Liquid chromatography mass spectrometry detection method of lactoferrin

A liquid chromatography mass spectrometry, lactoferrin technology, applied in the field of lactoferrin determination, can solve problems such as poor accuracy

Inactive Publication Date: 2018-08-14
北京市营养源研究所有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

The determination of lactoferrin concentration by spectrophotometry requires less equipment and can quickly determine the content of lactoferrin. It is suitable for rapid detection in the process of separation and purification in factories or for samples that do not require high accuracy of detection results. analysis, but its accuracy is relatively poor

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  • Liquid chromatography mass spectrometry detection method of lactoferrin
  • Liquid chromatography mass spectrometry detection method of lactoferrin
  • Liquid chromatography mass spectrometry detection method of lactoferrin

Examples

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Embodiment 1

[0066] The detection of lactoferrin in embodiment 1 milk powder

[0067] 1) Pretreatment of samples to be tested

[0068] Weigh 10 g of milk powder sample, ultrasonically dissolve it in a warm water bath at 35°C, and dilute it to 100 mL after cooling, as the milk powder sample solution.

[0069] Draw 200 μL of the above milk powder sample solution into a small test tube, add 180 μL of ammonium bicarbonate solution, mix well, add 15 μL of dithiothreitol solution, react at a constant temperature of 50°C for 30 minutes, take it out and cool it, add 45 μL of iodoacetamide solution, and stand in the dark 30min, add 10μL calcium chloride solution and 40μL trypsin solution, 37℃ constant temperature enzymatic hydrolysis for 4 hours, take it out, add 10μL formic acid, let stand at room temperature for 30min, and finally add appropriate amount of water to make the total volume of the enzymolysis solution 1.0mL, mix well After passing through a 0.22 μm filter membrane, the sample to be ...

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Abstract

The invention relates to a liquid chromatography mass spectrometry detection method of lactoferrin. The method comprises the following steps: respectively carrying out trypsin enzyme enzymolysis treatment on a to-be-detected sample and a lactoferrin standard substance to prepare to-be-detected sample probational liquid containing a specific peptide fragment and a lactoferrin standard working fluidtreatment test solution containing the specific peptide fragment; preparing specific peptide fragment standard working fluid by using a specific peptide fragment standard substance; carrying out highperformance liquid chromatography mass spectrometry detection. According to the liquid chromatography mass spectrometry determination method of the lactoferrin, disclosed by the invention, the content of non-denatured and thermally-denatured bovine lactoferrin can be determined by using a specific peptide fragment external standard method to quantify, adopting a lactoferrin standard and using enzymolysis to control an enzymolysis coefficient.

Description

technical field [0001] The invention relates to a method for measuring lactoferrin, belonging to the technical field of dairy product detection. Background technique [0002] Lactoferrin is a natural glycoprotein with immune function. It is transformed from transferrin. The crystal is red and consists of about 700 amino acid residues. Its molecular weight is 80ku and its isoelectric point is 8.7. It can be compared with Iron reversibly bound, mainly in milk, but also in tears, saliva, plasma, bile, pancreatic juice and neutrophils. The content of lactoferrin in human colostrum is relatively high, the concentration is 6-8 mg / mL; the concentration in cow colostrum is 1-2 mg / mL, and the content in normal milk is low, the concentration in human normal milk is 1-2 mg / mL, the concentration in cow milk is 0.02~0.35mg / mL. The content of lactoferrin in milk changes continuously at different stages of lactation. [0003] Lactoferrin is an active substance of the innate immune syst...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 白沙沙崔亚娟赵玥石明杨
Owner 北京市营养源研究所有限公司
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