The detection method of clonazepam
A clonazepam and detection method technology, which is applied in the detection field of clonazepam, can solve the problems of time-consuming and long sample detection time, etc.
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Embodiment 1
[0095] Example 1: Preparation of standard solutions of series concentrations
[0096] (a) Preparation of standard stock solution:
[0097] Accurately weigh 5 mg of standard clonazepam into a 5 mL volumetric flask, dissolve with methanol: water = 7: 3 diluent, and dilute to 5 mL to obtain a standard stock solution, which is stored at -80 °C, Valid for one year.
[0098] (b) Preparation of standard working solution
[0099] Take an appropriate amount of standard stock solution A in step (a), dilute and mix with a diluent of methanol: water = 7:3 to obtain a series of standard mixed intermediate solution C containing 50-2000ng / mL clonazepam, and in - Stored at 80°C, the validity period is 3 months;
[0100] Among them, the standard working solutions of different concentrations are a series of solutions containing clonazepam: 50ng / mL, 100ng / mL, 200ng / mL, 400ng / mL, 800ng / mL, 1000ng / mL, and 2000ng / mL.
[0101] (c) Preparation of internal standard stock solution
[0102] Take 5m...
Embodiment 2
[0107] Example 2: Fitting the Standard Curve Equation
[0108] Seven kinds of standard solutions in Example 1 were detected respectively by liquid chromatography-mass spectrometry, and the chromatograms of the standard solutions of seven kinds of clonazepam with different concentrations were obtained.
[0109] From the chromatograms of the standard solutions of the above-mentioned clonazepam, the peak areas corresponding to clonazepam and the internal standard in the seven standard solutions were obtained respectively. The ratio of the peak area of pan to the chromatographic peak area of the internal standard is used as the ordinate y1 of the standard curve equation, and the abscissa x1 of the standard curve equation with the concentration in the above-mentioned clonazepam standard working solution and the concentration of the internal standard, Perform linear regression on the data of different concentrations obtained from the above detection, and the standard curve equat...
Embodiment 3
[0123] Example 3: Treatment of samples to be tested
[0124] 3.1 Take at least 2ml of blood to be treated, centrifuge at 3500rpm for 10min, take the supernatant to obtain serum or plasma, and store the above-mentioned serum or plasma at -20°C for use before analysis.
[0125] 3.2 Use a pipette to pipette 10 μL of the internal standard in Example 1 into a 1.5 mL centrifuge tube, then add 100 μL of the serum or plasma from step 3.1, vortex and mix for 0.5 min at 2000 rpm, then add 800 μL of ethyl acetate was mixed by vortexing at 2000 rpm for 5 min, then centrifuged at high speed for 10 min at 12000 rpm, and 600 μL of the centrifuged supernatant was transferred to a new 1.5 mL centrifuge tube. The 1.5 mL centrifuge tube was transferred to a nitrogen drying device, the supernatant was dried, 100 μL of methanol was transferred to the 1.5 mL centrifuge tube where the supernatant was dried, and the obtained supernatant was vortexed at 2000 rpm for 2 min. The clear liquid is the sam...
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