Beetroot red (E 162) is a natural colour containing a number of pigments, all belonging to the class known as betalains. The main colouring principle consists of a number of betacyanins. The EFSA was asked for a re-evaluation.
The Panel was not provided with a newly submitted dossier and based its evaluation on previous evaluations, additional literature that has become available since then, and data available following a European Food Safety Authority (EFSA) public call for data. The Panel noted that not all of the original studies on which previous evaluations were based were available for this re-evaluation.
To assist in identifying any emerging issue or any information relevant for the risk assessment, EFSA outsourced a contract to deliver an updated literature review on toxicological endpoints, dietary exposure and occurrence levels of beetroot red (E 162), which covered the period up to the end of 2014. A further update has been performed by the Panel.
Beetroot red (E 162) is a natural colour authorised as a food additive in the European Union (EU) in accordance with Annex II to Regulation (EC) No 1333/2008. It has been previously evaluated by the Scientific Committee for Food (SCF) in 1975 and in relation to special medical purposes for young children in 1996 (SCF, 1975, 1997a). The Joint FAO/WHO Expert Committee on Food Additives (JECFA) evaluated beetroot red in 1974, 1978, 1982 and 1987 (JECFA 1975, 1978, 1982, 1987, 1988). A previous JECFA temporary Acceptable Daily Intake (ADI) ‘not specified’ was withdrawn in 1982 due to the fact that information on metabolism and long-term toxicity that had been requested by JECFA was still not available. Neither the SCF nor JECFA have currently specified an ADI.
Beetroot red is a natural colour obtained from the roots of natural strains of red beets (Beta vulgaris L. var. ruba). The most commonly used synonyms for beetroot red in the published literature are Beet Red, Betanin, INS No. 162.
Beetroot red (E 162) contains a number of different pigments, all belonging to the general class known as betalains. The main colouring principle consists of a number of betacyanins (red), of which betanin accounts for 75–95% and isobetanin (epimer of betanin) 15–45%; a range of other betacyanins can also be detected, amounting up to 20% of the total content of betacyanins. Vulgaxanthin I (25–70%) and vulgaxanthin II (5–15%) are present in the betaxanthin (yellow) as well as several degradation products of betalains (light brown). Besides, the colour pigments beetroot red contains sugars, salts and/or proteins naturally occurring in red beets.
A number of different beetroot red products may be marketed, ranging from press juices or aqueous extracts of shredded roots to more concentrated or refined forms including pastes, powders and other solid forms. The Panel noted that the current re-evaluation does only refer to beetroot red prepared by pressing crushed beet as pressed juice or by aqueous extraction, in accordance with the definition of the Commission Regulation (EU) No 231/2012 and not to preparations manufactured by solvent extraction with methanol or ethanol.
Specifications for beetroot red have been defined in Commission Regulation (EU) No 231/2012 and by JECFA (2006). EC specifications including purity criteria for beetroot red define not less than 0.4% of the commercial material must be betanin pigment. The remaining 99.6% is accounted for by sugars, salts and proteins naturally occurring in red beets, and a small amount of other pigments belonging to the class of betalains and but this is not further specified. According to industry (NATCOL, 2012), non-colouring substances present in beetroot red are sugars, proteins, minerals, organic acids, vitamins, sterols, purines and phenolic compounds. The Panel noted that the reported content of betanin in beetroot to be around 0.4%. The Panel further noted that the specification for the content of red colour (expressed as betanin) in beetroot red, as not less than 0.4%, may give rise to some confusion, given the number of different forms of beetroot red that may be on the market, including simple extracts, refined extracts and spray–dried powders. The Panel also noted that some forms of beetroot red designated as colouring foodstuffs rather than as food colours may contain more than 0.4% colouring matter. The Panel considered that revision of the current specification to reflect betanin content on a dried solids basis could be appropriate.
The specifications for beetroot red include a maximum level for nitrate of < 2 g nitrate anion/g of red colour, given the relatively high content of nitrate in beetroot. The Panel considered a nitrate limit important in minimising the contribution to nitrate intake from this source as a food additive. This should be evaluated together with dietary exposure and use as a colouring food when evaluating the total intake of nitrate by the population.
There are no validated methods for the analysis of beetroot red (or betanin) in foods that may be used for official purposes.
The toxicological studies are described variously as using (1) red beet juice, (2) beetroot red, (3) betalains, (4) betanin, (5) beetroot red extracts, (6) freeze-dried beetroot, (7) beet red powder. All of these products inevitably contained betanin pigment. However, the Panel considered that only those (relatively few) studies carried out with a test material containing not less than 0.4% betanin are relevant for the assessment of the safety of the food additive beetroot red (E 162). It is unclear which of the above substances most closely resembles the commercial food colour and whether testing of purified betanin provides sufficient insight into the biological and toxicological behaviour of the commercial product.
The betacyanin pigments from beetroot red are metabolised or degraded in vitro by chopped tissue preparations from the stomach, small and large intestine, indicating that some breakdown of pigments is likely to occur in the gastrointestinal tract following oral administration of beetroot red. Studies in humans, supported by animal studies, have however shown that the betalain pigments present in beetroot are absorbed in an intact form to a limited extent (approximately 3% of the dose in rats and less than 1% of the dose in humans) after oral administration and are not metabolised further in the body, as demonstrated by the excretion of betanin, isobetanin and other betacyanin pigments at low levels in urine. Information from intravenous and intraperitoneal administration of beetroot extracts in rats demonstrated that intact pigments were extensively excreted unchanged in the urine. In humans, ingestion of beetroot can produce red urine (‘beeturia’) in some individuals. It has been suggested that beeturia is more a function of an individual’s physiological constitution than a phenomenon under direct polymorphic genetic control.
The Panel noted that toxicological studies carried out on material conforming to the specifications for beetroot red are limited in number. However, there was no evidence of adverse effects in a range of studies conducted with poorly defined material and/or judged to be of limited relevance for the assessment of beetroot red (E 162).
The genotoxic potential of beetroot red (E 162) cannot be evaluated based on the available data.
There are only limited or inadequate studies available on the carcinogenicity of beetroot red and therefore the Panel could not conclude on the chronic toxicity and carcinogenicity of beetroot red.
No adequate studies on reproduction and developmental toxicity were available.
There is no indication of intolerance or allergenicity of beetroot red in the available literature.
Exposure assessments of food additives under re-evaluation are carried out by the ANS Panel based on (1) maximum permitted levels (MPLs) set down in the EU legislation (defined as the regulatory maximum level exposure assessment scenario) and (2) usage or analytical data (defined as therefined exposure assessment scenario). It was not possible to carry out a scenario based on the MPLs set out in EU legislation, as, for all food categories, beetroot red (E 162) is authorised according to quantum satis (QS). However, maximum levels of the available data were used to provide a conservative estimate scenario (noted as the maximum level exposure assessment scenario). With regard to the refined exposure assessment scenario, only reported use levels were made available by industry. The Panel considers that the refined exposure assessment approach results in more realistic long-term exposure estimates because of the underlying assumptions and the concentration data used.
Reported use levels were all provided in mg betanin/kg food. Usages notes were also added by NATCOL (NATCOL, 2015) which could mention the percentages of food items per food category in which the food additive (E 162) or the colouring food (CFS) is used. These percentages were not taken into account as they should have been used to reduce the number of time the additive E 162 is used. This is not possible in the current modelling. Meanwhile, the reported use levels provided by NATCOL are the correct ones when the food additive E 162 is used. Therefore, the scenarios presented in the current opinion assume that irrespective of whether the food additive or colouring food is used, all the betanin is coming from the food additive E 162.
The Panel noted that the refined exposure estimates will not cover future changes in the level of use of E 162.
Using the maximum level exposure assessment scenario, mean exposure to beetroot red (E 162) from its use as a food additive ranged from 0.1 mg/kg bw/day for the elderly to 2.1 mg/kg bw/day in toddlers, whereas the high exposure using this scenario ranged from 0.3 mg/kg bw/day for the elderly to 3.6 mg/kg bw/day in children. Using the refined brand-loyal assessment exposure scenario, mean exposure to beetroot red (E 162) from its use as a food additive ranged from 0.1 mg/kg bw/day in adults and the elderly to 1.6 mg/kg bw/day in toddlers. The high exposure to beetroot red (E 162) using this scenario ranged from 0.2 mg/kg bw/day in the elderly to 2.8 mg/kg bw/day in toddlers. Using the refined non-brand-loyal assessment exposure scenario, mean exposure to beetroot red (E 162) from its use as food additive ranged from 0.05 mg/kg bw/day for the elderly to 1.0 mg/kg bw/day in toddlers. The high exposure to beetroot red (E 162) from its use as food additive using this scenario ranged from 0.1 mg/kg bw/day for the elderly to 1.8 mg/kg bw/day in infants. Overall, the lowest exposure to beetroot red (E 162) was estimated for the elderly, whereas the highest exposure to beetroot red (E 162) was calculated for toddlers in all scenarios. The food categories that, at the individual level, had the highest contribution to the total individual exposure to beetroot red (E 162) were breakfast cereals, fine bakery wares, soups and broths, and flavoured drinks.
Mean intakes of betanin from the regular diet for consumers only are in the range of the mean estimated exposure from the use of the food additive itself (Table 4, non-brand loyal consumer scenario).
The Panel concluded that the currently available toxicological database was inadequate to establish an ADI for beetroot red as defined by the specifications set for the food additive E 162. However, the Panel concurred with SCF opinion that ‘for colours for which an ADI cannot be established… exceptions might be made in the case of compounds which are in fact constituents of food and derived from coloured natural foods by purely physical process’ (SCF, 1975).
The colouring principles in E 162 are natural dietary constituents having a long history of food consumption. In addition, the betanin exposure resulting from the use of beetroot red (E 162) as food additives is in the same range as the exposure to the betanin from the regular diet. Therefore, the Panel concluded that, at the reported use levels, beetroot red (E 162) is not of safety concern as regards its current use as a food additive.
The Panel recommended that:
- revision of the current specification to reflect betanin content on a dried solids basis could be appropriate;
- limits for mycotoxin contamination may be relevant for the specifications of beetroot red (E 162);
- maximum limits for the toxic elements (arsenic, lead, mercury and cadmium) present as impurities and nitrates in the EC specification for beetroot red (E 162) should be revised in order to ensure that beetroot red (E 162) as a food additive will not be a significant source of exposure;
- EU Regulation should include the specification for solubility as given in the JECFA specification.