ABSTRACT
OBJECTIVE@#To propose a method for simultaneous determination of codeine(COD), 6-monoacetyl-morphine (6-MAM), morphine (MOR), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in human urine by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS).@*METHODS@#After precipitation of protein by acetonitrile, the urine samples, with added the morphine-d3 (MOR-d3) and morphine-3-Glucuronide-d3 (M3G-d3) as internal standards, were pre-treated by Sirocco protein precipitation plate, and then analyzed by UPLC-MS/MS.@*RESULTS@#The limit of detection was 0.2 ng/mL for both COD and MAM, the limit of quantitation was 0.5 ng/mL for both COD and MAM. The limit of detection was 0.5 ng/mL for MOR, M3G and M6G, the limit of quantitation was 1 ng/mL for them. The linear correlation coefficients were not less than 0.9997, both the inter-day and intra-day precisions were less than 10%, the recoveries were in the range of 70.0% to 98.3%, the matrix effects were about 50.5% to 99.0%.@*CONCLUSION@#This proposed method is simple, rapid and accurate, it could be applied in forensic toxicological analysis.
Subject(s)
Humans , Chromatography, Liquid/methods , Codeine/urine , Limit of Detection , Morphine/urine , Morphine Derivatives/urine , Reproducibility of Results , Sensitivity and Specificity , Substance Abuse Detection/methods , Tandem Mass Spectrometry/methodsABSTRACT
OBJECTIVE@#To develop a column-switching high-performance liquid chromatographic method for the determination of morphine and O6-monoacetylmorphine in urine.@*METHODS@#Urine samples (1.0 ml) were spiked with 1.0 ml borate buffer, after centrifugation, 1.0 ml of supernate were injected directly into an extraction column (YWG C18 33 mm x 5.0 mm, 10 microns). After a washing step with the extraction mobile phase, the retained morphine and O6-monoacetylmorphine were flushed into the analytical column (Lichrospher 100 CN 125 mm x 4.0 mm, 5 microns) with the mobile phase CH3OH-H2O (60:40). The analytical mobile phase is CH3OH-phosphate buffer (pH6.86) (22:78). The UV detector was set at lambda 286 nm.@*RESULTS@#The method shows excellent linearity from 50 to 1,600 ng/ml for morphine and from 100 to 1,600 ng/ml for O6-monoacetylmorphine. The linear correlation coefficients were > 0.999. The relative standard deviations were < 4%. The limits of detection were 40 ng/ml for both morphine and O6-monoacetylmorphine.@*CONCLUSION@#The method described is sensitive, rapid, reproducible, and simple.