Development of analytical method for illegal substances in sweat and comparison of the effectiveness of sweat collection materials.

PURPOSE: Sweat analysis can yield valuable information in forensic investigations, diagnosis and treatment. The purpose of this study was to develop a validated gas chromatography-mass spectrometric method for the detection of illegal substances in sweat after optimizing the method by chemometric approach. This study also investigated the effectiveness of alternative sweat-collecting materials. METHODS: Plackett-Burman screening design was employed to determine the effect of seven process factors on this new method. Then, central composite design (CCD) was used to optimize the method. The method was validated according to the international guidelines. The effectiveness of alternative sweat-collecting materials (cosmetic pads and swabs) was compared with a commercially available collecting device (DrugWipe5A). RESULTS: Sample pH, ultrasonic bath time, and liquid-liquid extraction (LLE) shaking time were determined as the most significantly effective three parameters with the Plackett- Burman screening design. The validation procedure was successfully performed after optimizing this method. The comparison study demonstrated that cosmetic pads, swabs, and DrugWipe5A can be used interchangeably. CONCLUSIONS: Our results suggested that the statistical optimum strategy was an effective tool for the optimization of process parameters. Combined with the sensitivity and selectivity of our method, the analysis of sweat collection materials proved to be a useful tool for physicians and health care professionals.

Toxicological evaluation of alcohol and substance abuse in children and adolescents.

Unhealthy behaviors such as use of alcohol and drug usually begin during adolescence. Izmir is on transit route for illicit substance due to geographical situation. Children and adolescents are the most important threatened group in terms of alcohol and substance abuse. In this study, it was aimed to investigate alcohol and substance use profile of children and adolescents in Izmir/Turkey with the toxicological analysis results obtained from Addiction Toxicology Laboratory.Urine and blood samples of 4524 cases at and under the age of 18 years coming from various departments to the laboratory in 2015-2016 were analyzed by enzymatic immunoassay. Information and analysis results of the cases were obtained by retrospective analysis of the hospital system.83,3% of the cases were male and the mean age was 16,69 ± 1,63. Alcohol and/or substance use was determined in 13,2% of the cases. Among the cases with positive results of analysis, cannabis (33%) was mostly detected and was followed by amphetamine type stimulants (ATS, 15%), polysubstance use (15%) and alcohol (13%). While cannabis, polysubstance use and ATS were the most common in male, ethyl alcohol, ATS and benzodiazepine were mostly detected in female. There was a significant increase in the substance use rate in 2016 compared to the previous year.A substance use profile was obtained through drug testing in adolescents who are in the risk group for substance use. In this context, our data will be indicative for the development of new and more effective preventive strategies targeting children and adolescents.

The impact of creatinine reference value: Normalization of urinary drug concentrations.

Many illicit drug users attempt to manipulate urine drug testing; dilution is one of the methods. In screening tests, false-negative results below the cut-off values can create positive results after creatinine normalization. This study aimed to evaluate the impact of a creatinine reference value on the normalization of the drug concentration in diluted urine. The study focused on 25 630 cases and the following information: gender, age, urine collection time, drug screening test results, creatinine concentration (CR), and confirmation analysis result. Mean CR value was 143.71 ± 83.68 mg/dl. There was a significant difference between CR and gender (p = 0.03). The mean CR for women was lower than that for men. The correlation between age and CR was not significant (r = -0.08, p = 0.00). However, after grouping the sample into age groups of 10 years, there was a significant difference between age groups and mean CR (p = 0.00). The mean CR was significantly lower in the 0-9 year age group (n = 34) than in the 20-29 year age group (n = 10 943). According to the urine specimen collection time, CR levels during the early hours of the day (06:00-06:59) were lower than those during the remaining hours (p = 0.00). The highest converted drug-negative to drug-positive results were obtained from 153.23 mg/dl CR reference value. Reference CR values were evaluated according to gender, age, and urine collection time. Different rates of positive results were obtained for each reference value. There is no published local creatinine value for spot urine samples in many countries, including Turkey. This will be useful to develop appropriate normalization models when reporting drug test results.

Urine drug-testing tampering approaches: Turkish probationers.

The growing numbers of individual and social problems associated with drug abuse necessitate new approaches in drug-testing systems. Equally, drug abusers may attempt to invalidate drug testing using different methods such as adulteration, dilution and substitution. This study aims to investigate tampering methods commonly used by Turkish substance-using probationers and evaluate their effects on toxicological drug-testing results. Initially, probationer urinary screening test results and laboratory substitution documents were evaluated to investigate the dilution and substitution attempt. Additionally, an experimental study was carried out by using readily available household products (bleach, vinegar, drain opener, eye drops) for adulteration. The effect of these agents was investigated for 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH), amphetamine and 3,4-methylenedioxymethamphetamine (MDMA). It was determined that probationers preferred unbranded products (syringes, nylon bottles, etc.) for urine substitution. To detect dilution, screening test results were evaluated along with creatinine values. The variability of mean creatinine values can change the rate of the before-negative and after-positive ratio. For adulteration method, the high amounts of bleach provided false-negative results for THC-COOH and amphetamine, but spiking in any concentration of bleach affected MDMA results, causing a slight increase. Vinegar did not affect the THC-COOH and amphetamine results. However, false-negative results were observed for MDMA, with high amounts of vinegar-spiked urine samples. Drain opener was added in large quantities, and false-negative results were observed for all analytes. Visine eye drops did not have any effect on THC-COOH or amphetamine, but a high quantity of eye drops had a slight decreasing effect for MDMA.

New Psychoactive Substance 5-MeO-MiPT In vivo Acute Toxicity and Hystotoxicological Study.

BACKGROUND: The hallucinogenic tryptamine analog 5-methoxy-N-methyl-N-isopropyltryptamine (5-MeO-MiPT) causes social problems worldwide. There are several studies on the metabolism; however, not more studies were found in the literature on acute toxicity. AIMS: To report the acute toxicity of 5-MeO-MiPT in mice, followed by quantitative toxicological analysis of blood and organs, hystotoxicological and immunohistochemical analysis of tissues and cells. STUDY DESIGN: Animal experiment Methods: In vivo experiments were performed using CD1 adult female mice (n=26). Animals were caged in 4 groups randomly. First group was a control (n=3). Second group was vehicle control (n=3) and injected 150 µL of blank solution (50% dimethyl sulfoxide in saline /0.9% of NaCl). While for acute toxicity experiments, 5-MeO-MiPT was added to a blank solution in order to obtain a dose of 0.27 mg/kg in 150 µL injection (n=10) and the last group were injected 2.7 mg/kg 5-MeO-MiPT in a 150 µL injection (n=10). Quantitative toxicological analysis, hystotoxicological and immunohistochemical analysis were performed. RESULTS: In the toxicological analysis, 5-MeO-MiPT was found negative in biological samples which were control, vehicle control, and 0.27 mg/kg dose mice groups. 5-MeO-MiPT was found 2.7-13.4 ng/mL in blood, 11-29 ng/g in kidney, 15.2-108.3 ng/g in liver, and 1.5-40.6 ng/g in the brain in 2,7 mg/kg injected group. In a low dose of the 5-MeO-MiPT liver section, compared with normal tissues, the difference in staining was statistically significant (p<0.0001). In high-dose of 5-MeO-MiPT, H-score showed that the increase in the number of Caspase-3 positive cells was significant compared to the control (p<0.05). In high-dose of 5-MeO-MiPT, intense Caspase-3 immunoreactivity was observed and the increase in the number of Caspase-3 positive cells compared to the control was statistically significant (p<0.05). In brain section, the statistics of the results obtained from the H-score showed that the increase in the number of Caspase-3 positive cells was significant compared to the control (p=0.0183). In vehicle control liver section, there were few Caspase-8 positive cells characterized by a light brown appearance (p=0.0117). In the high-dose 5-MeO-MiPT group, the numbers of positive cells at low and high doses of 5-MeO-MiPT group were statistically significant compared to the control (p<0.05). In the high-dose 5-MeO-MiPT group, Caspase-8 immunoreactivity was detected in the glomerular structures. Compared to control, the increase in Caspase-8 immunoreactivity was found to be statistically significant (p<0.05). CONCLUSION: Low-dose 5-MeO-MiPT did not cause any serious histopathological effects on the liver, kidney, and brain. High doses induce apoptotic cell death through caspase activity.

Morphine Concentrations in Human Urine Following Poppy Seed Paste Consumption.

Papaver somniferum (opium poppy) is one of the world's oldest medicinal plants which are widely used for medicinal, nutritive and scientific purposes. Turkey is one of the major legal opium poppy producer countries in the world and the seed paste of the poppies is consumed in great deal, even more than 100g per meal. The main objective of this study is to investigate the influence of poppy seed paste ingestion on urine tests for opiates whether or not could lead to opiate positive urine test results. For this purpose, a variety of poppies were used and the morphine content of white, yellow and blue-black poppies were determined as 1.9, 4.0 and 2.6mg/kg, respectively. 100g of these seed pastes were consumed in the breakfast by ten healthy adults enrolled in the study over three days and urine samples were collected before and after the breakfast. Opiate screening analysis was carried out by enzyme immunoassay method and the results were evaluated by two different cut-off values (300 and 2000ng/mL). Morphine confirmation analysis was made by GC-MS system and the chromatographic method was validated in terms of selectivity, extraction efficiency, linearity (25-2000ng/ml), intra-assay precision, accuracy, limit of detection (LOD) and limit of quantitation (LOQ) (3 and 10ng/ml), carryover, matrix effect, dilution integrity and stability. According to cut-off value 300ng/ml, opiate concentrations were found positive up to 48h. For cut-off value 2000ng/mL; this time was up to 12h in collected urine samples after consumption of three different colored poppy seed pastes. In all urine samples, thebaine was detected while the heroin abuse metabolite 6-acetyl morphine (6-AM) was not. Urine drug testing legislation was revised on 2016 in Turkey and opiate screening cut-off values increased from 300 to 2000ng/mL. Overall results have shown that poppy seed paste as food consumption could lead to opiate positive urine test result even if increased cut off levels are used. It can also be deduced that thebaine can be taken as supportive biomarker for poppy seed paste consumption. Awareness of interpretation of urine test results and defining the procedures especially for forensic drug testing must be done in legal aspect to ensure justice for each individual (workplace, traffic, court etc.).