Detection method of angiogenic peptide

An angiogenesis and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that the detection method of angiogenesis inhibitors has not yet been established

Pending Publication Date: 2021-07-06
BEIJING SAISHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] So far, a set of comprehensive, effective, rapid and stable dete

Method used

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  • Detection method of angiogenic peptide
  • Detection method of angiogenic peptide
  • Detection method of angiogenic peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] This embodiment provides an ultra-efficient liquid chromatographic analysis method of angiogenesis FPAT, which is as follows:

[0046] Test Instruments and Chromatography Conditions:

[0047] Waters H-CLASS Ultra High Performance Liquid Chromatography, Columns Bile silica gel as filler (2.1 × 100 mm, 1.7 μm, ACQUITY UPLC peptide beh, mixed with acetonitrile in 0.01 mol / L heptane sulfonate (pH value to 2.50) and acetonitrile to flow phase A, with acetonitrile to flow phase B, flow rate of 0.35 ml / min, the detection wavelength is 210 nm, the temperature is 60 ° C, the injection concentration is 1 mg / ml, the amount of injection is 0.8 ul, and the gradient elution is performed according to Table 1.

[0048] Table 1 Elution gradient

[0049]

[0050] Systematic Suitable Solution: Take LGX3, dissolve with water, and produce an LGX3 solution having a concentration of 0.1 mg / ml; Take the FPAT raw material, dissolved with LGX3 solution, and a systematic solution of 1 mg / ...

Embodiment 2

[0053] This embodiment provides an ultra-efficient liquid chromatographic analysis method of angiogenesis FPAT, which is as follows:

[0054] Test Instruments and Chromatography Conditions:

[0055] Waters H-Class Ultra Eco Liquid Chromatography, C18 Columns (2.1 × 100mm, 1.7 μm, ACQUITY UPLC PEPTIDE BEH, at 0.01 mol / L heptane sulfonate (pH value to 2.50) is a flow phase A, with acetonitrile to flow phase B, a flow rate of 0.35 ml / min, and the detection wavelength is 210 nm, temperature It was 60 ° C, the injection concentration was 1 mg / ml, and the amount of injection was 0.8 μL, and the gradient elution was followed.

[0056] Table 2 eluting gradient

[0057]

[0058] The FPAT raw material was dissolved in water into a sample solution of 1 mg / mL for injection. figure 2 As shown, it is seen from the figure that the main peak is quickly drawn, and there is no hanging column.

Embodiment 3

[0060] This embodiment provides an ultra-efficient liquid chromatographic analysis method of angiogenesis FPAT, which is as follows:

[0061] Test Instruments and Chromatography Conditions:

[0062] Waters H-Class Ultra Eco Liquid Chromatography, C18 Columns (2.1 × 100mm, 1.7 μm, ACQUITY UPLC PEPTIDE BEH, at 0.01 mol / L heptane sulfonate (pH value to 2.50) is a flow phase A, with acetonitrile to flow phase B, a flow rate of 0.35 ml / min, and the detection wavelength is 210 nm, temperature It was 60 ° C, the injection concentration was 1 mg / ml, and the amount of injection was 0.8 μL, and the gradient elution was followed.

[0063] Table 3 eluting gradient

[0064]

[0065] Determination of systematic solutions, image 3 As shown, the number of theoretical boards is calculated as 146510 in the FPAT peak; adjacent to the main peaks of post-surrogate LGX3 and the FPAT peak separation 1.60. However, the main peak of the raw material contains impurities and is severely trailing.

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Abstract

The invention provides a detection method of angiogenesis peptide, the detection method utilizes an ultra-high performance liquid chromatography analysis method to analyze an angiogenesis peptide sample, and the ultra-high performance liquid chromatography analysis condition comprises that sodium heptanesulfonate-acetonitrile is used as a mobile phase A, acetonitrile is used as a mobile phase B, and gradient elution is carried out. The invention provides the detection method of the angiogenesis peptide, the rapid and accurate detection of the angiogenesis peptide can be realized, compared with the traditional high performance liquid chromatography analysis method, the detection time can be saved by 75%, and the separation degree between the main peak and the front and back impurities, the number of theoretical plates and the trailing factor of the method can meet the method requirements.

Description

Technical field [0001] The present invention relates to the field of polypeptide detection, and more particularly to a method of detecting a blood vessel to form apex peptide. Background technique [0002] Vascular production apex peptide (FPAT) is a high-efficiency angiogenic inhibitor having a fatal or binding capacity of integrity, and the amino acid sequence C end containing an angiogenic endothelial suppressor is added to the arginine-glycine- Peptide sequence of aspartic acid. [0003] FPAT is a polypeptide fragment having a fifteen amino acids synthesized by chemical synthesis method, and specific structure asp-ser-gly-glu-gly-prop-pre-gly-glu-gly-gly-gly-val-arg · XC 2 Hide 4 O 2 , Molecular formula C 60 Hide 92 N 18 O 25 · XC 2 Hide 4 O 2 . In the process of chemical synthesis, many process impurities, such as isomer impurities, deletion peptides, insert peptides, fracture peptides, peptides, deprotoamide peptides, and amino acid side chains are not completely deprotecte...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/34G01N30/74G01N30/88
CPCG01N30/02G01N30/34G01N30/74G01N30/88G01N2030/8813G01N2030/8872
Inventor 苗苗吕晓利齐硕宋梦薇栾美丽滕宁宁
Owner BEIJING SAISHENG PHARMA
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