井上 博之

A simple and low-cost chemical synthetic method for p-hydroxymethamphetamine glucuronide using Lewis acid catalyst has been developed. The formation of by-product was minimized by using methyl 1,2,3,4-tetra-O-acetyl-β-D-glucuronate and boron trifluoride as glucuronyl donor and Lewis acid catalyst, respectively. In this combination, the yield of desired product was improved with the usage of three equivalent of glucuronate donor. The yield of p-hydroxymethamphetamine glucuronide in an optimized condition was about 50%, and was comparable to that of the previous methods.

Simultaneous analytical methods for 18 compounds of fentanyl and its analogues by thin-layer chromatography (TLC), gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) were developed. In TLC, fentanyl analogues were well separated by using toluene-acetone-28% aq. ammonia (20:10:0.3, by vol.) as a developing solvent. In GC/MS, fentanyl analogues, except for fentanyl and acetyl-α-methylfentanyl, could be separated on the extracted ion chromatograms (EIC) of the characteristic fragment ions of each compound. In LC/MS, fentanyl analogues could be separated on the EICs of the protonated molecule of each compound. All of the fentanyl analogues tested were identified correctly by using the combination of TLC, GC/MS and LC/MS.

© 2015 Elsevier Ireland Ltd. Chiral capillary electrophoresis/tandem mass spectrometry (CE/MS/MS) using a chemically modified capillary containing sulfonated groups was developed for the following 8 amphetamine-type stimulants (ATS): amphetamine, methamphetamine (MA), norephedrine, norpseudoephedrine, ephedrine (EP), pseudoephedrine (pEP), dimethylamphetamine and methylephedrine. The running buffer was 10. mM formic acid containing 20. mM highly sulfated γ-cyclodextrin (pH 2.5) as the chiral selector. All 16 enantiomers were well resolved within 60. min, and precisely identified due to their characteristic mass spectra. Further, the RSDs of the migration times of the analytes were no more than 0.3% without any standardization. (1. R,2. S)-(-)-EP and (1. S,2. S)-(+)-pEP, which are important ATS impurities originating in the precursors, were added to a highly concentrated MA solution (1. mg/mL) and analyzed as mock samples for MA impurity analysis. Acceptable repeatability of the migration times of (-)-EP and (+)-pEP (ca. 0.3% RSDs) was still observed without interference from the large amount of MA. The limits of detection (LOD) of (-)-EP and (+)-pEP were approximately 2. μg/mL, therefore, their LOD as the impurity concentrations were calculated at about 0.2%. Seized MA samples were dissolved in water at a high concentration (1. mg/mL) and analyzed by this method. (-)-EP and (+)-pEP were clearly detected as impurities. Although these compounds had similar migration times and mass spectral patterns, the fine repeatability allowed easy identification of the impurities by a simple comparison of the absolute migration times of the specimens and those of authentic standards. This study is the first to report the use of a chemically modified capillary for the impurity profiling on CE/MS/MS.

Methamphetamine (MA) is the most common drug of abuse in Japan. MA is produced by chemical synthesis and final products of the drug contain small amounts of precursor chemicals, intermediates and by-products. Chirality is a principal characteristic of MA for evaluating the precursor chemicals. Adulterants such as dimethyl sulfone might be associated closely with trafficking routes of illicit drugs. Drug profiling is a scientific tool to identify the synthesis route, the sources of supply, trafficking routes and connections between seizures, which supports drug law enforcement agencies in their effort to eliminate organized drug crime. This review summarizes methods for characterization and profiling of MA hydrochloride, mainly based on our findings.

An automated full-range quantitation method for identifying d-methamphetamine (d-MA) in human hair using a microchip-based ELISA system (microELISA) in combination with a micropulverized extraction method for sample preparation has been developed. An anti-d-MA antibody and a peroxidase-linked MA were used for the competitive ELISA assay. Method validation was carried out using doped hair samples, and segmental analyses of real-case specimens were carried out by both microELISA and liquid chromatography/tandem mass spectrometry (LC-MS/MS) to verify the reliability and applicability of this new method. Sample preparation and quantitation can be accomplished easily and quickly (less than 30 min) in smallscale contamination-free environments. © 2008 CBMS.

We have developed a novel basic technology for terahertz imaging, which allows detection and identification of drugs concealed in envelopes, by introducing the component spatial pattern analysis. The spatial distributions of the targets are obtained from terahertz multispectral transilluminalion images, using absorption spectra measured with a tunable terahertz-wave source. The samples we used were methamphetamine and MDMA, two of the most widely consumed illegal drugs in Japan, and aspirin as a reference. The three kinds of samples hidden in mail envelopes were clearly distinguished. Hereby we expect this technology to contribute to the screening letters and packages. © 2003 Optical Society of America.

Classification of seized methamphetamine by impurity profiling can provide very useful information in criminal investigations of drug traffic routes, sources of supply and relationships between seizures. The aim of this study is to improve and develop an analytical method for detecting impurities such as starting materials and by-products in illegally prepared methamphetamine.HCl samples. A 50 mg sample of methamphetamine.HCl was dissolved in 1 ml of buffer solution (four parts 0.1 M phosphate buffer pH 7.0 and one part 10% Na2CO3). Impurities were extracted with 0.5 ml of ethyl acetate containing four internal standards (ISs) (n-decane, n-pentadecane, n-nonadecane and n-hexacosane) and analyzed by gas chromatography (GC) using a flame ionization detector (FID) on a DB-5 capillary column (0.32 mm i.d. x 30 m, film thickness 1.0 mum). The use of a middle-bore column offered better separation of the impurity peaks. The correction of the retention times of impurity peaks with four ISs made peak identification very accurate for subsequent data processing. Twenty-four characteristic peaks were selected for comparison and similarity and/or dissimilarity between samples, and the data were evaluated by the Euclidean distance of the relative peak areas after logarithmic transformation. The results indicate that the present method would be useful for methamphetamine impurity profiling. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved.

Benzodiazepines are one of the most widely prescribed drugs for the treatment of a wide spectrum of clinical disorders. They are used as anticonvulsants, anxiolytics, hypnotics or muscle relaxants with different duration of action. In this paper, a simple and sensitive method for the determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry (GC/MS) is described. The drugs spiked in whole blood were extracted with an Oasis(TM) HLB solid-phase extraction cartridge (Waters), which contains a copolymer designed to have a hydrophilic-lipophilic balance. GC/MS analysis was performed using a Shimadzu QP-5000 equipped with a BPX5 capillary column (15 mx0.32 mm I.D., film thickness 0.25 μm, SGE). Nineteen benzodiazepines and two thienodiazepines were well separated from each other on their SIM chromatograms and also on the TIC with the exception of oxazolam to cloxazolam separation. The blank extract from whole blood gave no peaks that interfered with all benzodiazepines and thienodiazepines on the chromatogram. The calibration curves for selected benzodiazepines with fludiazepam as an internal standard showed excellent linearity over the concentration range 5-500 ng/ml blood with a correlation coefficients of &gt 0.995. The detection limits ranged from 0.2 to 20 ng/ml blood. The method is simple and sensitive for the determination of benzodiazepines in whole blood and seems to be useful in the practice of forensic science. Copyright (C) 2000 Elsevier Science Ireland Ltd.

A column-switching liquid chromatography (LC) system combined with electrospray ionization mass spectrometry (ESI-MS) was developed for the determination of N-methyl-1(1,3-benzodioxol-5-yl)-2-butanamine (MBDB) and its metabolites in urine and serum samples. Filtered samples were injected and concentrated onto a strong cation-exchange (SCX) precolumn, which was coupled to a reversed-phase analytical column via a six-port valve. MBDB and its metabolites were separated on the analytical column using 10 mM ammonium acetate (pH 5) - acetonitrile (80:20) as a mobile phase. Analytes were eluted directly into the ESI source of ion-trap detector. Linearity was obtained over the concentration range of 5-500 ng/ml for MBDB and 20-1000 ng/ml for the metabolites. The detection limit of each compound in urine or serum samples ranged 5-20 ng/ml using scan mode. This method is rapid and sensitive for the determination of MBDB and its metabolites in biological samples.

In an attempt to provide a better understanding of the scope and limitations of animal models used in some drug development programs and to further our understanding of potential metabolic bioactivation reactions, we have undertaken studies to profile the monoamine oxidase A and B (MAO-A and -B, respectively) activities in liver and brain mitochondrial preparations obtained from a variety of species using a series of 1-methyl-4-aryl-1,2,3,6-tetrahydropyridinyl substrates. Mitochondrial preparations were incubated with substrates at 37 degrees C in the presence or absence of clorgyline, (R)-deprenyl, or a mixture of these two propargylamines to inhibit MAO-A, MAO-B, or both enzymes. The rates of formation of the corresponding dihydropyridinium metabolites were estimated spectrophotometrically. MAO-B was found to be the principal enzyme present in all tissues. Human liver displayed more MAO-A activity than the liver of any other species studied; subhuman primates displayed little or no detectable MAO-A activity. The properties of the preparations from rat liver were most similar to those from human liver with respect to the MAO-A/MAO-B ratios and the kinetic parameters of the four substrates used to profile enzymatic activity. The kinetic properties of mitochondrial preparations from bovine liver, a commonly used source of purified MAO-B preparations, were consistently different from all of the other species studied. The mitochondrial preparations from rabbit brain and liver also were unusual in that they displayed relatively low MAO activities. Additionally, these enzyme activities were considerably less susceptible to inhibition by clorgyline and (R)-deprenyl. Finally, an exceptionally low MAO-B liver/brain V-max/K-m ratio was observed with the mitochondria obtained from the C57BL/6 mouse, an effect that may contribute to the susceptibility of this strain to the toxic effects of the parkinsonian-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.

The metabolism of N-Methyl-1-(1,3-Benzodioxol-5-yl)-2-Butanamine (MBDB) in rat was investigated. MBDB hydrochloride was administrated orally to male Wistar rats at a dose of 20mg/kg. After enzymatic hydrolysis, the urine were extracted with chloroform-isopropanol (3:1, v/v) under alkaline (pH9) condition. The extracts were analyzed by gas chromatography-mass spectrometry (GC-MS) after trifluoroacetyl (TFA) -derivatization using MBTFA. MBDB, 1-(1,3-benzodioxol-5-yl)-2-butanamine (BDB), N-methyl-1-(4-hydroxy-3-methoxyphenyl)-2-butanamine and 1-(4-hydroxy-3-methoxyphenyl)-2-butanamine were identified as urinary metabolites.

A reliable and sensitive method for the determination of triazolam in human muscle using gas chromatography-mass spectrometry (GC-MS) is described. The drug was extracted from decomposed human muscle using three-step liquid-liquid extraction and HPLC which was performed isocratically on a conventional ODS column with a mobile phase of 0.01 M phosphate buffer (pH 6.5)-acetonitrile (7:3). Estazolam was used as an internal standard. GC-MS analysis was performed on a DB-5 capillary column. Excellent linearity was obtained over the concentration range 1-200 ng/g. The lower limit of detection was approximately 0.5 ng/g. Forensic application of the present method was also described. (C) 1997 Elsevier Science B.V.

This paper describes a simple and sensitive high-performance liquid chromatographic (HPLC) method for the detection of human globin chains in blood and bloodstains. The method involves direct injection of the filtered samples of dilute hemolysates or bloodstain extracts onto a microbore C-4 reversed-phase column (2.1 mm I.D.) with UV detection at 220 nm. Microbore HPLC offers a significant improvement in sensitivity with little loss of the resolution of globin chains and only small variations in the determination of gamma chain composition. The detection limit of hemoglobin (Hb) was 0.1 mu g, which is equivalent to about 1 nl of fresh whole blood. Umbilical cord blood could be differentiated from adult blood in stains that were up to twenty weeks old, by the presence of gamma globin chains. The present method will be useful for detection of abnormal Hbs and for the determination of gamma chain composition in clinical laboratories, as well as in the practice of forensic science for the analysis of minute amounts of blood and bloodstains.

A 24-year-old male was found dead in a car left in a river for about 3 months. The cadaver was almost adipoceratous and autopsy findings revealed that there were neither remarkable injuries nor lethal diseases. Toluene, ethano, 1-propanol, 2-propanol, 1-butanol, dimethyl sulfide, dimethyl disulfide, isovaleraldehyde and n-butyl n-butyrate were detected in the specimens collected at the autopsy by head space gas chromatography/mass spectrometry (GC/MS). The toluene concentrations (mu g/g) were 31.0 in brain, 10.6 in liver, 5.4 in kidney, 15.0 in skeletal muscle and 187.1 in adipose tissue. The presence of diatom in lung, liver and kidney suggested that death was caused by drowning. So far as we know, this is the first report of detection of toluene in an adipoceratous body.