Method for measuring content of mixed amino acids

A mixed amino acid and determination method technology, applied in the field of determination of mixed amino acid content, can solve the problems of low amino acid sensitivity, low repeatability, low selectivity, etc., and achieve the effect of improving separation efficiency, high detection sensitivity, and high separation efficiency

Inactive Publication Date: 2016-12-07
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of the above deficiencies or improvement needs of the prior art, the present invention provides a method for determining th

Method used

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  • Method for measuring content of mixed amino acids
  • Method for measuring content of mixed amino acids
  • Method for measuring content of mixed amino acids

Examples

Experimental program
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Effect test

Embodiment 1

[0051] A method for determining the content of mixed amino acids, comprising the following steps:

[0052] (1) Perform pre-column derivatization on the samples and standards to be tested.

[0053] Using phenyl isothiocyanate for pre-column derivatization, the steps are as follows:

[0054] S1. Take 10 μl of the solution to be tested into a 1.5ml centrifuge tube, and dry it under vacuum at 65°C for 2 hours to obtain a dried sample;

[0055] S2. Dissolve the above-mentioned dried sample of S1 in 20 μl of methanol:water:triethylamine=2:2:1 solvent, and vacuum-dry at 65° C. for 30 minutes to obtain a pretreated sample;

[0056] S3. Take 20 μl of the derivatization reagent of methanol: water: triethylamine: phenyl isothiocyanate = 7:1:1:1, add it to the sample treated in S2 above, vortex, seal, and store at room temperature. Let it stand for 20 minutes, and then dry it under vacuum at 65°C for 30 minutes. The obtained samples are to be analyzed by high performance liquid phase, a...

Embodiment 2

[0073] A method for determining the content of mixed amino acids, comprising the following steps:

[0074] (1) Perform pre-column derivatization on the samples and standards to be tested.

[0075] Using phenyl isothiocyanate for pre-column derivatization, the steps are as follows:

[0076] S1. Take 10 μl of the solution to be tested into a 1.5ml centrifuge tube, and dry it under vacuum at 65°C for 2 hours to obtain a dried sample;

[0077] S2. Dissolve the above-mentioned dried sample of S1 in 20 μl of methanol:water:triethylamine=2:2:1 solvent, and vacuum-dry at 65° C. for 30 minutes to obtain a pretreated sample;

[0078] S3. Take 20 μl of the derivatization reagent of methanol: water: triethylamine: phenyl isothiocyanate = 7:1:1:1, add it to the sample treated in S2 above, vortex, seal, and store at room temperature. Let it stand for 20 minutes, and then dry it under vacuum at 65°C for 30 minutes. The obtained samples are to be analyzed by high performance liquid phase, a...

Embodiment 3

[0096] A method for determining the content of mixed amino acids, comprising the following steps:

[0097](1) Perform pre-column derivatization on the samples and standards to be tested.

[0098] Using phenyl isothiocyanate for pre-column derivatization, the steps are as follows:

[0099] S1. Take 10 μl of the solution to be tested into a 1.5ml centrifuge tube, and dry it under vacuum at 65°C for 2 hours to obtain a dried sample;

[0100] S2. Dissolve the above-mentioned dried sample of S1 in 20 μl of methanol:water:triethylamine=2:2:1 solvent, and vacuum-dry at 65° C. for 30 minutes to obtain a pretreated sample;

[0101] S3. Take 20 μl of the derivatization reagent of methanol: water: triethylamine: phenyl isothiocyanate = 7:1:1:1, add it to the sample treated in S2 above, vortex, seal, and store at room temperature. Let it stand for 20 minutes, and then dry it under vacuum at 65°C for 30 minutes. The obtained samples are to be analyzed by high performance liquid phase, an...

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Abstract

The invention discloses a method for measuring the content of mixed amino acids. The method is characterized by comprising the following steps: (1) performing pre-column derivatization treatment on a sample to be measured and a standard substance; (2) performing high performance liquid chromatography detection under the following conditions: a chromatographic column C18 being a high performance liquid reversed-phase chromatographic column, a flowing phase A being a formic acid aqueous solution with the concentration of 0.01 to 0.1 percent (v/v), a flowing phase B being prepared according to the ratio of acetonitrile to water of (70 to 30) to (50 to 50), an elution gradient being as follows: 0 to 12 min, with 90 percent of A and 10 percent of B; 12 to 20 min, with 70 percent of A and 30 percent of B, 20 to 30 min, with 52 percent of A and 48 percent of B; 30 min or more, with 100 percent of B, the column temperature being 30 to 50 DEG C, and detection UV being 254 nm; after the mixed amino acids are subjected to chromatographic separation according to the above mentioned conditions, performing mass spectrometric detection on the mixed sample for confirmation. The method has the advantages of high separation rate, high reproducibility, high detection sensitivity, high selectivity, quantification accuracy, simple operation, wider application range, suitability for detection of various free amino acids, and the like.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and more specifically relates to a method for determining the content of mixed amino acids. Background technique [0002] Amino acid analysis and detection are widely used, mainly in food, medical science, ecological environment and other fields. Specifically, foods rich in essential amino acids are considered to have high nutritional value, pathologically, changes in plasma amino acid and organic acid content are used as the main basis for the diagnosis and treatment of congenital metabolic diseases, the field of ecological environment, quantitative soil environment Changes in amino acid content and tracking of its transformation process were used as strong evidence to describe the N cycle. [0003] There are many methods to detect and determine the amino acid content. High-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method, as a new technology,...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02
Inventor 余龙江郑桂花周蓬蓬金文闻白云朱圆敏汪璨
Owner HUAZHONG UNIV OF SCI & TECH
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