FAO Consumer Protection Fact Sheets

No.2: Pyrrolizidine Alkaloids

PYRROLIZIDINE ALKALOIDS IN FOODS AND

ANIMAL FEEDS
WHAT ARE PYRROLIZIDINE ALKALOIDS?
Pyrrolizidine alkaloids (PAs) are toxins found naturally in a wide variety of plant species. PAs
are probably the most widely distributed natural toxins and affect wildlife, livestock and
humans. Over 6 000 plant species are known to contain PAs, although direct poisonings in man
and animals seems to be associated with only a few species. Poisoning caused by these toxins is
associated with acute and chronic liver damage and is commonly fatal.

Direct human cases of poisoning are occasional but are well-documented; consumption of grain
or grain products (flour or bread) contaminated with seeds from weed species that contain
these alkaloids is commonly involved. These occurrences frequently occur as “outbreaks”
following dry season or drought conditions that favour the development of weeds in the
primary crop. It has been suggested that intoxication may occur as a result of drinking milk
from affected animals, but amount of PAs expressed in the milk of animals exposed to PAs is low
(0.4-0.8% of the ingested dose). The same holds true for eggs. The ingestion of honey from
bees that have fed on toxic plant species has also been suggested as a possible exposure
pathway, but again, the levels involved are low and there are no reported cases. The direct and
deliberate use of toxic plant species as herbal teas or traditional medicines1 forms another
pathway of exposure which is well-documented and has resulted in deaths.

Farm animals, particularly cattle, sheep, goats, horses, poultry and pigs are known to be
susceptible to poisoning with high levels of mortality, while small animals such as rabbits
appear less affected. There are reports of toxicity to fish. Outbreaks in farm animals cause
severe economic losses to farmers and rural communities and, as noted above, there is the
possibility of transfer of the toxins to humans.

OCCURRENCE
Over 350 PAs are known and the list continues to grow. They are known to be present in more
than 6 000 plant species. The main sources are the families Boraginaceae (all genera),
Compositae (tribes Senecionae and Eupatoriae), and Leguminosae (genus Crotalaria). Some
plant species express several PAs or alkaloid N-oxides and there are some PAs that are
expressed by several plant species. The toxins are commonly concentrated in the seeds and the
flowering parts of the plant, with decreasing amounts in the leaves, stems and roots. Most plants
produce mixtures of PAs in varying concentrations ranging from less than 0.001 % to 5 % in
certain plant seeds. Some PA-bearing plants are used as ground cover, soil improvers
(Leguminosae), ornamental plants, and for animal feed. Some, especially in the Boraginaceae
family, are appreciated for the quality of their honey.

1 Herbal preparations and medicines are considered as therapeutic goods (i.e., not as foods) for the
purpose of this Fact Sheet. Nevertheless, some of the human data on exposure to PAs is based on this use
and will be referred to as required.

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Excluding the use of herbal teas and medicines, the plants most commonly reported as being
associated with food poisoning in humans are Heliotropium (in the family Boraginaceae ) and
Crotolaria. These occur as weeds in cereal or legume crops and the seeds are mingled
accidentally with the main crop at harvest. The toxins survive the milling, baking and
subsequent processes. The situation is frequently aggravated by drought and other conditions
advantageous to weed growth at the expense of the crop. Outbreaks of veno-occlusive disease
and other liver disorders have been reported from parts of Central Asia, Afghanistan and India
with recent outbreaks occurring in Afghanistan (International Programme on Chemical Safety,
1988), (WHO, 2001), (Anonymous, 2001).

Poisoning in animals has been reported from all of the sources listed above with known
outbreaks attributed to Heliotropium, Trichodesma, Senecio, and Crotalaria species. In general,
grazing animals will avoid PA-bearing plants but may have little choice in conditions of drought
or when searching for food on over-grazed or otherwise depleted pastures. If weedy crops are
used for the production of hay or silage the animals can no longer exercise discrimination when
feeding because the toxins survive storage processes and are completely intermingled with the
fodder. Mortality is reported to be high.

CHEMISTRY
PAs are heterocyclic compounds and most of them are derived from four necine bases:
platynecine, retronecine, heliotridine and otonecine. Retronecine and heliotridine are
enantiomers at the C7 position. Most of the naturally occurring PAs in plants are esterified
necines or alkaloid N-oxides (except for the otonecine-type alkaloids), whereas unesterified PAs
hardly occur in plants. The esters can be divided in monoesters, non-macrocyclic diesters and
macrocyclic diesters of a necine base. Figure 1 shows the basic structure of the four necine
bases forming toxic PAs.

a. b. c. d.

Figure 1: Basic structure of the four necine bases forming toxic PAs; a. platynecine, b. Retro-
necine, c. heliotridine, d. otonecine (European Food Safety Authority, 2007).

The minimum structural requirements for toxicity are:

1. an unsaturated 3-pyrroline ring;
2. one or preferably two hydroxyl groups, each attached to the pyrroline ring via one
carbon atom;
3. at least one of the hydroxyls is esterified;
4. the acid moiety has a branched chain (Australia New Zealand Food Authority, 2001).

TOXICITY

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The toxicology of PAs was reviewed comprehensively by the International Programme on

Chemical Safety in 1988 (International Programme on Chemical Safety, 1988). Updated reviews
are available from the Australia New Zealand Food Safety Authority (Australia New Zealand
Food Authority, 2001) and the European Food Safety Authority (European Food Safety
Authority, 2007).

Poisoning by PAs is due to in acute and chronic liver damage. In affected animals and humans,
PAs demonstrate marked toxicity to the liver, resulting in hepatocellular injury, cirrhosis and
veno-occlusive disease. In extreme cases neighbouring organs such as the heart and lungs may
be affected. Symptoms are those of liver failure and cirrhosis. In humans, it is reported that
following a poisoning outbreak in which sub-acute toxicity is observed, some 50% of patients
will recover completely and 20% will die rapidly. Of the survivors about 20% will appear to
recover clinically but may go on to develop cirrhosis and liver damage in later years. (K.L.
Stuart , G. Bras, 1957). In animals, symptoms have been described as “depression, photo-
sensitization, scouring, straining, depraved appetite, staggering gait, circling and death” (North
West Weeds, 2007).

Although metabolites of PAs have been shown to have mutagenic activity, mainly in Drosophila
and many have been shown to be carcinogenic, mainly in the rat, there is no evidence of
pyrrolizidine alkaloid-induced cancer in humans.

The Australia New Zealand Food Authority has estimated a provisional tolerable daily intake
(PTDI) for PAs in humans of 1 µg/kg body weight/day. This was calculated on the basis of veno-
occlusive disease of the liver as the major toxicological effect of chronic exposure. According to
the Authority, “the available data on cases of veno-occlusive disease in humans indicate that a
tentative no-observed-effect level (NOEL) of 10 μg/kg bw/day can be established. If an
uncertainty factor of 10 to account for human variability is applied to this NOEL, the provisional
tolerable daily intake (PTDI) for PAs in humans is 1 μg/kg bw/day” (Australia New Zealand
Food Authority, 2001).

METHODS OF ANALYSIS
Methods of analysis for PAs in animal feeds have recently been reviewed by the European Food
Safety Authority (European Food Safety Authority, 2007). The Authority notes that various
analytical techniques, particularly chromatographic methods in conjunction with mass
spectrometry can be used to detect PAs in plants or plant derived products. None of these
methods, however, has been validated for the analysis of (mixed) feed samples. It also notes on-
going research into the use of ELISA techniques.

Methods for the detection of PAs in biological fluids (milk, blood plasma, bile, urine) and also in
honey were also reviewed by the Authority.

There are no official methods for the detection or determination of PAs in foods and there is
apparently no systematic analysis for PAs in grains entering the food supply (Australia New
Zealand Food Authority, 2001). A useful review of screening methods and confirmation assay
development for plant-assocoated toxins in animal feed is provided by Than et al. (K.A. Than,
2005)

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A method for the determination of toxic seeds in samples of Wheat for human consumption is
given in the ISO Specification for Wheat (ISO, 2000).

STANDARDS
Standards for the control of PAs concentrate on the control of the toxic plants and plant parts.
This is especially true of plants and parts of plants used for herbal preparations and medicines,
excluded from the present discussion.

For grains and pulses, the Codex Alimentarius states that toxic seeds in wheat should not be
present in amounts that represent a hazard to health and mentions specifically the presence of
Crotolaria. These standards are:

• Maize (corn) CODEX STAN 153-1985

• Certain pulses CODEX STAN 171-1989
• Sorghum grains CODEX STAN 172-1989
• Wheat and durum wheat CODEX STAN 199-1995
• Oats CODEX STAN 201-1995

The ISO Specification for Wheat allows a tolerance of 0.05% and provides a test method (ISO,
2000). This tolerance appears to be close to the limit of determination of the method.

It appears that controls at the level of the grain are considered to the main control point and
are sufficiently effective that maximum limits for the presence of PAs in flour and derived wheat
products are considered unnecessary.

The current EU maximum levels for PAs in animal feed materials relate to weed seeds and
unground and uncrushed fruits containing alkaloids, glucosides and other toxic substances. The
maximum limits are: 3 000 mg/kg total, including 1 000 mg/kg of either or both Lolium
temulentum L., and Lolium remotum Schrank, and 100 mg/kg of Crotolaria spp (0.3%, 0.1% and
0.01% respectively based on a moisture content of 12% in the feeding stuff). It should be noted
that these data refer to seeds that can be detected by microscopic examination, but neither
differentiate between individual plants, with the exception of the three mentioned species, nor
provide limits for the amount of individual or groups of PAs.2 (European Food Safety Authority,
2007).

In the United States, animals presented for slaughter that show signs of PA-related disease, are
condemned and not allowed to enter the food supply (USDA, 2007).

CONTROL MEASURES
Measures for the control of PA-containing herbal preparations and medicines are outside the
scope of this note. Measures for the control of PA contamination of food and animal feed focus
on agricultural measures during primary production.

2 The EFSA report states that the reason for the inclusion of limits for Lolium remains unclear. Lolium,
belonging to the Graminae family, contains 2 PAs (loline and norloline); however these PAs are not known
to be hepatotoxic (International Programme on Chemical Safety, 1988).

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FAO Consumer Protection Fact Sheets
No.2: Pyrrolizidine Alkaloids

The principal control measure is weed control in accordance with Good Agricultural Practices.
According to FAO (FAO, 2007), methods most used for weed control include:

• Preventative methods (legal and quarantine procedures, and others at the farm level);
• Cultural methods (crop rotation, land preparation, use of cover crops, polycropping,
mulching, water management, hand or mechanical weeding during the crop’s life cycle);
• Chemical methods (use of herbicides);
• Biological methods (classical methods through the introduction of exotic natural
enemies and increasing the population of already existing natural enemies);
• Other non-conventional methods (soil solarization, use of hot water, and others in
development).

In cereal crops, weed control may be by the use of herbicides (e.g., 2,4 D) or by mechanical or
manual weeding of the fields. Wheat fields, millet fields, etc., should be weeded prior to planting
and periodically during the first six weeks of the growth cycle. A final weeding about two weeks
before harvest significantly reduces to possibility of contamination of the harvest with toxic
seeds. In legume crops, mechanical or manual weeding may be the only option.

The provisions in the Codex Standards for cereals and pulses for the presence of toxic seeds
should be applied BEFORE the crop is milled or distributed for human consumption. The test
method contained in the ISO standard is easily applied and does not require elaborate
laboratory equipment or extensive training of operators. It can also be applied to the Codex
Standard, bearing in mind the nature of the reporting.

In some cases, sieving can be used to separate the PA-bearing weed seeds on the basis of size.

Control of weeds in pastures, for the protection of grazing animals, should be undertaken using
the measures outlined by FAO. In the case of depletion of pastures during drought, alternative
animal feeding measures should be implemented by national and local authorities to the extent
of their capabilities.

In weeding, attention should be paid to areas bordering the crop or pasture, as these may
constitute a reservoir for the weeds and create year-after-year problems. Long-term measures
may include biological pest control but this requires extensive research and evaluation of the
environmental impact of the introduced species (North West Weeds, 2007).

Crotolaria and other PA-bearing plants are sometimes used as ground cover, soil improvers
(Crotolaria spp. are legumes) and as animal feed. The safe use of these plants as animal feed
depends on the fact that the leaves and stalks of the plant contain less PAs than the flowering
parts and the seeds. This should be taken into account when allowing animals to graze on these
plants. Also, the relative susceptibility of animals to PA intoxication (cattle being less sensitive
that sheep, for example) should be taken into account.

Finally, one of the most significant control measures is awareness and the spread of knowledge
about PAs and the implications of PA-bearing weeds and seeds in products destined for human
consumption or as animal feeds. Rural radio programmes, extension services and farmers’
associations should be considered as means of minimising the presence of PAs in foods and
feeds.

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BIBLIOGRAPHY
Anonymous. 2001. Drought causes re-emergence of liver disease. The Lancet. 2001, Vol. 358, p.
1070.

Australia New Zealand Food Authority. 2001. Pyrrolizidine Alkaloids in Food - A

Toxicological Review and Risk Assessment. Technical Report Series No.2. Canberra and
Wellington : http://www.foodstandards.gov.au/_srcfiles/TR2.pdf, November 2001.

European Food Safety Authority. 2007. Opininion of the Scientific Panel on Contamiants in
the Food Chain on a request from the European Commission related to Pyrrolizidine Alkaloids
as undesirable substances in Animal Feeds. The EFSA Journal (2007) 447, 1-51. Parma :
http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178621166892.htm, January
25, 2007.

FAO. 2007. Reccomendations for Improved Weed Management. Rome : Food and Agriculture
Organization of the United Nations, ftp://ftp.fao.org/docrep/fao/010/a0884e/a0884e00.pdf,
2007.

International Programme on Chemical Safety. 1988. Pyrollizidine Alkaloids. Environmental

Health Criteria NO.80 (EHC 80). Geneva : WHO.
http://www.inchem.org/documents/ehc/ehc/ehc080.htm, 1988.

ISO. 2000. Wheat (Triticum aestivum L.) ISO 7970:2000. Geneva : International Organization for
Standardization, 2000.

K.A. Than, V. Stevens, A. Knill, P.F. Gallagher, K.L. Gaul, J.A. Edgar, S.M. Colegate. 2005.
Plant-associated toxins in animal feed: Screening and confirmation assay development. Animal
Feed Science and Technology. 2005, Vol. 121, pp. 5-21.

K.L. Stuart , G. Bras. 1957. Veno-occlusive disease of the liver. Cited in "Pyrrolizidine Alkaloids
in Foods" (ANZFA, 2001). Canberra and Wellington : Australia New Zealand Food Authority,
1957.

North West Weeds. 2007. Blue Heliotrope. North West Weeds. [Online] November 25, 2007.
[Cited: April 15, 2008.] Government of New South Wales
http://www.northwestweeds.nsw.gov.au/blue_heliotrope.htm.

USDA. 2007. Multi-species Disposition Basics with a Public Health Focus. s.l. : United States
Department of Agriculture, Food Safety Inspection Service
http://www.fsis.usda.gov/PDF/PHVt-Multi_Species_Disposition.pdf, 2007.

WHO. 2001. Health Talks Afghanistan. Geneva : World Health Organization, September 2001.

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