Concentration determination of diazepam in human serum by HPLC and clinical application
Objective To establish a HPLC method for determination of diazepam concentration in human serum and determination of the plasma concentration in clinical patients after intramuscular injection of diazepam.Methods Human serum samples were extracted by 5 mL n-pentane and analyzed.Column:SB-C18 column (150 mm×4.6 mm,5 μm);Mobile phase:methanol-water-tetramethylethylenediamine-acetic acid (670:330:2.2:1.76);The column temperature was 40 ℃;Flow rate:0.8 mL·min-1;and the detection wavelength was 254 nm;Injection volume was 20 μL.Clozapine was used as internal standard.Results The calibration curves were linear in the range of 10-1 200 μg·L-1 for diazepam(r=0.998 47).The lower limit of quantification was 10 μg·L-1.The average recoveries of diazepam at high,middle and low concentration over 95%,the extraction recovery over 70%.The intra-day and inter-day RSD were less than 15% (n=5).The diazepam stability of RSD were less than 15% (n=5).The calibration curve of good linearity,r=0.998 47.The regression equation of paroxetine was Y=2 205.2X+1 876.1.After 20 patients were clinical intramuscular injection of diazepam 20 mg bid,before five days the morning treatment for determination of diazepam concentration in human serum.ConclusionsThe method is sensitive,simple,rapid,accurate and can be used for determination of diazepam concentration in human serum and can provide reference for clinical patient individualized.
NETL
NSTL
万方数据
维普
