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Analytical detection method for effective identification of different species of soft-shelled soft-shelled turtle

A detection method, the technology of Chinese soft-shelled turtle, which is applied in the direction of analyzing materials, measuring devices, and material separation, can solve problems such as time-consuming, analysis and detection limitations, and demanding requirements, and achieve detection accuracy and effect improvement, high resolution and sensitivity. Effect

Active Publication Date: 2021-04-13
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these commonly used methods are time-consuming and expensive, and when the analyte is complex or there is DNA contamination, the analysis and detection will be greatly limited
Furthermore, specific probe sequences and associated DNA sequencing are required when the species under study is not present in the database, making genotyping methods for species identification technically demanding

Method used

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  • Analytical detection method for effective identification of different species of soft-shelled soft-shelled turtle
  • Analytical detection method for effective identification of different species of soft-shelled soft-shelled turtle
  • Analytical detection method for effective identification of different species of soft-shelled soft-shelled turtle

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, a kind of analytical detection method that effectively distinguishes different kinds of Chinese soft-shelled turtles, following steps successively:

[0046] 1) Take 0.2 g of the muscle tissue of soft-shelled turtle to be tested and grind it uniformly into a slurry, add 1.2 mL of dichloromethane-methanol mixed extraction solution, shake evenly at 30°C for 5 minutes, and obtain the extract (product A).

[0047] In the dichloromethane-methanol mixed extraction solution, dichloromethane:methanol=2:1 volume ratio (v / v);

[0048] 2) After extracting the extract (product A) at an extraction temperature of 50°C and 53KHz for 30 minutes with ultrasonic assistance, add 0.8mL of ultrapure water, and rapidly freeze and centrifuge at -20°C for 15 minutes.

[0049] 3), adding 2mL of dichloromethane to the first aqueous phase (upper layer) obtained in step 2) by refrigerated centrifugation for secondary extraction, respectively obtaining a secondary aqueous phase on the ...

Embodiment 2

[0075] Embodiment 2. The ultrasonic extraction temperature parameters in Step 2 of Embodiment 1 are set to 40°C, 50°C, and 60°C respectively; the rest are the same as in Embodiment 1. The comparison of the results of the obtained recovery rate is as follows figure 1 As described in A.

[0076] According to the figure 1 A, it can be known that the optimum extraction temperature is 50°C.

Embodiment 3

[0078] The sum of the amount of the dichloromethane-methanol mixed extraction solution in step 1) of Example 1 and the ultrapure water in step 2) is set to 0.5mL, 1mL, 2mL and 3mL respectively (dichloromethane-methanol mixed extraction solution : volume ratio of ultrapure water=1.5:1); the rest are equal to embodiment 1. The comparison of the results of the obtained recovery rate is as follows figure 1 B described.

[0079] According to the figure 1 B, it can be known that the optimal extraction solvent volume is 2mL.

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Abstract

The invention discloses an analysis and detection method for effectively identifying different species of Chinese soft-shelled turtle, comprising the following steps: taking the muscle tissue of the Chinese soft-shelled turtle to be tested and grinding it uniformly, then adding a dichloromethane-methanol mixed extraction solution as an extraction solution; After ultrasonic-assisted extraction, ultrapure water was added and then refrigerated and centrifuged; methylene chloride was added to the primary aqueous phase obtained by refrigerated centrifugation for secondary extraction, and the primary organic phase obtained by refrigerated centrifugation and the secondary organic phase obtained by secondary extraction were combined. , to obtain the extract; place the extract to dry under a nitrogen blower, make up to volume with acetonitrile, and pass through a 0.22 μm organic filter membrane as the liquid to be tested; analyze and detect the liquid to be tested by liquid chromatography-mass spectrometry, using m The phospholipid ions of / z790, 816, 818 and 866 were used as potential markers for species identification of Chinese soft-shelled turtle. This method can effectively distinguish the three Chinese soft-shelled turtles, Zhewu No.

Description

technical field [0001] The invention relates to the establishment of a lipidomics method for analyzing phospholipids in soft-shelled turtles, in particular, using this method to effectively identify different types of Chinese soft-shelled turtles and whether they are adulterated. Background technique [0002] The Chinese soft-shelled turtle is a high-value commercial species that is often consumed as a traditional Chinese medicine supplement and food in Asian countries such as China, Japan, and Korea. Chinese soft-shelled turtle contains nutritional factors such as protein, essential amino acids, polyunsaturated fatty acids, trace elements and other active substances, which can improve human immunity, promote metabolism, and prevent diabetes and anemia. In recent years, several new strains of soft-shelled turtles have been obtained through cross-breeding, which are expected to replace the current unimproved varieties. However, in the market, these high-value Chinese soft-sh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34
CPCG01N30/02G01N30/06G01N30/34G01N2030/062
Inventor 沈清俞喜娜
Owner ZHEJIANG GONGSHANG UNIVERSITY