CN1818662A - Sequential injection analyzer based on capillaries and utilization thereof - Google Patents

Sequential injection analyzer based on capillaries and utilization thereof Download PDF

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Publication number
CN1818662A
CN1818662A CN 200610049830 CN200610049830A CN1818662A CN 1818662 A CN1818662 A CN 1818662A CN 200610049830 CN200610049830 CN 200610049830 CN 200610049830 A CN200610049830 A CN 200610049830A CN 1818662 A CN1818662 A CN 1818662A
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CN
China
Prior art keywords
liquid
coupon
kapillary
capillary
capillaceous
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CN 200610049830
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Chinese (zh)
Inventor
方群
杜文斌
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浙江大学
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Priority to CN 200610049830 priority Critical patent/CN1818662A/en
Publication of CN1818662A publication Critical patent/CN1818662A/en

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Abstract

An analysis device of sequential injection based on capillary tube is composed of three units of capillary tube and sample plate and driving component of liquid flow at capillary tube outlet .The detection, reaction, on-line mixing and leading-in of multisample and multireagent liquid can be realized in capillary tube by utilizing movement of sample tube array.

Description

A kind of based on sequential injection analyzer capillaceous and using method thereof

Technical field

The field that the present invention relates to is a flow analysis, particularly relates to a kind of based on sequential injection device capillaceous and using method thereof.

Background technology

Flow analysis is a kind of automated analysis technology that has grown up since six the seventies, development along with society, traditional is the demand that the pattern hand-manipulated of main tool can't satisfy people gradually with glassware and measuring device, pipeline Continuous Flow Analysis technology occurred.Ruzicka and Hansen have proposed Flow Injection Analysis (Flow injection analysis in 1975, FIA) notion, they have utilized the controllability and the reappearance of liquid level stream mode in the tubule road (<1mm internal diameter), add correct time (being flow velocity) control, realized reproduction but non-admixture completely, and realizing on this basis reappearing and chemical reaction completely. the proposition of this idea has improved analysis speed greatly, make and per hour measure samples up to a hundred and become possibility, also promoted the microminiaturization of analytic system simultaneously.It is that nineteen ninety Ruzicka and Marshall propose that sequential injection is analyzed (Sequential Injection Analysis is called for short SIA).The core component of sequential injection analytic system is a hyperchannel selector valve.Each channel position of this valve links to each other with passages such as detecting device, sample, reagent respectively, and public passage communicates with a pump that can aspirate and promote liquid.By the effect of pump, order is from the district that different passage sucks certain volume takes memotron between pump and the valve to.Then these solution district bands are pushed into detecting device, in pipeline,, radial and axial peptizaiton causes that reagent is with the overlapping of sample band with mix between district's band of sample and reagent in this course owing to interpenetrating, reagent takes place to cause the formation of reaction product than learning reaction with sample.In detecting device, can obtain injecting similar peak type signal in the analysis with proper flow.

20th century, the microfluidic analysis technology based on micro-machined channel network chip appearred in the nineties, the collection of sample, pre-service, analyzing and testing etc. are integrated in several square centimeters the area, can finish sample analysis and detection efficiently, apace, be one of the focus in present analytical chemistry field.Used for reference Flow Injection Analysis at the beginning of the microfluidic analysis development; Through ten years development, microfluidic analysis has had wide development, some flow injections based on microchip (Andrew M.Leach has been proposed simultaneously, Aaron R.Wheeler, and Richard N.Zare.Flow Injection Analysis in a MicrofluidicFormat.Analytical Chemistry, 2003,75:967-972) with (the Richard Davidsson of sequential injection system, Fr é d é ric Genin, Martin Bengtsson, et.al.Microfluidic biosensing systems Part I.Development and optimisation of enzymatic chemiluminescent m-biosensors based onsilicon microchips.Lab On a Chip, 2004,4:481-487).(high flux of microanalysis chip varies device and using method thereof to patent in front continuously, Chinese patent application numbers 200410016224.8) in, proposed a kind of high flux and varied device and using method thereof continuously based on the kapillary sample probe, by processing sampling breach on the sample liquid bath, make that continuous micro-controlled volume sample introduction becomes possibility fast.Based on this technology, set up (the Wen-Bin Du of high flux Flow Injection Analysis system based on micro-fluidic chip, Qun Fang, Qiao-Hong He, Zhao-Lun Fang.High-Throughput Nanoliter Sample Introduction Microfluidic Chip-Based FlowInjection Analysis System with Gravity-Driven Flows.Analytical Chemistry, 2005.77:1330-1337).Be characterized in and rely on gravity and surface tension effects by pump valve, realize automatically Flow Injection Analysis fast; And can realize varying fast continuously of various product.

The system of general flow analysis often adopts mechanical valve system (as sampling valve and multiport valve etc.) to realize operations such as the switching of introducing, reagent, current-carrying of sample and mixing.And the present invention has utilized capillary outlet as the liquid driven end, utilizes the order valveless leading-in end of capillary inlet as sample, reagent and current-carrying.Realized the order introducing of the many liquid of valveless.Based on these characteristics, the structure of system is simplified greatly.And identical microfluidic procedures is if realize needs processing hybrid channel and a plurality of sample reagent liquid bath, and cost height on micro-fluidic chip.

Summary of the invention

The objective of the invention is to break through in the past, sequential injection, flow injection limit to based on the microminiaturization that the design feature of syringe pump and multiport valve and coil pipe is brought, set up a kind of quick sequential injection analytic system based on capillary channel, do not need to adopt expensive complicated multidigit selector valve, the order of the many liquid of upgrading valveless of can realizing receiving is introduced, and finishes the micro-fluidic sequential injection analysis operation of high flux.

The invention provides a kind of based on sequential injection analyzer capillaceous, by kapillary, coupon array liquid drawing-in system, the detection system of sample composition in liquid driven system and the kapillary in the kapillary, it is characterized in that capillary outlet is as the liquid driven end, the capillary inlet is as sample, the order valveless leading-in end of reagent and current-carrying, described capillary inlet end is connected with the coupon outlet of coupon array liquid drawing-in system, coupon array liquid drawing-in system is made up of array and coupon platform that plural coupon constitutes, coupon is fixed on the coupon platform, and capillary outlet is connected with the liquid driven system.

According to the present invention, described kapillary is the main part of system, and liquid flows to endpiece from inlet end, and mixes in kapillary, reacts.The kapillary material can be diversified, comprises quartz, perhaps glass, perhaps metal, perhaps macromolecule polymer material.The capillary channel internal diameter is between 10 nanometers to 5 millimeter, and preferable internal diameter is 5~250 microns.The microcapillary tube wall thickness is in 1 micron to 5 millimeters scope.Capillary pipe length is in 1 millimeter to 10 meters scope, and preferable pipe range scope is 5 millimeters~50 centimetres.

According to the present invention, described coupon is horizontal positioned and is fixed on the coupon platform.Need width of special processing 10 microns to 30 millimeters scopes in the bottom of coupon, the degree of depth exports at 1 micron coupon of coming in and going out for the kapillary entrance point to 5 millimeters scopes.The kind of liquid of packing in the coupon comprises test liquid, reagent solution, carrier fluid.The effect of reagent solution is and sample generation chemical reaction, finishes qualitative and quantitative analysis to sample indirectly by the product of measuring chemical reaction.The kind of reagent solution can have multiple.The effect of carrier fluid is to carry test liquid and reagent solution flows to the capillary outlet end by the kapillary entrance point.Concrete select what class I liquid I, and the putting in order of coupon that these liquid are housed, need decide according to concrete analysis system.Because the yardstick of coupon outlet is less, when the coupon horizontal positioned, relies on capillary effect, the liquid that can not in the coupon of packing into is overflowed by the coupon outlet.The external diameter scope of coupon is 0.5 millimeter to 10 centimetres, and comparatively commonly used is between 2 millimeters to 2 centimetres.The volume range of reprinting liquid in the coupon is 1 to receive and rise to 100 milliliters, has also determined the length of coupon thus.

According to the present invention, in the described coupon array liquid drawing-in system, the coupon platform that coupon is installed can carry out one dimension, perhaps two-dimentional, perhaps San Wei translation motion perhaps can be carried out two dimension or three-dimensional rotation, the compound motion that perhaps can carry out above-mentioned translation and rotation.The purpose that moves is that each coupon on the platform is contacted with the kapillary entrance point in certain sequence, realizes introducing in proper order by pre-programmed many liquid.

According to the present invention, the liquid driven system that adopts in the device is connected with capillary outlet.The direction that drives liquid is to flow or suction to capillary outlet from the kapillary import.Drive system adopts segregation drive power, and perhaps electric osmose drives power, perhaps Mechanical Driven power.Adopt the drive system of Mechanical Driven power to comprise syringe pump, perhaps peristaltic pump, perhaps reciprocating pump.

The usual method of segregation drive stream is that the conduit of capillary outlet end by certain-length linked to each other with a horizontal sewer pipe, and is poor at horizontal sewer pipe and capillary inlet end formation certain liquid level, can realize segregation drive.

The driving flow rates of drive system is 1 a skin liter/minute to 10 ml/min.

According to the present invention, described based on sequential injection analyzer capillaceous in, the method of introducing liquid in kapillary is, mobile coupon platform drives coupon and capillary inlet end in contact, the capillary inlet end immerses certain hour in the sample liquid in pipe by the coupon outlet, under drive systems, liquid flow in the kapillary in the coupon of certain volume.

Adopt said method to carry out liquid and introduce, the volume of introducing liquid in the kapillary is determined the residence time and liquid the flow velocity in kapillary of capillary exit in the sample liquid in pipe by two factors.By changing above-mentioned two factors, can change the volume of introducing liquid.In liquid introducing process, the residence time scope of kapillary import in coupon liquid is 1 millisecond to 60 minutes; The volume range that is introduced into each liquid capillaceous is 1 to ascend to heaven to 500 microlitres.When the kapillary entrance point switches, when flow velocity is very fast, bubble might be sucked influence mensuration in the kapillary between two coupons.The method that solves is to adopt fast switching method, makes at the liquid of kapillary entrance point as yet not in the retraction kapillary and just switch to next coupon before producing bubble; Perhaps utilize the surface tension of liquid at the capillary outlet end, make in handoff procedure, the capillary liquid in pipe keeps arrheaing state.

The using method of sequential injection analyzer is on the kapillary of the present invention, mobile coupon platform drive on it a plurality of coupons in certain sequence successively with the capillary inlet end in contact, the capillary inlet end immerses in the sample liquid in pipe by the coupon import and stops certain hour, under drive systems, make the interior liquid of various sample pipe of certain volume, kind comprises test liquid and carrier fluid at least, be introduced in the kapillary in certain sequence, liquid flows to the capillary outlet end from the capillary inlet end with certain flow rate, in process of flowing, order is introduced between the interior test liquid of kapillary and other liquid physical mixed is taken place, and chemical reaction takes place between the reagent solution, near the capillary outlet end, by to sample itself, perhaps the product of the chemical reaction of sample and reagent detects.

Analysis system to the mensuration of sample can directly be carried out only needs to introduce test liquid and carrier fluid two class I liquid Is (seeing embodiment two) in kapillary in the sequential injection analytic process.For the restriction because of condition, the analysis system that can not directly carry out the mensuration of sample needs the chemical reaction by sample and reagent, finishes qualitative and quantitative analysis to sample indirectly by the product of mensuration chemical reaction.Therefore need in kapillary, to introduce test liquid, carrier fluid and reagent solution three class I liquid Is (seeing embodiment one) in the sequential injection analytic process.

According to the present invention, the detection method for the reaction product of sample in the kapillary adopts detecting pattern on the post usually, perhaps flows out detecting pattern.The former check point is positioned on the kapillary, and the latter detects the sample that is flowed out by capillary outlet, and operable detection technique comprises chemoluminescence method, fluorescence method, photometry, electrochemical method, mass spectroscopy etc.

According to different analysis systems, the kind difference of loading liquid in the coupon, put in order the also difference of while coupon on platform, promptly the order in each liquid introducing kapillary is also different with volume.By a plurality of coupons putting in order on the coupon platform that variety classes liquid is equipped with in change, can change the introducing order of capillary liquid in pipe.The introducing order of different liquid has considerable influence to analytical performance, need select by optimization experiment on concrete experimental system.

Compared with prior art, advantage of the present invention is:

1. the consumption of sample reagent is extremely low, reaches the level of micro-fluidic chip, is suitable for the analysis of the biological sample of the small preciousness of volume.

2. systematic analysis flux height, the favorable reproducibility of analysis.

3. native system has utilized capillary outlet as the liquid driven end, utilizes the order valveless leading-in end of capillary inlet as sample, reagent and current-carrying.Realized the order introducing of the many liquid of valveless.Based on these characteristics, the structure of system is simplified greatly, and system's processing is simple and convenient, and cost is low.

Description of drawings

Fig. 1: structural representation of the present invention;

Fig. 2: the embodiment of the invention one system analyzes the signal record figure that obtains;

Fig. 3: the structural representation of inventive embodiment two;

Fig. 4: adopt preferred embodiment of the present invention two system to analyze the signal record figure that obtains.

Among the figure:

Kapillary 1, capillary inlet end 2, capillary outlet end 3, coupon 4, coupon breach 5, coupon platform 6, standard solution 7, reagent solution 8,9, carrier fluid 10, luminescent detection system 11, luminous mirror 12, plastic flexible pipe 13, sewer pipe 14, deionized water 15, coupon plane matrix rotating disk 16, fluorescein standard solution 17,18,19,20,21, fluorescence detecting system 22,

Embodiment:

Embodiment one

Fig. 1 is one a structural representation according to a preferred embodiment of the invention.Adopt quartz capillary (1) (75 microns of internal diameters; 375 microns of external diameters) constructing system; 5 centimetres of long capillary tubes; the protective layer used sharp pocket knife of kapillary entrance point (2) scrapes off, and kapillary entrance point (2) surface is carried out silanization with dichlorodimethylsilane and handled to avoid the residual contamination in the liquid introducing process.Coupon (4) is made by 200 microlitre plastic centrifuge tubes, and the bottom all is processed with wide 0.8 millimeter, dark 1 millimeter breach (5).Four coupons (4) are horizontally arranged on one dimension electronic control translation stage (6).Test liquid-hydrogen peroxide the standard solution (7) that adds 100 microlitres in four coupons (4) respectively, reagent solution-luminol solution (8) (7 mMs/liter, pH=10.5), reagent solution-potassium ferricyanide solution (9) (20 mMs/liter), and carrier fluid-deionized water (10).Chemiluminescence detection system on the kapillary adopts photomultiplier (11) as electrooptical device, is placed on kapillary (1) top 0.5 centimeters, and the distance of surveyed area bullet tubule endpiece (3) is 1 centimetre.Kapillary (1) below increases the light quantity that photomultiplier (11) is collected with a luminous mirror (12).Capillary outlet end (3) links to each other with PVC plastic flexible pipe (13) (2 millimeters of internal diameters, 20 centimeter length), and the flexible pipe outlet connects the horizontal sewer pipe (14) by 1 milliliter of injector for medical purpose repacking of a horizontal positioned, forms gravity liquid stream drive system.Be full of deionized water (15) in waste liquid pool and the flexible pipe in advance.By regulating the vertical range of sewer pipe (14), regulate the flow rate of liquid in the kapillary (1) with respect to kapillary entrance point (2).

Fig. 2 adopts the preferred embodiment of the present invention one system to analyze the signal record figure that obtains.Sewer pipe (14) is 6 centimetres with respect to the vertical range of kapillary entrance point (2), 25 degrees centigrade of room temperatures, records flow velocity and is 10.1 and receive liter/second.Kapillary entrance point (2) is soaked by breach (5) carry out the liquid introducing in the coupon (4).The computer control electronic control translation stage carries out the one dimension translation motion, makes kapillary entrance point (2) immerse luminol solution (7), superoxol (8), potassium ferricyanide solution (9) and carrier fluid (10) successively.Wherein in first three solution, respectively stop 0.5 second time (sample size be 5 receive liter), stop 5 second time at carrier fluid (10).Analysis throughput can reach 500 samples/hour.As figure is the record diagram of 1 micromoles per liter hydrogen peroxide standard series, and signal peak height relative standard deviation is 1.0% (n=4).

Embodiment two

Fig. 3 is two a structural representation according to a preferred embodiment of the invention;

Adopt quartz capillary (1) (75 microns of internal diameters; 375 microns of external diameters) constructing system; long 6 centimetres of kapillary (1); the protective layer used sharp pocket knife of kapillary entrance point (2) scrapes off, and kapillary entrance point (2) surface is carried out silanization with dichlorodimethylsilane and handled to avoid the residual contamination in the liquid introducing process.Coupon (4) is made by 200 microlitre plastic centrifuge tubes, and the bottom all is processed with wide 0.8 millimeter, dark 1 millimeter breach (5).10 coupons (4) are horizontally arranged on the coupon array rotating disk (16), are evenly distributed 10 sample hoses (4) on the dish.Pack at interval successively in the coupon fluorescein standard solution (test liquid) and the carrier fluid-deionized water (9) of 100 microlitre variable concentrations, the fluorescein standard solution of variable concentrations comprises 50 nanomoles/liter (17), 100 nanomoles/liter (18), 150 nanomoles/liter (19), 200 nanomoles/liter (20) and 250 nanomoles/liter (21) luciferin solution.Rotating disk (16) by the computer parallel port communication, the time of control rotation direction, angle and stop, realizes that automated fluid is introduced and the accurate control of sampling volume by the segmentation step motor drive.Laser-induced fluorescence detection system (22) (excitation wavelength 473 nanometers) is adopted in the detection of sample in the kapillary.The distance of check point bullet tubule endpiece (3) is 1 centimetre.Capillary outlet end (3) links to each other with PVC plastic flexible pipe (13) (2 millimeters of internal diameters, 20 centimeter length), and the flexible pipe outlet connects the horizontal sewer pipe (14) by 1 milliliter of injector for medical purpose repacking of a horizontal positioned, forms gravity liquid stream drive system.Be full of deionized water (15) in waste liquid pool and the flexible pipe in advance.By regulating the vertical range of sewer pipe (14), regulate the flow rate of liquid in the kapillary (1) with respect to kapillary entrance point (2).

Fig. 4 analyzes the signal record figure that obtains for adopting preferred embodiment of the present invention two system.Sewer pipe (14) is 20 centimetres with respect to the vertical range of kapillary entrance point (2), 25 degrees centigrade of room temperatures, records flow velocity and is 26.4 and receive liter/second.Computer control rotating disk (16) rotates, and makes kapillary entrance point (2) immerse luciferin solution (21), carrier fluid (10), luciferin solution (20), carrier fluid (10), luciferin solution (19), carrier fluid (10), luciferin solution (18), carrier fluid (10), luciferin solution (17) and carrier fluid (10) successively.The residence time is 1 second (sample size be 26.4 receive liter) in each luciferin solution, and the residence time is 7.3 seconds in each carrier fluid (10).Circulation repeats aforesaid operations, obtains 50-250 nanomole/raising of indices directrix curve (R 2=0.9998).Analysis throughput reaches per hour more than 400 samples.

Claims (10)

1, a kind of based on sequential injection analyzer capillaceous, by kapillary, coupon array liquid drawing-in system, the detection system of sample composition in liquid driven system and the kapillary in the kapillary, it is characterized in that capillary outlet is as the liquid driven end, the capillary inlet is as sample, the order valveless leading-in end of reagent and carrier fluid, described capillary inlet end is connected with the coupon outlet of coupon array liquid drawing-in system, coupon array liquid drawing-in system is made up of array and coupon platform that plural coupon constitutes, coupon is fixed on the coupon platform, and capillary outlet is connected with the liquid driven system.
2, according to claim 1ly it is characterized in that the material capillaceous that adopts in the device is for quartzy, perhaps glass, perhaps metal, perhaps macromolecule polymer material based on sequential injection analyzer capillaceous; The capillary channel internal diameter is between 10 nanometers to 5 millimeter; The microcapillary tube wall thickness is in 1 micron to 5 millimeters scope; Capillary pipe length is in 1 millimeter to 10 meters scope.
3, according to claim 1ly it is characterized in that based on sequential injection analyzer capillaceous the coupon exit width is 10 microns to 30 millimeters scopes, the degree of depth is at 1 micron to 5 millimeters; The external diameter scope of coupon is 0.5 millimeter to 10 centimetres.
4, according to claim 1ly it is characterized in that based on sequential injection analyzer capillaceous the coupon platform can carry out one dimension, perhaps two-dimentional, perhaps San Wei translation motion perhaps can be carried out two dimension or three-dimensional rotation, the compound motion that perhaps can carry out translation and rotation.
5, according to claim 1 based on sequential injection analyzer capillaceous, it is characterized in that the liquid driven system is connected with the capillary outlet end, liquid flows to the capillary outlet end from the capillary inlet end, drive system adopts segregation drive power, perhaps electric osmose drives power, perhaps Mechanical Driven power.
6, the described using method of claim 1 based on sequential injection analyzer capillaceous, it is characterized in that, mobile coupon platform drive on it a plurality of coupons in certain sequence successively with the capillary inlet end in contact, the capillary inlet end immerses in the sample liquid in pipe by the coupon import and stops certain hour, under drive systems, make the interior liquid of various sample pipe of certain volume, comprise test liquid and carrier fluid at least, be introduced in certain sequence in the kapillary; Liquid flows to the capillary outlet end from the capillary inlet end with certain flow rate; Near the capillary outlet end, the product of sample or its chemical reaction is detected.
7, the using method based on sequential injection analyzer capillaceous according to claim 6, it is characterized in that the detection method of sample in the kapillary or its reaction product is adopted detecting pattern on the post, perhaps flow out detecting pattern, comprise chemoluminescence method, fluorescence method, photometry, electrochemical method, mass spectroscopy.
8, the using method based on sequential injection analyzer capillaceous according to claim 6 is characterized in that, the residence time scope of kapillary entrance point in coupon liquid is 1 millisecond to 60 minutes.
9, the using method based on sequential injection analyzer capillaceous according to claim 6, the volume range that it is characterized in that being introduced into each liquid capillaceous are 1 to ascend to heaven to 500 microlitres.
10, the using method based on sequential injection analyzer capillaceous according to claim 6 is characterized in that flow rate of liquid in kapillary is 1 a skin liter/minute to 10 ml/min.
CN 200610049830 2006-03-14 2006-03-14 Sequential injection analyzer based on capillaries and utilization thereof CN1818662A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014101575A1 (en) * 2012-12-31 2014-07-03 浙江大学 Application method for automatic micro-droplet array screening system with picoliter precision
CN105092677A (en) * 2014-05-06 2015-11-25 窦晓鸣 Movable electrophoresis chip sample injection method and sample injection device suitable for same

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014101575A1 (en) * 2012-12-31 2014-07-03 浙江大学 Application method for automatic micro-droplet array screening system with picoliter precision
US9804185B2 (en) 2012-12-31 2017-10-31 Zhejiang University Application method for automatic micro droplet array screening system with picoliter scale precision
CN105092677A (en) * 2014-05-06 2015-11-25 窦晓鸣 Movable electrophoresis chip sample injection method and sample injection device suitable for same
CN105092677B (en) * 2014-05-06 2019-02-26 窦晓鸣 Movable electrophoresis chip sample injection method and the sampling device suitable for this method

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