CN203965326U - A kind of Capillary Electrophoresis is with exciting contact fluorescence detection cell - Google Patents
A kind of Capillary Electrophoresis is with exciting contact fluorescence detection cell Download PDFInfo
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- CN203965326U CN203965326U CN201420294729.XU CN201420294729U CN203965326U CN 203965326 U CN203965326 U CN 203965326U CN 201420294729 U CN201420294729 U CN 201420294729U CN 203965326 U CN203965326 U CN 203965326U
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Abstract
The utility model discloses a kind of Capillary Electrophoresis with exciting contact fluorescence detection cell, comprises detection cell body assembly, positioning component and fixation kit.Fluorescence detection cell of the present utility model adopts orthogonal type light channel structure, make to excite, collect optical fiber and liquid light path between two mutual vertical conjugation intersect; The taper reducing tee of adopting quartz glass material connects entrance kapillary, outlet kapillary and excitation fiber, not only realize the physical contact of liquid flow path and excitation fiber, and detection zone still adopts quartzy material, ensure the continuous operation of electroosmotic flow, be applicable to Capillary Electrophoresis-fluoroscopic examination; In addition, designed supporting with it setting element and fixation kit etc., thereby realized the autoregistration of collecting optical fiber and detection zone, made the installation of this detection cell and use more convenient.
Description
Technical field
The utility model relates to a kind of Capillary Electrophoresis contact fluorescence detection cell.
Background technology
Fluoroscopic examination is one of the highest detection technique of current sensitivity, has very high selectivity and the stability of response simultaneously; Capillary Electrophoresis is taking kapillary as split tunnel, ion or charged particle in kapillary by its mobility or/and partition factor difference is carried out efficiently, the technology that separates fast, there is separation efficiency high, the feature such as analysis time is short, and sample size is few.Capillary Electrophoresis-fluorescence detection, combines the advantage of above two kinds of technology, has been widely used in the association areas such as life science, bioengineering, environmental protection, food and medicine monitoring.
In microfluidic analytic system (comprising flow injection, micro column liquid chromatography, Capillary Electrophoresis), on post, detecting is most widely used pattern.One section of polyimide coating of kapillary tail end is removed, and when liquid to be detected is flowed through this region, exciting light sees through capillary wall and is irradiated in liquid, and the fluorescence of generation also sees through capillary wall and is collected.On post, detect and there is minimum extra column effect, make simple, easy to use.But exciting light is reaching before sample, be through two optical interfaces: air-quartz capillary wall, quartz capillary wall-sample solution; Significant refraction coefficient (RI) significant difference (1.0-1.47-1.33), make that light reflects herein, refraction and tube wall inner full-reflection, cylindrical shape capillaceous has more aggravated the optical distortion to excitation beam, the exciting light of reflection is collected optical fiber and receives, thereby causes higher background signal and noise level.
Summary of the invention
For solving problems of the prior art, and for the feature of Capillary Electrophoresis fluorescence detecting system, the utility model provides a kind of Capillary Electrophoresis with exciting contact fluorescence detection cell, and described detection cell comprises detection cell body assembly, positioning component and fixation kit;
Described detection cell body assembly comprises:
T-shaped three-way pipe, its main pipeline and branch pipe(tube) inwall are respectively equipped with taper reducer, and the bigger diameter end of taper reducer is all away from T-shaped three-way pipe center,
Insert and terminate in respectively entrance kapillary and the outlet kapillary of taper reducer from main pipeline openend, and
Insert and terminate in the excitation fiber of taper reducer from branch pipe(tube) openend;
The region being enclosed by entrance kapillary, outlet kapillary and excitation fiber in described T-shaped three-way pipe is detection zone;
Described positioning component comprises:
Platform body, which is provided with the T-shaped V groove matching with T-shaped three-way pipe profile,
Be provided with through hole I perpendicular to platform surface, this through hole I is positioned at the main channel of V groove and the intersection point place of a conduit, and through hole I expands to the one end away from platform in succession for coaxial circular cylindrical cavity I and cavity II,
The elastomeric element in the tool hole matching with cavity I,
The securing member in the tool hole matching with cavity II,
Through hole III on through hole II and securing member on through hole I, elastomeric element joins and becomes collection optical-fibre channel, the fixing optical fiber of collecting in it;
For detection cell body assembly is fixed on the fixation kit on platform body and is provided with form corresponding to position, detection zone.
The technical solution of the utility model possesses following outstanding feature:
1, ensureing, under the continuous prerequisite of electroosmotic flow, to have realized the physical contact of excitation fiber and liquid flow path, eliminated the interference of stray light that on post, detection brings due to capillary wall optical interface.
2, adopt taper reducing to be connected with entrance, outlet kapillary and excitation fiber detection zone, ensureing to be coated with under the operability prerequisite of glue bond, realized at the diameter of interface minimum and having changed, thereby reduce extra column effect.
3, designed and matching used retaining element of detection cell body etc., this retaining element adopts V-shape groove, can play hole from the T of groove font intersection central straight, as the pilot hole of collecting optical fiber, thereby process and can realize the auto-alignment of collecting optical fiber and surveyed area by standard machinery.
4, detection cell body and collection optical fiber all adopt silicagel pad distortion to be fixed, and have good anti-seismic performance.
5, the design of fixation kit ensures that detection zone and important junction all can be in visual states, be convenient to install and trouble hunting: when installation: detection cell body is originally as transparent quartz glass material, can see through glass, immediate observation is to collecting the plane relative position (whether the end face of collecting optical fiber aimed at detection zone) of optical fiber with detection zone; When trouble hunting, 1) can open black out cover, need not move any one parts, can be observed kapillary and whether normally bond with being connected of detection cell body with detection cell body, excitation fiber; In Capillary Electrophoresis process, if having bubble in damping fluid and enter into liquid flow path, may cause the fracture of electroosmotic flow, easily produce this phenomenon at interface especially, visually support user under need not the prerequisite of dismounting, observe directly this phenomenon, and deal with in time.
To sum up, fluorescence detection cell of the present utility model adopts orthogonal type light channel structure, make to excite, collect optical fiber and liquid light path between two mutual vertical conjugation intersect; The taper reducing tee of adopting quartz glass material connects entrance kapillary, outlet kapillary and excitation fiber, not only realize the physical contact of liquid flow path and excitation fiber, and detection zone still adopts quartzy material, ensure the continuous operation of electroosmotic flow, be applicable to Capillary Electrophoresis-fluoroscopic examination; In addition, designed supporting with it setting element and fixation kit etc., thereby realized the autoregistration of collecting optical fiber and detection zone, made the installation of this detection cell and use more convenient.
Brief description of the drawings
The utility model accompanying drawing 9 width:
Fig. 1 is T-shaped three-way pipe structure cut-open view in detection cell body assembly (5 times of amplification);
Fig. 2 is detection cell body assembly 100 structure cut-open views (5 times of amplifications);
Fig. 3 is the front view of positioning component 200;
Fig. 4 is the rear perspective view of positioning component 200;
Fig. 5 is the rear view of positioning component 200;
Fig. 6 is the A-A' cut-open view of Fig. 5;
Fig. 7 is the front view that excites contact fluorescence detection cell for Capillary Electrophoresis of assembling, partly cut-away;
Fig. 8 is the B-B' cut-open view (not containing fixation kit) of Fig. 7;
Fig. 9 is that detection cell of the present utility model is applied to the Capillary Electrophoresis spectrogram that Capillary Electrophoresis-detector of fluorescence induced by light-emitting diode analysis ivermectin obtains.
In figure:
100, detection cell body assembly; 110, T-shaped three-way pipe; 111,112, main pipeline; 131, branch pipe(tube); 112,122,132, taper reducer; 113, entrance kapillary; 123, outlet kapillary; 133, excitation fiber; 140, detection zone;
200, positioning component; 210, platform body; 211, V groove; 212, through hole I; 213, cavity I; 214, cavity II; 220, elastomeric element; 230, securing member; 240, collect optical fiber;
300, fixation kit.
Embodiment
By reference to the accompanying drawings to Capillary Electrophoresis of the present utility model with exciting contact fluorescence detection cell to be described further.
Capillary Electrophoresis of the present utility model is with exciting contact fluorescence detection cell by three module compositions, be respectively detection cell body assembly 100, positioning component 200 and fixation kit 300, detection cell body assembly 100 wherein provides the flow passage of detected material, and the functional areas of fluorescence excitation, fluorescent collecting, positioning component 200 combines with fixation kit 300 for detection of the fixing and calibration of pond body assembly 100.
Described detection cell body assembly 100 comprises T-shaped three-way pipe 110, entrance kapillary 113, outlet kapillary 123 and excitation fiber 133.From main pipeline 111,121, (two openends that are two shoulder pipes of T-shaped three-way pipe insert, and excitation fiber 133 is inserted from branch pipe(tube) 131 openends of T-shaped three-way pipe 110 respectively for entrance kapillary 113 and outlet kapillary 123.
Described T-shaped three-way pipe 110 is preferably made up of quartz glass, and main pipeline 111,121 and branch pipe(tube) 131 inwalls are therein respectively equipped with taper reducer 112,122 and 132, and the bigger diameter end of taper reducer is all away from T-shaped three-way pipe 110 centers.Preferably in described T-shaped three-way pipe 110, the reducing angle of taper reducer is 20～50 degree.In the utility model, reducing angle refers to half cone apex angle, and this angle is 30～45 degree preferably.
The external diameter of above-mentioned entrance kapillary 113 is between the bigger diameter end diameter and miner diameter end diameter of main pipeline 111 inner conical reducers 112, this design ensures that entrance kapillary 113 can insert and terminate in taper reducer 112 from main pipeline 111 openends, equally, outlet kapillary 123 can insert and terminate in taper reducer 122 from another main pipeline 121 openends, and taper reducer 132 is inserted and terminated in to excitation fiber 133 from branch pipe(tube) 131 openends.As reality can application mode, when detection cell body assembly 100 combines, can first coat curing glue at the insertion end outer wall of entrance kapillary 113, outlet kapillary 123 and excitation fiber 133, then they are inserted to the corresponding pipeline depths as far as possible of T-shaped three-way pipe 110, when the section of having smeared curing glue is hampered by taper reducer and cannot continues to insert, standing assembly or high-temperature baking are made to solidify glue fully curing, thereby complete the assembling of detection cell body assembly 100.
After assembling, the region being enclosed by entrance kapillary 113, outlet kapillary 123 and excitation fiber 133 in described T-shaped three-way pipe 110 becomes detection zone 140.
Described positioning component 200 is preferably made up of the aluminium oxide that turning black or stainless steel.This positioning component 200 comprises platform body 210, which is provided with the T-shaped V groove 211 matching with T-shaped three-way pipe 110 profiles, be provided with through hole I212 perpendicular to platform body 210 surfaces, this through hole I212 is positioned at the main channel of V groove 211 and the intersection point place of a conduit, through hole I212 expands to the one end away from platform 210 in succession for coaxial circular cylindrical cavity I213 and cavity II214, positioning component 200 also comprises the elastomeric element 220 in the tool hole matching with cavity I213, the securing member 230 in the tool hole matching with cavity II214 and collect optical fiber 240, described collection optical fiber 240 is accommodated in by through hole I212, through hole III on through hole II and securing member 230 on elastomeric element 220 joins to becoming and collects in optical-fibre channel.
As reality can application mode, described cavity II214 inwall is provided with internal thread, securing member 230 is provided with the external thread matching with it; Described cavity I213 and elastomeric element 220 are right cylinder, and the former degree of depth is slightly less than the height of the elastomeric element 220 in tool hole.When assembling, above-mentioned collection optical fiber 240 is passed to through hole III, through hole II and through hole I successively, make to collect end face and detection cell body assembly 100 physical contacts of optical fiber, cavity II214 is threaded with securing member 230, in the process screwing, thereby elastomeric element 220 compressive deformations are realized collecting the fixing of optical fiber 240.
Described fixation kit 300 is for being fixed to platform body 210 by detection cell body assembly 100, and fixation kit 300 is also made up of the aluminium oxide that turning black or stainless steel, preferably material identical with positioning component 200.
In specific embodiment, described fixation kit 300 can be independently by 3 and the fixed component of platform body 210 assembly and connections form, three fixed components respectively three branch pipe(tube)s of T-shaped three-way pipe 110 are pressed and fixed in the V groove 211 on platform body 210 but do not cover the detection zone 140 of T-shaped three-way pipe 110.Described fixed component is aluminium oxide or the stainless steel component that is provided with the blackout of screw, and described platform body 210 is identical with fixed component material, which is provided with the screw matching with fixed component, by connecting screw rod, fixed component is fixedly connected with platform body 210.
Described fixation kit (300) also comprises the elastic component matching with it, elastic component is located between fixation kit (300) and platform body (210), it is pressed on fixation kit (300) and is produced deformation, thereby the detection cell body assembly (100) in platform body (210) V groove (211) is fixing.Under assembled state, described detection cell body assembly 100 is positioned at T-shaped V groove 211 set on the platform body 210 of positioning component 200, preferably collaboration elastic component is fixed, one of concrete selection of described elastomeric element is that thickness is the silicagel pad of 2～10mm, shape is consistent with fixation kit, when assembling, be installed between fixation kit 300 and platform body 210 the local detection cell body assembly 100 that covers; In the time that fixation kit 300 is exerted pressure to it, produce deformation as the silicagel pad of elastic component, thereby by fixing the detection cell body assembly in platform body 210V groove 211 100.
In concrete embodiment, on fixation kit 300, be provided with form corresponding to 140 positions, detection zone, this is for the ease of assembling and visualized operation when repair and maintenance.From processing, reality is installed, described fixation kit 300 is designed to 3 can be independently and the fixed component of platform body 210 assembly and connections composition, three fixed components respectively three branch pipe(tube)s of T-shaped three-way pipe 110 are pressed and fixed in the V groove 211 on platform body 210 but do not cover the detection zone 140 of T-shaped three-way pipe 110.Each fixed component is provided with screw on (comprising silicagel pad), described platform body 210 be provided with fixed component on the screw that matches of screw, connecting screw rod is fixedly connected with fixed component (comprising silicagel pad) with platform body 210.
Certainly, the requirement to environment in detecting for reality, Capillary Electrophoresis of the present utility model is with exciting contact fluorescence detection cell also to comprise lightshade cover, and described lightshade cover covers detection zone 140 completely, preferably covers platform body 210 completely.
Embodiment 1
A kind of Capillary Electrophoresis, with exciting contact fluorescence detection cell, assembles according to following step and parameter:
1, assembling detection cell body assembly 100, as shown in attached Fig. 1 and 2:
The transparent T-shaped three-way pipe 110 that quartz glass is made, its main pipeline 111,121 and branch pipe(tube) 131 inwalls are respectively equipped with taper reducer 112,122 and 132, and the bigger diameter end of taper reducer is all away from T-shaped three-way pipe 110 centers; 3 millimeters of T-shaped three-way pipe 110 external diameters, 500 microns of internal diameters, after angle 45 is spent taper reducer, diameter is 100 microns.Be that 365 microns, internal diameter are that kapillary one port outer wall of 100 microns is smeared and solidified after glue by external diameter, insert from main pipeline 111,121 openends respectively, until taper reducer 112,122 is arrived in termination; One port outer wall of excitation fiber 133 is smeared and is solidified after glue, inserts until taper reducer 132 is arrived in termination from branch pipe(tube) 131 openends; What in T-shaped three-way pipe 110, enclosed by entrance kapillary 113, outlet kapillary 123 and excitation fiber 133 forms detection zone 140.
2, with aluminium oxide casting positioning component 200 and the fixation kit 300 of turning black
As accompanying drawing 3～6, the platform body 210 of described positioning component 200 is provided with the T-shaped V groove 211 matching with T-shaped three-way pipe 110 profiles, and the V-shape angle 60 of described V groove is spent, 4 millimeters of groove depths.The main channel of V groove 211 arranges through hole I212 with the location, intersection point place of conduit, and this through hole expands to the one end away from platform 210 in succession for coaxial circular cylindrical cavity I213 and cavity II214, the former diameter 5mm, and dark 4mm, the latter is provided with internal thread.On platform body 210, be also provided with 6 of screws for being connected with fixation kit 300.Meanwhile, positioning component 200 is also equipped with the elastomeric element 220 in the tool hole matching with cavity I213, i.e. cylindrical tool hole silica gel part (diameter 4.5mm, height 4.5mm), and the externally threaded securing member 230 of tool hole, tool matching with cavity II214.
Described fixation kit 300 be 3 can be independently and the fixed component of platform body 210 assembly and connections composition, three fixed components respectively three branch pipe(tube)s of T-shaped three-way pipe 110 are pressed and fixed in the V groove 211 on platform body 210 but do not cover the detection zone 140 of T-shaped three-way pipe 110.Each fixed component is provided with 2 screws on (comprising silicagel pad), the screw being provided with platform body 210 matches, and can fixed component (comprising silicagel pad) be fixedly connected with platform body 210 by connecting screw rod.Fixation kit 300 also comprises that thickness that 3 shapes and fixed component match is about the silicagel pad (as accompanying drawing 7) of 3mm.
When overall package, the detection cell body assembly 100 that assembling is completed is put into T-shaped V groove 211 set on platform body 210, will be installed on as the silicagel pad of fixed component between the member and platform body 210 of fixation kit 300, the local detection cell body assembly 100 that covers; In the time that fixation kit 300 is exerted pressure to it, produce deformation as the silicagel pad of elastic component, thereby by fixing the detection cell body assembly in platform body 210V groove 211 100.When the shape setting of fixed component (comprising silicagel pad) makes it be combined into fixation kit 300 and play a role, form enough large form, thereby make detection zone 140 visual (accompanying drawing 7).Under visibility status detection zone (140 is also transparent), to collect optical fiber 240 successively through through hole III, through hole II and through hole I, make to collect end face and detection cell body assembly 100 physical contacts of optical fiber, cavity II214 is threaded with securing member 230, in the process screwing, thereby elastomeric element 220 compressive deformations are realized fixing (accompanying drawing 8) to collecting optical fiber 240.
Finally, use lightshade cover cover completely platform body and with platform body, when maintenance, remove.
Embodiment 2
Set up and excite contact fluorescence detection cell according to embodiment 1, and apply this detection cell assembling detector of fluorescence induced by light-emitting diode, be applied to Capillary Electrophoresis and detect Ivermectin HCL content in commercially available grass carp.
Assembling scheme: adopt the light emitting diode of centre wavelength 365nM, band flow-through interference filter BP365nm is to the light source processing that filters, the 5V constant pressure source 100 Ω resistance of contacting are that light emitting diode is powered; Conduct exciting light by excitation fiber and be irradiated to detection zone.Carry out phosphor collection in the direction vertical with exciting light, through fiber optic conduction, eliminate background interference light by fluorescent optical filter (BP475nm), finally enter photomultiplier and be converted to electric signal and output to workstation.
Test condition: buffer solution: buffer solution: Na2B4O7 (10mM, pH8.0); Fused quartz kapillary: 365 μ m (external diameter) × 100 μ m (internal diameter) × 55/45cm (overall length/effectively); Separation voltage: 20kV.
Test result: quantitative by external standard method, record in commercially available grass carp, ivermectin content is 4.5 μ g/kg; " the how residual mensuration high performance liquid chromatography of Avermectin and ivermectin in GB/T29695-2013 national food safety standard aquatic products " measures numerical value is 4.3 μ g/kg.
Claims (9)
1. Capillary Electrophoresis, with exciting a contact fluorescence detection cell, is characterized in that: comprise detection cell body assembly (100), positioning component (200) and fixation kit (300);
Described detection cell body assembly (100) comprising:
T-shaped three-way pipe (110), its main pipeline (111,121) and branch pipe(tube) (131) inwall are respectively equipped with taper reducer (112,122 and 132), the bigger diameter end of taper reducer is all away from T-shaped three-way pipe (110) center
Insert and terminate in respectively entrance kapillary (113) and the outlet kapillary (123) of taper reducer (112,122) from main pipeline (111,121) openend, and
Insert and terminate in the excitation fiber (133) of taper reducer (132) from branch pipe(tube) (131) openend;
The region being enclosed by entrance kapillary (113), outlet kapillary (123) and excitation fiber (133) in described T-shaped three-way pipe (110) is detection zone (140);
Described positioning component (200) comprising:
Platform body (210), which is provided with the T-shaped V groove (211) matching with T-shaped three-way pipe (110) profile,
Be provided with through hole I (212) perpendicular to platform body (210) surface, this through hole I (212) is positioned at the main channel of V groove (211) and the intersection point place of a conduit, through hole I (212) is in succession expanding away from one end of platform (210) for coaxial circular cylindrical cavity I (213) and cavity II (214)
The elastomeric element (220) in the tool hole matching with cavity I (213),
The securing member (230) in the tool hole matching with cavity II (214),
Through hole III on through hole II and securing member (230) on through hole I (212), elastomeric element (220) joins and becomes collection optical-fibre channel, the fixing optical fiber (240) of collecting in it;
Above be provided with form corresponding to position, detection zone (140) for the fixation kit (300) detection cell body assembly (100) being fixed on platform body (210).
2. Capillary Electrophoresis according to claim 1 is with exciting contact fluorescence detection cell, it is characterized in that: described fixation kit (300) can be independently by 3 and the fixed component of platform body (210) assembly and connection form, three fixed components respectively three branch pipe(tube)s of T-shaped three-way pipe (110) are pressed and fixed in the V groove (211) on platform body (210) but do not cover the detection zone (140) of T-shaped three-way pipe (110).
3. Capillary Electrophoresis according to claim 2 is with exciting contact fluorescence detection cell, it is characterized in that: described fixed component is aluminium oxide or the stainless steel component that is provided with the blackout of screw, described platform body (210) is identical with fixed component material, which is provided with the screw matching with fixed component, by connecting screw rod, fixed component is fixedly connected with platform body (210).
According to the Capillary Electrophoresis described in claim 1,2 or 3 with exciting contact fluorescence detection cell, it is characterized in that: described fixation kit (300) also comprises the elastic component matching with it, elastic component is located between fixation kit (300) and platform body (210), it is pressed on fixation kit (300) and is produced deformation, thereby the detection cell body assembly (100) in platform body (210) V groove (211) is fixing.
5. Capillary Electrophoresis according to claim 4, with exciting contact fluorescence detection cell, is characterized in that: described elastic component is that thickness is the silicagel pad of 2～10mm.
6. Capillary Electrophoresis according to claim 1, with exciting contact fluorescence detection cell, is characterized in that: in described T-shaped three-way pipe (110), the reducing angle of taper reducer is 20～50 degree.
7. Capillary Electrophoresis according to claim 1, with exciting contact fluorescence detection cell, is characterized in that: described T-shaped three-way pipe (110) is quartz glass.
8. Capillary Electrophoresis according to claim 1, with exciting contact fluorescence detection cell, is characterized in that: described cavity II (214) inwall is provided with internal thread, and securing member (230) is provided with the external thread matching with it.
9. Capillary Electrophoresis according to claim 1, with exciting contact fluorescence detection cell, is characterized in that: also comprise the lightshade cover that covers detection zone (140) at least completely.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109870411A (en) * | 2017-12-04 | 2019-06-11 | 中国科学院大连化学物理研究所 | A kind of adjustable calibrating --- refractive power compensates fluorescence detection cell on column |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109870411A (en) * | 2017-12-04 | 2019-06-11 | 中国科学院大连化学物理研究所 | A kind of adjustable calibrating --- refractive power compensates fluorescence detection cell on column |
| CN109870411B (en) * | 2017-12-04 | 2021-07-06 | 中国科学院大连化学物理研究所 | Adjustable calibration refraction compensation column fluorescence detection pool |
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