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A hemoglobin and preservation method technology, applied in the field of hemoglobin preservation, can solve problems such as vasoconstriction, blood pressure increase, organ damage, etc.
Inactive Publication Date: 2015-09-02
TIANJIN POLYTECHNIC UNIV
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However, when these products are used in clinical practice, there are toxic and side effects such as
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Effect test
Embodiment 1
[0010] Embodiment 1 washing blood
[0011] Take an appropriate amount of human blood, add 3-6 times the volume of physiological saline (0.9% NaCl solution), and centrifuge at 4°C at 2800 rpm for 15-30 minutes. Get the supernatant containing white blood cells, plasma protein and platelets and squeeze it out, keep the lower layer of packed red blood cells, repeat the above washing process for 3-5 times, until the supernatant is basically colorless, in order to remove other substances other than red blood cells to the greatest extent. substances to obtain clean packed red blood cells.
Embodiment 2
[0012] Embodiment 2 pre-expansion
[0013] Take a certain amount of packed red blood cells and add an equal volume of sterilized high-purity deionized water for pre-expansion. At 24, 26, 28, and 30 minutes, observe the cell morphology at different pre-swelling times under a 400-fold microscope, and determine the best pre-swelling time according to the roundness of the cells.
Embodiment 3
[0014] Embodiment 3 hypotonic membrane rupture
[0015] Add 2 times the volume of water to the pre-swollen red blood cells to rupture the membrane, and the time is set at 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45 , 48, and 50 minutes, observe the cell morphology at different membrane rupture times under a 400-fold microscope, and determine the optimal membrane rupture time according to the degree of cell rupture, the concentration of purified hemoglobin, and the difficulty of filtering the membrane rupture solution. Then centrifuge at 10,000 rpm for 30-60 minutes, and the supernatant after centrifugation is a hemoglobin solution.
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Abstract
The invention relates to a hemoglobin storage method which is characterized by comprising the following steps of adding a proper quantity of human blood into normal saline (a 0.9% NaCl solution) with the volume being 3-6 times that of the human blood, centrifuging at the temperature of 4 DEG C and the rotating speed of 2800r/min for 15-30 minutes to obtain a supernatant containing leukocytes, plasma proteins and platelets, extruding the supernatant, retaining deposited erythrocytes at the lower layer, repeating the washing process for more than 3-5 times until the supernatant is basically colorless, and removing other substances except for the erythrocytes to the maximum extent to obtain clean deposited erythrocytes; adding a certain quantity of deposited erythrocytes into equivoluminal sterilized high-purity deionized water to pre-expand for 5-15 minutes, finally, adding water with the volume being 2 times that of the deposited erythrocytes to carry out low permeation for 10-20 minutes, centrifuging at the rotating speed of 10000r/min for 30-60 minutes, and extracting the supernatant after centrifuging to obtain a hemoglobin solution; and subpackaging the purified hemoglobin solution into a 10ml sample bottle, sealing by using paraffin, and storing the purified hemoglobin solution in a brown bottle filled with a pyrogallic acid solution at the temperature of 4 DEG C, wherein the content of hemoglobin does not exceed 3% after the sample is stored for 4 months and is kept on the relatively-low level.
Description
(1) Technical field: [0001] The invention relates to the field of biomedicine, in particular to a method for preserving hemoglobin. (two) background technology: [0002] The traditional blood transfusion method with transfusion of whole blood as the main means has played an extremely important role in medical rescue, but at the same time, it also has various shortcomings that are difficult to overcome. Risk of viral cross-infection, strained blood sources, short storage period and special facilities are required. In order to solve the above contradictions, people started research on red blood cell substitutes as early as the end of the 19th century. Red blood cell substitute refers to the general term for various oxygen carriers that can expand blood volume, maintain the balance of osmotic pressure inside and outside blood vessels, and have the function of carrying oxygen to replace human red blood cells. The development of blood substitutes has gone through the initial va...
Claims
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Application Information
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