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Method for Determination of Purine by Ultraviolet Spectrophotometer

A spectrophotometer and ultraviolet technology, which is applied in the field of animal science, can solve problems such as complex components, and achieve the effects of high repeatability, accurate values, and easy operation

Inactive Publication Date: 2011-12-14
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microorganisms in the freeze-dried sample of rumen microorganisms exist in the form of bacteria, and at the same time contain other substances, and the composition is relatively complex

Method used

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  • Method for Determination of Purine by Ultraviolet Spectrophotometer
  • Method for Determination of Purine by Ultraviolet Spectrophotometer
  • Method for Determination of Purine by Ultraviolet Spectrophotometer

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1: a kind of ultraviolet spectrophotometer of the present invention measures the method for purine, and the steps are as follows:

[0019] Step 1: Accurately weigh 0.5 g of fully dried (dried with a freeze dryer) chyme sample, or a sample of the isolated chyme microbial part, and place it in a 25ml test tube with stoppers and scales. Note: the sample must be dry , otherwise it will lead to incomplete nucleic acid hydrolysis;

[0020] Step 2: add 2.5ml perchloric acid (HClO 4 , 1M), tightly cap the bottle mouth, and then place it in a constant temperature water bath at 90-95°C for 1 hour and incubate for 1 hour. At this time, the sample is carbonized and black indurations are formed. Break the indurations to facilitate the reaction to complete;

[0021] Step 3: Add 17.5ml of acetic acid buffer (NH 4 h 2 PO 4 , 0.0285M), fully shake, tightly cap the bottle, put it in a constant temperature water bath at 90-95°C for 10-15 minutes, and then filter through Wha...

Embodiment 2

[0030] Embodiment 2: the method that the ultraviolet spectrophotometer of the present invention measures purine is the purine recovery rate ultraviolet spectrophotometer method, and the purine recovery rate ultraviolet spectrophotometer method and liquid chromatography comparative study are as follows:

[0031] 1 Preparation of samples

[0032] 1.1 Preparation of rumen microbial freeze-dried samples

[0033] Take 1 L of rumen content 2 hours after morning feeding. Filter through two layers of cotton cloth and store at 4°C filled with CO 2 1L jar with a stopper for 30 minutes. Use a pipette to suck up a homogeneous liquid in the middle that does not contain the heavier particles in the lower layer and the lighter particles in the upper layer. Centrifuge at 20,000×g for 20 minutes at 4°C. The spheres were washed with distilled water, centrifuged, and finally freeze-dried.

[0034] 1.2 Preparation of neutral detergent fiber

[0035] Medium-sized detergent fibers were prepar...

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Abstract

The invention provides a method for measuring purine with an ultraviolet spectrophotometer. The steps include: accurately weighing 0.5 g of fully dried (dried by freeze dryer) chyme sample, and placing it in a 25 ml test tube with stopper and scale; adding 2.5 ml of perchloric acid, and placing it in a constant temperature water bath at 90-95 °C Incubate for 1 hour; then add 17.5ml of acetic acid buffer, place it again in a constant temperature water bath at 90-95°C and incubate for 10-15 minutes, then filter; transfer 0.5ml of the filtrate to a centrifuge tube with a volume of 15ml, and centrifuge at 4000rpm for 15 minutes ; be placed in a constant temperature water bath of 90-95 ℃ and cultivated for 30 minutes; use a UV spectrophotometer to compare color at 260nm, and the measured purine concentration is represented by RNA equivalent; the invention is easy to operate, high in repeatability, and accurate in value Spectrophotometer was used to measure the content of purine in digesta and the content of microbial protein in duodenal digesta, which provided a methodological basis for determining the content of metabolic protein in dairy cows.

Description

(1) Technical field [0001] The invention relates to animal science, in particular to a method for measuring purine with an ultraviolet spectrophotometer. (2) Background technology [0002] The dietary duodenal metabolizable protein (Metablizable protein, MP) of dairy cows includes dietary rumen undegradable protein (RUP), rumen microbial protein (Microbial crude protein, MCP) and endogenous protein (Endogenous crude protein, ECP) three parts. Rumen microbial protein accounts for 60-80% of metabolic protein, so quantification of rumen microbial protein synthesis is very important. The purine nitrogen method proposed by Zinn and Owen (1986) has been adopted by some scholars because of its simplicity, quickness, low test cost and easy operation, but it has also been questioned by some scholars because of the relatively low recovery rate of purines (Perez et al, 1997 ; Makker et al, 1999; Gonzalaz et al, 2003, 2004). This method is based on the hydrolysis of rumen microorgani...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33
Inventor 单安山乔国华
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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