Shigetoshi Kage

Determination of cyanide and thiocyanate in blood by gas chromatography and gas chromatography-mass spectrometry

We devised a sensitive and simple method for determining cyanide and its major metabolite, thiocyanate, in blood using an extractive alkylation technique. Pentafluorobenzyl bromide was used as the alkylating agent, and tetradecyldimethylbenzylammonium chloride was used as the phase-transfer catalyst. The derivatives obtained were analyzed qualitatively by gas chromatography-mass spectrometry and quantitatively by gas chromatography with an electron-capture detection. The detection limits of cyanide and thiocyanate were 0.01 and 0.003 mumol/ml, respectively, while the gross recovery of both compounds was 80%. The calibration curve was linear over the concentration range from 0.02 to 1.0 mumol/ml for cyanide and from 0.01 to 1.0 mumol/ml for thiocyanate. The accuracy and precision of the method were evaluated, and the coefficients of variation were found to be within 10%. Using the method, the blood levels of two victims who had died from cyanide poisoning were determined.

The usefulness of thiosulfate as an indicator of hydrogen sulfide poisoning: three cases

Abstract We examined the usefulness of thiosulfate as an indicator of hydrogen sulfide poisoning by analysing sulfide and thiosulfate in three cases. In the first (non-fatal) case sulfide and thiosulfate were not detected in the blood samples from any of the four workers involved in the accident. In the urine samples, only thiosulfate was detected in three out of the four workers at a concentration of 0.12–0.43 μmol/ml, which was 4–14 times higher than the level in a healthy person. In the second (fatal) case sulfide and thiosulfate were detected in the blood sample at concentrations of 0.007 μmol/ml for sulfide, and 0.025 μmol/ml for thiosulfate. The thiosulfate concentration was at least 8 times higher than the level in a healthy person. In the third (fatal) case sulfide and thiosulfate were detected in the blood sample at concentrations of 0.95 μmol/ml for sulfide, and 0.12 μmol/ml for thiosulfate. Based on the above results, we concluded that thiosulfate in urine is the only indicator to prove hydrogen sulfide poisoning in non-fatal cases, while the analysis of sulfide in fatal cases should be accompanied by the measurement of thiosulfate in blood.

Sulfide concentrations in postmortem mammalian tissues.

Postmortem changes in sulfide concentrations in body tissues were examined in autopsied rats exposed to hydrogen sulfide concentrations of 550 to 650 ppm, and in nonexposed rats and humans. Analyses were made by gas chromatography, following an extractive alkylation. Sulfide concentrations in the blood, liver, and kidneys of rats increased in both the exposed and nonexposed groups, depending on the lapse of time after death. On the other hand, the lung, brain, and muscle showed little or no change in sulfide concentration with elapse of time after death. The data obtained from human tissues were almost the same as those for rats, except data for blood, in which no or little increase of sulfide was observed.

Determination of nitrate in blood by gas chromatography and gas chromatography–mass spectrometry

We devised a sensitive and simple method for determining nitrate in whole blood, using an extractive alkylation technique. Nitrate in whole blood was reduced to nitrite by hydrazine sulfate in the presence of Cu2+ and Zn2+ as catalysts, and alkylated with pentafluorobenzyl bromide using tetradecyldimethylbenzylammonium chloride as the phase-transfer catalyst. The obtained derivative was analyzed qualitatively by gas chromatography-mass spectrometry and quantitatively by gas chromatography with electron-capture detection. The detection limit of nitrate in whole blood was 0.01 mM. The calibration curve was linear over the concentration range from 0.02 to 1.0 mM for nitrate in whole blood. The accuracy and precision of the method were evaluated and the relative standard deviations were found to be within 10%. Using this method, the blood nitrate levels of two victims who committed suicide by inhaling automobile exhaust gas were determined.

Fatal and nonfatal poisoning by hydrogen sulfide at an industrial waste site.

An adult man (A) entered a pit to collect seepage at an industrial waste site in Japan. As he suddenly lost consciousness, three colleagues (B, C, D) entered the pit to rescue him. All of these men lost consciousness in the pit. Two workers (A and B) died soon after the accident, one worker (C) died 22 days after the accident, and one worker (D) survived. Since hydrogen sulfide gas was detected in the atmosphere of the pit, gas poisoning was suspected. Toxicological analyses of sulfide and thiosulfate, a metabolite of sulfide, in blood and urine of the victims were made using the extractive alkylation technique combined with gas chromatography/mass spectrometry (GC/MS). Sulfide was detected in the blood of A and B at levels of 0.13 and 0.11 mg/L, respectively, somewhat higher than in healthy persons. Thiosulfate was detected in whole blood of deceased victims A and B, in the plasma of deceased victim C, at concentrations of 10.53, 4.59, and 4.14 mg/L, respectively. These values were similar to those found in fatal cases of hydrogen sulfide poisoning. Thiosulfate was not detected in the plasma of survivor D. With respect to urine samples, thiosulfate was the highest in the non-acute death victim C (137.20 mg/L), followed by that in the survivor D (29.34 mg/L), and low (0.90 mg/L) and not detected in the acute death victims, A and B, respectively. Based on these results, all four patients were victims of hydrogen sulfide poisoning. The concentrations of thiosulfate in blood and urine were more useful than that for sulfide for determining hydrogen sulfide poisoning. Thiosulfate in urine was the only indicator of hydrogen sulfide poisoning in the non-fatal victim.

A Fatal Case of Hydrogen Sulfide Poisoning in a Geothermal Power Plant

An adult man entered an oil separator room to remove waste oil from a vacuum pump in a geothermal power plant. He suddenly collapsed and died soon after. Since hydrogen sulfide gas was detected in the atmosphere at the scene of the accident, poisoning by this gas was suspected and toxicological analysis of sulfide and thiosulfate in blood, brain, lung, femoral muscle was made using the extractive alkylation technique combined with gas chromatography/mass spectrometry (GC/MS). The concentrations of sulfide in these tissues were similar to those previously reported for fatal cases of hydrogen sulfide gas. The concentration of thiosulfate in the blood was at least 48 times higher than the level in control samples. Based on these results, the cause of death was attributed to hydrogen sulfide gas poisoning.

A CASE OF DROWNING LINKED TO INGESTED SULFIDES : A REPORT WITH ANIMAL EXPERIMENTS

An adult male was found dead beneath a pool of sewage in the pump room of a fish market. Autopsy revealed the cause of death to be suffocation after aspirating sewage into the respiratory tract. Since hydrogen sulfide gas was detected in the atmosphere at the scene of the accident, gas poisoning was suspected and toxicological analysis of sulfides in body tissues was performed. The concentrations of sulfides in the blood, lung and kidney were 0.95 μmol/ml, 0.22 and 0.38 μmol/g, respectively. These values were remarkably higher than those in previously reported cases involving exposure to hydrogen sulfide gas. Therefore, oral intake of sulfides was assumed and the distribution of sulfides in tissues following oral administration of sodium sulfide solution was examined by means of animal experiments using rats. The concentration of sulfides in the blood from rats following oral intake was much higher than that seen following gas exposure. Based on these results, we concluded that the victim had been exposed to hydrogen sulfide gas and had then collapse into a pool of sewage containing sulfides. The sulfides which were distributed throughout the body tissues had mainly issued from the alimentary tract prior to death by drowning.

Determination of polysulphides in blood by gas chromatography and gas chromatography—mass spectrometry

A sensitive and simple method to determine polysulphides in human blood, using an extractive alkylation technique and gas chromatography, has been devised. Polysulphides were alkylated with pentafluorobenzyl bromide, and then converted into bis(pentafluorobenzyl)disulphide by desulphuration with potassium cyanide. The disulphide was analysed qualitatively by mass fragmentography and quantitatively by gas chromatography with electron-capture detection. The lower limit of detection was 0.005 mumol/ml. Field testing in a suicide case confirmed the validity of the method.

A Simple Method for Detection of Gunshot Residue Particles from Hands, Hair, Face, and Clothing Using Scanning Electron Microscopy/Wavelength Dispersive X-Ray (SEM/WDX)

We devised a simple and rapid method for detection of gunshot residue (GSR) particles, using scanning electron microscopy/wavelength dispersive X-ray (SEM/WDX) analysis. Experiments were done on samples containing GSR particles obtained from hands, hair, face, and clothing, using double-sided adhesive coated aluminum stubs (tape-lift method). SEM/WDX analyses for GSR were carried out in three steps: the first step was map analysis for barium (Ba) to search for GSR particles from lead styphnate primed ammunition, or tin (Sn) to search for GSR particles from mercury fulminate primed ammunition. The second step was determination of the location of GSR particles by X-ray imaging of Ba or Sn at a magnification of x 1000-2000 in the SEM, using data of map analysis, and the third step was identification of GSR particles, using WDX spectrometers. Analysis of samples from each primer of a stub took about 3 h. Practical applications were shown for utility of this method.

Determination of fluoride in human whole blood and urine by gas chromatography-mass spectrometry

We developed a simple and sensitive method for determination of fluoride in human whole blood and urine using gas chromatography-mass spectrometry (GC-MS). Fluoride was alkylated with pentafluorobenzyl bromide in a mixture of acetone and phosphate buffer (pH 6.8). The derivative obtained was analyzed by GC-MS in the positive-ion electron-impact mode. The lower limit of detection for the compound was 0.5 mg/l for both matrices. The calibration curve for fluoride was linear over the concentration range of 1–100 mg/l. The precision and accuracy of the method were evaluated, and relative standard deviation was within 10%. Using this method, levels of fluoride in whole blood and urine were determined in a case of poisoning caused by hydrofluoric acid exposure.