Method for separation of palmitoyl pentaspeptide-3 by high performance liquid chromatography

A high-performance liquid chromatography, palmitoyl pentapeptide technology, applied in material separation, analysis materials, measuring devices and other directions, can solve problems such as large consumption, reduce pollution and improve separation and purification efficiency.

Inactive Publication Date: 2017-03-29
GL BIOCHEM SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a method for separating palmitoyl pentapeptide-3 by adopting high performance liquid chromatography, which mainly solves the need to consume a large amount of organic solvents such as acetonitrile when separating and purifying palmitoyl pentapeptide-3 by reverse phase chromatography

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  • Method for separation of palmitoyl pentaspeptide-3 by high performance liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. Sample treatment: add a little methanol to palmitoyl pentapeptide-3 and sonicate until the liquid is clear, filter through a filter membrane with a pore size of 0.45um and collect the filtrate for later use.

[0016] 2. Purification

[0017] Purification conditions: Chromatographic column: C4 chromatographic column, column diameter and length: 5cm×25cm. Mobile phase: Phase A: chromatographically pure acetonitrile; Phase B: 0.1% mass percent concentration of trifluoroacetic acid in methanol. Flow rate: 60-80ml / min. Detection wavelength: 230nm. Gradient: A%: 5%-70%, 30-50min. The injection volume is 3-5g.

[0018] Purification process: equilibrate the chromatographic column with 5% phase A and load the sample, the sample volume is 1-2.0L sample solution. Linear gradient elution, collect the target peak, and put the collected peptide solution in the collection bottle for later use.

[0019] 3. Salt conversion: Rinse the reversed-phase silica gel column with C4 as ...

Embodiment 2

[0022] 1. Sample treatment: add a little methanol to palmitoyl pentapeptide-3 and sonicate until the liquid is clear, filter through a filter membrane with a pore size of 0.45um and collect the filtrate for later use.

[0023] 2. Purification

[0024] Purification conditions: chromatographic column: C4 chromatographic column, column diameter and length: 5cm×25cm. Mobile phase: A phase: chromatographically pure acetonitrile; B phase: 0.1% mass percent trifluoroacetic acid ethanol solution. Flow rate: 60-80ml / min. Detection wavelength: 230nm. Gradient: A%: 5%-70%, 30-50min. The injection volume is 3-5g.

[0025] Purification process: equilibrate the chromatographic column with 5% phase A and load the sample, the sample volume is 1-2.0L sample solution. Linear gradient elution, collect the target peak, and put the collected peptide solution in the collection bottle for later use.

[0026] 3. Salt conversion: Rinse the reversed-phase silica gel column with C4 as the stationa...

Embodiment 3

[0029] 1. Sample treatment: add a little methanol to palmitoyl pentapeptide-3 and sonicate until the liquid is clear, filter through a filter membrane with a pore size of 0.45um and collect the filtrate for later use.

[0030] 2. Purification

[0031] Purification conditions: chromatographic column: C8 chromatographic column, column diameter and length: 10cm×25cm. Mobile phase: A phase: chromatographically pure acetonitrile; B phase: 0.1% mass percent trifluoroacetic acid methanol solution. Flow rate: 160-200ml / min. Detection wavelength: 230nm. Gradient: A%: 5%-70%, 30-50min. The injection volume is 10 -15g.

[0032] Purification process: equilibrate the chromatographic column with 5% phase A and load the sample, the sample volume is 3-5L sample solution. Linear gradient elution, collect the target peak, and put the collected peptide solution in the collection bottle for later use.

[0033] 3. Salt conversion: Rinse the reversed-phase silica gel column with C8 stationary...

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Abstract

The invention relates to a method for separation of peptide compounds and concretely relates to a method for separation of palmitoyl pentaspeptide-3 by high performance liquid chromatography. The method mainly solves the problem that in the palmitoyl pentapeptide-3 separation, because of small polarity, a lot of an organic solvent is consumed. The method comprises 1, preparing a chromatographic column used in the high performance liquid chromatography, wherein a silica gel column is used as a stationary phase, 2, selecting chromatographic grade acetonitrile as a mobile phase A, and a mobile phase B with a volume ratio of trifluoroacetic acid to methanol or ethanol of 0.001: 1, 3, selecting a mobile phase A gradient range of 5% to 70%, 4, selecting the detection wavelength of 230nm, and 5, selecting an acetate aqueous solution with a mass percent of 0.5% and chromatographic grade acetonitrile as salt transfer mobile phases. The method reduces the retention time of palmitoyl pentaspeptide-3 so as to improve the purification efficiency.

Description

technical field [0001] The invention relates to a method for separating peptide compounds, in particular to a method for separating palmitoyl pentapeptide-3 by high performance liquid chromatography. Background technique [0002] Palmitoyl pentapeptide-3, also known as pentapeptide, was first used by the dermatology medical field together with vitamin A and other anti-aging ingredients. It advertises that it can directly act on the dermis, promote collagen proliferation, and achieve skin firmness. Moisturizing ingredients work together to accelerate the firming and lifting effect. This peptide is a fragment of collagen, the most abundant protein in the skin. It is a pentapeptide composed of lysine, threonine, and serine. This pentapeptide is linked to the first amino acid through fat-soluble palmitic acid, and then the amino acids are linked in turn to form the peptide sequence Pal-Lys-Thr-Thr-Lys-Ser [Pal-KTTKS]. The reduction of collagen in the dermis of the skin is cons...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 秦敬国徐红岩
Owner GL BIOCHEM SHANGHAI
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