PDP 406 CLINICAL TOXICOLOGY

Pharm.D

Fourth Year
FOOD AND TOXICITY:
Natural Toxins

Mr.D.Raju.M.Pharm.,
Lecturer
 Hazards in food

• Physical: glass, stone, metal, wood, etc

• Chemical:
- natural toxins
- residues
- metals
- toxins formed during food processing
• Microbiological: pathogenic microorganisms
(bacteria, viruses, parasites, etc)
 Foodborne Diseases

Infections

Intoxications Toxicoinfection Invasive Infection

Chemical Poisonous Poisonous Microbial Other Neurotoxins Enterotoxins

Poisoning Plant Animal Intoxications
Tissues Tissues

Intestinal Systemic Other

Mucosa Tissues or
Organs
(Muscle,
Mycotoxins Algal Toxins Bacterial Liver, Joints,
(Fungal Toxins Fetus, Other)
Toxins)

Diarrhogenic Emetic Enterotoxins Neurotoxins Other

 Natural Toxins

• They are naturally present in plants & animals.

• The long term ingestion of natural toxins in
commonly eaten foods Æ the risks to human
health?
• Usually, natural toxins are not acutely toxic, except
in a few cases in animals.
• Most of the natural toxins, particularly those
occurring in plant-derived foods, induce adverse
effects only after chronic ingestion or by allergic
reactions.
 Risks of toxins in food

• General toxicity (instant death or illness)

• Carcinogenic
• Mutagenic
• Teratogenic
• Endocrine disrupters (hormones)
• Microbial pathogens
 Natural Toxins in Foods

• Endogenous toxins of plant origin

ƒ Toxic phenolic substances: flavonoids, tannins, coumarin,
safrole, and myristicin
ƒ Cyanogenic glycosides
ƒ Glucosinolates
ƒ Acetylcholinesterase inhibitors
ƒ Biogenic amines
ƒ Central stimulants
• Natural contaminants
ƒ Mixing of edible plants with toxic plants
ƒ Contamination resulting from intake of toxic substances by
animals
ƒ Microbial toxins
Endogenous toxins of plant origin
 Flavonoids

• A class of plant pigments that are widely present in

human food, are the flavonoids.
• These pigments are polyhydroxy-2-phenylbenzo-γ-
pyrone derivatives, occurring as aglycones,
glycosides and methyl ethers.
• A group of yellow pigments that occurs abundantly
is the flavones.
• Examples are nobiletin, tangeretin (in citrus fruits)
and 3, 3′, 4′, 5 ,6 ,7, 8-heptamethoxyflavone (in
grapefruit).
 Flavonoids

• The flavones are generally located in the oil vesicles

of the fruit peel. Flavones are apolar, and therefore
readily soluble in the oil.
• They can be found in the juice after pressing the
peel.
• The flavones group has been extensively
investigated for mutagenicity.
• A well-known mutagenic representative is quercetin,
occurring, for example, in cereal crops. Quercetin is
the only flavonoid shown to be carcinogenic in
mammals after oral administration.
 Tannins

• Tannins are a heterogeneous group of broadly

distributed substances of plant origin.
• Two types of tannins can be distinguished on the
basis of degradation behavior and botanical
distribution, namely hydrolyzable tannins and
condensed tannins.
• The hydrolyzable tannins are gallic, digallic, and
ellagic acid esters of glucose or quinic acid.
• An example of this group is tannic acid, also known
as gallotannic acid, gallotannin, or simply tannin.
Tannic acid has been reported to cause acute liver
injury, i.e., liver necrosis and fatty liver.
 Tannins

• The condensed tannins are flavonoids. They are

polymers of leukoanthocyanidins.
• Tannins occur in many tropical fruits, including
mango, dates, and persimmons.
• The contribution of the tannins in tea, coffee, and
cocoa to the total tannin intake by humans is of
particular importance. Tea has the highest tannin
content.
• Other important sources of tannins are grapes,
grape juice, and wines.
• A person may easily ingest 1 g or more tannins per
day.
 Safrole, coumarin, myristicin

• Coumarin widely occurs in a number of natural

flavorings, including cassis, lavender, and lovage.
These flavorings are extensively used in sweets and
liquors. Traces of coumarin occur in citrus oils and
some edible fruits.
• Safrole has been shown to cause liver tumors in
rats. It is found in the oil of sassafras and in black
peppers.
• Both coumarin and safrole are still allowed for food
use in the European Community. They are
prohibited in the US though, as they have been
found to cause liver damage in rats.
 Safrole, coumarin, myristicin

• Myristicin is found in spices and herbs such as

nutmeg, mace, black pepper, carrot, parsley, celery,
and dill.
• It has been suggested that myristicin contributes to
the toxicity of nutmeg. After nutmeg abuse,
tachycardia, failing salivation, and excitation of the
central nervous system have been reported. Nutmeg
has been abused as a narcotic.
 Cyanogenic glycosides

• Cyanogenic glycosides are glycosides from which

cyanide is formed by the activity of hydrolytic
enzymes. They are widely spread in higher plants.
• Can be found in food plants like cassava, lima
beans and the seed of some fruits (e.g. peaches) Æ
cyanide content.
• Fresh cassava cortex produces cyanide in quantities
ranging from 1.0 to more than 60.0 mg per 100 g,
depending on several conditions, including variety,
source, time of harvest and field conditions.
• Damaged roots can contain even more cyanide, i.e.,
245 g per 100 g.
Cyanogenic glycosides

Jansenn, Put & Nout (1997)

Cyanogenic glycosides

Jansenn, Put & Nout (1997)

 Cyanogenic glycosides

• It can be fatal if those foods are eaten raw or

prepared improperly.
• Peeling, washing in running water and cooking or
fermenting can remove and volatilize the cyanide.
• Consumption of cassava may lead to goiter, as the
cyanide formed can be metabolized to thiocyanate
by the enzyme rhodanase.
Goitrogens (Glucosinolates)

• Glucosinolates are a particular group of substances,

occurring in cruciferous plants, such as cabbage
and turnips. They can be considered as natural
toxins, but also as antinutritives.
• Presents in many commonly consumed plants, such
as cabbage, cauliflower, brussel sprouts, broccoli,
turnip, radish, oil seed meals.
• Inhibit the uptake of iodine by the thyroid Æiodine
deficiency.
• Concerning toxicity and antinutritive activity, the
hydrolysis products are the active agents, not the
glucosinolates themselves.
Goitrogens (Glucosinolates)

• Hydrolysis of glucosinolates results in the formation

of isothiocyanates and nitriles.
• The enzyme becomes available for catalysis when
cells are damaged on cutting or chewing.
• Several isothiocyanates have been shown to be
embryotoxic in rats, while in vitro studies have
proved a number of them to be cytotoxic and
mutagenic.
• Further, several nitriles have been identified as
precursors of N-nitroso compounds.
 Acetylcholinesterase inhibitors

• Acetylcholinesterase inhibitors have been detected

in several edible fruits and vegetables.
• Their active components are alkaloids.
• In potato, eggplant and tomato — members of the
Solanaceae family — the principal alkaloids have
been identified.
• The most potent inhibitors are found in potatoes,
and of these the most active component is the
glycoalkaloid solanine.
• Oral administration of solanine results primarily in
gastrointestinal and neurological symptoms.
 Acetylcholinesterase inhibitors

• The solanine concentration of potato tubers varies

with the degree of maturity at harvest, rate of
nitrogen fertilization, storage conditions, variety, and
greening by exposure to light.
• Commercial potatoes contain 2 to 15 mg of solanine
per 100 g fresh weight.
• Greening of potatoes may increase the solanine
content to 80 to 100 mg per 100 g.
• Most of the alkaloid is concentrated in the skin.
• Sprouts may contain lethal amounts of solanine.
• Peeling will remove most of solanine
 Acetylcholinesterase inhibitors

• Since potatoes also contain other glycoalkaloids,

namely chaconine and tomatine, with biological
properties similar to solanine, the symptoms seen in
potato poisoning may be due to combined actions of
the alkaloids.
• Solanine is heat stable and insoluble in water.
Hence, toxic potatoes cannot be rendered harmless
by cooking.
• It is generally accepted that 20 mg solanine per 100
g fresh weight is the upper safety limit.
 Biogenic amines

• Low molecular weight organic bases, which

pose biological activity.
• Naturally present in animals, plants and
microorganisms.
• Sources: fermented foods, vegetables, meats
• Mainly caused by the enzymatic decarboxylation
of amino acids by microorganism origin.
• High concentration in fish and cheese.
Biogenic amines

Jansenn, Put & Nout (1997)

Biogenic amines
Biogenic amines
 Biogenic amines

Factors influencing the formation of biogenic amines:

• Microorganisms:
Amine-producing bacteria - (enterococci, lactobacilli,
enterobacteriaceae, pediococci)
• pH
• Temperature
• Salt concentration (cheese)
• The availability of free amino acids
• The level of starter culture (cheese)
 Biogenic amines

• Legal upper limit (Silla-Santos, 1996):

- histamine: 100 mg/kg food
- tyramine: 100-800 mg/kg food
- phenyletylamine: 30 mg/kg food
• 1000 mg/kg amine (based on histamine intoxication
& amine concentration in food) is dangerous for
human health.
• The toxicity dose of BA depends on the individual
sensitivity and characteristic.
 Biogenic amines

• (+) - cell proliferation

- regulation of nucleic acid function
- protein synthesis

• (-) - histamine intoxication

- nausea
- respiratory distress
- hot flushes
- heart palpitation
- headache
- hypertension & hypotension
 Mushroom Toxins

• Caused by the high content of amatoxins in

mushrooms.
• Mushrooms identified as containing amatoxin
toxins are the species Amanita bisporigera, A.
temifolia, A. ocreata, A. suballiacea, Galerina
autumnalis, and Lipiota brunneolilacea.
 Mushroom Toxins

• There are four categories of mushroom toxins:

1. Neurotoxins
Cause neurological symptoms such as profuse
sweating, hallucinations, depression, spastic
colon, excitement, convulsions, and coma.

2. Protoplasmic poisons
Cause generalized destruction of cells, which is
followed by organ failure.
 Mushroom Toxins

3. Gastrointestinal irritants
Produce rapid, transient nausea, abdominal
cramping, vomiting, and diarrhea.
4. Disulfram-like toxins
Disulfram-like toxins are usually nontoxic and
produce no symptoms. However, if alcohol is
consumed within 72 hours after eating them,
they may produce vomiting, nausea,
headache, flushing, and cardiovascular
disturbances.
 Mushroom Toxins

• The first symptoms of mushroom poisoning

occur within 6–24 hours after ingestion of the
mushrooms (phase one).
• Phase two, also called the gastrointestinal
phase, involves severe vomiting and abdominal
cramps, nausea, and watery diarrhea.
• Phase three lasts about 12–24 hours and is
characterized by improved clinical symptoms;
however, it is also the beginning of liver
necrosis.
 Mushroom Toxins

• Phase four (the last phase), results in hepatic

failure, encephalopathy, internal bleeding, and,
often, acute renal failure. Internal bleeding is
usually observed and may cause complications
and death.
• Patients usually die within 5–20 days after
ingestion of the mushrooms.
Natural Contaminants
• Natural contaminants can also originate from biological
systems different from those in which they occur.
• There are three important sources:
ƒ Raw materials of plant origin can become
contaminated if they are mixed with toxic non-nutritive
plant species.
ƒ Raw materials of animal origin, mainly fish and milk,
can also become contaminated if the animal has
ingested toxic substances of natural origin.
ƒ Contaminants of natural origin can be the products of
microorganisms.
Mixing of edible plants with toxic plants

• Several intoxications have been reported following

the consumption of contaminated cereals.
• The causative agents are pyrrolizidine alkaloids,
produced by the genera Senecio, Crotalaria and
Heliotropium.
• Pyrrolizidine alkaloids can be the cause of acute
liver damage and vein lesions. These substances
may also contribute to the liver cancer incidence in
humans.
Contamination resulting from the intake of toxic substances
by animals: Natural toxins in aquatic organisms

• Paralytic shellfish poisoning (PSP) is attributed to

the consumption of shellfish that have become
contaminated with a toxin or group of toxins from the
ingestion of toxic plankton, in particular toxic
dinoflagellates.
• The shellfish involved are pelecypods, a family of
mollusks, including mussels and clams.
• The dinoflagellates produce a complex mixture of
toxins. One of the components has been identified
as saxitoxin.
Contamination resulting from the intake of toxic substances
by animals: Natural toxins in aquatic organisms

• Shellfish poisoning symptoms include tingling and

burning in face, lips, tongue, and ultimately the whole
body, and parathesia followed by numbness, general
motor incoordination, confusion, and headache.
• These symptoms develop within 30 minutes after
ingestion. Death, preceded by respiratory paralysis,
occurs within 12 hours.
• The chance of contamination and poisoning is highest
during a so-called red tide (dinoflagellate bloom) Æ
although the bloom may also be yellowish,brownish,
greenish, and bluish in color. The red color is probably
due to the xanthophyll peridinin.
Contamination resulting from the intake of toxic substances
by animals: Natural toxins in aquatic organisms

• The high toxicity of the paralytic shellfish poisons,

intoxication rarely occurs.
• This is largely due to strict regulations set by many
countries and the awareness in coastal areas of the
risks associated with eating shellfish during red
tides.
• Although ordinary cooking destroys up to 70% of the
toxin(s) and pan-frying destroys even more, there
may be sufficient toxin left in the mollusks to cause
serious poisoning.
 Microbial Toxin: Mycotoxin

• Mycotoxins are secondary metabolites of fungi

which can induce acute as well as chronic toxic
effects (i.e., carcinogenicity, mutagenicity,
teratogenicity, and estrogenic effects) in animals
and man.
• Currently, a few hundred mycotoxins are known,
often produced by the genera Aspergillus,
Penicillium, and Fusarium.
• Toxic syndromes resulting from the intake of
mycotoxins by man and animals are known as
mycotoxicoses.
 Microbial Toxin: Mycotoxin

• Mycotoxin contamination of food and feed highly

depends on the environmental conditions that lead
to mold growth and toxin production.
• The detectable presence of live molds in food,
therefore, does not automatically indicate that
mycotoxins have been formed.
• On the other hand, the absence of viable molds in
foods does not necessarily mean there are no
mycotoxins. The latter could have been formed at
an earlier stage prior to food processing.
• Because of their chemical stability, several
mycotoxins persist during food processing, while the
molds are killed.
Aflatoxins

• Aflatoxins are the most important mycotoxins, which

is produced by certain species of Aspergillus (A.
flavus and A. parasiticus), which develop at high
temperatures and humidity levels.
• Aflatoxins are carcinogenic substances and may be
present in a large number of foods. This toxin can
cause cancer, cirrhosis of the liver.
• For substances of this type there is no threshold
below which no harmful effect is observed.
Aflatoxins

• The most common commodities contaminated

are tree nuts, peanuts, and corn and cottonseed
oil.
• The major aflatoxins of concern are B1, B2, G1,
and G2 Æ usually found together in various
proportions. Aflatoxin B is usually predominant,
and it is the most toxic and carcinogenic.
• TLC method can detect aflatoxins.
Aflatoxins

• The aflatoxins B1, B2 fluoresce blue and G1, G2

fluoresce green when viewed under a
microscope.
• A major metabolic product of aflatoxin B1 is
aflatoxin M (usually excreted in the milk of dairy
cattle that have consumed aflatoxin-
contaminated feed).
• LD50 ranges from 0.5 to 10 mg/kg body weight
(any species of animals)
What are some of the regulations guiding
the levels of toxins in food?
• Commission Regulation (EC) No 466/2001 of 8
March 2001
Setting maximum levels of certain contaminants
in foodstuffs.
• USFDA
• CODEX STAN 193-1995 (Rev.1-1997)
How to establish the maximum limit of toxins or
contaminants in foods.