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High performance liquid chromatography method for measuring content of carnosine and anserine in fish head

A high-performance liquid chromatography and anserine technology, which is applied in the field of high-performance liquid chromatography for determining the content of carnosine and anserine in fish heads, can solve the problems of poor precision, cumbersome determination steps, low measurement efficiency, etc. The effect of small retention time deviation, good adsorption and separation effect, high precision and sensitivity

Active Publication Date: 2019-10-18
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to overcome the high cost of mass spectrometer detection when using high performance liquid chromatography and mass spectrometry to measure carnosine and anserine in organisms in the prior art, and most laboratories do not have detection conditions, which is not conducive to detection The method is widely used; if only high performance liquid chromatography is used for determination, the accuracy is poor, and the separation and purification requirements for samples are relatively high, complex pretreatment methods are required, the determination steps are cumbersome, and the measurement efficiency is low. Provide a High-performance liquid chromatography for the determination of carnosine and anserine content in fish heads. Appropriate fillers and measurement parameters are used in the determination, so that after simple pretreatment of fish head samples, carnosine and anserine in fish heads can be quickly and accurately determined. Anserine content

Method used

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  • High performance liquid chromatography method for measuring content of carnosine and anserine in fish head

Examples

Experimental program
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Effect test

Embodiment 1

[0040]Preparation of filler: Dissolve P123 and NaF in 1.5mol / L hydrochloric acid solution, stir at 40°C until completely dissolved; add cyanoethyltriethoxysilane dropwise to the above solution, stir for 25min and then dropwise Add ethyl orthosilicate, stir at 40°C for 7h; then add 1.5mol / L NaAlO 2 Hydrochloric acid solution, stirred at 40°C for 24h, NaF, P123, cyanoethyltriethoxysilane, ethyl orthosilicate, NaAlO in the solution 2 , The mass ratio of HCl is 1:175:42:520:30:2700; After adjusting the pH of the solution to 1.65 with ammonia water, move it into a Teflon-lined reactor, and place the reactor in a 150°C oven for activation for 40 hours; Take the reaction kettle out of the oven, adjust the pH of the solution to 6 with ammonia water after reaching room temperature, and then place it in an oven at 150°C for activation for 40 hours; after filtering, wash the product with distilled water and absolute ethanol, and dry it at 80°C for 10 hours , to obtain cyanated Al-SBA-15...

Embodiment 2

[0045] Preparation of filler: Dissolve P123 and NaF in 2mol / L hydrochloric acid solution, stir at 45°C until completely dissolved; add cyanoethyltriethoxysilane dropwise to the above solution, stir for 30min, then add dropwise Ethyl orthosilicate, stirred at 45°C for 6h; then add 2mol / L NaAlO 2 Hydrochloric acid solution, stirred at 45°C for 20h, NaF, P123, cyanoethyltriethoxysilane, ethyl orthosilicate, NaAlO in the solution 2 , The mass ratio of HCl is 1:180:45:550:50:2800; After adjusting the pH of the solution to 1.7 with ammonia water, move it into a Teflon-lined reactor, and place the reactor in a 200°C oven for activation for 36 hours; Take the reaction kettle out of the oven, adjust the pH of the solution to 6.5 with ammonia water after reaching room temperature, and then place it in an oven at 200°C for activation for 36 hours; after filtering, wash the product with distilled water and absolute ethanol, and dry it at 100°C for 8 hours to obtain cyanated Al-SBA-15; ad...

Embodiment 3

[0050] Preparation of filler: Dissolve P123 and NaF in 1mol / L hydrochloric acid solution, stir at 35°C until completely dissolved; add cyanoethyltriethoxysilane dropwise to the above solution, stir for 20min, then add dropwise Ethyl orthosilicate, stirred at 35°C for 8h; then add 1mol / L NaAlO 2 Hydrochloric acid solution, stirred at 35°C for 30h, NaF, P123, cyanoethyltriethoxysilane, ethyl orthosilicate, NaAlO in the solution 2 , The mass ratio of HCl is 1:170:40:500:20:2600; After adjusting the pH of the solution to 1.6 with ammonia water, move it into a Teflon-lined reactor, and place the reactor in a 100°C oven for activation for 48 hours; Take the reaction kettle out of the oven, adjust the pH of the solution to 5.5 with ammonia water after reaching room temperature, and then place it in an oven at 100°C for activation for 48 hours; after filtering, wash the product with distilled water and absolute ethanol, and dry it at 60°C for 12 hours , to obtain cyanated Al-SBA-15; ...

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Abstract

The invention relates to the technical field of a detection method, and discloses a high performance liquid chromatography method for measuring content of carnosine and anserine in a fish head. The high performance liquid chromatography method comprises the following steps of: taking soft tissues of the fish head, after mashing the soft tissues, adding extracting solution into the mashed soft tissues as a test sample, and carrying out heating extraction to obtain crude extract; after cooling the crude extract to the room temperature and carrying out refrigerated centrifugation, taking supernate and filtering the supernate with a millipore filter membrane; measuring the content of the carnosine and the anserine in the filtrate by high performance liquid chromatography, wherein mesoporous modified Al-SBA-15 loaded with copper ions is used as a filler in a chromatographic column of high performance liquid chromatography. According to the invention, the mesoporous modified Al-SBA-15 loadedwith the copper ions is used as the filler in the chromatographic column in high performance liquid chromatography analysis; the high performance liquid chromatography method has good absorption andseparation effects on the carnosine and the anserine, is high in detection accuracy and sensitivity and simple and convenient to operate; computer measurement can be carried out only by simple preprocessing without a fussy preprocessing step; meanwhile, deviations of a peak area and retention time are very small; and the high performance liquid chromatography method has very high accuracy.

Description

technical field [0001] The invention relates to the technical field of detection methods, in particular to a high-performance liquid chromatography method for determining the contents of carnosine and anserine in fish heads. Background technique [0002] Carnosine and anserine are functional dipeptides synthesized by L-histidine and β-alanine, which exist in the skeletal muscle and brain of animals, and are usually extracted from poultry skeletal muscle. Important physiological functions such as oxidation, anti-aging and neuromodulation. Therefore, it is necessary to establish a rapid and reliable detection method for carnosine and anserine. [0003] At present, there is no relevant national standard for the detection of carnosine and anserine, and the main determination methods are high performance liquid chromatography, ion exchange chromatography and capillary electrophoresis. However, the three methods have high requirements for the separation and purification of sampl...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 姜维李炜男严小军刘宇胡世伟李世杰李加亮
Owner ZHEJIANG OCEAN UNIV
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