CN102539512A - 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 multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc.
Background technology
Detection to glycosylated hemoglobin at present all exists the detection flux low always, the high or low situation of accuracy of cost.
For example the accuracy of immunization is relatively poor, and cost is also than higher; The accuracy of ion-exchange chromatography method is very high, but has no idea to realize the 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 cover high flux, and is high-speed, can detect the capillary array electrophoresis method of a plurality of samples simultaneously, to satisfy the requirement that glycosylated hemoglobin detects.
The multiple-pass capillary tube electrophoresis technology has been successfully applied to dna sequencing system, does not still have report at home but the multiple-pass capillary tube electrophoresis system is used for the detection of glycosylated hemoglobin HbAlc index; Domestic at present, have the report that the single capillary electrophoresis method is used for glycosylated hemoglobin HbAlc detection, but the method flux is low excessively, can't satisfy the requirement that great amount of samples detects.This method combines the multiple-pass capillary tube electrophoresis technology of using in the research work with single track Capillary Electrophoresis glycosylated hemoglobin detection technique; Both solved the difficult problem that glycosylated hemoglobin HbAlc detects; Improve the flux and the speed that detect again greatly, satisfied the requirement of test experience chamber.
Summary of the invention
The present invention is used for glycosylated hemoglobin HbAlc relative content and measures high flux and fast measuring.The present invention uses the array of many capillaries can analyze and detect the glycosylated hemoglobin HbAlc in a plurality of blood samples simultaneously as split tunnel, and automatically the result is analyzed, and generates report.
The present invention provides a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc, uses array capillary and optical arrays pick-up unit simultaneously a plurality of samples to be carried out glycosylated hemoglobin HbAlc and detects.
Preferably, said detection method may further comprise the steps:
1) dissociating buffer is charged in each kapillary in the array capillary as isolating environment;
2) each sample is joined each front end capillaceous in the array capillary simultaneously;
3) after sample gets into each the kapillary front end in the capillary array, dissociating buffer is replaced by at the two ends of array capillary;
4) apply voltage at the kapillary two ends as separating driving force;
5) through detection light source and optical arrays detecting device the sample in the kapillary being carried out glycosylated hemoglobin HbAlc detects;
6) detect the content that data computation draws the glycosylated hemoglobin HbAlc in each sample according to the step 5) gained.
Preferred, capillary inner diameter 20-100 micron in the said capillary array, effectively separation length 10-50 centimetre.
Preferred, quantity capillaceous is the 4-48 root in the said capillary array, and said kapillary is no coating quartz capillary or the cated neutral quartz capillary of inwall.
Preferred, internal diameter and length that said kapillary is every all are consistent.
Further, said 4-48 capillary constitutes capillary array as split tunnel, can carry out electrophoretic separation and detection to 4-48 sample while sample introduction.
Preferred, said each front portion capillaceous is parallel to each other, and end is gathered a place.
Preferred, before step 1), prepare coating at the capillary tube inner wall that constitutes said array capillary.
Preferred, said coating is the duplex coating that polycation coating and polyanion coating are constituted.
Preferred, the method for preparing said duplex coating is: wash kapillary with said polycation solution earlier, again with polyanion solution flushing kapillary.
Preferred, before the preparation coating, use the kapillary in strong acid and/or the strong base solution array kapillary to handle.
Preferred; Kapillary in said strong acid and/or the strong base solution array kapillary is handled, polycation and polyanion solution flushing kapillary, dissociating buffer charge into process capillaceous and all accomplish through pressure; Said strong acid and strong base solution, polycation and polyanion solution, dissociating buffer all get into from the end of capillary array, are discharged to waste liquid pool from front end.
Preferred, said strong acid solution is selected from hydrochloride buffer or nitric acid damping fluid, and the concentration of said strong acid solution is 0.01-1mol/L.
Preferred, said strong base solution is selected from sodium hydrate aqueous solution or lithium hydroxide aqueous solution, and the concentration of said strong base solution is 0.01-1mol/L.
Preferred, said said polycation solution is the WS that contains polycation, is selected from the polybrene WS, the diallyl dimethyl ammoniumchloride WS, the albumin WS, and its concentration is 0.001-1wt%.
Preferred, said polyanion solution is the WS that contains polyanion, is selected from the polyacrylic acid WS, the kayexalate WS, the chondroitin sulfate WS, and its concentration is 0.001-1wt%.
Because uncoated kapillary can be adsorbed onto inside surface capillaceous with sample, can not isolate the peak of glycosylated hemoglobin HbAlc usually, thereby can not go out the peak.So the present invention uses polyanion coating and polycation coating to adsorb at capillary tube inner wall with the albumen that reaches in the control blood sample on no coatings capillary pipe tube wall, and accelerates the purpose that peak speed of glycosylated hemoglobin.Multiple-pass capillary tube electrophoresis system involved in the present invention uses the capillary array of no coatings capillary pipe composition as split tunnel, to reach the reduction testing cost, keeps the purpose than the long life capillaceous simultaneously.But multiple-pass capillary tube electrophoresis system involved in the present invention also can directly use the cated capillary array of inwall as split tunnel (for example can select the kapillary of coatings such as polyacrylamide, polyvinyl alcohol (PVA), polypyrrole alkane ketone for use), also can on the cated kapillary of inwall, re-use polyanion coating or polycation coating.
Preferred, the pH value of said dissociating buffer is 3-5.
Preferred, said step 2) contains electroendosmotic flow marker in each sample in.
Preferred, be benchmark with the appearance time of said electroendosmotic flow marker, draw glycosylated hemoglobin and the pairing peak of non-glycosylated hemoglobin, peak value is carried out the content that integration obtains glycosylated hemoglobin in the sample.
Preferred; Appearance time with said electroendosmotic flow marker is a benchmark; Draw glycosylated hemoglobin and the pairing peak of non-glycosylated hemoglobin, the peak value at glycosylated hemoglobin and the pairing peak of non-glycosylated hemoglobin is carried out obtaining after integration also calculates the content of glycosylated hemoglobin in the sample.
Preferred, the formula of said calculating is: glycosylated hemoglobin (mol%) content=glycosylated hemoglobin peak value/(glycosylated hemoglobin peak value+non-glycosylated hemoglobin peak value) * 100%.
Preferred, electroendosmotic flow marker is a dimethyl sulfoxide in the said step 3); The volume ratio of said dimethyl sulfoxide and sample is 1: 1-30.
Preferred, the sample in the said step 3) comprises blood sample and hemolytic agent, and the volume ratio of said blood sample and hemolytic agent is 1: 1-3, said hemolytic agent can be commercially available commercialization hemolytic agent.
Preferred, to join the method for each front end capillaceous in the capillary array simultaneously be voltage system or pressure mode to each sample in the said step 3).
Preferred, the sample size of said sample is 1-100nL.
Preferred, the mode that applies voltage in the said step 4) is in the dissociating buffer that electrode is inserted the kapillary two ends or directly be connected on the kapillary two ends.
Preferred, the voltage that is applied in the said step 4) is constant dc, and U=10-30kV.
Preferred, the light source in the said step 5) is that wavelength is ultraviolet or the visible light source of 180-600nm.
Preferred; Said ultraviolet or visible light source are Halogen lamp LED or led light source; Said optical arrays detecting device comprises a plurality of photodetectors, and said photodetector and kapillary mate one by one, and said optical arrays detecting device is the optical arrays detecting device that PMT or CCD detecting device are formed.
Choosing is more arranged, and said peak value is the uv absorption peak value.
Because the two ends of capillary array have applied High Level DC Voltage (10-30kV); Under the High Level DC Voltage effect; Blood protein in every capillary can begin electrophoresis under the electric field force effect, but because electrophoretic velocity difference, glycosylated hemoglobin HbAlc can separate with other kinds proteinoid.The afterbody of parallel-segment capillaceous is provided with transparent detection window in the said capillary array, and said effective separation length is that the capillary inlet is to the distance between the detection window.Because each effective separation length capillaceous all equates, so the deposition condition of each sample is basic identical, it is basic identical to that is to say that same composition in each sample arrives time of transparent detection window.The light signal that detection light source is sent is perpendicular to detection window; And through the detection window arrival photodetector corresponding with this detection window; When electroendosmotic flow marker, glycosylated hemoglobin HbAlc and other kinds albuminoid pass through transparent detection window; Photodetector in the optical arrays detecting device corresponding with this detection window can be noted the response of each component and will detect data and be sent to signal analysis unit, and signal analysis unit draws the absorption peak peak value of each component after with the gained data processing.
Description of drawings
Fig. 1 is principle of the invention synoptic diagram (is example with the single capillary)
Fig. 2 is testing result synoptic diagram of the present invention (is example with the single capillary)
Embodiment
Multiple-pass capillary tube electrophoresis system involved in the present invention can use the capillary array of no coatings capillary pipe composition as split tunnel, to reach the reduction testing cost, keeps the purpose than the long life capillaceous simultaneously.Said capillary array comprises 4-48 root elastic quartz capillary tube (no coating, internal diameter 20-100 micron, length 10-50cm), and each front portion capillaceous is parallel to each other, and end is gathered a place.The present invention also simultaneously forms polycation and polyanion coating at capillary tube inner wall, goes out peak speed with the quickening glycosylated hemoglobin.Each components contents detects through ultraviolet optical arrays detecting device in the said sample.
The principle of the principle of the invention (is example with the single capillary) as shown in Figure 1.Principle of the invention synoptic diagram as shown in Figure 1 comprises sample injection unit, optical detection unit and signal analysis unit 8, and said sample injection unit matches with said optical detection unit, and said optical detection unit is electrically connected with signal analysis unit 8.
Further, said optical detection unit comprises detection light source 7, optical arrays detecting device 3 and array capillary bundle, and said array capillary bundle matches with said sample injection unit, and detection light source 7 matches with said array capillary bundle with optical arrays detecting device 3.
Further, the inwall that constitutes the kapillary 9 of said array capillary bundle is provided with polycation coating and polyanion coating.
Further, said array capillary bundle is made up of 4-48 capillary 9.
Further, be parallel to each other between the kapillary 9 of said forming array capillary bundle, an end of kapillary 9 is that front end 1, the other end are terminal 2; And terminal 2 are collected to a place.
Further, the kapillary 9 of said 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 said forming array capillary bundle is an internal diameter 20-100 micron.
Further set forth the present invention below in conjunction with specific embodiment, should be understood that these embodiment only are used to the present invention is described and are not used in restriction protection scope of the present invention.
Embodiment 1:
Select for use 10 elastic quartz capillary tubes (20 microns of internal diameters, long 30cm) and metal electrode to constitute capillary array.Be provided with transparent detection window at the capillary array rear portion, use ultraviolet light to carry out qualitative and detection by quantitative separating the protein component that obtains in the kapillary.
After the elastic quartz capillary tube 0.1mol/L NaOH WS is handled, more successively with polycation (diallyl dimethyl ammoniumchloride, 0.5wt%) and polyanion (chondroitin sulfate, 0.5wt%) WS washes kapillary, obtains duplex coating.Citric acid solution with pH=4 charges into kapillary as isolating environment subsequently.More than each flushing capillaceous, coating procedure all accomplish through pressure; And get into (endpiece from the end of capillary array; End with respect to injection port); Be discharged to waste liquid pool from front end, the solution tank of the said NaOH WS, the polycation WS, the polyanion WS and citric acid solution is controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
The inlet end capillaceous of capillary array is inserted in the array sample groove one to one; The 50nL sample is got into front end capillaceous in the capillary array simultaneously with voltage or pressure mode, and the allotment volume ratio of said sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 3.
With the kapillary two ends connect for separation buffer liquid systems and the constant DC voltage that on kapillary, adds 10-30kV as separating driving force, with drive various albumen in the sample with friction speed through the detection window position.Said array sample groove and separation buffer liquid systems are controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
Each protein component that flows through in the kapillary detects through the ultraviolet light array detector, forms to detect collection of illustrative plates, and is recorded in the software.
With the label of dimethyl sulfoxide as EOF, the appearance time at other each peak and dimethyl sulfoxide are confirmed the protein component of each peak representative as a reference, its testing result (is example with the single capillary) as shown in Figure 2.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out integration, obtain the percentage of glycosylated hemoglobin HbAlc.
Embodiment 2:
Select for use 4 elastic quartz capillary tubes (50 microns of internal diameters, long 50cm) and metal electrode to constitute capillary array.Be provided with transparent detection window at the capillary array rear portion, use ultraviolet light to carry out qualitative and detection by quantitative separating the protein component that obtains in the kapillary.
After the elastic quartz capillary tube 1mol/L KOH WS is handled, more successively with polycation (the polybrene WS, 1wt%) and polyanion (the polyacrylic acid WS, 0.001wt%) WS washes kapillary, obtains duplex coating.Citric acid solution with pH=5 charges into kapillary as isolating environment subsequently.More than each flushing capillaceous, coating procedure all accomplish through pressure; And get into (endpiece from the end of capillary array; End with respect to injection port); Be discharged to waste liquid pool from front end, the solution tank of the said NaOH WS, the polycation WS, the polyanion WS and citric acid solution is controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
The inlet end capillaceous of capillary array is inserted in the array sample groove one to one; The 100nL sample is got into front end capillaceous in the capillary array simultaneously with voltage or pressure mode, and the allotment volume ratio of said sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 9: 21.
With the kapillary two ends connect for separation buffer liquid systems and the constant DC voltage that on kapillary, adds 10-30kV as separating driving force, with drive various albumen in the sample with friction speed through the detection window position.Said array sample groove and separation buffer liquid systems are controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
Each protein component that flows through in the kapillary detects through the ultraviolet light array detector, forms to detect collection of illustrative plates, and is recorded in the software.
With the label of dimethyl sulfoxide as EOF, the appearance time at other each peak and dimethyl sulfoxide are confirmed the protein component of each peak representative as a reference.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out integration, and the percentage and the embodiment 1 that obtain glycosylated hemoglobin HbAlc are basic identical.
Embodiment 3:
Select for use 30 elastic quartz capillary tubes (75 microns of internal diameters, long 10cm) and metal electrode to constitute capillary array.Be provided with transparent detection window at the capillary array rear portion, use visible light to carry out qualitative and detection by quantitative separating the protein component that obtains in the kapillary.
After the elastic quartz capillary tube 0.1mol/L KOH WS is handled, more successively with polycation (the albumin WS, 0.1wt%) and polyanion (chondroitin sulfate, 0.05wt%) WS washes kapillary, obtains duplex coating.Citric acid solution with pH=3 charges into kapillary as isolating environment subsequently.More than each flushing capillaceous, coating procedure all accomplish through pressure; And get into (endpiece from the end of capillary array; End with respect to injection port); Be discharged to waste liquid pool from front end, the solution tank of the said NaOH WS, the polycation WS, the polyanion WS and citric acid solution is controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
The inlet end capillaceous of capillary array is inserted in the array sample groove one to one; The 10nL sample is got into front end capillaceous in the capillary array simultaneously with voltage or pressure mode, and the allotment volume ratio of said sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 10: 10.
With the kapillary two ends connect for separation buffer liquid systems and the constant DC voltage that on kapillary, adds 10-30kV as separating driving force, with drive various albumen in the sample with friction speed through the detection window position.Said array sample groove and separation buffer liquid systems are controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
Each protein component that flows through in the kapillary detects through visible light optical arrays detecting device, forms to detect collection of illustrative plates, and is recorded in the software.
With the label of dimethyl sulfoxide as EOF, the appearance time at other each peak and dimethyl sulfoxide are confirmed the protein component of each peak representative as a reference.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out integration, and the percentage and the embodiment 1 that obtain glycosylated hemoglobin HbAlc are basic identical.
Embodiment 4:
Select for use 48 elastic quartz capillary tubes (100 microns of internal diameters, long 40cm) and metal electrode to constitute capillary array.Be provided with transparent detection window at the capillary array rear portion, use ultraviolet light to carry out qualitative and detection by quantitative separating the protein component that obtains in the kapillary.
After the elastic quartz capillary tube 0.01mol/L NaOH WS is handled; Again successively with polycation (diallyl dimethyl ammoniumchloride; 0.001wt%) and polyanion (the kayexalate WS, 1wt%) WS flushing kapillary obtains duplex coating.Citric acid solution with pH=5 charges into kapillary as isolating environment subsequently.More than each flushing capillaceous, coating procedure all accomplish through pressure; And get into (endpiece from the end of capillary array; End with respect to injection port); Be discharged to waste liquid pool from front end, the solution tank of the said NaOH WS, the polycation WS, the polyanion WS and citric acid solution is controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
The inlet end capillaceous of capillary array is inserted in the array sample groove one to one; The 2nL sample is got into front end capillaceous in the capillary array simultaneously with voltage or pressure mode, and the allotment volume ratio of said sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 7.
With the kapillary two ends connect for separation buffer liquid systems and the constant DC voltage that on kapillary, adds 10-30kV as separating driving force, with drive various albumen in the sample with friction speed through the detection window position.Said array sample groove and separation buffer liquid systems are controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
Each protein component that flows through in the kapillary detects through the ultraviolet light array detector, forms to detect collection of illustrative plates, and is recorded in the software.
With the label of dimethyl sulfoxide as EOF, the appearance time at other each peak and dimethyl sulfoxide are confirmed the protein component of each peak representative as a reference.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out integration, and the percentage and the embodiment 1 that obtain glycosylated hemoglobin HbAlc are basic identical.
Embodiment 5:
Select for use 10 inwalls to have elastic quartz capillary tube of polyacrylamide coating (20 microns of internal diameters, long 30cm) and metal electrode to constitute capillary array.Be provided with transparent detection window at the capillary array rear portion, use ultraviolet to carry out qualitative and detection by quantitative separating the protein component that obtains in the kapillary.
The inlet end capillaceous of capillary array is inserted in the array sample groove one to one; The 50nL sample is got into front end capillaceous in the capillary array simultaneously with voltage or pressure mode, and the allotment volume ratio of said sample is: blood sample: hemolytic agent: dimethyl sulfoxide=1: 3: 3.
With the kapillary two ends connect for separation buffer liquid systems and the constant DC voltage that on kapillary, adds 10-30kV as separating driving force, with drive various albumen in the sample with friction speed through the detection window position.Said array sample groove and separation buffer liquid systems are controlled by the mechanical arm that is connected with control element automatically, also can be through manually control.
Each protein component that flows through in the kapillary detects through the optical arrays detecting device, forms to detect collection of illustrative plates, and is recorded in the software.
With the label of dimethyl sulfoxide as EOF, the appearance time at other each peak and dimethyl sulfoxide are confirmed the protein component of each peak representative as a reference.The peak value of glycosylated hemoglobin HbAlc and non-glycosylated hemoglobin HbA0 is carried out integration, and the percentage and the embodiment 1 that obtain glycosylated hemoglobin HbAlc are basic identical.
Shown in embodiment 1 and embodiment 5; The capillary array that no coatings capillary pipe provided by the present invention is formed is 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 (23)
1. a multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc is characterized in that, uses array capillary and optical arrays pick-up unit simultaneously a plurality of samples to be carried out glycosylated hemoglobin HbAlc and detects.
2. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 1 may further comprise the steps:
A) dissociating buffer is charged in each kapillary in the array capillary as isolating environment;
B) each sample is joined each front end capillaceous in the array capillary simultaneously;
C) after sample gets into each the kapillary front end in the capillary array, dissociating buffer is replaced by at the two ends of array capillary;
D) apply voltage at the kapillary two ends as separating driving force;
E) through detection light source and optical arrays detecting device the sample in the kapillary being carried out glycosylated hemoglobin HbAlc detects;
F) detect the content that data computation draws the glycosylated hemoglobin HbAlc in each sample according to the step 5) gained.
3. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 2; It is characterized in that; Capillary inner diameter 20-100 micron in the said capillary array, effectively separation length 10-50 centimetre.
4. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 3; It is characterized in that; Quantity capillaceous is the 4-48 root in the said capillary array, and said kapillary is no coating quartz capillary or the cated neutral quartz capillary of inwall.
5. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 4 is characterized in that said each front portion capillaceous is parallel to each other, and end is gathered a place.
6. like the described a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc of claim 2-5, it is characterized in that, before step 1), prepare coating at the capillary tube inner wall that constitutes said array capillary.
7. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 6 is characterized in that said coating is the duplex coating that polycation coating and polyanion coating are constituted.
8. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 7; It is characterized in that the method for preparing said duplex coating is: use said polycation solution and polyanion solution flushing kapillary.
9. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 8; It is characterized in that; Said said polycation solution is the WS that contains polycation; Be selected from the polybrene WS, the diallyl dimethyl ammoniumchloride WS, the albumin WS, its concentration is 0.001-1wt%.
10. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 8; It is characterized in that; Said polyanion solution is the WS that contains polyanion; Be selected from the polyacrylic acid WS, the kayexalate WS, the chondroitin sulfate WS, its concentration is 0.001-1wt%.
11. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 8; It is characterized in that; Before the preparation coating, use the kapillary in strong acid and/or the strong base solution array kapillary to handle.
12. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 11; It is characterized in that; Kapillary in said strong acid and/or the strong base solution array kapillary is handled, polycation and polyanion solution flushing kapillary, dissociating buffer charge into process capillaceous and all accomplish through pressure; Said strong acid and strong base solution, polycation and polyanion solution, dissociating buffer all get into from the end of capillary array, are discharged to waste liquid pool from front end.
13. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 12; It is characterized in that; Said strong acid solution is selected from hydrochloride buffer or nitric acid damping fluid; The concentration of said strong acid solution is 0.01-1mol/L, and said strong base solution is selected from sodium hydrate aqueous solution or lithium hydroxide aqueous solution, and the concentration of said strong base solution is 0.01-1mol/L.
14., it is characterized in that the pH value of said dissociating buffer is 3-5 like claim 2-5 or the described a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc of the arbitrary claim of 7-13.
15. like claim 2-5 or the described a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc of the arbitrary claim of 7-13; It is characterized in that said step 2) in each sample in contain electroendosmotic flow marker.
16. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 15; It is characterized in that; Appearance time with said electroendosmotic flow marker is a benchmark; Draw glycosylated hemoglobin and the pairing peak of non-glycosylated hemoglobin, the peak value at glycosylated hemoglobin and the pairing peak of non-glycosylated hemoglobin is carried out obtaining after integration also calculates the content of glycosylated hemoglobin in the sample.
17. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 16; It is characterized in that the formula of said calculating is: glycosylated hemoglobin (%) content=glycosylated hemoglobin peak value/(glycosylated hemoglobin peak value+non-glycosylated hemoglobin peak value) * 100%.
18. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 17 is characterized in that said electroendosmotic flow marker is a dimethyl sulfoxide; The volume ratio of said dimethyl sulfoxide and sample is 1: 1-30.
19. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 18; It is characterized in that; Said sample comprises blood sample and hemolytic agent, and the volume ratio of said blood sample and hemolytic agent is 1: 1-3.
20. kind as claimed in claim 19 is used for detecting simultaneously the multiple-pass capillary tube electrophoresis detection method of a plurality of sample glycosylated hemoglobin HbAlc; It is characterized in that the method that said each sample joins each front end capillaceous in the capillary array simultaneously is voltage system or pressure mode.
21. like claim 2-5 or the described a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc of the arbitrary claim of 7-13; It is characterized in that; Separation driving force in the said step 4) is a constant dc, and its voltage is 10-30kV.
22. like claim 2-5 or the described a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc of the arbitrary claim of 7-13; It is characterized in that the light source in the said step 5) is that wavelength is ultraviolet or the visible light source of 180-600nm.
23. a kind of multiple-pass capillary tube electrophoresis detection method that is used for detecting simultaneously a plurality of sample glycosylated hemoglobin HbAlc as claimed in claim 22; It is characterized in that; Said ultraviolet or visible light source are Halogen lamp LED or led light source, and said optical arrays detecting device is the optical arrays detecting device that PMT or CCD detecting device are formed.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105806919A (en) * | 2015-01-15 | 2016-07-27 | 爱科来株式会社 | Hemoglobin analysis method and solution therefor |
| CN107944221A (en) * | 2017-11-21 | 2018-04-20 | 南京溯远基因科技有限公司 | A kind of stitching algorithm of parallel seperated nuclear acid segment and its application |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105806919A (en) * | 2015-01-15 | 2016-07-27 | 爱科来株式会社 | Hemoglobin analysis method and solution therefor |
| CN105806919B (en) * | 2015-01-15 | 2019-07-05 | 爱科来株式会社 | The analysis method of sample and its used in solution |
| CN107944221A (en) * | 2017-11-21 | 2018-04-20 | 南京溯远基因科技有限公司 | A kind of stitching algorithm of parallel seperated nuclear acid segment and its application |
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