CN113186643A - Preparation method of nanofiber-modified cellulose nitrate immunochromatographic membrane - Google Patents
Preparation method of nanofiber-modified cellulose nitrate immunochromatographic membrane Download PDFInfo
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- CN113186643A CN113186643A CN202110373906.8A CN202110373906A CN113186643A CN 113186643 A CN113186643 A CN 113186643A CN 202110373906 A CN202110373906 A CN 202110373906A CN 113186643 A CN113186643 A CN 113186643A
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- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical class O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 title claims abstract description 100
- 239000012528 membrane Substances 0.000 title claims abstract description 81
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 238000009987 spinning Methods 0.000 claims abstract description 96
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 79
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 79
- 239000002121 nanofiber Substances 0.000 claims abstract description 37
- 239000002798 polar solvent Substances 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 23
- 239000003960 organic solvent Substances 0.000 claims abstract description 15
- 238000003756 stirring Methods 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims description 55
- 238000010041 electrostatic spinning Methods 0.000 claims description 30
- 239000003495 polar organic solvent Substances 0.000 claims description 18
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical group CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 15
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/425—Cellulose series
- D04H1/4258—Regenerated cellulose series
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0069—Electro-spinning characterised by the electro-spinning apparatus characterised by the spinning section, e.g. capillary tube, protrusion or pin
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0092—Electro-spinning characterised by the electro-spinning apparatus characterised by the electrical field, e.g. combined with a magnetic fields, using biased or alternating fields
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
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- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4382—Stretched reticular film fibres; Composite fibres; Mixed fibres; Ultrafine fibres; Fibres for artificial leather
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Abstract
A preparation method of a nanofiber modified cellulose nitrate immunochromatographic membrane comprises the steps of adding cellulose nitrate, an organic solvent and a polar solvent into a container, and uniformly stirring to obtain a spinning solution; the commercially available NC membrane was carefully placed on the plate receiver, the instrument was turned on for spinning for 0-4h, and the spun membrane was dried. The immunochromatographic membrane is obtained by a simple one-step spinning method, and the immunochromatographic membrane with different porosities, protein adsorption amounts and flow rates is obtained by controlling the spinning time and regulating and controlling the spinning parameters and is used on an immunochromatographic test strip, so that the aims of improving the sensitivity, specificity and repeatability of detecting substances such as protein, DNA and the like are fulfilled.
Description
Technical Field
The invention belongs to the technical field of material science, and particularly relates to a preparation method of a nanofiber modified cellulose nitrate immunochromatographic membrane.
Background
The Lateral Flow Assay (LFA) test strip has small volume, convenient carrying and short detection time, the used drug reagent is stable and easy to store, large instruments and equipment are not needed in qualitative detection, and semi-quantitative or quantitative detection can be carried out through the detection of a simple reading analysis instrument, so the LFA test strip is an analysis means which is very suitable for medical diagnosis, and has wide application in a plurality of fields of food safety, water resource, medical safety, life health and the like. At present, several common LFA strips, such as ovulation test strips, pregnancy test strips, urine test strips, and the like, are sold in hospitals and pharmacies and become successful cases for commercialization of LFA. However, LFAs have limited sensitivity and remain challenging in low concentration detection applications.
In order to improve the sensitivity of LFA, researchers have adopted many methods, including sample pretreatment, modification of labeled antibody nanoparticles, nanoenzyme signal enhancement, and targeted immobilization of antibodies, or introducing chemical functional groups on the surface of test paper, increasing the specific surface area of the test paper, modifying the key component, namely, a Nitrocellulose (NC) membrane, etc. However, these methods of increasing the sensitivity of LFAs require additional equipment, complicated operations or specialized personnel, are often time consuming, and are not suitable for field testing.
Disclosure of Invention
The invention aims to provide a preparation method of a nanofiber modified nitrocellulose immunochromatographic membrane.
In order to achieve the above object, the technology adopted by the present invention is as follows:
a preparation method of a nanofiber modified cellulose nitrate immunochromatographic membrane comprises the following steps:
step 1, accurately weighing nitrocellulose according to the total required amount of a spinning solution, wherein the mass percent of nitrocellulose serving as a solute in the spinning solution is 15-40%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5:1-1: 5;
step 3, preparing a spinning solution: adding cellulose nitrate, an organic solvent and a polar solvent into a container, and uniformly stirring to obtain a light yellow transparent clear spinning solution;
step 4, electrostatic spinning operation: a commercially available Nitrocellulose (NC) membrane was carefully placed on a plate receiver, and the spinning solution was sonicated to remove air bubbles from the solution; then injecting the solution into an injector and then installing the injector on an electrostatic spinning machine, setting spinning voltage, spinning solution injection speed and the angle between a needle head and a flat plate, starting an instrument for spinning, and enabling the spinning nanofibers to be uniformly distributed and covered on the surface of the NC membrane;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ to obtain the NC film modified by the nano fibers.
The organic solvent is acetone.
The polar solvent is N, N-dimethylformamide.
The spinning voltage is 10-30 KV; the injection speed of the spinning solution is 0.1-3 mL/h; the diameter of the spray head is 0.1-1 mm; the moving speed is 10-40mm/s, and the reciprocating distance is 180-240 mm.
The drying temperature of the invention is not more than 40 ℃ and the drying time is 4-24 h.
Compared with the prior art, the invention has the beneficial effects that:
the electrostatic spinning method is a simple and economical method for preparing the nanofiber modified cellulose nitrate immunochromatographic membrane. The method can obtain the membrane material with uniform fiber appearance and uniform thickness, a layer of NC fiber is electrostatically spun on the surface of the commercially available NC membrane, and the electrostatic spinning time is controlled, so that the porosity of the surface of the NC fiber is adjusted, the adsorption performance of protein is improved, the flow rate is reduced, the antigen-antibody reaction time is prolonged, and the aim of improving the sensitivity is finally fulfilled.
Drawings
FIG. 1 is a scanning electron microscope image of a comparison of a commercially available NC membrane according to a method for preparing a nanofiber-modified nitrocellulose immunochromatographic membrane of the present invention;
FIG. 2 is a scanning electron microscope image of an immunochromatographic membrane prepared by a preparation method of a nanofiber-modified cellulose nitrate immunochromatographic membrane of the present invention;
FIG. 3 is a graph showing the contact angle of a video optical contact angle measuring instrument of a commercially available NC membrane in comparison with a method for preparing a nanofiber-modified nitrocellulose immunochromatographic membrane;
FIG. 4 is a contact angle diagram measured by a video optical contact angle measuring instrument of an immunochromatographic membrane prepared by a preparation method of a nanofiber-modified cellulose nitrate immunochromatographic membrane of the present invention;
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention prepares the immunochromatographic membrane which can be used as a carrier T-line and C-line on a test strip by an electrostatic spinning technology.
The invention relates to a nanofiber modified cellulose nitrate immunochromatographic membrane prepared by a simple one-step electrostatic spinning method, which is used for obtaining immunochromatographic membranes with different porosities, protein adsorption amounts and flow rates by controlling spinning time and regulating and controlling spinning parameters, and is used on an immunochromatographic test strip, so that the purposes of improving the sensitivity, specificity and repeatability of detecting substances such as protein, DNA and the like are achieved.
The high-volatility polar organic solvent is a ketone organic solvent, and the high-conductivity polar solvent is an amide organic solvent.
The invention prepares a nanofiber modified cellulose nitrate immunochromatographic membrane by a simple one-step electrostatic spinning method, adjusts spinning time and spinning parameters in the spinning process, and the like, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 15-40%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5:1-1: 5;
step 3, preparing a spinning solution: adding cellulose nitrate, a high-volatility polar organic solvent and a high-conductivity polar solvent into a container, and stirring at room temperature for 24 hours (the stirring speed is not limited) to obtain a light yellow transparent clear spinning solution;
step 4, electrostatic spinning operation: the commercially available NC membrane was carefully placed on a flat plate receiver and the spinning solution was sonicated in a sonicator to remove air bubbles from the solution. Then injecting into an injector, installing a needle head and a pipe, finally installing on an electrostatic spinning machine, setting spinning voltage, spinning solution injection speed, rotating speed of a receiving fiber flat plate and angle between the needle head and the flat plate, starting an instrument for spinning and enabling spinning nano fibers to be uniformly distributed and covered on the surface of an NC membrane;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ to obtain the NC film modified by the nano fibers.
The invention is characterized in that the cellulose nitrate has the molecular formula [ C6H7O2(ONO2)a(OH)3-a]n。
Wherein n is the degree of polymerization and a is the degree of esterification, is a high molecular weight polymer. The higher the molecular weight is, the more entanglement of the molecular chain in the solution is increased, and the jet flow is stretched in the electrostatic spinning process, so that the molecular chain is well oriented, the action of external force is resisted, the spinning breakage is avoided, and the nano fibers are easily and uniformly collected on a flat receiver.
The invention is characterized in that the high-volatility polar organic solvent is acetone.
The high-volatility polar organic solvent in the step 2 is very volatile in the spinning process, the solvent plays a role in unfolding polymer molecular chains in the spinning process, the solution forms jet flow, the jet flow is highly stretched by electromagnetic force, the polymer molecular chains are reoriented and arranged, and the jet flow is solidified into polymer fibers along with the volatilization of the solvent. The volatility of the solvent can affect the stretching and curing of the jet. The selection of a highly volatile polar organic solvent can well yield uniform fibers collected on a flat plate.
The invention is characterized in that the high-conductivity polar solvent is N, N-dimethylformamide.
The high-conductivity polar solvent in the step 2 is a solvent with a higher dielectric constant, and the size of the diameter of the fiber is directly influenced by the dielectric constant of the solvent, generally, the higher the dielectric constant is, the smaller the diameter is, and the diameter distribution is narrower.
The stirring time in step 3 was 24 h. The method is favorable for forming uniform, transparent and viscous spinning solution and is favorable for spinning; the stirring temperature is room temperature, and the stirring speed is ensured to be not splashed by liquid. All conditions for preparing the spinning solution are not critical.
The ultrasonic time in the step 4 is 0.5h until bubbles are removed, and the spinning time is 0-4 h; the spinning voltage is 10-30 KV; the injection speed of the spinning solution is 0.1-3 mL/h; the diameter of the spray head is 0.1-1 mm; plate parameters: the moving speed is 10-40mm/s, and the reciprocating distance is 180-240 mm.
In the step 4, the temperature of the vacuum drying oven does not exceed 40 ℃, the drying time is different from 4 to 24 hours (according to the thickness of the spinning membrane), and then a clean glove is worn when the spun fiber membrane is taken, so that the cleanness and the purity of the NC membrane are ensured.
The immunochromatographic membrane obtained by the electrostatic spinning method with simple operation is shown in fig. 1 and 2, wherein fig. 1 is a surface topography of a commercially available NC membrane, and fig. 2 is a surface topography of the immunochromatographic membrane spun by the present invention. Comparing the two figures, the appearance in fig. 1 is uniform beaded, large pores, dense beads; unlike fig. 1, the morphology in fig. 2 is fibrous, with uniform fiber diameter and larger pores. Fig. 3 and 4 are water contact angle graphs of a commercially available NC membrane and an immunochromatographic membrane of the present invention, respectively, in which fig. 3 shows a contact angle of 0 ° and is a hydrophilic membrane, and fig. 4 shows a contact angle of 117 ° and is a hydrophobic membrane.
Example 1
The immunochromatographic membrane is obtained by an electrostatic spinning method with simple operation, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 18%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 1: 1;
step 3, preparing a spinning solution: respectively adding cellulose nitrate, a high-volatility polar organic solvent and a high-conductivity polar solvent into a small beaker, and stirring for 24 hours at room temperature to obtain a light yellow transparent clear solution to be used for spinning;
step 4, electrostatic spinning operation: the solution was sonicated in a sonicator for 0.5h until the bubbles in the liquid were completely removed. Carefully placing a commercially available NC membrane on a flat plate receiver, then injecting a spinning solution into a 10mL syringe, installing a needle and a pipe, finally installing the needle and the pipe on an electrostatic spinning machine, starting an instrument to spin, uniformly distributing and covering spinning nano fibers on the surface of the NC membrane, and spinning for 10 min; the spinning voltage is 15 KV; the injection speed of the spinning solution is 0.25 mL/h; the diameter of the spray head is 0.45 mm; the moving speed is 20mm/s, and the reciprocating distance is 190 mm;
and 5, after spinning is finished, drying the NC membrane with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ for 3h to obtain the NC membrane modified by the nano fibers.
Example 2
The immunochromatographic membrane is obtained by an electrostatic spinning method with simple operation, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 22%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5: 4;
step 3, preparing a spinning solution: respectively putting cellulose nitrate, a high-volatility polar organic solvent and a high-conductivity polar solvent into a small beaker, and stirring for 24 hours at room temperature to obtain a light yellow transparent clear solution to be used for spinning;
step 4, electrostatic spinning operation: the solution was sonicated in a sonicator for 0.5h until the bubbles in the liquid were completely removed. Carefully placing a commercially available NC membrane on a flat plate receiver, then injecting a spinning solution into a 10mL syringe, installing a needle and a pipe, finally installing the needle and the pipe on an electrostatic spinning machine, starting an instrument to spin, uniformly distributing and covering spinning nano fibers on the surface of the NC membrane, and spinning for 20 min; the spinning voltage is 18 KV; the injection speed of the spinning solution is 1.0 mL/h; the diameter of the spray head is 0.6 mm; the moving speed is 25mm/s, and the reciprocating distance is 200 mm;
and 5, after spinning is finished, drying the NC membrane with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ for 5 hours to obtain the NC membrane modified by the nano fibers.
Example 3
The immunochromatographic membrane is obtained by an electrostatic spinning method with simple operation, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 25%;
2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5: 3;
step 3, preparing a spinning solution: respectively putting cellulose nitrate, a high-volatility polar organic solvent and a high-conductivity polar solvent into a small beaker, and stirring for 24 hours at room temperature to obtain a light yellow transparent clear solution to be used for spinning;
step 4, electrostatic spinning operation: the solution was sonicated in a sonicator for 0.5h until the bubbles in the liquid were completely removed. Carefully placing a commercially available NC membrane on a flat plate receiver, then injecting a spinning solution into a 10mL syringe, installing a needle and a pipe, finally installing the needle and the pipe on an electrostatic spinning machine, starting an instrument to spin, uniformly distributing and covering spinning nano fibers on the surface of the NC membrane, and spinning for 30 min; the spinning voltage is 18 KV; the injection speed of the spinning solution is 1.5 mL/h; the diameter of the spray head is 0.8 mm; the moving speed is 30mm/s, and the reciprocating distance is 220 mm;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ for 8h to obtain the NC film modified by the nano fibers.
Example 4
The immunochromatographic membrane is obtained by an electrostatic spinning method with simple operation, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 28%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5: 2;
step 3, preparing a spinning solution: respectively putting cellulose nitrate, a high-volatility polar organic solvent and a high-conductivity polar solvent into a small beaker, and stirring for 24 hours at room temperature to obtain a light yellow transparent clear solution to be used for spinning;
step 4, electrostatic spinning operation: the solution was sonicated in a sonicator for 0.5h until the bubbles in the liquid were completely removed. Carefully placing a commercially available NC membrane on a flat plate receiver, then injecting a spinning solution into a 10mL syringe, installing a needle and a pipe, finally installing the needle and the pipe on an electrostatic spinning machine, starting an instrument to spin, uniformly distributing and covering spinning nano fibers on the surface of the NC membrane, and spinning for 40 min; the spinning voltage is 22 KV; the injection speed of the spinning solution is 2.0 mL/h; the diameter of the spray head is 0.9 mm; the moving speed is 40mm/s, and the reciprocating distance is 240 mm;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ for 10 hours to obtain the NC film modified by the nano fibers.
Example 5
The immunochromatographic membrane is obtained by an electrostatic spinning method with simple operation, and is specifically implemented according to the following steps:
step 1, accurately weighing nitrocellulose, wherein the mass percent of the nitrocellulose is 32%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5: 1;
step 3, preparing a spinning solution: respectively putting cellulose nitrate, a high-volatility polar organic solvent, a high-conductivity polar solvent and a non-ionic surfactant into a small beaker, and stirring at room temperature for 24 hours to obtain a light yellow transparent clear solution to be used for spinning;
step 4, electrostatic spinning operation: the solution was sonicated in a sonicator for 2.5h until the bubbles in the liquid were completely removed. Carefully placing a commercially available NC membrane on a flat plate receiver, then injecting a spinning solution into a 10mL syringe, installing a needle and a pipe, finally installing the needle and the pipe on an electrostatic spinning machine, starting an instrument to spin, uniformly distributing and covering spinning nano fibers on the surface of the NC membrane, and spinning for 1 h; the spinning voltage is 30 KV; the injection speed of the spinning solution is 3.0 mL/h; the diameter of the spray head is 0.1 mm; the moving speed is 40mm/s, and the reciprocating distance is 240 mm;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ for 15 hours to obtain the NC film modified by the nano fibers, wherein the performances of the NC film are shown in Table 1.
TABLE 1 Properties of NC fiber coating films prepared by electrospinning
| Performance of | Flow rate/s | Porosity/% |
| Commercially available NC | 58 | 39 |
| Example 1 | 73 | 54 |
| Example 2 | 76 | 59 |
| Example 3 | 78 | 63 |
| Example 4 | 77 | 62 |
| Example 5 | 75 | 64 |
The invention relates to a preparation method of a nanofiber modified cellulose nitrate immunochromatographic membrane, which comprises the following steps of firstly, accurately weighing cellulose nitrate, accurately weighing N, N-dimethylformamide and acetone, respectively adding the N, N-dimethylformamide and acetone into a beaker, and uniformly stirring to obtain a spinning solution; commercially available NC membranes were then carefully placed on the plate receiver; then starting an instrument to spin and uniformly distributing and covering the spinning nano fibers on the surface of the NC membrane; and after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ to obtain the NC film modified by the nano fibers. Compared with a commercially available NC membrane, the membrane prepared by the invention has the advantages that the slow flow rate and the increased porosity of the solution on the surface of the membrane can be visually seen from figure 1, figure 2 and table 1; and as can be seen from the comparison between fig. 3 and fig. 4, the immunochromatographic membrane of the present invention is a hydrophobic membrane, which is beneficial to reduce the flow rate of the target object to be detected on the membrane surface and increase the interaction rate of the target object to be detected and the detection probe, and these results all indicate that the performance of the present invention is excellent. The immunochromatographic test strip has the characteristics of low flow rate and high porosity, is favorable for increasing the time of antigen-antibody specific reaction, achieves the aim of improving the sensitivity of the test strip, and has good social benefit in instant detection. The nanofiber modified cellulose nitrate immunochromatographic membrane is prepared by an electrostatic spinning method, and the porosity and the flow rate of the surface of the membrane are adjusted by regulating and controlling the electrostatic spinning time and parameters, so that the protein adsorption performance is improved, and the opportunity of antigen-antibody reaction is increased. Experiments prove that the immunochromatographic membrane has higher sensitivity, specificity and repeatability when used as an immunochromatographic test strip, and can be used for preparing a high-sensitivity immunochromatographic membrane.
Claims (6)
1. A preparation method of a nanofiber-modified cellulose nitrate immunochromatographic membrane is characterized by comprising the following steps:
step 1, accurately weighing nitrocellulose according to the total required amount of a spinning solution, wherein the mass percent of nitrocellulose serving as a solute in the spinning solution is 15-40%;
step 2, accurately measuring a high-volatility polar organic solvent and a high-conductivity polar solvent, wherein the volume ratio of the organic solvent to the polar solvent is 5:1-1: 5;
step 3, preparing a spinning solution: adding cellulose nitrate, an organic solvent and a polar solvent into a container, and uniformly stirring to obtain a light yellow transparent clear spinning solution;
step 4, electrostatic spinning operation: a commercially available Nitrocellulose (NC) membrane was carefully placed on a plate receiver, and the spinning solution was sonicated to remove air bubbles from the solution; then injecting the solution into an injector and then installing the injector on an electrostatic spinning machine, setting spinning voltage, spinning solution injection speed and the angle between a needle head and a flat plate, starting an instrument for spinning, and enabling the spinning nanofibers to be uniformly distributed and covered on the surface of the NC membrane;
and 5, after spinning is finished, drying the NC film with the surface covered with the nano fibers in a vacuum drying oven at 40 ℃ to obtain the NC film modified by the nano fibers.
2. The method for preparing a nanofiber-modified nitrocellulose immunochromatographic membrane according to claim 1, wherein in the step 2, the organic solvent is acetone.
3. The method for preparing a nanofiber-modified nitrocellulose immunochromatographic membrane according to claim 1, wherein in the step 2, the polar solvent is N, N-dimethylformamide.
4. The method for preparing the nanofiber-modified cellulose nitrate immunochromatographic membrane according to claim 1, wherein in the step 4, the electrospinning time is 0-4 h.
5. The preparation method of the nanofiber-modified nitrocellulose immunochromatographic membrane according to claim 1, wherein in the step 4, the spinning voltage is 10-30 KV; the injection speed of the spinning solution is 0.1-3 mL/h; the diameter of the spray head is 0.1-1 mm; the moving speed is 10-40mm/s, and the reciprocating distance is 180-240 mm.
6. The method for preparing the nanofiber-modified cellulose nitrate immunochromatographic membrane according to claim 1, wherein in the step 5, the drying temperature is not more than 40 ℃ and the drying time is varied from 4 to 24 hours according to the thickness of the spinning membrane.
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