CN105203478A - Fast detection method of leather-hydrolyzed protein L-hydroxyproline - Google Patents
Fast detection method of leather-hydrolyzed protein L-hydroxyproline Download PDFInfo
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- CN105203478A CN105203478A CN201510585318.5A CN201510585318A CN105203478A CN 105203478 A CN105203478 A CN 105203478A CN 201510585318 A CN201510585318 A CN 201510585318A CN 105203478 A CN105203478 A CN 105203478A
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- hydroxyproline
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Abstract
The invention discloses a fast detection method of leather-hydrolyzed protein L-hydroxyproline, in particular to a fast detection method of leather-hydrolyzed protein L-hydroxyproline illegally added into milk. The fast detection method of the leather-hydrolyzed protein L-hydroxyproline illegally added into the milk is convenient to use, fast, simple and convenient to operate and high in sensitivity and is characterized in that a reagent is added into the milk or a dairy product to be detected and is heated in water bath to conduct precipitation treatment on protein in the milk or the dairy product, and standing is performed for layering; an oxidizing agent and a color developing agent are added into supernatant collected in the step 1, reaction is performed for a period of time, and then a colorimetric method is adopted to determine the L-hydroxyproline so as to obtain related results.
Description
Technical field
The invention belongs to spectrophotometry field, relate to a kind of detection method, be related specifically to a kind of method for quick of leather milk protein hydrolysate L-hydroxyproline.
Background technology
So-called " leather protein hydrolysate ", utilizes leather-solid waste or the fur of animal, internal organs etc., a kind of albumen powder generated through technology for hydrolyzing exactly.The same with melamine, it is mixed the content that can improve protein in milk or milk powder.The Ministry of Agriculture to the detection whether containing " leather aminosal " in fresh milk goods be not this year inchoate.After " melamine milk " event, the Ministry of Agriculture started to carry out the monitoring of fresh milk quality safety in China in 2009, and the content of monitoring comprises melamine, leather aminosal " and alkaloids.On January 5th, 2011, the Ministry of Agriculture has issued " 2011 annual fresh milk quality of item safety monitoring plan ", wherein require monitor project and unchanged.Its detection method comprises colourimetry, electrophoresis high performance liquid chromatography etc.At present conventional colourimetry (colorimetry) is by relatively or measure the method that the coloring matter solution colour degree of depth determines component concentration to be measured.Colorimetric determination reagent toxicity is high, operates relatively loaded down with trivial details.
Summary of the invention
For above-mentioned deficiency, the object of the present invention is to provide a kind of reagent avoiding using toxicity high, troublesome operation simplified, can detect the content of sample fast and effectively, this project detection method sampling amount is few, and detection time is not long, and technique is simple.
For solving the problems of the technologies described above, technical scheme provided by the invention is such:
A method for quick for leather milk protein hydrolysate L-hydroxyproline, comprises the steps: successively
1) pre-treatment reagent process is added to the product that need detect;
2) get step 1) in supernatant solution, add oxidation buffer solution and chromogenic reagent;
3) by step 2) in solution put into cuvette, adopt colourimetry L-hydroxyproline is wherein measured thus obtains correlated results.
Further, step 1) described in pre-treatment reagent be 219g/L zinc acetate+30mL/L glacial acetic acid and 106g/L potassium ferrocyanide.
Further, described oxidation buffer solution is PBS and toluene-sodium-sulfonchloramide, and toluene-sodium-sulfonchloramide concentration is 1.2 ~ 1.7g/100mL.
Further, described developer is paradime thylaminobenzaldehyde.
The invention has the beneficial effects as follows: provide a kind of reagent avoiding using toxicity high, troublesome operation simplified, can detect the content of sample fast and effectively, this project detection method sampling amount is few, and detection time is not long, and technique is simple.
Embodiment
Embodiment
The present embodiment introduces the compound method of solution and the reagent used in the present invention and detects the flow and method of operation.
1), pre-treatment reagent is prepared: accurately take 219g zinc acetate and 30mL glacial acetic acid, with distilled water constant volume to 1000mL, stirring and dissolving.Accurately take 106g potassium ferrocyanide, with distilled water constant volume to 1000mL, stirring and dissolving.
2), oxidation buffer liquid is prepared: take 6.95g ADSP, 8g sodium dihydrogen phosphate two water, 7.05g toluene-sodium-sulfonchloramide respectively, with distilled water constant volume to 500mL, stirring and dissolving.
3), developer is prepared: accurately measure 100mL concentrated hydrochloric acid and slowly add in 100mL water; Take 50g paradime thylaminobenzaldehyde, measure the 175mL concentrated hydrochloric acid solution after above-mentioned dilution, stirring and dissolving, and then add 325mL isopropyl alcohol, stir and make it mix.This reagent is oxidizable, needs airtight Cord blood.
4), standard solution preparation: the L-hydroxyproline adding distil water in the volumetric flask of 100mL getting 0.1g dissolves, and dripping the sulfuric acid solution mixing of a 3mol/L, with distilled water constant volume to 100mL, is the mark liquid of 1000ppm.(this mark liquid should be placed on 4 DEG C of stored refrigerated)
5), detection method: get 2-5g sample and add 30mL water, 60 DEG C of heating water bath 20min; Be cooled to room temperature; Add 1mL acetic acid zinc solution and 1mL potassium ferrocyanide solution respectively, be settled to 50mL with water, centrifugal, get supernatant (if solution is muddy, get after filtration filtrate to be measured) to be measured.Get the centrifuge tube of a 2mL, add 1mL sample liquid.Add 500 μ L oxidation buffer liquid, lid lid turns upside down and makes for three times it even, and add 500 μ L developers again after reaction 2min, lid lid shakes up.Heat 7 minutes in 70 DEG C of water-baths, after taking-up, be cooled to room temperature.Take distilled water as blank, 536nm measures absorbance.Absorbance is substituted into regression equation and obtains hydroxyproline content.
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 (4)
1. a method for quick for leather milk protein hydrolysate L-hydroxyproline, is characterized in that, comprise the steps: successively
1) pre-treatment reagent process is added to the product that need detect;
2) get step 1) in supernatant solution, add oxidation buffer solution and chromogenic reagent;
3) by step 2) in solution put into cuvette, adopt colourimetry L-hydroxyproline is wherein measured thus obtains correlated results.
2. the method for quick of a kind of leather milk protein hydrolysate L-hydroxyproline according to claim 1, is characterized in that, step 1) described in pre-treatment reagent be 219g/L zinc acetate, 30mL/L glacial acetic acid and 106g/L potassium ferrocyanide.
3. the method for quick of a kind of leather milk protein hydrolysate L-hydroxyproline according to claim 1, it is characterized in that, described oxidation buffer solution is PBS and toluene-sodium-sulfonchloramide, and toluene-sodium-sulfonchloramide concentration is 1.2 ~ 1.7g/100mL.
4. the method for quick of a kind of leather milk protein hydrolysate L-hydroxyproline according to claim 1, it is characterized in that, described developer is paradime thylaminobenzaldehyde.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510585318.5A CN105203478A (en) | 2015-09-15 | 2015-09-15 | Fast detection method of leather-hydrolyzed protein L-hydroxyproline |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510585318.5A CN105203478A (en) | 2015-09-15 | 2015-09-15 | Fast detection method of leather-hydrolyzed protein L-hydroxyproline |
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| CN105203478A true CN105203478A (en) | 2015-12-30 |
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| CN201510585318.5A Pending CN105203478A (en) | 2015-09-15 | 2015-09-15 | Fast detection method of leather-hydrolyzed protein L-hydroxyproline |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572069A (en) * | 2016-01-19 | 2016-05-11 | 江南大学 | Rapid detection method of cis-3-hydroxyL-proline in fermentation liquor |
| CN105651716A (en) * | 2016-02-06 | 2016-06-08 | 渭南市华隆畜牧有限公司 | Rapid ELISA (enzyme-linked immunosorbent assay) method for leather hydrolyzed protein L-hydroxyproline in raw milk |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02298865A (en) * | 1989-02-09 | 1990-12-11 | Seiichi Inayama | Rapid determination of hydroxyproline |
| CN101871893A (en) * | 2009-04-21 | 2010-10-27 | 陈延霞 | Colorimetric method for determining free L-hydroxyproline (L-HYP) in milk powder |
| CN103018185A (en) * | 2012-12-05 | 2013-04-03 | 青岛艾华隆生物科技有限公司 | Method for quickly measuring collagen content |
| CN103926356A (en) * | 2013-11-08 | 2014-07-16 | 中国计量科学研究院 | Quality control sample for leather hydrolyzed protein-containing milk powder and its preparation method |
| CN104359848A (en) * | 2014-12-10 | 2015-02-18 | 福建师范大学 | Method for measuring content of collagen |
-
2015
- 2015-09-15 CN CN201510585318.5A patent/CN105203478A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02298865A (en) * | 1989-02-09 | 1990-12-11 | Seiichi Inayama | Rapid determination of hydroxyproline |
| CN101871893A (en) * | 2009-04-21 | 2010-10-27 | 陈延霞 | Colorimetric method for determining free L-hydroxyproline (L-HYP) in milk powder |
| CN103018185A (en) * | 2012-12-05 | 2013-04-03 | 青岛艾华隆生物科技有限公司 | Method for quickly measuring collagen content |
| CN103926356A (en) * | 2013-11-08 | 2014-07-16 | 中国计量科学研究院 | Quality control sample for leather hydrolyzed protein-containing milk powder and its preparation method |
| CN104359848A (en) * | 2014-12-10 | 2015-02-18 | 福建师范大学 | Method for measuring content of collagen |
Non-Patent Citations (2)
| Title |
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| 刘学琴 等: "生鲜乳中添加皮革水解蛋白的测定方法研究", 《乳业科学与技术》 * |
| 赵玉富 等: "分光光度法测定乳及乳制品中L-羟脯氨酸的改进研究", 《中国乳品工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105572069A (en) * | 2016-01-19 | 2016-05-11 | 江南大学 | Rapid detection method of cis-3-hydroxyL-proline in fermentation liquor |
| CN105651716A (en) * | 2016-02-06 | 2016-06-08 | 渭南市华隆畜牧有限公司 | Rapid ELISA (enzyme-linked immunosorbent assay) method for leather hydrolyzed protein L-hydroxyproline in raw milk |
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