Date/Time : 22 September 2021/ 11.30 –14.

00
Lecturer :Drh Diah Nugrahani Pristihadi,
MSi
Group :2

CYANIDE

Group Members:

Nur Nadhirah Bt Rahmat B04188015

Yvonne Lee Yong Ai B04188021
Divheeya A/P Pathmanathan B04188025
Hana Shady Hidayah Sugeng B04188030
Hanifati Husna B04188031

Pharmacology and Toxicology Division

Department of Anatomy, Physiology and Pharmacology
Faculty of Veterinary Medicine
Bogor Agricultural University
2021
 Introduction

Cyanide is a chemical compound that contains the group C≡N. this group
known as the cyano group consist of a carbon atom triple-bonded to a nitrogen
atom. In inorganic cyanides, the group is present as the anion CN-. soluble salts
such as sodium cyanide and potassium cyanide are highly toxic. Hydrocyanic acid
also known as hydrogen cyanide or HCN is a highly volatile liquid that is
produced on a large scale industrially. It is obtained by acidification of cyanide
salts. Cyanide is produced by certain bacteria, fungi and algae. It is and
antifeedant in a number of plants. Cyanides are found in substantial amounts in
certain seeds and fruit stones, eg: bitter almonds, apricots, apples and peaches.
Chemical compounds that can release cyanide are known as cyanogenic
compounds. In plants, cyanides are usually bound to sugar molecules in the form
of cyanogenic glycosides and defend the plant against herbivores. Cassava roots
and important potato like food grown in tropical countries also contain
cyanogenic glycosides.

Literature review

Cassava has played important role in Indonesian diet in the past and will
continue to do so in the future. Human consumption of cassava and cassava
products is widespread in all parts of the country in both rural and urban areas,
and by most income classes (Dixon 1979). The cyanide anion is an inhibitor of
the enzyme cytochrome c oxidase (also known as aa3), the fourth complex of the
electron transport chain found in the inner membrane of the mitochondria of
eukaryotic cells. It attaches to the iron within this protein. The binding of cyanide
to this enzyme prevents transport of electrons from cytochrome c to oxygen. As a
result, the electron transport chain is disrupted, meaning that the cell can no
longer aerobically produce ATP for energy. Tissues that depend highly on aerobic
respiration, such as the central nervous system and the heart, are particularly
affected. This is an example of histotoxic hypoxia. Cyanide can enter your body if
you breathe air, eat food, or drink water that contains it. Cyanide can enter
through skin for people who work in cyanide related industries. Once it is in your
lungs or stomach, cyanide can quickly enter bloodstream. Some of the cyanide is
changed to thiocyanate which is less harmful and leaves body in the urine. A
small amount of cyanide is converted in the body to carbon dioxide which leaves
the body by exhale. At low levels of exposure to cyanide compounds most of the
cyanide and its products leave body within first 24 hours after exposure
(Anonymous 2013).
 Aim

The aim of this experiment is to determine the clinical symptoms of

cyanide poisoning and the antidote effects of sodium nitrite and sodium
thiosulfate tested on rabbits, identification of cyanide in plant samples and NaCN
solution using picrate paper and identification of cyanide in animal samples.

Materials and Tools

The tools and materials used were syringe, test tube, mortar, water bath,
picrat paper, test tube clamp, test tube plug, rabbit, 1% NaCN, NaOH, 1%
NaNO2, FeSO4, FeCl3, HCl, 5% Na2S2O3, aquadest and cassava leaves.

Procedure
A. Identification of CN in plants
Tube 1 was filled with aquadest as negative control (-), tube 2 NaCN + HCl
positive control (+) and tube 3 scour cassava leaves. Picrat was placed on the
plug, then it was heated. Discoloration was observed on the pictorial paper.

B. Identification of CN from samples of animal origin

1% NaCN solution was entered in the test tube then 1 mL 50% NaOH, 3 drops
FeSO4, and 3 drops FeCl3 was added. The tube was heated, then concentrated
HCl was added. The color change to blue berlin (Prussian Blue) was watched.

C. Clinical symptoms of cyanide and antidote poisoning

Rabbit was weighed. 1% NaNO2 solution and 5% Na2S2O3 solution were
prepared respectively as much as 2.5 mL in a different fluid. 5-10 mg / kg BW of
1% NaCN solution is administered by mouth; the rabbit uses the syringe that has
removed the needle. The clinical symptoms that occur in rabbits are noticed and
then injected the antidote intravenously through the auricular vein. The antidota
administration was started with 1% NaNO2 solution and then the solution
Na2S2O3 5%.

Results

Table 1. Identification of cyanide from plant sample

No. Test tube Color change of picrate paper

 1 Aquades (negative control) No change

2 Topioca Leaves Yellow to brick red

3 NaCN 1% + HCl 5% (positive control) Yellow to brick red

Table 2. Identification of cyanide in animal sample

Test tube Color change

NaCN 1% + NaOH 50% + FeSO4 10% + Brick red to prussian blue

FeCl3 10% + HCl (after heating)

Table 3. Observation of clinical symptoms of cyanide poisoning in rabbit

Time Clinical symptoms

0 minute Breathing rapid, pupil dilate

10 minute Breathing rapid, pupil dilate, incactive

After giving antidota (NaNO2 Active, pupil back to normal, breathing back
& Na2S2O3) to normal.

Discussion

Cyanide has a high binding affinity for the ferric ion on the cytochrome a3
portion of the enzyme. This effectively stops electron transport and inhibits
oxidative phosphorylation. This inhibition in essence terminates the synthesis
pathway of adenosine triphosphate. The mitochondria continue to be exposed to
an adequate oxygen supply; however, there is impaired oxygen extraction and use.
This disruption of aerobic metabolism leads to increased glycolysis by anaerobic
pathways and central nervous system and cardiovascular impairment. (Gracia and
Shepherd, 2004)
The toxic properties associated with fresh cassava leaves are due to the
free HCN that is liberated when their cyanogenic glucosides, namely linamarin
and lotaustralin, are hydrolysed. The release of free HCN is brought about by the
action of either the endogenous enzyme linamarase in damaged plant tissues or
β-glucosidases within the digestive tract of animals. The linamarase and
glucosides do not come into contact in healthy cassava leaves, but contact occurs
when the tissues are mechanically damaged or when the physiological integrity is
lost as in the case of wilted leaves (Ravindran 1993). The picrate paper method
for cyanide determination relies on endogenous enzymes to breakdown the
cyanogenic glucosides (linamarin) in cassava samples. The hydrogen cyanide
liberated in the process reacts with picrate papers that have a colour pigment
which darkens with released cyanide. This determines that the cassava leaf sample
used in practice contains cyanide due to indication of color change of picrate
paper from yellow to brick red.
The second experiment is determining cyanide content in animal products
indicated by the change and reaction of animal products with NaCN. Sodium
cyanide was tested against water and the test tube with sodium cyanide turned
blue upon adding HCl. This is due to the formation of Prussian Blue, which
indicates the presence of the ferric-hexacyanoferrate, Fe4[Fe(CN)6]3. The
incubation pH and exposure time, play major roles in the cyanide release from
Prussian Blue. Longer exposure times at a lower pH are known to release higher
amounts of cyanide. (Yang et al. 2007)
The last experiment is the observation of clinical symptoms of cyanide
poisoning in rabbits. Cyaninde was orally administered. Rabbit was observed to
be inactive at minute 10. Rabbit also has rapid breathing and its pupils are dilated.
According to Hamel (2011), the onset of signs and symptoms is usually less than
1 minute after inhalation and within a few minutes after ingestion. Early
respiratory signs of cyanide poisoning include transient rapid and deep
respirations. This respiratory change reflects stimulation of peripheral and central
chemoreceptors within the brain stem in an attempt to overcome tissue hypoxia.
Antidotes used for treating cyanide poisoning are sodium nitrite (NaSO2)
and sodium thiosulfate (Na2S2O3). Nitrites reduce blood cyanide levels by causing
the formation of methemoglobin, to which cyanide binds with higher affinity than
it does to cytochrome oxidase. Binding of cyanide to methemoglobin liberates
cytochrome oxidase, which is necessary for aerobic cellular respiration (Hall
2009). Antidote was administered via vena auricularis. Rabbit gained
consciousness and began to be active seconds after administration. Pupil sizes are
back to normal and breathing is also coordinated.

Conclusion

In conclusion, cyanide toxicity is rapid acting and can cause central

nervous and respiratory problems. Cassava leaves can contain an amount of
cyanide that can cause poisoning if consumed. Animal products which contain
cyanide turned blue when adding HCL causing the formation of (Fe4(Fe(CN)6)3.
Cyanide toxicity can be reduced with antidotes containing nitrate such as sodium
nitrate or sodium thiosulfate.
 References

Anonymous. 2013. Toxicological Profile for Cyanide. Georgia (US): U.S.

Department of Health and Human Services, Public Health Service,
Agency for Toxic Substances and Disease Registry.

Dixon J. 1979. Production and consumption of cassava in Indonesia. Bulletin

of Indonesia Economic Studies. 15(3): 83-106.

Gracia R, & Shepherd G. 2004. Cyanide Poisoning and Its Treatment.

Pharmacotherapy, 24(10), 1358–1365.

Hall AH, Saiers J, & Baud F. 2009. Which cyanide antidote? Critical Reviews in
Toxicology, 39(7), 541–552.

Hamel J. 2011. A Review of Acute Cyanide Poisoning With a Treatment Update.

Critical Care Nurse, 31(1), 72–82.

Ravindran V. 1993. Cassava leaves as animal feed: Potential and limitations.

Journal of the Science of Food and Agriculture, 61(2), 141–150.

Yang Y, Brownell C, Sadrieh N, May J, Del Grosso A, Place D, … Faustino P.

2007. Quantitative measurement of cyanide released from Prussian Blue.
Clinical Toxicology, 45(7), 776–781.