CN115112463A - Micro blood filtering device and application thereof in blood nutrient element detection - Google Patents
Micro blood filtering device and application thereof in blood nutrient element detection Download PDFInfo
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- CN115112463A CN115112463A CN202210681811.7A CN202210681811A CN115112463A CN 115112463 A CN115112463 A CN 115112463A CN 202210681811 A CN202210681811 A CN 202210681811A CN 115112463 A CN115112463 A CN 115112463A
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- 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
- G01N1/34—Purifying; Cleaning
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
Abstract
The invention belongs to the technical field of biochemical separation, and discloses a micro blood filtering device and application thereof in blood nutrient element detection. The device can realize the filtration of trace blood, can quickly separate the whole blood in a short time, has high separation efficiency, has important application value for the detection of nutrient elements in the blood and certain diseases needing to be detected in time, has wide application range and has wide application prospect.
Description
Technical Field
The invention relates to the technical field of biochemical separation, in particular to a micro blood filtering device and application thereof in blood nutrient element detection.
Background
The nutrient elements are nutrient components which are necessary for maintaining normal life activities of a human body, mainly comprise carbohydrates, vitamins, mineral substances and the like, participate in various life activities of the human body, are related to certain diseases to a certain extent and reflect the health condition of the human body. Therefore, the method has great significance for the content measurement of various nutrient elements in human blood, human health detection, disease screening and the like.
There are various methods for detecting nutrient elements in the human body. For example, the detection method includes mass spectrometry, high performance liquid chromatography, electrochemical analysis, and the like. Among them, electrochemical analysis has made an important progress in the field of vitamin analysis because of its advantages such as high sensitivity, good selectivity, rapidness and economy. The principle of electrochemical vitamin detection is mainly based on that various vitamins generate oxidation-reduction reaction on the surface of a working electrode, so that qualitative and quantitative analysis is carried out on the vitamins. The components in human blood are complex and comprise proteins and amino acids; hormones, cholesterol, fatty acid, trace elements (potassium, iodine, zinc, copper, iron and magnesium), saccharides, enzymes, antigens, antibodies, ketones, alcohols, phenols, viruses, medicines, carbon dioxide, oxygen, water, urea, uric acid and the like, wherein most of the substances can also generate oxidation-reduction reaction on the surface of a working electrode, so that the interference on the detection of vitamins is generated, and therefore, the human blood sample is filtered, the interference of the substances on the reaction can be reduced, and the sensitivity and the accuracy of the detection are improved.
Hemofiltration (HF), which generally refers to the filtration of large amounts of liquid and solutes, i.e. ultrafiltrate, under the action of an ultrafiltration pressure by passing the blood through a filter in an extracorporeal circuit, by means of a machine (pump) or the patient's own blood pressure; at the same time, the electrolyte solution with similar components to the blood plasma, namely the replacement solution (replacement), is supplemented, thereby achieving the purpose of blood purification.
The samples for blood examination mainly include the following types: whole blood, serum and plasma. The whole blood after anticoagulation treatment is whole blood and is mainly used for measuring blood cells, blood routine, blood sedimentation and the like. The yellowish liquid obtained after removing blood cells is plasma, and is often used for measurement of blood coagulation, immunity, and the like.
Conventional filtration separation of blood is generally performed by using a centrifuge, and the kind and concentration of blood components (e.g., nutrients such as vitamins) are measured by centrifuging a plasma or serum sample obtained from whole blood. However, centrifugation is laborious, long, and requires careful removal of the supernatant serum or plasma using a pipette, which easily results in remixing, increases the possibility of hemolysis, requires a small amount of extraction, and the centrifuge needs to be installed in a horizontal stationary location and cannot be used in a portable manner.
Disclosure of Invention
In order to solve the above mentioned drawbacks in the background art, the present invention provides a micro blood filtration device and its application in blood nutrient detection, which can realize rapid filtration of micro blood, thereby improving separation rate and facilitating blood nutrient detection.
The purpose of the invention can be realized by the following technical scheme:
a micro blood filtering device comprises a dust cover, a filter membrane cover, a duplex tube and a reagent cup, wherein the reagent cup is sleeved outside one end of the duplex tube, the inner wall of the reagent cup is in threaded connection with the outer wall of the duplex tube, the filter membrane cover is sleeved outside the other end of the duplex tube, the filter membrane cover is clamped with the duplex tube, and the filter is arranged at the end part of the duplex tube close to one end of the filter membrane cover.
Preferably, one end of the duplex tube close to the reagent cup is a duplex-nozzle sample inlet tube, one end of the duplex tube close to the filter membrane cover is a single-connection sample dripping tube, and the duplex-nozzle sample inlet tube and the single-connection sample dripping tube are fixedly connected through a cylinder with a hollow structure.
Preferably, a quantitative capillary tube is arranged in the double-nozzle sample inlet tube, and the sampling volume of the double-nozzle sample inlet tube is 10-1000 muL.
Preferably, the filter setting is in the simply connected sample dropping pipe end portion, compresses tightly fixedly through the filter membrane lid to the filter, and the filter membrane lid middle corresponds simply connected sample dropping pipe department and has seted up out the liquid hole.
Preferably, the shape of the filter can be conical or round, and the filtering substrate connected inside the filter is a filter membrane which can effectively intercept macromolecular substances and harmful substances in blood.
Preferably, the material of the filter membrane is one of glass fiber, polysulfone membrane, polyesteramide, polypropylene, cellulose acetate membrane and mixed cellulose ester, the materials of the membranes can be hydrophilic and hydrophobic, and can also be used in combination, the plasma in the whole blood can be rapidly separated without hemolysis, the filter membrane is a single-layer or multi-layer combination, each layer of the multi-layer combination filter membrane can be different materials, and the thickness of the filter membrane is 0.05-20 mm.
The volume specification of the filtered blood is 10-1000 mu L, and the filtered blood plasma can be used for detecting vitamins, trace elements and other nutrient elements in the blood, thereby avoiding the residue of macromolecular substances and harmful substances and improving the sensitivity and accuracy of detection.
The invention has the beneficial effects that:
(1) the invention adds a filter and a filter membrane at the end of the double-pipe, and intercepts red blood cells and white blood cells in whole blood by a physical separation mode, thereby having no hemolysis and high plasma recovery rate. (2) The micro-blood filtering device can filter micro-blood, can quickly separate the whole blood in a short time, has high separation efficiency, has important application value for detecting nutrient elements in the blood and certain diseases needing to be detected in time, has wide application range and has wide application prospect.
Drawings
The invention will be further described with reference to the accompanying drawings.
FIG. 1 is an overall configuration of a hemodiafiltration apparatus according to the present invention;
FIG. 2 is a front view of a micro blood filtration device of the present invention;
FIG. 3 is a cross-sectional view A-A of a micro blood filtration device of the present invention;
FIG. 4 is a schematic diagram illustrating the use of the micro blood filtration device of the present invention;
FIG. 5 is a graph of CV before filtration of a whole blood sample in example 2 of the present invention;
FIG. 6 is a CV diagram of a whole blood sample after being filtered by a hemodiafiltration unit in accordance with example 2 of the present invention.
In the figure:
1-dust cap, 2-filter, 3-filter membrane cap, 4-duplex tube, 5-reagent cup, 21-filter membrane, 31-liquid outlet, 41-duplex sample inlet tube, 42-simplex sample dropping tube, 43-column and 44-holding cavity.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "opening," "upper," "lower," "thickness," "top," "middle," "length," "inner," "peripheral," and the like are used in an orientation or positional relationship merely to facilitate the description of the invention and to simplify the description, and are not intended to indicate or imply that the referenced components or elements must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered as limiting the present invention.
As shown in figures 1-3, a micro blood filtering device comprises a dust cover 1, a filter 2, a filter membrane cover 3, a duplex tube 4 and a reagent cup 5, wherein the reagent cup 5 is sleeved outside one end of the duplex tube 4, the inner wall of the reagent cup 5 is in threaded connection with the outer wall of the duplex tube 4, the filter membrane cover 3 is sleeved outside the other end of the duplex tube 4, the filter membrane cover 3 is clamped with the duplex tube 4, and the filter 2 is arranged at the end part of the duplex tube 4 close to one end of the filter membrane cover 3.
One end of the duplex tube 4 close to the reagent cup 5 is a duplex-mouth sample inlet tube 41, one end of the duplex tube 4 close to the filter membrane cover 3 is a single-joint sample dripping tube 42, and the duplex-mouth sample inlet tube 41 and the single-joint sample dripping tube 42 are fixedly connected through a cylinder 43 with a hollow structure.
The dual-nozzle sample inlet pipe 41 is internally provided with a quantitative capillary, and the sampling volume of the dual-nozzle sample inlet pipe 41 is 10-1000 muL.
The filter 2 is arranged at the end part of the single-connection sample dropping pipe 42, the filter 2 is compressed and fixed through the filter membrane cover 3, and the liquid outlet hole 31 is formed in the middle of the filter membrane cover 3 corresponding to the single-connection sample dropping pipe 42.
The shape of the filter 2 may be a cone or a circle, and the filtration substrate connected to the inside of the filter 2 is a filtration membrane 21.
The material of the filter membrane 21 is one of glass fiber, polysulfone membrane, polyester amide, polypropylene, cellulose acetate membrane and mixed cellulose ester, the filter membrane 21 is a single-layer or multi-layer combination, each layer of the multi-layer combination filter membrane 21 can be different materials, and the thickness of the filter membrane 21 is 0.05-20 mm.
As shown in FIG. 4, when blood filtration is performed by the device, the part of the filter membrane cover 3 is held by hand, a fixed amount (60 μ L) of sample is sucked through the double-nozzle sample inlet tube 41, the double-nozzle sample inlet tube 41 of the double-nozzle 4 is inserted into the reagent cup 5, wherein the sample diluent with a volume of 100-, the filtering of the blood is completed, the red blood cells and the white blood cells in the whole blood are intercepted by utilizing a physical separation mode, the hemolysis phenomenon is avoided, and the recovery rate of the plasma is high. The method has the advantages of thorough imbibition, high accuracy, capability of realizing the filtration of trace blood, capability of quickly separating the whole blood in a short time, high separation efficiency, capability of being used for detecting nutrient elements in the blood, important application value for certain diseases needing to be detected in time, wide application range and wide application prospect.
The volume specification of the filtered blood is 10-1000 mu L, and the filtered blood plasma can be used for detecting vitamins, trace elements and other nutrient elements in the blood, so that the residue of macromolecular substances and harmful substances is avoided, and the sensitivity and the accuracy of detection are improved. The specific application is as follows:
example 1
In the embodiment 1 of the invention, the micro blood filtering device is adopted to filter micro blood, and the liquid volume specification of the sample injection pipe with the duplex nozzle is 40-100 mu L.
Experimental groups: when filtering the whole blood sample, trace blood filter equipment advances the appearance through twin tube injection pipe 41, through filter 2 and filter membrane filtration back by going out liquid hole 31 and dripping into the reagent cup, can accelerate filter speed through extrusion reagent cup body during the filtration, whole blood filtration process can be accomplished in 20s, collects the plasma that the filtration obtained, then seals up and deposits in centrifuging tube, 4 ℃ preserves.
Control group: centrifuging the whole blood sample by a centrifuge at 3000r/min for 10min, standing for 5min, collecting supernatant, sealing in a centrifuge tube, and storing at 4 ℃.
And respectively sucking 200 mu L of samples from the experimental group, the control group and the whole blood sample by using a pipettor, respectively adding the samples into No. 1, No. 2 and No. 3 centrifuge tubes, standing for 5min, and then comparing.
The visual inspection result shows that the liquid in the centrifuge tube 1 is plasma obtained after the micro blood filtering device filters the liquid and is light yellow; the liquid in the centrifuge tube 2 is plasma obtained after centrifugation by a centrifuge and is light yellow; the liquid in the centrifuge tube 3 is an unprocessed whole blood sample, is darker in color, and has precipitates.
Through comparison of experimental results, the treatment effect of the trace blood filtering device on blood is basically consistent with the centrifugal effect of a centrifugal method on blood, red blood cells and white blood cells can be effectively removed, and light yellow plasma is obtained, which indicates that the treatment effect of the trace blood filtering device on blood can meet the requirement.
Example 2
The embodiment 2 of the invention provides a micro blood filtering device for detecting vitamin substances, which comprises the following steps:
(1) the electrochemical workstation was opened, 5ml of 0.1m pH 7PBS solution was added to the cell as an electrolyte, and the electrodes and platinum wire electrodes were placed on the cell platform and connected to the electrode clamps. Open the computer, open the CHI660e software, set the parameters, and wait for the test.
(2) And placing the ground glassy carbon electrode in an electrolytic cell to form a three-electrode system, and testing the electrochemical behavior of the three-electrode system by using a cyclic voltammetry method.
(3) The CV diagram is observed, after it is stabilized, the blank solution is tested with LSV and DPV, the electrochemical diagram is observed, and the test is repeated three times.
(4) To the electrolytic cell, 80. mu.L of a whole blood sample (unfiltered) was added, stirred for 1min, and then subjected to electrochemical detection with DPV, and the electrochemical pattern thereof was observed.
(5) The electrolytic cell was refilled with 80. mu.L of whole blood sample (processed through the hemodiafiltration unit), stirred for 1min, and then subjected to electrochemical detection with DPV, and the electrochemical patterns thereof were observed and compared for changes.
As can be seen from fig. 5, the current is unstable and relatively disordered, and the electrode surface is suspected to be polluted; fig. 6 shows that vitamin can be detected by analyzing the glassy carbon electrode modified by graphene oxide, and the peak value is obvious. Therefore, the micro blood filtering device can be compared to have a better application prospect in vitamin detection.
In the description herein, references to the description of "one embodiment," "an example," "a specific example," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed.
Claims (7)
1. The utility model provides a trace blood filter equipment, its characterized in that, includes shield (1), filter (2), filter membrane lid (3) dual tube (4) and reagent cup (5), reagent cup (5) are established to dual tube (4) one end outer lane cover, reagent cup (5) inner wall and dual tube (4) outer wall spiro union, filter membrane lid (3) are established to dual tube (4) other end outer lane cover, filter membrane lid (3) and dual tube (4) joint, dual tube (4) are close to filter membrane lid (3) one end tip and are equipped with filter (2).
2. The hemodiafiltration device according to claim 1, wherein the end of the duplex tube (4) close to the reagent cup (5) is a duplex-nozzle sample inlet tube (41), the end of the duplex tube (4) close to the filter membrane cover (3) is a single-joint sample dripping tube (42), and the duplex-nozzle sample inlet tube (41) and the single-joint sample dripping tube (42) are fixedly connected through a hollow cylinder (43).
3. The hemofiltration device according to claim 1, characterized in that a quantitative capillary tube is provided in the double-nozzle sample inlet tube (41), and the sampling volume of the double-nozzle sample inlet tube (41) is 10-1000 μ L.
4. The device for filtering trace blood according to claim 1, wherein the filter (2) is disposed at an end of the single-unit dripping tube (42), the filter (2) is pressed and fixed by the filter membrane cover (3), and the middle of the filter membrane cover (3) is provided with a liquid outlet hole (31) corresponding to the single-unit dripping tube (42).
5. A hemofiltration device according to claim 1, characterized in that the shape of the filter (2) can be conical or circular, the filtration matrix connected inside the filter (2) being a filter membrane (21).
6. The hemofiltration device according to claim 5, characterized in that the material of the filter membrane (21) is one of glass fiber, polysulfone membrane, polyesteramide, polypropylene, cellulose acetate membrane and mixed cellulose ester, the filter membrane (21) is a single layer or a multi-layer combination, each layer of the multi-layer combination filter membrane (21) can be different materials, and the thickness of the filter membrane (21) is 0.05-20 mm.
7. Use of a hemodiafiltration device according to any of claims 1 to 6 for the detection of blood nutrients, wherein the volume of the filtered blood is 10 to 1000 μ L, and the filtered plasma can be used for the detection of vitamins, trace elements and other nutrients in the blood, avoiding the residues of macromolecular substances and harmful substances, and improving the sensitivity and accuracy of the detection.
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| CN202210681811.7A CN115112463A (en) | 2022-06-15 | 2022-06-15 | Micro blood filtering device and application thereof in blood nutrient element detection |
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| CN202210681811.7A CN115112463A (en) | 2022-06-15 | 2022-06-15 | Micro blood filtering device and application thereof in blood nutrient element detection |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116625772A (en) * | 2023-05-25 | 2023-08-22 | 安徽九陆生物科技有限公司 | Micro-blood filtration method and application |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116625772A (en) * | 2023-05-25 | 2023-08-22 | 安徽九陆生物科技有限公司 | Micro-blood filtration method and application |
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