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Method for detecting phospholipid in protein powder by aid of liquid chromatography-mass spectrometry technologies

A technology of technical detection and LC/MS, applied in the field of analytical chemistry, can solve the problems of difficult separation and accurate detection of phospholipids, complex components of protein powder matrix, etc.

Active Publication Date: 2018-11-27
BY HEALTH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, chromatography and mass spectrometry detection methods are the most accurate, because the matrix components of protein powder are complex, making it difficult to separate and accurately detect phospholipids in protein powder with existing technologies
Therefore, at present, there is no method for the quantitative detection of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI) in protein powder

Method used

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  • Method for detecting phospholipid in protein powder by aid of liquid chromatography-mass spectrometry technologies
  • Method for detecting phospholipid in protein powder by aid of liquid chromatography-mass spectrometry technologies
  • Method for detecting phospholipid in protein powder by aid of liquid chromatography-mass spectrometry technologies

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Embodiment 1

[0035] The embodiment of the present invention provides a specific method for detecting phosphatidylethanolamine by liquid chromatography-mass spectrometry, including:

[0036] Step 1. Accurately weigh 11.05 mg of the reference substance of phosphatidylethanolamine, place it in a 10ml volumetric flask, add chloroform:methanol (1:1 by volume) solution to dissolve and set the volume to the scale of the volumetric flask, shake well to obtain Reference substance stock solution (1.083mg / ml). The reference solution was stored in a refrigerator at -18°C in the dark for future use. Precisely pipette 1 ml of the control stock solution, dilute it with chloroform:methanol (1:1 by volume) solution and dilute it to a 25 ml volumetric flask to obtain the control intermediate solution. Precisely pipette 1ml of the contrast intermediate solution, dilute it with chloroform:methanol (volume ratio is 1:1) solution and settle to a 10ml volumetric flask to obtain the contrast test solution of pho...

Embodiment 2

[0078] The embodiment of the present invention provides a specific method for detecting phosphatidylinositol by liquid chromatography-mass spectrometry, including:

[0079] Step 1. Accurately weigh 23.68 mg of the reference substance of phosphatidylinositol (PI), place it in a 20ml volumetric flask, add chloroform:methanol (volume ratio is 1:1) solution to dissolve and settle to the scale of the volumetric flask, Shake well to obtain the reference substance stock solution (1.16mg / ml). The reference solution was stored in a refrigerator at -18°C in the dark for future use. Precisely pipette 1 ml of the control stock solution, dilute it with chloroform:methanol (1:1 by volume) solution and dilute it to a 25 ml volumetric flask to obtain the control intermediate solution. Precisely pipette 1ml of the control intermediate solution, dilute with chloroform: methanol (volume ratio is 1: 1) solution and settle to a 10ml volumetric flask to obtain the contrast test solution of phospha...

Embodiment 3

[0119] The embodiment of the present invention provides a specific method for detecting phosphatidylserine (PS) by liquid chromatography-mass spectrometry, including:

[0120] Step 1. Accurately weigh 20.04 mg of the reference substance of phosphatidylserine (PS), place it in a 20ml volumetric flask, add chloroform:methanol (volume ratio is 1:1) solution to dissolve and set the volume to the scale of the volumetric flask, shake Evenly, the reference substance stock solution (0.982mg / m1) was obtained. The reference solution was stored in a refrigerator at -18°C in the dark for future use. Precisely pipette 1 ml of the control stock solution, dilute it with chloroform:methanol (1:1 by volume) solution and dilute it to a 25 ml volumetric flask to obtain the control intermediate solution. Precisely pipette 1ml of the control intermediate solution, dilute with chloroform: methanol (volume ratio is 1: 1) solution and settle to a 10ml volumetric flask to obtain the contrast test sol...

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Abstract

The invention belongs to the technical field of analytical chemistry, and particularly relates to a method for detecting phospholipid in protein powder by the aid of liquid chromatography-mass spectrometry technologies. The method includes individually preparing gradient-concentration working standard solution of phosphatidyl ethanolamine (PE), phosphatidyl serine (PS) and phosphatidyl inositol (PI), detecting the working standard solution by the aid of the HPLC-MS / MS (high-performance liquid chromatography-mass spectrometry / mass spectrometry) technologies and plotting standard working curves;ultrasonically extracting, centrifuging and filtering to-be-detected samples by solvents to obtain test solution, detecting the test solution according to the HPLC-MS / MS technologies and substitutingresults into the standard working curves to obtain the concentration of the phosphatidyl ethanolamine, the concentration of the phosphatidyl serine and the concentration of the phosphatidyl inositolin the to-be-detected samples. The concentration is used as horizontal coordinates of the standard working curves, and peak areas are used as vertical coordinates of the standard working curves. HPLCsystem conditions include that C4 chromatographic columns or chromatographic columns with the performance identical to the performance of the C4 chromatographic columns are used as stationary phase. The method has the advantage that the technical shortcoming of deficiency of methods for quantitatively detecting the contents of PE, PS and PI in existing protein powder at present can be overcome bythe aid of the method.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a method for detecting phospholipids in protein powder using liquid mass spectrometry. Background technique [0002] Phospholipids, also known as phospholipids and phospholipids, refer to lipids containing phosphoric acid and belong to complex lipids. Phospholipids are the main components of biological membranes. They are divided into two categories: glycerophospholipids and sphingomyelins, which are composed of glycerol and sphingosine respectively. Phospholipids are amphiphilic molecules with a hydrophilic nitrogen or phosphorus head at one end and a long hydrophobic (oleophilic) hydrocarbon chain at the other end. For this reason, the hydrophilic ends of phospholipid molecules are close to each other, and the hydrophobic ends are close to each other. They often form lipid bilayers together with other molecules such as proteins, glycolipids, and choles...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N30/04G01N30/06G01N30/14
CPCG01N30/02G01N30/04G01N30/06G01N30/14G01N30/72G01N2030/047G01N2030/146
Inventor 叶少文苏昭仑李珍
Owner BY HEALTH CO LTD
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