lachrymal fluid as an alternative biological matrix for toxicological purpose

Abstracts / Toxicology Letters 229S (2014) S40–S252 S99

diazepine level was positive according to fluorescence polarizationimmunoassay method and toxicological screening was negative forother drugs. Ten hours after ingestion she developed impressiveopisthotonus and deep coma that required mechanical ventilation.Brain tomography was in normal ranges. CSF features included clearaspect, normotensive, without biochemical abnormalities. After48 h of nonresponsive evolution a new brain CT scan identifiedmassive cerebral edema. Brain MRI showed hyperintensities onT2 sequences in basal ganglia bilaterally and diffusion restrictionin these area suggesting progressive multifocal leukoencephalopa-thy. Considering these aspect we performed PCR for the detectionof JC virus which was negative. A prionic disease was highly sus-pected and 14-3-3 protein immunoassay on CSF was positive. Thepatient died after 2 months in Critical Care Unit due to multipledysfunctions.

Conclusions: Our case underlines the importance of keeping anopen mind when dealing with unexpected and unusual evolutionof acute intoxication.

http://dx.doi.org/10.1016/j.toxlet.2014.06.364

P-2.20Suicide attempt by pharmaceutical ingestion ina tertiary unit

Radu Tincu ∗, Radu Macovei, Zoie Ghiorghiu

Bucharest Clinical Emergency Hospital, Bucharest, Romania

Background and aims: Poisoning is a common way of suicideattempt, causing important mortality and morbidity. Our objectiveis to describe the characteristics of patients’ presenting for suicideattempt through substance abuse over 1 year period.

Material and methods: We performed a prospective cross sec-tion study in our Intensive Care – Toxicology Unit between January2013 and January 2014. All suicide attempts through drug abusewere registered. Each admitted patient underwent a completequestionnaire that included: age, sex, living area, living standards,profession, motivation, type of the substance, number of attempts,comorbidities, psychiatric evaluation.

Results: A total of 416 cases were analyzed: 263 women, 153men, mean age 43.9 years. The pattern of exposure resulted: antibi-otics (1.9%), no steroidal anti-inflammatories (7.84%), vitamins(0.98%), sedatives (13.7%), hormone pills (0.98%), antidepresives(6.86%), anti H2 receptors (2.94%), amphetamines (0.98%), anti-hypertensive (2.94%), antiepileptic (5.88%), beta blockers (1.96%),plants (0.98%), antipsychotics (7.84%), acetaminophen (7.84%), rati-cids (1.96%), etnobotanics (1.96%), benzodiazepines (32.35%). Theleading cause of suicidal act was economic problems and unem-ployment (p < 0.005). Patients belong to different social and culturallevels, but we report suicidal attempts in prisoners (5 cases) andnouns (3 cases). Death was registered in 3 cases and it was due torespectively hepatic acute failure and multiple organ dysfunctions.

Conclusions: Knowing the cause of suicidal attempt may improveprevention. This study shows that financial problems during eco-nomic crisis and domestic conflicts are the most important triggersfor suicidal act. In the meantime, particular social layers were rep-resented amongst our patients such as prisoners and nouns.

http://dx.doi.org/10.1016/j.toxlet.2014.06.365

P-2.21Lachrymal fluid as an alternative biologicalmatrix for toxicological purpose

Renata Wietecha-Posłuszny ∗, Agnieszka Moos, AgnieszkaBochenska, Paweł Koscielniak

Laboratory for Forensic Chemistry, Department of AnalyticalChemistry, Jagiellonian University, Krakow, Poland

The aim of this study was to apply human lachrymal fluidas an unusual biological matrix to isolation of five benzodi-azepines (BZD): tetrazepam, clonazepam, estazolam, diazepam andalprazolam by semi-automated microextraction by packed sor-bent (eVol®MEPSTM). All samples were collected from healthyvolunteers via free spillage on a cheek to disposable test tubes.Analyses were performed using a ultra-high performance liq-uid chromatography technique coupled with mass spectrometrydetector (UHPLC–MS-TOF). A lachrymal fluid samples were cen-trifuged just prior to the MEPS extraction. Moreover, it was shortand simple to perform with limited sample consumption (only30 �L).

The MEPS/UHPLC–MS-TOF method was validated at threeconcentration levels: 4.00, 8.00 and 12.00 ng/mL of all BZD ana-lytes in the range 2–15 ng/mL. The following parameters weredetermined: limit of detection, limit of quantification, linearity,precision, accuracy and recovery. Obtained LOQ values, rangingfrom 0.03 to 0.09 ng/mL, made the method to be a useful tool inforensic laboratories, which was demonstrated on the basis ofanalysis of real samples.

Acknowledgments: A. Moos gratefully acknowledges the finan-cial support from the project Interdisciplinary PhD Studies“Molecular Sciences for Medicine”.

The research was carried out with equipment purchased thanksto the financial support of POIG.02.01.00-12-023/08).

http://dx.doi.org/10.1016/j.toxlet.2014.06.366

P-2.22Metabolomics and miRNA biomarkers ofparacetamol overdose in rodents and children

Richard Beger 1,∗, Xi Yang 1, Lisa Pence 1, Jinchun Sun 1, LauraSchnackenberg 1, William Salminen 2, Qiang Shi 1, JamesGreenhaw 1, Prit Gill 3, Sudeepa Bhattacharyya 3, Pippa Simpson 4,Kan Ye 4, Donna Mendrick 1, Laura James 3

1 NCTR, US FDA, Jefferson, AR, USA, 2 PAREXEL International, Sarasota,FL, USA, 3 Arkansas Children’s Hospital Research Institute, Little Rock,AR, USA, 4 Medical College of Wisconsin, Milwaukee, WI, USA

The commonly used drug, paracetamol, also known asacetaminophen (APAP), is a major cause of liver injury in adultsand children. APAP-induced toxicity causes oxidative stress, ROSproduction, glutathione depletion, mitochondria dysfunction, dis-ruption of energy metabolism, and altered immune response.Biofluid and tissue samples from preclinical APAP toxicity studieswere analyzed by genomics, proteomics and metabolomics tech-nologies to discover omics biomarkers of liver injury. Some ofthe metabolomics and miRNA biomarkers that were found in thepreclinical studies have been evaluated in blood and urine sam-ples from children that were overdosed with APAP. Metabolomicsevaluations of samples from a preclinical study of APAP toxicityidentified changes related to fatty acid beta-oxidation metabolismand bile acids transportation. In a subset of clinical samples,