CYANOGENIC

GLYCOSIDES
BY
PHARM. EVANS PAUL KWAME AMEADE
DEPT. OF PHARMACOGNOSY AND HERBAL MEDICINE
SCHOOL OF PHARMACY AND PHARMACEUTICAL SCIENCES
UNIVERSITY FOR DEVELOPMENT STUDIES
PHARM.D, LEVEL 300
JANUARY, 2023
Outline
• What are cyanogenic glycosides
• Sources of cyanogenic glycosides
• Structure of Cyanogenic glycosides
• Processes of hydrogen cyanide production
• Biogenesis of cyanogenic glycosides
• Importance of cyanogenic glycosides to the life
• Effects of cyanogenic glycosides on animal and human
• Symptoms of cyanide poisoning
• Management of poisoned animals
• Effect of processing on cyanogenic glycosides in food
• Test for cyanogenic glycosides
What are Cyanogenic glycosides?
• Cyanogenic glycosides are a group of plant secondary
metabolites in plants that releases hydrogen cyanide
when chewed or digested through a process called
cyanogenesis.
• Hydrogen cyanide (HCN) is a colorless, rapidly acting,
highly poisonous gas or liquid that has an odor of
bitter almonds
• A solution of hydrogen cyanide in water is called
hydrocyanic acid or prussic acid.
Sources of cyanogenic glycosides
• Cyanogenic glycosides occur in more than 3000 plant
species from 130 families including inter alia the
following;
• Euphorbiaceae
• Rosaceae
• Asteraceae
• Passifloraceae
• Poaceae
• Plant species used as food or forage known to contain CG
include:
• Cassava (Manihot esculenta) - linamarin, while small
amount of lotaustralin (methyl linamarin) and amygdalin
• Linseed (Linum usitatissmium) - linustatin, neolinustatin
and linamarin
• Sorghums (Sorghum spp)– dhurrin, amygdalin
• White clover (Trifolium repens) - linamarin and lotaustralin
• Some species of Lotus - lotusin
• Yam (Bitter yam) - Diosgenin
• Cocoyam - Amygdalin
• Bamboo shoots – Taxiphyllin
• Beans such as Phaseolus vulgaris - Linamarin
• Kernels of
• almonds (Amygdalus communis) - Amygdalin
• apricots (Prunus armeniaca – bitter type) - Amygdalin and
prunasin
• peaches (Prunus persica)- Amygdalin and prunasin
• apples (Malus sylvestris) - Amygdalin
Apple Cassava Linseed Sorghum

Lotus Bamboo shoots Bitter yam Almond

Apricot Peaches
Structure of cyanogenic glycosides
• There more than 75 different cyanogenic glycosides
• They are typically O-β-glycoside of α-hydroxynitriles (cyanohydrins)
• Nitrile is any organic compound that has a −C≡N functional group
• A generic structure for the cyanogenic glycosides is as below, where R
is variable. R1 is either an aliphatic or aromatic group and R2
is mainly H
O-linkage

Glycone unit

Nitrile
• Linamarin and lotaustralin are the two most commonly
found cyanogenic glycosides and present in many plant
families
• Other notable ones include amygdalin in Rosaceae and
dhurrin found in poaceae families.
• The sugar portion of the molecule may be a
monosaccharide or a disaccharide such as gentiobiose
or vicianose.
• If the glycone is a disaccharide, enzymes present in
the plant may bring about hydrolysis in two stages, as
in the case of amygdalin
• In all cases the first sugar attached to the aglycone is
β-D-glucose
Processes of Hydrogen cyanide production
• The hydrogen cyanide does not exist freely in the plant
• HCN is produced only when plant tissues are damaged by herbivory,
or when they are disintegrated during processes, such as grinding,
pounding or in the presence of water for example during soaking or
fermentation
• The enzyme, β-glycosidases that are kept in other tissues separate
from cyanogenic glycosides are released to contact with the
glycoside.
• The β-glycosidases breaks apart the glycosidic bond in the cyanogenic
glycoside to release sugars and cyanohydrin (hydroxynitriles)
• The cyanohydrins are degraded spontaneously but slowly by
a second enzyme, hydroxynitrile lyase, into hydrogen cyanide
and ketones (or aldehydes)
• Amygdalin is hydrolyzed by β-glucosidase to release
gentiobiose and L-mandelonitrile.
• Gentiobiose is further hydrolyzed to give glucose, whereas
mandelonitrile (the cyanohydrin of benzaldehyde)
decomposes to give benzaldehyde and hydrogen cyanide.
• To prevent unwanted production of hydrogen cyanide in the
plant, the cyanogenic glycosides and the hydrolytic enzymes
are in different parts of the plant or cells. Eg.
• Shoot of sorghum, Sorghum bicolor
• Glycoside dhurrin (vacuoles of epidermal cell)
• β-glucosidase (chloroplasts of the mesophyll cell)
• Hydroxynitrile lyase (cytosol of the mesophyll cells)
• Cassava
• Glycosides linamarin and lotaustralin (widely distributed among
tissues)
• Linamarase (concentrated in laticifer and latex)
• Hydroxynitrile lyase (concentrated in the cell wall of leaf tissues
but absent in the root)
Biogenesis of Cyanogenic glycosides
• Cyanogenic glycosides are derived from six amino acids namely
• L-valine
• L-isoleucine
L-leucine
• L-phenylalanine
• L-tyrosine
• A non protein amino acid cyclopentenyl-glycine
• These amino acids are precursors for the production of the aglycone
portion of the cyanogenic glycoside through intermediate metabolites
such as N-hydroxyamino acids, aldoximes, nitriles and α-
hydroxynitriles.
• Cyanogenic glycoside biosynthetic pathways can be classified
into three stages
• Step- I: A precursor amino acid is converted to aldoxime through two
successive N-hydroxylation of amino group of parent amino acid by
an enzyme of cytochrome-P450 family.
• Step- II: aldoxime converted to cyanohydrin using another
cytochrome-P450 enzyme.
• Step- III: cyanohydrins get glycosylated by a soluble enzyme
UDP-glucosyltransferase
• Levels of cyanogenic glycosides are highest in actively growing
parts of the plant and are increased under conditions of
• Stress
• high soil nitrogen
• low soil phosphorus
General scheme of the biosynthesis of cyanogenic glycoside in plants
Biosynthesis of linamarin and lotaustralin in Lotus japonicus
• Some cyanogenic glycosides and their amino acid precursors
are as follows;
• Valine and isoleucine:
• linamarin and lotaustralin
• Leucine:
• sutherlandin, epidermin, osmaronin, dihydro-osmaronin and
epiheterodendrin have been reported from the barley (H. vulgare)
seedlings
• L-tyrosine:
• dhurrin, triglochinin, taxiphyllin, holocalin and vicianin
• L-phenylalanine:
• prunasin (an aromatic cyanogenic glycoside), acacipetalin,
proacacipetalin, 3-hydroxyheterodendrin, sambunigrin, amygdalin
• Cyclopentenyl-glycine:
• Dhurrin, taxiphyllin, prunasin, sambunigrin, deidaclin and tetraphyllin A
Importance of cyanogenic glycosides to the life
In plants, the following are some roles of cyanogenic glycosides
• As hydrogen cyanide is highly toxic to most animals and insects, cyanide
glycoside-borne defense systems are effective against herbivory.
• Some studies showed the possibility of cyanogenic glycosides serving as
nitrogen storage compounds.
• It is possibly a way of storage of reduced nitrogen and sugar
• Improve plant plasticity, i.e., establishment, robustness, and viability in
response to environmental challenges
• Cyanogenic glucosides may be involved in modulating oxidative stress by
scavenging reactive oxygen species
Some studies have shown beneficial role of cyanogenic glycosides in animals.
• Cyanogenic glycosides from Prunus spp (cherry), particularly amygdalin (sold
under the trade name Laetrile) and prunasin have shown anticarcinogenic
activity in vitro studies.
Effects of cyanogenic glycosides on animal and human
• There are reported deaths in animals after consuming forages with high
cyanogenic glycosides
• The toxicity of cyanogenic glycosides and their derivatives is dependent on
the release of hydrogen cyanide.
• Toxicity may result in acute cyanide poisoning and has also been implicated
in the etiology of several chronic disease especially in Africa and South
America where cassava is their staple food.
• Konzo
• Tropical ataxic neuropathy
• Goitre
• Tropical amblyopia
• Konzo (upper motor neuron disease): irreversible but
nonprogressive symmetric spastic paraparesis that has an abrupt
onset occurring mostly in children and women of childbearing age
• Tropical ataxic neuropathy: sore tongue, optical atrophy,
neurosensory deafness, and sensory gait ataxia
• Goitre occurs in continuous exposure to dietary cyanide by the
interferences of thiocyanate (the end products of cyanide
detoxification in human system which is a goitrogen) with dietary
iodine, thus leading to iodine deficiency
• Tropical amblyopia characterized by atrophy of the optic nerve,
resulting in blindness.
Symptoms of cyanide poisoning
• As cyanide is extremely toxic, one of the most obvious symptoms is death.
• In the body, cyanide acts by inhibiting cytochrome oxidase, the final step
in electron transport, and thus blocks ATP synthesis.
• Prior to death, symptoms include
• Headache
• Dizziness
• Mental confusion
• Faster and deeper respiration,
• Faster irregular and weaker pulse,
• Salivation and frothing at the mouth,
• Muscular spasms
• Dilation of the pupils
• Bright red mucous membranes
• When an animal consumes forage releasing toxic levels of HCN,
clinical signs becoming evident within 30 minutes of ingestion.
• Animals exhibit excitement, tremors, rapid respiration, tachycardia,
dyspnea, collapse, terminal convulsions, coma, and death.
• Blood and tissues are a characteristic cherry-red because the
hemoglobin is oxygenated but cells cannot take up the oxygen. e
• The blood clots slowly or not at all. Hemorrhages are often present
under the epicardium and endocardium
Management of poisoned animals
• Poisoned animals should be treated with intravenous sodium nitrite
(22 mg/kg) and sodium thiosulfate (600 mg/kg).
• These oxidize hemoglobin (Fe +2) , forming methemoglobin (Fe+3),
which binds cyanide and protects the cytochrome oxidase system
from its effects.
• The best treatment is prevention – avoiding harvesting and feeding
these plants when they are likely to be toxic.
Effect of processing on cyanogenic glycosides in
food
• Processing methods, such as peeling, drying, grinding,
soaking, boiling or cooking, soaking and fermentation have
been reported by several studies to cause significant
reduction in the cyanogenic glycosides of processed foods.
• Food‐processing methods generally disintegrate cyanogens
contents of plants, and this leads to the production of
cyanide.
• Since cyanide is volatile, further processing techniques, such
as roasting and drying, will volatilize the remaining cyanide
to low level
Test for cyanogenic glycosides
• Picrate-impregnated paper test (According to Harbone, 1972)
• The test should be performed in triplicate.
• Fresh plant material was cut into small pieces and placed in a test tube with 1.5mL of
distilled water, and 6 drops of chloroform, followed by briefly crushing the material with a
glass rod.
• The tube was stoppered with a cork containing a strip of picrate-impregnated paper
hanging down from the stopper, and incubated at ambient temperature for 2 h.
• A colour change of the paper, from yellow to brown-red, indicated the release of HCN by
the plant.
• If there was no release of HCN within 2 h, indicating a negative test, the tube was left at
ambient temperature for 24 and 48 h, so that it could be re-examined.
• A brown-red coloration within 2 h indicated the presence of cyanogenic glycoside and the
respective hydrolytic enzyme, and the plants were considered cyanogenic in the field.
• A brown-red color appearing within 48 h indicated that the cyanogenic glycoside
spontaneously released HCN without the action of enzyme.
• No colour change after 48 h indicated that the test was negative for cyanogenic glycoside.
• Ferriferrocyanide test:
• Macerate 1 g of the powdered drug with 5 ml of alcoholic KOH for 5
min.
• Transfer it to an aqueous solution containing FeS04 and FeCl3, and
maintain at 60-70°C for 10 minutes.
• Now transfer the contents to HCl (20%) when the appearance of a
distinct Prussian blue colour confirms the presence of HCN.
References
• https://www.sciencedirect.com/topics/biochemistry-genetics-and-mo
lecular-biology/cyanogenic-glycosides
• https://www.sciencedirect.com/topics/biochemistry-genetics-and-mo
lecular-biology/prunasin
• http://www.plantphysiol.org/content/plantphysiol/94/2/401.full.pdf
• Gleadow, R. M., & Møller, B. L. (2014). Cyanogenic glycosides:
synthesis, physiology, and phenotypic plasticity. Annual review of
plant biology, 65, 155-185.
• https://www.sciencedirect.com/topics/agricultural-and-biological-scie
nces/cyanogenic-glycosides
THANKS