A kind of determination method of total phospholipid content in hemoglobin oxygen-carrying medicine

A technology of total phospholipid content and determination method, which is applied in the field of determination of total phospholipid content in hemoglobin oxygen-carrying drugs, can solve the problem of inability to quickly detect the total phospholipid content of hemoglobin oxygen-carrying drugs, complicated operations, and detection sensitivity that cannot meet the requirements, etc. problems, achieve good economic effects, improve detection throughput, and shorten sample detection time

Active Publication Date: 2018-02-23
BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Its disadvantage is that it cannot quickly detect the content of total phospholipids in hemoglobin oxygen-carrying drugs, and must test the content of a single phospholipid separately through multiple experiments, and finally add up the total phospholipid content
This operation is complicated, and the detection sensitivity cannot meet the requirements, so it is necessary to enrich the detection sample to a certain extent.

Method used

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  • A kind of determination method of total phospholipid content in hemoglobin oxygen-carrying medicine
  • A kind of determination method of total phospholipid content in hemoglobin oxygen-carrying medicine
  • A kind of determination method of total phospholipid content in hemoglobin oxygen-carrying medicine

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Experimental program
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Effect test

Embodiment 1

[0049] Preparation of malachite green solution: Precisely weigh 0.4 mg of malachite green, dissolve it in 100 ml of Pall ultrapure water, and prepare a malachite green solution with a weight content of 0.4%.

[0050] Preparation of ammonium molybdate solution: Accurately weigh 1.008 g of ammonium molybdate, dissolve it with 5 mol / L hydrochloric acid, and prepare ammonium molybdate solution with a weight content of 4.2%.

[0051] Preparation of Tween 20 solution: Accurately weigh 0.375 g of Tween 20, dissolve it in 25 ml of Pall ultrapure water, and prepare a Tween 20 solution with a weight content of 1.5%.

[0052] Preparation of the storage solution: the malachite green solution and the ammonium molybdate solution were mixed at a volume ratio of 3:1, filtered, and stored at room temperature in the dark for 2 to 3 weeks to obtain a storage solution.

[0053] Preparation of inorganic phosphorus chromogenic solution: mix the storage solution with the Tween 20 solution at a volum...

Embodiment 2

[0059] Sample to extract volume ratio selection test: At ordinary room temperature, when the sample to extract volume ratio is 1:4 and 1:8, it is easy to cause rapid protein denaturation of the hemoglobin solution sample, which may affect the extraction effect. Therefore, the prepared extract was pre-frozen in a deep-low temperature refrigerator for 30 minutes in advance.

[0060] When the volume ratio of sample to extract is 1:1, 1:2, 1:4, 1:8, in experiments with volume ratio of 1:1 and 1:2, the hemoglobin solution turns pink as the temperature returns to room temperature , the hemoglobin did not undergo severe polymerization and denaturation; while in experiments with volume ratios of 1:4 and 1:8, as the temperature returned to room temperature, the hemoglobin solution turned dark red, indicating that the hemoglobin was polymerized and denatured. The centrifugation operation was performed at room temperature, the speed of the centrifuge was 5000 rpm, and the centrifugation ...

Embodiment 3

[0062] Full-wavelength scanning and reaction kinetic curve of potassium dihydrogen phosphate standard solution: Measure 300 μl of potassium dihydrogen phosphate standard solution in a quartz cuvette, then add 100 μl of high-grade pure perchloric acid and 2ml of inorganic phosphorus chromogenic solution, and use Ultrospec6300pro UV The visible spectrophotometer scans the full wavelength from 200-900nm, the scanning wavelength interval is 1nm, and the scanning speed is 2649nm / min; from the beginning of adding the inorganic phosphorus chromogenic solution, a full-wavelength scanning is performed every 5 minutes, and the recording time is 0min, 5min, 10min, 15min, 20min. Measurement results such as figure 1 As shown, the reaction measurement wavelength can be used for determination at 560-690nm wavelength, and the maximum absorption peak corresponds to 634nm wavelength, so when using Ultrospec 6300pro ultraviolet-visible spectrophotometer to detect the test sample solution and pot...

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Abstract

The invention relates to a method for measuring total phospholipid content in hemoglobin oxygen-carrying drugs, which belongs to the technical field of medicine, and comprises the following steps: A. measurement operation; B. calculation of total phospholipid content in a sample. The method for determining the total phospholipid content in hemoglobin oxygen-carrying drugs replaces the existing HPLC method for determining the total phospholipid content in hemoglobin oxygen-carrying drugs by using an external standard method. Compared with the existing HPLC method, it improves the detection throughput. Quantity, shorten the sample detection time, reduce the detection cost, and reduce the operation steps, so as to realize the simple, fast and sensitive determination of the total phospholipid content in hemoglobin oxygen-carrying drugs.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for determining the content of total phospholipids in hemoglobin oxygen-carrying medicines. 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, which 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 cho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
Inventor 张学俊陈刚李燊周文涛蒋冰玉韩玎玎王红杨成民刘嘉馨
Owner BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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