| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
In the present study, we established a procedure for the determination of psychotropic drugs, phenothiazines, butyrophenones and benzodiazepines, in human samples by in-tube solid-phase microextraction (SPME)/high-performance liquid chromatography (LC)/mass spectrometry (MS).
In-tube SPME was achieved with SupelcoWaxl0 capillary column (60cm, 0.32mm id., 0.25μm film thickness) for phenothiazines and butyrophenones, and Supelco-Qplot capillary capillary column (50cm, 0.32mm id, 0.25 μm film thickness) for benzodiazepines. The optimum extraction conditions for phenothiazines and butyrophenones from serum and urine samples were 15-20 draw/eject cycles of 30-35 μl of the sample containing Tris-HC1 (pH 8) at a flow rate of 0.1 ml/min. The chromatographic separation of phenothiazines and butyrophenones was carried out with a Capcell Pak C_<18> column (150mm, 2mm i.d.) with 10mM ammonium formate/0.1% formate/acetonitrile as a mobile phase at a flow rate of 0.2ml/min. For benzodiazepines, the o
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ptimum extraction conditions from serum and urine samples were 35 draw/eject cycles of 35 μl of the sample containing Tris-HC1 (pH 8.5) at a flow rate of 0.1 ml/min. The chromatographic separation of benzodiazepines was carried out with a Symmetry Shield RP_<18> column (150mm, 2.1mm i.d.) with 50mM ammonium formate/0.1% formic acid/methanol as a mobile phase at a flow rate of 0.2ml/min. LC/MS with electrospray ionization as interface and positive mode were used for detection of all drugs. In the present method, LC/tandem MS (MS/MS) showed much higher sensitivity than LC/MS. The detection limits of all drugs were 20ng-2 μg/ml for serum and 0.5-1 ng/ml for urine using LC/MS/MS with selected reaction monitoring (SRM). In addition to the spiked human body fluids, the method was applied to serum sample after oral administration of phenothiazine (perazine) by a male volunteer; the drug could be isolated from serum sample by in-tube SPME and sensitively quantified by SRM.
The present in-tube SPME/LC/MS method can be recommended for use in therapeutic monitoring, clinical toxicology and forensic toxicology. Less
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