Method for detecting human lactoferrin content in human milk

Abstract

The invention discloses a method for detecting human lactoferrin content in human milk, using a high-pressure capillary electrophoresis analyzer as detection equipment. The method comprises the steps of: preparing a human lactoferrin standard solution to generate a standard curve between human lactoferrin content and analysis pattern integral quantities obtained by the high-pressure capillary electrophoresis analyzer; detecting human milk to be detected; and obtaining lactoferrin content in the human milk to be detected by using the standard cure according to the detection result of the human milk to be detected. The method provided by the invention has short required time and high precision, and a sample buffer solution with low toxicity and pungent smell and high sensibility is used.

Description

technical field

[0001] The invention relates to a method for detecting the content of human lactoferrin, in particular to a method for detecting the content of human lactoferrin in human milk, and belongs to the technical field of detection. Background technique

[0002] Human lactoferrin (Human lactoferrin, HLF) is a binding glycoprotein with a molecular weight of 76KDa. It has two carbohydrate side chains attached to its polypeptide chain. The carbohydrate content is about 7%. Its components include mannose, Galactose, N-acetylgalactosamine, fucose, sialic acid, etc. Human lactoferrin is a single polypeptide chain, the polypeptide chain is folded into "two leaves", each leaf structure contains an iron trivalent ion and a bicarbonate anion structure site, wherein the former exists in a deep crack, each Each molecule can bind two ferric ions. HLF contains a strong basic site near the N-terminal, so its isoelectric point is pH8.7. The content of human lactoferrin (HLF) in ...

Examples

Embodiment 1

[0025] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0026] (a) Dissolving the human lactoferrin standard substance in the sample buffer solution and preparing the standard solutions whose concentrations are respectively 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml and 0.25mg / ml, the human milk Put the ferritin standard solution in a centrifuge tube, shake it up, add it to the sample tube, clean it with ultrasonic waves, and use a high-pressure capillary electrophoresis analyzer for detection and analysis after shaking it up. 352612, 175234, 86524, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9974, the lowest detection limit is 0.1ug, the lowest The detected content is 0.01mg / ml, and the standard sample recovery ra...

Embodiment 2

[0033] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0034] (a) Dissolving the human lactoferrin standard substance in the sample buffer solution and preparing the standard solutions whose concentrations are respectively 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml and 0.25mg / ml, the human milk Put the ferritin standard solution in a centrifuge tube, shake it up, add it to the sample tube, clean it with ultrasonic waves, and use a high-pressure capillary electrophoresis analyzer for detection and analysis after shaking it up. 325687, 163518, 79685, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9997, the lowest detection limit is 0.002ug, the lowest The detected content is 0.01mg / ml, and the standard sample recovery ...

Embodiment 3

[0041] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0042] (a) the human lactoferrin standard substance is dissolved in the sample buffer solution and the preparation concentration is respectively the standard solution of 8mg / ml, 4mg / ml, 2mg / ml, 1mg / ml, 0.01mg / ml, the human lactoferrin Put the protein standard solution in a centrifuge tube, shake it up and add it to the sample tube, clean it with ultrasonic waves, shake it up and use a high-pressure capillary electrophoresis analyzer for detection and analysis. , 314385, 3124, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9969, the minimum detection limit is 2ng, the minimum detection The content is 0.01mg / ml, and the standard sample recovery rate is 92...

technical field

[0001] The invention relates to a method for detecting the content of human lactoferrin, in particular to a method for detecting the content of human lactoferrin in human milk, and belongs to the technical field of detection. Background technique

[0002] Human lactoferrin (Human lactoferrin, HLF) is a binding glycoprotein with a molecular weight of 76KDa. It has two carbohydrate side chains attached to its polypeptide chain. The carbohydrate content is about 7%. Its components include mannose, Galactose, N-acetylgalactosamine, fucose, sialic acid, etc. Human lactoferrin is a single polypeptide chain, the polypeptide chain is folded into "two leaves", each leaf structure contains an iron trivalent ion and a bicarbonate anion structure site, wherein the former exists in a deep crack, each Each molecule can bind two ferric ions. HLF contains a strong basic site near the N-terminal, so its isoelectric point is pH8.7. The content of human lactoferrin (HLF) in ...

Examples

Embodiment 1

[0025] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0026] (a) Dissolving the human lactoferrin standard substance in the sample buffer solution and preparing the standard solutions whose concentrations are respectively 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml and 0.25mg / ml, the human milk Put the ferritin standard solution in a centrifuge tube, shake it up, add it to the sample tube, clean it with ultrasonic waves, and use a high-pressure capillary electrophoresis analyzer for detection and analysis after shaking it up. 352612, 175234, 86524, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9974, the lowest detection limit is 0.1ug, the lowest The detected content is 0.01mg / ml, and the standard sample recovery ra...

Embodiment 2

[0033] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0034] (a) Dissolving the human lactoferrin standard substance in the sample buffer solution and preparing the standard solutions whose concentrations are respectively 4mg / ml, 2mg / ml, 1mg / ml, 0.5mg / ml and 0.25mg / ml, the human milk Put the ferritin standard solution in a centrifuge tube, shake it up, add it to the sample tube, clean it with ultrasonic waves, and use a high-pressure capillary electrophoresis analyzer for detection and analysis after shaking it up. 325687, 163518, 79685, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9997, the lowest detection limit is 0.002ug, the lowest The detected content is 0.01mg / ml, and the standard sample recovery ...

Embodiment 3

[0041] The method for detecting human lactoferrin content in human milk of the present invention comprises the following steps:

[0042] (a) the human lactoferrin standard substance is dissolved in the sample buffer solution and the preparation concentration is respectively the standard solution of 8mg / ml, 4mg / ml, 2mg / ml, 1mg / ml, 0.01mg / ml, the human lactoferrin Put the protein standard solution in a centrifuge tube, shake it up and add it to the sample tube, clean it with ultrasonic waves, shake it up and use a high-pressure capillary electrophoresis analyzer for detection and analysis. , 314385, 3124, according to the relationship between the concentration of the above standard solution and the corresponding peak area, use the software to draw the standard curve, the linear regression correlation coefficient of the standard curve obtained is 0.9969, the minimum detection limit is 2ng, the minimum detection The content is 0.01mg / ml, and the standard sample recovery rate is 92...