ABSTRACTThe QuEChERS extraction method was extensively modified and validated for effective screening of drugs and pesticides in diverse biological matrices, such as blood, urine, liver, and stomach contents. This method involved the extraction of 2 mL biological samples using acetonitrile as the solvent. To eliminate interferences, particularly lipids, a novel sorbent comprising a combination of C18 and specialized polymers called EMR-L (Enhanced Matrix Removal-Lipid) was employed. The extracted samples were then subjected to analysis using GCMS with a DB-5 MS column. The validation study encompassed various parameters including carryover, limit of detection (LOD), and interference investigation. LOD of analytes were ranged from 0.5 to 1.0 µg/mL. The validated method is known for its simplicity and effectiveness in conducting systemic toxicological analysis, covering a wide range of acidic, neutral, and basic analytes. Moreover, this method has the potential to detect additional drugs and pesticides not included in the validation study, provided they are compatible with gas chromatographic analysis. The modified method was successfully applied to real case samples, proving to be a valuable tool for systemic toxicological analysis. Its versatility allows for the screening of acidic, neutral, and basic drugs and pesticides in various biological matrices.KEYWORDS: QuEChERSEMRdrugspesticidespostmortemsystemic toxicologyunknown screen Disclosure statementNo potential conflict of interest was reported by the author(s).
Dextromethorphan is the most commonly used over-the-counter anti-tussive and expectorant medicine at therapeutic doses. Due to easy availability, euphoric high and hallucinogenic effects at larger doses, dextromethorphan popularity amongst the drug abusers is growing day by day. It is often mixed with alcohol, opiates, cannabinoids or other drugs of abuse for recreational purposes despite their lethal synergistic effects. More than 50 deaths were reported the first time in Pakistan after consuming cough syrups containing dextromethorphan, manufactured by two local pharmaceutical industries. All deceased had the history of drug abuse. We report the deaths of nineteen males, ages ranged from 17 to 45 years, in two major cities of Pakistan who purposefully ingested large doses of dextromethorphan for recreational purposes and died as a result of direct toxic effects of the drug. Toxicological analysis revealed high levels of dextromethorphan ranging from 7.3 to 41.7 mg/L in the peripheral blood, 4.2–92.6 mg/kg in the liver and 9.9–349.6 mg/L in the gastric content by high performance liquid chromatography. The dextromethorphan concentrations in all subjects significantly exceeded the therapeutic range and were consistent with concentrations reported in other cases of dextromethorphan abuse and toxicity. Besides dextromethorphan other drugs of abuse like cannabinoids, opiates, benzodiazepines, ethanol and chlorpheniramine were also detected. The cause of death was determined to be acute dextromethorphan intoxication with lethal synergistic effect of other co-ingested drugs of abuse. The deaths resulted in the prosecution of all individuals involved in manufacturing, distribution or sale of the cough syrup.
In January 2012, 664 cases of pyrimethamine toxicity and 151 deaths were reported among cardiac patients that had recently received free medicines from pharmacy of Punjab Institute of Cardiology, Lahore, Pakistan. These patients, ages ranged from 58 to 75 years, were prescribed simvastatin, clopidogrel, aspirin soluble, isosorbide mononitrate, and amlodipine. On examination of medications being given to them, it was found that a particular batch of isosorbide mononitrate tablets was contaminated with 50 mg pyrimethamine. Cardiac patients were taking isosorbide contaminated with pyrimethamine twice daily (100 mg pyrimethamine/day), whereas therapeutic dose of pyrimethamine for malaria is 25 mg/week. Postmortem urine, cardiac blood, and femoral blood specimens of three deceased males were submitted to author's laboratory for analysis. Postmortem toxicological analysis revealed that pyrimethamine concentration fell within the range of 1-10 μg/mL by liquid chromatography. Clinical, autopsy, histopathological, and toxicological findings strongly suggested toxicity due to pyrimethamine accumulation that resulted in deaths of these cardiac patients.
Astragalus psilocentros Fisch. is a medicinal plant belongs to family Papilionaceae and is used to cure various diseases such as cancer, diabetes, liver diseases, hypertension, cardic ischemia, heart failure, nephiritis etc.The aim of this study was to identify the antimicrobial activity for controlling pathogens and antioxidant efficacy.The methanolic extract of A psilocentros was dissolved in water and partitioned with hexane, chloroform, ethyl acetate and n-butanol successively.Phytochemical screening revealed the presence of alkaloids, terpenoids, tannins, phenolics, sugars, saponins and flavonoids.Antimicrobial activity was carried out against four bacterial (Bacillus subtilis, Pasturell multicoda, Escherichia coli, Staphylococcus aureus) and two fungal (Aspergillus niger and Alternia fusarium) strains.The chloroform, ethyl acetate and methanol extracts were effective against these tested microbes.The antioxidant potential of all these fractions and remaining aqueous fraction was evaluated by seven methods i.e.DPPH free radical scavenging activity, total antioxidant activity, FRAP assay, TPC assay, inhibition of lipid peroxidation, ABTS assay and super anion radical scavenging activity.All the fractions exhibited significant antioxidant potential.The results revealed that ethyl acetate showed the highest % inhibition by DPPH radical (183.92 ± 0.17), highest FRAP value of (56.86 ± 0.39), highest TPC value (53.69 ± 0.68) and highest inhibition by lipid peroxidation value (44.65 ± 0.44).The chloroform soluble fraction showed the highest TAA of (0.7005 ± 0.05).The GC-MS analysis of hexane soluble part of ethyl acetate fraction identified few potential markers responsible for the activity e.g., 1,3-bis-(trimethylsiloxy) benzene, diethyl benzoylmethylphosphonate, nonadecyl alcohol and 1,2-benzene dicarboxylic acid.
Cyanide is a deadly poison. Acute cyanide poisoning in humans is rare and is predominantly caused by smoke inhalation from fires and much more rarely by intentional ingestion of cyanide salts as in suicide or homicide attempts. The main objective of this report is to emphasize the need to consider cyanide poisoning, even if it is rare, in differential diagnosis while evaluating cases of sudden death and the significance of gastric content analysis in acute poisoning cases. The authors reported four cases of lethal cyanide poisoning. Autopsy specimens were submitted to the author’s laboratory for analysis. A presumptive test for cyanide using Vitamin B12 indicated the presence of cyanide. Confirmation and quantification of cyanide was performed using headspace gas chromatograph coupled to flame ionization detector technique. The toxicological analysis revealed lethal hydrogen cyanide concentrations in all postmortem specimens ranging from 24 to 2600 mg/L in gastric contents, 70 to 282 mg/kg in liver specimens, 11 to 12 mg/kg in a mixture of viscera (liver, spleen, kidney) and 15 mg/L in blood. The cause of deaths in the reported cases was acute respiratory failure and cardiac arrest following cyanide intoxication.
Abstract A rapid colorimetric method for detection of p‐phenylenediamine ( PPD ) in various biological samples is developed. The o‐cresol test for acetaminophen detection has been modified to detect PPD in blood, urine, gastric contents, and liver. After precipitating protein with trichloroacetic acid solution (2 mL, 10% w/v), biological specimens were required to convert PPD metabolites to PPD by acid hydrolysis. Finally, o‐cresol solution (1 mL, 1% w/v), hydrogen peroxide (200 μL, 3%v/v), and concentrated ammonium hydroxide (0.5 mL) were added in the biological samples. The presence of PPD was indicated by formation of violet color which was turned to bluish green color within 10–15 min. The limit of detection was found to be 2 mg/L in blood, urine, and gastric contents and 2 mg/Kg in liver. This method is also free from any potential interference by p‐aminophenol, acetaminophen, and other amine drugs under test conditions. This method was successfully employed to thirteen fatal cases of PPD poisoning.
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