Detection method of gizzerosine in fish meal

A technology for erostin and a detection method, which is applied to the field of high performance liquid chromatography detection of erostin in fish meal, can solve the problems of complex composition of fish meal, separation interference and the like, and achieves high accuracy, accurate calculation of sample content, Separation of highly practical effects

Active Publication Date: 2015-11-25
海南威尔检测技术有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Gyrogastric erosin is produced by the reaction of free histamine and free lysine or ε-NH2 of lysine in protein. The extraction method of this invention is to use 0.1mol / L HCl, mix and stir for 10 minutes, and only It can extract the free gizmosin in fish meal, but the gizmosin combined in protein cannot be extracted, and the composition of fish meal is very complex, and no purification step will interfere with the later separation

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  • Detection method of gizzerosine in fish meal
  • Detection method of gizzerosine in fish meal
  • Detection method of gizzerosine in fish meal

Examples

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Embodiment 1

[0036] A detection method for gizzard erosin in fishmeal, comprising the following steps:

[0037] (1) Sample pretreatment

[0038] Take 100mg of fish meal and add it to the hydrolysis tube, then add 5mL of 6mol / L hydrochloric acid, screw the cap tightly, put it in a drying oven at 109°C, hydrolyze for 22 hours, filter to obtain the hydrolyzate, and then adjust the pH value with 10mol / L sodium hydroxide solution To 6.8, after standing still, take the supernatant after centrifugation to obtain the crude extract;

[0039] (2) Enrichment and purification

[0040] Take 3mL methanol and 3mL ultrapure water to activate the MCX solid-phase extraction column, inject the crude extract obtained in step (1) into the MCX solid-phase extraction column for enrichment, and then rinse the MCX solid-phase extraction column with 3mL water and 3mL methanol respectively , and eluted with 3mL ammonia water-methanol solution after pumping dry, the eluate was blown dry with nitrogen at 40°C, added...

Embodiment 2

[0044] A detection method for gizzard erosin in fishmeal, comprising the following steps:

[0045] (1) Sample pretreatment

[0046] Take 500mg of fish meal and add it to the hydrolysis tube, then add 10mL of 6mol / L hydrochloric acid, screw the cap tightly, put it in a drying oven at 111°C, hydrolyze for 24 hours, filter to obtain the hydrolyzate, and then adjust the pH value with 5mol / L sodium hydroxide solution To 6.8, after standing still, take the supernatant after centrifugation to obtain the crude extract;

[0047] (2) Enrichment and purification

[0048]Take 3mL methanol and 3mL ultrapure water to activate the MCX solid-phase extraction column, inject the crude extract obtained in step (1) into the MCX solid-phase extraction column for enrichment, and then rinse the MCX solid-phase extraction column with 3mL water and 3mL methanol respectively , and eluted with 5mL ammonia water-methanol solution after pumping dry, the eluate was blown dry with nitrogen at 40°C, added ...

Embodiment 3

[0052] A detection method for gizzard erosin in fishmeal, comprising the following steps:

[0053] (1) Sample pretreatment

[0054] Take 300mg of fish meal and add it to the hydrolysis tube, then add 8mL of 6mol / L hydrochloric acid, screw the cap tightly, put it in a drying oven at 110°C, hydrolyze for 23 hours and filter to obtain the hydrolyzate, then adjust the pH value with 8mol / L sodium hydroxide solution to 7.00, after standing still, take the supernatant after centrifugation, and obtain the crude extract;

[0055] (2) Enrichment and purification

[0056] Take 3mL methanol and 3mL ultrapure water to activate the MCX solid-phase extraction column, inject the crude extract obtained in step (1) into the MCX solid-phase extraction column for enrichment, and then rinse the MCX solid-phase extraction column with 3mL water and 3mL methanol respectively , and eluted with 4mL ammonia water-methanol solution after pumping dry, and the eluate was blown dry with nitrogen at 40°C, ...

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Abstract

The invention discloses a detection method of gizzerosine in fish meal, and belongs to the technical field of detection of gizzerosine. The method disclosed by the invention comprises the following steps: sample pretreatment; enrichment and purification; on-line automated derivatization procedure. According to the detection method, gizzerosine combined in protein is extracted through sample pretreatment, and other purities and some acidic and neutral amino acid in fish meal are removed through enrichment and purification, so that separation is avoided from being disturbed, and the accuracy of detection of gizzerosine in fish meal is improved; besides, the sensitivity is high, the minimum detectable quantity is 1 mg / kg, and the detection method is suitable for detection of large-scale samples.

Description

technical field [0001] The invention relates to the technical field of detection of gizzard erosin, in particular to a high-performance liquid chromatography detection method for gizzard erosin in fish meal. Background technique [0002] Gizzerosine is formed by the free histidine in fish meal and its metabolite histamine and the protein in fish meal (ε-NH of lysine) in fish meal during heating and drying 2 ) reaction, and is mainly related to the temperature during fish meal processing. When the processing temperature exceeds 120°C, the content increases. The higher the temperature, the easier it is to produce toxins. It has been reported that 2.2mg / kg gizzardine can stimulate the secretion of gastric acid in poultry, resulting in "black vomiting". The content of gizzard erosive factor in normal fish meal is not more than 0.3mg / kg, its effect of promoting gastric acid secretion is 10 times that of histamine, and the ability of causing gizzard erosion is 300 times that of h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 赵艳刘耀敏张凤枰杨发树张艳红
Owner 海南威尔检测技术有限公司
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