Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples.

PURPOSE: Therapeutic drug monitoring of plasma lamotrigine (LTG) has customarily been carried out in order to prevent some its adverse effects. For forensic purposes, determination of LTG in plasma is an useful tool in cases of accidental overdose or suicidal attempts. Currently, there are several analytical methods available including some based on LC tandem mass spectrometry techniques, but simple and accessible LC-UV methods still can be useful for the purpose. Here we report on a new high-performance liquid chromatography method for the determination of lamotrigine in human plasma which has been developed and validated including selectivity, sensitivity, accuracy, precision and recovery studies. METHODS: Lamotrigine and the internal standard chloramphenicol were extracted from plasma using liquid-liquid extraction using small volumes of buffer and ethylacetate. Detection was monitored at 305.7 and 276.0 nm for lamotrigine and chloramphenicol, respectively. RESULTS: The method was linear concentration dependence within the range of 0.1-10 µg/ml, with a mean coefficient of correlation r = 0.993. The limit of detection (LOD) was 0.04 µg/ml and the limit of quantification (LOQ) was 0.1 µg/ml. Intra and interday precision values were lower than 9.0% at all concentrations studied. The intra and interday accuracy values ranged from - 7.6 to 10.1%. Recovery was found to be 98.9% or higher. The method here described was successfully applied to 11 postmortem blood samples received at the Forensic Sciences Institute of Santiago de Compostela (Spain). CONCLUSION: A new HPLC method for the determination of lamotrigine in human plasma was developed and validated. A liquid-liquid extraction using small volumes of buffer and ethylacetate was optimized. The proposed method is suitable for forensic toxicological analysis.

Evaluation of an Oral Fluid Collection Device and a Solid-Phase Extraction Method for the Determination of Coca Leaf Alkaloids by Gas Chromatography-Mass Spectrometry.

Some South American countries have ancient traditions that may pose legal problems, such as the consumption of coca leaves, as this can provide positive results for cocaine use after the analysis of biological samples. For this reason, it is necessary to find specific markers that help differentiate legal from illegal consumption, such as tropacocaine, cinnamoylcocaine, and especially hygrine and cuscohygrine. In this work, two techniques for collecting biological samples are compared: the Quantisal® Oral Fluid collection device and passive drooling. Once the samples were collected, they were subjected to solid-phase extraction for subsequent injection into GC-MS. Different validation parameters included in international guides have been studied to evaluate whether the proposed method is valid for the defined purpose, placing special emphasis on the study of the matrix effect and little value on GC-MS analyses. With respect to this parameter, an increase in the signal was found for CUS and t-CIN, but it was not significant for the rest of the substances studied. The recoveries have varied significantly depending on the way of working, being higher when working with standardized areas. After carrying out work with the oral fluid samples collected from laboratory volunteers, the method was applied to two real samples. The results obtained support the need for further research to overcome certain limitations presented by the device.

Simple Method for the Determination of THC and THC-COOH in Human Postmortem Blood Samples by Gas Chromatography-Mass Spectrometry.

A simple and sensitive analytical method was developed for qualitative and quantitative analysis of Δ9-tetrahydrocannabinol (Δ9-THC) and its metabolite 11-nor-Δ9-tetrahydrocannabinol-carboxylic acid (Δ9-THC-COOH) in human postmortem blood using gas chromatography/mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode. The method involved a liquid-liquid extraction in two steps, one for Δ9-THC and a second one for Δ9-THC-COOH. The first extract was analyzed using Δ9-THC-D3 as internal standard. The second extract was derivatized and analyzed using Δ9-THC-COOH-D3 as internal standard. The method was shown to be very simple, rapid, and sensitive. The method was validated for the two compounds, including linearity (range 0.05-1.5 µg/mL for Δ9-THC and 0.08-1.5 µg/mL for Δ9-THC-COOH), and the main precision parameters. It was linear for both analytes, with quadratic regression of calibration curves always higher than 0.99. The coefficients of variation were less than 15%. Extraction recoveries were superior to 80% for both compounds. The developed method was used to analyze 41 real plasma samples obtained from the Forensic Toxicology Service of the Institute of Forensic Sciences of Santiago de Compostela (Spain) from cases in which the use of cannabis was involved, demonstrating the usefulness of the proposed method.

Statistical Analysis of Toxicological Data of Victims of Traffic Accidents in Galicia (Spain).

Driving under the influence of alcohol or drugs is a very common behavior in our environment and a serious problem for public health. On the one hand, in 2016, 400,000 people died in the world in traffic accidents in which ethanol was involved. On the other hand, traffic accidents in which the use of drugs of abuse other than ethyl alcohol accounted for more than 160,000 deaths worldwide in 2017. The objective of this work is to carry out a review of the 710 cases of people who died in traffic accidents received at the forensic toxicology service of the Institute of Forensic Sciences of the University of Santiago de Compostela (Galicia-Spain) over a period of 10 years (2009-2019). We performed an observational study of period prevalence, in which the following data were collected: age, sex, year, and analytical results in plasma, in the case of being positive. The data collected was subjected to statistical treatment. Of the 710 cases analyzed, 123 correspond to pedestrians and 587 to occupants of vehicles or motorcycles. A total of 77.6% of the deceased were men. At least one psychotropic substance was found in the blood of almost 40% of the victims. The most frequently found substance was ethyl alcohol, which appeared in 231 cases, more frequently in males. The second place is occupied by benzodiazepines, which appeared in 43 cases, followed by cocaine, which was detected in 25 cases. Polydrug use was found in only 44 cases, with the association of ethanol and cocaine being the most commonly found, followed by that of ethanol and benzodiazepines. Only in 5 of the cases analyzed there were 3 or more substances present. With the data obtained in this study, it is shown that in traffic accidents, the finding of different toxic or medicinal substances is frequent. Ethyl alcohol continues to be very present in road accidents (most detected substance), with the great impact that this implies. Secondly, the presence of benzodiazepines stands out, and cocaine is the third most detected toxic in this study. These results allow to obtain a profile of the substances most frequently involved in traffic accidents. Despite the surveillance, control, and information campaigns that the Spanish Government regularly carries out, the results are far from satisfactory.

ANCA-positive vasculitis induced by levamisole-adulterated cocaine and nephrotic syndrome: The kidney as an unusual target.

PATIENT: Male, 36 FINAL DIAGNOSIS: Levamisole-induced vasculopathy Symptoms: Purpuric skin lesions Medication: Levamisole Clinical Procedure: - Specialty: Internal Medicine. OBJECTIVE: Unusual clinical course. BACKGROUND: Levamisole has been detected in seized cocaine samples and a levamisole-induced vasculopathy (LIV) has been described, mainly focused on skin. CASE REPORT: A 36-year-old Caucasian man with history of antibodies to hepatitis C infection (negative hepatitis C virus RNA and negative HIV serology), smoking, and intravenous use of cocaine and brown heroin, presented to the hospital with purpuric skin lesions on extremities and earlobes. One month before the current presentation, a skin punch biopsy of one of these lesions was performed, showing histopathologic findings suggestive of mixed cryoglobulinemia. Laboratory testing revealed leukopenia, renal failure, and nephrotic syndrome. Antimyeloperoxidase antineutrophil cytoplasmic antibodies (MPO-ANCA) were positive. The previous skin punch biopsy was revised and demonstrated pathologic findings consistent with leukocytoclastic vasculitis. An analysis of a cocaine sample for personal use, provided by the patient, was performed using mass spectrometry-gas chromatography and levamisole was detected. Three boluses of intravenous methylprednisolone were administered, followed by oral prednisone 1 mg/Kg per day. Skin lesions and renal function improved. CONCLUSIONS: To our knowledge, this is the first report of nephrotic syndrome induced by levamisole-adulterated cocaine, proven by cocaine sample toxicology. Lack of renal biopsy is a limitation of this report.

Estudio epidemiológico de las intoxicaciones agudas atendidas en un hospital gallego entre 2005 y 2008 / Epidemiological study of acute poisoning cases treated at a Galician hospital between 2005 and 2008

Se realizó un estudio descriptivo retrospectivo de las intoxicaciones agudas registradas en el Complexo Hospitalario de Pontevedra (CHOP) entre enero de 2005 y diciembre de 2008, para conocer el número y tipo de intoxicaciones atendidas, su distribución según el sexo y edad del paciente, cronología, tipo de agentes tóxicos implicados, intencionalidad, antecedentes, clínica, evolución, tratamiento y análisis toxicológico empleado en el diagnóstico. Mediante los programas informáticos Clínica e IANUS y la consulta de todas las historias clínicas en papel de los pacientes atendidos con síntomas de intoxicación, se registraron los datos necesarios. Se incluyeron un total de 1893 pacientes con una edad media de 35,6 ± 17,6 años (66% hombres). La mayor incidencia se alcanzó los sábados y domingos durante los meses de verano (junio, julio y agosto). Los agentes tóxicos más frecuentes fueron las drogas de abuso (70,4%), destacando entre ellas el alcohol etílico (61%), frecuentemente asociado al sexo masculino y a un elevado grado de dependencia. En segundo lugar se situaron las intoxicaciones medicamentosas, frecuentemente asociadas al sexo femenino y producidas, en un 73,2% de los casos, por benzodiacepinas. La mayoría de las intoxicaciones registradas fueron voluntarias y en el 18,8% hubo intencionalidad suicida. Ésta se asoció mayoritariamente al sexo femenino. Las manifestaciones neurológicas fueron las más frecuentes y la mortalidad tan solo fue del 0,2% (AU)

A descriptive retrospective study of acute intoxication cases registered at the Complexo Hospitalario de Pontevedra (CHOP) between January 2005 and December 2008 was performed to find out the number and types of poisoning cases treated, their distribution according to patient’s sex and age, chronology, type of toxic agents involved, intentionality, history, symptoms, clinical development, treatment and toxicological analysis used for diagnosis. Data were recorded using Clinica and IANUS software and consulting all paper records of patients with symptoms of poisoning. Data from a total of 1893 patients with a mean age of 35.6 ± 17.6 years (66% men) were included. Highest rates of poisoning were recorded on Saturdays and Sundays during the summer months (June, July and August). Drugs of abuse were the most common toxic agents (70.4%), ethyl alcohol accounting for 61% of these cases, which often involved males and with people with high degrees of dependency. In second place was poisoning resulting from the abuse of medical drugs, more commonly associated with females, and involving benzodiazepines in 73.2% of cases. The majority of these intoxications were intentional, and suicide attempts accounted for 18.8%. The problems most commonly resulting from the poisoning were neurological, and mortality rate was just 0.2% (AU)

[Epidemiological study of acute poisoning cases treated at a Galician hospital between 2005 and 2008]. / Estudio epidemiológico de las intoxicaciones agudas atendidas en un hospital gallego entre 2005 y 2008.

A descriptive retrospective study of acute intoxication cases registered at the Complexo Hospitalario de Pontevedra (CHOP) between January 2005 and December 2008 was performed to find out the number and types of poisoning cases treated, their distribution according to patient's sex and age, chronology, type of toxic agents involved, intentionality, history, symptoms, clinical development, treatment and toxicological analysis used for diagnosis. Data were recorded using Clinica and IANUS software and consulting all paper records of patients with symptoms of poisoning. Data from a total of 1893 patients with a mean age of 35.6 ± 17.6 years (66% men) were included. Highest rates of poisoning were recorded on Saturdays and Sundays during the summer months (June, July and August). Drugs of abuse were the most common toxic agents (70.4%), ethyl alcohol accounting for 61% of these cases, which often involved males and with people with high degrees of dependency. In second place was poisoning resulting from the abuse of medical drugs, more commonly associated with females, and involving benzodiazepines in 73.2% of cases. The majority of these intoxications were intentional, and suicide attempts accounted for 18.8%. The problems most commonly resulting from the poisoning were neurological, and mortality rate was just 0.2%.

Drogas y teratogenia / Drug of abuse and teratogenicity

Las drogas más usadas en el embarazo siguen siendo el tabaco y el alcohol, dentro de las legales, y el cannabis, seguido de la cocaína dentro de las ilegales. Aunque se ha asociado el consumo de drogas con un amplio abanico de malformaciones estructurales, las drogas de abuso tienen un escaso potencial teratogénico, entendido en el sentido estricto de malformación física, con la excepción del alcohol, que se asocia al síndrome alcohol fetal. Sin embargo, sí se está encontrando asociación entre la exposición prenatal a las drogas de abuso y alteraciones específicas del comportamiento, habitualmente muy sutiles, pero que no deben pasar desapercibidas, y que entran en el concepto actual de teratogénesis. La exposición intraútero a las drogas es un factor de riesgo para la gestación, con frecuencia asociado a otros factores de riesgo concomitantes, como la desnutrición, las enfermedades maternas y la pobreza, y cuyos efectos se solapan, se potencian y se confunden con frecuencia. En este trabajo se abordan de modo resumido los aspectos generales que regulan la teratogénesis (factores que determinan que una determinada exposición pueda ser considerada o no de riesgo, el papel de la placenta, etc), la epidemiología del consumo de drogas durante el embarazo y los factores que dificultan el estudio de estos efectos para estas sustancias. También se hace un análisis de los principales efectos teratogénicos relacionados con las drogas

The problem of the malformed children has concerned the humanity for ever, and this also includes the prenatal exposure to drugs of abuse. Tobacco and alcohol are the most common legal drugs used in pregnancy, and cannabis and cocaine, the most common illegal drugs. The use of drugs has been associated with a wide range of structural malformations, but with the exception of the alcohol and the fetal alcohol syndrome, drugs of abuse seem to have a weak teratogenic potential, in the classical structural malformation conception. Nevertheless, there has been found association between the prenatal exposure to these drugs and specific behavioural alterations, usually very subtle but that should not be ignored. This kind of alterations are now included in the conception of teratogenesis. So, drug use during pregnancy is a risk factor, usually associated to other concomitant risk factors like nutrition deficits, maternal illness and poverty, and these effects are overlapped, potentiated and very often confounded. In this review the main variables involved in the development of teratogenesis (factors that should be take into account in a specific exposition to evaluate the potential risk, the roll of the placenta, etc) are analysed. The epidemiology of the drug abuse in pregnancy and the concomitant factors that make difficult this evaluation are also studied. Finally a review of the main teratogenic effects related to the most common abused drugs was done