Improved gas chromatography-negative ion chemical ionization tandem mass spectrometric method for determination of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid in hair using mechanical pulverization and bead-assisted liquid-liquid extraction

A gas chromatography-negative ion chemical ionization tandem mass spectrometric (GC-NCI-MS/MS) method was developed and validated for the determination of 11-nor-Δ(9)-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in human hair. After decontamination, hair samples were weighed (25mg), mechanically pulverized with a bead mill, and incubated in 0.7 mL of 1.0M sodium hydroxide at 95 °C for 30 min. Bead-assisted liquid-liquid extraction was performed with n-hexane:ethyl acetate (9:1, v/v), a method developed in our laboratory. The extract was evaporated to dryness, derivatized with pentafluoropropanol and pentafluoropropionic anhydride, and analyzed by GC-MS/MS in the negative ion chemical ionization mode using methane as the reagent gas. The linear ranges were 0.05-10.0 pg/mg for THC-COOH with the coefficient of determination (r(2) = 0.9976). The intra-day and inter-day precisions were within 1.7 and 13.8%, respectively. The intra-day and inter-day accuracies were -4.8 to 10.0% and -3.9 to 3.8%, respectively. The limit of detection and quantification were 0.015 and 0.05 pg/mg, respectively. The recoveries were in the range of 79.4-87.1%. The results indicate that the proposed method is simple, rapid, accurate, and precise for determination of THC-COOH in hair. The method identified THC-COOH in hair specimens from suspected marijuana abusers.     

Establishment of the measurement uncertainty of 11-nor-D9-tetrahydrocannabinol-9-carboxylic acid in hair

The quantitative analysis of 11-nor-D(9)-tetrahydrocannabinol-9-carboxylic acid (THCCOOH) in hair requires a sensitive method to detect a low-pg level. Before applying the method to real hair samples, the method was validated; in this study, we examined the uncertainty obtained from around the cut-off level of THCCOOH in hair. We calculated the measurement uncertainty (MU) of THCCOOH in hair as follows: specification of the measurand, identification of parameters using "cause and effect" diagrams, quantification of the uncertainty contributions using three factors, the uncertainty of weighing the hair sample, the uncertainty from calibrators and the calibration curve, and the uncertainty of the method precision. Finally, we calculated the degrees of freedom and the expanded uncertainty (EU). The concentration of THCCOOH in the hair sample with its EU was (0.60 ± 0.1) × 10(-4)ng/mg. The relative uncertainty percent for the measurand 0.60 × 10(-4)ng was 9.13%. In this study, we also selected different concentrations of THCCOOH in real hair samples and then calculated the EU, the relative standard uncertainty (RSU) of the concentration of THCCOOH in the test sample [u(r)(c0)], the relative uncertainty percent, and the effective degree of freedom (v(eff)). When the concentrations of THCCOOH approached the cut-off level, u(r)(c0) and the relative uncertainty percent increased but absolute EU and v(eff) decreased.     

A study on the concentrations of 11-nor-Δ(9)-tetrahydrocannabinol-9-carboxylic acid (THCCOOH) in hair root and whole hair

In this study, we investigated the patterns of cannabis users (n=412) according to their sex, age, and the results of urinalysis and hair analysis, and classified the concentrations of THCCOOH in hair into three categories to examine the levels of cannabis use. We also compared the concentrations of THCCOOH in hair root, hair without the hair root and whole hair and examined the relationship among them according to the results of urinalysis. The hair samples were washed, digested with 1ml of 1M NaOH at 85°C for 30min and extracted with 2ml of n-hexane:ethyl acetate (9:1) two times after adding 1ml of 0.1N sodium acetate buffer (pH 4.5) and 200μl of acetic acid. The final mixture was derivatized with 50μl of PFPA and 25μl of PFPOH for 30min at 70°C. The solution was evaporated, and the residue was reconstituted in 40μl of ethyl acetate and transferred to an autosampler vial. One microlitre was injected into the GC/MS/MS-NCI system. The concentrations of THCCOOH ranged from 0.06 to 33.44pg/mg (mean 2.96; median 1.32) in hair from cannabis users who had positive urine results and ranged from 0.05 to 7.24pg/mg (mean 1.35; median 0.37) in hair from cannabis users who had negative urine results. The average concentration of THCCOOH in hair from cannabis users who had positive urine results was higher than that from cannabis users who had negative urine results. Male cannabis users in their forties were predominant. We classified the concentrations of THCCOOH in hair into three groups (low, medium and high), and could use the grouping of THCCOOH in hair as a guide for determining the level of use. The low, medium and high concentration ranges for THCCOOH in hair were 0.05-0.24, 0.25-2.60 and 2.63-33.44pg/mg, respectively. We also investigated 28 hair samples with the root. The highest concentrations of THCCOOH were seen in the hair root from 18 out of the 28 hair samples. The average concentrations of THCCOOH in hair root, hair without hair root and whole hair from cannabis users who had positive urine results were higher than those who had negative urine results.     

How Concessional is Aid Lending?

The method used by Development Assistance Committee countries for measuring the concessionality of aid loans has remained unchanged for nearly 20 years. It was designed to measure the net cost of aid to donors not the net benefit to recipients. The discount rate used takes no account of changes in the value of the currency of the loan or of changes in prices for goods traded by recipient countries. Furthermore, it does not consider the implications of tying of aid or of policy conditionality. This paper suggests an alternative measure that shows the real net benefit of aid finance to recipients. It argues that the discount rate used by the Development Assistance Committee is too high and that changes in the value of the currency in which a loan is taken out can be important. Nevertheless, real rates of interest for developing countries remain surprisingly high despite low nominal rates due to falling prices of traded goods. This finding has implications for the future real cost of debt service to recipients.     

The Impact of Radical Right-Wing Parties on the Positions of Mainstream Parties Regarding Multiculturalism

What are the political effects of rising radical right-wing parties (RRPs) in Western Europe? Does the rise of the parties drive mainstream parties (MPs) to become more restrictive on issues mobilised by RRPs, such as multiculturalism? Analysing manifesto data from 1981 to 2008, it is found that the rise of RRPs makes right-wing MPs adopt more restrictive positions regarding multiculturalism. However, left-wing MPs do so only when the opinion of party supporters on foreigners becomes more negative or when the parties lost more votes in the previous election than their opponent right-wing MPs did. The result implies that niche parties with extremist positions can benefit from their own electoral success by dragging MPs toward their own positions. However, the impact of rising niche parties on MPs should be understood against a broader background of party competition, and the impact can be dissimilar between MPs with different ideological commitment and strategic opportunities.     

Developmental patterns of sex differences in delinquency among African American adolescents: A test of the sex-invariance hypothesis

This paper addresses a developmental issue concerning longitudinal patterns of sex differences in delinquency. Hirschi and Gottfredson argue that the age-delinquency relation is invariant across sex and that sex differences in delinquency are invariant over time as well. A combination of these two propositions generates a hypothesis, called here the sex-invariance hypothesis, that sex differences in delinquency are invariant over developmental stages of adolescents. To test the sex-invariance hypothesis, nine waves of panel data collected from a representative urban sample of African American adolescents are analyzed. The overall findings show that sex differences in delinquency tend to vary as the subjects grow older, rather than remain constant as the invariance thesis posits. Specifically, sex differences in delinquency peak at the age of 15 and thereafter declines with age. We also find that parental supervision significantly explains sex differences in delinquency for younger adolescence, but not for older adolescence.     

The application of miniplex primer sets in the analysis of degraded DNA from human skeletal remains

A new set of multiplexed PCR primers has been applied to the analysis of human skeletal remains to determine their efficacy in analyzing degraded DNA. These primer sets, known as Miniplexes, produce shorter amplicons (50-280 base pairs (bp)) than standard short tandem repeat (STR) kits, but still utilize the 13 CODIS STR loci, providing results that are searchable on national DNA databases. In this study, a set of 31 different human remains were exposed to a variety of environmental conditions, extracted, and amplified with commercial and Miniplex DNA typing kits. The amplification efficiency of the Miniplex sets was then compared with the Promega PowerPlex 16 system. Sixty-four percent of the samples generated full profiles when amplified with the Miniplexes, while only 16% of the samples generated full profiles with the Powerplex 16 kit. Complete profiles were obtained for 11 of the 12 Miniplex loci with amplicon sizes less than 200 bp. These data suggest smaller PCR amplicons may provide a useful alternative to mitochondrial DNA for anthropological and forensic analysis of degraded DNA from human skeletal remains.     

Use of hand-held laser scanning in the assessment of craniometry

The intra- and inter-examiner reliability was evaluated for hand-held 3D laser scanning technology when it was combined with localization of landmarks for craniometry. The data from the laser surface scanning were compared with those of conventional direct measuring. Using thirty unidentified skulls requested for individual identification, measurements were taken of the line distances from lambda to 26 landmarks, and also for seven breadth parameters. For the laser surface scanning, two examiners performed replicate measurements with an interval of 1 week. In the conventional direct measuring, the first examiner took replicate measurements with a 1-week interval. To assess intra- and inter-examiner reliabilities, the intraclass correlation coefficient was used. Analysis of variance with repeated measures for each parameter was performed to compare the conventional method with the 3D scanning method. Both the 3D scanning and conventional methods showed excellent intra-examiner reliabilities, and the 3D laser scanning method also showed excellent inter-examiner reliability. A statistical difference between the two examiners was found only in nasal breadth in the 3D laser scanning method. There was no significant difference between the two measuring methods, though the 3D laser scanning method tended to give a slightly lower reading. Collectively, the 3D laser scanning method with point localization is a useful method with excellent reliability, and it can replace the conventional direct measuring method in craniometry.     

Standardization of method for the analysis of dextromethorphan in urine

Dextromethorphan (DMP), an antitussive, is one of the most popular drugs among the younger generation in Korea. It usually is taken for its hallucinogenic properties and overdoses have been responsible for the fatalities that have been reported frequently. To control the abuse of DMP, the authorities restricted its use through classifying it as a controlled drug on October 2003. The purpose of this study is to provide a standard method for the analysis of DMP and its main metabolite, dextrorphan (DTP) in biological specimens. At first we established a standard operating procedure (SOP) for DMP/DTP in urine, and a method validation was performed. We also quantified DMP from 16 drug abuser's urine samples all of which were positive in the screening test for DMP. For the detection of DMP/DTP, urine samples were adjusted with 6N NaOH (pH 11) and extracted with ethylacetate. Thin layer chromatography was used as the screening test, and the final identification for DMP/DTP was used by GC/MS. The ions (m/z 271 for DMP, m/z 257 for DTP and m/z 86 for lidocaine as internal standard) were extracted from the full scan mass spectrum and were used for quantification. The selectivity, linearity of calibration, accuracy, within- and between day precision, limit of detection and quantification, recovery and stability were examined as parts of the method validation. Extracted calibration curves were linear from 100 to 2000 ng/mL for DMP and DTP with correlation coefficients better than 0.999. Limit detection was 50 ng/mL for DMP and DTP. Within-run precision (%CV) for DMP and DTP at three different concentrations (100, 500 and 1000 ng/mL) was 6.10-18.85%, and between-run precision was 1.70-7.86% for DMP and DTP. Absolute recovery for DMP and DTP was 57-74%, and relative recovery (extraction efficiency) was 80-89%. For 16 drug abuser's urine samples, the concentrations of DMP and DTP were 0.16-52.63 and 0.41-23.75 microg/mL, respectively. Method validation is an important requirement in the practice of chemical analysis, and it will be particularly useful in verifying the reliability of analytical results in the field of forensic science.