Quantitative determination method in extraction process of fish collagen and application of quantitative determination method

A quantitative detection method and technology for fish collagen, which are applied in the preparation of test samples and the measurement of color/spectral properties, etc., can solve the problems of tediousness and long time for quantitative collagen detection, and achieve the effect of good stability.

Inactive Publication Date: 2014-05-07
SOUTH CHINA UNIV OF TECH
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004] In order to overcome the shortcomings and deficiencies such as long detection time and tediousness in the quantitative collagen of the above-mentioned prior art, the primary purpose of the present invention is to provide a rapid quantitative detection method for fish collagen content in the extraction process

Method used

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  • Quantitative determination method in extraction process of fish collagen and application of quantitative determination method
  • Quantitative determination method in extraction process of fish collagen and application of quantitative determination method
  • Quantitative determination method in extraction process of fish collagen and application of quantitative determination method

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1: Determination of collagen content in fish scales extracted by pepsin

[0029] (1) Drawing of the standard curve: the collagen standard (rat skin type I collagen) was dissolved in 0.5M acetic acid and diluted to a standard solution containing 1-300ug collagen per 100ul. Take 11 copies of collagen standard solution, each 0.1mL, the collagen content is 1ug, 3ug, 5ug, 10ug, 20ug, 30ug, 50ug, 100ug, 150ug, 200ug and 300ug. Add 1ml 69mg / L Sirius Scarlet solution to each portion, mix well, let stand at room temperature for 30min, then centrifuge at 11000r / min for 30min, collect the supernatant, redissolve the precipitate with the same concentration of acetic acid, and centrifuge under the same conditions to obtain the above Serum. Combine the centrifuged supernatants twice, and measure the absorbance of the supernatants at 540 nm. Finally, the standard curve of the supernatant was drawn with the absorbance as the ordinate and the collagen concentration as the a...

Embodiment 2

[0033] Embodiment 2: Determination of collagen content in fish scales after acetic acid extraction

[0034] Standard curve is drawn with embodiment 1.

[0035] Prepare the collagen extracted by acetic acid at 4°C into a saturated 0.5M acetic acid solution, add 69 mg / L Sirius red solution according to the dosage of 1:10, stir well, let stand at room temperature for 15 minutes for staining, centrifuge, read Take the absorbance at 540nm, check the standard curve, and obtain the corresponding collagen concentration, the result is as follows image 3 .

Embodiment 3

[0036] Embodiment 3: Fish scale is measured in the amount of loss of collagen in EDTA decalcification process

[0037] Get 100mL of 0.05mol / L, 0.10mol / L, 0.15mol / L, 0.20mol / L and 0.25mol / L of EDTA decalcification solution and mix with 2g fish scale collagen respectively, react 120min, according to the measuring procedure of embodiment 1, use The content of collagen after decalcification was measured by Sirius scarlet staining, and the amount of collagen loss during decalcification was converted. The results are shown in Table 2.

[0038] The amount of loss of collagen in the process of table 2 decalcification

[0039]

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Abstract

The invention belongs to the technical field of aquatic product processing and discloses a rapid quantitative determination method in an extraction process of fish collagen and an application of the quantitative determination method. The method comprises the following steps: preparing a collagen liquid by using an acetic acid liquid; after dyeing by using sirius red, centrifuging and determining the absorbance of liquid supernatant; comparing a standard curve; and quantifying the content of the collagen. According to the quantitative determination method, the specific binding of the sirius red and the collagen is utilized; after the centrifuging, the absorbance of the liquid supernatant can be determined under the condition of a special wavelength of 540nm; and the content of the collagen is determined. According to the quantitative determination method, the centrifuged liquid supernatant is determined so that a solvent is saved and the influences on specific matters formed by the collagen and the sirius red, caused by the liquid and other reagents, are avoided; the recovery rate of a sample is 95.7-104.7% and the requirements of analytical researches are met; the variable coefficient of a parallel sample is 0.35% and the stability is very good; and the quantitative determination method is applied to quantifying the collagen in an operation process and the present determination can be realized.

Description

technical field [0001] The invention belongs to the technical field of aquatic product processing, in particular to a rapid quantitative detection method in the process of extracting fish collagen and its application. Background technique [0002] Fish scale (skin) is rich in collagen, extracting collagen from fish scale (skin) can make full use of fish scale waste and turn waste into treasure. Therefore, it is of great significance to measure the change of collagen content during the extraction process to study the functionality and product performance of fish scales (skins). [0003] There are many methods for quantitative analysis of collagen or collagen. For samples known to be collagen or collagen, Kjeldahl method is the most reliable, followed by biuret method, UV spectrophotometry, Folin-phenol method and Coomassie Brilliant Blue G-250 staining, etc. However, for samples that are not known to be collagen or collagen, the above method is not suitable. The convention...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N1/28
Inventor 朱志伟梁健华曾庆孝李汴生张颖洁
Owner SOUTH CHINA UNIV OF TECH
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