CN105928773B - A method of the quickly and efficiently concentration charge-carrying component on paper base analytical equipment - Google Patents
A method of the quickly and efficiently concentration charge-carrying component on paper base analytical equipment Download PDFInfo
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- CN105928773B CN105928773B CN201610257918.3A CN201610257918A CN105928773B CN 105928773 B CN105928773 B CN 105928773B CN 201610257918 A CN201610257918 A CN 201610257918A CN 105928773 B CN105928773 B CN 105928773B
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- 238000000034 method Methods 0.000 title claims abstract description 38
- 239000000463 material Substances 0.000 claims abstract description 35
- 239000012488 sample solution Substances 0.000 claims abstract description 33
- 239000003792 electrolyte Substances 0.000 claims abstract description 29
- 238000012360 testing method Methods 0.000 claims abstract description 20
- 239000012530 fluid Substances 0.000 claims abstract description 6
- 239000000523 sample Substances 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 22
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims description 19
- 230000005684 electric field Effects 0.000 claims description 14
- 230000008569 process Effects 0.000 claims description 11
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 7
- 238000005370 electroosmosis Methods 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 239000008151 electrolyte solution Substances 0.000 claims description 5
- 239000000835 fiber Substances 0.000 claims description 5
- 239000003365 glass fiber Substances 0.000 claims description 4
- 230000002209 hydrophobic effect Effects 0.000 claims description 4
- 229940020947 fluorescein sodium Drugs 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 229920002301 cellulose acetate Polymers 0.000 claims description 2
- 229920005594 polymer fiber Polymers 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- -1 concentration 200mM Substances 0.000 claims 1
- 239000004753 textile Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 10
- 238000001514 detection method Methods 0.000 abstract description 8
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000012545 processing Methods 0.000 abstract description 2
- 229960002143 fluorescein Drugs 0.000 description 16
- 230000033001 locomotion Effects 0.000 description 11
- 230000009471 action Effects 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 239000004744 fabric Substances 0.000 description 6
- 229920000742 Cotton Polymers 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 235000009328 Amaranthus caudatus Nutrition 0.000 description 3
- 240000001592 Amaranthus caudatus Species 0.000 description 3
- 235000012735 amaranth Nutrition 0.000 description 3
- 239000004178 amaranth Substances 0.000 description 3
- 239000000428 dust Substances 0.000 description 3
- 230000005611 electricity Effects 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000003321 amplification Effects 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 238000002218 isotachophoresis Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 1
- CGNLCCVKSWNSDG-UHFFFAOYSA-N SYBR Green I Chemical compound CN(C)CCCN(CCC)C1=CC(C=C2N(C3=CC=CC=C3S2)C)=C2C=CC=CC2=[N+]1C1=CC=CC=C1 CGNLCCVKSWNSDG-UHFFFAOYSA-N 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
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- 238000004737 colorimetric analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
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- 238000002474 experimental method Methods 0.000 description 1
- 239000002657 fibrous material Substances 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000002032 lab-on-a-chip Methods 0.000 description 1
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- 238000012544 monitoring process Methods 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 238000004094 preconcentration Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Classifications
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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
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of quickly and efficiently method of concentration charge-carrying component on paper base analytical equipment, circuit is formed using paper base material, background electrolyte, testing sample solution, two electrodes and DC power supply, two of them electrode is inserted into background electrolyte and testing sample solution respectively, and the both ends of paper base material are contacted with background electrolyte and testing sample solution respectively.The present invention has many advantages, such as that easy to operate, concentration is high-efficient, quick and low in cost, can effectively improve detection sensitivity for the determinand sample of low concentration, and without processing complicated fluid channel.
Description
Technical field
The invention belongs to technical field of analytical chemistry, and in particular to a kind of quickly and efficiently concentration on paper base analytical equipment
The method of charge-carrying component.
Background technique
It is the hot spot of nearest analysis science with the paper base analytical equipment that micro-fluidic paper chip represents, this kind of analytical equipment tools
Have the advantages that raw material sources are wide, cheap, degradable, there are capillarity, portable and biochemistry compatibility are good.Pass through
The fluid channel of certain structure is processed on paper, solution can be by the capillarity directed flow on paper, and then realizes analysis
The automation and micromation of detection.With going deep into for research, the diversification of detection means, application range is extended from medical analysis
To numerous areas such as environmental monitoring, food safety, cell analysis, immunoassay, bio-sensings, detected by analysis and bed at the scene
In great application potential.
However limiting the principal element that paper base analytical equipment further develops is its lower sensitivity, it is especially right
Some low-abundance target analytes, this kind of analytical equipment cannot provide corresponding testing result.This greatly affected paper base
The application range and potentiality of analytical equipment.
In order to improve the sensitivity of paper base analytical equipment, the amplification mechanism of some chemistry and biology[1-2]It is introduced in paper base
In analytical equipment, but the introducing of these mechanism is along with the materials synthesis step for using expensive biochemical reagents and complicated and time consumption
Suddenly, the use cost and time cost of paper base analytical equipment are improved.The method that another kind improves the sensitivity of paper base analytical equipment
It is to be realized by the method for sample preconcentration, including evaporate solvent and electronic concentration.Evaporation solvent need external heat source and
It is not suitable for thermally sensitive analyte；Electronic concentration is to improve a kind of method of capillary electrophoresis detection sensitivity, is had
Operation mode multiplicity, the characteristics of having a wide range of application, principle is to charge target analytes movement velocity in tubing string by changing
Variation, cause pile up effect, so achieve the purpose that improve sensitivity for analysis.Report at present on paper base analytical equipment
Electronic concentration mode only has isotachophoresis[3]With ion concentration polarity effect[4], isotachophoresis procedures' needs are for band electroanalysis
The mobility selection of object is suitably leading to lead electrolyte with after, and operation does not have flexibility；And based on ion concentration polarity effect
Concentration process needs introduce ion selective membrane on fluid channel, complicated for operation, and long the time required to concentration.Therefore it needs
Efficient one kind, quick, low cost, the simply electronic concentration process of operation mode is developed to analyze with lower at original raising paper base
The sensitivity of device, to enhance its application potential.
Summary of the invention
The quickly and efficiently method of concentration charge-carrying component that the purpose of the present invention is to provide a kind of on paper base analytical equipment,
Keep change of the target analytes of electrification under the convection action of electric field, capillarity and electroosmotic flow due to movement velocity fast
The concentration of fast ground improves the detection sensitivity of paper base analytical equipment in a certain position.
To achieve the goals above, the technical solution adopted by the present invention is that:
A method of the quickly and efficiently concentration charge-carrying component on paper base analytical equipment, using paper base material, background electricity
Electrolyte solution, testing sample solution, two electrodes and DC power supply form circuit, and two of them electrode is inserted into background electricity respectively
In electrolyte solution and testing sample solution, the both ends of paper base material connect with background electrolyte and testing sample solution respectively
Touching.
In the above method, described two electrodes are connect by conducting wire with DC power supply.
In the above method, the length of the paper base material is 1 ~ 10cm, and 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 fabric sheet
Material or polymer fiber.
In the above method, for the fluid channel of background electrolyte and testing sample solution flowing on the paper base material
It can be formed by directly cutting out paper base material, either be applied on paper base material using hydrophobic material such as hydrophobic film or wax
It is conformal at.
In the above method, the ratio between conductivity of the background electrolyte and testing sample solution is greater than 1.
In the above method, the electric field strength being applied on paper base material is 10 ~ 500V/cm.
In the above method, electroosmotic flow regulator can also be added in background electrolyte and testing sample solution,
To change electroosmotic flow size and direction on paper base material.
The principle of the present invention is as follows: after background electrolyte and the contact of testing sample solution and paper base material, in capillary
It is moved toward one another on paper base material under effect, when applying DC voltages to two kinds of solution, and two solution are on paper base material
Circuit is connected when contact, and the electrification target analytes in testing sample solution are due to sample solution and background electrolyte
The difference of region field distribution and the convection action of electroosmotic flow cause movement velocity to change, to generate the heap of target analytes
Product effect, makes target analytes concentration on paper base material.
The invention has the benefit that
The present invention has many advantages, such as that easy to operate, concentration is high-efficient, quick and low in cost, for low concentration to
Surveying object sample can effectively improve detection sensitivity, and without processing complicated fluid channel.By taking fluorescein molecule as an example, In
Under the low consistency conditions of 1 ~ 10nmol/L, sensitivity can be improved using the present invention by least three order of magnitude or more.In addition, this
Invention can also be combined with other detection methods, such as colorimetric method, Electrochemical Detection, fluorescence detection, mass spectrum and chemiluminescence
Deng for developing sensitive quick paper base analytical equipment with very big application value.
Detailed description of the invention
Fig. 1 is the structural schematic diagram that the present invention implements the paper base analytical equipment used.
Fig. 2 is believe in electronic concentrating process to fluorescein molecule using the electronic concentration process of the embodiment of the present invention 1
Number intensity and concentration multiple change with time relational graph.
Fig. 3 is the fluorescence photo of fluorescein molecule concentration band in the embodiment of the present invention 1.
Fig. 4 is to carry out signal in electronic concentrating process to λ DNA molecular using the electronic concentration process of the embodiment of the present invention 2
Intensity changes with time relational graph.
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.
Specific embodiment
The present invention is described in further details below with reference to specific embodiment, it is described be explanation of the invention without
It is to limit.
Embodiment 1
The quickly and efficiently method of concentration fluorescein molecule on paper base analytical equipment as shown in Figure 1, using 2mM's
Tris-HCl solution prepares the fluorescein sodium sample solution that initial concentration is 5nM, is inserted into electrode as cathode, with 200 mM
Tris-HCl solution is inserted into electrode as anode, two electrodes are connect by conducting wire with DC power supply as background electrolyte；
Cellulosic filter paper paraffin is applied into growth 3cm, the strip of wide 2mm and along its length both ends open opens cellulosic filter paper
The both ends put are contacted with background electrolyte and testing sample solution respectively, and two kinds of solution are made to transport in opposite directions under capillary forces
It is dynamic, power on, apply the DC voltage of 300V, fluorescein molecule under the action of electric field to anode movement, since sample is molten
The electric field strength in liquid zone domain be higher than background electrolyte region electric field strength, when fluorescein molecule by sample solution region into
Movement velocity reduction causes to accumulate when entering into background electrolyte, forms concentration band to the greatest extent.Using inverted fluorescence microscope
Entire concentrating process is observed, and fluorescence imaging is carried out to concentration region every 20 s, the photo of acquisition uses ImageJ software
Processing.
Experimental result is shown: Fig. 2 be the collected fluorescein concentration band signal intensity of the present embodiment and concentration multiple at any time
Between the curve relation figure that changes；Fig. 3 is the fluorescence photo of fluorescein molecule concentration band, can be seen that by Fig. 2 and Fig. 3 and works as concentration
When time reaches 220 s, concentration multiple just has been over 1000 times, and concentration multiple is stablized between 220 ~ 300 s
1000 times or so.Experimental result illustrates that the method for the present embodiment electronic concentration fluorescein on cellulosic filter paper is efficiently quick.
Embodiment 2
The quickly and efficiently method of concentration DNA molecular on paper base analytical equipment as shown in Figure 1, using the Tris- of 2mM
HCl solution prepares initial concentration and is the λ DNA solution of 4pg/ μ L, and SYBR Green I is added makes its concentration 0.4X, as sample
Product solution is inserted into electrode as cathode, is added using 200 mM Tris-HCl solution as background electrolyte, and thereto
2%(mass/volume) polyvinylpyrrolidone as electroosmotic flow regulator, to reduce the electric osmose flow horizontal on paper base material, insert
Enter electrode as anode, two electrodes are connect by conducting wire with DC power supply；Glass fibre membrane is cut into growth 4cm, the item of wide 2mm
Shape, both ends are contacted with background electrolyte and testing sample solution respectively, and two kinds of solution are made to transport in opposite directions under capillary forces
It is dynamic, power on, apply the DC voltage of 300V, DNA molecular under the action of electric field to anode movement, due to sample solution area
The electric field strength in domain is higher than the electric field strength in background electrolyte region, when DNA molecular enters back by sample solution region
Movement velocity reduction causes to accumulate when in scape electrolyte solution, forms concentration band to the greatest extent.Using being inverted, fluorescent fiber sem observation is whole
A concentrating process, and fluorescence imaging is carried out to concentration region every 20 s, the photo of acquisition is handled using ImageJ software.
Experimental result is shown: Fig. 4 is that the collected DNA concentration band signal intensity of the present embodiment and concentration multiple become at any time
The curve relation figure of change；Fig. 5 is the fluorescence photo of DNA molecular concentration band, can be seen that initial concentration is 4 by Fig. 4 and Fig. 5
For the λ DNA of pg/ μ L after concentration, the fluorescence intensity after 140s has been more than 4 ng/ μ L, and concentration multiple is more than 1000 times.
Experimental result illustrates that the method for the present embodiment electronic concentrating nucleic acid molecules on glass fibre membrane is efficiently quick, in biomedicine
With very big application potential.
Embodiment 3:
The quickly and efficiently method of concentration fluorescein molecule on paper base analytical equipment as shown in Figure 1, using 2mM's
The fluorescein sodium sample solution that Tris-HCl solution compound concentration is 1 μM is inserted into electrode as anode, with 200 mM Tris-
HCl solution is inserted into electrode as cathode, two electrodes are connect by conducting wire with DC power supply as background electrolyte；With length
For 3cm, the cotton thread that diameter is 1mm is paper base material, and cotton thread both ends connect with background electrolyte and testing sample solution respectively
Touching, two kinds of solution move toward one another under capillary forces, power on, and apply the DC voltage of 300V, fluorescein molecule exists
To cathode motion under the action of electric field, the electricity with background electrolyte region is higher than due to the electric field strength in sample solution region
Field intensity, when fluorescein molecule is entered in background electrolyte by sample solution region, due to the reduction of movement velocity
It causes to accumulate, forms concentration band to the greatest extent.Fig. 6 is the concentration band photo that camera takes, in 120s, fluorescein molecule on cotton thread
Concentration band it is high-visible.The experimental results showed that the method for the present embodiment electronic concentration fluorescein on cotton thread is efficiently quick.Together
When also illustrate, concentration process of the invention can be applied to using cotton thread as on the analytical equipment of base material, have to fiber base material
There is wider compatibility.
Embodiment 4
The quickly and efficiently method of concentration fluorescein molecule on paper base analytical equipment as shown in Figure 1, using deionization
Water prepare initial concentration be 1 μ g/L edible pigment --- amaranth solution inserts electrodes into sample solution as sample solution
It is middle to be used as anode, using the Tris-HCl solution of 300mM as background electrolyte, and inserts electrodes into and be wherein used as cathode,
With a length of 5cm, polyester fiber non-dust cloth that width is 4mm as paper base material, the both ends of non-dust cloth respectively with sample solution and back
Scape electrolyte solution contacts, two kinds of solution move toward one another under capillary action, power on and apply the DC voltage of 500V, amaranth
The red molecule of dish under the action of electric field to cathode motion, since the electric field strength in sample solution region is higher than background electrolyte
The electric field strength in region, when amaranth molecule is entered in background electrolyte by sample solution region, movement velocity is reduced
It causes to accumulate, forms concentration band to the greatest extent.It can see in 130s and occur clearly red concentration band on non-dust cloth.Experiment knot
Fruit shows that electronic concentration process proposed by the present invention can be applied to using cloth as on the analytical equipment of base material, and this
Embodiment electronic concentration edible pigment method on cloth is efficiently quick, has in edible safety detection field huge using valence
Value.
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 (2)
1. a kind of quickly and efficiently method of concentration charge-carrying component on paper base analytical equipment, it is characterised in that use paper base material
Material, background electrolyte, testing sample solution, two electrodes and DC power supply form circuit, two of them electrode difference
It is inserted into background electrolyte and testing sample solution, the both ends of paper base material are respectively with background electrolyte and to test sample
The contact of product solution；In the above process, by directly cutting out paper base material or coated with hydrophobic material is formed on paper base material
For the fluid channel that background electrolyte and testing sample solution flow, and the electric field strength being applied on paper base material is
75 ~ 100 V/cm, in which: the length of the paper base material is 1 ~ 10 cm, and material is cellulosic filter paper, glass fibre membrane, nitrification
Tunica fibrosa, cellulose acetate film, fiber wire rod, strip nonwoven or textile sheet or polymer fiber；The hydrophobic material is thin
Water film or wax；The ratio between conductivity of the background electrolyte and testing sample solution is greater than 1, and the back-ground electolyte is molten
Liquid be Tris-HCl solution, concentration 200mM, sample solution be Fluress or λ DNA solution, when for fluorescein sodium it is molten
When liquid, concentration is 5 nM, and when for λ DNA solution, concentration is 4pg/ μ L；Through detecting, 140 ~ 220s of concentration, concentration multiple is more than
1000 times.
2. a kind of quickly and efficiently method of concentration charge-carrying component on paper base analytical equipment according to claim 1,
Be characterized in that can adding in the background electrolyte and sample solution to change on paper base material electroosmotic flow size and
The electroosmotic flow regulator in direction.
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