Quantification of MDMB-4en-PINACA and ADB-BUTINACA in human hair by gas chromatography–tandem mass spectrometry

Forensic Toxicology - To test synthetic cannabinoid (SCs) in parent forms from living human, the hairs seems to be one of the best samples, because of the non-invasiveness upon their collection....     

2-Methoxyqualone,a new recreational drug,discovered from a package seized by the police: a preliminary report

Forensic Toxicology -     

MALDI-TOF mass spectrometric analysis of α-amanitin, β-amanitin, and phalloidin in urine

A rapid and sensitive method was developed for analysis of α-amanitin, β-amanitin, and phalloidin by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this method, α-cyano-4-hydroxy cinnamic acid was used as the matrix to assist the ionization of toxins. The identification of α-amanitin, β-amanitin, and phalloidin was achieved through their sodium adducts [M+Na]⁺ at m/z = 941, 942, and 811, and quantification of the three toxins was also achieved using microcystin RR at m/z = 1038 as internal standard. For all toxins, the limit of detection was 5 ng/ml, and all calibration curves were linear in the range of 10–500 ng/ml using 0.4 ml of urine. The sensitivity for identification was increased about tenfold when the tandem MS (MS–MS) mode was used for detection. Because these quantifications could be achieved in the toxin concentration range of 4–200 ng, the present MALDI-TOF MS method can serve as the most sensitive method so far reported for the analysis of these mushroom toxins. To our knowledge, this study is the first trial to analyze amanitins and phalloidin by MALDI-TOF MS (-MS).     

Postmortem distribution of α-pyrrolidinovalerophenone and its metabolite in body fluids and solid tissues in a fatal poisoning case measured by LC–MS–MS with the standard addition method

We experienced an autopsy case, in which the cause of death was judged as α-pyrrolidinovalerophenone (α-PVP) poisoning. Other drugs or poisons that could have caused the death were not detected by our screening using gas chromatography–mass spectrometry. The deceased was a 41-year-old man. The postmortem interval was about 40 h. Cardiac blood, femoral vein blood, urine, stomach contents, and seven solid tissues were collected and frozen until analysis to investigate the distribution of α-PVP and its metabolite 1-phenyl-2-(pyrrolidin-1-yl)pentan-1-ol (OH-α-PVP) in the body of the cadaver. After sample pretreatment, they were subjected to solid-phase extraction with Oasis HLB cartridges and analysis by liquid chromatography–tandem mass spectrometry. Because the distribution study dealt with different matrices, we used the standard addition method to overcome the matrix effects. The concentration of α-PVP in urine was much higher than in other specimens; the concentrations of α-PVP in solid tissues except for the spleen were about twofold those in blood specimens. Among the solid tissues, the highest α-PVP concentration was found in the pancreas, followed by the kidney. The extremely high concentration of the drug in urine and the relatively high concentration in the kidney suggested that α-PVP is rapidly excreted into urine via the kidney. The distribution profile of OH-α-PVP was generally similar to that of α-PVP in body fluids and solid tissues. The concentration of OH-α-PVP in urine was also much higher than those in other specimens. Among the solid tissues, the OH-α-PVP concentration was highest in the liver, followed by the kidney. The high concentration of OH-α-PVP in the liver was expected, because the metabolism of α-PVP was probably most active in the liver. The high levels of OH-α-PVP in urine and kidney also suggested that the metabolite was also rapidly excreted into urine via the kidney. To test whether conjugated metabolites were present in urine and solid tissues, urine and homogenates of the liver and spleen were incubated with β-glucuronidase/sulfatase at 37 °C for 5 h. Concentrations of OH-α-PVP were measured before and after the incubation, but differences in the concentrations before and after the incubation were within 10 % for the urine, liver, and spleen samples. To date, data on the distribution of α-PVP and OH-α-PVP in body fluids and solid tissues in a fatal α-PVP poisoning case have not been reported; thus, the findings of our study will be useful for assessment of future cases of α-PVP poisoning.     

MALDI–TOF mass spectrometric determination of four amphetamines in blood

A rapid and sensitive detection method using matrix-assisted laser desorption ionization time-of-flight mass spectrometry was developed for the analysis of amphetamine (A), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA). In this method, α-cyano-4-hydroxy cinnamic acid was used as the matrix to assist the ionization of amphetamines. The MS spectra of these amphetamines showed protonated molecules [M + H]⁺ and fragment ions with comparable or higher intensities. The quantifications of A and MA were performed using A-d ₇ and MA-d ₅ as the internal standard, respectively, and those of MDA and MDMA were performed using MDMA-d ₅ as the internal standard. The limit of detection and the quantification range using 20 μl of blood were about 10 ng/ml and 30–500 ng/ml for A and MDA, respectively, and about 1 ng/ml and 3–50 ng/ml for MA and MDMA, respectively. In two cases of poisoning in which MA was abused, the levels of A in blood and urine were from 0.050 to 3.24 μg/ml and the levels of MA were from 0.231 to 25.1 μg/ml.     

MALDI-TOF mass spectrometric determination of four pyrrolidino cathinones in human blood

A rapid and sensitive detection method using matrix-assisted laser desorption ionization (MALDI)–time-of-flight (TOF)–mass spectrometry (MS) was developed for the analysis of four pyrrolidino cathinones: α-pyrrolidinopropiophenone (PPP), 4′-methyl-α-pyrrolidinopropiophenone (MPPP), α-pyrrolidinobutiophenone (PBP), and α-pyrrolidinovalerophenone (PVP). In this method, α-cyano-4-hydroxycinnamic acid was used as the matrix to assist ionization of the cathinones. Each MALDI–TOF–MS spectrum of the cathinones showed not only protonated molecular ion [M + H]⁺ but also several fragment ions having comparable intensities to that of [M + H]⁺. Hence, MPPP and PBP could be clearly discriminated by the mass spectra alone, although these compounds have almost the same mass numbers in their protonated molecular ions. The quantification of MPPP, PBP, or PVP was performed using PPP as internal standard, and that of PPP was performed using PBP as internal standard. The limit of detection was 1 ng/ml, and the quantification range was 2–200 ng/ml for the four cathinones using 20 μl of blood. In a fatal poisoning case in which PVP was abused, the PVP levels in whole blood samples obtained from the right heart, left heart, and femoral vein were 0.597, 0.635, and 0.580 μg/ml, respectively. We recommend the MALDI–TOF–MS method without any chromatography for both identification and quantification of the pyrrolidino cathinones in various matrices in forensic toxicological analysis, because of the simplicity, rapidness, and reliability of the method.     

Investigation on toxicological usefulness of synovial fluids,as an alternative matrix: postmortem distribution/redistribution of triazolam and its predominant metabolite α-hydroxytriazolam in human body fluids

Forensic Toxicology -     

Identification and quantitation of a new cathinone designer drug PV9 in an “aroma liquid” product,antemortem whole blood and urine specimens,and a postmortem whole blood specimen in a fatal poisoning case

A couple bought “aroma liquid” and “bath salt” type drugs at a dubious drug shop. Both of them orally took the liquid type drug; although the male subject showed no symptoms, the female subject suffered shivering, convulsions, and low levels of consciousness. The woman was taken to an emergency hospital to receive intensive medical treatment, but died about 20 h after admission. The aroma liquid solution, and the antemortem blood and urine collected during medical treatment at the hospital were brought to our laboratory by the police for analysis of the causative drug(s). In addition, a sample of postmortem femoral vein blood was collected from the cadaver. After some screening tests, we finally identified PV9 (α-POP) in all specimens by gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry (LC–MS–MS). The concentration of PV9 was 18.3 mg/ml in the aroma liquid solution, 45.7 ng/ml in the antemortem blood, 20.3 ng/ml in the antemortem urine, and 180 ng/ml in the postmortem femoral vein blood. The concentrations in antemortem blood and urine and in postmortem blood were greatly lowered by dilution during the intensive medical treatment, including intravenous drip infusion of a large volume of solution. The probable coexistence of a β-hydroxyl metabolite was also investigated by mass chromatography and analysis of fragment ions of the product ion spectrum obtained by LC–MS–MS. To our knowledge, this is the first reported identification and quantitation of PV9 in human specimens in a fatal PV9 poisoning case.