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This study aimed to compare whole blood and serum concentrations of quetiapine in acute poisoning cases. Authentic whole blood and respective serum samples were routinely collected from patients diagnosed with blood poisoning at our University Hospital. Accordingly, whole blood and serum paired samples from nine patients (one male and eight female patients) were analyzed for quetiapine using liquid chromatography-mass spectrometry (LC-MS). Quetiapine concentrations in whole blood and serum samples ranged widely from 5.4 to 2780 ng/mL and 9.9 to 2500 ng/mL, respectively. The whole blood/serum concentration ratio was 0.5-1.1 and increased together with an increase in whole blood and serum quetiapine concentrations. The ratio was reversed at around 2500 ng/mL to 1. Our findings suggest that whole blood concentrations are more useful than serum concentrations in diagnosing quetiapine poisonings.

Azacyclonol metabolized from fexofenadine and pipradrol are structural isomers and are important biomarkers to determine fexofenadine or pipradrol intake. Detecting these two analytes by liquid chromatography–mass spectrometry (LC–MS) allows for their differentiation from structurally similar compounds. This study aimed to confirm the separation of azacyclonol and pipradrol through different chromatographic conditions in LC–MS. Urine samples were prepared by HLB and were separated in a high-performance reversed phase column. MS detection was achieved using a triple quadrupole mass spectrometer in positive ionization modes. Azacyclonol and pipradrol were differentiated by optimizing the gradient profile using LC–MS. We used a common Poroshell C18 column with acetonitrile and 0.1% acetic acid solution. Azacyclonol and fexofenadine were detected in the urine of patients who took fexofenadine. Fexofenadine is an important biomarker for the determination of azacyclonol and piradrol. LC–MS has been the dominant analytical technology for the quantitative bioanalysis of drugs and metabolites. Although some analytes do not require sufficient separation by LC according to selectivity by LC–MS, sufficient separation by LC is required for the separation of such position isomers.

Purpose Lisdexamfetamine (LDX), which is used for the treatment of attention-deficit/hyperactivity disorder and narcolepsy, is composed of l-lysine attached to dextroamphetamine (d-amphetamine). In this article, we report a forensic autopsy case in which prescription drugs were unknown at autopsy. While amphetamine was detected, methamphetamine could not be detected by liquid chromatography–tandem mass spectrometry (LC–MS/MS) in any of samples collected. Thus, we aimed to quantify LDX concentrations in autopsy samples and to prove that the amphetamine detected in this case was due to metabolized LDX. Methods Femoral vein blood, cardiac whole blood, urine, and gastric content samples were taken at autopsy for toxicological analysis. Qualitative and quantitative analyses were performed using LC–MS/MS. In addition, optical isomer separation for the amphetamine detected was conducted. The stability of LDX in whole blood and urine was also examined at three different temperatures. Results The concentrations of LDX were d-amphetamine by optical isomer separation. The d-amphetamine detected was considered to be derived from LDX. Stability experiments revealed that LDX in whole blood decreased at ambient temperature. Conclusions The results in the present case report may be useful in interpreting whether or not the amphetamine detected in a cadaver is a metabolite of LDX.

We developed and validated an assay for determination of glyphosate (GLYP) and glufosinate (GLUF) in human serum. Serum samples were extracted by using a MonoSpin® TiO column and analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). MonoSpin® TiO tends to specifically bind to phosphate groups. The assay was linear over a concentration range of 1–250 μg/mL. The recoveries for the 2 compounds were 1.6%–2.3%. The intra- and inter-day variations were

A novel method coupling spin column extraction with high-performance liquid chromatography–mass spectrometry was developed for simultaneous extraction of β-blockers and calcium channel blockers fro...

Cyanide is a powerful and rapidly acting poison. In Japan, cyanide poisoning is rare, and regular cyanide testing can be costly and time consuming. In contrast, alcohol analysis is routinely performed in most forensic laboratories. In this study, we attempted to develop a method for the simultaneous quantification of cyanide and alcohols in blood using headspace gas chromatography (HS-GC). As nitrogen-phosphorus detection (NPD) is more sensitive to hydrogen cyanide than mass spectrometry (MS), a Deans switch was used to switch the detectors during a single run. The separation provided by three analytical columns, PoraBOND Q, CP-Sil 5 CB, and HP-INNOWax, was investigated, and PoraBOND Q was selected. The use of HS-GC-MS/NPD with a Deans switch enabled the simple and simultaneous quantification of cyanide, ethanol, and 1-propanol. Eighteen other volatile compounds were detected in the SIM/scan mode of the MS.

A simple and cost-effective method for analyzing valproic acid (VPA) in biological samples was developed. VPA was extracted in methyl tertiary-butyl ether (MTBE) and derivatized using trimethylsilyldiazomethane. The MTBE extract was analyzed by gas chromatography-mass spectrometry (GC-MS). The extraction recovery in human whole blood and urine was over 90 %, with good linearity in the range of 1.0 to 250 µg/mL of VPA. The RSD for 2.0, 20, and 200 μg/mL VPA in whole blood ranged from 0.9 to 4.7 % for intra-day and 1.5 to 5.9 % for inter-day. The RSD for 2.0, 20, and 200 μg/mL VPA in urine ranged from 1.9 to 2.6 % for intra-day and 1.2 to 2.9 % for inter-day. As a preliminary cross-validation study, a cross-check was conducted using blinded concentration samples. The results demonstrated that the assay data of the two laboratories were comparable.

Although organophosphorus agents are used worldwide as pesticides, there have been many reports of pesticide poisoning. Nerve agents are organophosphorus agents that interfere with neurotransmission and have been used as chemical weapons in wars. These agents mainly irreversibly inhibit the action of acetylcholinesterase, an enzyme that breaks down acetylcholine, a neurotransmitter, and are believed to cause acute symptoms of poisoning. However, in recent years, the presence of subacute, delayed toxicity independent of acetylcholinesterase inhibition has been reported for some organophosphorus agents. We analyzed the subacute and delayed toxicity of bis(isopropylmethyl)phosphonate (BIMP), which has the same phosphonate group as sarin. BIMP rounded out the morphology of the cells and reduced the proportion of cells in the G1 phase of the cell cycle over time. No DNA damage was observed, suggesting that BIMP may affect cell division.

Objectives The aim of this study was to develop and validate a simple and reliable gas chromatography‐mass spectrometry (GC‐MS) method to simultaneously determine urinary 1‐naphthol (1‐NAP) and 2‐naphthol (2‐NAP) for biological monitoring of occupational exposure to naphthalene. Methods NAPs were derivatized in situ with acetic anhydride after enzymatic hydrolysis, extracted with n‐hexane, and analyzed using GC‐MS. Validation of the proposed method was conducted in accordance with US Food and Drug Administration guidance. A final validation was performed by analyzing a ClinChek®‐Control for phenolic compounds. Results The linearity of calibration curves was indicated by a high correlation coefficient (>0.999) in the concentration range 1‐100 μg/L for each NAP. The limits of detection and quantification for each NAP were 0.30 and 1.00 μg/L, respectively. The recovery was 90.8%‐98.1%. The intraday and interday accuracies, expressed as the deviation from the nominal value, were 92.2%‐99.9% and 93.4%‐99.9%, respectively. The intraday and interday precision, expressed as the relative standard deviation, was 0.3%‐3.9% and 0.4%‐4.1%, respectively. The ClinChek® values obtained using our method were sufficiently accurate. Conclusions The proposed method is simple, reliable, and appropriate for routine analyses, and is useful for biological monitoring of naphthalene exposure in occupational health practice.

We have previously reported that ischemic animal models treated with a respiratory inhibitor, rotenon, show an increased voluntary alcohol intake. Although it is clear that ischemic brain, as a result of reduced-blood flow, shows pathological events and/or neuro-degenerations apparently, little is known of causal relationship between the mechanism of neural dysfunction and voluntary alcohol consumption. Authors have investigated effects of permanent two-vessel occlusion (p2VO) on rat voluntary alcohol drinking behavior. In first experiment the p2VO-treated rats showed an increase of voluntary alcohol drinking behavior, as compared with sham controls. Using brain microdialysis technique, increases of only nucleus accumbens (ACC) dopamine (DA) releases were suppressed in the p2VO-treated rats significantly, following the high K+ (40 mM) perfusion through the microdialysis probe membrane. Alcohol (200 mM) perfusion-induced DA and serotonin (5-HT) releases in the ACC of the p2VO-treated rats were suppressed significantly in the second experiment, as compared with the sham-treated rats. In third experiment p2VO-treated rats showed significant decreases of the contents of DA, not 5-HT, in the ACC, caudate-putamen (C/P), ventral tegmental area-substantia nigra (VT/SN) and lateral hypothalamus (LH). Dopaminergic neurons in the ACC showed more functional vulnerability against the p2VO treatments, as compared with the serotonergic neurons. An increase of alcohol intake in the p2VO-treated rats means the compensation for the neural degeneration of the dopaminergic system in the ACC consisted brain rewarding system. It was likely suggested that neural disturbance of higher functions involved with incomplete global brain ischemia leads the risk of an abnormal alcohol drinking in human.

This study attempted to determine the phenothiazine antipsychotics concentration in serum and whole blood samples using various diatomaceous earth-based solid-phase columns and elution solvents and subsequently evaluate their efficiency. Phenothiazine antipsychotics concentrations of 5 - 2000 ng/mL were extracted from serum and whole blood using each column. All compounds were analyzed using liquid chromatography-tandem mass spectrometry. Phenothiazine antipsychotics extraction in serum and whole blood using diatomaceous earth-based solid-phase columns seemed to have an affinity with the elution solvent.

This article describes analytical methods for determining alcohol levels in body fluids of living and dead persons. Widmark microdiffusion and Conway cell microdiffusion methods utilize the reduction of dichromate ion to chromium ion by ethanol. Several commercial kits utilize the enzyme ADH activity to reduce NAD + to NADH; EMIT, an automated instrument, utilizes the enzyme glucose-6-phosphate dehydrogenase activity, instead of ADH. Gas–liquid chromatography is the most commonly used method for forensic and clinical purposes. For breath alcohol testing, many on-site instruments utilizing infrared spectroscopy technology and/or electrochemical oxidation/fuel cell are available.

OBJECTIVES The aim of the present study was to comparatively evaluate the usefulness of urinary cyclohexanediols (CHdiols-U) and cyclohexanol (CHol-U) as biomarkers of occupational exposure to cyclohexane (CH). METHODS Sixteen subjects (14 men and 2 women) were exposed to CH during proof-printing work. Personal exposure monitoring was conducted during the whole shift on the last working day of the week. The time-weighted average level of exposure to CH (CH-A) was measured using a diffusive sampler. Two urine samples were collected from each worker at different times during the same week: a baseline urine sample (before the first shift of the working week, after a 5-day holiday with no CH exposure) and an end-of-shift urine sample (after the last shift of the same working week, the same day personal exposure monitoring was conducted). CH-A, CHdiols-U and CHol-U were determined using a gas chromatograph-flame ionization detector. RESULTS The CH-A concentrations ranged from 4.5 to 60.3 ppm, with a geometric mean (GM) of 18.1 ppm. The GMs and ranges (in parenthesis) of the creatinine (cr)-corrected end-of-shift 1,2-CHdiol-U, 1,4-CHdiol-U and CHol-U concentrations were 12.1 (4.1-36.6), 7.5 (2.4-20.1) and 0.4 (0.2-1.0) mg/g cr, respectively. Both CHdiols-U at the end of the shift were significantly correlated with CH-A (correlation coefficients for 1,2-CHdiol-U and 1,4-CHdiol-U of 0.852 and 0.847, respectively). No correlation was observed between CH-A and CHol-U. CONCLUSIONS CHdiols-U at the end of the last shift of the working week are suitable biomarkers of occupational exposure to CH, but CHol-U is not suitable.

Aconitum and Colchicum are known to be highly toxic plants based on their respective toxins, namely aconite and colchicine. Aconite and colchicine poisoning generally occurs because of misidentification and contamination, and is also associated with homicide and suicide cases. The detection of aconitines and colchicine in body fluids and leftovers plays an important role in clinical and forensic toxicology investigation of suspected food poisoning by natural poisons. Therefore, we have developed and validated a rapid determination of aconitine, hypaconitine, jesaconitine, mesaconitine, and colchicine in serum, urine, and plants using liquid chromatography–tandem mass spectrometry. All targeted compounds, together with internal standards, were extracted from serum and urine using a C18 monolithic spin column. Recovery of all compounds from serum, spiked at concentrations of 0.1, 0.8, 6, and 18 ng mL−1, ranged from 96.8 to 101.4 %. Recovery of all compounds from urine, spiked at concentrations of 0.1, 8, 80 and 190 ng mL−1, ranged from 96.6 to 101.9 %. The intra- and inter-assay accuracy and precision (expressed as relative standard deviation, %RSD) evaluated for all compounds were within 96.7–103.2 % and 1.7–11.5 %, respectively. Detection limits and quantitation limits of aconitines and colchicine for serum and urine were 0.05 and 0.1 ng mL−1, respectively. This method had a chromatographic run time of 7.0 min. Calibration curves for serum and urine ranged from 0.1 to 20 ng mL−1 for serum and 0.1–200 ng mL−1 for urine, respectively. The validated method was successfully applied to clinical aconitine and colchicine poisoning cases.

A number of N-alkyl indole or indazole-3-carbonyl analogs, with modified chemical structures, are distributed throughout the world as synthetic cannabinoids. Like synthetic cannabinoids, cathinone analogs are also abused and cause serious problems worldwide. Acute deaths caused by overdoses of these drugs have been reported. Various analytical methods that can cope with the rapid changes in chemical structures are required for routine analysis and screening of these drugs in seized and biological materials for forensic and clinical purposes. Although many chromatographic methods to analyze each drug have been published, there are only a few articles summarizing these analytical methods. This review presents the various colorimetric detections, immunochemical assays, gas chromatographic–mass spectrometric methods, and liquid chromatographic–mass spectrometric methods proposed for the analysis of synthetic cannabinoids and cathinones.

Monolithic silica in MonoSpin for solid-phase extraction of drugs from whole blood samples was developed to facilitate high-throughput analysis. Monolithic silica of various pore sizes and octadecyl contents were synthesized, and their effects on recovery rates were evaluated. The silica monolith M18-200 (20μm through-pore size, 10.4nm mesopore size, and 17.3% carbon content) achieved the best recovery of the target analytes in whole blood samples. The extraction proceeded with centrifugal force at 1000rpm for 2min, and the eluate was directly injected into the liquid chromatography-mass spectrometry system without any tedious steps such as evaporation of extraction solvents. Under the optimized condition, low detection limits of 0.5-2.0ngmL-1 and calibration ranges up to 1000ngmL-1 were obtained. The recoveries of the target drugs in the whole blood were 76-108% with relative standard deviation of less than 14.3%. These results indicate that the developed method based on monolithic silica is convenient, highly efficient, and applicable for detecting drugs in whole blood samples.

A simple and precise drug screening method was developed for the detection of abused drugs in whole blood by using the Oratect® III device that is usually employed for the detection of drugs in saliva. Whole blood was acidified with phosphoric acid, following which the hemolyzed solution was filtered through the ultrafiltration column Vivaspin 2 Hydrosart®. The filtrate was then tested for the presence of drugs using Oratect III. The detection limit of the device for methamphetamine, amphetamine, morphine, codeine, dihydrocodeine, diazepam, alprazolam, estazolam, and prazepam in whole blood was 125, 125, 50, 50, 50, 25, 60, 15, and 75ng/mL, respectively. The concentration range detected was between therapeutic and toxic drug levels; therefore, the proposed method can be applied for detecting the presence of abused drugs in blood. Our method is a novel, optimized technique for use in forensic laboratories to screen whole blood for drugs of abuse.

Clinical diagnosis of tetrodotoxin (TTX) poisoning can be difficult because of the lack of characteristic morphological findings and a screening test, such as an immunoassay. Here, we present a fully validated method for the analysis of TTX in serum and urine. In this method, serum and urine samples were extracted using MonoSpin CBA or amide columns, followed by LC–MS/MS analysis. The TTX was eluted from the column by 0.1 mL of 10 % acetic acid solution, and was directly injected into LC–MS/MS. An Agilent 1200 HPLC system equipped with a HILIC separation column (Zorbax HILIC Plus 2.1 × 100 mm, 3.5 μm) was used for isocratic elution, with a mobile phase of 10 mM ammonium formate with formic acid (95:5, v/v), along with 5 mM trifluoroacetic acid and 2 % acetonitrile. TTX was detected with an Agilent 6410 mass spectrometer utilizing positive electrospray ionization and multiple reaction monitoring. Limits of quantification for serum and urine were established to be 1 and 0.5 ng mL−1, respectively. Limits of detection for serum and urine were 0.5 and 0.25 ng mL−1, respectively. Intra-day and inter-day precision varied from 1.5 to 8.5 %. The recovery was >86.5 % for both matrices. In this method, the sample preparation process prior to injection into the LC–MS/MS takes approximately 10–15 min, which reduces the extraction time to one-tenth of that of previous methods. The application of this method was further verified by analysis of biological materials from a patient suffering from puffer fish poisoning.

For detection of a history of drug abused, we developed a simple method for extracting pyrrolidinophenone-type designer drugs in human hair by using a MonoSpin(®) C18 column. Target drugs were extracted from a single alkaline-digested hair segment (length, 10mm; weight, ca 0.1mg). The analytes extracted were then analyzed by high-performance liquid chromatography-mass spectrometry without evaporation of the eluent after MonoSpin extraction. Linearity from 0.5 to 500ng/mg was observed for all the tested drugs using an internal standard method (correlation coefficients >0.998) and the limit of detection was 0.2ng/mg. The recoveries were between 0.7 and 11.1%. The coefficients for intraday and interday variations at 4, 40, 200, and 400ng/mg in hair were between 0.7 and 11.1%. This method was successfully applied to the identification of these designer drugs in segmented human hair from drug abusers and indicated their history of drug abuse. The results were consistent with the patients' statements, indicating that this rapid method can be used to detect a history of drug abuse.

The organophosphorus compound sarin irreversibly inhibits acetylcholinesterase. We examined the acute cardiovascular effects of a sarin-like organophosphorus agent, bis(isopropyl methyl)phosphonate (BIMP), in anaesthetized, artificially ventilated rats. Intravenous administration of BIMP (0.8mg/kg; the LD50 value) induced a long-lasting increase in blood pressure and tended to increase heart rate. In rats pretreated with the non-selective muscarinic-receptor antagonist atropine, BIMP significantly increased both heart rate and blood pressure. In atropine-treated rats, hexamethonium (antagonist of ganglionic nicotinic receptors) greatly attenuated the BIMP-induced increase in blood pressure without changing the BIMP-induced increase in heart rate. In rats treated with atropine plus hexamethonium, intravenous phentolamine (non-selective α-adrenergic receptor antagonist) plus propranolol (non-selective β-adrenergic receptor antagonist) completely blocked the BIMP-induced increases in blood pressure and heart rate. In atropine-treated rats, the reversible acetylcholinesterase inhibitor neostigmine (1mg/kg) induced a transient increase in blood pressure, but had no effect on heart rate. These results suggest that in anaesthetized rats, BIMP induces powerful stimulation of sympathetic as well as parasympathetic nerves and thereby modulates heart rate and blood pressure. They may also indicate that an action independent of acetylcholinesterase inhibition contributes to the acute cardiovascular responses induced by BIMP.

We report the time-course profile of urinary excretion of α-pyrrolidinovalerophenone (α-PVP) and α-pyrrolidinobutiophenone (α-PBP) after intravenous injection in a human. The subject was a man aged in his forties, who intravenously injected unknown amounts of so-called unregulated drugs with the intention to commit suicide. The urinary concentrations of the drugs were analyzed by MonoSpin® extraction and gas chromatography–mass spectrometry. α-PVP, α-PBP, and their 2′-oxo metabolites of these compounds were identified in the urine. The highest concentration of α-PVP and α-PBP in urine was 1.2 and 1.6 μg/ml, respectively. The excretion half-lives of α-PVP and α-PBP were calculated to be 22 and 11 h, respectively. To our knowledge, this is the first report on the urinary excretion profiles of α-PVP and α-PBP in a human.

We report a fatal case of 3,4-methylenedioxypyrovalerone (MDPV) poisoning. A 35-year-old woman was taken to a nearby hospital after she was found unconscious. MDPV (1,200 ng/ml) and α-pyrrolidinobutiophenone (α-PBP; 200 ng/ml) were detected in her cardiac blood. Hair segments from the victim were also analyzed to confirm chronic drug use. MDPV and α-pyrrolidinovalerophenone (α-PVP) were found in many of the segments tested with the highest MDPV concentration at 22 ng/10-mm hair. We concluded that the woman died of acute MDPV poisoning, and that she had abused MDPV and α-PVP for a long time. To our knowledge, this is the first report dealing with MDPV in human hair segments, and with α-PBP in human specimens.

Organophosphates and carbamates are pesticides whose acute toxicities are caused by inhibition of acetylcholinesterase. A liquid chromatography–mass spectrometry/mass spectrometry method was developed and validated for the quantification of 16 organophosphate and carbamate insecticides in human serum. Nonpolar and polar pesticides were simultaneously extracted from serum samples using a simple and fast monolithic spin column procedure using mixed-mode C-C18 cartridges. Recovery was achieved in the range 11.9–99.2 %. Chromatography was carried out on an InertSustain® C18 HP 3 μm analytical column with gradient elution. Mass spectrometric analysis was performed using an Agilent 6410 Triple Quad Tandem mass spectrometer coupled with an electrospray ionization source in the positive ion mode. The assay was validated over a large concentration range and the limits of detection for all compounds ranged from 0.1 to 50 ng mL−1. The precision for both intra- and inter-day determination of all analytes was found to be acceptable (

A 59-year-old man was found dead in his house, where three sachets containing herbal blends were found on a table. The sachet contents were analyzed by gas chromatography–mass spectrometry and found to contain [1-(5-fluoropentyl)-1H-indol-3-yl](4-methyl-1-naphthalenyl)methanone (MAM-2201). The deceased was subjected to forensic autopsy. There were neither external injuries nor endogenous diseases judged by macroscopic and microscopic observations. Liquid chromatography–electrospray ionization–tandem mass spectrometry was used to quantitate the concentrations of MAM-2201 in postmortem samples using deuterated MAM-2201 as internal standard. The MAM-2201 concentrations were: 12.4 ng/ml in whole blood; 18.1 ng/g in the liver; 11.2 ng/g in the kidney; 4.3 ng/g in the brain; and 1,535 ng/g in the adipose tissue. We concluded that the man’s death was caused by acute intoxication with MAM-2201. In addition, we propose that the adipose tissue is the specimen of choice to detect MAM-2201 in the unchanged form. To our knowledge, this is the first report of a fatal MAM-2201 poisoning case. In addition, this report is also the first to describe the distribution of the drug in postmortem human tissues and blood.

The extraction and/or purification of drugs and medicines from biological matrices are important objectives in investigating their toxicological and pharmaceutical properties. Many widely used methods such as liquid–liquid extraction or SPE, used for extracting, purifying and enriching drugs and medicines found in biological materials, involve laborious, intensive and expensive preparatory procedures, and they require organic solvents that are toxic to both humans and the environment. Recent trends are focused on miniaturization, high-throughput and automation techniques. All the advantages and disadvantages of these techniques and devices in biological analysis are presented, and their applications in the extraction and/or purification of drugs and medicines from biological matrices are discussed in this review.

We provided toxicological analytical support for a fatal case of abuse of α-pyrrolidinovaleorophenone (α-PVP). Solid-phase microextraction (SPME) and capillary gas chromatography coupled to mass spectrometry (GC–MS) was employed to quantify the drug in whole blood. The whole blood concentration of the drug in the heart was 486 ng/ml. This is the first report of α-PVP intoxication as ascertained by mass spectrometric identification of α-PVP in whole blood.

A 42-year-old man presented with coma from methanol intoxication (178 mg/dL; its metabolite, formic acid, 860 μg/mL). Continuous hemodiafiltration improved his disturbance of consciousness, but he subsequently developed cognitive dysfunction and parkinsonism, which were finally alleviated by rehabilitation and amantadine therapy. Brain MRI revealed acute basal ganglionic and hippocampal lesions and chronic developmental white matter lesions (figures 1 and 2).

Organophosphorus (OP) compounds such as sarin are toxic agents that irreversibly inhibit the enzyme acetylcholinesterase. A recent study showed that OP compounds also have multiple toxicity mechanisms, and another suggested that endoplasmic reticulum (ER) dysfunction contributes to OP toxicity. However, the signaling pathway and mechanisms involved are poorly understood. We examined whether the sarin-like OP agent bis(isopropyl methyl)phosphonate (BIMP), which exhibits toxicity similar to that of sarin, induced ER stress in human astrocytoma CCF-STTG1 cells. Our results demonstrate that BIMP exposure reduced cell viability. Moreover, it induced changes in mitochondrial membrane potential and increased cleavage of caspase 3. Treatment with BIMP increased the mRNA levels of the ER stress marker genes binding immunoglobulin protein (BiP) and the transcription factor C/EBP homologous protein (CHOP). Furthermore, BIMP increased the protein expressions and phosphorylation of BiP, CHOP, and protein kinase RNA-like ER kinase and the phosphorylation of eukaryotic translation initiation factor 2. Compared to BIMP treatment alone, pretreatment with the CHOP siRNA, siCHOP, decreased BIMP-dependent CHOP expression and improved CCF-STTG1 cell viability. Our findings suggest that BIMP induced mitochondrial dysfunction and apoptotic cell death event mediated by ER stress in CCF-STTG1 cells and that treatment targeted at managing ER stress has the potential to attenuate the toxicity of OP nerve agents.

Organophosphorus compounds, such as sarin, are highly toxic nerve agents that inhibit acetylcholinesterase (AChE), but not cholinesterase, via multiple mechanisms. Recent studies have shown that organophosphorus compounds increase cyclooxygenase-2 (COX-2) expression and induce neurotoxicity. In this study, we examined the toxicity of the sarin-like organophosphorus agent bis(isopropyl methyl)phosphonate (BIMP) and the effects of BIMP on COX-2 expression in SK-N-SH human neuroblastoma cells. Exposure to BIMP changed cell morphology and induced caspase-dependent apoptotic cell death accompanied by cleavage of caspase 3, caspase 9, and poly (ADP-ribose) polymerase (PARP). It also increased COX-2 expression, while pretreatment with a COX inhibitor, ibuprofen, decreased BIMP-dependent cell death and COX-2 expression in SK-N-SH cells. Thus, our findings suggest that BIMP induces apoptotic cell death and upregulates COX-2 expression.

Interday accuracy and precision were 81.3−100.0% and 0.8−9.9%, respectively. The analytical values of SRM 2669 obtained by the proposed method were sufficiently accurate. Conclusions: The proposed method over- came the disadvantages of high-performance liquid chromatography with inductively coupled plasma mass spectrometry. It was a robust, selective and cost-effec- tive method suitable for routine analyses and could be useful for the biological monitoring of occupational exposure to inorganic arsenic. (J Occup Health 2012; 54: 434-440) Abstract: Development of an Analytical Method for the Determination of Arsenic in Urine by Gas Chro- matography-mass Spectrometry for Biological Monitoring of Exposure to Inorganic Arsenic: Akito TAKEUCHI, et al. Osaka Occupational Health Service Center, Japan Industrial Safety and Health Associa- tion—Objectives: The purpose of this study was to develop an analytical method for the simultaneous determination of inorganic arsenic (As(III) and As(V)) and monomethylarsonic acid (MMA) in urine by gas chromatography-mass spectrometry (GC-MS) for the biological monitoring of exposure to inorganic arsenic. Methods: Arsenic compounds (after reduction of arse- nic to the trivalent state) were derivatized with 2,3- dimercapto-1-propanol and then analyzed using a GC-MS. The proposed method was validated according to the US Food and Drug Administration guidelines. The accuracy of the proposed method was confirmed by analyzing Standard Reference Material (SRM) 2669 (National Institute of Standards and Technology). Results: Calibration curves showed linearity in the range 1−100 µg/l for each of the arsenic species, with correlation coefficients of >0.999. For each of the arse- nic species, the limits of detection and quantification were 0.2 µg/l and 1 µg/l, respectively. The recoveries were 96−100%, 99−102% and 99−112% for As(III), As(V) and MMA, respectively. Intraday accuracy and precision were 82.7−99.8% and 0.9−7.4%, respectively.

It is known that tricyclic antidepressants induce long QT intervals associated with forms of life-threatening arrhythmia such as torsades de pointes (TdP), and these adverse effects may also occur in neonates whose mothers take tricyclic antidepressants. We report a neonatal case of prolonged QT interval and TdP caused by clomipramine that was transferred transplacentally from the mother. Administration of magnesium sulfate was effective to abolish TdP.When mothers take tricyclic antidepressants during pregnancy, their newborns should be watched carefully for drug-induced long QT syndrome and TdP.•Tricyclic antidepressant can prolong the QT interval. It may be used for depression in pregnancy. What is New: •This is the first neonatal case report of prolonged QT interval and TdP caused by clomipramine transferred transplacentally from the mother.

A rapid and specific method for simultaneous quantitation of organophosphates [fenitrothion (MEP), malathion (MAL), and phenthoate (PAP)], glyphosate (GLYP), and glufosinate (GLUF) in human serum and urine by gas chromatography–mass spectrometry (GC–MS) was validated. All targeted compounds, together with the internal standards [fenitrothion-d 6 (MEP-d 6) and dl-2-amino-3-phosphonopropionic acid (APPA)], were extracted from the serum and urine with a mixed-mode C–C18 monolithic spin column. Extracted samples were derivatized with N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) containing 1% tert-butyldimethylchlorosilane (TBDMCS). Linear calibration curves for serum and urine spanned the ranges of 0.1–10.0 μg/ml for organophosphates, 10–100.0 μg/ml for GLYP, and 5–100.0 μg/ml for GLUF. Detection limits for serum and urine were 10 and 5 μg/ml for GLYP and GLUF, respectively, and quantitation limits for serum and urine were 10 and 5 μg/ml for GLYP and GLUF, respectively. Both detection and quantitation limits for serum and urine were 0.1 μg/ml for organophosphates. The intra-assay and interassay accuracy and precision (expressed as relative standard deviation, %RSD) evaluated for all compounds were within 96.5–109.5% and 5.9–13.1%, respectively. The recovery of organophosphates from serum and urine, spiked at concentrations of 0.1, 1.5, 5, and 9 μg/ml, ranged from 58.8 to 90.8%. Recovery of GLYP and GLUF from serum and urine, spiked at concentrations of 10 (serum) or 5 (urine), 15, 50, and 90 μg/ml, ranged from 2.6 to 6.9%. This method had a chromatographic run time of 16.0 min. The validated method was successfully applied to a clinical case of GLYP poisoning.

A rapid, specific, and sensitive method for the simultaneous quantitation of organophosphates (fenitrothion (MEP), malathion, and phenthoate (PAP)), glufosinate (GLUF), and glyphosate (GLYP) in human serum and urine by gas chromatography-mass spectrometry (GC-MS) has been validated. All of the targeted compounds together with the internal standard were extracted from the serum and urine using a mix-mode TiO-C(18) monolithic spin column. The recovery of organophosphates from serum and urine ranged from 12.7 to 49.5%. The recovery of GLUF and GLYP from serum and urine ranged from 1.9 to 7.9%. The intra- and inter-accuracy and precision (expressed as relative standard deviation, %RSD) were within 96.7-107.7% and 4.0-13.8%, respectively. The detection and quantitation limits for serum and urine were 0.1 and 0.1 µg/ml, respectively, for organophosphates, 0.1 and 0.5 µg/ml, respectively for GLUF and GLYP. The method had linear calibration curves ranging from 0.1 to 25.0 µg/ml for organophosphates and 0.5-100.0 µg/ml for GLUF, and GLYP. The validated method was successfully applied to a clinical GLYP poisoning case.

A method coupling spin column extraction with gas chromatography-mass spectrometry was developed for the simultaneous extraction of acidic and basic drugs from urine. Benzodiazepines, local anaesthetics, antidepressants, and barbiturates were used as model drugs. Sample loading, washing, and elution of the target drugs were accomplished by centrifugation of the column. In this study, mixed-mode monolithic silica bonded with a C18 reversed-phase and a strong cation exchange phase was packed in a spin column. The pH of a urine sample (0.2 mL) was adjusted to 3 and the analytes adsorbed onto the column were eluted with 0.1 mL of MeOH containing 2% NH3; all the tested drugs were simultaneously extracted from urine. The recovery of the tested drugs was 65-123%. Up to a concentration of 2500 ng/mL of the target drugs in urine, a linear curve was observed (r(2)>0.996). The intra- and interday RSDs at three different concentrations in urine were 2.1-14.7%. For RSDs lower than 15%, the limits of detection were 1-25 ng/mL. The proposed method was successfully applied for clinical and forensic cases and the results thus obtained were in good agreement with those obtained by conventional methods.

Monolith was first used as a material for chromatographic separation two decades ago and solid-phase extraction over 10 years, and since then, separation science has undergone a dramatic change owing to advancements in analytical technology. Recently, monolith has been modified to suit various devices for the extraction and enrichment of analytes in any matrices of environmental, food, and biological analyses. This approach has contributed to miniaturization and automation for sample preparation, and it can reduce the time and cost requirements of sample preparation. Recently, numerous applications have been demonstrated for online and inline preconcentration coupled with monolith, and many kinds of devices have been designed and developed for offline devices. In this review, these applications and devices are listed and discussed in reference to other fields.

分析技術の進歩に伴い，医薬，臨床など多くの分野で微量分析が行われている．微量分析を可能にするには，分析機器の高感度化，分析方法の最適化，試料の前処理における濃縮は重要で，更なる効率的で確かな方法が求められている．著者らは，シリカモノリスへ強カチオン交換基と強アニオン交換基を化学結合させたモノリススピンカラムを開発し，生体試料中からのイオン性薬物の固相抽出法へ応用した．シリカモノリスは支持体と抽出相が一体型のため，スピンカラムに成形する際，固相抽出カラムのようにフィルターで挟み込む必要が無く，厚さもわずか数mmと極めて薄い．更に，遠心機で抽出を行うため同時に多数の検体処理が可能となったほか，抽出溶媒も少量で行え濃縮操作も省略可能となり固相抽出より抽出操作の簡略化が可能となった．イオン交換型シリカモノリススピンカラムを用いた固相抽出法は，生体試料中のイオン性化合物の抽出に有効であることが示された．

A rapid gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the analysis of eperisone in serum using monolithic spin-column extraction. The linear concentration range for eperisone was 2-2500 ng/ml. The limit of detection was found to be 0.5 ng/ml. The average extraction recovery range was 92.8-96.0%. The intra- and interday relative standard deviations (RSDs) of the concentrations were less than 12.6% and 12.5%, respectively. The accuracy of this method ranged from 95.0% to 98.3%. We successfully used this assay to analyze serum samples from an eperisone-overdose patient. Our method has some analytical advantages compared with previously reported gas chromatography (GC) and GC-MS methods, such as higher selectivity and sensitivity than GC with nitrogen-phosphorus detection and the avoidance of nonspecificity, the ability to use a smaller sample volume than that required for the GC-MS method, and a shorter sample preparation time than the previous solid-phase extraction (SPE) method.

We describe and validate a gas chromatography-mass spectrometry (GC-MS) method for the simultaneous quantitative detection of nine cold medication compounds and bromoisovaleryl urea an over-the-counter cold medication, in human serum; the nine compounds are acetaminophen (APAP), codeine, dihydrocodeine, three ephedrines, ethenzamide, ibuprofen, and salicylic acid. After adding the internal standard, acetaminophen-d 4 (APAP-d 4), compounds were extracted from the serum samples in the monolithic spin column. The extracted samples were evaporated to dryness. The residues were derivatized with acetonitrile and N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide + 1% tert-butyldimethylchlorosilane, which were subjected to GC-MS analysis. The limit of quantification (LOQ) was 0.005–0.1 μg mL−1. The calibration curves were linear (r 2 > 0.995) in the concentration range from the LOQ to 10 μg mL−1. The intra- and inter-day variations, determined by the measurement of quality control samples at three tested concentrations, showed acceptable values. The lower limit of detection was between 0.005 and 0.05 μg mL−1. Mean recoveries from the serum samples were between 2.5 and 73.8%. This procedure was applied in the toxicological analysis of an intoxicated patient who was responsible for a traffic accident.

To overcome the limitations of solid-phase extraction, we developed a device comprising a spin column packed with octadecyl silane-bonded monolithic silica for extracting amphetamines and methylenedioxyamphetamines from urine. Urine (0.5mL), buffer (0.4mL), and methoxyphenamine (internal standard) were directly put into the preactivated column. The column was centrifuged (3000rpm, 5min) for sample loading and washed. The adsorbed analytes were eluted and analyzed by high-performance liquid chromatography, without evaporation. The results were as follows: linear curves (drug concentrations of 0.2-20microg/mL); correlation coefficients >0.99; detection limit, 0.1microg/mL. The proposed method is not only useful for drugs from biological materials but also highly reproducible for the analysis of these drugs in urine.

A simple, sensitive, and specific liquid chromatography–mass spectrometry (LC–MS) method for simultaneous determination of dibucaine and naphazoline from serum was developed and validated. The extraction procedure was performed using a monolithic silica spin column. Chromatographic separation of dibucaine and naphazoline was achieved on a C 18 reverse phase column with a mobile phase gradient (mobile phase A: 10 mM ammonium formate and mobile phase B: acetonitrile) at a flow rate of 0.2 mL/min. LC–MS was operated under the selective ion monitoring mode using the electrospray ionization technique in the positive mode. The retention times for naphazoline, dibucaine, and the internal standard (IS) were 6.7, 7.8, and 8.0 min, respectively. A linear graph was obtained for dibucaine and naphazoline with correlation coefficients >0.998 for all analytes by this method. The limit of quantification of dibucaine and naphazoline was 10 and 25 ng/mL, respectively. The mean recoveries were greater than 70%. Both compounds were stable under conditions of short-term storage, long-term storage as well as after freeze–thaw cycles. Monolithic spin column extraction and LC–MS analysis enabled the separation of dibucaine and naphazoline within 20 min.