U. S.

ENVIRONMENTAL PROTECTION AGENCY

ISSUE PAPER

NATURAL TOXICANTS IN FOOD FROM PLANTS

Revised December 2019

Less than 1/10th of 1% of all the constituents of food have been determined to have the potential
at higher levels of exposure to cause adverse effects in humans. In this report, the Environmental
Protection Agency (EPA) evaluates those natural toxicants, approximately 209 at last estimation,
originally identified in a report from the International Food Biotechnology Council (IFBC)1.
Most varieties of the food plants referred to in this paper used in the United States have been
bred to reduce toxicant concentration levels below those known to cause adverse effects.

Of the natural toxicants identified in the IFBC report, only 10% (21) have been shown to be in
foods consumed as part of a normal diet and to cause harm when present in high concentrations.
The normal diet does not include plants consumed during times of deprivation, as part of a
religious diet, or through misidentification or substance abuse. Approximately 50% of those
present in the normal diet (11) are “pass-through” contaminants in honey or milk; produced from
pollen of the rhododendron, azalea, yellow jasmine of the tutu tree, or from cows grazed on
hemlock species, respectively1. None of these plants are used or are intended to be used as food
plants. Additionally, one toxicant (nitrite) can become concentrated in plant leaves after uptake
from nitrate-rich fertilizer. This toxicant is not considered a biosynthesized toxicant and falls
outside of the potential exemption.

The remaining toxicants (9) with the potential to cause adverse effects when consumed at high
concentrations are discussed below. This paper also addresses toxicants identified in comments
(9) received2 in response to the 1994 proposal (59 FR 60495) on the proposed exemption of
plant-incorporated protectants moved amongst plants in sexually compatible populations: Two of
the toxicants identified in the 1995 comments were among the 21 originally identified in the
IFBC report.

Amino Acid Analogues

Hypoglycin A found in immature ackee fruit.

β-N-oxalylamino-L-alanine found in chickling vetch, vetchling.

1
Coulson, F. and A. C. Kolbye, Jr. (eds) Regulatory Toxicology and Pharmacology, 12(3), S20-S30. 1990.
2
Hanson, M and J. Halloran. Letter on dockets OPP-300367a through OPP300371a. February 22, 1995.
General: Hypoglycin A is an unusual, toxic amino acid which inhibits fatty acid metabolism.
Hypoglycin A acts as an inhibitor of acyl-CoA dehydrogenase, restricting the available amount
of Coenzyme A, and eventually inhibiting gluconeogenesis. This can result in severe
hypoglycemia and vomiting. While the immature akee fruit contains high levels of this toxin
(>1000 ppm), these levels are reduced to undetectable ranges upon fruit ripening. This fruit is
primarily consumed in Africa and Jamaica3. Hypoglycin A is heat stable, and is not reduced by
cooking. The best method to prevent adverse effects is avoidance of unripe fruit.

β-N-oxalylamino-L-alanine (BOAA) is a glutamate analog. Current evidence suggests that free

radicals release by this excitatory neurotoxin interact with neuronal glutamate (non-NMDA
subtype) receptors, resulting in spastic paralysis. There is some evidence that free radical
scavengers (e.g., vitamin C) may have a protective effect4. Chickling vetch is primarily
consumed on the Indian sub-continent. Vetchling is grown primarily in Spain, but human
consumption is prohibited by the Spanish Food Code. This toxin can mostly be leached out of
seeds by soaking in water, followed by cooking. Chickling vetch and vetchling are also grown
for feed, although they are not extensively used for fear of poisoning5.

Regulatory Status: FDA issued a worldwide import alert in 1973 which banned the akee fruit
from entering the United States. An import alert was issued by the FDA on 5/31/95. While the
notice did not establish any requirements, all akee fruit products were to be automatically
detained due to the potential for the presence of high levels of hypoglycin A and B in the fruit,
and because of the difficulties in separating immature from mature fruit in processed foods. This
alert was updated on 7/3/00 to include the detainment without physical examination of akees in
all product types due to contamination by natural toxins except for canned akee originating from
producers listed in an attachment to the alert determined to have food safety procedures in place.
The most recent import alert on akee fruit was issued on 06/03/106; it also listed the producers
determined to have appropriate safety procedures in place.

No information on the regulatory status of β-N-oxalylamino-L-alanine in the United States could

be found.

Current Testing Methods: An ion-exchange chromatographic method with the capacity to

separate hypoglycin A from other amino acids was developed by the FDA in 1989 to detect this
toxicant in canned akee fruit7. The Agricultural Research Services of the USDA developed
monitoring strategy for hypoglycin A in 20078.

3
Peter R. Cheeke. Natural Toxicants in Feeds, Forages, and Poisonous Plants. Interstate Publishers, Danville, IL.
1998. p. 432-433.
4
Dunham, W., Tsao, C., Barth, R., and Z. Herman. Protection by Dietary Ascorbate of Guinea Pigs from
Neurolathyrism. Nut. Res. 15:993-1004. 1995.
5
J. E. Hernandez Bermejo and J. Leon (eds) Neglected Crops: 1492 from a Different Perspective. FAO Plant
Production and Protection Series No. 26. Rome, Italy, p 273-288. 1994.
6
http://www.accessdata.fda.gov/cms_ia/importalert_64.html
7
Chase, G. W., Landen, W.O. Jr., Gelbaum L.T., and A.G. Soliman. Ion-exchange Chromatographic
Determination of Hypoglycin A in Canned Akee Fruit. J. Assoc. Anal. Chem. 72(2):374-7. 1989.
8
Whitaker, T., Saltsman, J., Ware, G., and A. Slate. J AOAC Int. 90:4 2007.

2
Methods for measuring BOAA include high performance liquid chromatography7, as well as
positive chemical-ionization gas chromatography-mass spectrometry9.

Glycosides

Vicine, covicine found in fava beans.

General: The active metabolites are thought to be the aglycone (sugarless) derivatives of vicine
and covicine, divicine and isouramil. The effects of vicine and covicine are mediated by
decreasing the levels of the cellular reducing agent, glutathione. Glutathione is important for
reducing a by-product of cellular reactions, the strong oxidant hydrogen peroxide. Lowered
levels of glutathione result in a build-up of hydrogen peroxide, eventually leading to degradation
of red blood cells and hemolysis. Individuals with the inherited Mediterranean form of G6PD
(glucose-6-phosphate dehydrogenase) deficiency, which also results in lowered levels of cellular
glutathione, are particularly sensitive to foods containing vicine and covicine. The combined
stresses on glutathione levels in this sensitive subpopulation can result in a form of hemolytic
anemia. Hemolytic episodes are averted by warning from physicians and by avoidance of these
foods. Occurring episodes can be managed10. Toxicant levels can be reduced by soaking, as well
as by removal of the fava bean skin11.

Regulatory Status: No information on regulatory status in the United States could be found.

Current Testing Methods: Levels of vicine and covicine can be measure by high performance
liquid chromatogrphy12. Alternatively, the glucosides can be extracted and hydrolyzed by beta-
glucosidase. The aglycone (sugarless) products are then used to reduce a compound, o-
ferriphenanthroline, in a reaction which can be followed spectrophotometrically at 515nm13.

Steroid Terpenes

Cucurbitacin found in cucumbers, yellow squash, watermelon.

General: As many as 17 different members of the cucurbitacin class of natural toxicants can be
found as naturally occurring mixtures in members of the Cucurbitaceae family. Cucurbitacin can
result in a bitter taste and may cause stomachaches or cramps at extremely high concentrations1.
Most cultivated varieties have extremely low concentrations of cucurbitacin, although occasional
pollination from wild varieties can occur, resulting in slightly higher concentrations of toxicant

9
Pan, M., Mabry, T.J., Cao, P., and M. Moini. Identification of nonprotein amino acids from cycad seed as N-
ethoxycarbonyl ethyl ester derivatives by positive chemical-ionization gas chromatography- mass spectrometry. J.
Chromatogr A. 787:288-294. 1997.
10
Cooper, R. A. and H.F. Bunn. Hemolytic Anemias. In: Harrison’s Principles of Internal Medicine. Eds..
Isselbacher, K.J., R.D., Adams, E. Brannwald, R.G. Petersdorf and J.D. Wilson. McGraw-Hill Company. 1980.
11
Abd Allah, M.D., Foda, M.H., Abu Salem, F.M., and Z.S. abd Allah. Treatments for Reducing Total Vicine in
Egyptian Faba Bean (Giza 2 Variety). Plant Foods Hum Nutr. 38:201-210.1988.
12
Lattanzio, V., Bianco, B.B., and D. Lafiandra, D. High-Performance Reversed Phase Liquid Chromatography
(HPLC) of Favism-Inducing Factors in Vicia faba L. Experientia. 38:789-790. 1982.
13
Chevion, M. and T. Navok. A Novel Method for Quantitation of Favism-Inducing Agents in Legumes. Anal
Biochem. 128:152-158. 1983.

3
in produce. Cucurbitacin acts as a feeding stimulant in the cucumber beetle14 and is used as a
flavor additive in pesticides directed towards this pest.

Regulatory Status: Residues of cucurbitacin from the root powder of the buffalo gourd (57 FR
40128), the juice of zucchini (63 FR 43085), or the powder or juice of the Hawkesbury melon
(65 FR 15248) are exempt from the requirement of a tolerance (40 CFR 180) on food crops when
used as an inert (gustatory stimulant). Current limits for application are 3.4 g
cucurbitacin/acre/season.

Current Testing Methods: EPA concluded that an analytical method was not required for
enforcement of cucurbitacin from Hawkesbury melon (65 FR 15248). However, the Agricultural
Research Service of the USDA has developed a high performance liquid chromatography
(HPLC) method for analyzing the concentration of cucurbitacin E in juice or in reconstituted
residues from the Hawkesbury melon15.

Steroid Glycoalkaloids

Solanine, chaconine found in potatoes, eggplant, Jerusalem cherries and to a much lower
extent in peppers, apples and sugar beets.
Tomatine found in green tomatoes.

General: Solanine and chaconine are members of a large class of naturally occurring
glycoalkaloids found in the nightshade family, and impart a bitter taste to food. Gastrointestinal
and neurological symptoms of glycoalkaloid poisoning derive from the two primary toxic effects
associated with the glycoalkaoids; membrane disruption and acetylcholinesterase inhibition.
When tested individually, members of this class display different activities with respect to the
two identified toxic effects. In vitro experiments indicate that the glycoalkaloid solanine displays
weak to moderate cholinesterase inhibition as well as membrane disruption, while the
glycoalkaloid chaconine acts primarily as a membrane disruptor. Exposure to the glycoalkaloids
is, however, invariably to mixtures of the glycoalkaloids normally found in produce from
members of the Solanaceae family. The term “solanine” is generally used to represent
aggregated glycoalkaloids.

While glycoalkaloid poisoning is rare16, large oral doses of glycoalkaloid mixtures can cause GI
tract irritation including moderate nausea, vomiting, diarrhea, internal hemorrhaging, and edema.
These effects usually occur within 8-12 hours after ingestion17. In vivo testing with animals

14
US EPA Issue Paper. FIFRA: Benefit and Environmental Risk Considerations for Inherent Plant Pesticides. 1994.
15
Matsuo, K. Demilo, A.B., Schroder, R.F.W., and P.A.W.Martin. Rapid High-Performance Liquid
Chromatography Method to Quantitate Elaterinide in Juice and Reconstituted Residues from a Bitter Mutant of a
Hawkesbury Watermelon. J. Ag. and Food Chem. 47:2755-2759. 1999.
16
Smith, D.B., Roddick, J.G. and J.Leighton Jones. Potato Glycoalkaloids: Some Unanswered Questions. Trends in
Food Science and Technology. 7, 126-131. 1996.
17
R. Tice. α-Chaconine [CAS # 20562-03-2] and α-Solanine [CAS # 20562-02-1]; Review of Toxicological
Literature. Prepared for E. Zeiger, National Toxicology Program, National Institute for Environmental Health
Sciences, National Institutes of Health. 1998. [http://ntp-
server.niehs.nih.gov/htd...und/ExecSumm/ChaconineSolanine.html] R. Tice. α-Chaconine [CAS # 20562-03-2]
and α-Solanine [CAS # 20562-02-1]; Review of Toxicological Literature. Prepared for E. Zeiger, National

4
shows the primary toxic effect upon ingestion of glycoalkaoid mixtures to be membrane
disruption18. These natural toxins are rapidly excreted from the body in urine and feces, but can
accumulate to a small degree in abdominal fat, adrenals, blood, brain, heart, kidney, liver, lungs,
muscles, pancreas, spleen, testes, thymus and thyroid. Intraperitoneal or intravenous injection of
glycoalkaloids (primarily solanine) into test animals also produces nervous system impairment;
including drowsiness, changes in blood pressure and heart rate. The LD50 of an oral dose is
approximately 10 times larger than that of an intraperitoneal dose, which is larger than the LD50
of an intravenous dose18.

Solanines (e.g., aggregated glycoalkaloids) are concentrated in portions of the plant undergoing
active metabolism; the leaves, sprouts, any bruised/wounded portions of the tuber and in the
skin. In an undamaged, unsprouted tuber, 30-80% of the solanines are found in, and directly
under the skin19. Peeling, or removing the damaged portion of the potato, is the best way to
reduce solanine levels. Cooking will not effectively remove all of the solanines present. There
is disagreement in the literature as to the efficacy of boiling in steam or water to remove
solanines. While some reports indicate this treatment has no effect on alkaloid levels, others
indicate that this treatment can reduce concentrations by 30-40 %20. Deep frying in oil at
170oC17 and irradiation21 have been shown to lower solanine concentrations.

At concentrations of 10 mg/100 g potato, solanines impart a bitter taste. At concentrations above

20 mg/100 g potato, solanines can leave a persistent irritation and burning sensation on the
tongue22. The lethal dose is estimated at 3-6 mg/kg body mass17,23,24. Of 32 potato varieties on
the U.S. market studied, the average concentration of solanines was 7.5 mg alkaloid/100 g tuber.
The average consumption of solanines is estimated at approximately 12.5 mg
alkaloid/person/day22. To consume the minimum lethal dose, assuming 100 % of the
glycoalkaloid present in a potato tuber (medium potato of 148 gm) would be absorbed upon
ingestion, which is unlikely as solanine, chaconine and their common derivative solanidine are
absorbed extremely poorly from the GI tract25, a person weighing 150 lbs (~68 kg) would have
to consume approximately 6 lbs of potatoes (2,723 g, approximately 18.4 potatoes) in one
sitting.

Toxicology Program, National Institute for Environmental Health Sciences, National Institutes of Health. 1998.
[http://ntp-server.niehs.nih.gov/htd...und/ExecSumm/ChaconineSolanine.html]
18
U.S. EPA Issue Paper. The Glycoalkaloid Class; Solanine and Chaconine: Mechanisms of Action. 2000.
19
Bushway, R.J., Bureau J.L., and D.F, McGann. Alpha-chaconine and Alpha-solanine Content of Potato Peels and
Potato Peel Products. J Food Sc. 48(1): 84-86.
20
Wayne Vandre. Greening of Potatoes, FGV-00337. Alaska Cooperative Extension.1995.
[http://www.uaf.edu/coop-ext/publications/freepubs/FGV-00337.html]
21
R.Y. Yada and A.G. Marangoni. AG2056 - Improving the Organoleptic Quality of Irradiated Food. Reasearch
Projects Summary. Ministry of Agriculture, Food and Rural Affairs. Ontario, Canada. 1999.
[http://www.gov.on.ca:80/OMAFRA/engl.../researchfund/agfooddocs/ag2056.htm]
22
R. C. Beier. Natural Pesticides and Bioactive Components in Foods. Rev Environ Contam Toxicol. 113, 75-84.
1990.
23
R. Houben and K. Brunt. RP-HPLC Analysis of Glycoalkaloids in Potato Tubers. Proceedings from Starch ‘96,
Leeuwenhorst, Noordwijkerhout, The Netherlands. 1996.
24
Morris, S.C. and T.H. Lee. The Toxicity and Teratogenicity of Solanaceae glycoalkaloids, particularly those of
the Potato (Solanum tuberosum):a Review. Food Tech Australia. 36:118-124. 1984
25
Groen, K., Pereboom-De Fauw, D.P.K.H., Besamusca, P., Beekhof, P.K., Speijers, G.J.A., and H.J.G.M. Derks.
Bioavailability and Disposition of 3H-Solanine in Rat and Hamster. Xenobiotica 23:995-1005. 1993 show the
bioavailability of the glycoalkaloid solanine is 1.6% in the rat, and 3.2 % in the hamster.

5
Similarly, a child weighing 50 lbs (22.5 kg) would have to consume 2 lbs of potato (908 g, or
approximately 6.1 potatoes) in one sitting.

Regulatory Status: Currently there are no regulations in the United States mandating solanine
levels present in food derived from members of the Solanaceae family. However, the accepted
guideline established by the USDA is a maximum concentration of 20 mg/100 g of potato (cited
in Beier, 1990).

Section V(A) of the FDA “Statement of Policy: Foods Derived from New Plant Varieties” (57
FR 22984) explains that new plant varieties are not routinely subjected by FDA to scientific
tests, and that established practices by breeders have historically proven sufficient to ensure food
safety. If those practices cannot assure that toxicant levels have not been elevated above those
deemed as “safe”, analytical tests may be appropriate. Under Title 21 of the Code of Federal
Regulations (Food and Drugs), it is the responsibility of the producer to ensure that the food they
offer is safe and meets the requirements of the law.”

In accordance with the above statements, most potato varieties displaying unacceptable levels of
glycoalkaloid are rejected during breeding efforts, and not developed into marketable products.
In order to reduce the number of potatoes released to the market which may have unacceptable
levels of glycoalkaloids due to environmental conditions, common screening procedures are
conducted during the grading and shipping of potatoes. USDA grade standards are voluntary,
although some states have regulations requiring grading. Potatoes with a green coloration are
considered to have a higher solanine content. (The green color is derived from chlorophyll and
not from solanine, but indicates the presence of ongoing metabolism, and the production of
solanines, in the tuber. Greening occurs upon exposure to light through either inadequate soil
coverage during growth or during storage.) USDA potato grades consider a potato damaged
when removal of the green portion would result in a loss of 5% if the total potato weight.
Potatoes are considered seriously damaged when removal of the green portion would result in a
loss of 10% of the total potato weight.

Current Testing Methods: Currently most chemical testing is accomplished by broad spectrum
analysis through gas or reverse-phase high performance liquid chromatography26. A new
monoclonal antibody system, for which patent protection is currently pending, has been
developed by the USDA. This assay system may provide quicker and more convenient testing.
The antibody based kit would allow testing of raw or processed potato products, as well as high
throughput screening by breeders, producers and processors27.

Cyanogenic Glycosides

Linamarin found in cassava.

Phaseolunatin found in lima beans.
Amygdalin, prunasin found in almonds, apples, apricot, cherry, peach pear plum.
Dhurrin found in sorghum.

26
R. Houben and K. Brunt. RP-HPLC Analysis of Glycoalkaloids in Potato Tubers. Proceedings from Starch ‘96,
Leeuwenhorst, Noordwijkerhout, The Netherlands. 1996.
27
Marcia Wood. New Safeguards Against Glycoalkaloids. Agricultural Research Magazine 45 (12), 16-17, 1997.

6
General: Cyanogenic glycosides are enzymatically processed either within the human body, or
upon wounding (crushing) of the plant part, to hydrocyanic acid (HCN). Cyanide is a metabolic
poison, inhibiting the action of cytochrome-oxidase, preventing the utilization of oxygen in
cellular respiration and ultimately oxidative phosphorylation. Poisoning by cyanide can result in
neurological disorders, breathing difficulties, thyroid enlargement and, in extreme cases, coma
and death28. The body can accommodate quantities of cyanide found in normal dietary foods by
metabolizing it to thiocyanate or by combining it with cysteine29.

Traditional methods of processing cassava to reduce the cyanogenic effects include peeling,
washing/soaking, boiling, drying and fermentation30. Lima beans are rendered non-toxic by
cooking31. Amygdalin and prunasin are generally found in portions of the plant not normally
consumed as food (the pit/seed/kernel as appropriate), except for almonds which are regulated as
indicated below. Sorghum is generally used for feed. Silage made from this plant is generally
safe, as the sorghum is dried in processing and the HCN is volatile32.

Regulatory Status: No information on the regulatory status of cyanogenic glycosides in the

United States could be found.

However, hydrogen cyanide is regulated as a food additive (40 CFR 185.3600) at 200 ppm in
cocoa (from use as a fumigant), 125 ppm in cereal flours, 90 ppm in cereals cooked before
consumption and 50 ppm in uncooked ham, bacon and sausage33.

Bitter almonds, due to their potential to cause cyanide poisoning, may not be marketed in the
United States. Sweet almonds cannot contain more than 5% bitter almonds. Almond paste (or
other pastes containing almond paste) must have less than 25 ppm HCN34.

While no established standards for phaseolutinan levels could be found, lima beans in the United
States were limited to products producing less than 10-20 mg HCN/100 g35. Lima beans
currently grown in the United States are within this range36. Section 8e of the Agricultural

28
R. Eisler. Cyanide Hazards to Fish, Wildlife and Invertabrates: A Synoptic Review. Biological Reports 85,
Containment Hazard Review Reports. U.S. Department of Interior, Fish and Wildlife Service. Washington D.C.
1991. p.1-9.
29
R. Hall. Toxicants Occurring Naturally in Foods. NAS, Washington D.C. p. 164-173. 1966.
30
M.R. Grace. Cassava Processing. FAO Plant Production and Protection Series No. 3. Sections 1, 5.4, 6 and
7.1998.
31
I. Liener. Toxicants Occurring Naturally in Foods. NAS, Washington D.C. p. 58-61. 1966.
32
Peter Cheeke and Lee R. Shull. Natural Toxicants in Feeds and Poisonous Plants. AVI Publishing Company, Inc.,
Westport,CT. 1985. a) p.175. b) p. 186-189.
33
Food Additive Status List. FDA. [http://www.fda.gov/ora/inspect_ref/iom/exhibits/ApAMS9.html
34
Requirements of Laws and Regulations Enforced by the U.S.Food and Drug Administration. (Summary and
cross-reference to the major requirements enforced by FDA.), Section I: Foods; Nuts and Nut Products.
[http://www.fda.gov/opacom/morechoices/smallbusiness/blubook.htm#toc.html]
35
cited in: Montgomery, R.D. Observations on the Cyanide Content and Toxicity of Tropical Pulses. West Indian
Med. J. 13 (1) pg 1. 1964.
36
Jose M. Concon. Food Toxicology: Principles and Concepts. Marcel Dekker, Inc. New York and Basil. 1988. p.
283.

7
Marketing Agreement Act of 1937 indicates that regulations applied to domestic foods also
apply to imported foods.

Current Testing Methods: Currently, both a qualitative (colorimetric) assay and a quantitative
(titration) testing method exist for hydrocyanic (prussic) acid. Quignard’s test (qualitative
method) relies on the rapidity of development of an orange color on a piece of sodium picrate
paper to indicate the amount of hydrocyanic acid present. More quantitative measurements of
hydrocyanic acid levels present in fresh tuber can be obtained by extraction, distillation, and
alkaline titration with silver nitrate. (1 mL of silver nitrate utilized is approximately equal to 1
mg of hydrocyanic acid)30.

Official methods of analysis for cyanide in distilled liquors, for HCN in almond extract or for
cyanogenic glycosides in feeds (all quantitative, titration) are detailed in the Official Methods
Analysis of the Association of Official Analytical Chemists37a,b,c.

Quinolizidine Alkaloids

Sparteine, lupinine, lupanidine found in lupines, flax.

General: Sparteine blocks nicotinic cholinergic receptors and is a weak muscarinic cholinergic
receptor antagonist38. Plants containing sparteine (e.g., Scotch Broom) have been used in herbal
medicine apparently as a sodium channel blocker. Lupines are a traditional grain legume utilized
by indigenous cultures in South America and the Mediterranean; in these areas today cultivated
varieties are grown for consumption which have low levels of sparteine in the grain. Lupines are
utilized primarily as forage or feed for grazing animals in the United States. Poisoning (of
livestock) through consumption of wild species of lupine is associated with breathing problems,
weakness, and loss of muscular control39. Furthermore, ingestion of lupines within the first
trimester of pregnancy has teratogenic effects, resulting in “crooked calf” syndrome (skeletal
defects) and craniofacial malformations40. A few cases have been reported where the ingestion
of milk, from cows and/or goats fed lupines, by humans or animals during the first trimester of
pregnancy has also led to similar malformations40. Alkaloids in these types of exposures may be
classified as “pass-through” contaminates. The alkaloid responsible for these teratogenic effects
is found only in wild lupine species3. Treatment for accidental consumption is supportive.
Traditional processes for removal of these alkaloids from legume grain include soaking and
cooking. Newly bred variants of sweet lupines are grown extensively in Germany; they lack any
bitter taste and require no soaking in salt solution prior to ingestion. The sweet varieties of
lupine contain <200 mg alkaloid/kg compared to bitter varieties that contain 5-40 gms

37
P. Cunniff ed. Official Methods of Analysis of AOAC (Association of Official Analytical Chemists) International.
16th ed.1995. a)cyanide: Chap. 26, p. 17-18. b)HCN: Chap. 36, p. 18, Chap. 49, p. 49. c)cyanogenic glycosides: Chap.
49, p. 49. d)coumarin: Chap. 28, p.15-16, Chap. 36, p. 4-5. e)tannins: Chap. 26, p. 16-17.
38
D. G. Barceloux. MD Medical Toxicology of Natural Substances. 784-787. 2008 John Wiley and Sons.
39
Bulletin 415:Poisonous Plants of the Western United States; Neurotoxic and Mycotoxic Plants. USDA/ARS
Poisonous Plant Research Lab. [http://www.pprl.usu.edu/bulletin.htm]
40
Ames, B., Profet, M. and L.S. Gold Nature's Chemicals and Synthetic Chemicals; Comparative Toxicology.
PNAS 87, 7782-7786, 1990.

8
alkaloid/kg38. The United States has a developing specialty human food market for lupine in the
form of lupine flour, lupine pasta, and hulls for dietary fiber.

Regulatory Status: No information on regulatory status in the United States could be found.
However, the lupines are cultivated in Australia, South America, Eastern Europe and the
Mediterranean region with regulatory limitations of 100 μg alkaloid/gm material.

Current Testing Methods: Analytical methods for the detection and quantitation of lupine
alkaloids include gas chromatography with flame ionization or nitrogen phosphorus detection or
gas chromatography/mass spectrometry. A high performance liquid chromatographic method is
more sensitive and can detect lesser amounts38.

Phenolic Compounds

Gossypol found in cottonseed.

Myristicin found in nutmeg (mace), dill, parsley, celery, carrots.
Coumarin found primarily in Tonka beans, also in lavender and woodruff.
Psoralens (furocoumarins) found in celery, fennel, parsnip, coriander, parsley, dill, citrus fruit
(lime, lemon, orange, grapefruit).
Tannins found in a wide range of foods including fruits (pigment), beans (kidney, black),
coffee, tea, sorghum.

General: Gossypol acts as an inhibitor of several dehydrogenase enzymes. It has pro-apoptopic

qualities probably through the inhibition of some of these enzymes. Cottonseed and cottonseed
meal are important sources of protein utilized for feed in ruminants, which appear to be less
sensitive to gossypol concentrations than non-ruminants as ruminant microflora can digest
gossypol to some extent. Depression, loss of weight, diarrhea, cardiac irregularity, and various
blood disorders (e.g., hemolytic anemia) have been associated with gossypol poisoning of
livestock. Cottonseed can be detoxified to varying degrees by cooking, steaming, autoclaving,
and through utilization of other chemicals means (e.g. treatment with alkali)41. Researchers at
Texas A&M have genetically engineered cotton plants that contain very little gossypol in the
seeds, but retain higher levels in the leaves and stems42.

Myristicin is primarily found in the oil produced from the plant Myristica fragans. Nutmeg is
the dried ripe seed while mace is the dried aril which encloses the shell containing the seed.
Myristicin has been indicated as a weak monoamine oxidase (MAO) inhibitor. Excessive use of
nutmeg has caused clinical intoxication, characterized by nausea, vomiting dizziness, anxiety,
headache, hallucinations and irrational behavior. High dosage (1 -3 whole nuts, or 5-15g grated
spice) of this compound can result in hallucinations, headaches, cramps, nausea, liver damage,
and even death. The LD50 for oral dosage in rats is 4260mg/kg. Toxic effects usually occur in
3-6 hours following ingestion of high doses. Most recovery is usually within 24 hours43. This

41
E. Eagle. Toxicants Occurring Naturally in Foods. NAS, Washington D.C. p. 242-249. 1966.
42
Cotton Seed Protein: From Farmers to your Family Table. November 22, 2006. MedGadget.
[http://www.medgadget.com/archives/2006/11/cottonseed_prot.html]
43
CAS Number 607-91-0. NTP Chemical Repository. 1997. [http://ntp-server.niehs.nih.gov/].

9
dosage is, however, much greater than any used for flavoring. Poisoning by spice/flavor is
therefore extremely rare29. Also, while myristicin has been widely accepted as the main
psychoactive component of nutmeg, both the differences in subjective effects observed between
nutmeg and synthetic myristicin, as well as the fact that myristicin is not the main component of
the seed (therefore is possibly not present in high enough quantities) suggest that it alone does
not fully explain the effects of consuming raw nutmeg44.

Coumarin may be found in many plants, notably in high concentration in the tonka bean, vanilla
grass, woodruff, mullein and sweet grass. It has a sweet scent, readily recognized as the odor of
newly mowed hay. Although it has a sweet fragrance, it has a bitter taste. Coumarin may be
found in substances used as flavoring (e.g. artificial vanilla extract from Mexico). Coumarin is
used as a fixative or enhancing agent for aroma, and is therefore found in small amounts in
perfumes, soaps, toothpastes and hair preparations. It is also used to fix the flavor and aroma in
a number of tobacco products45. However, it is not used as a food additive in the United States
after findings of extensive liver damage in rats fed high dosages (see Regulatory Status
section)29. Although coumarin is a potent rodenticide, it is only moderately toxic to humans.
Rats and other rodents metabolize it to 3,4-coumarin epoxide while humans metabolize it to 7-
hydroxycoumarin, a compound of lower toxicity. Coumarin acts by blocking the production of
Vitamin K, eventually resulting in a decreased activation of thrombin and clotting in response to
wounding. Symptoms of coumarin poisoning range from extended blood-clotting time to
hemorrhaging and death32b. The oral LD50 in rats is 293 mg/kg46.

Psoralens are contained in many plants and are precursors to furocoumarins (coumarin with a
furan ring). Psoralens can cause skin photosensitization including burning sensations, dermatitis,
and hyperpigmentation. Psoralens (supervised and in small doses) are commonly utilized in
treatment for a variety of skin diseases, including vitiligo and psoriasis46. Some psoralens (e.g.,
methoxypsoralen used in PUVA treatment of psoriasis and vitiligo) are identified as carcinogens,
resulting in increased incidences of various skin cancers, including basal and squamous cell
carcinomas . Due to their relative hydrophobicity, psoralens applied topically can penetrate into
skin cells, where they intercalate into the DNA. Subsequent exposure to sunlight (UV light)
causes the DNA pyrimidine bases to crosslink, preventing DNA synthesis and resulting in skin
blisters and rashes (apoptosis) and potentially producing DNA mutations. The average daily
dietary intake of total furocoumarins is estimated at 1.3 mg/day47. While the primary exposure
route is dietary, the primary exposure route leading to adverse effect is dermal. Healthy celery
contains approximately 10-100 ug/g wet weight psoralens. 1 ug psoralen/cm2 exposed skin can
produce blisters after 10 minutes exposure to bright sunlight48. Dermal exposure through the
small amount of exudate on the cut ends of celery, or other foods containing psoralens at levels
commonly observed in varieties grown in the US, is unlikely to produce the quantities of
psoralen required to cause an adverse effect.

44
http://ntp.niehs.nih.gov/ntp/htdocs/Chem_Background/ExSumPdf/Myristicin.pdf
45
CAS Number 91-64-5. NTP Chemical Repository. 1991. [http;//ntp-
db.niehs.nih.gov/NTP_Rep...hem_H&S/NTP_Chem9/Radian91-64-5.txt].
46
P. McClelland, Morgan, P., Leach, E. E. and J. Shelk. Psoralen Photochemotherapy. Dermatol Nurs 9(6) 403-415.
1997.
47
J.D. Wagstaff. Dietary Exposure to Furocoumarins. Reg Tox and Pharm. 14:261-272. 1991.
48
Lovell, CR. Plants and the Skin. First ed. Oxford, Blackwell Scientific Publications, 1993.

10
Tannins are a diverse group of polyphenolic and flavanoid compounds that are widely dispersed
throughout the plant kingdom. As such, they are part of the human and animal diet. While
dietary intake has not been estimated directly, it has been suggested to be as high as hundreds of
mgs/day49. Tannins act by precipitating proteins, reducing the palatability of and nutrient uptake
from food and thereby negatively effecting growth. To compensate for tannin compounds in
tannin-rich diets, some animals produce mucin proteins for protection. [Mucin proteins
specifically bind tannins, reducing their toxicity]3. Although mostly considered to have negative
effects due to their growth inhibiting properties, tannins are increasingly being investigated for
potential health benefits as anti-oxidants and in preventing heart disease50.

Regulatory Status: It is generally accepted that cottonseed meals containing less than 0.04%
gossypol are safe for animal consumption41. In addition, a color additive order was issued
(revised 1999) allowing the use of cottonseed flour or other cottonseed products with less than
0.045% gossypol (by weight) for human consumption51.

No information on the regulatory status of myristicin in the United States could be found. No
standards or guidelines have been set by NIOSH or OSHA for occupational exposure to or
workplace allowable levels of myristicin. The American Conference of Governmental Industrial
Hygienists (ACGIH) has not recommended a threshold limit value (TLV) or biological exposure
index (BEI) for myristicin44.

Coumarin is listed by the FDA as among Substances Generally Prohibited From Direct Addition
or Use as Human Food as per 21 CFR 189.130 (b). Food containing any added coumarin as such
or as a constituent of tonka beans or tonka extract is deemed to be adulterated under the act,
based upon an order published in the Federal Register of March 5, 1954 (19 FR 1239). However,
some natural additives containing coumarin (such as sweet woodruff) are allowed in alcoholic
beverages only (21 CFR 172.510).

Psoralens are major constituents of bergamot oil (present in some perfumes, sunscreens and food
products). In the Hazardous Substances and Articles Act (16 CFR 1500.13) products containing
2% or more bergamot oil are identified as "strong sensitizers". Bergamot oil is listed by the EPA
as an approved Floral Attractant/Plant Volatile pesticide active ingredient (6/99,
http://www.epa.gov/oppbppd1/biopesticides/ai/floral_attract.htm).

The following information was found on tannic acid:

27 CFR 24.246 Subpart L Materials Authorized for Treatment of Wine and Juice
In apple juice or wine and in red wine, tannins cannot exceed 3.0 g/L gallic acid equivalents
(GAE). In white wine, tannins cannot exceed 0.8 g/L. It is considered GRAS according to FDA
advisory opinions 4/6/59 and 3/29/60. However, added tannic acid cannot increase total tannins
above the limit of 150 milligrams/liter.

49
A. King and G. Young. Characteristics and Occurrence of Phenolic Phytochemicals. J Am Diet Assoc 99(2) 213-
218. 1999.
50
L. Bravo. Polyphenols: Chemistry, Dietary Sources, Metabolism, and Nutritional Significance. Nut Rev 56(11)
317-333. 1998.
51
Code of Federal Regulations. Title 21, part 73.140(b) and 172.894(b)(1).

11
21 CFR 73.170. The section stipulates that grape skin extract (“containing common components
of grape skin juice; namely . . . tannins. . . ”) is exempt from certification as a color additive.

40 CFR 180.1001 Tolerances and Exemptions from Tolerances for Pesticides in Food Subpart d
(list of materials exempt from tolerance as inert ingredients in pesticide formulations). Tannin is
exempt under this category and is used as a dispersant.

Current Testing Methods: The following information has been found for this group of
toxicants.

Gossypol can be tested by several methods. Currently, monoclonal antibody testing is described
in the Official Methods of Analysis of the Association of Official Analytical Chemists37.

Myristicin can be measured by gas chromatography43.

Gas chromatography is also used for detecting coumarin in food (wine and vanilla), as detailed in
the Official Methods of Analysis of the Association of Official Analytical Chemists37d.

Although many methods have been established for testing tannins, most are unsatisfactory due to
the complexity of the tannin class of molecules, which can range in molecular weight from 500
to over 3000. In addition, many methods measure soluble, extractable tannins and do not
account for insoluble tannins. (Insoluble tannins may have equal biological activity as soluble
tannins.) The official method for detecting tannins in distilled liquors (spectrophotometric
analysis) is detailed in the Official Methods of Analysis of the Association of Official Analytical
Chemists37e.

Other Toxicants

The Agency finds that the other toxicants identified in the IFBC report as having the potential to
adversely affect human health not discussed here lie outside the scope of this paper pertaining to
food-use plant toxicants. These toxicants are not considered to be part of the normal diet, or are
not directly biosynthesized by the plant. Natural toxicants are well recognized by millennia of
human experience. Human experience with these natural toxicants has resulted in numerous
methods for addressing the risks presented by these substances (e.g. cooking, soaking or
avoidance). In general, ingestion of plant matter, including that containing natural toxicants, is
widely considered to have beneficial effects on human health in terms of general nutrition and
disease prevention.

Toxicants not known to adversely affect human health when consumed in high concentrations as
part of the normal diet , but with interesting properties or of general public interest (e.g. natural
remedies), are discussed below.

Capsaicin

General: Capsaicin is an alkaloid found in red peppers and in the spice cayenne, which is
derived from red peppers. This toxicant is the active ingredient which gives many foods their
hot taste.
12
Capsaicin works by decreasing the amount of substance P, a neurochemical found in synapses
responsible for transmitting pain signals. It is often used as a natural remedy for arthritis and
general muscle soreness (in creams of 0.025 or 0.075%). Capsaicin creams have also proven to
be effective in relieving secondary effects (e.g. itchiness and redness) associated with psoriasis52.
It is one of two toxicants (psoralens being the other class) which can penetrate the skin, although
this is greatly facilitated by the vectors in which the capsaicin is dispersed.
(Capsaicin is easily dispersed in fats and oils, which have good dermal penetration
characteristics.) Lethal toxic levels of purified crystalline capsaicin have been determined at 190
mg/kg for consumption and 521 mg/kg when applied topically (lethality is presumed to be due
to respiratory paralysis). It is estimated that a minimum oral lethal dose for an average person of
150 lbs (68 kg) would be 2.2 kg of capsaicin (dry pepper weight, by consumption)53.
Extrapolation from the mouse model suggests that a lethal dermal exposure for a 150 pound
individual would be 5.94 kilograms54.

Regulatory Status: Capsaicin is exempt from requirement for a tolerance for use on food crops
(40 CFR 180.1165). It has also been approved for use by the EPA for use as a biopesticide
repellent55.

Current Testing Methods: Capsaicinoids can currently be detected by reverse-phase HPLC56.

The method referenced here was mentioned by the American Society for Testing and Materials
(ASTM) in their publication; E-1083-88: Standard Test Method for Sensory Evaluation of Red
Pepper Heat57.

52
Ellis, C.N., et al. A Double-Blind Evaluation of Topical Capsaicin in Pruitic Psoriasis. J Amer Acad Derm
29:438-442. 1993.
53
Glinsukon, T., Stitmunnaithum, V., Toskulkao, C., Buranawuti, T., and V. Tangkrisanavinont. Acute Toxicity of
Capsaicin in Several Animal Species. Toxicon. 18(2):215-20. 1980.
54
Capsaicin Technical Fact Sheet. National Pesticide Information Center.
http://npic.orst.edu/factsheet/Capsaicintech.pdf.
55
US EPA Reregistration Eligibility Document. EPA-738-F-92-016. 1992.
56
Hoffman, P.G., Lego, M.C., and W.G. Galetto. Separation and Quantitation of Red Pepper Major Heat Principles
by Reverse-Phase High Pressure Liquid Chromatography. J. Agric. Food Chem. 31:1326-1330. 1983.
57
http://www.astm.org

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