[1,4-diaminobenzene and diphenylamine-induced severe methemoglobinaemia treated by hyperbaric oxygen therapy - Case report]. / Zastosowanie tlenoterapii hiperbarycznej w ciezkiej methemoglobinemii wywolanej zatruciem 1,4-diaminobenzenem i difenyloamina ­ opis przypadku.

Methemoglobin is an oxidized form of hemoglobin that is not capable of carrying oxygen. Exposure to exogenous oxidizing agents can cause severe methemoglobinemia and subsequent hypoxia. This article presents the rare case of a 23-year old patient with methemoglobinemia of 54.2% of total hemoglobin, induced by workplace exposure to 1,4-diaminobenzene and diphenylamine. Following the hyperbaric oxygen therapy the patient was discharged in good general condition and referred to a district hospital for observation. Med Pr 2018;69(3):345-350.

Hemorrhagic stroke caused by xylometazoline poisoning.

An increase in the use of over-the-counter medications has been observed in recent years. This also concerns xylometazoline, approved for the treatment of allergic rhinitis or upper respiratory tract infections. We present the fatal case of a 40 year-old-woman with a massive hemorrhagic stroke. Initial toxicology tests of biological material collected during autopsy revealed the presence of xylometazoline. No other significant toxicology findings were noted. LC-MS/MS method has been developed to determine xylometazoline concentration, which was 18.6 ng/mL in blood and 498.9 ng/mL in urine. The macroscopically detected hemorrhagic focus was confirmed by histopathological which confirmed hemorrhagic infarcts in the brain tissue, especially in the subarachnoid area. No other pathological changes were found. Based on findings from autopsy and toxicological analyses, the direct cause of death was concluded to be hemorrhagic stroke resulting from xylometazoline intoxication. Although xylometazoline products are regarded as relatively safe and are available over the counter, the risk of adverse effects, in particular stroke leading to death, should be considered. If adverse effects are observed, it is reasonable to measure the concentration of the drug in blood and urine. With such data, it will be possible to assess the actual exposure to this xenobiotic and draw firmer conclusions.

Histamine and Serotonin Levels in Bone Marrow Stem Cells Niche as Potential Biomarkers of Systemic Mastocytosis and Myeloproliferative Disorders.

Bone marrow studies currently provide a lot of valuable information in the diagnostics of hematological diseases including hematopoietic stem cells disorders. Our studies on low-molecular weight organic compounds in bone marrow stem cell niche in various pathogenic conditions, revealed relatively high variability of histamine levels in different groups of hematological diseases. It was also found that serotonin levels were significantly lower than those typically measured in peripheral blood as well as many have the influence on stem cells proliferative potential. This paper presents findings from quantitative and statistical analyses of histamine and serotonin levels. Bone marrow collected from patients undergoing routine diagnostic procedures for hematological diseases and receiving inpatient treatment were analyzed. Histamine and serotonin levels were measured using hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry. Obtained data were analyzed statistically and correlated with the diagnosed groups of hematological diseases and the parameters of complete blood counts. Histamine was found in all tested samples, including those from patients without malignancy, and the reported levels were comparable to the reference values in blood. This observation allows us to assume that bone marrow cells can produce and accumulate histamine. Moreover, the statistical analysis revealed a significant relationship between histamine levels and diagnosed mastocytosis, and between histamine levels and myeloproliferative neoplasms. Different results were obtained for serotonin, and its concentrations in most cases were below the limit of quantification of the method used (< 0.2 ng/mL), which can only be compared to peripheral blood plasma. In a few cases, significantly higher serotonin levels were observed and it concerned diseases associated with an increased number of megakaryocytes in the bone marrow.

Pharmacokinetics of Levodopa and 3-O-Methyldopa in Parkinsonian Patients Treated with Levodopa and Ropinirole and in Patients with Motor Complications.

Parkinson's disease (PD) is a progressive, neurodegenerative disorder primarily affecting dopaminergic neuronal systems, with impaired motor function as a consequence. The most effective treatment for PD remains the administration of oral levodopa (LD). Long-term LD treatment is frequently associated with motor fluctuations and dyskinesias, which exert a serious impact on a patient's quality of life. The aim of our study was to determine the pharmacokinetics of LD: used as monotherapy or in combination with ropinirole, in patients with advanced PD. Furthermore, an effect of ropinirole on the pharmacokinetics of 3-OMD (a major LD metabolite) was assessed. We also investigated the correlation between the pharmacokinetic parameters of LD and 3-OMD and the occurrence of motor complications. Twenty-seven patients with idiopathic PD participated in the study. Thirteen patients received both LD and ropinirole, and fourteen administered LD monotherapy. Among 27 patients, twelve experienced fluctuations and/or dyskinesias, whereas fifteen were free of motor complications. Inter- and intra-individual variation in the LD and 3-OMD concentrations were observed. There were no significant differences in the LD and 3-OMD concentrations between the patients treated with a combined therapy of LD and ropinirole, and LD monotherapy. There were no significant differences in the LD concentrations in patients with and without motor complications; however, plasma 3-OMD levels were significantly higher in patients with motor complications. A linear one-compartment pharmacokinetic model with the first-order absorption was adopted for LD and 3-OMD. Only mean exit (residence) time for 3-OMD was significantly shorter in patients treated with ropinirole. Lag time, V/F, CL/F and tmax of LD had significantly lower values in patients with motor complications. On the other hand, AUC were significantly higher in these patients, both for LD and 3-OMD. 3-OMD Cmax was significantly higher in patients with motor complications as well. Our results showed that ropinirole does not influence LD or 3-OMD concentrations. Higher 3-OMD levels play a role in inducing motor complications during long-term levodopa therapy.

Selected Hematological Biomarkers to Predict Acute Mortality in Emergency Department Patients. Recent Polish Hospital Statistics.

BACKGROUND: Complete blood count (CBC), red cell distribution width (RDW), mean platelet volume (MPV), mean corpuscular volume (MCV), mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), or platelet (PLT) count are referred as predictors of adverse clinical outcomes in patients. The aim of the research was to identify potential factors of acute mortality in Polish emergency department (ED) patients by using selected hematological biomarkers and routine statistical tools. METHODS: The study presents statistical results on patients who were recently discharged from inpatient facilities within one month prior to the index ED visit. In total, the analysis comprised 14,881 patients with the first RDW, MPV, MCV, MCH, MCHC, or PLT biomarkers' measurements recorded in the emergency department within the years 2016-2019 with a subsequent one month of all-cause mortality observation. The patients were classified with the codes of the International Statistical Classification of Diseases and Related Health Problems after 10th Revision (ICD10). RESULTS: Based on the analysis of RDW, MPV, MCV, MCH, MCHC, and PLT on acute deaths in patients, we establish strong linear and quadratic relationships between the risk factors under study and the clinical response (P < 0.05), however, with different mortality courses and threats. In our statistical analysis, (1) gradient linear relationships were found for RDW and MPV along an entire range of the analyzed biomarkers' measurements, (2) following the quadratic modeling, an increasing risk of death above 95 fL was determined for MCV, and (3) no relation to excess death in ED patients was calculated for MCH, MCHC, and PLT. CONCLUSION: The study shows that there are likely relationships between blood counts and expected patient mortality at some time interval from measurements. Up to 1 month of observation since the first measurement of an hematological biomarker, RDW and MPV stand for a strong relationship with acute mortality of patients, whereas MCV, MCH, MCHC, and PLT give the U-shaped association, RDW and MPV can be established as the stronger predictors of early deaths of patients, MCV only in the highest levels (>95 fL), whereas MCH, MCHC, and PLT have no impact on the excess acute mortality in ED patients.

Red Blood Cells Mean Corpuscular Volume (MCV) and Red Blood Distribution Width (RDW) Parameters as Potential Indicators of Regenerative Potential in Older Patients and Predictors of Acute Mortality - Preliminary Report.

This study presents the statistical results of patients who had been recently discharged from hospital within one month after their treatment in the emergency department (ED). Using routine (14,881) MCV and RDW measurements and statistical tools, we could predict acute mortality in these patients (N = 1158), adjusted for age. It is likely that an increase in the MCV and RDW parameters may correlate in some of our older patients with a poor prognosis with an increased level of circulating IGF-I, which affects red blood cell parameters. The research presents the prognostic statistics of the analyzed clinical factors as well as speculates on the potential correlation of these parameters with the regenerative potential of stem-cell compartment. Analysis shows that both MCV and RDW are statistically significant (Area Under Curve [AUC], lower CI 95% >50%) predictors of acute mortality in ED patients. The classification of patients based on their MCV threshold (= 92.2 units) indicates a proper clinical prognosis in nearly 6 of 10 subjects (AUC >58%), whereas taking into account RDW (=13.8%) indicates a proper clinical prognosis in no more than 7 of 10 individuals. The report concludes that by employing strongly fitting (95%) quadratic modeling of the ORs against the biomarkers studied, one can notice a similar relationship between MCV and RDW as diagnostic tools to predict regenerative potential and clinical outcomes in older patients. Although RDW alone had a 10% higher diagnostic value in terms of predicting early death in the emergency department in patients that were admitted to the ED and subsequently hospitalized, also taking the MCV measurement improved accuracy in predicting clinical outcomes by 2.5% compared to RDW alone.

Toxicological pitfalls in ICU practice.

Either analgosedation or central nervous system dysfunction may be a side effect of implemented pharmacological treatment, as well as a consequence of intentional or unintentional poisoning. In traumatic lesions or anoxia of the central nervous system, a question arises after a recommended follow-up period about the effects of xenobiotics on nervous system function. Although therapeutic drug monitoring is the gold standard in such cases, usually a single toxicological estimation of "a neurodepressive compound" is performed after treatment discontinuation in order to determine the type and amount of exogenous substances, or their metabolites, in a patient's bodily fluids, which allows for an assessment of its actual effects on central nervous system functions. The aim of this paper was to describe the aspects of diagnostic toxicology which are essential for improved determination of the type and amount of exogenous substances present in biological fluids of intensive care patients. We present examples of clinical cases in order to discuss the most common discrepancies in interpretation related to the ordering of toxicology tests.

Daily Onset of Light and Darkness Differentially Controls Hematopoietic Stem Cell Differentiation and Maintenance.

Hematopoietic stem and progenitor cells (HSPCs) tightly couple maintenance of the bone marrow (BM) reservoir, including undifferentiated long-term repopulating hematopoietic stem cells (LT-HSCs), with intensive daily production of mature leukocytes and blood replenishment. We found two daily peaks of BM HSPC activity that are initiated by onset of light and darkness providing this coupling. Both peaks follow transient elevation of BM norepinephrine and TNF secretion, which temporarily increase HSPC reactive oxygen species (ROS) levels. Light-induced norepinephrine and TNF secretion augments HSPC differentiation and increases vascular permeability to replenish the blood. In contrast, darkness-induced TNF increases melatonin secretion to drive renewal of HSPCs and LT-HSC potential through modulating surface CD150 and c-Kit expression, increasing COX-2/αSMA+ macrophages, diminishing vascular permeability, and reducing HSPC ROS levels. These findings reveal that light- and darkness-induced daily bursts of norepinephrine, TNF, and melatonin within the BM are essential for synchronized mature blood cell production and HSPC pool repopulation.

Fatal Intoxication with α-PVP, a Synthetic Cathinone Derivative.

This study presents the fatal case of a young man who was admitted to the ICAU due to sudden cardiac arrest. An interview revealed that the patient had taken some unspecified crystals. From the moment of admission, his condition deteriorated dramatically as a result of increasing circulatory insufficiency. After a few hours, sudden cardiac arrest occurred again and the patient was pronounced dead. In the course of a medicolegal autopsy, samples of biological material were preserved for toxicology tests and histopathological examination. The analysis of samples using the LC-MS/MS technique revealed the presence of α-PVP in the following concentrations: blood-174 ng/mL, urine-401 ng/mL, brain-292 ng/g, liver-190 ng/g, kidney-122 ng/g, gastric contents-606 ng/g. The study also presents findings from the parallel histopathological examination. Based on these findings, cardiac arrest secondary to intoxication with alpha-PVP was determined as the direct cause of the patient's death.

[Amanitine determination as an example of peptide analysis in the biological samples with HPLC-MS technique].

Introduction: Routine toxicological analysis is mostly focused on the identification of non-organic and organic, chemically different compounds, but generally with low mass, usually not greater than 500­600 Da. Peptide compounds with atomic mass higher than 900 Da are a specific analytical group. Several dozen of them are highly-toxic substances well known in toxicological practice, for example mushroom toxin and animal venoms. In the paper the authors present an example of alpha-amanitin to explain the analytical problems and different original solutions in identifying peptides in urine samples with the use of the universal LC MS/MS procedure. Materials and methods: The analyzed material was urine samples collected from patients with potential mushroom intoxication, routinely diagnosed for amanitin determination. Ultra filtration with centrifuge filter tubes (limited mass cutoff 3 kDa) was used. Filtrate fluid was directly injected on the chromatographic column and analyzed with a mass detector (MS/MS). Results: The separation of peptides as organic, amphoteric compounds from biological material with the use of the SPE technique is well known but requires dedicated, specific columns. The presented paper proved that with the fast and simple ultra filtration technique amanitin can be effectively isolated from urine, and the procedure offers satisfactory sensitivity of detection and eliminates the influence of the biological matrix on analytical results. Another problem which had to be solved was the non-characteristic fragmentation of peptides in the MS/MS procedure providing non-selective chromatograms. It is possible to use higher collision energies in the analytical procedure, which results in more characteristic mass spectres, although it offers lower sensitivity. Conclusions: The ultra filtration technique as a procedure of sample preparation is effective for the isolation of amanitin from the biological matrix. The monitoring of selected mass corresponding to transition with the loss of water molecule offers satisfactory sensitivity of determination.

H nuclear magnetic resonance spectroscopic investigation of urine for diagnostic and clinical assessment of methanol intoxication.

Occasional or suicidal methanol intoxications are a permanent problem for most Poisoning Centers around the World. Therefore it is important to look for new diagnostic and clinical prognostic methods. In the present paper 5 cases of methanol intoxication were analyzed. At first the methanol concentrations in blood and urine were estimated with headspace gas chromatography technique. Next the urine samples were examined with 1H NMR spectroscopy, then the levels of ethanol, methanol and its metabolite, formate, lactate and trimethylamine-N-oxide with dimethylamine were evaluated. The concentrations of the above compounds were correlated with the patient's clinical status, the level of ethanol and methanol and biochemical parameters. The results indicate the correlation between clinical course of intoxication, prognostication and lactate level. There were no significant parallels for formate level as acidosis causing metabolite and initial methanol levels. In the urine samples of intoxicated patients the increased trimethyl-N-oxide and dimethylamine levels were observed, which may indicate renal cortex damage. Contrary to the opinion of some clinicians, methanol intoxication may be connected with renal functional disturbances. 1H NMR examination of urine appears to be an excellent tool to evaluate the clinical course of methanol intoxication.

Elaidic acid and vaccenic acid in the plasma of pregnant women and umbilical blood plasma.

INTRODUCTION: The studies suggest that trans fatty acids are the most harmful type of fatty acids, and their ingestion leads to an increase in oxidative stress, changes in the plasma levels of LDL cholesterol, and a reduction in n-6 and n-3 fatty acids utilization. The objective of this study was to determine the concentration profiles of elaidic and vaccenic acids in maternal blood plasma in the perinatal period, and in the plasma of blood collected from the umbilical vein. The article deals with the issue of the correlation between the levels of trans-unsaturated fatty acids in maternal and neonatal blood, which could suggest the transplacental transport of these acids. The potential correlations between the levels of trans-unsaturated fatty acids in maternal and umbilical blood and birth weight and body length in neonates was also verified. MATERIAL AND METHODS: The study group comprised 53 pregnant patients of the Clinic of Obstetrics and Perinatology of the Pomeranian Medical University in Szczecin, aged 18-39 years, together with their newborn babies. Elaidic and vaccenic acid levels were determined in maternal and umbilical plasma. Fatty acid methyl esters were injected onto the capillary column of the Agilent 6890M gas chromatography system integrated with an autosampler. The content of individual fatty acids was determined from the reference curves and expressed in mg/mL. RESULTS: The lowest levels of both acids were observed in umbilical blood plasma. The level of elaidic acid was 0.0019 mg/mL ± 0.0013 mg/mL, while the level of vaccenic acid was 0.0007 mg/mL ± 0.0011 mg/mL. A statistically significant negative correlation was observed between maternal vaccenic acid levels and umbilical vaccenic acid levels. CONCLUSIONS: The presence of trans-unsaturated fatty acids (elaidic acid and vaccenic acid) in umbilical plasma is evidence of the transplacental transport of these acids. The significantly higher levels of elaidic acid compared to vaccenic acid in umbilical plasma suggests that vaccenic acid is more extensively metabolized in placenta. The levels of trans-unsaturated fatty acids had no effect on birth weight and body length in neonates.

[Trans fatty acids (elaidic and vaccenic) in the human milk]. / Wystepowanie nienasyconych kwasów tluszczowych typu trans (elaidynowego i wakcenowego) w mieku kobiecym.

INTRODUCTION: The structure of trans unsaturated fatty acids (TFAs) includes at least one double bond with a trans configuration. Numerous studies have shown that TFAs influence negative changes in blood lipid profile, may initiate or accelerate the course of many inflammatory diseases, and reduce n-6 and n-3 fatty acids' utilization. Their presence in tissues and systemic fluids is mainly the result of ingestion of TFA-containing foods. The objective of this study was to determine the concentration profiles of elaidic and vaccenic acids in human milk, and the existence of potential correlations between the levels of trans-unsaturated fatty acids and long chain polyunsaturated fatty acids. MATERIALS AND METHODS: The study group consisted of 53 lactating women, aged 18-39 years, 5-6 weeks after delivery. Elaidic and vaccenic acid levels were determined in human milk. Fatty acid methyl esters were injected onto the capillary column of an Agilent 6890M gas chromatography system integrated with an autosampler. Geometrical and positional isomers of fatty acids were identified by comparing their retention times with those of the fatty acid standards from Sigma-Aldrich. The content of individual fatty acids was determined from the reference curves and expressed in mg/mL. RESULTS: The level of elaidic acid was 0.2572 ± 0.1811 mg/mL, and the level of vaccenic acid was 0.2736 ± 0.1852 mg/mL. No negative correlations between the levels of the analyzed TFAs and PUFAs were observed in human milk. CONCLUSIONS: The levels of elaidic and vaccenic acid were similar and had no negative influence on the levels of arachidonic acid and docosahexaenoic acid.

[Overdosing of popular medicines available without a prescription, a new trend among teenagers]. / Naduzywanie popularnych leków dostepnych bez recepty nowym trendem wsród mlodziezy.

INTRODUCTION: For last several years popularity of substances which improve concentration or/and cause hallucinations has considerably increased. There is a conviction that most frequent addictions among teenagers are cigarettes, alcohol or drugs of abuse. A very dangerous phenomenon has been ignored, becoming more common among scholars and students - usage of medicines available without doctor's prescription (over the counter drugs - OTC). Popularity and fame of these specifics comes from side effects observed especially when overdosed. MATERIAL AND METHODS: In years 2007-2012 in Clinical Toxicology Laboratory of Medical Pomeranian University 4304 toxicological analyses was conducted with specified direction on drugs analysis (39% of all analyses made in that period). In some cases, especially of young people, a doctor ordering the analysis pointed as possible cause of intoxication "recreational usage" of OTC drugs. Those suspicions were confirmed by toxicological examinations. CONCLUSIONS: It seems that dominating character feature of people who abuse psychoactive substances is emotional immaturity. Maturation process can be impaired or completely inhibited by abusing psychoactive substances. This is processes are considered which enhance immaturity of person's personality - so called defense mechanisms.

Proteomic patterns analysis with multivariate calculations as a promising tool for prompt differentiation of early stage lung tissue with cancer and unchanged tissue material.

BACKGROUND: Lung cancer diagnosis in tissue material with commonly used histological techniques is sometimes inconvenient and in a number of cases leads to ambiguous conclusions. Frequently advanced immunostaining techniques have to be employed, yet they are both time consuming and limited. In this study a proteomic approach is presented which may help provide unambiguous pathologic diagnosis of tissue material. METHODS: Lung tissue material found to be pathologically changed was prepared to isolate proteome with fast and non selective procedure. Isolated peptides and proteins in ranging from 3.5 to 20 kDa were analysed directly using high resolution mass spectrometer (MALDI-TOF/TOF) with sinapic acid as a matrix. Recorded complex spectra of a single run were then analyzed with multivariate statistical analysis algorithms (principle component analysis, classification methods). In the applied protocol we focused on obtaining the spectra richest in protein signals constituting a pattern of change within the sample containing detailed information about its protein composition. Advanced statistical methods were to indicate differences between examined groups. RESULTS: Obtained results indicate changes in proteome profiles of changed tissues in comparison to physiologically unchanged material (control group) which were reflected in the result of principle component analysis (PCA). Points representing spectra of control group were located in different areas of multidimensional space and were less diffused in comparison to cancer tissues. Three different classification algorithms showed recognition capability of 100% regarding classification of examined material into an appropriate group. CONCLUSION: The application of the presented protocol and method enabled finding pathological changes in tissue material regardless of localization and size of abnormalities in the sample volume. Proteomic profile as a complex, rich in signals spectrum of proteins can be expressed as a single point in multidimensional space and than analysed using advanced statistical methods. This approach seems to provide more precise information about a pathology and may be considered in futer evaluation of biomarkers for clinical applications in different pathology. Multiparameter statistical methods may be helpful in elucidation of newly expressed sensitive biomarkers defined as many factors "in one point".

Death caused by addictive inhalation of nitrous oxide.

Intoxications with nitrous oxide have been, and still are, a rarity in forensic medicine. Apart from accidental overdose during hospital procedures, intoxication with this gas is the result of voluntary inhalation. We report the fatal case of a 32-year-old male who died during inhalation of nitrous oxide from whipped dairy cream cans and hint on the role of the internet in creating new behaviors among drug addicts. We rely on the autopsy report from the Department of Forensic Medicine, Pomeranian Medical University in Szczecin, on laboratory tests, and court files. Neither the autopsy nor the toxicologic and histopathologic tests disclosed the exact cause of death. However, circumstances in which the body was discovered were indicative that death resulted from cardiorespiratory failure. The present case is interesting with regard to its rarity, diagnostic difficulties and potential harm from nitrous oxide used by the food industry.

[Biochemical skin sweat-sebum profile in individual identification]. / Biochemiczny profil lojowo-potowy skóry w identyfikacji osobniczej.

INTRODUCTION: Sweat-sebum substance released by an individual, is left on the surface of different objects by direct contact (touch), which takes place in the well known mechanism of finger-print formation. Interesting from the crime detection point of view seems to be the determination of qualitative and quantitative changes in sweat-sebum biochemical profile left on objects, which may contribute to the determination of precise biochemical profiles in relation to individual identification. MATERIAL AND METHODS: Five different hand profiles of sweat-sebum were determined using gas chromatography with mass spectrometry obtaining referral material, then obtained profiles were compared to the profile left on a rough plastic surface by one individual belonging to the referral group. RESULTS: Obtained results indicate the possible application of the procedure for correlation of sweat-sebum skin profiles for identification purposes in crime detection, as indirect prove. Undoubtedly, although there is no possibility to show the uniqueness of particular profiles, like it is in a genetic examination, the short time needed to perform analysis (approximately 40 min) is very significant in the presented procedure, which may be a complementation of genetic methods in case were short time of perpetrator typing is a crucial step. CONCLUSIONS: Presented procedure may be helpful in preliminary individual identification in case were there is no possibility to collect finger-print profiles from the surface (inappropriate surface structure, smear traces).

[Lawsuit experiment as an important element in expert's work regarding carbon monoxide poisoning cases]. / Eksperyment procesowy jako istotny element w opiniowaniu zatrucia tlenkiem wiegla.

INTRODUCTION: Giving an opinion in the field of carbon monoxide intoxications is still problematic. MATERIAL AND METHODS: In the paper we present a case of the sudden death of a young female in the garage. In such a situation there were serious suspicions of a violent cause of the death. According to eye-witness relation an experiment to evaluate momentary carbon monoxide concentration in the garage was conducted. The toxicokinetic and biologic effects of carbon monoxide were taken under consideration. RESULTS: It was revealed that under the given parameters it was impossible there to reach a lethal carbon monoxide concentration.

[Diagnostics and interpretation of results in forensic toxicology of amphetamines]. / Diagnostyka i opiniowanie wyników w sadowo-toksykologicznej analizie amfetamin.

INTRODUCTION: Amphetamine and its analogues belong to the large group of psychoactive compounds. Due to the growing problem of amphetamine poisoning, author decided to review and improve the laboratory diagnostic procedures with regard to forensic toxicology analysis. MATERIAL AND METHODS: Biological material of 8 persons taken during autopsy for routine toxicological tests was used to examine characteristic of biological matrix. Following excludion the presence of xenobiotics during expertise process, material was divided and kept in sealed containers, then analysed after defined time period: 0, 30 and 90 days. Blood, urine, liver, brain and kidney were considered, 96 samples were examined in total. Biological material was analysed using gas chromatography equipped with flame ionization detector (GC-FID), liquid chromatography with diode array detector (HPLC-DAD), and magnetic resonance spectrometer (H NMR). CONCLUSIONS: 1. Workout analytical GC-FID method enable to verify positive initial result of amphetamine and metamphetamine analysis in urine. 2. Biological matrix is composed mainly of low molecular aliphatic organical compounds which cause significant impact in analytical and opinion problem in amphetamine analysis. Its influence determine in proportion as time of biological material self-decomposition and noted cross-reactivity concern both immunological and chromatographic methods (HPLC-DAD). 3. H NMR method may be used to confirm amphetamine presence in biological material for confirmation of positive results obtained using immunological and enzymatic methods used in forensic toxicology.