CN116625772A - Micro-blood filtration method and application - Google Patents
Micro-blood filtration method and application Download PDFInfo
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- CN116625772A CN116625772A CN202310619799.1A CN202310619799A CN116625772A CN 116625772 A CN116625772 A CN 116625772A CN 202310619799 A CN202310619799 A CN 202310619799A CN 116625772 A CN116625772 A CN 116625772A
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- 238000001914 filtration Methods 0.000 title claims abstract description 73
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- 238000000034 method Methods 0.000 claims abstract description 36
- 239000011259 mixed solution Substances 0.000 claims abstract description 18
- 210000002966 serum Anatomy 0.000 claims abstract description 15
- 239000000644 isotonic solution Substances 0.000 claims abstract description 10
- 238000002156 mixing Methods 0.000 claims abstract description 9
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims abstract description 8
- 235000012041 food component Nutrition 0.000 claims abstract description 6
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- 229930003427 Vitamin E Natural products 0.000 claims abstract description 4
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims abstract description 4
- 235000019175 phylloquinone Nutrition 0.000 claims abstract description 4
- 239000011772 phylloquinone Substances 0.000 claims abstract description 4
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 claims abstract description 4
- MBWXNTAXLNYFJB-NKFFZRIASA-N phylloquinone Chemical compound C1=CC=C2C(=O)C(C/C=C(C)/CCC[C@H](C)CCC[C@H](C)CCCC(C)C)=C(C)C(=O)C2=C1 MBWXNTAXLNYFJB-NKFFZRIASA-N 0.000 claims abstract description 4
- 229960001898 phytomenadione Drugs 0.000 claims abstract description 4
- 235000019156 vitamin B Nutrition 0.000 claims abstract description 4
- 239000011720 vitamin B Substances 0.000 claims abstract description 4
- 235000019166 vitamin D Nutrition 0.000 claims abstract description 4
- 239000011710 vitamin D Substances 0.000 claims abstract description 4
- 150000003710 vitamin D derivatives Chemical class 0.000 claims abstract description 4
- 235000019165 vitamin E Nutrition 0.000 claims abstract description 4
- 229940046009 vitamin E Drugs 0.000 claims abstract description 4
- 239000011709 vitamin E Substances 0.000 claims abstract description 4
- 229940045997 vitamin a Drugs 0.000 claims abstract description 4
- 229940046008 vitamin d Drugs 0.000 claims abstract description 4
- 239000012528 membrane Substances 0.000 claims description 59
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 40
- 239000000243 solution Substances 0.000 claims description 38
- 239000011780 sodium chloride Substances 0.000 claims description 20
- 230000003204 osmotic effect Effects 0.000 claims description 13
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 12
- 238000001471 micro-filtration Methods 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 6
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 5
- 239000004677 Nylon Substances 0.000 claims description 4
- 239000004743 Polypropylene Substances 0.000 claims description 4
- 239000008156 Ringer's lactate solution Substances 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
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- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000004695 Polyether sulfone Substances 0.000 claims description 2
- 238000002615 hemofiltration Methods 0.000 claims description 2
- 229920003208 poly(ethylene sulfide) Polymers 0.000 claims description 2
- 229920006393 polyether sulfone Polymers 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 5
- 235000015097 nutrients Nutrition 0.000 abstract description 4
- 229920002521 macromolecule Polymers 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 abstract description 2
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 abstract description 2
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 abstract description 2
- 235000019155 vitamin A Nutrition 0.000 abstract description 2
- 239000011719 vitamin A Substances 0.000 abstract description 2
- 210000002381 plasma Anatomy 0.000 description 16
- 239000000706 filtrate Substances 0.000 description 12
- 238000001514 detection method Methods 0.000 description 9
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- 230000000052 comparative effect Effects 0.000 description 7
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- 229930003231 vitamin Natural products 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 101710154606 Hemagglutinin Proteins 0.000 description 3
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- 101710176177 Protein A56 Proteins 0.000 description 3
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- 239000000185 hemagglutinin Substances 0.000 description 3
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- 238000011068 loading method Methods 0.000 description 2
- -1 polytetrafluoroethylene Polymers 0.000 description 2
- 241001552669 Adonis annua Species 0.000 description 1
- 206010053567 Coagulopathies Diseases 0.000 description 1
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- 229930003270 Vitamin B Natural products 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004159 blood analysis Methods 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 238000009640 blood culture Methods 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
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- 230000036039 immunity Effects 0.000 description 1
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- 210000000265 leukocyte Anatomy 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 230000036285 pathological change Effects 0.000 description 1
- 229940021222 peritoneal dialysis isotonic solution Drugs 0.000 description 1
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- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D65/00—Accessories or auxiliary operations, in general, for separation processes or apparatus using semi-permeable membranes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/12—Composite membranes; Ultra-thin membranes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
- G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Water Supply & Treatment (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The application relates to the technical field of blood filtration, in particular to a micro-blood filtration method and application. The method comprises the following steps: (1) sucking a trace amount of blood; (2) Adding an isotonic solution into blood to dilute the blood, and uniformly mixing to obtain a mixed solution; (3) Squeezing and filtering the mixed solution for 8-18 s, and collecting filtered serum or plasma. The method is applied to detecting components in serum or plasma filtered by the method, wherein the components comprise nutritional components, and the nutritional components comprise at least one of vitamin A, vitamin D, vitamin E, vitamin K1 and B vitamins. The application can realize rapid filtration of trace blood, has higher separation rate, avoids the residue of macromolecular substances and harmful substances, and ensures better accuracy of detecting nutrient elements in blood.
Description
Technical Field
The application relates to the technical field of blood filtration, in particular to a micro-blood filtration method and application.
Background
The sample for blood examination mainly includes: whole blood, serum, and plasma. The whole blood after anticoagulation treatment is whole blood, and is mainly used for measuring blood cells, blood convention, blood sedimentation and the like. The pale yellow liquid obtained after removing blood cells is plasma, and is often used for measurement in aspects of coagulation, immunity and the like.
Traditional blood separation is typically performed using a centrifuge to determine the type and concentration of blood components (e.g., nutrients such as vitamins) from a plasma or serum sample obtained by centrifuging whole blood. However, centrifugation is laborious and takes a long time, and the upper serum or plasma is required to be carefully removed by using a pipette in the following steps, remixing is easy to cause, the possibility of hemolysis is increased, the extraction amount is small, and the centrifuge is required to be installed on a horizontal fixed site and cannot be used in a portable manner.
Referring to the patent application of publication No. CN109925884A, a method for whole blood filtration and a filter membrane structure for whole blood filtration are disclosed, which specifically comprise the following steps: (1) Selecting at least two layers of filtering membranes to form a filtering membrane structure in sequence from top to bottom, and treating the filtering membrane structure with hemagglutinin for later use; (2) Adding the whole blood sample into the filter membrane structure for filtering; (3) collecting the filtered serum or plasma. The filter membrane structure is formed by superposing at least two layers of filter membranes from top to bottom, the pore diameter of the superposed filter membranes from top to bottom is gradually reduced, and the area is gradually increased or equal. Through the filter membrane structure formed by superposing at least two layers of filter membranes, and the filter membrane structure is subjected to hemagglutinin treatment, so that erythrocytes in the whole blood sample are combined with hemagglutinin in the filter membrane and are trapped in the filter membrane structure, and therefore, the erythrocytes are ensured to be completely filtered and adsorbed in the filter membrane structure, and the plasma/serum can be effectively and stably separated from a very small amount of blood. However, the application selects to directly filter whole blood by using a double-layer membrane, and has the defects that the filter membrane needs to be pretreated before use, so that residues of treatment fluid in plasma can be possibly caused, the test result is influenced, and the whole operation time is long; the filter operation pressure is high, the speed is low, and the membrane hole blockage can be aggravated to a certain extent by the coagulant, so that the filter membrane structure needs to be frequently maintained or replaced, and the overall cost is high.
Disclosure of Invention
The application aims to provide a micro-blood filtering method which can realize the rapid filtration of micro-blood, has higher separation rate, avoids the residues of macromolecular substances and harmful substances and ensures that the detection accuracy of nutrient elements in blood is better.
A method of microfiltration comprising the steps of:
(1) Sucking trace blood with the trace amount of 1-200 uL;
(2) Adding an isotonic solution into the blood to dilute the blood, and uniformly mixing to obtain a mixed solution;
(3) Squeezing and filtering the mixed solution for 8-18 s, and collecting filtered serum or plasma.
By adopting the technical scheme, the isotonic solution refers to a solution with the osmotic pressure equal to that of plasma. In humans, normal plasma osmotic pressure is 280-320mmol/L, average 300mmol/L,1 mOsm/l=1 mmol/L, where solutions approximately equal to this osmotic pressure are isotonic solutions.
Preferably, the isotonic solution comprises at least one of sodium chloride solution, glucose solution, sodium lactate solution, sodium bicarbonate solution and sucrose solution.
Preferably, the isotonic concentration of the sodium chloride solution is 0.85% -0.93% w/v, and the osmotic pressure of the sodium chloride solution is 290-318mOsm/L; the isotonic concentration of the glucose solution is 5% -5.75% w/v, and the osmotic pressure of the glucose solution is 278-319mOsm/L; the isotonic concentration of the sodium lactate solution is 1.57% -1.87% w/v, namely the osmotic pressure is 285-334mOsm/L; the isotonic concentration of the sodium bicarbonate solution is 1.18% -1.42% w/v, and the osmotic pressure of the sodium bicarbonate solution is 283-338mOsm/L; the sucrose solution has an isotonic concentration of 9.6% to 10.9% w/v and an osmotic pressure of 280-319mOsm/L.
Preferably, in step (2), the isotonic solution is diluted 1 to 400 times with blood.
Preferably, in step (3), the extrusion pressure is 0-5MPa.
Preferably, in the step (3), the filtering is performed by a filtering membrane, wherein the filtering membrane is formed by combining a plurality of layers of membranes, and the number of layers of the plurality of layers of membranes is 2-4.
Preferably, the membrane material of the filtering membrane comprises at least one of GF, nylon, PES, PTFE, PP.
Preferably, the pore diameter of the membrane material is 0.22-10um.
Preferably, the pore diameter of the filtering membrane is gradually reduced from top to bottom, and the area of the filtering membrane is kept equal or gradually reduced from top to bottom.
It is a further object of the present application to provide a method for micro-hemofiltration, for detecting components in serum or plasma filtered by the method, said components comprising nutritional components comprising at least one of vitamin a, vitamin D, vitamin E, vitamin K1, B vitamins.
The beneficial effects of the application are as follows:
(1) The blood is diluted by the isotonic solution, preventing rapid clotting of the blood. Because blood can be coagulated when being exposed to air, when the blood drops into the filter membrane, the blood can be diffused due to capillary effect, a little time is needed in the middle, if the blood is coagulated too fast, and the blood is possibly solidified after the filtration is not completed due to the pressurization;
(2) The red blood cells and the white blood cells in whole blood are trapped by utilizing a membrane separation mode, so that the hemolysis phenomenon is avoided, and the recovery rate of plasma is high; the filtering operation is simple, and no pretreatment is needed;
(3) Instead of centrifugation, the method realizes the filtration of trace blood, shortens the blood pretreatment step, quickens the blood filtration rate, saves time, quickly realizes the filtration of trace blood, and has important application value for detecting vitamins in blood and certain diseases needing to be detected in time.
(4) Sample pretreatment steps of some instruments, such as liquid phase, mass spectrometry, etc., can be optimized; the membrane passing step before quantitative loading and reduction loading can be realized, and the centrifugation and membrane passing are further integrated;
(5) The operation is convenient, the cost is low, the application range is wide, and the method can be suitable for the pretreatment of samples measured by various instruments.
In conclusion, the application can realize the rapid filtration of trace blood, has higher separation rate, avoids the residue of macromolecular substances and harmful substances, and ensures that the detection accuracy of nutrient elements in blood is better.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of the microfiltration of blood of the present application.
Detailed Description
The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
Unless otherwise indicated, the materials or instruments used in the examples are readily available from commercial companies, wherein,
blood: whole blood
GF: glass fiber, nylon: nylon membrane, PES: polysulfone membrane, PTFE: polytetrafluoroethylene film, PP: and (3) polypropylene.
Example 1
Referring to fig. 1, a method of micro blood filtration comprises the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:6, and the blood is: sodium chloride solution = 1:6, uniformly mixing to obtain a mixed solution;
(3) Extruding the mixed solution under 15KPa pressure, filtering the mixed solution in a filtering membrane for 15s, wherein the filtering membrane is formed by combining multiple layers of membranes, the specific parameters of the filtering membrane are shown in table 1, and the filtered filtrate is B2.
Example 2
Referring to fig. 1, a method of micro blood filtration differs from example 1 in that the filtration time and specific parameters of the filtration membrane are different, and referring specifically to table 1, the filtrate is not B2.
Example 3
Referring to fig. 1, a method of micro blood filtration differs from example 1 in that the filtration time and specific parameters of the filtration membrane are different, and referring specifically to table 1, the filtrate is not B2.
Example 4
Referring to fig. 1, a method of micro blood filtration differs from example 1 in that the filtration time and specific parameters of the filtration membrane are different, and referring specifically to table 1, the filtrate is not B2.
Example 5
Referring to fig. 1, a method of micro blood filtration differs from example 1 in that the filtration time and specific parameters of the filtration membrane are different, and referring specifically to table 1, the filtrate is not B2.
Example 6
Referring to fig. 1, a method of micro blood filtration differs from example 1 in that the filtration time and specific parameters of the filtration membrane are different, and referring specifically to table 1, the filtrate is not B2.
Example 7
Referring to fig. 1, a method of micro blood filtration comprises the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:50, and the blood: sodium chloride solution = 1:50, uniformly mixing to obtain a mixed solution;
(3) Extruding the mixed solution under 15KPa pressure, filtering the mixed solution in a filtering membrane which is formed by combining a plurality of layers of membranes within 15s, collecting filtered filtrate, wherein the specific parameters of the filtering membrane are the same as those of the embodiment 1, and the filtrate is C2.
Example 8
Referring to fig. 1, a method of micro blood filtration comprises the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:400, and the blood is: sodium chloride solution = 1:400, uniformly mixing to obtain a mixed solution;
(3) Extruding the mixed solution under 15KPa pressure, filtering the mixed solution in a filtering membrane which is formed by combining a plurality of layers of membranes within 15s, collecting filtered filtrate, wherein the specific parameters of the filtering membrane are the same as those of the embodiment 1, and the filtrate is D2.
Comparative example 1
A method of microfiltration comprising the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) The blood is diluted without adding sodium chloride solution, and is centrifuged for 10min at a rotating speed of 3000r/min to obtain a centrifugal supernatant A1.
Comparative example 2
A method of microfiltration comprising the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) The blood is not diluted by adding sodium chloride solution, the blood is extruded under the pressure of 15KPa, the blood is pressed into a filter membrane for filtering, the filtering is completed within 15s, the filter membrane is formed by combining a plurality of layers of membranes, the specific parameters of the filter membrane are the same as those of the embodiment 1, and the filtered filtrate is collected, wherein the filtrate is A2.
Comparative example 3
A method of microfiltration comprising the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:6, and the blood is: sodium chloride solution = 1:6, uniformly mixing to obtain a mixed solution;
(3) The mixture was centrifuged at 3000r/min for 10min to obtain a supernatant B1.
Comparative example 4
A method of microfiltration comprising the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:50, and the blood: sodium chloride solution = 1:50, uniformly mixing to obtain a mixed solution;
(3) The mixture was centrifuged at 3000r/min for 10min to obtain a supernatant C1.
Comparative example 5
A method of microfiltration comprising the steps of:
(1) 100uL of blood is sucked, the blood is whole blood,
(2) Adding sodium chloride solution with isotonic concentration of 0.9% w/v into blood to dilute the blood, wherein the dilution ratio is 1:400, and the blood is: sodium chloride solution = 1:400, uniformly mixing to obtain a mixed solution;
(3) The mixture was centrifuged at 3000r/min for 10min to obtain a supernatant D1.
The application of the method according to micro blood filtration is applied to detecting components in serum or plasma filtered by the method, wherein the components comprise nutritional components, the nutritional components comprise at least one of vitamin A, vitamin D, vitamin E, vitamin K1 and vitamin B, and whole blood which is not filtered or centrifuged and is not diluted, A1, A2, B1, B2, C1, C2, D1 and D2 are respectively sent to a detection mechanism for detection, and the detection mechanism comprises: the detection method of the Hefeenoson medical test Co., ltd comprises the following steps: liquid chromatography tandem mass spectrometry, detection instrument: ABSCIEX Triple Quad TM 4500MD, comparative knotThe results are shown in Table 2.
TABLE 1
TABLE 2
Analysis of results:
(1) Referring to Table 1, it can be seen that the filtration time of the present application can be controlled to 10-15 s, which is much lower than the centrifugation time of comparative examples 1-5 by 10min;
(2) Referring to table 2, the detection of nutrients in blood found that:
a. the whole blood has a content of a part higher than A1 or A2 and a part lower than A1 or A2 because the whole blood is mainly composed of blood cells and plasma and has a high impurity content. At present, blood samples for clinical examination are mainly determined by adopting serum or plasma except whole blood for few projects such as blood cell examination, blood culture and the like, and other biochemical, immune and tumor markers and the like are detected, so that physiological components in the serum or plasma are relatively close to body tissue liquid, the physiological conditions of a body can be reflected more truly, and the pathological changes of the body are also more sensitive; there is also a reason that the blood is coagulated faster without dilution, which affects the accuracy of the detection result.
b. In a certain dilution multiple range (1-400 times), the content of the vitamin detection results of the centrifugal supernatant obtained by the centrifugal machine and the serum obtained by filtering is similar.
c. When the ratio of blood to sodium chloride is 1: 50-1:400, it can be seen from Table 2 that the higher vitamins have been detected to have reached the lower limit.
The preferred embodiments of the application disclosed above are intended only to assist in the explanation of the application. The preferred embodiments are not exhaustive or to limit the application to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the application and the practical application, to thereby enable others skilled in the art to best understand and utilize the application. The application is limited only by the claims and the full scope and equivalents thereof.
Claims (10)
1. A method of microfiltration of blood comprising the steps of:
(1) Sucking trace blood with the trace amount of 1-200 uL;
(2) Adding an isotonic solution into the blood to dilute the blood, and uniformly mixing to obtain a mixed solution;
(3) Squeezing and filtering the mixed solution for 8-18 s, and collecting filtered serum or plasma.
2. The method of micro blood filtration according to claim 1, wherein the isotonic solution comprises at least one of sodium chloride solution, glucose solution, sodium lactate solution, sodium bicarbonate solution, sucrose solution.
3. The method of microperforation according to claim 2 wherein the sodium chloride solution has an isotonic concentration of 0.85% to 0.93% w/v and an osmotic pressure of 290-318mOsm/L; the isotonic concentration of the glucose solution is 5% -5.75% w/v, and the osmotic pressure of the glucose solution is 278-319mOsm/L; the isotonic concentration of the sodium lactate solution is 1.57% -1.87% w/v, namely the osmotic pressure is 285-334mOsm/L; the isotonic concentration of the sodium bicarbonate solution is 1.18% -1.42% w/v, and the osmotic pressure of the sodium bicarbonate solution is 283-338mOsm/L; the sucrose solution has an isotonic concentration of 9.6% to 10.9% w/v and an osmotic pressure of 280-319mOsm/L.
4. The method of micropump blood filtration according to claim 1, wherein in step (2), the isotonic solution is diluted 1 to 400 times with blood.
5. The method of micro blood filtration according to claim 1, wherein in step (3), the pressing pressure is 0-5MPa.
6. The method according to claim 1, wherein in the step (3), the filtration is performed by using a filtration membrane comprising a combination of a plurality of layers, wherein the number of layers of the plurality of layers is 2 to 4.
7. The method of micro blood filtration of claim 6, wherein the membrane material of the filtration membrane comprises at least one of GF, nylon, PES, PTFE, PP.
8. The method of micro blood filtration according to claim 7, wherein the pore size of the membrane is 0.22-10um.
9. The method according to any one of claims 6 to 8, wherein the pore size of the filtration membrane is gradually reduced from top to bottom, and the membrane area of the filtration membrane is kept equal or gradually reduced from top to bottom.
10. Use of a method of micro-hemofiltration according to any one of claims 1 to 8 for detecting components in serum or plasma filtered by the method, said components comprising nutritional components comprising at least one of vitamin a, vitamin D, vitamin E, vitamin K1, B vitamins.
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