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Rate of Metabolism of Cyanide to Thiocyanate in Saliva After Smoking. By: Matt Herring Deanne Seymour and Bettylou Wahl. What is Cyanide?. Common forms: HCN, NaCN, KCN Found in foods such as cassava, lima beans, almonds, and apples Produced by certain bacteria and fungi

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rate of metabolism of cyanide to thiocyanate in saliva after smoking

Rate of Metabolism of Cyanide to Thiocyanate in Saliva After Smoking


Matt Herring

Deanne Seymour


Bettylou Wahl

what is cyanide
What is Cyanide?
  • Common forms: HCN, NaCN, KCN
  • Found in foods such as cassava, lima beans, almonds, and apples
  • Produced by certain bacteria and fungi
  • Enters the body through ingestion, inhalation, and absorption
hydrogen cyanide
Hydrogen Cyanide
  • Colorless gas
  • Almond scent
  • BP: 25.6 C
  • Enters the body through inhalation
  • Toxic gas present in cigarette smoke
  • Released in metallurgy, electroplating, metal cleaning processes and car exhaust
  • Used for fumigation of dry foods such as cereals, seeds, nuts, and tobacco
  • Used for disinfestation of buildings
effects of hcn on the body
Effects of HCN on the body
  • Chronic low exposure causes neurological, respiratory, cardiovascular, and thyroid effects
    • breathing difficulties, heart pains, vomiting, blood changes, headaches
  • Long term exposure causes central nervous system effects
    • weakness of digits, difficulty walking, dimness of vision, deafness
  • High levels of exposure in a short amount of time harms the brain and heart and may cause coma and death
cigarette smoke
Cigarette Smoke
  • Cigarettes are a large source of cyanide
  • Cyanide is not present in actual cigarettes, but is formed through combustion and found in the smoke
  • Cyanide levels in inhaled cigarette smoke range from 10 to 400 micrograms per cigarette
  • Cyanide is metabolized to less toxic thiocyanate through sulfuration with thiosulfate by mitochondrial rhodanase in the liver
  • CN- + S2O3-2 SCN- + SO3-2
  • Thiocyanate is present normally in human saliva at approximately [0.01%]
  • Thiocyanate levels in saliva have been found correlate with cyanide intake
methods for determining cyanide and thiocyanate levels
Methods for determining cyanide and thiocyanate levels
  • HS-GC (head-space gas chromatography)
  • Spectrophotometric Konig method
    • Thiocyanate ion (SCN-) reacts with iron Fe3+ to yield FeSCN2+ complex which can be detected spectrophotometrically (at 448 nm)
    • The complex exhibits a red/orange color that becomes darker with greater concentration
    • UV-VIS (HP) Instrument is used to measure the absorption of FeSCN2+ complex, which correlates to the [SCN-]
previous research and studies
Previous research and studies
  • Saliva thiocyanate levels of smokers, non-smokers, and second hand smokers have been studied
  • Smokers have been found to have higher overall levels of thiocyanate than non-smokers (Lahti et. al. 1999)
  • Establishing an accurate calibration curve
  • Monitoring peoples’ diets for testing
  • Storing the samples
  • Obtaining a large enough sample size
  • Finding a strong control when there are many variables
our research
Our Research
  • Initially, saliva samples from smokers and non-smokers were analyzed (Juarez 2004)
  • In order to confirm past research, we set out to see if there were any significant differences in thiocyanate levels between smokers and non-smokers
method for preparing saliva
Method for preparing saliva
  • Obtain 2.5 mL of saliva
  • Centrifuge at 12,000 rpm for 12 min
  • Remove and centrifuge clear liquid again at 12,000 rpm for 12 min
  • Add 0.5 mL of centrifuged saliva to 9.5 mL of 0.0019 M Fe(NO3)3
  • Measure absorption at 448 nm in spectrophotometer
instrumental detection levels
Instrumental Detection Levels

HP 8452A Diode Array Spectrophotometer

establishing standard curve
Establishing Standard Curve
  • Beer’s Law: A=abc
  • Used to determine the concentration from the experimental absorption level values
  • Established using five known concentrations of FeSCN as standards
  • Curve checked for accuracy

SCN- concentration (M)

  • Results did not show higher overall thiocyanate levels for smokers
  • Not enough samples analyzed
  • Problem with our methods?
  • Initially samples were take right after the subject smokes
  • Did this allow ample time for cyanide to be metabolized after smoking?
new scope of investigation
New Scope of Investigation
  • Set to find any change in thiocyanate levels over time after the subject smokes
  • If any changes are observed, then the variable of time must be taken into account
round 2 rate of metabolism
Round 2: Rate of Metabolism
  • Saliva samples were taken before smoking and after smoking at set time intervals (initially, 30 minutes, 60 minutes)
  • Changes in thiocyanate concentrations over time will allow us observe both the rate of metabolism and degradation of thiocyanate in the saliva
future goals
Future Goals
  • Reconfirm the rate study of metabolism of cyanide to thiocyanate
  • Once again take up our previous research involving the comparisons of smokers and non-smokers with a greater degree of accuracy
other future projects
Other Future Projects
  • Analyze the amount of cyanide intake from certain foods and vitamins (B12) compared to tobacco smoke
  • Amount of cyanide in cigarette smoke compared to things such as vehicle exhaust, metal industry emissions, etc.
  • USF for the use of its instrumentation
  • Dr. Frank Pascoe, Dean of Arts and Sciences for his grant support
  • Alberto Juarez, USF graduate, for his work on phase I of this project
  • Dr. Salim M. Diab, Team supervisor
  • Galanti LM. Specificity of salivary thiocyanate as marker of cigarette smoking is not

affected by alimentary sources. Clin. Chem., 1997 Jan; 43(1):184-5.

  • Lahti M, Vilpo J, Hovinen J. Spectrophotometric determination of thiocyanate in

human saliva. J Chem Ed. 1999 Sept;76(9): 1281-3

  • Luepker RV, Pechacek TF, Murray DM, Johnson CA, Hund F, Jacobs DR. Saliva
  • Thiocyanate: a chemical indicator of cigarette smoking in adolescents. Am J
  • Public Health. 1981 Dec;71(12):1320-4.
  • O S Oluwole, A O Onabolu, I A Cotgreave, H Rosling, A Persson, and H Link
  • Incidence of endemic ataxic polyneuropathy and its relation to exposure to cyanide in a Nigerian communityJ. Neurol. Neurosurg. Psychiatry, Oct 2003; 74: 1417 - 1422.
  • White WLB, Arias-Garzon DI, McMahon JM, and Richard T. Sayre
  • Cyanogenesis in Cassava: The Role of Hydroxynitrile Lyase in Root Cyanide ProductionPlant Physiology, Apr 1998; 116: 1219 - 1225.
  • Wood John L. and Edward F. Williams, Jr.
  • http://www.acsu.buffalo.edu/~koudelka/kinetics/kineticsproblemset1answers.pdf
  • http://www.rxlist.com/cgi/generic3/nitroprusside_cp.htm