Fmoc amino acid analysis method

An analysis method and amino acid technology, applied in the field of Fmoc amino acid analysis, can solve problems such as the impact on the accuracy of analysis results, and achieve the effects of high repeatability, good stability, and low detection limit

Inactive Publication Date: 2019-01-01
南京肽业生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the prior art, high-performance liquid chromatography (HPLC) is mostly used for the analysis of amino acids, but different HPLC analysis conditions have a great impact on the accuracy of the analysis results of different amino acids
[0004] Arginine is an essential amino acid for the human body. In order to avoid by-products in the synthesis of peptides, the amino and guanidine groups of arginine must be protected; Fmoc-Arg(Pbf)-OH uses Fmoc(9-fluorenylmethyl Oxycarbonyl) to protect the amino group, use Pbf (2,2,4,6,7-pentamethylbenzofuran-5-sulfonyl) to protect the guanidine group; in the prior art, Fmoc-Arg (Pbf)-OH is not See the report

Method used

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  • Fmoc amino acid analysis method

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Effect test

Embodiment 1

[0021] An analysis method of Fmoc amino acid, described Fmoc amino acid is Fmoc-Arg (Pbf)-OH, and described analysis method comprises the following steps:

[0022] Step 1, Fmoc-Arg(Pbf)-OH is dissolved in acetonitrile, the concentration of the Fmoc-Arg(Pbf)-OH is 3mg / mL;

[0023] Step 2, the Fmoc-Arg(Pbf)-OH solution prepared in step 1 is used as a sample for high performance liquid chromatography analysis;

[0024] The chromatographic condition of described high performance liquid chromatography is:

[0025] Octadecylsilane-bonded silica gel is used as a filler, Sinochrom ODS-BP is used as a chromatographic column, gradient elution is adopted, the flow rate is 1.0ml / min, the column temperature is room temperature, the detection wavelength is 220nm, and the injection volume is 10 μL. Gradient elution is performed according to the following table:

[0026] time / min

[0027] The mobile phase A is a 0.1% trifluoroacetic acid solution; the mobile phase B is a 0.1% trif...

Embodiment 2

[0029] An analysis method of Fmoc amino acid, described Fmoc amino acid is Fmoc-Arg (Pbf)-OH, and described analysis method comprises the following steps:

[0030] Step 1, Fmoc-Arg(Pbf)-OH is dissolved in acetonitrile, the concentration of the Fmoc-Arg(Pbf)-OH is 2mg / mL;

[0031] Step 2, the Fmoc-Arg(Pbf)-OH solution prepared in step 1 is used as a sample for high performance liquid chromatography analysis;

[0032] The chromatographic condition of described high performance liquid chromatography is:

[0033] Octadecylsilane-bonded silica gel is used as a filler, Sinochrom ODS-BP is used as a chromatographic column, gradient elution is adopted, the flow rate is 2.0ml / min, the column temperature is room temperature, the detection wavelength is 220nm, and the injection volume is 10 μL. Gradient elution is performed according to the following table:

[0034] time / min

[0035] The mobile phase A is a 0.1% trifluoroacetic acid solution; the mobile phase B is a 0.1% trif...

Embodiment 3

[0037] An analysis method of Fmoc amino acid, described Fmoc amino acid is Fmoc-Arg (Pbf)-OH, and described analysis method comprises the following steps:

[0038] Step 1, Fmoc-Arg(Pbf)-OH is dissolved in acetonitrile, the concentration of the Fmoc-Arg(Pbf)-OH is 10mg / mL;

[0039] Step 2, the Fmoc-Arg(Pbf)-OH solution prepared in step 1 is used as a sample for high performance liquid chromatography analysis;

[0040] The chromatographic condition of described high performance liquid chromatography is:

[0041] Octadecylsilane-bonded silica gel is used as a filler, Sinochrom ODS-BP is used as a chromatographic column, gradient elution is adopted, the flow rate is 1.0ml / min, the column temperature is room temperature, the detection wavelength is 220nm, and the injection volume is 10 μL. Gradient elution is performed according to the following table:

[0042] time / min

[0043] The mobile phase A is a 0.1% trifluoroacetic acid solution; the mobile phase B is a 0.1% tri...

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Abstract

The invention belongs to the technical field of amino acid analysis, and particularly relates to an Fmoc amino acid analysis method, which comprises: 1, dissolving Fmoc-Arg(Pbf)-OH in acetonitrile, wherein the concentration of the Fmoc-Arg(Pbf)-OH is 2-10 mg / mL; and 2, carrying out high performance liquid chromatography analysis by using the Fmoc-Arg(Pbf)-OH solution prepared in the step 1 as a sample, wherein the chromatographic conditions of the high performance liquid chromatography are that octadecylsilane bonded silica gel is used as a filler, Sinochrom ODS-BP is used as a a chromatographic column, gradient elution is used, the flow rate is 1.0-2.0 ml / min, the column temperature is room temperature, and the detection wavelength is 220 nm. According to the present invention, with the method, the Fmoc amino acid Fmoc-Arg(Pbf)-OH can be precisely analyzed, and the method has advantages of high sensitivity, low detection limit, simple detection method, good stability and high repeatability.

Description

technical field [0001] The invention belongs to the technical field of amino acid analysis, in particular to a method for analyzing Fmoc amino acids. Background technique [0002] In the process of peptide synthesis, amino acids are prone to misconnection and many side reactions, and amino acids need to be protected. After the peptide is synthesized, the protecting group is removed, that is, fluorenylmethyloxycarbonyl (Fmoc) is a common protecting group. In the prior art, Fmoc is generally introduced by using Fmoc-Cl or Fmoc-Osu, and the purity of the synthesized Fmoc amino acid affects the product quality of the subsequent synthesized polypeptide. Therefore, the analysis of the purity of Fmoc amino acid is of great significance. [0003] In the prior art, high-performance liquid chromatography (HPLC) is mostly used for the analysis of amino acids, but different HPLC analysis conditions have a great impact on the accuracy of the analysis results of different amino acids. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 徐红岩贺秀慧杨明
Owner 南京肽业生物科技有限公司
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