Detection and Analysis of Blood Dexmedetomidine in Drug-Facilitated Cases.

OBJECTIVES: To establish a simple and rapid qualitative and quantitative detection method of dexmedetomidine in blood. METHODS: Blood was separated on the Allure PFP Propyl liquid chromatography column with isocratic elution after it was precipitated by acetonitrile and filtered. Qualitative and quantitative analysis of dexmedetomidine was performed using positive ion scan mode and multi-reaction monitoring mode. RESULTS: The limit of detection of dexmedetomidine in blood was 0.2 ng/mL and the limit of quantification was 0.5 ng/mL. The linearity of the method was good in the range of 0.5-1 000 ng/mL, and the correlation coefficient was greater than 0.99. The accuracy of the method was 90.34%-112.67% and the extraction recovery was 50.05%-91.08%, with no significant matrix effect. CONCLUSIONS: This method is simple, selective and suitable for the qualitative and quantitative analysis of dexmedetomidine in blood, which can provide a reference for drug-facilitated cases involving dexmedetomidine.

Strategies for Collection and Analysis of Samples in Simple Asphyxiant Gas Acute Poisoning Death Cases.

At present, the death cases of simple asphyxiant gas acute poisoning are increasing sharply. Common asphyxiant gases in death cases include nitrogen, helium, carbon dioxide, methane, propane, laughing gas, etc. Simple asphyxiant gas has no affinity for biological matrices and escapes quickly, which puts forward new requirements for autopsy procedures, selection and collection of samples, laboratory analysis and identification. This paper reviews the research and development process of death cases caused by simple asphyxiant gas acute poisoning and put forwards the collection and analysis strategy of the samples in such cases. The most valuable biological samples in such cases should be lung tissues associated with the airways, followed by brain tissue and cardiac blood. Gaseous samples from the esophageal cavity, tracheal cavity, pulmonary bronchi, gastric and cardiac areas are also recommended as valuable samples. In the case of postmortem examination, the gas should be injected into gas sample bag directly. Biological materials such as tissue and blood should be directly sealed in head-space vials and analyzed by using the headspace gas chromatography-mass spectrometry.

Analysis of the Distribution of Total Phosphine and the Characteristics of Phosphine Poisoning in 29 Victims.

OBJECTIVES: To study the distribution of total phosphine in phosphine poisoning victims and summarize the characteristics of phosphine poisoning cases. METHODS: The phosphine and its metabolites in the biological samples of 29 victims in 16 phosphine poisoning cases were qualified and quantified by headspace gas chromatography-mass spectrometry. RESULTS: Five victims among 29 were poisoned by ingestion of aluminium phosphide and 24 by inhalation of phosphine gas. Phosphine metabolites were detected in the biological samples of 23 victims, and the concentrations of total phosphine in blood ranged 0.5-34.0 µg/mL. The total concentration of phosphine in liver tissue was up to 71.0 µg/g. Phosphine was not detected in the blood of the other six survived victims, which may be related to the small amount of phosphine exposure and the delay in blood sampling. CONCLUSIONS: The total concentration of phosphine in blood and tissues caused by aluminum phosphine ingestion is higher than that caused by phosphine gas inhalation. The death cases of phosphine inhalation are characterized by long exposure time, repeated exposures and age susceptibility.

Forensic Analysis of 9 Poisoning Death Cases Caused by Oral Administration of Diphenidol.

OBJECTIVES: To analyze the characteristics of diphenidol poisoning cases and to provide clues and technical means for the identification of such cases. METHODS: Biological samples of 9 deaths caused by diphenidol poisoning were detected by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and the characteristics of these cases were analyzed retrospectively. RESULTS: Most of the deaths caused by diphenidol poisoning were young females. The dosage was between 60 and 300 tablets, and the mass concentration of diphenidol in the postmortem blood ranged from 0.87 to 99.00 µg/mL. There was no correlation between the dosage and the concentration of diphenidol in the blood. CONCLUSIONS: Diphenidol poisoning has the characteristics of high concealment and lethality. More attention should be paid to suicide cases, and diphenidol should be recommended as a routine detection item to avoid missing detection.

Simultaneous determination of 11 illicit drugs and metabolites in wastewater by UPLC-MS/MS.

Assessing collective drug consumption based on the concentrations of illicit drugs and their metabolites in wastewater is a new technology. Currently, this technology is receiving attention in China, and methods for multiple illicit drug detection in wastewater are urgently needed. In our study, a method with a short runtime (7 min), a small solid-phase extraction (SPE) loading volume (50 mL) and high sensitivity (lower limits of quantitation (LLOQs) ranged from 0.2 to 5 ng/L) was developed for the simultaneous determination of amphetamines, ketamine, opiates, cocaine and their metabolites in wastewater. Samples were enriched by SPE on a mixed-mode sorbent (Oasis MCX) and analyzed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The limits of detection (LODs) ranged from 0.1 to 2 ng/L, and the LLOQs varied between 0.2 and 5 ng/L. Moreover, the method developed was applied to real wastewater samples collected from 15 different wastewater treatment plants (WWTPs). In the results, the most abundant compounds were morphine (1.8-46.6 ng/L) and codeine (3.7-24.9 ng/L), which were detected in 13 WWTPs. After successful optimization of the UPLC-MS/MS conditions and sample loading pH, the method developed is able to meet the needs of common illicit drug monitoring and high-throughput analysis requirements.

Diagnosis of aluminum phosphide poisoning using a new analytical approach: forensic application to a lethal intoxication.

Aluminum phosphide (AlP) is an effective and cheap pesticide that is commonly used worldwide, but it is also a common cause of human poisoning and carries a high mortality rate. AlP reacts with moisture in air, water, and hydrochloric acid in the stomach to produce phosphine (PH3) gas. Two routes of exposure are ingestion of AlP and inhalation of phosphine generated by the action of moisture on AlP. Absorbed phosphine is rapidly metabolized into phosphite and hypophosphite. A method is described for the analysis of the phosphine metabolites in various biological matrices. The method involves reacting the sample with zinc and aqueous H2SO4 in a volatile organic analysis vial. The metabolites were transformed into phosphine gas and then analyzed by headspace gas chromatography coupled with mass spectrometry (HS-GC-MS). This method is capable of detecting quantities of PH3 as low as 0.2 µg/mL in a sample. After validation, the method was applied to animal experiments and a real case of human AlP intoxication. This approach has the advantage of detecting metabolites of PH3, in case the PH3 was converted, and can be considered a useful additional tool for the diagnosis of AlP poisoning in forensic science.

[Analysis of GHB and Its Precursors in Urine and Their Forensic Application].

OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 µg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION: The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.

Chlorpyrifos Determined in Human Blood by UPLC-MS/MS and Its Application in Poisoning Cases.

OBJECTIVE: To determine the chlorpyrifos in human blood by liquid chromatography-tandem mass spectrometry and to validate its application in poisoning cases. METHODS: The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 MGII column (250 mm x 2.0 mm, 5 µm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammonium acetate) and solvent B (methanol with 2 mmol/L ammonium acetate) at 5:95 V:V). RESULTS: The linear ranged from 5 to 500 ng/mL (r = 0.998 7). The limit of detection (LOD) and the lower limit of quantification (LLOQ) were 2 ng/mL and 4 ng/mL, respectively. For this method, the precision and accuracy of intra-day and inter-day were < 10% and 97.44%-101.10%, respectively. The results in stability test of long-term frozen were satisfied. The matrix effect, recovery and process efficiency were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. CONCLUSION: This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.

Mass imaging of ketamine in a single scalp hair by MALDI-FTMS.

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) coupled with mass spectrometry imaging (MSI) is a rapidly emerging technology that produces distribution maps of small pharmaceutical molecules in situ in tissue sections. Segmental hair analysis provides useful information regarding the state and history of drug use. A preliminary MALDI-Fourier transform ion cyclotron resonance (FTICR)-MSI method was developed for direct identification and imaging of ketamine in hair samples. After decontamination, the scalp hair samples from ketamine users were scraped gently and were fixed onto a stainless steel MALDI plate using double-sided adhesive tape. A Bruker 9.4 T solariX FTICR mass spectrometer with continuous accumulation of selected ions function was used in the positive ion mode. Four single hairs from the same drug abuser were analyzed. Three of four single hairs demonstrated ketamine spatial distribution, while only traces of ketamine were identified in the other one. The platform could provide detection power of ketamine down to the 7.7 ng/mg level in hair. MALDI-FTICR-MSI demonstrated the drug distribution over the whole hair length with higher spatial resolution compared with the traditional LC-MS/MS method after scissor cutting. Greater caution is needed in the interpretation of a single hair result because of the considerable variations in the growth rate and sample collection.

Determination of bromadiolone and brodifacoum in human hair by liquid chromatography/tandem mass spectrometry and its application to poisoning cases.

RATIONALE: Bromadiolone and brodifacoum, two common anticoagulant rodenticides, are involved in the majority of anticoagulant rodenticide poisoning cases in humans in China. Hair analysis can provide long-term information on drug exposure. A method using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) was developed and validated for the measurement of bromadiolone and brodifacoum in human hair. METHODS: A 1 mL aliquot of a phosphate buffer solution (pH 6.8) was added to 20 mg of pulverized hair followed by ultrasonication and liquid-liquid extraction. Liquid chromatography was performed using a C(18) column with a mobile phase gradient of ammonium acetate (10 mM) and methanol. A tandem mass spectrometer in multiple reaction monitoring mode with a negative electrospray ionization source was employed for detection. Warfarin-d(5) was used as an internal standard for both analytes. RESULTS: The limits of detection (LODs) for bromadiolone and brodifacoum were 0.010 and 0.025 ng/mg, respectively. The calibration curves for both analytes were linear from 0.025 to 1 ng/mg. The accuracy ranged from 90.3 to 109.3%, and the intra-day and inter-day imprecisions were less than 15%. CONCLUSIONS: The established method was found effective when applied to the analyses of bromadiolone or brodifacoum in five cases, indicating that segmental hair analysis could be useful for clinical and forensic purposes by identifying the time of ingestion.

Method development and validation for determining 1,3-butadiene in human blood by gas chromatography-mass spectrometry and head-space gas chromatography.

To develop a simple, validated method for identifying and quantifying 1,3-butadiene (BD) in human blood by gas chromatography-mass spectrometry (GC-MS) and head-space gas chromatography (HS-GC). BD was identified by GC-MS and HS-GC, and quantified by HS-GC. The method showed that BD had a good linearity from 50 to 500 microg/mL (r > 0.99). The limits of detection and quantification were 10 microg/mL and 50 microg/mL, respectively. Both the intra-day precision and inter-day precision were < 6.08%, and the accuracy was 96.98%-103.81%. The method was applied to an actual case, and the concentration of BD in the case was 242 microg/mL in human blood. This simple method is found to be useful for the routine forensic analysis of acute exposure to BD.

Discrimination of Traditional Chinese Medicine Syndromes in Type 2 Diabetic Patients Based on Metabolomics-Proteomics Profiles.

Materials and Methods: The metabolomics-proteomics of sixty patients with T2DM were acquired by high-performance liquid chromatography (HPLC). In addition, some clinical features, containing total cholesterol (TC), triglycerides (TG), hemoglobin A1c (HbA1c), body mass index (BMI), and low-density lipoprotein (LDL) together with high-density lipoprotein (HDL), were determined via clinical detection strategies. Abundant metabolites and proteins, respectively, were identified with the analysis of liquid chromatography tandem mass spectrometry (LC-MS/MS). Results: 22 differentially abundant metabolites and 15 differentially abundant proteins were determined. The analysis of bioinformatics suggested that the differentially abundant proteins were commonly associated with the renin-angiotensin system, vitamin digestion and absorption, hypertrophic cardiomyopathy, and so on. Furthermore, differentially abundant metabolites were amino acids and were associated with the biosynthesis of CoA and pantothenate, together with the metabolisms of phenylalanine, beta-alanine, proline, and arginine. Combination analysis revealed that the vitamin metabolism pathway was predominantly affected. Conclusions: DHS syndrome can be separated by certain metabolic-proteomic differences, and metabolism is particularly prominent, especially in vitamin digestion and absorption. From the molecular level, we provide preliminary data for the extensive application of TCM in the study of T2DM, and at the same time benefited in a sense diagnosis and treatment of T2DM.

[Measurement of acetonitrile in blood and urine by head-space gas chromatography].

OBJECTIVE: To establish the method for measurement of acetonitrile in blood and urine by head-space gas chromatography. METHODS: DB-ALC1 (30 m x 320 microm x 1.8 microm) and DB-ALC2 (30 m x 320 microm x 1.2 microm) capillary column were used to measure the acetonitrile in blood and urine with the isopropanol as internal standard reference. RESULTS: The limits of detection of acetonitrile in both blood and urine were 0.5 microg/mL, with a linear range of 5-1000 microg/mL (r = 0.999).The accuracy of this method was 93.2%-98.0%. The RSD for the intra-day and inter-day were less than 3.7%. CONCLUSION: The method is capable for measurement analysis of acetonitrile in blood and urine.

[Screening and confirmation of psychotropic drugs in blood and urine by HPLC-LTQ Orbitrap MS].

OBJECTIVE: To establish a screening and confirmation method for psychotropic drugs and their metabolites in human blood and urine by HPLC-LTQ Orbitrap MS. METHODS: The samples were pretreated with Sirocco protein precipitation plate, and then analyzed by HPLC-LTQ Orbitrap MS. The method was validated in terms of the limit of detection (LOD). An accurate mass database was created for psychotropic drugs screening. RESULTS: The LOD for most of 56 determined compounds was < or = 0.1 ng/mL. The accurate mass database included the accurate mass information of 61 psychotropic drugs. CONCLUSION: The method is accurate, rapid, sensitive and the database is suitable for psychotropic drugs screening and confirmation.

Disposition of ketamine and norketamine in hair after a single dose.

As a rapid-acting dissociative anesthetic, ketamine has been used in drug-facilitated crimes. The aim of this study is to investigate the disposition of ketamine and its main metabolite norketamine in hair after a single dose of ketamine. Four healthy volunteers were recruited into the study. Hair was collected 1, 2, 3, 4, 8, 12 and 16 weeks after a single oral dose of ketamine solution (10 mg) and analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry. The wet cotton swab wiped the scalp of the subjects at 1 h, 24 h, 48 h and 1 week after administration. Maximum hair concentrations (C (max)) for ketamine and norketamine were 19.0 ± 6.5 and 18.7 ± 13.3 pg/mg, respectively. Except for the first week, the ratio of ketamine to norketamine in most of segments (87.5%) was greater than 1. All the cotton swab samples collected at 24 and 48 h were positive. The results from cotton swabs and the concentrations of ketamine and norketamine in hair segments collected at different times showed that some of ketamine and norketamine incorporated into hair originated from sweat and sebum on the scalp of the subjects.

[Simultaneous determination of sixteen antibiotics in human urine with ultra performance liquid chromatography-tandem mass spectrometry].

OBJECTIVE: To develop a method for simultaneous determination of sixteen antibiotics in human urine by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). METHODS: With Piperacillin as an internal standard, the target antibiotics in urine samples were enriched and purified by Oasis HLB solid phase extraction (SPE) cartridges, then separated in a ZORBAX SB-C18 column with a gradient elution of mobile phase of 0.1% formic acid water and acetonitrile, finally analyzed with multiple reaction monitoring (MRM) mode. RESULTS: The limits of detection (LOD) for these sixteen antibiotics were in the range of 0.05-10.0 ng/mL and the limits of quantification (LOQ) in the range of 0.25-20.0 ng/mL. Within the related linear range, the related coefficient (r) of sixteen antibiotics were all more than 0.995. Accuracies for these antibiotics were ranged from 82.0%-119.3%, the within-day precision were less than 13.9%. CONCLUSION: The developed method is sensitive, specific and appropriate for the analysis of antibiotics in forensic toxicology and therapeutic drugs monitoring.

[Application of liquid chromatography-high resolution mass spectrometry in toxicological screening].

Due to the diversity of toxicologically relevant substances, the uncertainty of target compounds and the specificity of samples, toxicological screening techniques have always been valued by the forensic toxicologists. Depending on its powerful separation ability, superhigh resolution and accurate mass measurement, combined with the two levels spectrum database matching and abundance ratio of isotope ion, the liquid chromatography-high resolution mass spectrometry (LC-HRMS) analyzers have increasingly advantage in screening and identification of chemical compound. This review focuses on the applications of LC-HRMS in screening and identification of drug-of-abuse, prescription drugs, pesticide and stimulant. The prospect of LC-HRMS in forensic toxicology analysis is also included.

[Determination of Hg in biological samples by inductively coupled plasma mass spectrometry].

OBJECTIVE: To establish an inductively coupled plasma mass spectrometry (ICP-MS) method for determination of Hg in biological samples. METHODS: The samples were digested with microwave digestion instrument. ICP-MS was applied to detect Hg in blood, urine and hair specimens by using 115In as an internal marker. The ability of gold to eliminate the memory effect of mercury was investigated with the gold amalgamate produced by gold and mercury. RESULTS: The limits of detection were in the 0.01 microg/L, and the accuracy of the method ranged from 97.0% to 107.1%. The concentration of gold was 10 microg/L and the memory effect of mercury was resolved. CONCLUSION: The method is accurate, rapid, sensitive and suitable for the cases of mercury poisoning and the clinical diagnosis and monitoring for patients with mercury poisoning.

Determination of twenty herbicides in blood by ultrapressure liquid chromatography-tandem mass spectrometry.

A sensitive and rapid method for the simultaneous determination of twenty herbicides (aclonifen, lactofen, terbutryn, butylate, carbetamide, fluazifop-P-butyl, propanil, prometryn, isoproturon, terbumeton, pretilachlor, pendimethalin, cycloxydim, tri-allate, metolachlor, diuron, alloxydim, prosulfuron, triflusulfuron-methyl, and acetochlor) in human blood is reported herein. Liquid-liquid extraction coupled with ultra-pressure liquid chromatography-tandem mass spectrometry was employed for the simultaneous analysis of all compounds in 15 min. Validation parameters were studied through the estimation of the limits of detection and quantification, calibration curves, sensitivity, spiked recovery and precision. The limits of detection ranged from 0.1 to 1.0 ng/mL. The limits of quantification ranged from 0.5 to 2.0 ng/mL. Good linearity was obtained for all compounds with R2> 0.99 in all cases. Furthermore, interday precision (< 15%) and intraday precision (< 15%) were shown to be satisfactory. Recoveries in spiked blood samples were evaluated, and acceptable values (88.0%~108.8%) were found. Finally, this method was successfully applied to the determination of fluazifop-P-butyl, isoproturon and acetochlor in blood samples from real forensic cases. These results suggest that this method is reliable for rapid forensic and clinical diagnosis.

Metabolism of 4F-MDMB-BICA in zebrafish by liquid chromatography-high resolution mass spectrometry.

In this study, in vivo metabolic studies of the synthetic cannabinoid 4F-MDMB-BICA were investigated using zebrafish models. The metabolites were identified and structurally illustrated by liquid chromatography-high resolution mass spectrometry. Fourteen phase-I metabolites and four phase-II metabolites were generated from zebrafish. The main metabolic pathways of the phase-I metabolism included N-dealkylation, N-dealkylation combined with hydroxylation, amide hydrolysis, oxidative defluorination, oxidative defluorination to butyric acid, acetic acid formation at the indole side chain, hydroxylation, ester hydrolysis followed by hydroxylation, dehydrogenation, dehydrogenation, and N-dealkylation, and oxidative defluorination subsequently combined with dehydrogenation. The main biotransformations of the phase-II metabolism were glucuronidation and sulfation. Two phase-I metabolites (A1 and A11) and four phase-II metabolites (A2, A3, A4, and A12) were reported for the first time. A14, which was confirmed in human biological samples, was detected only in zebrafish samples but not found in human liver microsome incubation study. The current study indicates that the zebrafish model is a promising tool for elucidating the metabolism of NPS in the future.