What is annatto extract

No control of food intake was made after 6 h Levy et al. The average values and range of concentrations of bixin and norbixin in the plasma of the subjects are shown in Table 3. Table 3. Plasma concentrations of bixin and norbixin in volunteers given a single dose of commercial annatto food colour. Average Range Average Range 0 2. At the termination of the day studies of toxicity with annatto extracts B, E, and F conducted and reported later, liver samples were taken from 10 males and 10 females in the groups with the highest numbers of animals Bowman Research, b.

Microsomes prepared from these samples were frozen, transported, and analysed for the expression of CYP apoproteins using highly specialized Western blotting techniques by which levels of specific apoproteins can be assessed Boobis, In all cases, the data were consistent with those reported in previous publications. None of the extracts induced CYP2E1 expression. In comparison with the positive control pregnanolone 16 alpha- acarbonitrile, PCN , any PCN-type induction by the annatto extracts was extremely modest.

There was an appreciable difference between the sexes, females exhibiting a much weaker response, consistent with published data. Hepatic microsomal CYP4A was constitutively expressed in both male and female rats, levels being four to fivefold higher in males. There was no increase in the expression of CYP4A in the females receiving low doses of annatto F, but in the females receiving high doses, there was a five to six-fold increase in expression. Similarly, annatto B caused a dose-dependent increase in the expression of CYP4A in male rats, whilst any increase in females was very modest.

The effects of annatto E were less consistent, with the expression of CYP4A in males increasing by approximately three-fold, apparent only in the group receiving the higher dose. In females, the response was also not as consistent, but increased expression of CYP4A was observed in groups receiving both the low dose approximately three-fold and the high dose approximately two-fold.

The positive control, clofibric acid, produced the anticipated increase in the expression of CYP4A. The increases in expression of CYP4A in male rats after treatment with high doses of the annatto extracts, particularly annatto F, were similar to those observed after treatment with clofibric acid Boobis, Bixin has also been investigated for its effect on drug-metabolizing enzymes.

Control animals received diet only or the enzyme-inducing agent, 3-methylcholanthrene. CYP enzyme activity, glutathione-S-transferase and reduced glutathione status, and carotenoid uptake into the tissues were all assessed. The pharmacological activity of extracts of the root of Bixa orellana has been considered in previous evaluations by the Committee.

Since then, a few observations have been made concerning the effects of annatto preparations obtained from the seeds of Bixa orellana on the physiological and biochemical functioning of the body after administration to animals.

A suspension of the seeds of Bixa orellana in oil, known as "bush tea", is used in the West Indies as a folk remedy for the treatment of diabetes mellitus. Six experiments were performed on each animal, to determine the effect on a 3-h oral glucose tolerance test. Both types of extract oil or ethanol produced a mild suppression of the rate of increase of plasma glucose at the lower dose, and at the same time depressed plasma insulin concentrations, but not significantly.

In a subsequent study, the ethanol extract previously shown to cause hyperglycaemia was dried to produce a reddish-brown crystalline solid that was redissolved as a neutral aqueous solution. Oral glucose tolerance tests performed immediately and at weekly intervals, confirmed the hyperglycaemic action of this extract of annatto, which persisted for 4 weeks after treatment had ended. Concomitant administration of riboflavin 3 mg per day to some animals greatly reduced the hyperglycaemic effect.

Examination by electron microscope of the liver and pancreas of treated animals revealed disarrangement of internal mitochondrial structure, fusion of mitochondria and formation of a large number of residual bodies.

The apparent stacking of the hepatic mitochondrial cristae was also a striking feature. Dogs treated additionally with riboflavin exhibited only minimal mitochondrial changes Morrison et al. This revealed that the active constituent responsible for the hyperglycaemic action was the methyl ester trans- bixin relative molecular mass, As little as 0. The purified material also caused mitochondrial damage in the liver and pancreas but, as with the crude extract, pre-treatment of the dogs with riboflavin a few days before the trans- bixin was administered resulted in little or no damage Morrison et al.

This hyperglycaemic activity has been confirmed in rats, but an hypoglycaemic effect has been reported in a study in mice. The authors showed that when rats were given norbixin in their drinking-water for 21 days, hyperglycaemia ranging from On the other hand, in mice given norbixin, hypoglycaemia ranging from The pancreatic beta- cells were functional, as indicated by the presence of hyperinsulinaemia in rats and hypoinsulinaemia in mice treated in this way Fernandes et al.

Together with 16 other Guatemalan medicinal plants, Bixa orellana was investigated for ability to prevent platelet aggregation. Bixa orellana , and a number of traditional remedies, inhibited the thrombin-induced aggregation of washed human platelets Villar et al.

The authors concluded that these food colours might regulate the production of immunoglobulins Kuramoto et al. Bixin, together with other carotenoids such as beta- carotene, lutein and canthaxanthin, has been shown to suppress the respiratory burst induced by paramethoxyamphetamine PMA in rat peritoneal macrophages Zhao et al. The action appears to be associated with the ability of carotenoids to scavenge superoxide, and the authors suggested a protective role for carotenoids in vivo to protect host cells from the harmful effects of oxygen metabolites see section 2.

Earlier work showed that bixin binds to the non-polar regions of mitochondria thought to be associated with high-energy states Inada et al.

Furthermore, bixin acts as an inhibitor of the ATP-forming process state 3 associated with mitochondrial respiration Hirose et al. The effects of annatto on cellular enzymes and metabolites in the rat have also been investigated as part of a wider investigation of synthetic and natural food colours, in an attempt to provide a simple diagnostic test for toxicological damage.

Four groups of eight male albino rats were used for the part of the study concerning annatto. Animals in groups 2 and 3 were killed by decapitation 24 h after the last dose, while the animals in group 4 were killed 1 week after the cessation of treatment.

Brain, liver and kidneys were removed from all animals, and cytoplasm and mitochondria were prepared for measurement of the activities of glucosephosphate dehydrogenase GPD and 6-phosphogluconate dehydrogenase 6-PGD.

Annatto produced a minimal rise in the activity of these enzymes, mainly in the liver, and consistently less than that produced by synthetic food colours such as tartrazine, carmoisine and sunset yellow. In group 4 , in which treatment had been stopped for a week before killing, the enzyme activities had returned to normal. It was noted that these effects could be a result of metabolism of the colour within the body Hamama et al. As previously, these effects were much smaller than those seen with synthetic food colours, and normal values rapidly returned after treatment was stopped Hamama et al.

Studies of acute toxicity in rats and mice were reported previously Annex 1, references 35 , More recently, Germano et al. The acute toxicity of annatto when administered orally is low. Studies of toxicity in mice, rats, dogs and pigs have been previously reported; the results for all four species were discussed in the previous evaluations by the Committee. Relevant results are summarized below, together with the results of new studies.

No signs of toxicity were detected by histopathological analysis of the liver, and no enhancement in DNA strand breakage was detected in liver or kidney from mice treated with annatto pigments, as evaluated by the comet assay.

Hypoglycaemia was induced in the mice as discussed in section 2. Five groups of six male and six female Wistar rats received diets containing three annatto preparations, either singly or in combination, as follows:.

Liver, kidneys and abdominal adipose tissue were removed at postmortem, weighed, examined gross appearance only and subjected to an extraction procedure to identify the presence of annatto pigments. Body weights and food intakes were similar in all treatment groups and not significantly different from those of the control group.

The appearances and weights of organs and tissues showed that the animals were in good health, with no differences between the groups. Analysis of the extracts of liver, kidney, adipose tissue and carcass failed to reveal the presence of annatto pigments Philp, No toxic effects on plasma clinical chemistry parameters were detected.

A significant hyperglycaemic effect was reported in contrast to the hypoglycaemia reported in mice ; this is discussed in section 2. In a day study of palatability, groups of five males and five females were fed diets containing varying amounts of annatto extract.

Body weights were recorded twice weekly and food intakes were measured weekly. After 28 days of treatment, the animals were killed and subjected to full postmortem examination, major organs were weighed and tissues were fixed for histological examination. It was concluded that annatto B was well tolerated in the diet at concentrations of up to 50 ppm; no dose-related absolute or relative organ weight changes were observed, but macroscopic examination revealed orange coloration of the gastrointestinal tract.

At the highest dose, orange coloration of the mesentery was observed in a few animals. Annatto D was well tolerated in the diet at concentrations of up to 11 ppm. Increases in concentration up to 22 ppm were less well accepted and there was a suggestion that gradual increase in dosage was better tolerated than abrupt introduction of a high dose. An increase in relative liver weights was recorded for all treated males compared with their respective controls, but only in females treated with up to 22 ppm.

Macroscopic examination again revealed orange coloration of the tongue and gastrointestinal tract in the majority of the treated animals. In addition, the mesentery tissue was stained orange in several of the treated animals. Annatto E at concentrations of up to and including 40 ppm was found to be acceptable to the animals. However, increased relative liver weights were seen in all treated animals. Microscopic examination of the livers revealed diffuse and periacinar hepatocyte hypertrophy in rats treated with 20 —30 ppm or 20 —40 ppm over the period of study.

Again, orange staining of the tongue and gastrointestinal tract was reported for all animals receiving annatto E. The majority of the treated animals had orange staining of the mesentery. Treatment with annatto F resulted in orange coloration of the tongue and gastrointestinal tract in the majority of the animals and of the mesentery in a few animals. The most significant finding in this study was the increased relative liver weight seen in all groups treated with annatto F at concentrations ranging from to 20 ppm.

Animals treated with annatto G showed similar orange coloration of the tongue and gastrointestinal tract to that in animals treated with annatto F. The four materials tested comprised two preparations of bixin annatto B and annatto E and of norbixin annatto C and annatto F , one with an high pigment content and one with a lower pigment content.

In addition, a functional observational battery of tests was performed on animals fed with annatto B and F. As the sample of annatto B used in the day study contained high levels of residual solvent, a new sample of annatto B was obtained from the manufacturers. The samples of annatto E and annatto F used in the day studies were from the same stock as that used in the day studies. There was no overall significant effect on body-weight gain in either the males or females.

The functional observational battery tests revealed no treatment-related effects on behavioural, motor or neurological activity. Ophthalmoscopic examinations revealed no abnormalities.

Haematological investigations revealed no findings that were clearly related to treatment. Raised lymphocyte counts 9. When compared with the controls, statistically but not biologically significant elevations were seen in plasma concentrations of phosphorus 2. This effect was not seen in males. These findings were not accompanied by histopathological evidence of liver damage and were considered by the author to be of only minor importance in the evaluation of the toxicity of annatto B. Minor statistically significant variations in albumin:globulin ratios were observed, but these were not dose-related, and were not considered to be toxicologically significant.

Significantly raised concentrations of protein were recorded in urine samples obtained from males receiving annatto B at a concentration of 50 ppm. Microscopic examination of the urine sediment revealed the presence of "crystals with a red sediment on top" for one female at ppm and two males at 16 ppm.

In another female at ppm and in three males and four females at 50 ppm, patches of red staining were seen on the microscope slides. These presumably reflect the dark colour of the annatto extract. These increases were not associated with any histopathological findings. No other effects on absolute or relative organ weights were observed.

Four groups of the same strain of rats received diets containing annatto E at a concentration of 0, , 10 or 30 ppm for 90 days. The administration of annatto E produced a number of treatment-related changes, particularly in the groups receiving the highest dose. At the intermediate dose, a reduced body-weight gain in the absence of any effect on food conversion efficiency in the females was most likely to have been caused by an effect of the palatability of the diet on food intake.

Increased liver weights seen in many of the treated animals, with the exception of females at ppm, were associated with slight adaptive centrilobular hepatocyte hypertrophy; no clear signs of liver pathology were noticed. Thyroid follicular cell hypertrophy was present in some animals receiving annatto E at a concentration of 30 ppm. Raised plasma concentrations of creatinine were seen in all females treated with annatto E and in males at 30 ppm.

A slight increase in the weight of the kidney relative to body weight was seen in the females at the highest concentration. Raised plasma concentrations of phosphorus seen in males and females at the highest concentration of annatto E may indicate a reduction in the rate of glomerular filtration. The no-observed-adverse-effect level NOAEL for both sexes can be considered to be the intermediate concentration of 10 ppm in the diet. Four groups of the same strain of rats received diets containing annatto F at a concentration of 0, , or ppm for 90 days.

Since annatto F contains The administration of annatto F did not result in any treatment-related deaths. Transitory reductions in body weight and food consumption were noticed in males receiving the highest dose of annatto F in the first weeks of the experiment.

The functional observational battery tests revealed no evidence of behavioural, motoror neurotoxicity, and ophthalmoscopic examination did not indicate the presence of any treatment-related lesions. Slightly but statistically significant low erythrocyte volume fractions, concentrations of haemoglobin, red blood cell counts and mean cell volumes were apparent for females at or ppm, suggesting slight anaemia caused by annatto F.

In the animals receiving annatto F at a concentration of ppm, significantly increased activity of alkaline phosphatase was reported in males, with activity of alanine amino-transferase also being increased in two of these males.

Concentrations of urea, creatinine, glucose, triglyceride and albumin were raised in males and females at ppm. Concentrations of total protein were raised in females. Reduced concentrations of alpha- 1-globulin and beta- globulin were noticed in males, but concentrations of alpha- 1-globulin were raised in females. The albumin:globulin ratio in males was increased.

At ppm, total protein and concentrations of albumin were significantly raised in females, whereas the concentration of beta- globulin and the albumin:globulin ratio were increased in males. The plasma obtained from all treated animals had a definite yellow coloration, which was presumably norbixin.

A very marked increase in absolute and relative liver weights were seen in males treated with annatto F at a concentration of ppm and in females at and ppm. In females at ppm, only slight, although statistically significant, increases were noted. No changes were found microscopically, except in animals receiving a concentration of ppm, in which these changes were associated with centrilobular hepatocyte hypertrophy.

The hypertrophy observed after 90 days of treatment was no greater than that observed after 28 days of treatment in the study on annatto F reported above. The weight of the kidney was slightly increased in males and females at ppm and in males at ppm.

Plasma concentrations of creatinine were raised in animals receiving annatto F at a concentration of or ppm, and concentrations of urea were raised in males and females at ppm. I Inc. Absolute and relative liver weights were increased and mild hypertrophy and mild necrosis of hepatocytes were noticed on histopathological examination of rats treated with annatto C.

Electron microscopic examination revealed an abundance of mitochrondia in the cytoplasm of hepatocytes of the treated animals. It was concluded that the hypertrophy of hepatocytes was linked to the increase of cytoplasmic mitochondria SanEi F. There was no effect on body weight or food intake, and no changes in haematology parameters.

Changes in clinical chemistry parameters were observed in animals at the highest dose and, to a minor degree, at the intermediate dose. Increased absolute and relative liver weights were observed in animals of each sex in the groups receiving the two higher doses, and this was accompanied by hepatocyte hypertrophy in animals in these groups and focal necrosis in one male and one female in the groups receiving the highest dose.

A small increase in kidney weight in each sex at the highest dose was not accompanied by any histopathological changes and may reflect a physiological response to the extra metabolic load imposed by the annatto extract. Three studies in mice and three studies in rats were previously submitted for evaluation. No new studies have become available. No separate studies of carcinogenicity, other than those submitted for previous evaluations, have been conducted.

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What it is Benefits Uses Safety Bottom line Annatto is a type of food coloring made from the seeds of the achiote tree Bixa orellana. This article reviews the uses, benefits, and side effects of annatto. Share on Pinterest. What is annatto? Potential health benefits of annatto. Annatto uses.

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Top 10 Healthy Cuisines from Around the World. Medically reviewed by Amy Richter, RD. Annatto and its extracts are now widely used in an artisanal or industrial scale as a coloring agent in many processed food products, such as cheeses, dairy spreads, butter and margarine, custards, cakes and other baked goods, potatoes, snack foods, breakfast cereals, smoked fish, sausages, and more.

In these uses, annatto is a natural alternative to synthetic food coloring compounds, but it has been linked to rare cases of food-related allergies. Annatto is of particular commercial value in the United States because the Food and Drug Administration considers colorants derived from it to be "exempt of certification". Food R G B is Annatto food color manufacturer - natural yellow food color manufacturer.

We carefully choose the best seeds of the Bixa orellana, then extract natural yellow color. Our special manufacturing process removes odor and flavor from the annatto.