Detection of coca alkaloids in oral fluid from coca leaf (tea) consumers: using solid phase extraction to improve validation parameters and widen the detection window.

Hygrine and cuscohygrine, two coca leaf alkaloids, have been previously proposed as markers to differentiate legal and illegal cocaine consumption. This is a very common problem in some countries of South America, where the consumption of coca leaves has a long tradition. Analytical methods focusing on the assessment of coca leaf alkaloids, such as cuscohygrine, hygrine, tropacocaine and t-cinnamoylcocaine, in oral fluid are virtually non-existent in forensic toxicology laboratories worldwide due to their lack of application. However, the problem of differentiating legal and illegal cocaine use in criminal justice, DUID (drug-impaired driving) and WDT (workplace drug testing) programs is growing. Therefore, researchers are obliged to develop methods to measure coca leaf alkaloids (cuscohygrine, hygrine and t-cinnamoylcocaine) in biological matrices for further validation for routine analyses in forensic toxicology laboratories. This work aims to optimize a previously published separation method by protein precipitation in oral fluid by using solid-phase extraction (SPE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) operating in multiple reaction monitoring (MRM) mode. The use of SPE allowed the matrix effect and the background to be reduced in the chromatograms due to the obtained cleaner extracts. Consequently, improved detection and quantification limits were reached. Findings showed that the detection windows for coca leaf alkaloids were longer than three hours in real oral fluid samples from volunteers who drank a cup of coca tea. These detection windows are quite higher than those previously obtained when using the method based on separation by protein precipitation.

Disappearance of codeine, morphine and 6-MAM in hair after cessation of abuse.

Research on the determination of drugs of abuse in hair has established that drugs can be detected in hair even long after cessation of use. The purpose of this study was to analyze hair samples from chronic opioid users who were beginning a controlled drug cessation program. The study population (n = 15) is involved in a drug rehabilitation program in Santiago de Compostela, Spain. Over a 6-month period, subjects provided hair samples at 2-month intervals, with the first sample collected on the day they began the program. Codeine, morphine, and 6-MAM were analyzed by GC/MS (LOQ = 0.2 ng/mg). Hair tresses were divided into 1 cm segments and analyzed for all analytes 0-1 cm corresponding to the proximal portion to the scalp Following cessation of opioid use, traces of codeine, morphine, and 6-MAM still remained in the newly growing hair segments for a specified period. After 2 months, still 27 % of the users tested positive, and at 4 months, 20 % were positive but only for 6-MAM. However, after 6 months of abstinence, the results were negative for all analytes.

Epidemiología de las intoxicaciones agudas en el Servicio de Urgencias del Hospital Clínico Universitario de Santiago de Compostela / Epidemiology of acute poisonings in the Emergency Service of the Clinic University Hospital of Santiago de Compostela

Introducción y métodos: Las intoxicaciones agudas son un motivo frecuente de consulta urgente. Este estudio descriptivo observacional retrospectivo busca analizar la epidemiología de las intoxicaciones agudas atendidas en el Servicio de Urgencias entre junio y noviembre de 2019. Resultados: Las intoxicaciones agudas constituyeron el 1,2% de las consultas de Urgencias (edad media de 44 años, 64,9% hombres). Las drogas de abuso fueron el grupo más frecuente (sobre todo en el sexo masculino), siendo el etanol el principal agente; seguidas del policonsumo y los medicamentos (especialmente en el sexo femenino), destacando las benzodiacepinas. Las intoxicaciones fueron más habituales los fines de semana y durante la noche. La principal modalidad fue la voluntaria (principalmente en hombres), seguida de la autolítica (sobre todo en mujeres). El 26,5% de los pacientes acudieron en ambulancia. El 53,9% de los pacientes presentaba antecedentes toxicológicos (sobre todo hombres) y el 32,3% antecedentes psiquiátricos (principalmente en mujeres). La clínica más frecuente fue la neurológica. Se realizó tratamiento sintomático en la mayoría de los pacientes, empleándose tratamientos específicos en una pequeña proporción. El 84,4% de los pacientes fueron dados de alta desde Urgencias, necesitando atención de la Unidad de Cuidados Intensivos el 2,0%; los ancianos ingresaron más frecuentemente. Conclusiones: Las intoxicaciones agudas revisten poca gravedad en este medio. Hay diferencias en sus características según sexo y edad, así como en la modalidad en función del tóxico y los antecedentes personales. (AU)

Introduction and methods: Acute poisoning is a common reason for consultation in Emergency Services. This retrospective observational study looks for analysing the epidemiology of acute poisonings in the Emergency Service of the Clinic University Hospital of de Santiago de Compostela between June and November 2019. Results: Acute poisonings were 1,2% of Emergency Service consults (mean age 44 years, 64.9% men). Abuse drugs were the most usual group (specially in male sex), being ethanol the principal agent; followed by polycompsumption and medicines (specially in female sex), highlighting benzodiazepines. Poisonings were more frequent in weekends and during the night. The first modality was voluntary (principally in men), followed by suicidal (principally in women). 26.5% of patients came by ambulance. 53.9% of patients showed toxicological history (specially men) and 32.3% had psychiatric history (specially in women). The most common clinical manifestations were neurological. Most patients got symptomatic treatment, using specific treatment in a small proportion. 84.4% of patients were discharged from the Emergency Service, needing attention from the Intensive Care Unit 2.0%; the elderly patients were admitted more frequently. Conclusions: Acute poisonings have little gravity in this environment. There are differences in their features according to sex and age, as wells as in modality depending on the toxic agent and the personal history. (AU)

Sexual Assault: Approach to Reality in the Area of Santiago de Compostela (Galicia, Spain) through a 12-Year Retrospective Study.

Sexual assault constitutes a severely traumatic experience that impacts the lives of far too many victims each year. The underlying behaviors of the offenders are often associated with psychological, physical and social distress, and the use of psychotropic substances was found in a good number of cases. A descriptive and retrospective review of sexual assault cases has been undertaken to identify trends in the toxicology findings in Drug-Facilitated Sexual Assault in Santiago de Compostela over the past 12 years. During this period, a total of 69 cases were referred to the Forensic Toxicology Service as sexual assault cases. The sex and age distribution of the cases showed that females between the ages of 14 and 65 years constituted the group most frequently submitted to sexual assault, with a peak of 55.1% in the 18- to 30-year age group. Alcohol consumption was positive in 77.1% of positive cases determined, followed by drugs (34.4%) and illicit drugs (26.2%). Our results showed a high percentage of alcohol consumption in sexual assault cases.

Epidemiología de las intoxicaciones agudas en el Servicio de Urgencias del Hospital Clínico Universitario de Santiago de Compostela / Epidemiology of acute poisonings in the Emergency Service of the Clinic University Hospital of Santiago de Compostela

Introducción y métodos. Las intoxicaciones agudas son unmotivo frecuente de consulta urgente. Este estudio descriptivoobservacional retrospectivo busca analizar la epidemiología de lasintoxicaciones agudas atendidas en el Servicio de Urgencias entre junioy noviembre de 2019. Resultados. Las intoxicaciones agudasconstituyeron el 1,2% de las consultas de Urgencias (edad media de 44años, 64,9% hombres). Las drogas de abuso fueron el grupo másfrecuente (sobre todo en el sexo masculino), siendo el etanol elprincipal agente; seguidas del policonsumo y los medicamentos(especialmente en el sexo femenino), destacando las benzodiacepinas.Las intoxicaciones fueron más habituales los fines de semana y durantela noche. La principal modalidad fue la voluntaria (principalmente enhombres), seguida de la autolítica (sobre todo en mujeres). El 26,5%de los pacientes acudieron en ambulancia. El 53,9% de los pacientespresentaba antecedentes toxicológicos (sobre todo hombres) y el 32,3%antecedentes psiquiátricos (principalmente en mujeres). La clínica másfrecuente fue la neurológica. Se realizó tratamiento sintomático en lamayoría de los pacientes, empleándose tratamientos específicos en unapequeña proporción. El 84,4% de los pacientes fueron dados de altadesde Urgencias, necesitando atención de la Unidad de CuidadosIntensivos el 2,0%; los ancianos ingresaron más frecuentemente.Conclusiones. Las intoxicaciones agudas revisten poca gravedad eneste medio. Hay diferencias en sus características según sexo y edad,así como en la modalidad en función del tóxico y los antecedentespersonales. (AU)

Introduction and methods. Acute poisoning is a common reason for consultation in Emergency Services. This retrospective observational study looks for analysing the epidemiology of acute poisonings in the Emergency Service of the Clinic University Hospital of de Santiago de Compostela between June and November 2019. Results. Acute poisonings were 1,2% of Emergency Service consults (mean age 44 years, 64.9% men). Abuse drugs were the most usual group (specially in male sex), being ethanol the principal agent; followed by polycompsumption and medicines (specially in female sex), highlighting benzodiazepines. Poisonings were more frequent in weekends and during the night. The first modality was voluntary (principally in men), followed by suicidal (principally in women). 26.5% of patients came by ambulance. 53.9% of patients showed toxicological history (specially men) and 32.3% had psychiatric history (specially in women). The most common clinical manifestations were neurological. Most patients got symptomatic treatment, using specific treatment in a small proportion. 84.4% of patients were discharged from the Emergency Service, needing attention from the Intensive Care Unit 2.0%; the elderly patients were admitted more frequently. Conclusions. Acute poisonings have little gravity in this environment. There are differences in their features according to sex and age, as wells as in modality depending on the toxic agent and the personal history. (AU)

Determination of Seven Antidepressants in Pericardial Fluid by Means of Dispersive Liquid-Liquid Microextraction (DLLME) and Gas Chromatography-Mass Spectrometry (GC/MS).

Although blood is often used to detect and quantify the presence of drugs, there are some instances where samples obtained from other biological matrices, like pericardial fluid (PF), are necessary since adequate blood samples may not be available. PF is an epicardial transudate, which contains plasma components that include toxicological substances making this sample useful when blood samples are not available. This fluid is a well preserved postmortem sample and can easily be collected in larger amounts without significant contamination, compared with other body fluids. Although studies involving PF began around the 1980's, the adequacy of such fluid as a biological matrix has been poorly investigated. Antidepressants are frequently detected in postmortem samples from forensic cases. Nowadays, they constitute some of the most commonly prescribed drugs worldwide. A total of seven antidepressants (venlafaxine, mirtazapine, olanzapine, paroxetine, sertraline, fluoxetine and citalopram) were evaluated in this study. A new extraction method involving dispersive liquid-liquid microextraction (DLLME) is presented in which chloroform and acetonitrile are determined to be the best extraction and dispersing solvents. The experimental design was achieved using StatGraphics 18. The Response Surface Methodology enabled us to know the optimal volume for the two solvents used in the DLLME. The detection technique used was gas chromatography-mass spectrometry (GC-MS) with electron impact ionization as ionization source. A temperature gradient has been used and the total chromatographic separation time was 19.43 min. Validation results met the international validation guidances (FDA). Under the optimal condition, the method offered good validation parameters showing a new efficient, simple, rapid, and sensitive method. The analytical method was applied to thirty-one pericardial fluid samples. Twenty-one samples were positive with concentrations between 0.19 and 8.48 µg/mL. Venlafaxine and olanzapine were the antidepressants most frequently found.

Determination of benzodiazepines in pericardial fluid by gas chromatography-mass spectrometry.

In Forensic Toxicology it is sometimes impossible to obtain a valid blood sample to perform toxicological analysis due to several factors like advanced state of decomposition, severe burns, bleed to death…. Pericardial Fluid has already been studied during the last years and has been proposed as a valid specimen for toxicological tests. Over the years, the consumption of benzodiazepines spread among the drug dependent population and became noticeable in drug facilitated assault cases and road accidents. Improvement of the analytical methodology required for detecting the presence of these drugs in biological samples is of great importance for forensic toxicology, in order to correctly diagnose an exposure or a poisoning. In this study, 9 benzodiazepines (diazepam, nordiazepam, midazolam, bromazepam, oxazepam, temazepam, lorazepam, clonazepam and alprazolam) have been determined in pericardial fluid. For this purpose a solid phase extraction (SPE) was carried out using Bond Elut Certify cartridges. After the derivatization of six of the nine benzodiazepines, gas chromatography coupled to a selective mass detector was used as the technique for the separation of the analytes. The method developed was fully validated for the 9 analytes and was applied to real samples of pericardial fluid received at the Forensic Toxicology Service of the University of Santiago de Compostela. Finally, they were compared with blood results looking for the existence of a possible correlation between both biological samples.

Deaths from acute drug reactions in Galician (Spain) Prisons (2001-2010).

INTRODUCTION AND OBJECTIVES: drug use is associated with multiple complications with an increase in morbidity, with death by acute drugs reactions (ADR) being the most serious. A large percentage of the prison population has problems associated with drug additions, and substance abuse is also a common internal problem of penal institutions, despite their control measures. The goal of this study is to analyse the prevalence of ADR in penitentiaries, deceased sociodemographic characteristics as well as the circumstances in which they are produced. MATERIAL AND METHODS: All deaths by ADR between 2001-2010 in Galicia are studied, in particular, those deaths that took place inside prisons. RESULTS: In the whole sample (n=510) male (90.6%), single (46.1%) with an average age of 35.8 and with a prevalent factor of long experience in drug abuse. Thirty seven of them died in Penal/Correctional Institutions, representing 7.3% of the total sample. The characteristics of this population subtype were similar to the total sample (average age: 34.7 years; 89.2% were males) but we found significant differences regarding the substances detected. DISCUSSION: ADR is the most frequent cause of death among drug addict convicts in prisons. The pattern of the detected substances in the toxicological analysis as well as the socio-demographic characteristics can help to establish a higher risk profile and preventive measures.

Chromium available fractions in arousa sediments using a modified microwave BCR protocol based on microwave assisted extraction.

In a research of chromium availability, the three-stage sequential extraction procedure, proposed by European Community Bureau of Reference (BCR), has been applied for the metal fractionation in marine sediment samples. The procedure has been modified, evaluating the effect of microwave energy to perform the sequential method. Results achieved a substantial reduction in time in comparison to the traditional shaking technique. The time of the first and the second extractions were reduced to 30s in both steps using microwave heating and working at 66W of power. To the third extraction, higher power was necessary so it was chosen to work at 198W. In the last step investigated, time was reduced to 1min, being a great improvement respect to the conventional BCR sequential extraction protocol. Chromium determinations in these extracts were carried out by Electrothermal Atomic Absorption Spectrometry (ETAAS). The developed method was applied for chromium determination in marine sediment samples from Ria de Arousa (Galicia, NW of Spain). The values obtained by our laboratory ranged from 0.10 to 1.02mugg(-1) for soluble and reducible fractions, and from 5.5 to 60.0mugg(-1) for the oxidisable fraction. The higher concentrations obtained were the chromium associated with the organic fraction of the marine sediments.

Silicon determination in milk by electrothermal atomic absorption spectrometry using palladium as chemical modifier.

A direct method for silicon determination in milk samples by Electrothermal Atomic Absorption Spectrometry was developed. Palladium was used as chemical modifier at a concentration of 610 mg L(-1); with this modifier, silicon was stable up to 1800 degrees C. The precision and accuracy of the method were investigated. The detection limit was 16.2, 2.7 and 7.2 micro g L(-1) for cows' milk, human milk and infant formula, respectively. The method was applied to silicon determination in 17 infant formula samples, 13 human milk samples and 12 cows' milk samples.

Selective preconcentration and determination of tributyltin in fresh water by electrothermal atomic absorption spectrometry.

A rapid and simple method for separation and determination of tributyltin (TBT) in mineral and tap water is described. The procedure is based on the selective retention of TBT by a chelating resin, Amberlite XAD-2 impregnated with tropolone. The addition of 0.8% sulfuric acid to the water sample leads to the retention of TBT by the resin while monobutyltin (MBT), dibutyltin (DBT) and inorganic tin remain in solution. TBT is eluted with methyl isobutyl ketone (MIBK) obtaining a preconcentration factor of 80. Tin concentration is determined by ETAAS using zirconium coated tubes. Multi-injection and hot injection techniques are used in order to enhance the sensitivity of the method. A detection limit of 14.4 ng L(-1) is achieved with recoveries near to 100%. The procedure has been successfully applied to TBT determination in various fresh water samples.

Separation of gallium and indium from ores matrix by sorption on Amberlite XAD-2 coated with PAN.

A chelating sorbent obtained by adsorption of 1-(2-pyridylazo)-2-naphthol (PAN) on Amberlite XAD-2 was used for the preconcentration of Ga and In. The analytical characteristics of the chelating sorbent were investigated and optimun sorption conditions for these metals under dynamic conditions were established. A peristaltic pump is used to adjust the flow rate of the solution. Elements are collected from the column by using a mixture adjusted to a pH range of 4-7 and 6-12 by ammonia or ammonium chloride for Ga and In, respectively. The procedure developed was applied to the analysis of different ores.

Experimental designs in the optimisation of ultrasonic bath-acid-leaching procedures for the determination of trace elements in human hair samples by atomic absorption spectrometry.

Experimental designs were used for the optimisation of acid-leaching procedures assisted by ultrasonic energy for the extraction of Ca, Cu, Fe, Mg, Mn and Zn from human hair samples. A Plackett-Burman 2(7) x 3/32 design for seven factors ([HNO3], [HCl], [H2O2], acid/oxidant solution volume, exposure time to ultrasounds, temperature of the ultrasonic bath and hair particle size) was used in order to choose the variables affecting the acid-leaching process. The variables [HNO3], [HCl] and temperature of the ultrasonic bath were found to be the most important parameters for the acid-leaching procedure, and these variables were optimised by a response surface design (central composite design 2(3) + star) which involved 16 experiments. Optimum values in the 3.7-4.2 M range were found for [HNO3], while optimum values between 3.0 and 3.5 M were found for [HCl]. The optimum temperature of the ultrasonic bath was between 80 and 90 degrees C. An acid digestion induced by microwave energy (details given) was used to obtain the total metal concentration and also for comparative purposes. Ca, Cu, Fe, Mg and Zn were measured by flame atomic absorption spectrometry (FAAS) using a conventional air/acetylene flame, while Mn was determined by electrothermal atomic absorption spectrometry (ETAAS) under optimised conditions. Two different reference materials, IAEA-085 International Atomic Energy Agency, Monaco) and NIES No. 13 (National Institute for Environmental Studies, Japan), with certified metal contents for some of the elements investigated, were used in order to verify the accuracy of the methods.

Selenium determination in mother and child's hair by electrothermal atomic absorption spectrometry.

A method for the selenium determination in a mother and her child's hair using palladium as a chemical modifier was optimized. The sample was digested with nitric acid and hydrogen peroxide and diluted to 5 ml. To achieve complete mineralization the samples were ashed at 1200 degrees C in the presence of palladium as a chemical modifier. The optimum atomization temperature was 1900 degrees C. The precision and accuracy of the method were studied using the reference material CRM 397. Results of calibration using aqueous standards and the standard addition method were compared. The method was applied to the selenium determination in 30 samples of the mother's and child's hair. The levels found were 0.54 +/- 0.34 microgram/g for mother's hair and 0.77 +/- 0.25 microgram/g for child's hair.

A study of illicit cocaine seizure classification by pattern recognition techniques applied to metal data.

Fifteen metallic species, silver (Ag), aluminum (Al), calcium (Ca), cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), sodium (Na), nickel (Ni), lead (Pb), strontium (Sr) and zinc (Zn), were determined in 46 cocaine samples confiscated by the Spanish police in Galicia (northwest Spain). Classification of these cocaine samples according to their geographic origin (Colombia and Venezuela) was achieved by the application of pattern recognition techniques to the metallic content data. Cocaine samples, around 0.5 g, were directly dissolved in 2 mL of 35.0% (v/v) HNO3, diluted to 10 mL with ultrapure water. The metals were quantified by means of electrothermal atomic absorption spectrometry (Ag, Al, Cd, Cr, Cu, Mn, Ni, Pb and Sr), flame atomic absorption spectrometry (Ca, Fe, Mg and Zn), and flame atomic emission spectrometry (K and Na). Results show that two geographic origins can be established through the presence of trace and major elements.

Direct electrothermal atomic absorption spectrometry determination of nickel in sea water using multiple hot injection and Zeeman correction.

Methods for the direct determination of Ni in sea water samples by ETAAS were developed using Zeeman effect background correction system (ZEBC) and a multi-injection technique. A mass of palladium nitrate of 2.5 mug (for an injection volume of 100 mul) was used as chemical modifier. The optimum pyrolysis and atomization temperatures were 1700 and 2100 degrees C, respectively. The characteristic mass (m(0)) and characteristic concentration (C(0)), precision and accuracy were studied for different injection volumes (20, 100 and 200 mul). For an injection volume of 100 mul (five 20 mul aliquot) of sample the accuracy analysis of different certified materials (saline and non saline water) was agreeable. The total time of the proposed procedure is 6 min. A m(0) and C(0) of 34.5 pg and 0.3 mug l(-1), respectively were obtained for this injection volume (100 mul). Finally, interferences from major and minor components of sea water was studied.

Determination of aluminium and manganese in human scalp hair by electrothermal atomic absorption spectrometry using slurry sampling.

Methods for the determination of aluminium and manganese in human scalp hair samples by electrothermal atomic absorption spectrometry using the slurry sampling technique were developed. Palladium and magnesium nitrate were used as chemical modifiers. Hair samples were pulverized using a zirconia vibrational mill ball, and were prepared as aqueous slurries. Determinations can be performed in the linear ranges of 1.9-150 mug l(-1) Al(3+) and 0.03-10.0 mug l(-1) Mn(2+). Limits of detection of 0.9 mg kg(-1) and 27.6 mug kg(-1) were obtained for aluminium and manganese, respectively. The analytical recoveries were between 99.6 and 101.8% for aluminium and in the 98.3-101.3% range for manganese. The repeatability of the methods (n=11), slurry preparation procedure and ETAAS measurement, was 16.0 and 7.9% for aluminium and manganese, respectively. The methods were finally applied to the aluminium and manganese determination in 25 scalp hair samples from healthy adults. The levels for aluminium were between 8.21 and 74.08 mg kg(-1), while concentrations between 0.03 and 1.20 mg kg(-1) were found for manganese.

Column preconcentration of organotin with tropolone-immobilized and their determination by electrothermal atomization absorption spectrometry.

A method for the determination of total organic tin from marine water samples by electrothermal atomization absorption spectrometry (ETAAS) is described. Samples are previously preconcentrated with a chelating molecule (tropolone) impregnated on a macroporous polymer (Amberlite XAD-2). The graphite furnace programme and preconcentration parameters were optimized. Calibration and addition graphs were performed. Sensitivity obtained with this procedure was 13 ng l(-1). Relative standard deviation was always >10% and analytical recovery were satisfactory, approximately 100%. Some possible interferences were investigated, having no problems with this factor. This procedure allows the distinction between organotin compounds and inorganic tin IV, since the latter is not retained on the column.

Effectiveness of palladium as a chemical modifier for direct silver and manganese determination in cocaine and heroin by electrothermal atomic absorption spectrometry.

Methods for the direct determination of silver and manganese in cocaine and heroin by electrothermal atomic absorption spectrometry using palladium as chemical modifier have been developed. Cocaine samples, 0.5 g, were dissolved in 2 ml of 35.0% (m/v), HNO(3) diluting to 10 ml with ultrapure water. A comparative study of palladium, magnesium nitrate, palladium-magnesium nitrate and nitric acid used as chemical modifiers was carried out, and it was found that palladium alone is adequate to stabilize silver and manganese at charring temperatures of 1000 and 1300 degrees C respectively. A study was made on the insertion of a cooling step before the atomization step, and this approach was not advantageous. Limits of detection of 2.3 and 6.9 microg kg(-1) were obtained for silver and manganese respectively. The analytical recovery oscillated between 95.9 and 103.6% for silver and between 99.7 and 103.0% for manganese. Finally, a study of some interferences and a study of the precision and analytical recovery of the amount of sample were also carried out.