Evaluation of Brodifacoum-induced Toxicity by Metabonomics Approach Based on HPLC-TOF-MS / 法医学杂志

OBJECTIVES@#To analyse the metabolic changes in urine of rats with brodifacoum intoxication, and to reveal the molecular mechanism of brodifacoum-induced toxicity on rats.@*METHODS@#By establishing a brodifacoum poisoning rats model, the urine metabolic profiling data of rats were acquired using high performance liquid chromatography-time of flight mass spectrometry （HPLC-TOF-MS）. The orthogonal partial least squares analysis-discrimination analysis （OPLS-DA） was applied for the multivariate statistics and the discovery of differential metabolites closely related to toxicity of brodifacoum.@*RESULTS@#OPLS-DA score plot showed that the urinary metabolic at different time points before and after drug administration had good similarity within time period and presented clustering phenomenon. Comparing the urine samples of rats before drug administration with which after drug administration, twenty-two metabolites related to brodifacoum-induced toxicity were selected.@*CONCLUSIONS@#The toxic effect of brodifacoum worked by disturbing the metabolic pathways in rats such as tricarboxylic cycle, glycolysis, sphingolipid metabolism and tryptophan metabolism, and the toxicity of brodifacoum is characterized of accumulation effect. The metabonomic method based on urine HPLC-TOF-MS can provide a novel insight into the study on molecular mechanism of brodifacoum-induced toxicity.

Identification of Methamphetamine Abuse and Selegiline Use: Chiral Analysis of Methamphetamine and Amphetamine in Urine / 法医学杂志

Objective To study the content variation of selegiline and its metabolites in urine, and based on actual cases, to explore the feasibility for the identification of methamphetamine abuse and selegiline use by chiral analysis. Methods The urine samples were tested by chiral separation and LC-MS/MS method using CHIROBIOTICTM V2 chiral liquid chromatography column. The chiral analysis of metham-phetamine and amphetamine were performed on the urine samples from volunteers of selegiline use and drug addicts whom suspected taking selegiline. Results After 5 mg oral administration, the positive test time of selegiline in urine was less than 7 h. The mass concentrations of R(-)-methamphetamine and R(-)-amphetamine in urine peaked at 7 h which were 0.86μg/mL and 0.18μg/mL and couldn't be de-tected after 80 h and 168 h, respectively. The sources of methamphetamine and amphetamine in the urine from the drug addicts whom suspected taking selegiline were analysed successfully by present method. Conclusion The chiral analysis of methamphetamine and amphetamine, and the determination of selegi-line's metabolites can be used to distinguish methamphetamine abuse from selegiline use.

Biomonitoring of 33 Elements in Blood and Urine Samples from Coastal Populations in Sanmen County of Zhejiang Province / 法医学杂志

OBJECTIVE@#To determine the normal reference values of 33 elements, Ag, Al, As, Au, B, Ba, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, Hg, Li, Mg, Mn, Mo, Ni, Pb, Rb, Sb, Se, Sr, Th, Ti, Tl, U, V, Zn and Zr, in the blood and urine samples from the general population in Sanmen County of Zhejiang province, a typical coastal area of eastern China.@*METHODS@#The 33 elements in 272 blood and 300 urine samples were determined by inductively coupled plasma-mass spectrometry (ICP-MS). The normality test of data was conducted using SPSS 17.0 Statistics. The data was compared with other reports.@*RESULTS@#The normal reference values of the 33 elements in the blood and urine samples from the general population in Sanmen County were obtained, which of some elements were found to be similar with other reports, such as Co, Cu, Mn and Sr, while As, Cd, Hg and Pb were generally found to be higher than those previously reported. There was a wide variation between the reports from different countries in blood Ba.@*CONCLUSION@#The normal reference values of the 33 elements in the blood and urine samples from the general population in Sanmen County are established, and successfully applied to two poisoning cases.

Chlorpyrifos Determined in Human Blood by UPLC-MS/MS and Its Application in Poisoning Cases / 法医学杂志

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.

Normal Range of 33 Elements in Blood Samples from General Population in Hunan Province / 法医学杂志

OBJECTIVE@#To determine the normal range of the 33 elements (Li, Be, B, Mg, Al, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Zr, Mo, Ag, Cd, Sb, Cs, Ba Au, Hg, Tl, Pb, Th and U) in human whole blood of general population in Hunan province.@*METHODS@#Blood samples were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) to determine the normal range. The influences of district, gender and age to the element content in blood samples were also observed.@*RESULTS@#The normal range of 33 elements in blood samples from general population in Hunan province were obtained. Gender was shown to statistically influence the concentrations of B, Mg, Ca, Ti, Mn, Fe, Co, Cu, Zn, As, Se, Rb, Sr, Ag, Cd, Cs, Hg and Pb (P < 0.05), while age was shown to influence the concentrations of Co, Ni, Cs and Hg in women (P < 0.05) as well as Cu, Se and Hg in men (P < 0.05).@*CONCLUSION@#Although there are variables in different districts, the normal ranges of trace element in blood of the four cities in Hunan province are established.

Method development and validation for determining 1,3-butadiene in human blood by gas chromatography-mass spectrometry and head-space gas chromatography / 法医学杂志

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.

Determination of 33 inorganic elements in human hair by electricity plate digestion and inductively coupled plasma-mass spectrometry / 法医学杂志

OBJECTIVE@#To establish the electricity plate digestion and inductively coupled plasma-mass spectrometry (ICP-MS) method for determination of 33 inorganic elements in human hair.@*METHODS@#Lithium (6Li), Germanium (72Ge), Yttrium (89Y), Indium (115In), and Terbium (159Tb) were used as internal standards. The electric heating board digestion in a mixture of nitric acid and hydrogen peroxide was used as the pre-treatment of the hair. Thirty-three inorganic elements in human hair were analyzed by ICP-MS method.@*RESULTS@#The detection limit of ICP-MS was 0.0001 microg/g(Th)-10.9 microg/g (Ca) and the limit of quantitation was 0.0005 microg/g (Th)-25 microg/g (Ca). The recovery rate of this method was 86%-113%. The RSD for the intra-day and inter-day were less than 9.2%. The method was not statistically different from microwave digestion method.@*CONCLUSION@#This method is highly efficient and accurate. It can be used for analysis of 33 inorganic elements in human hair.

Screening and confirmation of psychotropic drugs in blood and urine by HPLC-LTQ Orbitrap MS / 法医学杂志

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.

Measurement of acetonitrile in blood and urine by head-space gas chromatography / 法医学杂志

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.

Developments in determination of elements using ICP-MS in blood and urine / 法医学杂志

Inductively coupled plasma-mass spectrometry (ICP-MS) is the most common technique for elements analysis at present. ICP-MS with high sensitivity and wide linear range can be applied to multi-elements analysis in blood and urine. This paper reviews the common means of sample pretreatment (direct dilution method and wet digestion method), the method for correction of mass spectral interference and non-interference, the main influence factors of analysis results, and provides an outlook of the application of ICP-MS in forensic toxicological analysis.

Simultaneous determination of sixteen antibiotics in human urine with ultra performance liquid chromatography-tandem mass spectrometry / 法医学杂志

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.

Determination of Hg in biological samples by inductively coupled plasma mass spectrometry / 法医学杂志

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.

Application of liquid chromatography-high resolution mass spectrometry in toxicological screening / 法医学杂志

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 24 elements in human hair by ICP-MS using microwave digestion / 法医学杂志

OBJECTIVE@#To establish an inductively coupled plasma-mass spectrometry (ICP-MS) method for determination of 24 elements in human hair.@*METHODS@#The samples were digested with microwave digestion instrument. ICP-MS was applied to determine 24 elements in human hair using indium (115In) as an internal standard. The established method was applied to determine element concentration in normal group (56 samples) and heroin abuse group (10 samples).@*RESULTS@#The limits of detection ranged from 0.000 3 microg/g to 10.14 microg/g. Measured value of the standard materials were basically consistent with the standard value. The contents of magnesium, gallium and barium in hair of heroin addicts decreased after rehabilitation treatment.@*CONCLUSION@#The method is sensitive and accurate for determination of 24 elements in human hair.

The influence of estazolam on human neurobehavioral function / 法医学杂志

OBJECTIVE@#To study the relation between human blood estazolam concentration and neurobehavioral function.@*METHODS@#The neurobehavioral ability of 10 volunteers were measured with computer-administered neurobehavioral evaluation system-chinese3 (NES-C3) and SMART EquiTest system.@*RESULTS@#The neurobehavioral ability and balance function declined 1 h later after dosing estazolam. The neurobehavioral ability index and balance function declined to the lowest level 3 h later after dosing estazolam. The neurobehavioral ability recovered partly 6 h later after dosing estazolam, and neurobehavioral ability recovered completely 10 h later.@*CONCLUSION@#Driving ability was impaired when estazolam concentration in blood is 20 ng/mL, and the neurobehavioral ability declined when estazolam concentration is 40 ng/mL in blood. The influence to human in absorption process is greater than the metabolic process with the same estazolam concentration.

Simultaneous determination of opioid compounds in human urine by UPLC-MS/MS / 法医学杂志

OBJECTIVE@#To propose a method for simultaneous determination of codeine(COD), 6-monoacetyl-morphine (6-MAM), morphine (MOR), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in human urine by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS).@*METHODS@#After precipitation of protein by acetonitrile, the urine samples, with added the morphine-d3 (MOR-d3) and morphine-3-Glucuronide-d3 (M3G-d3) as internal standards, were pre-treated by Sirocco protein precipitation plate, and then analyzed by UPLC-MS/MS.@*RESULTS@#The limit of detection was 0.2 ng/mL for both COD and MAM, the limit of quantitation was 0.5 ng/mL for both COD and MAM. The limit of detection was 0.5 ng/mL for MOR, M3G and M6G, the limit of quantitation was 1 ng/mL for them. The linear correlation coefficients were not less than 0.9997, both the inter-day and intra-day precisions were less than 10%, the recoveries were in the range of 70.0% to 98.3%, the matrix effects were about 50.5% to 99.0%.@*CONCLUSION@#This proposed method is simple, rapid and accurate, it could be applied in forensic toxicological analysis.

Determination of ethyl glucuronide in blood and urine by LC-MS/MS / 法医学杂志

OBJECTIVE@#To develop a method for determining ethyl glucuronide (EtG) in blood and urine by liquid chromatograph coupled with tandem mass spectrometer (LC-MS/MS).@*METHODS@#After blood and urine de-proteined by acetonitrile, the supernate obtained from a centrifuge was analyzed by LC-MS/MS.@*RESULTS@#Determination limit of EtG in both blood and urine was 0.05 pg/mL, with a linear range of 0.10-5.00 microg/mL (r > 0.999). Accuracy in both matrixes was 95%-109%. Inter- and intra-day RSD were less than 12%. The method showed an excellent performance when it was used to analyze authentic blood and urine samples for EtG.@*CONCLUSION@#The method is capable for blood and urine EtG analysis.

Determination of Cr, Cd, As, Tl and Pb in blood by inductively coupled plasma mass spectrometry / 法医学杂志

OBJECTIVE@#To establish an inductively coupled plasma mass spectrometry (ICP-MS) method for determination of Cr, Cd, As, Tl and Pb in blood.@*METHODS@#The samples were digested by microwave digestion instrument. ICP-MS was applied to determine Cr, Cd, As, Tl and Pb in blood by using 115In as an internal standard.@*RESULTS@#The limits of detection were in the range of 0.00001-0.00249 microg/L. The accuracy of the method ranged from 90.1% to 110.7% and the precision ranged from 4.0% to 7.9%.@*CONCLUSION@#The method is accurate and rapid with superior sensitivity and linear range. It could be used in the poisoning cases caused by Cr, Cd, As, Tl and Pb.

Detection and application of ethyl glucuronide in forensic toxicology / 法医学杂志

Ethyl glucuronide is a specific metabolite of ethanol. There have been plenty of articles referring its pharmacokinetics, detection and application as a specific bio-marker of alcohol intake. This article reviews various analytical methods of EtG, relationship between EtG quantification and ethanol intake, and criteria for determining chronic alcohol abuse, and origin of ethanol found in the cadavers by EtG analysis. EtG has its potential application in forensic toxicology.

Determination of ethyl glucuronide in blood by GC-MS/MS / 法医学杂志

OBJECTIVE@#To develop a method for determining ethyl glucuronide(EtG) in human blood with gas chromatograph-tandem mass spectrometry (GC-MS/MS).@*METHODS@#Human blood protein was precipitated with acetonitrile. The supernatant was transferred and air flow dried after centrifugated. The residue was derived with N, O-bis (trimethylsilyl)trifluoroacetamide (BSTFA), and analyzed with GC-MS/MS.@*RESULTS@#The detection limit of EtG in blood was 0.05 microg/mL. Calibration curve covered a span from 0.1-10 microg/mL with a good linear relationship (r = 0.999 9). The method showed a excellent performance when was used to authentic blood sample analysis for EtG.@*CONCLUSION@#The method is suitable for blood EtG analysis.