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High sensitivity detection method for cowhide cultural relic

A technology with high sensitivity and detection method, applied in the field of high-sensitivity detection of cowhide cultural relics, can solve problems such as difficulty in detection of cultural relics with extremely low protein content, and achieve the effects of high binding rate, high utilization efficiency and good load performance.

Active Publication Date: 2019-03-29
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current advanced technology for protein detection is based on antibody-antigen Western Blotting or ELISA, but these methods are still difficult to detect some artifacts with extremely low protein content

Method used

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  • High sensitivity detection method for cowhide cultural relic
  • High sensitivity detection method for cowhide cultural relic

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A) Take 15 g of healthy adult cattle hide, cut it into small pieces of 2 cm × 2 cm, put it into a conical flask, soak and degrease it with 5% sodium carbonate aqueous solution in a 45°C incubator for 4 h. Take out the degreased cowhide, mix each 1 g of cowhide with 14 mL of 4% sodium chloride aqueous solution, stir slowly at room temperature for 12 h to remove the salt-soluble non-collagen components, and then wash and drain with distilled water.

[0041] B) Put the drained cowhide in a beaker, add pepsin, the amount of protease added is 2000 U / g cowhide, add 28 mL of 0.5 mol / L acetic acid to 1 g cowhide, and carry out enzymatic hydrolysis in a 37 ℃ incubator Reaction for 7 h. After enzymatic hydrolysis, filter, collect the filtrate, add sodium chloride for salting out, the concentration of sodium chloride is 3 mol / L, after salting out, centrifuge at 12000 r / min for 10 min, and dissolve the precipitate with 0.5 mol / L acetic acid Dialysis bags with a cutoff of 10,000 we...

Embodiment 2

[0055] A) Take 15 g of healthy adult cattle hide, cut it into small pieces of 2 cm × 2 cm, put it into a conical flask, soak and degrease it with 6% sodium carbonate aqueous solution in a 50°C thermostat for 4 h. Take out the degreased cowhide, mix each 1 g of cowhide with 15 mL of 4-6% sodium chloride aqueous solution, stir slowly at room temperature for 12 hours to remove salt-soluble non-collagen components, and then wash and drain with distilled water.

[0056] B) Put the drained cowhide in a beaker, add pepsin, the amount of protease added is 3000 U / g cowhide, add 30 mL of 0.5 mol / L acetic acid to 1 g cowhide, and carry out enzymatic hydrolysis in a 37 ℃ incubator React for 8 h. After enzymatic hydrolysis, filter, collect the filtrate, add sodium chloride for salting out, the concentration of sodium chloride is 3-5 mol / L, centrifuge at 12000 r / min for 10 min after salting out, and dissolve the precipitate in 0.5 mol / L acetic acid Dialyze for 3 days in a dialysis bag with...

Embodiment 3

[0068] A) Take 15 g of healthy adult cattle hide, cut it into small pieces of 2 cm×2 cm, put it into a conical flask, soak and degrease it with 7% sodium carbonate aqueous solution in a 55°C incubator for 4 h. Take out the degreased cowhide, mix each 1 g of cowhide with 16 mL of 4-6% sodium chloride aqueous solution, stir slowly at room temperature for 12 h to remove the salt-soluble non-collagen components, and then wash and drain with distilled water.

[0069] B) Put the drained cowhide in a beaker, add pepsin, the amount of protease added is 4000 U / g cowhide, add 32 mL of 0.5 mol / L acetic acid to 1 g cowhide, and carry out enzymatic hydrolysis in a 37 ℃ incubator React for 9 h. After enzymatic hydrolysis, filter, collect the filtrate, add sodium chloride for salting out, the concentration of sodium chloride is 3-5 mol / L, centrifuge at 12000 r / min for 10 min after salting out, and dissolve the precipitate in 0.5 mol / L acetic acid Dialyze for 3 days in a dialysis bag with a ...

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Abstract

The invention relates to the cultural relic detection field, and discloses a high sensitivity detection method for a cowhide cultural relic. The method comprises the steps that pure bovine collagen powder is prepared, and bovine collagen in a cultural relic sample is extracted; the surface of a cleaned glassy carbon electrode is sequentially modified with gold nanoparticles, a polyacrylic acid-dopamine composite with a hollow structure, polymethyl methacrylate-maleic anhydride / 1-octadecene alternating copolymer spherical nano beads and a rabbit anti-bovine collagen polyclonal antibody under the effect of an NHS / EDC solution by utilizing a layer-by-layer assembly method; and a bovine collagen solution and a cultural relic sample solution are detected by means of the modified electrode. Themethod is high in sensitivity, low in detection lower limit, good in repeatability and capable of achieving supersensitive detection on the cowhide cultural relic.

Description

technical field [0001] The invention relates to the field of cultural relics detection, in particular to a high-sensitivity detection method for cowhide cultural relics. Background technique [0002] Humans have a very long history of using leather products. As early as in ancient times, our ancestors began to use leather made of various animal skins. In the thousands of years of historical precipitation, there are a large number of leather cultural relics left in the world. Leather is a natural polymer fiber, the main component of which is collagen protein. After a long period of preservation, the surface morphology of leather cultural relics is severely damaged, and it is very difficult to identify its species. Therefore, how to use advanced methods of natural science to establish a micro-mark detection technology system for leather cultural relics, and to extract ancient leather information from impressions, residues, and soil is crucial to the study of the origin and ...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/30
CPCG01N27/308G01N27/3277G01N27/3278
Inventor 李津王秉邓博之朱聘宇胡智文
Owner ZHEJIANG SCI-TECH UNIV
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