scale card in 0-10min.
Step 4) contrast with color scale card, in liquid to be measured urea content for add urase group and do not add urase group with the difference of colour atla respective value, NH in liquid to be measured
4the content of-N is do not add urase group and color scale card contrasts the numerical value * 0.5 obtained.
Embodiment
Process when the present embodiment is mainly enumerated using soya bean urase as detection reagent.
Bean sprouts urea and total ammonia nitrogen quick detection reagent comprise urase and developer.
Urase is soya bean urase.
The preparation method of urase: soybean soaking is spent the night, adds ethanol water in soya bean, and pulverize, centrifugal, supernatant is soya bean urase liquid, and supernatant freeze drying is obtained soya bean urase.
Developer is made up of following mass percent: NaOH 16%, mercuric iodixde 10%, potassium iodide 7%, and surplus is water.
The method for quick of bean sprouts urea and total ammonia nitrogen, its step is as follows:
1) detecting thing is bean sprouts, is shredded bean sprouts, adds water and mixes, and room temperature places 2-15min, and centre is shaken frequently, and obtaining supernatant is liquid to be measured;
2) get two parts respectively and detect liquid, portion adds urase, and another part adds isopyknic water, shakes up, room temperature catalysis 2-10min;
3) to step 2) add developer in reactant liquor, shake up;
4) step 3) shake up, in 0-10min, observing response liquid color, contrasts with color scale card, to urea content sxemiquantitative.
Step 1) ratio of bean sprouts and water is 1:1-1:10.
Step 2) sample and urase amount ratio be 0.5mL:20-200uL.
Step 2) urease activity is 30 ~ 180UmL-1.
Step 3) consumption of developer is 20-200uL.
Step 4) contrast with color scale card, in liquid to be measured urea content for add urase group and do not add urase group with the difference of colour atla respective value, do not add urase group and color scale card contrasts the content that the numerical value * 0.5 obtained is NH4-N in liquid to be measured.
Above to theme disposal route and the device of a kind of resource provided by the present invention, be described in detail, apply specific case herein to set forth principle of the present invention and embodiment, the explanation of above embodiment just understands method of the present invention and core concept thereof for helping; Meanwhile, for one of ordinary skill in the art, according to thought of the present invention, all will change in specific embodiments and applications, in sum, this description should not be construed as limitation of the present invention.
What finally illustrate is, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (11)
1. the detection method of urea and total ammonia nitrogen in bean sprouts, is characterized in that: it, by detecting the ammonia nitrogen produced through urase decomposing urea, carries out sxemiquantitative according to ammonia-nitrogen content to urea, and its method detecting urea and total ammonia nitrogen comprises the following steps:
1) detecting thing is bean sprouts, is shredded bean sprouts, adds water and mixes, and room temperature is placed, and centre is shaken frequently, and obtaining supernatant is liquid to be measured;
2) get two parts respectively and detect liquid, portion adds urase, and another part adds isopyknic water, shakes up, room temperature catalysis;
3) to step 2) described in two parts of reactant liquors in add nessler reagent respectively, shake up;
4) step 3) shake up, observing response liquid color, contrasts with color scale card, to urea content sxemiquantitative.
2. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: urase is beans urase; Described beans urase extracts from soya bean, green soya bean, black soya bean.
3. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 2, it is characterized in that: the preparation method of described beans urase is: in beans, add water, soaked overnight, add ethanol water making beating, centrifugal, supernatant is beans urase liquid, and supernatant vacuum freeze drying is obtained beans urase.
4. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 1) ratio of bean sprouts and water is 1:1-1:10.
5. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 1) room temperature standing time is 2-15min.
6. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 2) sample and urase amount ratio be 0.5mL:20-200uL.
7. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 2) urease activity is 30 ~ 180UmL
-1.
8. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 2) urase catalysis time is 2-10min.
9. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 3) consumption of developer is 20-200uL.
10. the detection method of urea and total ammonia nitrogen in a kind of bean sprouts according to claim 1, is characterized in that: step 4) shake up after, contrasts with color scale card in 0-10min.
The detection method of urea and total ammonia nitrogen in 11. a kind of bean sprouts according to claim 1, it is characterized in that: step 4) contrast with color scale card, in liquid to be measured urea content for add urase group with do not add urase group with the difference of colour atla respective value, NH in liquid to be measured
4the content of-N is do not add urase group and color scale card contrasts the numerical value * 0.5 obtained.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582321.1A CN105203532A (en) | 2015-09-15 | 2015-09-15 | Detection method for urea and total ammonia nitrogen in bean sprouts |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582321.1A CN105203532A (en) | 2015-09-15 | 2015-09-15 | Detection method for urea and total ammonia nitrogen in bean sprouts |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105203532A true CN105203532A (en) | 2015-12-30 |
Family
ID=54951326
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510582321.1A Pending CN105203532A (en) | 2015-09-15 | 2015-09-15 | Detection method for urea and total ammonia nitrogen in bean sprouts |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105203532A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018360A (en) * | 2016-05-13 | 2016-10-12 | 深圳职业技术学院 | Method for detecting urea based on metal organic framework material fluorescent sensor |
| CN106501248A (en) * | 2016-11-06 | 2017-03-15 | 东北师范大学 | A kind of method of urea in high-throughput enzyme sensor and detection human urine |
| CN112858276A (en) * | 2021-01-25 | 2021-05-28 | 郑州欧柯奇仪器制造有限公司 | Soybean milk maturity rapid test card and detection method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4176008A (en) * | 1978-02-24 | 1979-11-27 | Eastman Kodak Company | Method, composition and element for the detection of nitrogen-containing compounds |
| CN101598727A (en) * | 2009-07-09 | 2009-12-09 | 上海科华生物工程股份有限公司 | The drying chemical reagent paper of quantitatively determining human blood urea content |
| CN102776266A (en) * | 2012-07-26 | 2012-11-14 | 广州绿洲生化科技股份有限公司 | Rapid testing card for heavy metal mercury and testing method thereof |
| CN103411965A (en) * | 2013-08-02 | 2013-11-27 | 福建农林大学 | Method for conveniently and quickly determining nitrogen content of plant material |
| CN104480092A (en) * | 2014-12-24 | 2015-04-01 | 陕西嘉禾植物化工有限责任公司 | Method for extracting urease from jack beans |
| CN104535711A (en) * | 2014-12-25 | 2015-04-22 | 广东环凯微生物科技有限公司 | Rapid detection kit and method of urea |
-
2015
- 2015-09-15 CN CN201510582321.1A patent/CN105203532A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4176008A (en) * | 1978-02-24 | 1979-11-27 | Eastman Kodak Company | Method, composition and element for the detection of nitrogen-containing compounds |
| CN101598727A (en) * | 2009-07-09 | 2009-12-09 | 上海科华生物工程股份有限公司 | The drying chemical reagent paper of quantitatively determining human blood urea content |
| CN102776266A (en) * | 2012-07-26 | 2012-11-14 | 广州绿洲生化科技股份有限公司 | Rapid testing card for heavy metal mercury and testing method thereof |
| CN103411965A (en) * | 2013-08-02 | 2013-11-27 | 福建农林大学 | Method for conveniently and quickly determining nitrogen content of plant material |
| CN104480092A (en) * | 2014-12-24 | 2015-04-01 | 陕西嘉禾植物化工有限责任公司 | Method for extracting urease from jack beans |
| CN104535711A (en) * | 2014-12-25 | 2015-04-22 | 广东环凯微生物科技有限公司 | Rapid detection kit and method of urea |
Non-Patent Citations (4)
| Title |
|---|
| 周东凯 等: "大豆脲酶的提取及其影响因素研究", 《大豆科学》 * |
| 林丽云 等: "黄豆脲酶的提取及其活性测定", 《食品研究与开发》 * |
| 王武生 等: "酶解法检测牛奶中的尿素", 《食品科学》 * |
| 齐国艳 等: "绿豆芽中尿素的定性、定量测定方法探讨", 《中国卫生检验杂志》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018360A (en) * | 2016-05-13 | 2016-10-12 | 深圳职业技术学院 | Method for detecting urea based on metal organic framework material fluorescent sensor |
| CN106018360B (en) * | 2016-05-13 | 2018-07-17 | 深圳职业技术学院 | A method of urea is detected based on metal-organic framework materials fluorescent optical sensor |
| CN106501248A (en) * | 2016-11-06 | 2017-03-15 | 东北师范大学 | A kind of method of urea in high-throughput enzyme sensor and detection human urine |
| CN106501248B (en) * | 2016-11-06 | 2018-12-04 | 东北师范大学 | A kind of method of urea in high-throughput enzyme sensor and detection human urine |
| CN112858276A (en) * | 2021-01-25 | 2021-05-28 | 郑州欧柯奇仪器制造有限公司 | Soybean milk maturity rapid test card and detection method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Colorimetric array indicator for NH3 and CO2 detection | |
| CN105203532A (en) | Detection method for urea and total ammonia nitrogen in bean sprouts | |
| CN101900684A (en) | Test paper for quickly detecting urea in liquid milk and preparation method thereof | |
| CN104502515B (en) | A kind of LC-MS/MS assay method of four chlorantraniliprole residual quantities | |
| CN101398386A (en) | Preparation of carbamide detection test paper for dairy food and carbamide detection method | |
| CN103698324A (en) | Air formaldehyde self testing kit | |
| CN103776824A (en) | Quick detection method and detection kit for hydrazoate in feed | |
| Cancellier et al. | Rapid and low-cost method for evaluation of nutrient release from controlled-release fertilizers using electrical conductivity | |
| Thongprajukaew et al. | Smart phone: a popular device supports amylase activity assay in fisheries research | |
| CN104076034A (en) | RGB quantitative quick detecting test paper for formaldehyde residue in food | |
| US6764856B2 (en) | Semen detection system and method | |
| CN205719999U (en) | Formaldehyde in food Test paper | |
| Zhou et al. | Modeling and prediction for the acute toxicity of pesticide mixtures to the freshwater luminescent bacterium Vibrio qinghaiensis sp.-Q67 | |
| CN105294510B (en) | A kind of stable isotope13C or15N marks the synthetic method of biruea | |
| CN107764815A (en) | The method of quick detection Nitrate Content in Food | |
| CN104297177A (en) | Detection kit for protein in dairy products and detection method | |
| CN106226487A (en) | The method utilizing the metallothionein detection degree of water pollution of fish | |
| CN107703296B (en) | Ultra-sensitive electrochemical immunosensor for multi-residue detection of sulfonamides | |
| KR101226103B1 (en) | Method of detecting leachate from livestock disposal landfill area | |
| Ye et al. | Use of elemental fingerprint analysis to identify localities of collection for the large icefish protosalanx chinensis in Taihu Lake, China | |
| CN104034721A (en) | Detection method of ammoniacal nitrogen in fermenting liquid | |
| US20240118296A1 (en) | Quantitative pooled-sample testing method and apparatus for chemical test items of consumer product | |
| CN106323960A (en) | Quick detection method and detection kit of ammonium nitrogen in fertilizer | |
| WO2021112694A2 (en) | System and method for detecting analyte in food sample | |
| Jin et al. | A stable isotope and chemometric framework to distinguish fresh milk from reconstituted milk powder and detect potential extraneous nitrogen additives |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151230 |