Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof
A chemiluminescence detection and acridinium ester marker technology, which is applied in the field of life science and medical detection, can solve the problems that the instrument has not been widely used, the analysis speed is slow, and the operation is cumbersome, so as to achieve high detection sensitivity, simple equipment, The effect of extending the range of application
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Embodiment 1
[0027] 1. Detection of 4-(2-hydroxysuccinamidocarbonylethyl)phenyl-10-methylacridine-9-carboxylate fluorosulfonate (AE-NHS):
[0028] 1) Electrophoresis conditions: the total length of the separation capillary is 550 mm, the inner diameter is 75 μm; the running buffer is 5 mmol / L, phosphate buffer solution with pH=8.0; the new capillary is washed with 1 mol / L HCl, 1 mol / L NaOH and water in sequence Activate the surface of the column for 20 minutes, wash with 0.1mol / L NaOH and water for 10 minutes before starting work every day, and then wash with running buffer for 5 minutes. Operating voltage: 15kV. Separation temperature: 20°C. Sampling: electric injection 20s.
[0029] 2) Sample detection: use the electrophoresis buffer described in 1) to prepare concentrations of 1.0×10 -11 mol / L, 1.0×10 -10 mol / L, 1.0×10 -9 mol / L, 1.0×10 -8 mol / L, 1.0×10 -7mol / L, 1.0×10 -6 mol / L, each 1ml of AE-NHS standard solution, according to the electrophoresis conditions described in step 1)...
Embodiment 2
[0031] Qualitative and quantitative determination of alanine and lysine:
[0032] 1) Electrophoresis conditions: the total length of the separation capillary is 550mm, and the inner diameter is 75 μm; the running buffer is 20mmol / L, and the acetate buffer solution of pH=5.8 (wherein adding acetonitrile with a volume fraction of 28% is used as an additive); the new capillary uses 1mol / L Rinse the surface of the activation column for 20 min with L of HCl, 1 mol / L NaOH and water in sequence, and rinse with 0.1 mol / L NaOH and water for 10 min before starting work every day, and then rinse with running buffer for 5 min. Operating voltage: 15kV. Separation temperature: 20°C. Sampling: electric injection 15s.
[0033] 2) Luminescent labeling of alanine and lysine standards:
[0034] Weigh 4.0 nmol of alanine and lysine and 2 μl of 2.0 mmol / L AE-NHS in 98 μl of 0.05 mol / L phosphate buffer solution (pH=8.0) for 10-20 minutes at room temperature in the dark.
[0035] 3) Qualitative ...
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