Rapid and sensitive quantitation of cyanide in blood and its application to fire victims.

We developed a head-space method for the determination of blood cyanide by gas chromatography with electron-capture detection. In this technique, a reaction precolumn packed with chloramine-T was used for the conversion of hydrogen cyanide into cyanogen chloride. Since the reaction precolumn eliminated the necessity of trapping hydrogen cyanide from biological samples, blood cyanide could be analyzed quickly by acidification only. Using this method, blood cyanide levels of fire victims were determined at autopsy. The serum values of cyanide ranged from 0.11 micrograms/ml to 18.12 micrograms/ml. However, a significantly higher cyanide content was detected in the left ventricular blood than in the right. This indicates that death was caused by the fire and suggests that the collecting point of the blood sample is an important factor in the determination of inhaled cyanide. There was a positive correlation between blood cyanide and carboxyhemoglobin contents.     

The role of hydrogen cyanide and carbon monoxide in fire casualties: a prospective study

Determinations of blood cyanide and carboxyhemoglobin concentrations were performed in 18 victims found dead in buildings after fires during a 2-year period. The results indicated that 50% of the victims had been exposed to toxic levels of hydrogen cyanide and 90% to toxic levels of carbon monoxide. Lethal concentrations of carbon monoxide were found in 83% of the victims. In one case a lethal blood cyanide but a non-toxic blood carboxyhemoglobin value was found. It is concluded that carbon monoxide appears to be more important than hydrogen cyanide as a toxic agent in the fire atmosphere, but cyanide poisoning without carbon monoxide poisoning may, under certain circumstances, be the cause of death in fire victims.     

Potential for error when assessing blood cyanide concentrations in fire victims.

The present study explores toxicologic significance of blood cyanide concentrations in fire victims. Headspace gas chromatography was used for cyanide detection. Analysis of blood samples from ten fire victims (postmortem interval = 8 h to 3 to 5 d) detected zero to 11.9 mg/L of cyanide and a large difference in cyanide concentrations among victims. Carboxyhemoglobin (COHb) saturation was in the range of 24.9 to 84.2%. To examine the effects of methemoglobinemia and postmortem interval on blood cyanide concentrations in fire victims, an experiment was carried out using rabbits as the animal model. The rabbits were sacrificed by intramuscular injection of 1 mL/kg 2% potassium cyanide 5 min after intravenous injection of 0.33 mL/kg of 3% sodium nitrite (Group A, n = 3) or physiological saline (Group B, n = 6). Average methemoglobin contents immediately before potassium cyanide administration were 6.9 and 0.8% in Groups A and B, respectively. Average cyanide concentrations in cardiac blood at the time of death were 47.4 and 3.56 mg/L, respectively. When blood-containing hearts of the rabbits (n = 3 for Group B) were left at 46 degrees C for the first 1 h, at 20 to 25 degrees C for the next 23 h and then at 4 degrees C for 48 h, approximately 85 and 46% of the original amounts of blood cyanide disappeared within 24 h in Groups A and B, respectively. After the 72-h storage period, 37 and 10%, respectively, of the original amounts of cyanide remained in the blood. When the other three hearts in Group B were left at 20 to 25 degrees C for the last 48 h without refrigeration, cyanide had disappeared almost completely by the end of the experiment. The present results and those published in the literature demonstrate that the toxic effects of cyanide on fire victims should not be evaluated based solely on the concentration in blood.     

Hydrogen cyanide and carbon monoxide in blood of convicted dead in a polyurethane combustion: a proposition for the data analysis

Carbon monoxide is a well-known toxic component in fire atmospheres. However, the importance of hydrogen cyanide as a toxic agent in fire causalities is under discussion. A tragic polyurethane mattress fire provoked death of 35 convicts in a prison (Unit I, Olmos, Penitenciary Service of Buenos Aires Province, Argentina), in 1990. There is no report of any investigation carried out with such a large amount of victims in Argentina. Carboxihemoglobin (COHb) and hydrogen cyanide (HCN) were quantified in victims blood to elucidate the cause of the death. Saturation of COHb ranged between 4 and 18%, and HCN 2.0-7.2mg/l. These latter values were higher than the lethal levels reported in literature. Other toxic components routinely measured (ethanol, methanol, aldehydes and other volatile compounds) gave negative results on the 35 cases. Neither drugs of abuse nor psychotropics were detected. Statistical chi(2) analysis was applied to find differences between HCN and COHb concentrations. Saturation of COHb and HCN in blood were not independent variables (chi(2)=8.25). Moreover, the ratio COHb/HCN was constant (0.47+/-0.04). In order to evaluate the contribution of each toxic to the diagnosis, a lethal index was defined for each toxic (LI(CO) and LI(HCN)). The most probable cause of death could be inferred by a suitable plot of both indexes. The results indicated that death in the 35 fire victims was probably caused by HCN, generated during the extensive polyurethane decomposition provoked by a rapid increase of temperature.     

Cyanide, carboxyhemoglobin and blood acid-base state in animals exposed to combustion products of various combinations of acrylic fiber and gauze

In order to examine the usefulness of blood cyanide concentrations as an indicator of whether or not a victim was alive at the start of a fire, blood cyanide concentrations were measured in the bodies that we autopsied in our institute between January 1986 and March, 1987. In the present study, bodies with advanced decomposition were excluded. Thirty-six bodies were included: cyanide as well as carboxyhemoglobin (COHb) were detected in four charred bodies found at the scene of a fire. On the other hand, cyanide was not detected in any of the remaining 30 bodies except in two cases suspected of having ingested a cyanide compound. Rats and rabbits were made to inhale the combustion products of various combinations of acrylic fiber (hydrogen cyanide generating material when heated) and gauze (carbon monoxide generating material when heated). The exposure to the combustion products was continued until death in the rat and until apnea in the rabbit. The concentration of hydrogen cyanide in the exposure chamber and that of blood cyanide, at the time the animal died, correlated with the amount of acrylic fiber heated. In addition to differences in blood COHb and cyanide concentrations, there were also differences in blood gas concentrations between the acrylic fiber and the gauze groups. When the rabbits were switched to room air after the occurrence of apnea, the blood gas value began to normalize.     

The effect of sodium fluoride on the stability of cyanide in postmortem blood samples from fire victims

Assigning a level of significance to cyanide concentrations found in the blood of fire victims is often hampered by the fact that cyanide is inherently unstable in cadavers and in stored blood samples. A few researchers have proposed that sodium fluoride can be used to minimize the instability of cyanide in blood samples; however, controlled studies have not been performed to support validation of this hypothesis. To test the sodium fluoride hypothesis, both treated and control blood samples from 14 autopsied fire victims were tested over a 25-30 day period. A 2% concentration of sodium fluoride was added to the blood samples at the start of testing and the samples were refrigerated between testing intervals. Cyanide concentrations in the treated and control samples were measured between 9 and 11 days post treatment and between 25 and 30 days post treatment. A statistically significant difference was not present between blood cyanide concentrations in treated and control samples between 9 and 11 days. During this time period, although there were small statistically significant increases in both treated and untreated samples the fluctuations were minor. Since the treated and control samples did not exhibit instability between 9 and 11 days, it is not surprising that the sodium fluoride appeared to have no effect. However, a statistically significant difference between blood cyanide concentrations in treated and control samples was observed between 25 and 30 days. Those samples treated with sodium fluoride showed a reduction in blood cyanide variability with virtually no overall change, over a 25-30 day period when compared to control samples, while unconditioned samples showed a significant, average increase of 35%. Based on the findings of this study, it is recommended that 2% sodium fluoride be added to blood samples obtained from fire victims to reduce cyanide instability due to bacteriological activity.     

The toxicological examination of the victims of the British Air Tours Boeing 737 accident at Manchester in 1985

The results of toxicological analyses of the body fluids of the victims from the accident involving the British Air Tours Boeing 737 in August 1985 are presented for carboxyhemoglobin, cyanide, and volatiles. All the victims except one had raised concentrations of carbon monoxide. All the victims had raised concentrations of cyanide. All the victims showed the presence of volatile substances in the blood. Autopsies revealed that all the victims had carbon particles in the trachea and bronchi. Thus, all the victims must have inhaled fire products in the burning aircraft cabin. Six victims had concentrations of carbon monoxide or cyanide in the blood that were neither fatal nor incapacitating; therefore, it is reasonable to suggest that these six victims survived for a comparatively short time and that there may have been other causes, in addition to toxic fumes, for their deaths. The other 48 victims must have survived long enough in the fire to accumulate incapacitating or fatal concentrations of carbon monoxide or cyanide or both. The effects of these substances found in the blood of each of the 48 victims must have combined to produce an insurmountable impediment to escape from the aircraft.     

Cyanide quantification in post-mortem biological matrices by headspace GC-MS

Cyanide is a powerful chemical asphyxiant found in some forensic cases following voluntary (suicide) or involuntary ingestion (fire, accidental exposure). A quantification method for cyanide that is specifically suited to post-mortem forensic purposes was developed. Determination was performed by headspace gas chromatography coupled to mass spectrometry using a GS-GASPRO column on an HP-6890 gas chromatograph with an HP-5973N mass detector. The biological sample was treated with an internal standard, frozen, glacial acetic acid was added and the sample was then incubated at 60°C for 15min. The headspace was sampled with a disposable syringe, and analyzed to quantify hydrogen cyanide. Isotopically labeled cyanide ((13)C(15)N) was used as the internal standard to minimize matrix effect and sampling error. The method produced an extended linear dynamic range (0.07-50μg/mL), and a method detection limit of 0.02μg/mL. Identical calibration curves were obtained when blood, gastric contents and aqueous solutions were used as the calibration standard matrix. This method was also successful in quantitating cyanide in gastric contents, one of the most variable biological fluids. The method has been validated and is being used for current forensic cases such as fire victims and suicides.     

"First pass phenomenon" of inhaled gas in the fire victims

We investigated the differences in the levels of carboxyhemoglobin (COHb), cyanide (HCN) and petroleum fuels (gasoline and kerosene) between left and right ventricular bloods from fire victims. COHb was slightly, and HCN and petroleum fuels were markedly higher levels in the left than those in the right. These effects were so called 'first pass phenomena' due to the circulation, diffusion and metabolization before the deaths of fire victims.     

High cyanide level in a homicide victim burned after death: evidence of post-mortem diffusion.

Elevated levels of carbon monoxide and cyanide serve as evidence of intravital burning in fire victims. Hydrogen cyanide is released by combustion of nitrogen-containing organic material such as plastics and wool. We present a case of a man who died of haemopneumothorax caused by a stab wound. According to several eye witnesses the body was wrapped in a plastic sheet and burned 2 days after death with the aid of gasoline. No coal pigment was observed in the mucosa of the upper airways at autopsy. The blood sample taken from the pulmonary vessels 6 days after death disclosed a level of blood carboxyhaemoglobin of 4% and of blood cyanide of 10 mg/l. The low carboxy-haemoglobin level was consistent with the smoking habits of the victim. The thoracic cavity had been opened by burning of the intercostal soft tissue. This allowed hydrogen cyanide gas to enter the thoracic cavity and diffuse into the blood probably causing the high blood-cyanide level.     

Analysis of carboxyhemoglobin and cyanide in blood from victims of the Dupont Plaza Hotel fire in Puerto Rico

Ninety-seven people died from a fire that occurred in the Dupont Plaza Hotel in Puerto Rico on 31 Dec. 1986. All, except four who died later in the hospital, were found dead at the scene. All of the fatalities at the hotel (except for eight) were burned beyond recognition. Blood from seventy-eight of the victims was screened for carboxyhemoglobin at the Institute for Forensic Sciences in Puerto Rico and was then sent to the National Institute of Standards and Technology, Gaithersburg, Maryland, for analysis of carboxyhemoglobin and cyanide concentrations. The blood data indicated that carbon monoxide and hydrogen cyanide, singly or combined, were probably not responsible for the majority of the deaths that occurred in the badly burned victims. On the other hand, the significantly higher carboxyhemoglobin in the nonburned victims indicated that carbon monoxide alone or combined with hydrogen cyanide probably played a major role in the cause of their deaths.     

Significance of hydrocyanic acid formation during fires]

Cyanide concentrations of blood samples from fire victims autopsied in the Institute of Legal Medicine, Munich, have been determined. In 25% of 48 analyzed cases cyanide concentrations from 0.52 microgram to 6.24 microgram Cyanide/ml blood have been detected. These results are compared to former studies and the higher mean level in our collective is emphasized. The importance of hydrocyanid acid in the toxicity of fire gases is evidently greater, than assumed. Hydrocyanic acid may be produced from nitrogen continaing polymers during combustion. The quote of these polymers in clothing, furniture, and also in equipment of cars is increasing. Therefore, it is necessary to take more notice of the formation of hydrocyanic acid during combustion, even though carbon monoxide is in general the main toxic agent in fire gases.     

Application of computational fluid dynamics modelling in the process of forensic fire investigation: problems and solutions

Fire modelling has been gaining more and more interest into the community of forensic fire investigation. Despite an attractiveness that is partially justified, the application of fire models in that field of investigation rises some difficulties. Therefore, the understanding of the basic principles of the two main categories of fire models, the knowledge of their effective potential and their limitations are crucial for a valid and reliable application in forensic science. The present article gives an overview of the principle and basics that characterise the two kinds of fire models: zone models and field models. Whereas the first ones are developed on the basis of mathematical relation from empirical observations, such as stratification of fluid zones, and give a relatively broad view of mass and energy exchanges in an enclosure, the latter are based on fundamentals of fluid mechanics and represent the application of Computational Fluid Dynamics (CFD) to fire scenarii. Consequently, the data that are obtained from these two categories of fire models differ in nature, quality and quantity. First used in a fire safety perspective, fire models are not easily applied to assess parts of forensic fire investigation. A suggestion is proposed for the role of fire modelling in this domain of competence: a new tool for the evaluation of alternative hypotheses of origin and cause by considering the dynamic development of the fire. An example of a real case where such an approach was followed is explained and the evaluation of the obtained results comparing to traces revealed during the on-site investigation is enlightened.     

Expert significance of evaluating volatile non-metal combustion products in analysis of blood from subjects dead in fire

Qualitative and quantitative composition and time course of formation of toxic substances during burning of polymers have been studied for two polymers, P-53-P and STBO. The most toxic gaseous products of nitrogen combustion are listed: hydrogen cyanide, nitrites, acid nitriles, aldehydes, aromatic carbohydrates, alkanes, and carbon oxide. Two expert conclusions are presented, including analysis of blood and viscera from subjects who died during fire. The following toxic substances were detected: carbon oxide, hydrogen cyanide, nitrites, acrylonitrile, acetonitrile, benzene, and methane in one case and the same products except carbon oxide in the other. The author considers that the list of tested toxic substances should be extended, particularly in cases with negative results of blood testing for carbon oxide and carboxyhemoglobin.     

The ‘quarter-power equation’ for resistive heating faults

The ubiquitous nature of electricity makes an electrical fault a primary consideration at the start of almost every fire investigation. Principal amongst the types of faults that can cause a fire is the resistive heating fault. While methods for calculating the maximum power transfer from a supply to a load are well known to electrical engineers, especially in the fields of radio and audio engineering, their useful application for fire investigation has not been explored publicly. A power transfer relationship is re-worked here by considering the original circuit as the supply and the fault as the load. This results in the ‘quarter-power equation’ for application in fire investigation, which can help to assess whether this particular means of ignition, the resistive heating fault, is viable for the case being investigated. This article provides analysis and guidance on the derivation, use and limitations of the ‘quarter-power equation’ and shows how it was useful in eliminating a possible cause of fire under consideration in a particular investigation.     

Analysis of hydrogen cyanide in air in a case of attempted cyanide poisoning

A 32-year-old man attempted to poison his ex-girlfriend with hydrogen cyanide by hiding the pesticide Uragan D2 in her car. During the police investigation, chemical analysis of the air inside the car was performed. Hydrogen cyanide was detected through on-site air analysis using a portable Fourier transform infrared (FTIR) spectroscopy gas analyzer and colorimetric gas detection tubes. Furthermore, impinger air-sampling was performed for off-site sample preparation and analysis by gas chromatography-mass spectrometry (GC-MS). All three independent techniques demonstrated the presence of hydrogen cyanide, at concentrations of 14-20ppm. Owing to the high volatility of hydrogen cyanide, the temperature and the time since exposure have a substantial effect on the likelihood of detecting hydrogen cyanide at a crime scene. The prevailing conditions (closed space, low temperature) must have supported the preservation of HCN in the car thus enabling the identification even though the analysis was performed several days after the hydrogen cyanide source was removed. This paper demonstrates the applicability of combining on-site FTIR measurements and off-site GC-MS analysis of a crime scene in order to ensure fast detection as well as unambiguous identification for forensic purposes of hydrogen cyanide in air.     

Cyanide and amygdalin as indicators of the presence of bitter almonds in imported raw almonds

Abstract: Consumer complaints received by the U.S. Food and Drug Administration in August 2010 about raw organic almonds tasting “bitter” opened an investigation into the presence of bitter almonds in the imported product. Bitter almonds (Prunus amygdalus) contain the cyanogenic glucoside amygdalin, which hydrolyzes to produce cyanide. Ultraviolet–visible spectrophotometry was used to detect and quantitate cyanide, and liquid chromatography‐mass spectrometry was utilized to detect amygdalin in the submitted samples. Control bitter almonds were found to contain 1.4 mg cyanide/g and an estimated level of 20–25 mg amygdalin/g. The questioned samples contained between 14 and 42 μg cyanide/g and were positive for the presence of amygdalin. Sweet almonds were found to be negative for both compounds, at levels of detection of 4 μg cyanide/g and 200 μg amygdalin/g.     

A study on house fire victims: age, carboxyhemoglobin, hydrogen cyanide and hemolysis.

Correlation among age, concentrations of carboxyhemoglobin and hydrogen cyanide, oxygen density and hemolysis were studied in 120 house fire victims. Victims aged over 60 years comprised approximately 50% of the pooled subjects. Blood samples were mainly collected from the left ventricle, but sometimes from both the right and left ventricles. The concentration of carboxyhemoglobin ranged from 1-95%, of which 71 persons (59.7%) died with carboxyhemoglobin concentrations below 60%. Carboxyhemoglobin concentrations below 10% were found in 9 persons (7.5%). Most of these cases involved the elderly persons. In this paper, we report on the death of elderly victims as a result of low carboxyhemoglobin concentrations. A significant correlation of blood carboxyhemoglobin concentrations existed between the right and left ventricles. The concentration of carboxyhemoglobin in the left ventricle was significantly higher than that in the right. Two out of 31 victims whose hydrogen cyanide concentrations were determined, succumbed to hydrogen cyanide poisoning, having a high concentration of hydrogen cyanide and a low concentration of carboxyhemoglobin. On analysis, oxygen density was found to be low in 13 persons. A negative correlation was shown between carboxyhemoglobin concentration and hemolysis. Inasmuch as hemolysis may indicate the extent of heat dissociation, hemolysis should provide an index of carbon monoxide dissociation from carboxyhemoglobin. In the present study of victims, possible causes of death i.e., carbon monoxide gas poisoning, hydrogen cyanide poisoning, oxygen deprivation, burning, shock due to burns and others were estimated. The survival time for elderly victims was considered to be short.     

Inhalation heat shock]

Using the example of a recent case, we examine the possibilities of trace analysis and fire protection techniques in the investigation of fire deaths. The effective cooperation between forensic pathologists and fire inspectors is presented and we discuss the mathematical-physical uses of fire simulation calculations. We review the trace analysis results and discuss the variety of technical possibilities of modern engineering techniques for fire protection.     

Accidental fire deaths. The 5-year Metropolitan Dade Country experience from 1979 until 1983

Accidental fire deaths that occurred in Metropolitan Dade County, encompassing Miami, Fla., USA, during the years from 1979 until 1983 were studied. A total of 108 cases were collected, representing 6.1% of the nontraffic-related accidents during this 5-year period, and analyzed as to the age, race, sex, cause of death of the victim along with the blood alcohol content at autopsy, toxicologic analysis, location of the fire, and how the fire started. Essentially, the victims ages cluster in the 0-5 years and does 70 years, although other age groups are evenly distributed. A white male population predominates. The cause of death is listed as either smoke inhalation or thermal injury. Most toxicologic analyses were not performed due to the high rate of fire rescue intervention and at least emergency room hospitalization prior to death. Carboxyhemoglobin levels, when analyzed, ranged from less than 20% to over 80%. In a smaller group of cases, cyanide was analyzed for and was found negative (or none detected) in the majority of the cases. More fires occurred in residences and started due to cigarette smoking or electric malfunction.