Validation of an LC–MS Method for the Detection and Quantification of BZP and TFMPP and their Hydroxylated Metabolites in Human Plasma and its Application to the Pharmacokinetic Study of TFMPP in Humans*

Abstract: An LC–MS method was developed for benzylpiperazine (BZP) and trifluoromethylphenylpiperazine (TFMPP), constituents of “party pills” or “legal herbal highs,” and their metabolites in human blood plasma. Compounds were resolved using a mixture of ammonium formate (pH 4.5, 0.01 M) and acetonitrile (flow rate of 1.0 mL/min) with a C18 column. Calibration curves were linear from 1 to 50 ng/mL (R² > 0.99); the lower limit of quantification (LLOQ) was 5 ng/mL; the accuracy was >90%; the intra‐ and interday relative standard deviations (R.S.D) were <5% and <10%, respectively. Human plasma concentrations of TFMPP were measured in blood samples taken from healthy adults (n = 6) over 24 h following a 60‐mg oral dose of TFMPP: these peaked at 24.10 ng/mL (±1.8 ng/mL) (C_max) after 90 min (T_max). Plasma concentrations of 1‐(3‐trifluoromethyl‐4‐hydroxyphenyl) piperazine peaked at 20.2 ng/mL (±4.6 ng/mL) after 90 min. TFMPP had two disposition phases (t_½ = 2.04 h (±0.19 h) and 5.95 h (±1.63 h). Apparent clearance (Cl/F) was 384 L/h (±45 L/h).     

Evaluation of some oxygen, sulfur, and selenium substituted ninhydrin analogues, nitrophenylninhydrin and benzo[f]furoninhydrin

Six ninhydrin analogues containing oxygen, sulfur, and selenium substituents at the C-5 position, 5-(4-nitrophenyl)ninhydrin, and benzo[f]furoninhydrin were evaluated as fingerprint development reagents. The analogues all showed good fingerprint color development but were not superior to ninhydrin in this respect. The benzo[f]furoninhydrin complex was strongly luminescent at room temperature following zinc complexation, while the remaining analogues required cooling to -196 degrees C to produce optimum luminescence. The benzo[f]furo, nitrophenyl, and methyl selenide analogues showed the best potential as fingerprint reagents with the benzo[f]furo analogue comparing favorably with DFO.     

An improved method for post-PCR purification for mtDNA sequence analysis

Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.     

Exploring gender differences in dating violence/harassment prevention programming in middle schools: results from a randomized experiment

In this study, we randomly assigned 123 sixth and seventh grade classrooms from seven middle schools in the greater Cleveland area to one of two five-session curricula addressing gender violence/sexual harassment (GV/SH) or to a no-treatment control group. A baseline survey and two follow-up surveys were administered immediately after the treatment (Wave 2) and about six months post-treatment (Wave 3). In an earlier paper, we demonstrated the effectiveness of two approaches to youth GV/SH prevention programming (a fact-based, law and justice curriculum and an interaction-based curriculum). In this paper, we explored whether these largely positive findings remain for both girls and boys, including whether girls experience higher levels of GV/SH than boys. Most of our statistical models proved to be non-statistically significant. However, in 2 of our 48 victimization/perpetration (any violence, sexual violence and non-sexual violence) models (across two post-intervention follow-up points), we observed that the interventions reduced peer (male or female, non-dating partner) sexual violence victimization and reduced peer perpetration, but another outcome model indicated that the interventions increased dating perpetration. These mixed findings will need to be explored further in future research. Regarding our primary research question, we observed no statistically significant differences for the treatment multiplied by gender interaction terms for any of the perpetration or victimization outcome models, suggesting that the treatment had similar effects on girls and boys. However, we did observe that boys are more involved in violence than girls: both as victims and perpetrators. Boys experienced significantly more of three types of victimization from peers and dating partners compared to what girls experienced at the hands of their peers and dating partners. As perpetrators, boys committed more sexual victimization against peers (immediately post-intervention only) and more sexual victimization against dating partners than girls. The implications of these results are discussed.     

Object-oriented Bayesian networks for paternity cases with allelic dependencies

This study extends the current use of Bayesian networks by incorporating the effects of allelic dependencies in paternity calculations. The use of object-oriented networks greatly simplify the process of building and interpreting forensic identification models, allowing researchers to solve new, more complex problems. We explore two paternity examples: the most common scenario where DNA evidence is available from the alleged father, the mother and the child; a more complex case where DNA is not available from the alleged father, but is available from the alleged father’s brother. Object-oriented networks are built, using HUGIN, for each example which incorporate the effects of allelic dependence caused by evolutionary relatedness.     

The psychological effects of solitary confinement on prisoners in supermax units: reviewing what we know and recommending what should change

This article examines the psychological consequences of short- and long-term solitary confinement for prisoners in the United States subjected to administrative or disciplinary segregation. Particular attention is paid to the use of secure housing units, alternatively known as control units or supermax units. These correctional entities allow for the isolation of convicts under conditions that offer little sensory stimulation and minimal opportunities for interaction with other people. The circumstances typically found in these units and the heightened potential for the abuse of prisoners are described. The connections between internment and mental illness-as well as isolation and race, gender, and class-are explored. A set of recommendations for the reform of secure housing is presented.     

Joint match probabilities for Y chromosomal and autosomal markers

Empirical tests of association between Y chromosome and autosomal markers are presented and a theoretical framework for determining a joint match probability is recommended. Statistical analyses of association were performed in 16 US populations between the autosomal genotypes from loci CSF1PO, FGA, THO1, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S512, D21S11 and Y chromosome haplotypes from loci DYS19, DYS385ab, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439. The sample populations include individuals of European-, African-, Hispanic-, Native-, and Asian-American ancestry. The results are consistent with independence of Y and autosomal markers, although small amounts of dependence would likely have escaped our tests. Given the data in hand, we suggest it is appropriate to compute joint match probabilities by multiplying the Y haplotype frequency with the appropriately corrected autosomal frequency. In addition to correcting for autosomal frequency differences between groups, a further correction may be required. Since two individuals sharing the same Y haplotype are likely to be more recently related than two randomly chosen individuals, the autosomal frequencies have to be adjusted to account for this, akin to the theta correction used to account for population substructure. The structure imposed on the autosomal frequencies conditioned in a Y match is a function of the number of markers scored and their mutation rate. However, in most settings theta<0.01. When population structure is already present in the autosomes, the additional effect due to conditioning on the Y is small. For example, if the amount of structure in the population is theta=0.01 or 0.03 (the NRCII range), then the effect of conditioning on the Y results in only a trivial increase in theta to 0.02-0.04, respectively.     

Y-chromosome STR system, Y-PLEX 12, for forensic casework: development and validation

The Y-PLEX 12 system, developed for use in human identification, enables simultaneous amplification of eleven polymorphic short tandem repeat (STR) loci, namely DYS392, DYS390, DYS385 a/b, DYS393, DYS389I, DYS391, DYS389II, DYS 19, DYS439 and DYS438, residing on the Y chromosome and Amelogenin. Amelogenin provides results for gender identification and serves as internal control for PCR. The validation studies were performed according to the DNA Advisory Board's (DAB) Quality Assurance Standards. The minimal sensitivity of the Y-PLEX 12 system was 0.1 ng of male DNA. The mean stutter values ranged between 3.76-15.72%. A full male profile was observed in mixture samples containing 0.5 ng of male DNA and up to 400 ng of female DNA. Amelogenin did not adversely affect the amplification of Y-STRs in mixture samples containing male and female DNA. The primers for the Y-STR loci present in Y-PLEX 12 are specific for human DNA and some higher primates. None of the primate samples tested provided a complete profile at all 11 Y-STR loci amplified with the Y-PLEX 12 system. Y-PLEX 12 is a sensitive, valid, reliable, and robust multiplex system for forensic analysis, and it can be used in human forensic and male lineage identification cases.     

The evaluation of the extent of transporting or "tracking" an identifiable ignitable liquid (gasoline) throughout fire scenes during the investigative process

Tests have determined that boots or shoes of individuals at a fire scene do not transport sufficient contaminants ("tracking") through the fire scene to produce a positive laboratory result for the presence of gasoline in a fire scene that was not present at the time of the fire. Questions about the validity of forensic laboratory results have been raised on the basis that low-level gasoline residues detected in the laboratory samples could have been the result of transporting the residue by footwear contaminated from the fire scene ("tracking"). The data collected in this study establish that "tracking" does not lead to false-positive laboratory results. Canines trained and experienced in the detection of trace ignitable liquid residues were also utilized in this study. The canine results confirmed that properly trained canines show a higher sensitivity than do standard ASTM laboratory techniques for fire debris analysis. In a few cases, canines responded to contamination, but laboratory testing (which is the definitive indicator) did not produce positive results.     

Suspect/Foil Identification in Actual Crimes and in the Laboratory: A Reality Monitoring Analysis

Four reality monitoring variables were used to discriminate suspect from foil identifications in 183 actual criminal cases. Four hundred sixty-one identification attempts based on five and six-person lineups were analyzed. These identification attempts resulted in 238 suspect identifications and 68 foil identifications. Confidence, automatic processing, eliminative processing and feature use comprised the set of reality monitoring variables. Thirty-five verbal confidence phrases taken from police reports were assigned numerical values on a 10-point confidence scale. Automatic processing identifications were those that occurred “immediately” or “without hesitation.” Eliminative processing identifications occurred when witnesses compared or eliminated persons in the lineups. Confidence, automatic processing and eliminative processing were significant predictors, but feature use was not. Confidence was the most effective discriminator. In cases that involved substantial evidence extrinsic to the identification 43% of the suspect identifications were made with high confidence, whereas only 10% of the foil identifications were made with high confidence. The results of a laboratory study using the same predictors generally paralleled the archival results. Forensic implications are discussed.