CN102539512B - Multichannel capillary electrophoresis method for detecting glycosylated hemoglobin HbA1c in multiple samples simultaneously - Google Patents
Multichannel capillary electrophoresis method for detecting glycosylated hemoglobin HbA1c in multiple samples simultaneously Download PDFInfo
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Abstract
The invention relates to a multichannel capillary electrophoresis method for detecting glycosylated hemoglobin HbA1c in multiple samples simultaneously. According to the multichannel capillary electrophoresis method, the glycosylated hemoglobin HbA1c in multiple samples is detected simultaneously through an array capillary and an optic array detection device. The glycosylated hemoglobin HbA1c in multiple blood samples can be analyzed and detected simultaneously, and results are automatically analyzed to form a report.
Description
Technical field
This invention relates to a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously.
Background technology
To the detection of glycosylated hemoglobin, all exist detection flux low at present, the situation that cost is high or accuracy is low always.
For example the accuracy of immunization is poor, and cost is also higher; The accuracy of ion-exchange chromatography method is very high, but has no idea to realize high flux test, and experimental cost is also very high, and common patient is difficult to accept.The present invention is intended to set up a set of high flux, high-speed, can detect the capillary array electrophoresis method of a plurality of samples, the requirement detecting to meet glycosylated hemoglobin simultaneously.
Multiple-pass capillary tube electrophoresis technology has been successfully applied to DNA sequencing system, but multiple-pass capillary tube electrophoresis system is detected and there is no at home report for glycosylated hemoglobin HbAlc index; Domestic at present, the existing report that single capillary electrophoresis method is detected for glycosylated hemoglobin HbAlc, but the method flux is too low, cannot meet the requirement that great amount of samples detects.This method combines the multiple-pass capillary tube electrophoresis technology of using in research work and single track Capillary Electrophoresis glycosylated hemoglobin detection technique, both solved the difficult problem that glycosylated hemoglobin HbAlc detects, greatly improve again the flux and the speed that detect, met the requirement of testing laboratory.
Summary of the invention
The present invention measures high flux and Fast Measurement for glycosylated hemoglobin HbAlc relative content.The present invention uses the array of many capillaries can the glycosylated hemoglobin HbAlc in a plurality of blood samples be analyzed and be detected simultaneously as split tunnel, and automatically result is analyzed, and generates report.
The invention provides a kind ofly for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, use array capillary and optical arrays pick-up unit a plurality of samples to be carried out to glycosylated hemoglobin HbAlc detection simultaneously.
Preferably, described detection method comprises the following steps:
1) dissociating buffer is filled with in each kapillary in array capillary as isolating environment;
2) each sample is joined simultaneously to each front end capillaceous in array capillary;
3) after sample enters each kapillary front end in capillary array, the two ends of array capillary are replaced by dissociating buffer;
4) at kapillary two ends, apply voltage as separated driving force;
5) by detection light source and optical arrays detecting device, the sample in kapillary is carried out to glycosylated hemoglobin HbAlc detection;
6) according to step 5) gained detects the content that data calculate the glycosylated hemoglobin HbAlc in each sample.
Preferred, capillary inner diameter 20-100 micron in described capillary array, effectively separation length 10-50 centimetre.
Preferred, in described capillary array, quantity capillaceous is 4-48 root, and described kapillary is without coating quartz capillary or the cated neutral quartz capillary of inwall.
Preferred, internal diameter and length that described kapillary is every are all consistent.
Further, described 4-48 capillary forms capillary array as split tunnel, can carry out electrophoretic separation and detection to 4-48 sample while sample introduction.
Preferred, described each front portion capillaceous is parallel to each other, and end set is to a place.
Preferred, in step 1) front, at the capillary tube inner wall that forms described array capillary, prepare coating.
Preferred, described coating is the duplex coating that polycation coating and polyanion coating form.
Preferred, the method for preparing described duplex coating is: first by said polycation solution, rinse kapillary, then rinse kapillary with polyanion solution.
Preferred, before preparation coating, use the kapillary in strong acid and/or strong base solution pair array kapillary to process.
Preferred, kapillary in described strong acid and/or strong base solution pair array kapillary is processed, polycation and polyanion solution rinse kapillary, dissociating buffer is filled with process capillaceous and all by pressure, completes, described strong acid and strong base solution, polycation and polyanion solution, dissociating buffer all enter from the end of capillary array, from front end, are discharged to waste liquid pool.
Preferred, described strong acid solution is selected from hydrochloride buffer or nitric acid damping fluid, and the concentration of described strong acid solution is 0.01-1mol/L.
Preferred, described strong base solution is selected from sodium hydrate aqueous solution or lithium hydroxide aqueous solution, and the concentration of described strong base solution is 0.01-1mol/L.
Preferred, described said polycation solution is the aqueous solution that contains polycation, is selected from polybrene aqueous solution, diallyl dimethyl ammoniumchloride aqueous solution, albumin aqueous solution, and its concentration is 0.001-1wt%.
Preferred, described polyanion solution is the aqueous solution that contains polyanion, is selected from polyacrylic acid aqueous solution, kayexalate aqueous solution, chondroitin sulfate aqueous solution, and its concentration is 0.001-1wt%.
Because uncoated kapillary can be adsorbed onto sample inside surface capillaceous, conventionally can not isolate the peak of glycosylated hemoglobin HbAlc, thereby can not go out peak.So the present invention uses polyanion coating and polycation coating to adsorb at capillary tube inner wall to reach the albumen of controlling in blood sample on without coatings capillary pipe tube wall, and accelerates the object that peak speed of glycosylated hemoglobin.Multiple-pass capillary tube electrophoresis system involved in the present invention is used the capillary array forming without coatings capillary pipe as split tunnel, to reach reduction testing cost, keeps capillaceous compared with the object of long life simultaneously.But multiple-pass capillary tube electrophoresis system involved in the present invention also can directly be used the cated capillary array of inwall as split tunnel (such as the kapillary that can select the coatings such as polyacrylamide, polyvinyl alcohol (PVA), polypyrrole alkane ketone), also can on the cated kapillary of inwall, re-use polyanion coating or polycation coating.
Preferred, the pH value of described dissociating buffer is 3-5.
Preferred, described step 2) in each sample in, contain electroendosmotic flow marker.
Preferred, the appearance time of described electroendosmotic flow marker of take is benchmark, draws glycosylated hemoglobin and the corresponding peak of non-glycosylated hemoglobin, and peak value is carried out to the content that integration obtains glycosylated hemoglobin in sample.
Preferred, the appearance time of described electroendosmotic flow marker of take is benchmark, draw glycosylated hemoglobin and the corresponding peak of non-glycosylated hemoglobin, the peak value at glycosylated hemoglobin and the corresponding peak of non-glycosylated hemoglobin is carried out obtaining after integration also calculates to the content of glycosylated hemoglobin in sample.
Preferred, the formula of described calculating is: glycosylated hemoglobin (mol%) content=glycosylated hemoglobin peak value/(glycosylated hemoglobin peak value+non-glycosylated hemoglobin peak value) * 100%.
Preferred, described step 3) in, electroendosmotic flow marker is dimethyl sulfoxide; The volume ratio of described dimethyl sulfoxide and sample is 1: 1-30.
Preferred, described step 3) sample in comprises blood sample and hemolytic agent, and the volume ratio of described blood sample and hemolytic agent is 1: 1-3, described hemolytic agent can be commercially available commercialization hemolytic agent.
Preferred, described step 3) in, to join the method for each front end capillaceous in capillary array be voltage system or pressure mode to each sample simultaneously.
Preferred, the sample size of described sample is 1-100nL.
Preferred, described step 4) in, execute alive mode for electrode being inserted in the dissociating buffer at kapillary two ends or being directly connected on kapillary two ends.
Preferred, described step 4) voltage applying in is constant dc, and U=10-30kV.
Preferred, described step 5) light source in is that wavelength is ultraviolet or the visible light source of 180-600nm.
Preferred, described ultraviolet or visible light source are Halogen lamp LED or LED light source, described optical arrays detecting device comprises a plurality of photodetectors, and described photodetector mates one by one with kapillary, and described optical arrays detecting device is the optical arrays detecting device that PMT or CCD detecting device form.
More have choosing, described peak value is uv absorption peak value.
Because the two ends of capillary array have applied High Level DC Voltage (10-30kV), under High Level DC Voltage effect, blood protein in every capillary can start electrophoresis under electric field force effect, but due to electrophoretic velocity difference, glycosylated hemoglobin HbAlc can separate with other kind of proteinoid.In described capillary array, the afterbody of parallel-segment capillaceous is provided with transparent detection window, and described effective separation length is that capillary inlet is to the distance between detection window.Because each effective separation length capillaceous all equates, so the deposition condition of each sample is basic identical, that is to say that same composition in each sample arrives time of transparent detection window basic identical.The light signal that detection light source is sent is perpendicular to detection window, and arrive the photodetector corresponding with this detection window by detection window, when electroendosmotic flow marker, glycosylated hemoglobin HbAlc and other kind of albuminoid pass through transparent detection window, photodetector in the optical arrays detecting device corresponding with this detection window can be recorded the response of each component and detection data are sent to signal analysis unit, and signal analysis unit draws the absorption peak peak value of each component after the data obtained is processed.
Accompanying drawing explanation
Fig. 1 is principle of the invention schematic diagram (take single capillary as example)
Fig. 2 is testing result schematic diagram of the present invention (take single capillary as example)
Embodiment
Multiple-pass capillary tube electrophoresis system involved in the present invention can be used the capillary array forming without coatings capillary pipe as split tunnel, to reach reduction testing cost, keeps capillaceous compared with the object of long life simultaneously.Described capillary array comprises 4-48 root elastic quartz capillary tube (without coating, internal diameter 20-100 micron, length 10-50cm), and each front portion capillaceous is parallel to each other, and end set is to a place.The present invention also simultaneously forms polycation and polyanion coating at capillary tube inner wall, with what accelerate glycosylated hemoglobin, goes out peak speed.In described sample, the content of each component detects by ultraviolet/visible light optical arrays detecting device.
The principle of the principle of the invention (be take single capillary as example) as shown in Figure 1.Principle of the invention schematic diagram, comprises sample injection unit, optical detection unit and signal analysis unit 8 as shown in Figure 1, and described sample injection unit matches with described optical detection unit, and described optical detection unit is electrically connected to signal analysis unit 8.
Further, described optical detection unit comprises detection light source 7, optical arrays detecting device 3 and array capillary bundle, and described array capillary bundle matches with described sample injection unit, and detection light source 7 and optical arrays detecting device 3 match with described array capillary bundle.
Further, the inwall that forms the kapillary 9 of described array capillary bundle is provided with polycation coating and polyanion coating.
Further, described array capillary bundle consists of 4-48 capillary 9.
Further, between the kapillary 9 of described forming array capillary bundle, be parallel to each other, one end of kapillary 9 is that front end 1, the other end are end 2; And end 2 is collected to a place.
Further, the kapillary 9 of described forming array capillary bundle is provided with transparent detection window 10, and transparent detection window 10 is positioned at the parallel portion of kapillary 9, matches with detection light source 7 in the position of transparent window 10.
Further, the kapillary 9 of described forming array capillary bundle is internal diameter 20-100 micron.
Below in conjunction with specific embodiment, further set forth the present invention, should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.
Embodiment 1:
Select 10 elastic quartz capillary tubes (20 microns of internal diameters, long 30cm) and metal electrode to form capillary array.At capillary array rear portion, be provided with transparent detection window, use ultraviolet light to carry out quantitative and qualitative analysis detection to the separated protein component obtaining in kapillary.
After elastic quartz capillary tube 0.1mol/L NaOH aqueous solution is processed, more successively with polycation (diallyl dimethyl ammoniumchloride, 0.5wt%) and polyanion (chondroitin sulfate, 0.5wt%) aqueous solution flushing kapillary, obtains duplex coating.Subsequently the citric acid solution of pH=4 is filled with to kapillary as isolating environment.Each flushing capillaceous, coating procedure all complete by pressure above, and enter (endpiece from the end of capillary array, one end with respect to injection port), from front end, be discharged to waste liquid pool, the solution tank of described NaOH aqueous solution, polycation aqueous solution, polyanion aqueous solution and citric acid solution is controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
The inlet end capillaceous of capillary array is inserted in array sample groove one to one, 50nL sample is entered to front end capillaceous in capillary array with voltage or pressure mode simultaneously, and the allotment volume ratio of described sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 3.
Kapillary two ends are connected for dissociating buffer system the constant DC voltage that adds 10-30kV on kapillary are as separated driving force, with drive various albumen in sample with friction speed by detection window position.Described array sample groove and dissociating buffer system are controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
Each protein component flowing through in kapillary detects by ultraviolet light array detector, forms and detects collection of illustrative plates, and be recorded in software.
Label using dimethyl sulfoxide as electroosmotic flow, the appearance time at other each peak and dimethyl sulfoxide as a reference, are confirmed the protein component of each peak representative, its testing result (be take single capillary as example) as shown in Figure 2.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out to integration, obtain the percentage of glycosylated hemoglobin HbAlc.
Embodiment 2:
Select 4 elastic quartz capillary tubes (50 microns of internal diameters, long 50cm) and metal electrode to form capillary array.At capillary array rear portion, be provided with transparent detection window, use ultraviolet light to carry out quantitative and qualitative analysis detection to the separated protein component obtaining in kapillary.
After elastic quartz capillary tube 1mol/L KOH aqueous solution is processed, more successively with polycation (polybrene aqueous solution, 1wt%) and polyanion (polyacrylic acid aqueous solution, 0.001wt%) aqueous solution flushing kapillary, obtains duplex coating.Subsequently the citric acid solution of pH=5 is filled with to kapillary as isolating environment.Each flushing capillaceous, coating procedure all complete by pressure above, and enter (endpiece from the end of capillary array, one end with respect to injection port), from front end, be discharged to waste liquid pool, the solution tank of described NaOH aqueous solution, polycation aqueous solution, polyanion aqueous solution and citric acid solution is controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
The inlet end capillaceous of capillary array is inserted in array sample groove one to one, 100nL sample is entered to front end capillaceous in capillary array with voltage or pressure mode simultaneously, and the allotment volume ratio of described sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 9: 21.
Kapillary two ends are connected for dissociating buffer system the constant DC voltage that adds 10-30kV on kapillary are as separated driving force, with drive various albumen in sample with friction speed by detection window position.Described array sample groove and dissociating buffer system are controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
Each protein component flowing through in kapillary detects by ultraviolet light array detector, forms and detects collection of illustrative plates, and be recorded in software.
Label using dimethyl sulfoxide as electroosmotic flow, the appearance time at other each peak and dimethyl sulfoxide as a reference, are confirmed the protein component of each peak representative.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out to integration, and the percentage that obtains glycosylated hemoglobin HbAlc is substantially the same manner as Example 1.
Embodiment 3:
Select 30 elastic quartz capillary tubes (75 microns of internal diameters, long 10cm) and metal electrode to form capillary array.At capillary array rear portion, be provided with transparent detection window, use visible ray to carry out quantitative and qualitative analysis detection to the separated protein component obtaining in kapillary.
After elastic quartz capillary tube 0.1mol/L KOH aqueous solution is processed, more successively with polycation (albumin aqueous solution, 0.1wt%) and polyanion (chondroitin sulfate, 0.05wt%) aqueous solution flushing kapillary, obtains duplex coating.Subsequently the citric acid solution of pH=3 is filled with to kapillary as isolating environment.Each flushing capillaceous, coating procedure all complete by pressure above, and enter (endpiece from the end of capillary array, one end with respect to injection port), from front end, be discharged to waste liquid pool, the solution tank of described NaOH aqueous solution, polycation aqueous solution, polyanion aqueous solution and citric acid solution is controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
The inlet end capillaceous of capillary array is inserted in array sample groove one to one, 10nL sample is entered to front end capillaceous in capillary array with voltage or pressure mode simultaneously, and the allotment volume ratio of described sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 10: 10.
Kapillary two ends are connected for dissociating buffer system the constant DC voltage that adds 10-30kV on kapillary are as separated driving force, with drive various albumen in sample with friction speed by detection window position.Described array sample groove and dissociating buffer system are controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
Each protein component flowing through in kapillary detects by visible ray optical arrays detecting device, forms and detects collection of illustrative plates, and be recorded in software.
Label using dimethyl sulfoxide as electroosmotic flow, the appearance time at other each peak and dimethyl sulfoxide as a reference, are confirmed the protein component of each peak representative.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out to integration, and the percentage that obtains glycosylated hemoglobin HbAlc is substantially the same manner as Example 1.
Embodiment 4:
Select 48 elastic quartz capillary tubes (100 microns of internal diameters, long 40cm) and metal electrode to form capillary array.At capillary array rear portion, be provided with transparent detection window, use ultraviolet light to carry out quantitative and qualitative analysis detection to the separated protein component obtaining in kapillary.
After elastic quartz capillary tube 0.01mol/L NaOH aqueous solution is processed, again successively by polycation (diallyl dimethyl ammoniumchloride, 0.001wt%) and polyanion (kayexalate aqueous solution, 1wt%) aqueous solution is rinsed kapillary, obtains duplex coating.Subsequently the citric acid solution of pH=5 is filled with to kapillary as isolating environment.Each flushing capillaceous, coating procedure all complete by pressure above, and enter (endpiece from the end of capillary array, one end with respect to injection port), from front end, be discharged to waste liquid pool, the solution tank of described NaOH aqueous solution, polycation aqueous solution, polyanion aqueous solution and citric acid solution is controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
The inlet end capillaceous of capillary array is inserted in array sample groove one to one, 2nL sample is entered to front end capillaceous in capillary array with voltage or pressure mode simultaneously, and the allotment volume ratio of described sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 7.
Kapillary two ends are connected for dissociating buffer system the constant DC voltage that adds 10-30kV on kapillary are as separated driving force, with drive various albumen in sample with friction speed by detection window position.Described array sample groove and dissociating buffer system are controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
Each protein component flowing through in kapillary detects by ultraviolet light array detector, forms and detects collection of illustrative plates, and be recorded in software.
Label using dimethyl sulfoxide as electroosmotic flow, the appearance time at other each peak and dimethyl sulfoxide as a reference, are confirmed the protein component of each peak representative.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out to integration, and the percentage that obtains glycosylated hemoglobin HbAlc is substantially the same manner as Example 1.
Embodiment 5:
Select 10 inwalls to have the elastic quartz capillary tube of polyacrylamide coating (20 microns of internal diameters, long 30cm) and metal electrode to form capillary array.At capillary array rear portion, be provided with transparent detection window, use ultraviolet/visible light to carry out quantitative and qualitative analysis detection to the separated protein component obtaining in kapillary.
The inlet end capillaceous of capillary array is inserted in array sample groove one to one, 50nL sample is entered to front end capillaceous in capillary array with voltage or pressure mode simultaneously, and the allotment volume ratio of described sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 3.
Kapillary two ends are connected for dissociating buffer system the constant DC voltage that adds 10-30kV on kapillary are as separated driving force, with drive various albumen in sample with friction speed by detection window position.Described array sample groove and dissociating buffer system are controlled automatically by the mechanical arm that is connected with control element, also can be by manually controlling.
Each protein component flowing through in kapillary detects by optical arrays detecting device, forms and detects collection of illustrative plates, and be recorded in software.
Label using dimethyl sulfoxide as electroosmotic flow, the appearance time at other each peak and dimethyl sulfoxide as a reference, are confirmed the protein component of each peak representative.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out to integration, and the percentage that obtains glycosylated hemoglobin HbAlc is substantially the same manner as Example 1.
If embodiment 1 is with as shown in embodiment 5, using the capillary array forming without coatings capillary pipe provided by the present invention as split tunnel, after using polyanion coating and polycation coating, its testing result is with directly the cated capillary array of use inwall is basic identical as the testing result of split tunnel.
Claims (17)
1. one kind for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, use array capillary and optical arrays pick-up unit a plurality of samples to be carried out to glycosylated hemoglobin HbAlc detection simultaneously, comprise the following steps:
1) dissociating buffer is filled with in each kapillary in array capillary as isolating environment;
2) each sample is joined simultaneously to each front end capillaceous in array capillary;
3) after sample enters each kapillary front end in capillary array, the two ends of array capillary are replaced by dissociating buffer;
4) at kapillary two ends, apply voltage as separated driving force;
5) by detection light source and optical arrays detecting device, the sample in kapillary is carried out to glycosylated hemoglobin HbAlc detection;
6) according to step 5) gained, detect the content that data calculate the glycosylated hemoglobin HbAlc in each sample;
Before step 1), at the capillary tube inner wall that forms described array capillary, prepare coating, described coating is the duplex coating that polycation coating and polyanion coating form, and the method for preparing described duplex coating is: use said polycation solution and polyanion solution to rinse kapillary; Described said polycation solution is the aqueous solution that contains polycation, is selected from a kind of in polybrene aqueous solution, diallyl dimethyl ammoniumchloride aqueous solution, albumin aqueous solution, and its concentration is 0.001-1wt%; Described polyanion solution is the aqueous solution that contains polyanion, is selected from a kind of in polyacrylic acid aqueous solution, kayexalate aqueous solution, chondroitin sulfate aqueous solution, and its concentration is 0.001-1wt%.
2. as claimed in claim 1 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, capillary inner diameter 20-100 micron in described capillary array, effectively separation length 10-50 centimetre.
3. as claimed in claim 2 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, in described capillary array, quantity capillaceous is 4-48 root, and described kapillary is without coating quartz capillary.
4. as claimed in claim 3 a kind ofly it is characterized in that for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, described each front portion capillaceous is parallel to each other, and end set is to a place.
5. as claimed in claim 1 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, before preparation coating, use the kapillary in strong acid and/or strong base solution pair array kapillary to process.
6. as claimed in claim 5 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, kapillary in described strong acid and/or strong base solution pair array kapillary is processed, polycation and polyanion solution rinse kapillary, dissociating buffer is filled with process capillaceous and all by pressure, completes, described strong acid and strong base solution, polycation and polyanion solution, dissociating buffer all enter from the end of capillary array, from front end, are discharged to waste liquid pool.
7. as claimed in claim 6 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, described strong acid solution is selected from hydrochloride buffer or nitric acid damping fluid, the concentration of described strong acid solution is 0.01-1mol/L, described strong base solution is selected from sodium hydrate aqueous solution or lithium hydroxide aqueous solution, and the concentration of described strong base solution is 0.01-1mol/L.
8. a kind of as described in claim as arbitrary in claim 2-7 is characterized in that for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, and the pH value of described dissociating buffer is 3-5.
9. a kind of as described in claim as arbitrary in claim 2-7 is characterized in that for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, described step 2) in each sample in contain electroendosmotic flow marker.
10. as claimed in claim 9 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, the appearance time of described electroendosmotic flow marker of take is benchmark, draw glycosylated hemoglobin and the corresponding peak of non-glycosylated hemoglobin, the peak value at glycosylated hemoglobin and the corresponding peak of non-glycosylated hemoglobin is carried out obtaining after integration also calculates to the content of glycosylated hemoglobin in sample.
11. is as claimed in claim 10 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, the formula of described calculating is: glycosylated hemoglobin (%) content=glycosylated hemoglobin peak value/(glycosylated hemoglobin peak value+non-glycosylated hemoglobin peak value) * 100%.
12. as claimed in claim 11 a kind ofly it is characterized in that for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, described electroendosmotic flow marker is dimethyl sulfoxide; The volume ratio of described dimethyl sulfoxide and sample is 1:1-30.
13. is as claimed in claim 12 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, described sample comprises blood sample and hemolytic agent, and the volume ratio of described blood sample and hemolytic agent is 1:1-3.
14. kinds as claimed in claim 13 are for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, the method that described each sample joins each front end capillaceous in capillary array is simultaneously voltage system or pressure mode.
A kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously as described in 15. claims as arbitrary in claim 2-7, it is characterized in that, separated driving force in described step 4) is constant dc, and its voltage is 10-30kV.
A kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously as described in 16. claims as arbitrary in claim 2-7, it is characterized in that, the light source in described step 5) is that wavelength is ultraviolet or the visible light source of 180-600nm.
17. is as claimed in claim 16 a kind of for detect the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc simultaneously, it is characterized in that, described ultraviolet or visible light source are Halogen lamp LED or LED light source, and described optical arrays detecting device is the optical arrays detecting device that PMT or CCD detecting device form.
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