Determination of Cyanide by Microdiffusion Technique Coupled to Spectrophotometry and GC/NPD and Propofol by Fast GC/MS-TOF in a Case of Poisoning.

A man was found dead in a hotel located near Rome (Italy). The man was still holding a syringe attached to a butterfly needle inserted in his left forearm vein. The syringe contained a cloudy pinkish fluid. In the hotel room the Police found a broken propofol glass vial plus four sealed ones, an opened NaCl plastic vial and six more still sealed, and a number of packed smaller disposable syringes and needles. An opened plastic bottle containing a white crystalline powder labeled as potassium cyanide was also found. Systematic toxicological analysis (STA), carried out on blood, urine and bile, evidenced only the presence of propofol in blood and bile. So the validated L-L extraction protocol and the GC/MS-TOF method for the confirmation of propofol in the biological fluids optimized in our laboratory was applied to blood, urine and bile. The concentration of propofol resulted to be 0.432 µg/mL in blood and 0.786 µg/mL in bile. The quantitative determination of cyanide in blood was carried out by microdiffusion technique coupled to spectrophotometric detection obtaining a cyanide concentration of 5.3 µg/mL. The quantitative determination was then confirmed by GC/NPD and the concentration of cyanide resulted to be 5.5 µg/mL in blood and 1.7 µg/mL in bile. Data emerging from autopsy findings, histopathological exams and the concentrations of cyanide suggested that death might be due to poisoning caused by cyanide, however, respiratory depression caused by propofol could not be excluded.

Evaluation of the Removal of Potassium Cyanide and its Toxicity in Green Algae (Chlorella vulgaris).

To evaluate the removal of potassium cyanide (KCN) and its toxicity in algae, an initial comprehensive analysis was performed with Chlorella vulgaris. The algae showed potential removal capability for KCN, with the maximal removal rate of 61%. Moreover, effects of KCN on growth, cellular morphology and antioxidant defense system of C. vulgaris were evaluated. Cell number and chlorophyll a content decreased in most cases, with the maximal inhibition rates of 48% and 99%, respectively. The 100 mg L- 1 KCN seriously damaged the algal cell membrane. Additionally, activity of superoxide dismutase (SOD) was promoted by KCN exposure among 0.1-50 mg L- 1 and inhibited by 100 mg L- 1 KCN, while the malondialdehyde (MDA) content gradually decreased in C. vulgaris with increasing exposure concentration compared to the control. The present study reveals that C. vulgaris is useful in bio-treatment of cyanide-contaminated aquatic ecosystem, except in high concentrations which would cause overwhelming effects.

Detecting total toxicity in water using a mediated biosensor system with flow injection.

A novel total toxicity detection method based on a mediated biosensor system with flow injection (MB-FI) was developed to rapidly and reliably detect respiration inhibitors (i.e., As2O3, KCN, salicylic acid (SA), 2,4-dintirophenol (DNP)) in water. The mediated biosensor toxicity assessment using microorganisms immobilized in calcium alginate filaments can greatly simplify the testing process and save time. In the MB-FI system, ferricyanide together with a respiration inhibitor was injected into the bioreactor, inhibiting the respiration of the immobilized microorganisms. The degree of inhibition was measured by determining the ferrocyanide generated in the effluent, expressed as the 50% inhibition concentration (IC50). The IC50 values for the four respiration inhibitors obtained using this method were comparable to those obtained using the classic method, confirming that this approach is an alternative alert method. More importantly, this constructed biosensor system with flow injection will facilitate the application and commercialization of this toxicity monitoring technology.

An intramolecular crossed-benzoin reaction based KCN fluorescent probe in aqueous and biological environments.

A turn-on fluorescent probe was designed for selective cyanide anion sensing in aqueous and biological environments. The probe underwent an intramolecular crossed-benzoin reaction in the presence of KCN to expel the fluorophore resorufin. This probe was sensitive to KCN concentrations as low as 4 nM in aqueous media.

Amperometric inhibition biosensors based on horseradish peroxidase and gold sononanoparticles immobilized onto different electrodes for cyanide measurements.

New biosensors based on inhibition for the detection of cyanide and the comparison of the analytical performances of nine enzyme biosensor designs by using three different electrodes: Sonogel-Carbon, glassy carbon and gold electrodes were discussed. Three different horseradish peroxidase immobilization procedures with and without gold sononanoparticles were studied. The amperometric measurements were performed at an applied potential of -0.15V vs. Ag/AgCl in 50mM sodium acetate buffer solution pH=5.0. The apparent kinetic parameters (Kmapp, Vmaxapp) of immobilized HRP were calculated in the absence of inhibitor (cyanide) by using caffeic acid, hydroquinone, and catechol as substrates. The presence of gold sononanoparticles enhanced the electron transfer reaction and improved the analytical performance of the biosensors. The HRP kinetic interactions reveal non-competitive binding of cyanide with an apparent inhibition constant (Ki) of 2.7µM and I50 of 1.3µM. The determination of cyanide can be achieved in a dynamic range of 0.1-58.6µM with a detection limit of 0.03µM which is lower than those reported by previous studies. Hence this biosensing methodology can be used as a new promising approach for detecting cyanide.

A stable in vitro method for assessing the toxicity of potassium cyanide and its antidote.

BACKGROUND: Hydrogen cyanide possesses a high acid-dissociation constant of 9.14, favoring its vaporization and depletion from the culture media at physiological pH, which may cause the cyanide toxicity unstable in vitro. OBJECTIVE: We investigated whether adjustment of culture medium pH stabilizes cyanide concentration and decreases the effective concentration of potassium cyanide (KCN). METHODS: Murine fibroblast cells were exposed to different concentrations of KCN in media maintained at pH 7.4 or 9.2, in the presence or absence of hydroxocobalamin. After incubation for 1 h, we evaluated medium pH, cyanide concentration, cytochrome activity, and cell viability. RESULTS: Cyanide concentration decreased to 18.8% in pH 7.4 medium compared to 83.2% in pH 9.2 medium. A significant decrease in cytochrome activity was observed at 40 mM and 1.25 mM KCN in pH 7.4 and pH 9.2 media, respectively. In pH 9.2 medium, dose-dependent cytotoxicity of KCN and antidotal effects of hydroxocobalamin were observed. CONCLUSION: Adjustment of culture medium pH to 9.2 could stabilize cyanide concentration and decrease the effective concentration of KCN, allowing stable evaluation of KCN toxicity and antidotal efficacy.

A new biological test utilising the yeast Saccharomyces cerevisiae for the rapid detection of toxic substances in water.

This study evaluates the toxic effects of five substances (atropine, fenitrothion, potassium cyanide, mercuric chloride and lead nitrate) on the yeast Saccharomyces cerevisiae. It describes a new biological toxicity test based on inhibition of S. cerevisiae viability and compares it with two standard toxicity tests based on Daphnia magna mobility inhibition (EN ISO 6341) and Vibrio fischeri bioluminiscence inhibition (EN ISO 11348-2). The new biological test -S. cerevisiae lethal test - is cheaper and 24 times faster than the D. magna test. The test speed is comparable with the V. fischeri test but the new test is more sensitive for some substances. The test indicates reliably the presence of all used toxicants in water in concentrations which are significantly lower than the concentration in toxic or lethal doses for man. Therefore, this new toxicity test could be proposed for rapid detection of toxic substances in water.

Simultaneous screening of glutathione and cyanide adducts using precursor ion and neutral loss scans-dependent product ion spectral acquisition and data mining tools.

Drugs can be metabolically activated to soft and hard electrophiles, which are readily trapped by glutathione (GSH) and cyanide (CN), respectively. These adducts are often detected and structurally characterized using separate tandem mass spectrometry methods. We describe a new method for simultaneous screening of GSH and CN adducts using precursor ion (PI) and neutral loss (NL) scans-dependent product ion spectral acquisition and data mining tools on an triple quadrupole linear ion trap mass spectrometry. GSH, potassium cyanide, and their stable isotope labeled analogues were incubated with liver microsomes and a test compound. Negative PI scan of m/z 272 for detection of GSH adducts and positive NL scans of 27 and 29 Da for detection of CN adducts were conducted as survey scans to trigger acquisition of enhanced resolution (ER) spectrum and subsequent enhanced product ion (EPI) spectrum. Post-acquisition data mining of EPI data set using NL filters of 129 and 27 Da was then performed to reveal the GSH adducts and CN adducts, respectively. Isotope patterns and EPI spectra of the detected adducts were utilized for identification of their molecular weights and structures. The effectiveness of this method was evaluated by analyzing reactive metabolites of nefazodone formed from rat liver microsomes. In addition to known GSH- and CN-trapped reactive metabolites, several new CN adducts of nefazodone were identified. The results suggested that current approach is highly effective in the analysis of both soft and hard reactive metabolites and can be used as a high-throughput method in drug discovery.

KCN sensor: unique chromogenic and 'turn-on' fluorescent chemodosimeter: rapid response and high selectivity.

An indole conjugated coumarin 1 for KCN chemodosimeter has been prepared and displayed considerable dual changes in both absorption (blue-shift) and emission (turn-on) bands exclusively for KCN. DFT/TDDFT calculations support that the fluorescence enhancement of 1-KCN is mainly due to blocking of the ICT process.

Highly cooperative ion-pair recognition of potassium cyanide using a heteroditopic ferrocene-based crown ether-trifluoroacetylcarboxanilide receptor.

A heteroditopic receptor having crown ether and trifluoroacetylcarboxanilide groups selectively recognizes both potassium and cyanide ions in acetonitrile with an association constant of as high as Ka = 1.9 x 10(7) M(-1) through a highly cooperative ion-pair interaction, resulting in two orders of magnitude enhancement in the binding affinity.

Glass distilling collector applied for HCN recovery from submerged culture broth and fruiting body of Pleurotus eryngii for identification and quantification.

Detection and surveillance of food commodities containing cyanide is a crucial issue of food safety. In this study, five strains of Pleurotus eryngii (P. eryngii) were grown in submerged culture of yeast malt broth (YMB) with the suspected production of HCN. A safety-warranted U-bent glass distilling collector with three enlarged bulbs on each arm was designed to recover the broth vapor. When AgNO(3) solution was used as an absorbent to interact with the vapor, a white precipitate was formed. The precipitate was isolated and identified as AgCN by FT-Raman spectroscopic analysis. When the absorbent was substituted by KOH, after evaporation to dryness, dissolved in D(2)O, and followed by (13)C-NMR analysis, a KCN spectrum was achieved. Formation of AgCN and KCN confirmed HCN production in the broth by P. eryngii. When a sodium picrate solution (1.4%) was used as an absorbent and various authentic KCN solutions were applied for distillation and followed by absorbance determination at 510 nm, a linear dose-dependent relationship was obtained and the procedure was applied for HCN quantification of the marketed P. eryngii mushrooms (fruiting body). As estimated, 67.3% of the products contained HCN less than 1.0 mg/kg, 17.3% between 1.0 and 2.0 mg/kg, and 15.4% higher than 2.0 mg/kg. When the mushrooms were sliced and cooked in water at 95 degrees C for 6 min, 89.1% of the original HCN was lost. When the P. eryngii strains were respectively grown by submerged cultivation in YMB or YMB supplemented with 2.5% glycine for 16 days, HCN content was slightly higher in the latter than in the former for each strain.

Detoxification of cyanide by woody plants.

Vascular plants possess an enzyme system that detoxifies cyanide by converting it to the amino acid asparagine. This paper examines the potential of three woody plants from the Salicaceae family to degrade cyanide. Pre-rooted trees were grown in carefully designed bioreactors with aqueous solution spiked with potassium cyanide at 23.0 +/- 1 degree C for a maximum of 144 h. Cyanide concentrations ranged from 0.95 to 1.15 CN mg/L. Cyanide in water and in plant tissues was analyzed spectrophotometrically. Results from the investigation indicated that significant reduction of aqueous cyanide was found during the presence of plants in all treatments. Little amounts of applied cyanide were detected in the tissues of plants, mainly in roots and bottom stem. Cyanide remaining in tissues varied with the species of plants, despite similar periods of exposure. The data also indicated that photolysis, hydrolysis, and microbial degradation were not occurring and that volatilization was minimal. In conclusion, transport and metabolism of cyanide in plants is most likely.

Survival of fish upon removal of cyanide from water.

The effects of potassium cyanide and the removal of cyanide from water in vivo on the survival of fish were investigated. This research was initiated because of the catastrophe that took place at the end of January 2000 in the Carpathian basin, when an enormous amount of cyanide pollution swept through the Samos and Tisza rivers, and then to the Danube. Since nothing was done against the disaster, we have suggested a chemical solution to remove cyanide from waterways (Chem. Innovat. 30 (2000b) 53). Based on experiments, we describe that the most effective and harmless way to remove cyanide and to save the lives of fish from 40 to 160 x the lethal doses of cyanide is to use carbogen gas containing 5% carbon dioxide and 95% oxygen followed by aeration with air.

Application of toxicity identification evaluation procedures to an effluent from a nitrogenous fertilizer plant in China.

The integrated method combining chemistry and toxicology, toxicity identification evaluation (TIE), was conducted to identify key toxicants in an effluent from a nitrogen fertilizer plant in China. Toxicity characterization, phase I of TIE, revealed that the suspected toxicant in the effluent was an anion that could be changed into a volatile acid. The results of toxicity identification and confirmation procedures indicated potassium cyanide to be the primary toxicant in the effluent.

Elution and decomposition of cyanide in soil contaminated with various cyanocompounds.

Standard soil samples contaminated with various standard cyanocompounds were prepared. Column elution experiments and analyses were conducted. Compounds with an easy capacity for dissociation to ions, such as KCN and potassium hexacyanoferrate(III), were found to be eluted by forming free cyanide even in fresh water. Hexacyanoferrate(II) salts, such as potassium hexacyanoferrate(II) and iron(III) hexacyanoferrate(II), were found not to be dissociated in water, but were dissociated and diffused under alkaline conditions (pH >13). Hexacyanoferrate(II) ion was found to be more easily dissociated in water with a higher pH. Column tests as above were also conducted for soil samples taken from a former paint ink factory using iron(III) hexacyanoferrate(II), cyanogen chloride, potassium cyanate, copper cyanide, as well as potassium cyanide, as raw materials. It was demonstrate that iron(III) hexacyanoferrate(II) was dissociated and eluted under alkaline conditions. The elution rate was reduced when the contaminated soil was sandwiched with standard soil layers.Further, it was found that the Fe(CN)(6)(4-) ion eluted with NaOH from hexacyanoferrate acid in soil, were easily decomposed into cyanic acid or other byproducts by UV with the addition of ozone and H(2)O(2).

Cyanide-reactive sites in cytochrome bd complex from E. coli.

Cyanide reacts with cytochrome bd from E. coli in an 'aerobically oxidized' state (mainly, an oxygenated complex b558(3+) b595(3+) d(2+)-O2), bringing about (i) decomposition of the heme d2+ oxycomplex (decay of the 648 nm absorption band) and (ii) extensive red shift in the Soret region accompanied by minor changes in the visible range assigned to ferric heme b595. MCD spectra show that the Soret red shift is associated with heme b595(3+) high-to low-spin transition. This is the first unambiguous demonstration that heme b595 can bind exogenous ligands. No reaction of cyanide with b558 is observed. In about 70% of the enzyme which forms the cyano complex, the spin-state transition of b595 decay of heme d oxycomplex match each other kinetically (keff ca. 0.002 s-1 at 50 mM KCN, pH 8.1, 25 degrees C). This points to an interaction between the two hemes. The concerted binding of cyanide to d3+ and b595(3+), perhaps as a bridging ligand, is probably rate-limited by d2+ oxycomplex autoxidation. In the remaining 30% of the isolated bd, there is a rapid phase of cyanide-induced b595 spin-state transition which can be tentatively assigned to that proportion of the enzyme in which heme d is initially in the ferric rather than ferrous-oxy form.

[A photometric method of analysis of cyanides]. / Fotometricheskii sposob opredeleniia tsianidov.

The contributors propose a selective photometric technique for detection of sodium and potassium cyanides in different washings, sewage, overalls extraction. The technique was based on the formation of a polymetyne dye, and barbituric acid and pyridine reactions. It proved efficient in detecting the cyanides within 0.01-0.054 mg/dm3, 1-100 mg/l, 0.08-1.24 mg/dm3 with deviations not exceeding 23%. Duration of the test was limited to 25-30 min.

Effects of exposure to single or multiple combinations of the predominant toxic gases and low oxygen atmospheres produced in fires.

The toxicity of single and multiple fire gases is studied to determine whether the toxic effects of the combustion products from materials can be explained by the toxicological interactions (as indicated by lethality) of the primary fire gases or if minor, more obscure gases need to be considered. LC50 values for Fischer-344 rats have been calculated for the individual gases, carbon monoxide (CO), hydrogen cyanide (HCN), or decreased oxygen (O2), for 30-min exposures plus relevant postexposure periods using the NBS Toxicity Test Method. Combination experiments with CO and HCN indicate that they act in an additive manner. Synergistic effects have been found when the animals are exposed to certain combinations of CO and carbon dioxide (CO2). Five percent CO2 raised the threshold for deaths due to hypoxia and decreased the LC50 of HCN. Decreasing the O2 concentration in the presence of various mixtures of the other major fire gases increased the toxicity even further. A comparison of the concentrations of the major combustion products generated from a number of polymeric materials at their LC50 (30-min exposure plus 14-day postexposure) values with the combined pure gas results indicates that, in most cases, the observed toxicity may be explained by the toxicological interactions of the examined primary toxic fire gases. These results provide necessary information for the computer model currently being developed at the Center for Fire Research to predict the toxic hazard that people will experience under various fire scenarios.

Synthesis and quality assurance of [11C]alpha-aminoisobutyric acid (AIB), a potential radiotracer for imaging and amino acid transport studies in normal and malignant tissues.

Carbon-11 labeled alpha-aminoisobutyric acid (AIB), a synthetic amino acid, was prepared by the modified Bucherer-Strecker amino acid synthesis from acetone, ammonium carbonate and [11C]KCN in the presence of carrier KCN. This method results in the labeling of AIB in the carboxyl group. The label is stable in this position because AIB is not a metabolized after cellular uptake. AIB is rapidly accumulated in viable cells including malignant cells. Since it is a non-metabolized amino acid, AIB offers the possibility of studying amino acid transport in vivo without interference by radiolabeled metabolic products. Radiochemical yields of [11C]AIB of 35-60% have been obtained in 70-80 min with radiopurities greater than 99%. Carrier added syntheses gave 15-25 mCi of [11C]AIB with specific activities of 0.3 Ci/mmol. Our quality control program which insures that [11C]AIB is suitable for imaging studies in patients with cancer includes HPLC analyses of product identity and purity, apyrogenecity and isotonicity assays, and a sensitive test for cyanide.