Method and kit for detecting antiasthmatic drugs in serum
A technology for anti-asthma and drugs, which is applied in the field of detection of anti-asthma drugs in serum, can solve the problems of good linearity, high feasibility, low detection signal, and low sensitivity, and achieve the goal of reducing time detection cost, less reagent consumption, and good extraction effect Effect
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Embodiment 1
[0058] 1. Extract theophylline, dyphylline and doxofylline in serum
[0059] (1) Add 25 μL serum sample to the centrifuge tube, add 100 μL extract solution, the extract solution is a mixture of methanol, acetonitrile and trichloroacetic acid solution, the volume ratio of methanol and acetonitrile is 1:5 (the volume fraction of trichloroacetic acid is 0.05 %).
[0060] (2) The above-mentioned sample solution was vortexed for 5 minutes, and the speed of the centrifuge was 12000 r / min; centrifuged for 5 minutes.
[0061] (3) After centrifugation, take 10 μL of supernatant to a 96-well receiving plate, add 100 μL of diluent, and mix well. The composition of the diluent is an aqueous solution containing 0.05% trichloroacetic acid by volume.
[0062] (4) Cover with an aluminum foil sealing film for chromatographic analysis.
[0063] 2. Instrument method
[0064] (1) Chromatographic conditions: chromatographic column: ZORBAX SB C18 (2.1mm×100mm, 3.5μm), column temperature: 40°C; m...
Embodiment 2
[0100] Embodiment 2 standard curve linear analysis
[0101] Add mixed standard to bovine serum, the concentration of theophylline, diprophylline and doxofylline are 20 μg / mL, 10 μg / mL, 10 μg / mL in sequence. Using bovine serum as the diluent, the bovine serum standard mixture was sequentially diluted 1:1 in equal proportions, with a total of 6 concentration gradients, and the method in Example 1 was used for detection.
[0102] Use data processing software to data processing, calculate and draw standard curve, wherein, the standard curve figure of theophylline, diprophylline, doxofylline is as follows Figure 13 , Figure 14 and Figure 15 As shown, the correlation coefficient R2 is greater than 0.999, which meets the linearity requirement.
[0103] Among them, the linear range of theophylline is 0.1-80 μg / mL, and the linear range of diprophylline and doxofylline is 0.05-40 μg / mL.
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