Method and reagent for determining hydroxyproline in feed

The technology of a hydroxyproline and a detection method, which is applied in the detection field of hydroxyproline, can solve the problems of long processing time and low detection specificity, and achieves the effects of simplifying the mobile phase formula, reducing the analysis time and reducing the detection cost.

Inactive Publication Date: 2018-05-08
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the problems of long sample pretreatment time and low detection specificity in the hydroxyproline determination method existing in the prior art, the purpose of the

Method used

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  • Method and reagent for determining hydroxyproline in feed
  • Method and reagent for determining hydroxyproline in feed
  • Method and reagent for determining hydroxyproline in feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1, the rapid detection method of hydroxyproline in feed

[0070] A. Sample pretreatment

[0071] Crush the feed sample to be tested, accurately weigh 100 mg of the sample, put it into a microwave hydrolysis tank, add 5 mL of 6mol / L hydrochloric acid solution, feed nitrogen, cover the tank cover, and follow the microwave hydrolysis instrument (Multiwave Pro microwave digestion instrument, Anton Paar Company, Austria) ) Condition setting and hydrolysis: control the microwave power to 600W, control the temperature to rise from room temperature to 150°C within 5 minutes, and then keep the temperature for 30 minutes; , add 3mL of 0.02mol / L hydrochloric acid solution, vortex to mix evenly, filter through a 0.45μm microporous membrane, and wait for detection by an amino acid analyzer;

[0072] B. Amino acid analyzer preparation

[0073] Amino acid analyzer chromatographic column: Hitachi L-8900 automatic amino acid analyzer, separation column 4.6mm×40mm; stationar...

Embodiment 2

[0094] The detection of hydroxyproline in embodiment 2, fish meal

[0095] Accurately weigh 100 mg of the fish meal sample to be tested and put it into a microwave hydrolysis tank, add 5 mL of 6mol / L hydrochloric acid solution, blow in nitrogen gas, cover the tank cover, set up and perform hydrolysis according to the conditions of the microwave hydrolysis instrument: the microwave power is controlled at 600W, Control the temperature to rise from room temperature to 150°C within 5 minutes, and then keep the temperature for 30 minutes; after the constant temperature is over, pipette 200 μL of hydrolyzed solution to the concentrated hydrolysis tube, concentrate and evaporate to dryness with a concentrator, add 3 mL of 0.02 mol / L hydrochloric acid solution, and vortex to mix Evenly, through 0.45 μm microporous membrane filtration, measure 3 groups of parallel experiments with the method for embodiment 1, the result is averaged, and measurement result is as follows figure 2 .

Embodiment 3

[0096] Embodiment 3, the detection of hydroxyproline in meat and bone meal

[0097] Accurately weigh 100mg of the meat and bone meal sample to be tested and put it into a microwave hydrolysis tank, add 5mL of 6mol / L hydrochloric acid solution, blow in nitrogen gas, cover the tank lid, set up and perform hydrolysis according to the conditions of the microwave hydrolysis instrument: the control microwave power is 600W , control the temperature to rise from room temperature to 150 °C within 5 minutes, and then keep the temperature for 30 minutes; after the constant temperature is over, pipette 200 μL of hydrolyzate to the concentrated hydrolysis tube, concentrate and evaporate to dryness with a concentrator, add 3 mL of 0.02 mol / L hydrochloric acid solution, and vortex Mix homogeneously, filter through 0.45 μm microporous membrane, measure 3 groups of parallel experiments with the method of embodiment 1, the result is averaged, and measurement result is as follows: image 3 .

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Abstract

The invention relates to a method and reagent for determining hydroxyproline in a feed. The method comprises the following steps: preprocessing a to-be-measured sample, and hydrolyzing a protein in the to-be-measured sample; determining the content of hydroxyproline in a hydrolysate of the protein by using an ion exchange chromatography method, wherein a first eluant used for separation with a cation exchange column is a solution with the solvent of water and solutes and the concentration respectively comprising 0.02-0.03 mol/L of sodium citrate, 0.075-0.125 mol/L of sodium chloride, 0.075-0.125 mol/L of citric acid, 12-16% (volume fraction) of ethanol. With improvement of preparation of mobile phase reagents and the instrumental analysis method, the method is suitable for rapid detectionof hydroxyproline. The method can separate hydroxyproline from other amino acids completely and have rapid and accurate characteristics. The method can accurately analyze hydroxyproline in the feed, can be used for large-scale sample detection and is suitable for standardization.

Description

technical field [0001] The invention relates to the technical field of detection of hydroxyproline, in particular to a detection method and a reagent for hydroxyproline in feed. Background technique [0002] Hydroxyproline, formula C 5 h 9 NO 3 , Molecular weight 131.13, also known as trans-4-hydroxy-L-proline, is a unique amino acid of collagen, the content in collagen is about 13.4%, the content in elastin is very small, and it does not exist in other proteins . [0003] Animal collagen hydrolyzed protein is a hydrolyzed protein made by rough processing of animal fur and its product waste. It can be used as animal feed and is a cheap protein raw material. However, in order to increase the protein content of dairy products and reduce production costs, some illegal manufacturers have mixed cheap hydrolyzed animal protein into milk powder to pretend to replace milk protein. The added animal collagen hydrolyzed protein will not only affect the taste and flavor of dairy pro...

Claims

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Application Information

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IPC IPC(8): G01N30/96
CPCG01N30/96
Inventor 贾铮樊霞李兰赵根龙
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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