Determination of bromazepam, clonazepam and metabolites after a single intake in urine and hair by LC-MS/MS. Application to forensic cases of drug facilitated crimes

The number of reports on drug facilitated crimes is increasing these last years. Apart from ethanol and cannabis, benzodiazepines (BZD) and analogs are the most common drugs reported to be used probably due to their amnesic and sedative properties. We have developed a rapid and sensitive method using LC-MS/MS triple stage quadrupole (TSQ) for the determination of single exposure to bromazepam (Lexomil, 6 mg) and clonazepam (Rivotril, 2 mg) in urine and hair of healthy volunteers. Chromatography was carried out on a Uptisphere ODB 5 microm, 2.1 mm x 150 mm column (Interchim) with a gradient of acetonitrile and formate 2 mM buffer, pH 3. Urine was extracted with Toxitube A (Varian) and allowed the detection of bromazepam, 3-hydroxy-bromazepam, clonazepam and 7-Aminoclonazepam for more than 6 days. Head hair, collected 1 month after the exposure, was treated by incubation with Soerensen buffer pH 7.6, followed by liquid-liquid extraction with dichloromethane for common BZD. A specific pre-treatment for amino-BZD, with an incubation of 15 min at 95 degrees C in 0.1 N NaOH before liquid-liquid extraction with dichloromethane, gave better recoveries and repeatability. After single exposure, bromazepam was present in powdered hair at 28 pg/mg and 7-Aminoclonazepam at 22 pg/mg in the first 1-cm segment, while no clonazepam was detectable. This method was applied in two forensic cases. It allowed us to determine bromazepam in urine 3 days after the alleged offense and in cut head hair at a concentration of 6.7 pg/mg only in the 2-cm proximal segment. The other case showed the presence of clonazepam and 7-Aminoclonazepam in urine a few hours after the offense and the presence of 7-Aminoclonazepam at about 3.2 pg/mg in axillary hair 4 months later.     

Preparation of degraded human DNA under controlled conditions

DNA typing through analysis of short tandem repeats (STRs) and mitochondrial DNA (mtDNA) by means of the polymerase chain reaction (PCR) and sequencing are the common methods for the forensic identification of persons and reconstruction of kinship, especially when skeletal human remains have to be analyzed. Furthermore, samples typically found at crime scenes may be both quantitatively and qualitatively inadequate since they may contain very scarce and often degraded DNA due to exposure to heat, light, humidity, and microorganisms. In order to improve the performance of STR typing technology in those cases where DNA availability is limited, it would be desirable to have a source of degraded DNA with known properties. For this purpose, we have developed a method to prepare artificially degraded DNA under controlled conditions. By treatment of genomic DNA with sonication and DNAse I we have produced DNA fragments within a defined range of lengths. STR typing of this degraded DNA with a commercially available multiplex kit could only produce partial profiles as indicated by the absence of STR alleles with sizes >200 bp. This artificially degraded DNA can be used for the improvement and standardization of STR typing protocols when only highly degraded DNA is available for analysis.     

Particle size analysis of soils under simulated scene of crime conditions: the interest of multivariate analyses

Two simulated scenes of crime have been studied. Soil traces adhering to boots, sport shoes and tissues have been compared with control samples using particle size analysis. Comparisons of percentage of particles per class interval and multivariate analyses were used to determine how the size distribution of each suspect sample varied compared to the original distribution. A loss of coarse fractions characterized most of the suspect samples even though they were derived from soils having different distributional patterns. However, the differences between control and suspect samples were found to be more important for soils dominated by extreme size classes than for soils rich in medium size particles. The differences among soil samples from the same source were found to be restricted to the extreme size classes. The finding of significant differences within size classes ranging from 1 to 0.063 mm should be then considered as a dissimilarity sign between trace sample and hypothetic control soil.     

The reactivity of gamma-hydroxybutyric acid (GHB) and gamma-butyrolactone (GBL) in alcoholic solutions

In this work the stability of GBL (gamma-butyrolactone) and GHB (gamma-hydroxybutyric acid) in alcoholic media was studied. Under acidic conditions the GBL will react with ethanol or methanol to give the corresponding ethyl and methyl esters of GHB. It can be seen that ester formation is dependent on the type of alcohol, the alcohol content of the solution, and the pH of the solution. Under the same conditions it was shown that GHB does not give rise directly to the corresponding ester when merely in the presence of an alcohol; however the ester will be formed if the conditions are present for conversion of GHB to GBL followed by subsequent reaction with alcohol. In alcoholic beverage samples spiked with GBL the expected conversion to GHB occurred, and the formation of the ethyl ester of GHB was also seen in some samples. Wine samples were analyzed for the presence of the ethyl ester of GHB, and the effect of adding GHB/GBL to hot beverages was studied.     

Domestic violence in Puerto Rican gay male couples: perceived prevalence, intergenerational violence, addictive behaviors, and conflict resolution skills

Domestic violence (DV) is a pattern of behaviors in the context of an intimate relationship, which can be manifested in emotional, physical, or sexual abuse. DV currently represents a social and a public health issue. This study is an effort to foster a better understanding of DV among same-sex couples. In it, the authors included the participation of 199 Puerto Rican gay males to identify prevalence of DV, violence in their family of origin, participants' addictive behaviors and exposure to violence at childhood, and their conflict resolution skills. Participants were relatively young, highly educated Puerto Rican gay men who reported a high level of domestic violence in their relationships. This violence was identified as emotional violence by 48% of the participants. This sample reported high levels of violence among their families of origin and low conflict resolution skills.     

AIDS in the workplace: a program that's still relevant

A new AIDS-in-the-Workplace program being implemented in Québec emphasizes the rights of workers living with HIV/AIDS. The program tackles issues such as confidentiality of HIV status, access to insurance benefits, discrimination, and the need for employers to take reasonable measures to accommodate the disabilities of workers.     

Prevalence and Processing of Child Sexual Abuse: A Multi-Data-Set Analysis of Urban and Rural Counties

This study investigated how contextual factors affect the processing of child sexual abuse cases, from reporting to sentencing. We analyzed three types of data: (a) data compiled by the Pennsylvania Coalition Against Rape from monthly reports by all rape crisis centers in the state; (b) data from the Pennsylvania Office of Children, Youth, and Families; and (c) sentencing data from the Pennsylvania Commission on Sentencing. Results indicated that aggregate rates of reporting, substantiation, and sentencing were affected by county levels of expenditure. In addition, reporting and substantiation were affected by county-level factors, such that rural counties, counties with a higher percentage of individuals living below the poverty level, counties with higher expenditures, and counties with a higher percentage of stranger assaults had higher rates of child sexual abuse reporting.     

Bone as a biomarker of acute fluoride toxicity

The use of bone as a biomarker for chronic and acute exposure to fluoride salts has been suggested, but there are no data published about its use to evaluate lethal intoxication. One hundred and sixty rats were divided into eight groups that received a single oral intubation dose from 0 (control) to 90 mg F/kg as NaF. The animals' time of death was recorded and their femurs were removed for fluoride analysis. Acid-soluble fluoride was determined in the whole bone and on the surface (periosteal), using an ion specific electrode. The data showed a statistically significant relationship between fluoride dose and the number of deaths (P<0.0001). A statistically significant relationship was also found between fluoride dose and fluoride concentration ([F]) in either the whole femur (P<0.0017), on the surface (P<0.0001) or for the ratio periosteal [F]/whole [F] (P<0.0001). However, the [F] on the femur surface was more closely correlated with mortality than that in the whole bone, showing statistically significant differences among the lethal doses and control (P<0.05). The data suggest that the ratio [F] periosteal bone/[F] whole bone, is a biomarker for acute fluoride toxicity.