Determination method of lipid soluble saxitoxin in long-term-preserved oyster sample

A detection method, shellfish toxin technology, applied in the field of biological analysis, can solve the problem of long-chain fatty acid loss and other problems, and achieve the effect of data information support

Inactive Publication Date: 2017-08-08
徐静 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Alkaline methyl esterification is only suitable for fats; sulfuric acid-methanol methyl esterification is suitable for both free fatty acids and bound fatty acids (fats), but it is easy to cause loss of long-chain fatty acids (carbon chain length greater than 20 carbon atoms)

Method used

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  • Determination method of lipid soluble saxitoxin in long-term-preserved oyster sample
  • Determination method of lipid soluble saxitoxin in long-term-preserved oyster sample
  • Determination method of lipid soluble saxitoxin in long-term-preserved oyster sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Detection and analysis of fat-soluble shellfish toxins in long-term preserved oyster samples

[0050] 1. Sample pretreatment

[0051] The fresh oysters purchased in the Dalian market were removed from the shell and immediately put into the freezer at 0-4°C, -10°C and -20°C for frozen storage. During the test, the frozen sample was thawed at room temperature to be in a semi-frozen state. At this time, the shellfish meat was still in a frozen state. Remove the ice flakes attached to the outside of the oyster, wipe off the water with absorbent paper, and use a grinder in the sample processing room. Crush and weigh 10g of oyster meat as a sample.

[0052] 2. Prepare the sample total fat.

[0053] Take 10g of minced oyster meat sample, add 50mL of methylene chloride-methanol solution (V / V=2:1) ​​and 10ul of butylated hydroxytoluene (BHT) into a 100mL centrifuge tube (cover with holes), and then homogenize Centrifuge for 1min and 4000r / min for 5min; after taking...

Embodiment 2

[0090] Embodiment 2. Test result correction

[0091] According to the definition of mouse lethal dose, 1MU=4ug OA (paralytic shellfish poisoning)=6mg highly toxic FFA (free fatty acid), wherein highly toxic FFA is the sum of the three fatty acids described in the article, and this value will increase with the storage time It increases with the increase, and the value is known within seven days, see Table 2 for details.

[0092] Table 2

[0093] storage days 0 1 3 5 7 High toxic FFA content (ug / g) 83.80 103.27 111.10 111.99 144.47

[0094] According to the national standard method of paralytic shellfish toxin in mice, the initial injection volume of fresh shellfish meat is 1mL containing 20g shellfish meat. Assuming that the OA content in the initial shellfish meat is a ug / g, the virulence of 20g fresh shellfish meat (day 0) is: a*20 / 4(OA contribution to virulence)+83.3*20 / 6000(FFA contribution to virulence).

[0095] In order to eliminate the i...

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PUM

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Abstract

The invention discloses a determination method of lipid soluble saxitoxin in a long-term-preserved oyster sample, and the method comprises the following steps: (1) sample pretreatment; (2) extracting of free fatty acids in the sample; (3) boron trifluoride methyl esterification of the free fatty acids, and gas chromatography detection; and (4) result correction. The determination method uses a corrected boron trifluoride methyl esterification method for corrected detection of the lipid soluble saxitoxin the preserved oyster sample, and a method for obtaining the accurate content of toxins in an original production sample by detection of the saxitoxin in the preserved oyster sample is provided. Through effective correction, a real-time detection value can accurately reflect the original content of the saxitoxin in the preserved oyster sample, accurate data information can be obtained, and the retrospective study can be supported.

Description

technical field [0001] The invention belongs to the field of biological analysis, and relates to a method for detecting fat-soluble shellfish toxins in oysters, a complex biological sample preserved for a long time, and in particular to a method for correcting false positives in the detection results. Background technique [0002] Oysters are delicious, nutritious, and rich in unsaturated fatty acids, such as linoleic acid, linolenic acid, and docosahexaenoic acid. However, some experimental studies have found that unsaturated free fatty acids are not absolutely safe. As early as 1982, Japanese scholar Takagi T et al. used free fatty acid standards to inject mice intraperitoneally, and found that C20: 4n-6, C18: 3n-3, C20: 5n-3 showed high toxicity, while C18: 2n-6, C22:6n-3 showed low toxicity, and C18:1n-9 was almost non-toxic (Toxic effect of free polyenoic acids: a fat soluble marine toxin. Bull Fac Fish Hokkaido Univ 1982,33(4):255-262.). After that, Takagi T et al. f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/86
CPCG01N30/02G01N30/8665G01N2030/025
Inventor 徐静张琳邵筠乔曹际娟
Owner 徐静
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