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A method for simultaneous determination of active peptides and free amino acids

A technology for free amino acids and active peptides, applied in the field of analysis and detection, can solve the problem of not being able to measure active peptides and free amino acids at the same time

Active Publication Date: 2018-05-15
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The present invention aims to provide a method for simultaneous determination of active peptides and free amino acids to solve the problem in the prior art that amino acid analyzers cannot be used to accurately and quickly simultaneously determine the content of active peptides and free amino acids

Method used

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  • A method for simultaneous determination of active peptides and free amino acids
  • A method for simultaneous determination of active peptides and free amino acids
  • A method for simultaneous determination of active peptides and free amino acids

Examples

Experimental program
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Effect test

Embodiment 1

[0073] The analytical determination of embodiment 1 standard solution

[0074] (1) Preparation of elution buffer and ninhydrin derivative reagent

[0075] Prepare buffers B1-B5 according to Table 1; prepare ninhydrin derivatization reagents (ninhydrin reaction solution R1, reaction buffer R2, washing solution R3) according to Table 2.

[0076] The composition of table 1B1~B5 buffer solution

[0077]

[0078] The composition of table 2 ninhydrin derivative reagent

[0079]

[0080] (2) Amino acid and peptide standard solution preparation

[0081] Preparation of GSH (glutathione), Car (carnosine), P-Ser (phosphoserine), Tau (taurine), PEA (phosphoethanolamine), Urea (urea), Asp (aspartic acid), Hypro ( Hydroxyproline), Thr (threonine), Ser (serine), AspNH 2 (Asparagine), Glu (glutamic acid), GluNH2 (glutamine), Sar (sarcosine), a-AAA (α-aminocaproic acid), Pro (proline), Gly (glycine), Ala (alanine), Cit (citrulline), a-ABA (α-aminobutyric acid), Val (valine), Cys (cy...

Embodiment 2

[0093] The determination experiment of embodiment 2 linear range and detection limit

[0094] Dilute the standard mixed solution by a certain factor with 0.02mol / L hydrochloric acid solution, and use the optimized elution program in Table 3 for three gradient elution tests for each concentration solution.

[0095] By the retention time of going out peak as qualitative basis, make standard curve with the peak area of ​​measured chromatogram and its corresponding concentration, evaluate with linear correlation coefficient (R2); According to signal-to-noise ratio, when the peak of measured substance chromatographic peak When the height is 3 times of the noise (S / N=3), determine the minimum detection limit (LOD) of its mixed standard component; when the peak height of the chromatographic peak is 10 times of the noise (S / N=10), determine its mixed The limit of quantitation (LOQ) of a standard component is the lowest concentration value required to quantify the analyte.

[0096] Th...

Embodiment 3

[0101] The determination experiment of embodiment 3 recovery rate and precision

[0102] Take 2g shellfish sample tissue, add three mixed standard solutions with different concentrations of high, medium and low respectively to the sample, then add 15mL of 0.02mol / L dilute hydrochloric acid and 1mL of dithiothreitol with a concentration of 1wt%, and fully homogenize Ultrasonic cleaning was performed for 5 minutes, and then centrifuged with a refrigerated centrifuge (5000r, 4°C) for 10 minutes to collect the supernatant. Add 10 mL of dilute hydrochloric acid with a concentration of 0.02 mol / L to the remaining residue, stir, centrifuge again (5000 r, 4 °C) for 5 min, combine the supernatants, and dilute to 50 mL. After constant volume, pipette 2mL, add 2mL of 5wt% sulfosalicylic acid, centrifuge again (10000r, 4°C) for 10min, and then filter with a 0.22μm aqueous phase filter membrane to obtain the test solution.

[0103] Each concentration of the test solution was subjected to ...

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Abstract

The invention belongs to the field of analytical chemistry, which relates to a method for determining nutrients, and discloses a method for simultaneously determining active peptide and free amino acid. By adjusting the composition of a buffer, elution process and elution temperature during an amino acid analysis determinator standard determination process, a plurality of amino acid peaks and active peptide peaks are separated, and an amino acid automatic analyzer can perform one-step determination on the free amino acid and the active peptide, a method for preparing the amino acid and peptide simultaneously is established, and the disadvantages that the active peptide and a plurality of free amino acids cannot be separated, and the time consuming for testing is long by a traditional amino acid analyzer method are overcome.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a method for determining nutritional and flavor components, in particular to a method for simultaneously determining the content of active peptides and free amino acids. Background technique [0002] Free amino acids are an important evaluation index for the freshness and flavor of aquatic food, and are also an important part of non-protein nitrogen; different free amino acids have different tastes, presenting fresh, sweet, sour, bitter and other tastes, which will directly reflect the quality of aquatic products. The taste and freshness of the food. For example, threonine, serine, and proline have a sweet taste, while aspartic acid and glutamic acid play an important role in umami. Therefore, the content of free amino acids will be used as a potential indicator of important freshness for quality evaluation of many aquatic products. [0003] Active peptides refer to peptid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/8804G01N2030/8818
Inventor 邱伟强谢晶陈舜胜金银哲蓝蔚青桂娟
Owner SHANGHAI OCEAN UNIV
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