norbixin found that doses of 0.8, 7.5, and
68 mg/kg produced hyperinsulinemia in rats
and that doses of 56 and 351 mg/kg produced
hypoinsulinemia in mice. In the rats, the
extract also caused a significant decrease in
globulins and plasma total protein levels
without evidence of any adverse effects on
the liver or plasma chemistry.^14 Adminis-
tered to pregnant rats, annatto food color
containing 28% bixin caused no adverse
effects to the mothers and none to the fe-
tuses. A developmental and maternal No-
Observed-Adverse-Effect-Level (NOEL) for
the extract was proposed as 500 mg/kg p.o.
per day or greater and for bixin, at least
140 mg/kg p.o. per day.^15
At high oral doses in rats, norbixin (8.5
and 74 mg/kg) produced hyperglycemia.^14
However, in a subchronic (2-week) toxicity
test in rats of either sex, the NOEL of
norbixin was 0.1% of the diet (76 and
69 mg/kg p.o. for males and females, respec-
tively). At 0.3% and 0.9% of the diet, nor-
bixin produced a pronounced elevation in
liver weights, hepatocyte hypertrophy, and
in blood work, increases in albumin/globulin,
albumin, alkaline phosphatase, total protein,
and phospholipid ratios.^16 In the micronucle-
us test in male mice, annatto food colorant at
dietary levels of 1330, 5330, or 10,670 ppm
for 7 days failed to produce any mutagenic
effects. However, the highest concentration
increased the mutagenic effect of a mutagen
(cyclophosphamide).^17 Similar results were
found fromin vitrotests in which low con-
centrations of norbixin (10–50mM) protected
DNA against oxidative damage, while at
higher concentrations it enhanced oxidative
damage to DNA.^18
Inductionof drug-metabolizingenzymesof
the liver (CYP2B and CYP1A) was found
from oral dosing of female rats with an extract
of the seeds containing 28% bixin; 95% pure
bixin showed only weak inducing activity.^19
Human allergic reactions to annatto food
coloring include IgE-mediated anaphylaxis in
a man who consumed a breakfast cereal con-
taining the dye. Other clinical reports suggest
the possibility that angioedema and urticaria
occur as allergic reactions to the dye in some
individuals.^20USESThe current major commercial uses of annatto
colors are in foods, especially dairy;^21 also
used in drugs as color coatings for granules,
pills, tablets, and herbal preparations, as well
as in cosmetics (e.g., lipsticks) and hair dyes.
Aqueous alkaline extracts are extensively
used in coloring cheeses (especially cheddar)
and to a lesser extent in ice creams and other
dairy products. Oil-soluble extracts are used in
oily food products such as salad oils, popcorn
oil, butter, margarine, and sausage casings.
Major food categories in which annatto color
is used include alcoholic and nonalcoholic
beverages, frozen dairy desserts, baked goods,
meat and meat products, condiments and
relishes, fats and oils, snack foods, and grav-
ies, with highest average maximum use level
reported for the extract in baked goods (ca.
0.24%).Traditional Medicine. The seeds have been
used in treating tumors of the oral cavity, as a
purgative, antipruritic,^8 and in the treatment
of venereal diseases. The pulp that surrounds
the seeds is topically applied as a mosquito
repellant.^22 Preparations of the leaves are
traditionally used in treating gonorrhea,^23
nausea, as a gargle for sore throat, and for
oral hygiene.^8COMMERCIAL PREPARATIONSCrude, oil- and water-soluble extracts and
spray-dried powders.
Also available for domestic use as ground
or whole seeds in supermarkets or ethnic
stores in metropolitan areas in the United
States.Regulatory Status. Approved for use as a
colorant in foods (§73.30), drugs
(§73.1030), and cosmetics (§73.2030).Annatto 41