CN105928773A - Method for rapid high-efficiency concentration of charged components on paper-based analytical apparatus - Google Patents
Method for rapid high-efficiency concentration of charged components on paper-based analytical apparatus Download PDFInfo
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- CN105928773A CN105928773A CN201610257918.3A CN201610257918A CN105928773A CN 105928773 A CN105928773 A CN 105928773A CN 201610257918 A CN201610257918 A CN 201610257918A CN 105928773 A CN105928773 A CN 105928773A
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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
- G01N1/40—Concentrating samples
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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
- G01N1/40—Concentrating samples
- G01N2001/4038—Concentrating samples electric methods, e.g. electromigration, electrophoresis, ionisation
Abstract
The invention provides a method for rapid high-efficiency concentration of charged components on a paper-based analytical apparatus. According to the method, a loop is composed of a paper-based material, a background electrolyte solution, a to-be-detected sample solution, two electrodes and a direct-current source, wherein the two electrodes are respectively inserted into the background electrolyte solution and the to-be-detected sample solution, and two ends of the paper-based material respectively contact with the background electrolyte solution and the to-be-detected sample solution. The method has the advantages of simple operation, high concentration efficiency, rapidness, low cost and the like; and directed at a to-be-detected sample with low concentration, the method has improved detection sensitivity and does not need processing of a complex fluid flow passage.
Description
Technical field
The invention belongs to technical field of analytical chemistry, be specifically related to a kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment.
Background technology
The paper substrate analytical equipment represented with micro-fluidic paper chip is the focus of nearest analysis science, and this alanysis device has that raw material sources are wide, cheap, degradable, there is the advantages such as capillarity, portable and biochemistry compatibility be good.By processing the fluid passage of a fixed structure on paper, solution can rely on the capillarity directed flow on paper, and then realizes analyzing automation and the miniaturization of detection.Along with going deep into of research, the variation of detection means, its range of application expands to the numerous areas such as environmental monitoring, food security, cell analysis, immunoassay, bio-sensing from medical analysis, analyzes at the scene and great application potential in the other detection of bed.
But the principal element that restriction paper substrate analytical equipment develops further is its relatively low sensitivity, especially to some low-abundance target analytes, this kind of analytical equipment can not provide corresponding testing result.This range of application that greatly have impact on paper substrate analytical equipment and potentiality.
In order to improve the sensitivity of paper substrate analytical equipment, some chemistry and biological amplification mechanism[1-2]It is introduced in paper substrate analytical equipment, but introducing along with using expensive biochemical reagents and the materials synthesis step of complicated and time consumption of these mechanism, improve use cost and the time cost of paper substrate analytical equipment.The another kind of method improving paper substrate analytical equipment sensitivity is to be realized by the method for sample preconcentration, including evaporation solvent and electronic concentration.Evaporation solvent needs external heat source and is not suitable for thermally sensitive analyte；Electronic concentration is a kind of method improving Capillary Electrophoresis detection sensitivity, there is operator scheme various, the feature of applied range, its principle is by changing the change of movement velocity in tubing string of charged target analytes, cause pile up effect, and then reach to improve the purpose of sensitivity for analysis.The electronic concentration pattern on paper substrate analytical equipment of report only has isotachophoresis at present[3]With ion concentration polarity effect[4], isotachophoresis procedures be required for the mobility of charged analyte select the most leading and after lead electrolyte, operation does not have flexibility；And concentration process of based on ion concentration polarity effect needs to introduce ion selective membrane, operation complexity on fluid passage, and concentration required time is long.Therefore the sensitivity that exploitation is a kind of efficiently, quick, the simple electronic concentration process of low cost, operator scheme originally improves paper substrate analytical equipment with relatively low becoming is needed badly, to strengthen its application potential.
Summary of the invention
It is an object of the invention to provide a kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment, make charged target analytes under the convection action of electric field, capillarity and EOF owing to the change concentration rapidly of movement velocity is in a certain position, and then improve the detection sensitivity of paper substrate analytical equipment.
To achieve these goals, the technical solution used in the present invention is:
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment, use paper base material, background electrolyte, testing sample solution, two electrodes and dc source composition loop, two of which electrode inserts in background electrolyte and testing sample solution respectively, and the two ends of paper base material contact with background electrolyte and testing sample solution respectively.
In said method, said two electrode is connected with dc source by wire.
In said method, a length of 1 ~ the 10cm of described paper base material, material is fibrous material, including cellulosic filter paper, chromatographic paper, office printing paper, glass fibre membrane, nitrocellulose membrane, cellulose acetate film, fiber wire rod, strip nonwoven or textile sheet or polymer fiber.
In said method, on described paper base material, the fluid passage for background electrolyte and testing sample solution flowing by paper base material is directly cut out formation, or can use the coating of hydrophobic material such as hydrophobic film or wax to be formed on paper base material.
In said method, the ratio of the electrical conductivity of described background electrolyte and testing sample solution is more than 1.
In said method, putting on the electric-field intensity on paper base material is 10 ~ 500V/cm.
In said method, it is also possible to add EOF conditioning agent in background electrolyte and testing sample solution, in order to change the EOF size on paper base material and direction.
The principle of the present invention is as follows: after background electrolyte contacts with paper base material with testing sample solution, move toward one another is made under capillary action on paper base material, when applying DC voltage to two kinds of solution, and circuit turn-on when that two solution contacting on paper base material, charged target analytes in testing sample solution causes movement velocity to change due to the convection action of sample solution and the difference of background electrolyte region Electric Field Distribution and EOF, thus produce the pile up effect of target analytes, make target analytes concentration on paper base material.
The invention have the benefit that
The present invention has that simple to operate, concentration efficiency is high, quickly and the advantage such as with low cost, and the determinand sample for low concentration can be effectively improved detection sensitivity, and without the fluid passage of processed complex.As a example by fluorescein molecule, under the low consistency conditions of 1 ~ 10nmol/L, use the present invention sensitivity can be improved more than at least 3 orders of magnitude.It is also possible to combine with other detection method, such as colorimetric method, Electrochemical Detection, fluoroscopic examination, mass spectrum and chemiluminescence etc., it has the biggest using value for developing sensitive quick paper substrate analytical equipment.
Accompanying drawing explanation
Fig. 1 is the structural representation that the present invention implements the paper substrate analytical equipment used.
Fig. 2 is that the electronic concentration process using the embodiment of the present invention 1 carries out signal strength signal intensity and concentration multiple graph of a relation over time in electronic concentrating process to fluorescein molecule.
Fig. 3 is the fluorescence photo of fluorescein molecule concentration band in the embodiment of the present invention 1.
Fig. 4 is that the electronic concentration process using the embodiment of the present invention 2 carries out signal strength signal intensity graph of a relation over time in electronic concentrating process to λ DNA molecular.
The micro-imaging photo of λ DNA molecular concentration band in Fig. 5 embodiment of the present invention 2.
Fig. 6 is the fluorescence photo of fluorescein molecule concentration band in the embodiment of the present invention 3.
Detailed description of the invention
Being described in further details the present invention below in conjunction with specific embodiment, the explanation of the invention is not limited.
Embodiment 1
The method of quickly and efficiently concentration fluorescein molecule on paper substrate analytical equipment as shown in Figure 1, uses the fluorescein sodium sample solution that Tris-HCl solution preparation initial concentration is 5nM of 2mM, inserts electrode as negative electrode, with 200 mM
Tris-HCl solution is as background electrolyte, and insertion electrode is as anode, and two electrodes are connected with dc source by wire；Cellulosic filter paper paraffin is applied growth 3cm, the strip of wide 2mm and along its length both ends open, the two ends that cellulosic filter paper is open are contacted with background electrolyte and testing sample solution respectively, two kinds of solution make move toward one another under capillary forces, switch on power, apply the DC voltage of 300V, fluorescein molecule under the effect of electric field to anode movement, electric-field intensity due to sample solution region is higher than the electric-field intensity in background electrolyte region, when fluorescein molecule is entered in background electrolyte by sample solution region, movement velocity reduction causes accumulation, form concentration band to the greatest extent.Using inverted fluorescence microscope to observe whole concentrating process, and every 20 s, concentration region is carried out fluorescence imaging, the photo of collection uses ImageJ software to process.
Experimental result shows: Fig. 2 is the fluorescein concentration band signal intensity that collects of the present embodiment and the time dependent curve relation figure of concentration multiple；Fig. 3 is the fluorescence photo of fluorescein molecule concentration band, can be seen that, when the concentration time reaches 220 s, concentration multiple just has been over 1000 times by Fig. 2 and Fig. 3, and concentration multiple is stable at about 1000 times between 220 ~ 300 s.Experimental result illustrates that the present embodiment method of electronic concentration fluorescein on cellulosic filter paper is the quickest.
Embodiment 2
The method of quickly and efficiently concentration DNA molecular on paper substrate analytical equipment as shown in Figure 1, use the λ DNA solution that Tris-HCl solution preparation initial concentration is 4pg/ μ L of 2mM, and add SYBR Green I and make its concentration be 0.4X, as sample solution, insertion electrode is as negative electrode, with 200 mM
Tris-HCl solution is as background electrolyte, and be added thereto to 2%(mass/volume) polyvinylpyrrolidone as EOF conditioning agent, to reduce the EOF level on paper base material, insertion electrode is as anode, and two electrodes are connected with dc source by wire；By glass fibre membrane cutting growth 4cm, the strip of wide 2mm, two ends contact with background electrolyte and testing sample solution respectively, two kinds of solution make move toward one another under capillary forces, switch on power, apply the DC voltage of 300V, DNA molecular under the effect of electric field to anode movement, electric-field intensity due to sample solution region is higher than the electric-field intensity in background electrolyte region, when DNA molecular is entered in background electrolyte by sample solution region, movement velocity reduction causes accumulation, forms concentration band to the greatest extent.Using and be inverted the whole concentrating process of fluorescent fiber sem observation, and every 20 s, concentration region is carried out fluorescence imaging, the photo of collection uses ImageJ software to process.
Experimental result shows: Fig. 4 is the DNA concentration band signal intensity that collects of the present embodiment and the time dependent curve relation figure of concentration multiple；Fig. 5 is the fluorescence photo of DNA molecular concentration band, by Fig. 4 and Fig. 5 can be seen that λ DNA that initial concentration is 4 pg/ μ L after concentration, the fluorescence intensity after 140s has exceeded 4 ng/ μ L, and its concentration multiple is more than 1000 times.Experimental result illustrates that the present embodiment method of electronic concentrating nucleic acid molecules on glass fibre membrane is efficiently quick, has the biggest application potential in biomedicine.
Embodiment 3:
The method of quickly and efficiently concentration fluorescein molecule on paper substrate analytical equipment as shown in Figure 1, uses the fluorescein sodium sample solution that Tris-HCl solution compound concentration is 1 μM of 2mM, inserts electrode as anode, with 200 mM
Tris-HCl solution is as background electrolyte, and insertion electrode is as negative electrode, and two electrodes are connected with dc source by wire；With a length of 3cm, the cotton thread of a diameter of 1mm is paper base material, cotton thread two ends contact with background electrolyte and testing sample solution respectively, two kinds of solution make move toward one another under capillary forces, switch on power, apply the DC voltage of 300V, fluorescein molecule under the effect of electric field to cathode motion, owing to the electric-field intensity in sample solution region is higher than the electric-field intensity with background electrolyte region, when fluorescein molecule is entered in background electrolyte by sample solution region, owing to the reduction of movement velocity causes accumulation, form concentration band to the greatest extent.Fig. 6 is the concentration band photo that camera photographs, and when 120s, on cotton thread, the concentration band of fluorescein molecule is high-visible.Test result indicate that the present embodiment method of electronic concentration fluorescein on cotton thread is the quickest.The most also illustrating, the concentration process of the present invention may apply on the analytical equipment with cotton thread as base material, has wider compatibility to fiber base material.
Embodiment 4
nullThe method of quickly and efficiently concentration fluorescein molecule on paper substrate analytical equipment as shown in Figure 1，Using deionized water preparation initial concentration is that the food coloring amaranth solution of 1 μ g/L is as sample solution，Insert electrodes in sample solution as anode，Using the Tris-HCl solution of 300mM as background electrolyte，And insert electrodes into wherein as negative electrode，With a length of 5cm，The polyester fiber non-dust cloth of a width of 4mm is as paper base material，The two ends of non-dust cloth contact with sample solution and background electrolyte respectively，Two kinds of solution move toward one another under capillary action，Switch on power and apply the DC voltage of 500V，Amaranth molecule under the effect of electric field to cathode motion，Electric-field intensity due to sample solution region is higher than the electric-field intensity in background electrolyte region，When amaranth molecule is entered in background electrolyte by sample solution region, movement velocity reduction causes accumulation，Form concentration band to the greatest extent.When 130s it can be seen that occur in that red concentration band clearly on non-dust cloth.Test result indicate that, the electronic concentration process that the present invention proposes may apply on the analytical equipment with cloth as base material, and the present embodiment electronic concentration food coloring method on cloth is efficiently quick, has huge using value at edible safety detection field.
Bibliography
[1] Badu-Tawiah A K, Lathwal S, Kaastrup K, et al. Polymerization-based signal
amplification for paper-based immunoassays[J]. Lab on a Chip, 2015, 15(3):
655-659.
[2] Ma C, Li W, Kong Q,
et al. 3D origami electrochemical immunodevice for
sensitive point-of-care testing based on dual-signal amplification strategy[J]. Biosensors and Bioelectronics, 2015, 63: 7-13.
[3]
Moghadam B Y, Connelly K T, Posner J D. Isotachophoretic
preconcenetration on paper-based microfluidic
devices[J]. Analytical chemistry, 2014, 86(12):
5829-5837. [4] Yang R J, Pu H H, Wang H L. Ion
concentration polarization on paper-based microfluidic
devices and its application to preconcentrate dilute
sample solutions[J]. Biomicrofluidics,
2015, 9(1): 014122.
Claims (7)
1. the method for a quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment, it is characterized in that using paper base material, background electrolyte, testing sample solution, two electrodes and dc source composition loop, two of which electrode inserts in background electrolyte and testing sample solution respectively, and the two ends of paper base material contact with background electrolyte and testing sample solution respectively.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 1, it is characterized in that a length of 1 ~ 10 cm of described paper base material, material is fibrous material, including cellulosic filter paper, chromatographic paper, office printing paper, glass fibre membrane, nitrocellulose membrane, cellulose acetate film, fiber wire rod, strip nonwoven or textile sheet or polymer fiber.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 2, it is characterised in that by paper base material directly being cut out or coated with hydrophobic material is formed for background electrolyte and the fluid passage of testing sample solution flowing on paper base material.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 3, it is characterised in that described hydrophobic material is hydrophobic film or wax.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 1, it is characterised in that the ratio of the electrical conductivity of described background electrolyte and testing sample solution is more than 1.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 5, it is characterised in that can add to change the EOF conditioning agent in EOF size and direction on paper base material in described background electrolyte and sample solution.
A kind of method of quickly and efficiently concentration charge-carrying component on paper substrate analytical equipment the most according to claim 1, it is characterised in that putting on the electric-field intensity on paper base material is 10 ~ 500 V/cm.
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| CN115193493A (en) * | 2022-06-10 | 2022-10-18 | 中国科学技术大学 | Paper-based micro-fluidic chip and application thereof |
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| CN115193493B (en) * | 2022-06-10 | 2024-02-09 | 中国科学技术大学 | Paper-based microfluidic chip and application thereof |
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