Diarrhetic shellfish poison standard sample as well as preparation method and application thereof

Inactive Publication Date: 2013-12-11
王秋艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is difficult to obtain standard samples of shellfish toxins. Firstly, because it is a highly toxic substance, countries around the world have strict control regulations on the entry and exit of shellfish toxins. In addition, there are many types of shellfish toxins, which are extremely unstable and difficult to preserve.
This will inevitably greatly limit the effective monitoring of shellfish poisoning, which will cause irreparable losses to shellfish farming enterprises
In my country, there is a blank for the preparation technology of standard samples of diarrheal shellfish toxins or quality control samples with traceable, certified and physical standards, and there is no report on the development of standard samples of diarrheal shellfish toxins

Method used

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  • Diarrhetic shellfish poison standard sample as well as preparation method and application thereof
  • Diarrhetic shellfish poison standard sample as well as preparation method and application thereof
  • Diarrhetic shellfish poison standard sample as well as preparation method and application thereof

Examples

Experimental program
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preparation example Construction

[0033] Such as image 3 Shown, the preparation method of diarrheal shellfish toxin standard sample of the present invention comprises the following steps: (A) sampling, impurity removal; (B) homogenate; (C) freeze-drying; (D) grinding, sieving; ( E) mixed sample, freeze-dried again; (F) package; (G) uniformity and stability test; (H) determine DSP content.

[0034] In a preferred embodiment, the preparation method of the diarrheal shellfish toxin standard sample of the present invention comprises the following steps:

[0035] (A) Take DSP-positive mussels, wash the shellfish shell thoroughly with water, cut off the adductor muscle, open the shell, rinse the interior with water to remove sediment and other foreign matter, take out the shellfish (to avoid damaging the midgut gland), and carefully cut out All midgut glands were spared.

[0036] (B) Add distilled water to the midgut gland sample, add methanol-acetone-ether mixed solution, homogenize and mix to obtain a homogenat...

Embodiment 1

[0058] Example 1: Take a DSP-positive mussel whose DSP content is 0.56 μg / g (fresh shellfish), wash the shell surface thoroughly with clean water, cut off the adductor muscle, open the shell, and rinse the interior with distilled water to remove sediment Separate the adductor muscle from the tissue connected to the glued part, remove all the shellfish, and carefully cut out all the midgut glands. Add an equal volume of distilled water and a mixed solution of methanol-acetone-ether (volume ratio 3:2:1) accounting for 1 / 10 of the volume of distilled water to the excised midgut gland, mix well and pre-freeze at -80°C for at least For 4 hours, use a vacuum freeze dryer (CHRiST Epsilon2-6D) to freeze-dry, then grind the tissue, pre-crumble, finely grind with a high-alumina ball mill, and pass through a 40-mesh sieve to obtain a DSP positive coarse powder. Add distilled water and methanol- The mixed solution of acetone-ether was stirred, mixed evenly, placed at -196°C to -20°C for 1...

Embodiment 6

[0065] Embodiment 6 Uniformity and Stability Inspection

[0066] 6.1 Homogeneity test of samples

[0067] Randomly select 15 bottles of samples prepared in Example 1, and do 2 repeatability tests for each bottle of samples. Use the enzyme-linked immunosorbent assay test in SN / T1996-2007 to analyze the DSP content of the standard sample, expressed in μg / g. All samples are tested in random order under repetitive conditions, that is, in the same laboratory by the same personnel using the same test methods and instruments for a short period of time.

[0068] The uniformity of the test results was evaluated by the single-factor multi-level analysis of variance method, and the following statistical formula was used for the single-factor multi-level analysis of variance. The results are shown in Table 2 and Table 3.

[0069] In order to test the uniformity of the samples, i samples are taken, and each sample is tested j times under repeated conditions.

[0070] Test average for ea...

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Abstract

The invention discloses a diarrhetic shellfish poison (DSP) standard sample as well as a preparation method and an application thereof. The method comprises the following steps: shelling fresh shellfish containing DSP masculine, taking out intestinal gland, adding organic solvent, homogenizing, freeze-drying primarily to obtain a coarse sample, grinding and screening the coarse sample to obtain freeze-dried powder, dispersing the freeze-dried powder in organic solvent again, and freeze-drying to obtain the DSP standard sample. The DSP standard sample has better uniformity and stability, can be used for proficiency testing activities in DSP testing projects in laboratories and for qualitative and quantitative detection of DSP, and can also be used for verification of detection methods, calibration of testing instruments and quality control and evaluation of testing results. The invention has the advantages that the raw materials cost is low, the preparation method is simple, and the obtained standard sample is a substantial standard sample, is uniform and stable, is easy to store, and is beneficial to the effective monitoring to shellfish poisons.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a shellfish toxin standard sample and its preparation method and application, in particular to a diarrhea shellfish toxin standard sample and its preparation method and application. Background technique [0002] Shellfish toxins are marine natural organic matter, and its formation is closely related to toxic algae blooms in the ocean. According to literature reports, there are about 184 to 267 species of microalgae that can form red tides, including 60 to 78 species of toxic microalgae. The toxins produced by toxic algae often enter the shellfish through the food chain, so these toxins are usually called shellfish poisoning, and people are poisoned by eating shellfish with shellfish poisoning. There are many types of shellfish poisons, the main ones found so far are paralytic shellfish poisons, diarrheatic shellfish poisons (DSP), neuropathic shellfish poisons (NSP) and amnesic shellf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D519/00C07D493/22G01N1/28G01N33/00
Inventor 王秋艳赵昕田苗程根武曹际娟
Owner 王秋艳
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