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[1] One of the most effective argument to play down the danger of cancer is to classify the presence of a contaminant as not relevant because of the no-effect-level.
According to that theory the reduction of the amount of contaminants leads to a point where a carcinogenic activity only develops after 100 to 150 years,that is after dying from other causes.

Unfortunately cancer comes up sooner, for the time being 25% of all human beings die of cancer.
More as 50% of all cases are caused by ecological factors.
The theory of no-effect-level can only be applied on persons who wish not to reproduce himself.

The genes who suffered mutations because of ecological factors should not be passed on to following generations in order not to pass on the genetic predisposition to cancer.
Carcinogenic activities are often analysed considering an isolated cause or a single agent.

Today there are lots of chemicals with carcinogenic activity boosting the effects in a synergistic way. Examples of such synergistic activities are:

Nitrosamines with PCB, benzpyrene BHT and traces of mercury.
The international institute of cancer in Lyon, France has proved that damages on the structure of chromosomes caused by toxic substances are passed on from generation to generation.

The institute gave small dose of nitrosamines to pregnant mice. Typical tumours were not only found on the mother but although on following generations who had no contact with nitrosamines.
According to professor Schmähl, researcher on cancer in Heidelberg,Germany, there is no dose, even being very small, that is free of danger to cause cancer because of the effect of synergistic addition of the effects.
The fact that a mouse can live with a small dose of carcinogen agents does not prove anything.

The mouse does not smoke, it does not breathe sulphur dioxide, it does not take medicine, it does not eat ham, smoked salmon or hamburgers. Therefore we have to observe very critically all additives and all ingredients of our food. [2]

Additives in the European Food Law and the Codex Alimentarius

The European food law is more stringent as the Codex Alimentarius. If a food or any agricultural product meets European rules it is also fit for Codex. The Codex used the principles of the EU food law. To instance, the E-Numbers of additives from the European numbering system are being used by the Codex deleting the E in front of the number.

Additives in brief

The Regulation of food additives in Europe changed. 1333/2008 will be in force. However, the lists of additives of older directives (94/35/EC, 94/36/EC and 95/2/EC) remain valid because new lists are not ready yet.

Additives in the European Union

The article of the European Commission on Food Additives and flavourings gives a summery of European food law.

Regulation 1333/208 on food additives

The Regulation brings together in a single legislative act all types of food additives including colours and sweeteners.
Food enzymes are covered by Regulation (EC) No 1332/2008.

List of additives

Whilst these lists cited as "Annexes" in 133/2008 are being drawn up, the Annexes of Directives 94/35/EC, 94/36/EC and 95/2/EC remain in force.

Codex Food additives

The "Codex General Standard for Food Additives" (GSFA, Codex STAN 192-1995) sets forth the conditions under which permitted food additives may be used in all foods, whether or not they have previously been standardized by Codex.

Contact between the European Commission and the Codex Alimentarius

The Codex Alimentarius Commission adopts standards, codes of practices and other related texts that are prepared by specialised Codex Committees and Ad hoc Task Forces. The 27 Member States of the European Union are all members of the Codex Alimentarius Commission. In 2003, the European Community (now the EU) also became a full member of the Codex Alimentarius Commission and shares the competence with its Member States on the basis of the level of harmonisation of the relevant legislation. Since the entry into force of the Treaty of Lisbon on 1 December 2009 the European Union replaced the European Community. The EU and its Member States elaborate EU position papers on issues discussed in the Codex Alimentarius Commission, the various Codex Committees and Task Forces.

European regulation

All directives are available at:
Year and Number of the Directive Old directives cite only the two last digits of the year. New directives give four digits such as: 91/414 = year 1991 Nr.414 and 2009/128 = year 2009 Nr.128
To search by one word Just type a word, such as "pesticides"

Codex Alimentarius

The Codex Alimentarius Commission was created in 1963 by FAO and WHO to develop food standards, guidelines and related texts such as codes of practice under the Joint FAO/WHO Food Standards Programme. The main purposes of this Programme are protecting health of the consumers and ensuring fair trade practices in the food trade, and promoting coordination of all food standards work undertaken by international governmental and non-governmental organizations. [3]
- Current Standards: Here all Standards are available.
- Search system: Very efficient search engine of Codex
- Codex database for limits of pesticides in foods
- Codex database for veterinary drugs residues in foods
- Codex Food additives

Intensive farming in the European Union

Intensive farming bears the risk of overuse of antibiotics according the National Consumer Council March 11 1998 in London.
The council blames in the words of his Director Ruth Evans the European Unions common agricultural policy encouraging intensive farming.

Ruth Evans says:" The Common Agricultural policy is not just wasteful and costly to consumers,it also encourages farming practices which raise serious public health concerns.
Overintensive farming methods led to the BSE/CJD crisis.So long as we reward high output rather than high quality of food further risks are likely"
The Council enumerates in his report the following areas of concern:

1- Antibiotic

It is being used to treat , prevent and reduce disease and as a growth promoter in animals. The residues can be toxic and cause hypersensitivity to antibiotics in some humans.

Uncontrolled use of antibiotics increase the resistance of certain bacteria to these therapeutics. Salmonella and Escherichia coli are developing the capacity to resist the medicine which treats diseases on humans and on animals.

2- Genetic modification

The introduction of selected properties of an organism into another bears unknown risks which cannot be estimated by risk analysis.

3- Hormones

Hormones are used to promote animal growth.
This may lead to the development of sexual characteristics and certain cancers.
In spite of being forbidden in the European Community many hormones are illegal sold and used.


Nitrates are used to enrich the soil and promote growth to vegetable crops.
High use of nitrates are linked to the cyanosis called "blue baby Syndrome" and to stomach cancer.
The maximum nitrate levels in vegetables and water which have been established are sometimes exceeded.


Pesticides are used to protect crops and increase productivity.Exposure to certain pesticides may weaken the immune system and reduce resistance to cancer.

It has been noted that some pesticides are deposited in higher rates in the interior of fruits and vegetables as in the skin. Peeling and washing has therefore no effect in reducing the content of pesticides in food. According to Mrs. Evans the EU should promote the reduction of the use of antibiotics,pesticides and nitrates and strengthen the regulatory framework of the industry, which has proved to be inadequate.

In 1988 began in United Kingdom a monitoring of pesticide residues in food.
Since then there were no detectable levels of residues found in around 70% of examined samples.Only 1% exceeded the maximum residue levels.
Efforts however should be undertaken to achieve further reduction , because there is no-effect-level.

Red meat consumption and risk of cancer

The government of United Kingdom has made a statement that people who eat average 90 grams of cooked red meat per day, or around 8 to 10 portions a week need no reduction of red meat consumption.This includes beef, lamb and pork.
A lower consumption of red meat would probably reduce the risk of colon rectal cancer.
However eating to little meat could cause iron deficiency.
In order to correct wrong behaviour related to food it is recommended to:

Nitrates and nitrites are less detrimental to health as supposed

Gastrointestinal cancer and methemoglobinemia risk in infants may be increased by nitrates and nitrites in food. Hord, Tang an Bryan 2009, however, claim that the risks and health benefits associated with consumption of dietary nitrate and nitrite from fruits and leafy green vegetables should be re-evaluated. The authors stress that nitrates and nitrites from fruits and vegetables have a protective effect against coronary heart disease and ischemic stroke risk.

80 per cent of dietary nitrates are derived from vegetable consumption. The Dietary Approaches to Stop Hypertension (DASH) studies, using diets low in fat dairy products and 8-10 servings, lowered blood pressure to an extent similar to that achieved with medication. This was attributed to the high calcium, potassium and low sodium concentrations.

Hord and colleagues, however, say that the nitrate contained in some fruit and vegetables may be associated with the blood pressure reduction. Mammalian enzymes were found to have nitrate reductase activity as well bacterial nitrate reductases in mouth and gastrointestinal tract reduce nitrate to nitrite. The authors found a variability on nitrate concentration in vegetables, fruit, juice and fresh and processed meats, varying between 174 to 1222 mg in patterns of the DASH diets.

Risks associated with excessive nitrate and nitrite consumption

Hord and colleagues say that infants under six months exposed to excess nitrite may suffer from methhemoglobinemia and that infants fed home-prepared food are probably my be harmed by high nitrates in foods. Cured and processed meats with added nitrates have also a detrimental effect on health, but there are little evidences of cancer risk associated with cured meat.

The authors stress that any health risk from nitrates in fruits and vegetables are outweighed by their benefits, such as reduction of blood pressure. They call to reconsider the acceptable daily intake (ADI) levels for nitrites and nitrates and suggest to consider nitrates as nutrients.

EU food additives regulations

The regulation 1331/2008 [6] lays down a common procedure for the assessment and authorisation of food additives, food enzymes, food flavourings. It is therefore important for the procedure of approval of food additives, but not for the food industry and the consumer interested on positive lists of additives. These are given in 1332/2008 [7], 1333/2008 [8] and 1334/2008 [9] and the annexes Directives 94/35/EC [10], 94/36/EC [11]and 95/2/EC [12]. Feed additives 429/ 2008 should also be considered by animal breeders.

Transitional provisions for additives

In accordance with article 30 of Regulation (EC) No 1333/2008, additives that are permitted in food under Directives 94/35/EC, 94/36/EC and 95/2/EC and their conditions of use will be entered the Community list of food additives in Annex II to the regulation. To that end the compliance with their general and specific conditions of use must first be reviewed. This review should be completed by January 2011.

The use of food additives already permitted in Directives 94/35/EC, 94/36/EC and 95/2/EC will continue to be permitted until the review has been finalised and the additives have been transferred in the Community list of food additives in the Annex II of the new regulation. In the mean time, when necessary, until the establishment of the Community list of food additives, the annexes to the current directives can be amended based on a comitology procedure.

European food colours legislation

All colours which are allowed to be added to foods are included in positive lists. Substances therein have been tested for their safety and meet specific purity criteria.

The Colour Directive 94/36/EC gives the actual permitted food colours. The safety of food colours and other food additives is evaluated by the Scientific Committee on Food (SCF), an advisory expert committee of the European Commission, located in Brussels.

The EU Commission Directive 95/45/EC of 26 July 1995 lays down specific purity criteria concerning colours for use in foodstuffs with amendment Directives 1999/75/EC, 2001/50/EC and 2004/47/EC.

The European Commission regulations are binding to all member countries of the EU, and have to be implemented into their national laws. Non-EU member states food additives are regulated by their national authorities.

World Health Organisation

The WHO and the FAO members of the UN have an advisory committee, the Joint WHO/FAO Expert Committee on Food Additives (JECFA), The European SCF and JEFF A's toxicological evaluation, an ADI (Acceptable Daily Intake) value, expressed in mg/kg body weight per day, is allocated to the additive, or its use is not recommended.

The E numbers for colour additives range from E100 (curcumin) to E180 (lithorubine BK). Since a pigment derived from extraction may be differently specified as the same pigment derived from chemical synthesis, it may be characterised by a differentiated E number: beta- carotene extracts e.g. are listed under E160ai (mixed carotenes), and synthetic beta-carotene under E 160aii (beta-carotene).

Food additive colours are defined in this regulation as: [11]
"Substances which add or restore colour in a food, and include natural sources which are normally not consumed as a foodstuff as such and not normally used as a characteristic ingredient in food. Thereby, the Colour Directive excludes colouring foodstuffs and food ingredients, which may be used in the preparation of a final food, from the food additive regulation, such as tomato juice (lycopene) or red beet juice (betanine) which is added to some foods, colouring them.

According to the Colour Directive, the legal situation of such pigments changes, when they are selectively extracted -relative to the nutritive or aromatic constituents -from the original source material, and (in line with Frame Directive 89/107 /EEC) intentionally added to foods for the purpose of their colouration. [13]

Global harmonisation of food colours

The global trade requires harmonisation of food regulations on a world-wide basis in order to abolish barriers of trade and to ensure that the economical and nutritional demands of all nations are considered.
JECFA develops international standards for food additives considering toxicology and elaborating purity criteria. These standards are the basis of the standard for food colours of the General Standard for Food Additives from the Codex Alimentarius. It is not legally binding, but influences food colour regulations all over the world.

Colour EC No Natural occurrence Pigment
Re/blue E163 Black grapes, blackcurrants, cherries Anthocyanins
    elderberries, red cabbage, strawberries  
Pink E162 Beetroot Betanin
Red E120 Cochineal Carminik acid
Green E140 Alfafa grass, nettles, parley, spinach Chlorophylls
  E141   Chlorophyllins
  E160a (i) Annatto Mixed carotenes
  E160a (ii) Carrots Beta-carotene
Yellow E160b Oranges Bixin, norbixin
Orange E160c Prawns Capsanthin, Capsorubin
Red E160d Red pepper Lycopene
  E160e Tomatoes Apocarotenal
  E160f Palm fruit Apocarotenal (ethyl ester)
  E160b   Lutein
  E161g   Canthaxanthin
Yellow E100 Turmeric Curcumin
Yellow E101 Eggs, milk, yeast  
Black E153 Carbonised vegetable material Carbon black
Brown E150a-d Melanoidins (caramel) Melanoidins

B-ficoeritrina natural red colour

Looking for alternatives to artificial food colours such as sunset yellow, tartrazine and quinoline yellow Bermejo Ruperto, J. M. Alvarez-Pez and colleagues studied the marine algae Porphyridium cruentum and its protein B-ficoeritrina.

Phycoerythrin is a pigment of red algae and cyanobacteria. B-phycoerythrin and R-phycocyanin in native state, were obtained by the authors using an inexpensive and simple process from the red alga Porphyridium cruentum.

Removable edible paint for candies and dietary supplements

[16] A new form of coloured coating is being suggested by D. Tyler McQuade and colleagues. The coating is based on coloured calcium alginate hydrogels being useful in food such as candies and dietary supplements, cosmetic, medicinal, and textile uses and to wherever nontoxic, easily removable coloured coating is desired.

Tests were made on artificial turf spraying aqueous solutions of gelling agents: One layer of calcium chloride solution was followed by spraying sodium alginate solution with one per cent red food colour. The sprayed solutions then combined into a coloured thin film.

Gels with different physical properties were obtained varying the concentrations of the gelling agent. Low concentration of 0.1 moles/l of calcium chloride resulted in a brittle, easily removable film. High concentration of 2.0 moles/l of gelling agent resulted in a film which adhered well and was difficult to remove.

Flavours,flavour enhancer

These ingredients give taste,hide off flavour and permit standardisation of the taste of food.
They increase the value of the food bearing however the danger of excessive consume.


According to labelling rules of the European Community aromes can be included in the label without mentioning their origin. Artificial aromes and synthetic aromes are found under this class.Natural aromes are more valuable as artificial ones.They are more complex having therefore a better taste.The composition of synthetic flavours is more simple in their composition as natural ones. The taste is therefore not so specific.
Natural aromes are extracts of of spices such as vanilla and orange peeling.
Aroma, identical with natural aromas are synthetic origin.They have identical chemical structure of natural aromas.
Artificial aromas are of synthetical origin.They are not found in nature, for example: ethyl vanillin, methyl cumarine,resorcine dimethyl ether.

Smoke flavourings

Smoke flavourings: Only two out of 11 are safe, says EFSA

Nine smoke flavourings used in food production,were found not safe by the European Food Safety Authority (EFSA). Safety concerns, and possible risk of cancer could not be ruled out for one of those smoke flavourings. Only two were found to be safe.

All these flavourings are currently, or have previously been, on the market in the EU. They are added to meat, fish, cheeses, soups, sauces, drinks and confectionery to give them a "smoked" flavour, as an alternative to traditional smoking.

Of concern were TRADISMOKE A MAX, Scansmoke R909, Scansmoke PB 1110, SmokEz C-10, SmokEz Enviro 23, SmokEz Enviro 23, Zesti Smoke Code 10, AM 01, and Fumokomp which presented insufficient data. Of no concern were Scansmoke SEF7525 and Smoke Concentrate 809045. See the whole list [18] of assessed smoke flavourings and safety concerns at:

All smoke flavourings are being assessed according to EU Regulation 2065/2003 to see if they are suitable for human consumption. They will only be allowed for use in food if they are shown to be safe and are not a risk to health.

The Regulation 2065/2003EC states: " Because smoke flavourings are produced from smoke which is subjected to fractionation and purification processes, the use of smoke flavourings is generally considered to be of less health concern than the traditional smoking process.

Several international regulations cover smoke flavourings because of the concern about PAHs (Polycyclic Aromatic Hydrocarbons).

Summary of the Regulation EC 2065/2003 [19] The chemical composition of smoke depends among other things on the type of wood used, the method used for developing smoke, the water content of the wood and the temperature and oxygen concentration during smoke generation. Smoked foods in general give rise to health concerns, especially with respect to the presence of polycyclic aromatic hydrocarbons.

The production of smoke flavourings starts with the condensation of smoke. The condensed smoke is normally separated by physical processes into a water-based primary smoke condensate, a water-insoluble high-density tar phase and a water-insoluble oily phase. The water-insoluble oily phase is a by-product and unsuitable for the production of smoke flavourings.

The primary smoke condensates and fractions of the water-insoluble high density tar phase, the "primary tar fractions", are purified to remove components of smoke which are most harmful to human health. They may then be suitable for use as smoke flavouring.

Smoke is generated from wood which has not been treated with chemical substances during six month preceding felling. Herbs, spices, wigs of juniper and twigs, needles and cones of picea may be added if they are free of residues or chemical treatment. The source material is subjected to controlled burning, dry distillation or treatment with superheated steam in a controlled oxygen environment with a maximum temperature of 600°.

The smoke is condensed. Water and/or solvents may be added to achieve phase separation. Physical processes may be used for isolation, fractionation and/or purification to obtain the following phases:

Water-based "primary smoke condensate"

It contains mainly carboxylic acids, carbonylic and phenolic compounds, having a maximum content of:

benzo(a)pyrene 10µg/kg
benz(a)antracene 20µg/kg

"Water-insoluble high-density tar phase

This fraction precipitates during the separation phase and cannot be used as such for the production of smoke flavourings but only after appropriate physical processing to obtain fractions from this water-insoluble tar phase which are low in polycyclic aromatic hydrocarbons, already defined as "primary tar fraction" having a maximum content of:

benzo(a)pyrene 10µg/kg
benz(a)antracene 20µg/kg

"Water-insoluble oily phase"

If no phase separation has occurred during or after the condensation, the smoke condensate obtained must be regarded as a water-insoluble high-density tar phase, and must be processed by appropriate physical processing to obtain primary tar fractions which stay within the specified limits.

Contamination of foods with PAHs can happen by environmental PAHs that are present in air (by deposition), soil (by transfer) or water (by deposition or transfer), and during processing and cooking. The major contributors to PAH intake in the average diet are oils and fats, cereals, fruits and vegetables.

The waxy surface of vegetables and fruits can concentrate low molecular mass PAH through surface adsorption and particle-bound high-molecular-mass PAH can contaminate the surface due to atmospheric fallout.

Flavour enhancer

Flavour enhancer intensify flavour of food.Persons which are sensible to glutaminic acid may experience the "Chinese restaurant syndrome".There were related pressure on brainsides, headache,stiffness of neck[20].

On rats flavour enhancer cause alterations of reproduction and retarded learning.
Flavour enhancer potentiate voracity.
Flavour enhancer in human metabolism are transformed in uric acid which is undesired. Animals like rats do not form uric acid from flavour enhancers because they metabolise them as alantoine. Toxicological tests on rats are therefore irrelevant. Glutaminic acid is part of the proteins of our body. However there is a capital difference between glutaminic acid bound in a sequence of proteins and glutaminic acid or their salts being obtained synthetically. Glutaminic acid of the protein sequence is not free.

It is liberated during digestion and reaches the bloodstream slowly. Flavour enhancer are already free and reach the bloodstream immediately in great amount and may cause the above mentioned syndrome.[21]

People with the characteristic symptom should ask for food without glutamate.
Industry should reduce glutamate in their formulas and try to avoid completely its use in dry soups,dry sauces and an infinity of other product which are on market. Industry should return to natural ingredients avoiding synthetic other products. In doing so there is also a benefit on marketing because the products from natural resource have great acceptance by consumers. Please read the list of ingredients on the label, carefully, especially those of dry soups and dried sauces because they have a great amount of salts of glutamic acid.

The INTERMAP study links glutamate to obesity

Consumption of the flavour enhancer monosodium glutamate (MSG) may increase the risk of gaining weight, regardless of energy intake according to a study on humans by Ka He and colleagues 2008.

The authors cite that animal studies indicate that monosodium glutamate (MSG) can induce hypothalamic lesions and leptin resistance, possibly influencing energy balance, leading to overweight.

In his study He found that people with an average intake of 0.33 g/day of MSG in food preparation had an average BMI 23.5 kg per sq. m. and Non-MSG users had an average BMI of 22.3 kg per sq. m. The authors concluded that prevalence of overweight was significantly higher in MSG users than in non-users.

Reaction of the Glutamate industry

The Glutamate Association questions the study in a statement from 22.08.2008. The Association says that according to data of the WHO the countries with high intakes of glutamate do not have high population BMI.

The average person in the United States consumes approximately 11 grams of glutamate daily from all food sources (primarily dietary protein), while the body produces about 50 grams of free glutamate daily. Dietary glutamate from MSG averages less than one half gram/person/day .

Human studies where MSG was added to the diet have failed to show changes in body weight (Essed et al., Appetite 2007,48:29) [24]

A study on rats suggests that MSG in the diet actually suppresses body weight. (Kondoh and Torii, PhysiolBehav2008,doi:10.1016/j.physbeh.2008.05.010) [25]

E620 Glutamic acid

Glutamic acid occurs naturally in many foods and contributes to their flavour. Glutamic acid was firs obtained from gluten which gave the name to this aminoacid.

The flavour of its salt as monosodium glutamate was termed by professor Ikeda as umami.

E621 Sodium glutamate

Glutamate has a neurotransmitter function in the physiology of nervous cells.[26]
The neural function was used to promote selling of Intelligence Drugs to improve marks at school. Unfortunately there was no such benefit found.

Monosodium glutamate is one of the most abundant naturally occurring non-essential amino acids. Monosodium glutamate is used to improve the flavour and balances blends and rounds the total perception of other tastes in meals with meat, fish, poultry, many vegetables, sauces, soups, and marinades. It is, however not indicated in combination with sucrose. It improves the pleasantness only in the right concentration and becomes unpleasant in concentrations more than 1 gram per 100 ml in soups.

Chinese Restaurant Syndrome:The "MSG symptom complex" was originally termed the "Chinese Restaurant Syndrome" MSG became the focus and the symptoms have been associated with MSG ever since. In normal conditions, humans have the ability to metabolize glutamate that has a very low acute toxicity. The oral lethal dose to 50% of subjects (LD50) is between 15 to 18 g/kg body weight in rats and mice respectively, five times greater than the LD50 of salt (3 g/kg in rats). Therefore, the intake of MSG as a food additive and the natural level of glutamic acid in foods do not represent a toxicological concern in humans. [27]

E622 Potassium glutamate

E623 Calcium glutamate

E624 Monoammonium glutamate

E625 Magnesium glutamate

E626 Guanylic acid

E627 Sodium guanylate

E628 Potassium guanylate

E629 Calcium guanylate

E630 Inosinic acid

E631 Disodium inosinate

E632 Dipotassium inosinate

E633 Calcium inosinate

E634 Calcium 5'ribonucleotid

E635 Disodium 5'ribonucleotid

E640 Glycine and salts


It is used as flavour enhancer in sweet product mainly together with artificial sweetener s.It is used in chocolate, cakes and desserts.There is doubt about ethylmaltol being responsible for talasemia (a rare anaemia).

E239 Hexamethylentetramin

It is a widely used substance. as medicine against gout and infections of the urinary tract.It is also a vulcanisation accelerator and is used in the chemistry of explosives.In food it is a donator of formaldehyd and is used to to improve the optical appearance of food.At the moment it is used only in some kind of cheese.


It is an antibiotic used in infections of mouth,foot and genitals.It is employed in food industry to treat the shell of cheese. Resistance against this antibiotic will soon be established in bacteria coming in contact with it.His use should therefore forbidden in food industry


Antioxidants are used to improve the shelf life of food interfering in the reaction of oxygen with different components of food avoiding their chemical decomposition. They are used in soups,sauces in powder,chewing gum,dried products of potatoes margarineoil, icecream.

Measuring the antioxidant activity of phytochemicals

The methods used to evaluate antioxidant activity of phytochemicals such as food extracts, dietary supplements, fruits and juices are:

Oxygen radical absorbance capacity (ORAC)

ORAC is a method of quantitating the oxygen-radical absorbing capacity (ORAC) of antioxidants in serum using a few microliter. The ORAC assay determines the total antioxidant capacity of a sample which is estimated by taking the oxidation reaction to completion when all of the nonprotein antioxidants (which include alpha-tocopherol, vitamin C, beta-carotene, uric acid, and bilirubin) and most of the albumin in the sample are oxidized by the peroxyl radical. Results are quantified by measuring the protection produced by antioxidants. [28]

When comparing ORAC data, care must be taken to ensure that the units and food being compared are similar. Some evaluations will compare ORAC units per grams dry weight, others will evaluate ORAC units wet weight and still others will look at ORAC units/serving. [29]

Marketing uses ORAC as a selling argument for concentrated supplements claiming to be the number one ORAC product. However, there are no published the scientific literature so are difficult to evaluate on these values. [29]

Total radical-trapping antioxidant parameter (TRAP)

According to Bortolotti and colleagues total radical-trapping antioxidant parameter (TRAP) is being proposed to measure antioxidant property of plasma of diabetes patients. It may be either directly measured by a fluorescence-based method (TRAPm) or calculated (TRAPc) by a mathematicalformula, taking into account the serum levels of protein-bound SH (thiol) groups, uric acid, vitamin E, and vitamin C.

The authors found decreased TRAP levels in patients suggesting that antioxidant defenses in diabetes are lower than normal. They conclude that TRAP is more reliable than the measurement of each known antioxidants. In their research the authors found a correlation between TRAPc and TRAPc values, and suggest TRAPc, for routine assessment of oxidative stress in diabetic patients.

Trolox equivalent antioxidant capacity (TEAC)

The TEAC assay is based on the suppression of the absorbance of radical cations of 2,2'-azinobis(3-ethylbenzothiazoline 6-sulfonate) (ABTS) by antioxidants in the test sample when ABTS incubates with a peroxidase (metmyoglobin) and H2O2. If the inhibition time is fixed at 3 min.

There is lack of correlation between TEAC and other assays. This may result from underestimation of overall antioxidant capacity. Underestimation may be related to the effects of dilution and to premature measurement of inhibition percentage at a fixed time of 3 minutes.

Wang and colleagues conclude that the length of the inhibition time for the TEAC assay must be taken into account when determining the total antioxidant capacity of plasma, and that 30 min of inhibition is required for complete suppression of ABTS radical formation in the TEAC assay. The results from the TEAC assay at 30 min were similar and correlated with the results obtained by the ORAC assay over 70 min.

Total oxyradical scavenging capacity (TOSC)

The total oxyradical scavenging capacity (TOSC) is based on the oxidation of alfa-keto-y-methiolbutyric acid (KMBA) by 2,2'-azobis-amidinopropane (ABAP) with the evolution of ethylene as the quantifiable end product.

MacLean and colleagues modified the development of the assay of Winston et al. (Free Radical Biol. Med. 24 (1998) 480) and incorporated a standard curve resulting in a simple but reliable method to quantify the total water-soluble antioxidant capacity (TAC) of plant tissues such as apple fruit.

Peroxyl radical scavenging capacity (PSC)

Rui Hai Liu and colleagues developed an assay for assessing peroxyl radical scavenging capacity (PSC) of both hydrophilic and lipophilic antioxidant compounds and food extracts. It is based on the inhibition of dichlorofluorescin oxidation by antioxidants that scavenge peroxyl radicals, generated from thermal degradation of 2,2'-azobis(amidinopropane).

According to the authors the PSC assay is used to analyse or screen both hydrophilic and lipophilic antioxidants or food extracts and will be a valuable alternative biomarker for future epidemiological studies of chronic diseases.

Ferric reducing/antioxidant power (FRAP)

The ferric reducing antioxidant power (FRAP) assay determines directly the reducing capacity of a compound. Firuzi and colleagues used FRAP to study the antioxidant activities of 18 structurally different flavonoids. In this study it was found that the o-dihydroxy structure in the B ring and the 3-hydroxy group and 2,3-double bond in the C ring give the highest contribution to the antioxidant activity. [34]

Woodrow and colleagues used the FRAP assay, standardized against plant flavonoids and ascorbic aci, to quantify total antioxidant power in fruit samples, such as currants (Ribes nigrum), gooseberries (Ribes grossularia), raspberries and blackberries (Rubus spp.). Black currant demonstrated the highest antioxidant activities followed by raspberries and gooseberries. Furthermore, the authors found that some wild Rubus species exhibited activities significantly higher than the cultivars. [35]

DPPH free radical method

Zheng DeYong and An XinNan presented a method for determining 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging activity of natural antioxidants. The DPPH free radical scavenging activity of 21 species of cluster-bamboo's leaf were determined.

New cellular antioxidant activity (CAA) assay

Kelly Wolf and colleague developed the cellular antioxidant activity (CAA) assay using the bioactivity inside human hepatocarcinoma HEPG2 cells, which relies on cellular biological activity rather than chemicalreactions which may not mirror reactions inside a living cell.

Dichlorofluorescin trapped within HEPG2 cells is subjected to oxidation by 2,2'-azobis(2-amidinopropane) dihydrochloride (ABAP)-generated peroxyl radicals. During the oxidation fluorescent dichlorofluorescein (DCF) and cellular fluorescence is produced. The difference between the fluorescence of control cells and those which with added solution of the foods on test indicates the antioxidant capacity of the compounds. Results of CAA assay are expressed in micromoles of quercetin equivalents per 100 micromol of phytochemical or micromoles of quercetin equivalents per 100 g of fresh fruit.

The CAA values from phytochemicals decreased from quercentin, kaempferol, epigallocatechin gallate (EGCG myricetin, luteolin, gallic acid, ascorbic acid, caffeic acid, to catechin.

The CAA values decreased from Blueberries, craberry, apples, red grapes to green grapes. The authors conclude that the CAA assay is more biologically relevant than known chemical reaction because it considers uptake, metabolism, distribution and location of antioxidant compounds within cells.

Classes of Antioxidants

There are two main antioxidants classes:

Vitamins as dietary supplements

There are many dietary supplements being sold at supermarkets.The prices differ enormously.The amount of vitamins are almost the same and cannot justify the difference of price between the products.

Some products have minerals and other trace elements varying from product to product.Their benefit should be analysed considering the local nutritional habits.
It is is very difficult to establish general rules concerning the amount of daily supplements to be taken because of different nutritional habits. To explain these problems the case of selenium, potassium, magnesium and vitamin A is here cited:

Brazil nuts are a good source of selenium

Christine D Thomson and colleagues 2008 assessed the bioavailability of selenium from Brazil nut, compared with seleniumethionine used to fortify foods and in dietary supplements. They found that the consumption of 2 Brazil nuts daily is as effective for increasing selenium status and enhancing GPx activity as 100 µg Se as selenomethionine. The authors suggest to include Brazil nut in the diet to avoid to fortify foods or use supplementation of selenium in New Zealand.

This would also benefit residents in Europe where the recommended daily intake (RDI) is 65 micrograms. The selenium intake in UK is estimated to have fallen from 60 to 34 micrograms. on account of reduced import of selenium rich wheat from U.S.

Radioactivity of Brazil nuts

However, an unhampered consume of Brazil nuts should consider the high radiactivity of the nut. According the UK DEFRA, among other foodstuffs, Brazil nuts contain some of the highest levels of natural radioactivity, in particular radium-226 and radium-228. The consumption of a 100g bag (about 30-40 nuts) per week (about 5 nuts a day) throughout the year would give rise to an annual dose of 0.2 mSv.

Ingestion and inhalation of long-lived natural radionuclides are estimated to give rise to an average annual dose of 0.27 mSv. Potassium and magnesium stabilises heart rhythmic disturbaces. Both minerals should be supplemented up to 50% from the normal daily intake of 2-3 grams for potassium and 200-350 milligramme for magnesium. People with this kind of disorder need therefore supplementation of these minerals. Normal people can supply the minerals out of a balanced nutrition. [41] Vitamin A is important for the north and the south of the globe because of having less sun incidence. People living near the equator don't have to bother supplementing with vitamin A. Because of high incidence of sunlight boosting biological synthesis of vitamin A of the body in these countries one fears an hypervitaminosis of vitamin A. Therefore regulations in these countries forbid adding vitamin A to food.

Safety and bioavailability of Vanadium, EFSA and FSA assessment

[42] [43]
EFSA's AFC Panel has issued an unfavourable opinion on vanadium containing compounds which may be added for nutritional purposes in foods for particularly uses and foods (including food supplements) intended for the general population. The assessed compounds were vanadium citrate, bismaltolato oxo vanadium and bisglycinato oxo vanadiumand vanadyl sulphate, vanadium pentoxide and ammonium vanadate. The non-vanadium constituents of these sources of vanadium are of no safety concern at the levels considered in this opinion. However, according to the FSA NDA Panel vanadium itself present various toxic effects, and a tolerable upper intake level could not be established in lack of appropriate data.

The bioavailability of vanadium from five of these six compounds is higher than that of vanadium absorbed from the normal diet. Consequently consumers could be exposed to higher levels of vanadium through products containing these five compounds than from a normal diet.

The Panel concluded that the safe use of the six sources for vanadium added to foods intended for the general population, including food supplements, and foods for particular nutritional uses, could not be established.

Uses of vanadium

These sources for vanadium are not included in the list of vitamin and mineral substances which may be used in certain foods including food supplements. However they have been allowed to remain in use in Member States of the European Union.
Vanadium forms are typically used as a dye and colour-fixer in foods and supplements. Vanadyl sulfate, has been used to increase insulin sensitivity in supplements and has therefore been targeted at diabetes sufferers and the body-building market.

Occurrence of vanadium in food, food supplements

According to the UK Food Standards AgencyBeverages, fats, oils, fresh fruits and vegetables contain the lowest levels of vanadium, whereas whole grains, seafood, meats and dairy products contain more (0.005 - 0.03 mg/kg). A few foods, including spinach, parsley, mushrooms and oysters, contain relatively high amounts of vanadium (>0.10 mg/kg).

Vanadium is present in a number of multi-vitamin/mineral dietary supplements at levels of approximately 0.025 mg per day. There are no licensed medicines containing vanadium.

Other sources of exposure

Exposure to vanadium by inhalation may occur occupationally. In the production of vanadium pentoxide, dust concentrations of the pentoxide can range from 0.1 to 30 mg/m³ , and concentrations of 0.5-5 mg/m³ are not uncommon in the production of vanadium metal and vanadium catalysts.

Recommended amounts

Vanadium has not yet been proven to be an essential trace element for mammals. There is no evidence to suggest that the vanadium we get from food is harmful. It is unlikely that we need vanadium for good health and too much could be harmful.

Function of vanadium

No specific function has been identified for vanadium in higher animals. In vitro and animal studies suggest that vanadium may function as an oxidation-reduction catalyst, and may regulate the sodium, potassiumadenosine triphosphatase enzyme, however, this has not been proven.

Deficiency of vanadium

In humans, the reported signs of deficiency are questionable, although it has been suggested that low intakes may be associated with cardiovascular disease.


Although no specific data have been identified, it is possible that vanadium may interfere with the storage and metabolism of iron, because absorbed vanadium is bound to transferrin.

Absorption and bioavailability

Intestinal absorption of vanadium is low, less than 5%. The mechanism of absorption has not been defined.


Absorbed vanadium is mainly transported in the plasma, associated with transferrin. Concentrations reported in human blood vary widely, with levels in whole blood and serum in the range of 0.01 - 0.4 mg/L.

The concentrations in all tissues are low, but are higher in the liver, kidney and lung. Vanadium is also present in breast milk and saliva and passes through the blood brain barrier. Small amounts have been identified in the placenta. Based on animal studies, bones and teeth retain the highest concentrations of vanadium.


Ingested vanadium is predominantly eliminated unabsorbed via the faeces. Absorbed vanadium is mainly excreted via the urine.

Toxicity, Human data

The toxicity of vanadium compounds increases as valency increases, V5+ being the most toxic. In humans, exposure by inhalation causes diverse toxic effects on the respiratory, digestive, and central nervous systems, the kidney and skin. There are very few reported cases of vanadium toxicity in humans, when it is taken by mouth.

Supplementation trials

Supplementation of human volunteers with vanadyl compounds at oral doses of 50-125 mg/day caused cramps, loosened stools and "green tongue" in all patients, and fatigue and lethargy in a minority.

Animal data

Orally administered vanadium has low overt toxicity, but is reported to have adverse effects on reproduction and development in both males and females. There is some evidence of increased pre and post-implantation foetal loss and significant accumulation of vanadium in the foetus. Skeletal anomalies and reduced ossification in the offspring, as well as an increased incidence of cleft palate have been reported.

Carcinogenicity and genotoxicity

Lifetime studies in animals indicate that vanadium is not carcinogenic. Positive results have been obtained in some in vitro mutagenicity tests.

Exposure assessment of vanadium, Total exposure/intake

Food Mean: 0.013 mg/day (1980 UK TDS)
Supplements up to 0.025 mg/day (Annex 4)
Drinking Water 0.01 mg/day (estimated from 0.005 mg/L, WHO 1988)
Estimated maximum daily intake 0.013 + 0.025 + 0.01 = 0.05 mg/day The table below gives some information about the usual products on the German market:

Ingredients Centrum Multibionta Multivitamin Krüger Hermes Multivit
Vitamin A 800µg - - -
Provitamin A - - - 2 mg
Vitamin E 10 mg 12 mg 10 mg 12 mg
Vitamin C 60 mg 300 mg 60 mg 75 mg
Vitamin K1 30µg - - -
Vitamin B1 1,4 mg 1,3 mg 1,4 mg 1,6 mg
Vitamin B2 1,6 mg 1,7 mg 1,6 mg 1,8 mg
Vitamin B6 2,0 mg 1,8 mg 2,0 mg 2,1 mg
Vitamin B12 1 µg 3 µg 1 3 µg
Vitamin D 5 µg - - -
Biotin 150 µg 30 µg 0,15 mg 30 µg
Folic acid 200 µg 150 µg 200 µg 160 µg
Nicotinamid - 18 mg 18 mg -
Niacin 18 mg - - 20 mg
Pantothenate 6 mg 8 mg 6,0 mg 6,5 mg
Calcium 162 mg - - 100 mg
Phosphate 125 mg - - -
Iron 4 mg - - -
Magnesium 100 mg - - -
Iodine 100 µg - - -
Potassium 40 mg - - -
Chloride 36,3 mg - - -
Copper 1 mg - - -
Manganese 1 mg - - -
Chromium 25 µg - - -
Molybdenum 25 µg - - -
Selenium 25 µg - - 10 µg
Silicium 2 µg - - -
Zinc - - - 2 mg

Food supplements should have a ratio of half as much magnesium as calcium. A daily intake of 1.200 mg calcium needs 600 mg of magnesium[45].

Ice structuring proteins

Ice-structuring proteins (ISPs) in edible plants and fish that need to protect themselves against freeze damage. ISPs have potential applications in a number of areas including cryopreservation and frozen foods manufacture.

ISP type III HPLC 12 being of particular interest. No evidence of a genotoxic potential or notable subchronic toxicity were found by T. Hall-Manning during a safety evaluation of ice-structuring protein (ISP) type III HPLC 12.[47]

Anti-freeze protein, also called "ice structuring proteins" (ISPs) found in Ocean Pout (Marcrozoarces americanus) has been approved for use as ice structuring proteins in ice cream improving flavour and texture of the product using less sugar and fat. It is obtained from the fermentation of a genetically modified food grade yeast (Saccharomyces cerevisiae). This affects a huge market of dairy ice cream, milk ice, water ice, fruit ice, sorbets, frozen deserts and any similar products.

Professor Malcolm Hooper from Sunderland University, Professor Joe Cummins from the University of Western Ontario in Canada, and Dr Mae-Wan Ho, director of the Institute of Science in Society are against EU approval of ISP alleging that it could cause serious allergies. They say Unilever tests with people allergic to cod, not to ocean pout were insignificant.

AFP claims that their antifreeze proteins type III are all purified from their natural sources cold ocean teleost fish.

EFSA considers ice structuring protein ISP as safe

Ice structuring proteins (ISPs) are naturally produced by a variety of living organisms - including certain fish, plants and vegetables - to help them cope with very cold environments by lowering the temperature at which ice crystals form.

A joint opinion on the EFSA found these proteins to be safe for use in foods. They will be added to ice cream in order to control the formation of ice crystals during manufacture permitting a creamy consistency with lower fat content. The technique involves production of the isolated proteins using a genetically modified strain of baker's yeast. The protein produced does not contain any residual modified yeast cells or detectable recombinant DNA.

EFSA's NDA and GMO Panels concluded that the proposed use of ISPs - in ice cream at no more than 0.01% of weight - is safe subject to the specification and production practices described by the applicant. The Panels found no evidence of genotoxic activity in a variety of trials. Based on a range of test results, the risk of an allergic reaction in fish-allergic people or the population at large is considered very unlikely, as is the possibility that allergic reactions to yeast allergens could occur due to eating the ISP-containing products.

Ice structuring proteins are in common foods such as oats, rye, wheat, barley, carrot, potato and cold water fish. No safety issues have been reported either from consuming natural dietary sources or through the addition of ISPs to foods, which is authorized in countries including the United States, Australia and New Zealand.

The organism used for the production of ISPs

The baker's yeast Saccharomyces cerevisiae. Strain CENPK338 has been used since 2003 for commercial production of the ISP for use in non European ice cream production. No gene encoding antibiotic resistance and no bacterial DNA were introduced. From 2003 to 2007 more than 470 million ISP-containing edible ice products have been sold in the USA and 47 thousand litres of ISP containing ice cream has been sold in Australia/New Zealand. There have been no reported safety issues.

With regards to the potential of adverse allergic reactions against yeast allergens, the Panel considers it is unlikely that such reactions would occur after ingestion of the ISP-containing products. The Panel concludes that the use of the ISP type III HPLC 12 preparation at a maximum level equivalent to 0.01% ISP type III HPLC 12 in edible ices is safe.


[49] Enzymes play an increasingly important role in food production, and can be used as alternatives to chemicals in improving the texture, appearance, nutritional value and flavour of food, as well as helping in certain food production processes (e.g. helping bread to rise). Currently food enzymes used as processing aids are not covered by EU legislation. Member States' legislation on food enzymes differs significantly, which can lead to problems for the internal market and an unclear situation for the EU consumer.

Historically, food enzymes were considered to be non-toxic. However, the food enzyme industry is continually striving to develop improved technology resulting in the development of food enzymes which became through the years more complex and sophisticated. There could be some potential hazards arising from their chemical nature and source such as allergenicity, activity-related toxicity, residual microbiological activity, and chemical toxicity. Therefore safety evaluation of all food enzymes, including those produced by genetically modified micro-organisms (GMOs), is essential in order to ensure consumer safety.

Existing provisions in the area of enzymes

Directive 95/2/EC on food additives other than colours and sweeteners allows for the use of two enzymes as food additives: E1103 Invertase and E1105 Lysozyme.In addition, Council Directive 2001/112/EC relating to fruit juices and certain similar products intended for human consumption, Council Directive 83/417/EEC relating to certain lactoproteins (caseins and caseinates) intended for human consumption and Council Regulation (EC) No 1493/1999 on the common organisation of the market in wine, regulate the use of certain food enzymes in these specific foods.
Under the new proposed legislation, harmonised EU rules would be laid down for the evaluation, approval and control of enzymes used in food. The draft Regulation foresees the way to draw up an initial positive list of enzymes.

The proposal also includes requirements for the labelling of food enzymes other than those used as processing aids. Food enzymes with a technological function in the final food will have to be labelled as ingredients with their function (e.g. stabiliser) and name.

Labelling of enzymes

In most cases food enzymes will be used as processing aids i.e. will be present in food in the form of a residue, if at all and will have no technological effect on the finished product. Taking into account that all food enzymes will be assessed for their safety, it is proposed that food enzymes which are used as processing aids are exempted from labelling.

Food enzymes used to exert a technological function in the final food, will be labelled with their function (e.g. stabiliser etc) and specific name.

Ezyme Commission number (EC number)

The Enzyme Commission number (EC number) is a numerical classification scheme for enzymes, based on the chemical reactions they catalyze. As a system of enzyme nomenclature, every EC number is associated with a recommended name for the respective enzyme.

Every enzyme code consists of the letters "EC" followed by four numbers separated by periods. Those numbers represent a progressively finer classification of the enzyme. For example, the enzyme tripeptide aminopeptidase has the code "EC", whose components indicate the following groups of enzymes: EC 3 enzymes are hydrolases (enzymes that use water to break up some other molecule), EC 3.4 are hydrolases that act on peptide bonds, EC 3.4.11 enzymes are only those hydrolases that cleave off the amino-terminal amino acid from a polypeptide, and EC are those that cleave off the amino-terminal end from a tripeptide.

Strictly speaking, EC numbers do not specify enzymes, but enzyme-catalyzed reactions. If different enzymes (for instance from different organisms) catalyze the same reaction, then they receive the same EC number.

Group Reaction catalyzed Typical Trivial
    reactions names
EC 1 To catalyse oxidation/reduction AH + B -A+ Dehydrogenase,
Oxyreductases reactions; transfer of H and O atoms BH (reduced) oxidase
  or electrons from one substance A + O - AO  
  to another (oxidized)  
EC 2 Transfer of a functional group from one AB + C - A + Transaminase,
Transferases substance to another. The group may BC Kinase
  be methyl-, acyl-, amino- or phospate    
EC 3 Formation of two products from a AB + H2O - Lipase, amylase,
Hydrolases substrate by hydrolyses AOH + BH peptidase
EC 4 Non-hydrolytic addition or removal of RCOCOOH -  
Lyases groups from subnstrates. C-C, C-N, C-O RCOH + CO2  
  or C-S bonds may be cleaved    
EC 5 Intramolecule rearrangement, i.e. AB - BA Isomerase,
Isomerases isomerization changes within a single   mutase
EC 6 Join together two molecules by X + Y + ATP -> Synthetase
Ligases sythesis of new C-O, C-S, C-N or C-C XY + ADP + Pi  
  bonds withg simultaneous breakdown of    


UniProt is the universal protein database, a central repository of protein data created by combining Swiss-Prot, TrEMBL and PIR. This makes it the world's most comprehensive resource on protein information. The UniProt Consortium is comprised of the European Bioinformatics Institute (EBI), the Swiss Institute of Bioinformatics (SIB), and the Protein Information Resource (PIR). UniProt identifiers uniquely specify a protein by its amino acid sequence.

EBI, located at the Wellcome Trust Genome Campus in Hinxton, UK, hosts a large resource of bioinformatics databases and services.

SIB, located in Geneva, Switzerland, maintains the ExPASy (Exprt Protein Analysis System) servers that are a central resource for proteomics tools and databases.

PIR, hosted by the National Biomedical Researche Foundation (NBRF) at the Georgetown University Medical Center in Washington, DC, USA, is the oldest protein sequence database, Margaret Dayhoff's Atlas of Protein Sequence and Structure.

In 2002, EBI, SIB, and PIR joined forces as the UniProt Consortium.

Combination of food additives and interference with development of nervous cells

The effects of food colouring brilliant blue (E133) combined with monosodium glutamate (MSG; E621) and colouring quinoline yellow (E951) combined with the sweetener aspartame (E951) were tested at the University of Liverpool. This combination presented neurotoxic effects.

The combination of brilliant blue and MSG could inhibit cell growth up to four times more than the additives on their own, while for quinoline yellow and aspartame the figure rises to seven. The research has been supported by Organix Brands.

Inhibition of neurite outgrowth was found at concentrations of additives theoretically achievable in plasma by ingestion of processed foods like sweets and snacks they are typically present in combinations and are foods which are consumed by children whose nervous system is still developing. [52]

In response to the study, UK regulatory body Food Standards Agency said that the safety of all additives is kept under review. The European Food Safety Authority (EFSA) had been asked to review food additives currently permitted within the EU in order to determine whether full re-evaluation is required. [53]

The authors of the Liverpool study call upon the European Food Safety Authority (EFSA) to look upon the combined effects of the mentioned substances. Some researches concerning mixtures of substances are already being undertaken by FSA funding research on the effects of two groups of colour additives on the behaviour children is expected to report in 2007.

The European Commission protects European sodium glutamate producer

Monosodium glutamate (MSG) is a flavour enhancer for foods such as soups, fish and meat dishes, and ready meals in produces of Nestle, Unilever and others. Glutamate was linked to headache, known as the China restaurant syndrome and other problems.

Ajinomoto Foods Europe, the only MSG producer in the EU, complainted of Chinese firms dumping sodium glutamate on the EU market undercutting Ajinomoto's price by up to 24 per cent. Following the complaint, the European Commission imposed in 2008 anti-dumping tariffs of 39,7 per cent on monosodium glutamate (MSG) imported to the EU from China.

Retinal cell destruction caused by sodium glutamate

Ohguro Hirishi and colleagues 2002 found a significant accumulation of glutamate in vitreous was observed in rats on high sodium glutamate. Thickness of retinal neuronal layers was remarkably thinner in rats fed on sodium glutamate diets than in those on a regular diet.

The authors concluded that a diet high in sodium glutamate over a period of several years may increase glutamate concentrations in vitreous and may cause retinal cell destruction.

Monosodium Glutamate not related to the Chinese Restaurant Syndrome

Jinab and Hajeb 2010 of the University Putra Malaysia reviewed application, benefits of monosodium glutamate as flavor enhancer. The authors say that glutamate adds a fifth basic taste to the four basic ones, whicch are saltiness, sourness, sweetness and bitterness. It is also an energy source, acts as a substrate for glutathione synthesis and enhances food intake in older individuals. Glutamate may partially replace salt in food preparation. The Joint Expert Committee on Food Additives of the United Nations Food and Agriculture Organization and World Health Organization classified glutamate as safe. The authors stress that there are no consistent clinical data to support believes that glutamate causes asthma, migraine headache, Chinese Restaurant Syndrome, and there are no evidences indicating that individuals may be uniquely sensitive to glutamate.

The position of the Food Standards Australia New Zealand

The Food Standards Australia New Zealand in a technical report of 2003 found that some studies reported a complex of symptoms which came to be known as the Chinese restaurant syndrome (CRS) because they typically followed ingestion of a Chinese meal. Two outstanding studies were Kwok, R. (1968) [58] and Schaumburg HH(1969) [59] suggesting monosodium glutamate as the causative agent in CRS embracing symptoms such as headache, numbness/tingling, flushing, muscle tightness, and generalised weakness. MSG symptom complex is now being used instead of CRS. A possible association between MSG and bronchospasm in asthmatic individuals were also suggested.


The FSA study found no convincing evidence that MSG is a significant factor in causing systemic reactions resulting in severe illness or mortality, and studies have failed to demonstrate a causal association with MSG. Some reactions were noted by administrating large doses of 3g or more MSG without food were not serious and are likely to be attenuated when MSG is consumed with food. Bronchospasm in asthmatic individuals is, according to actual data, not significantly triggered by MSG.

Patricia Tagliaferro in an article of 1995 [60], stressed inconsistent data of studies on the possible effect ofMSG. The study of Jinab and Hajeb 2010 confirms that glutamate is not related to asthma, migraine headache and Chinese Restaurant Syndrome.

SodiumBenzoate and certain colours increases hyperactivity in children

According to the UK Food Standards Agency, parents of children showing signs of hyperactivity are being advised that cutting certain artificial colours from their diets might have some beneficial effects.

The colours found to give adverse reactions were: Sunset yellow (E110), Quinoline yellow (E104), Carmoisine (E122), Allura red (E129), Tartrazine (E102) Ponceau 4R (E124), and Sodium benzonate (E211)

Defining Hyperactivity

Hyperactivity is the occurrence of the following behaviours at the same time: over-activity, inattention and impulsivity. Attention Deficit Hyperactivity Disorder (or Hyperkinetic Disorder) (ADHD) is an extreme form of hyperactivity that is clinically diagnosed when specific patterns of behaviour occur together to a strong degree. This comprises a behavioural disorder which adversely affects children's function at home and in school.

The findings

The Committee on Toxicity of Chemicals in Food (COT) reviewed a research, carried out by Southampton University, suggesting that eating or drinking certain mixes of these artificial food colours together with the preservative sodium benzoate could be linked to a negative effect on children's behaviour.

The findings of the research were presented to the European Food Safety Authority (EFSA), which is conducting a review of the safety of all food colours.

FSA Advice

Following the COT statement, the FSA holds on the following advice:
FSA advice to consumers: if a child shows signs of hyperactivity or Attention Deficit Hyperactivity Disorder (ADHD) then eliminating the colours used in the Southampton study from their diet might have some beneficial effects.

However, the FSA also reminds that there are many factors associated with hyperactive behaviour in children. These are thought to include genetic factors, being born prematurely, or environment and upbringing.

The Southampton Study

[62] [63]
The study tested two combinations of colours and a preservative most likely to be found in foods popular with children such as soft drinks, confectionery, and ice cream.

According to Professor Jim Stevenson from Southampton University, and author of the report, the consumption of certain mixtures of artificial food colours and sodium benzoate preservative are associated with increases in hyperactive behaviour in children.

The POC Review on the Southampton Study

According to Professor Ieuan Hughes, Chair of the COT, said that the study provides supporting evidence for a link between the colours used in the study and increased hyperactivity in children. He stresses that the available evidence does not identify whether this association would be restricted to certain food additives or combinations of them.

The POC study concludes that the findings are consistent with, and add weight to, previous published reports of behavioural changes occurring in children following consumption of particular food additives.

German BFR agrees with UK opinion on Hyperactivity and Additives

A trial by Southampton University, commissioned by the British Food Standards Agency (FSA), examines a possible association between the intake of specific food additives (the food colourings E102, E104, E110, E122, E124, E129 and the preservative sodium benzoate E211) and the occurrence of Attention Deficit Hyperactivity Disorder (ADHD) in children. The trial found that artificial colours or a sodium benzoate preservative (or both) in the diet result in increased hyperactivity in 3-year-old and 8/9-year-old children in the general population. [66]

The COT Opinion

The UK Committee on Toxicity of Chemicals in Food, Consumer Products and the Environment (COT) considers the results of the trial by Southampton University as additional indications of a possible association between the intake of certain mixes of artificial colouring agents containing the preservative sodium benzoate and increased hyperactivity in children.

To the extent that there is a causal association, this could be of importance for individual children particularly for those who are in any case clearly hyperactive. However, COT stresses that the mean levels of observed hyperactivity are low compared to normal inter-individual variation and that behavioural changes did not occur in all children in one group, did not occur uniformly across all age groups and not in an even manner for the intake of all additive groups. Hence it is not possible to draw any more extensive conclusions. Nor is it possible to extrapolate the results to other additives.

The German BfR - Federal Institute for Risk Assessment opinion

After short-term examination of the publication by McCann et al. (2007) [66] BfR agrees with the conclusions in the COT opinion. As food additives must be included in the list of ingredients, consumers wishing to avoid any intake of the examined substances for precautionary reasons are able to refrain from consuming the corresponding foods and drinks.


The Southampton study, together with the review of COT, should encourage food producers to reformulate their products to reduce as much as possible the use of additives.

The Southampton Study on hyperactivity does not warrant an ADI change of colour and preservative, says EFSA.

The European Food Safety Authority's (EFSA) AFC Panel assessed the study of McCann and al 2007 on hyperactivity. The report was released on 14.03.08 concluding that the McCann study (2007) provided limited evidence that the mixtures of additives tested had a small effect on the activity and attention of some children. However, the effects observed were not consistent for the two age groups and for the two mixtures used in the study.

Considering the overall weight of evidence and in view of the considerable uncertainties the Panel concluded that the findings of the McCann et al study could not be used as a basis for altering the ADI of the respective food colours or sodium benzoate.

Although the findings from the study could be relevant for specific individuals showing sensitivity to food additives in general or to food colours in particular, it is not possible at present to assess how widespread such sensitivity may be in the general population.

The Panel noted that the majority of the previous studies used children described as hyperactive and these were therefore not representative of the general population.

Southampton team responds to EFSA evaluation

The researcher of the Southampton study on hyperactivity caused by certain colourings and preservative, leaded by Jim Stevenson, Donna McCann, Edmund Sonuga-Barke and John Warner, responded to the EFSA evaluation of their study. They say that despite EFSA saying there is no justification from this research to change the limits on these additives, that does not mean there are no grounds for action at all.

The team stresses that since the colours being tested in this study are of no nutritional value, even the small overall benefit of removing them from children's diets would come at no cost or risk to the child, and a benefit, even a small one, would be worthwhile achieving.

According to the authors added weight is given to this conclusion because other important influences on hyperactivity in children, such as genetic factors, are difficult to address while the risk arising from exposure to food colours can be regulated.

Food colours and preservative should be eliminated despite all-clear of EFSA. Attention-Deficit Hyperactivity Disorder (ADHD) and food additives

The Southampton study found that a mix of food colours such as sunset yellow (E110), tartrazine (E102), carmoisine (E122), ponceau 4R (E124) quinoline yellow (E104), allura red (E129) and sodium benzoate (E210). exacerbate hyperactive behaviour in children at least up to middle childhood.

EFSA review

The European Food Safety Authority (EFSA) conducted a review of the Southampton study, as well as other studies and meta-analyses on additives and hyperactivity. EFSA also concluded that the Southampton study gave no basis for changing acceptable daily intakes (ADI) of food additives. EFSA stresses that hyperactivity results from a variety of social and biological causes, focusing on dietary strategy eliminating food additives alone may detract from the provision of adequate treatment.

Professor Kemp position

At present, there are three main approaches to tackling ADHD: drugs, behavioural therapy, and dietary management.

Despite all controversies Professor Andrew Kemp writes that behavioural therapy is still considered necessary for adequate treatment even though there is less evidence for its efficacy than there is for eliminating food additives. He calls not to neglect the dietary factors. Cutting out colours and preservatives from the diets of hyperactive children should be standard part of the treatment of ADHD.

Aldi Australia banned six food colourings

Aldi Australia said it will no longer wait for the food regulator to act. The supermarket chain has voluntary banned six food colourings from its products following the results of a research published by The Lancet in 2007. The artificial colours sunset yellow (E110), tartrazine (E102), carmoisine (E122), ponceau 4R (E124), quinoline yellow (E104), and allura red (E129) have been linked to behavioural problems in children such as ADHD.

The Lancet Study

The British Southampton study published in the "Lancet" in 2007 found a link between hyperactivity in children who consumed drinks that contained these additives.

The study tested two combinations of colours and a preservative most likely to be found in foods popular with children such as soft drinks, confectionery, and ice cream.

According to Professor Jim Stevenson from Southampton University, and author of the report, the consumption of certain mixtures of artificial food colours and sodium benzoate preservative are associated with increases in hyperactive behaviour in children.

List of additives you should avoid

A list of food additives which should be avoided is given by Food Intolerance Network. Please see:

The European situation

The European Parliament voted in July 2008 for products containing the colours to be labelled "may have an adverse effect on activity and attention in children".

The European Food Safety Authority (EFSA) reviewing the methodology and results of the Southampton study stated it found no scientific evidence for altering intake recommendations of any of the additives, but will look again at the safety data on all food additives, including the Southampton colours. [74]

The power of marketing

Aldi Australia demonstrated a high level of quality and responsibility for the health of its customers. Aldi Europe and Lidl Europe still sell products with the Southampton colours. The marketing department should be aware of the possible image damage selling products which might harm children. Changing the six questioned artificial colours to other innocuous ones clears the situation and demonstrates a precautious way to handle health issues.

Aldi Europe and LIDL still sell products with the Sothhampton colours

Image Lollies

Paprika food colour is safe

Paprika as food colour is increasingly being used by the food industry. Safety tests on rats found paprika food colour as safe. The toxicity and carcinogenicity studies of paprika colour were performed by Inoue and colleagues 2008.

Treatment with paprika colour caused a significant increase in incidence of hepatocellular vacuolation, in 5% males, however, no toxicological effects or other abnormal effects such as tumors were noted.

The authors concluded that, based on slight histopathological changes, noted as hepatocellular vacuolation, the no-observed-effect level (NOEL) was estimated to be 2.5% in the diet (1253 mg/kg bw/day) and the the no-observed-effect level (NOEL) was 2.5 per cent in the diet, or 1253 mg per kg of bodyweight per day, for the male rats. The NOEL for the female rats was determine to be five per cent, or 2826 mg per kg of bodyweight per day.
Paprika colour is therefore not carcinogenic nor toxic under present conditions.

Connection between diet and behaviour in attention-deficit hyperactivity disorder (ADHD) children

[76] As part of the Impact of Nutrition on Children with ADHD (INCA) study Pelsser and colleagues found that a 5 week restricted elimination diet consisting mainly of hypoallergenic foods such as rice, turkey, lamb, a range of vegetables (lettuce, carrots, cauliflower, cabbage and beet), pears and water. The hipoallergic diet reduced the ADHS symptoms by at least 40% in 78% of the children, compared with a control group. Diets were composed of high-IgG or low-IgG foods, based on every child's individual IgG blood test results. Children which returned to high allergic diet had a relapse in symptoms with no difference in the immunologic responses immunological response to the different diets. The authors concluded that food sensitivity plays a part in ADHD, but it is not caused by an allergic reaction, and diets on the basis of IgG blood tests should be discouraged.

The same authors, in a foregoing study in 2010, report significant reductions of complaints of these children related to three domains: headaches or bellyaches, unusual thirst or unusual perspiration, and sleep complaints were significantly reduced by hypoallergenic diet, but these reductions were independent of the behavioural changes. An elimination diet may be an effective instrument to reduce aforementioned physical complaints in children with ADHD. [77]

Artificial colour in drinks must be reduced: Sunset Yellow E110 [78]

The EU Standing Committee on the Food Chain and Animal Health followed the suggestion of the European Food Safety Authority (EFSA) to reduce the the usage levels of Sunset Yellow for flavoured drinks from 50 mg/litre down to maximum 20 mg/Litre, to be valid by the end of the year.

Artificial colours are ruled by the Directive 94/36/EC [11]. The ADI for Sunset Yellow was reduced to 1 mg/kg/bw/day after noting that the exposure was too high in some population groups and was not safe at higher levels in food and drinks.

Suggested limit of 10 mg/l was strongly rejected by the drink industry as too low Soft drink producer heavily lobbied the First Minister Alex Salmond who wrote to the European Commissioner for Health and Consumer Policy John Dalli warning that reducing the additive level to 10 mg could have a negative impact on the "iconic" drink. So AB Barr, producer of the ditching yellow drink, succeeded to have a minimum of 20 mg/litre of Sunset Yellow for Irn Bru.
Safety of artificial colours are being questioned following the Southampton study in 2007. [62] [64]

Artificial colour E124 Ponceau 4R

To meet the regulations for food colouring of tThe U.S. Food and Drug Administration.prohibits the use of E124 Ponceau 4R (also called Cochenill Red A) used in the UK formulation. To meet US regulations Barr uses alternative food and drink colourants to market its soft drink in the US. [79]

Sugar, anti-social behaviour and ADHD

Sugar and adverse conduct problems

Lien et al 2006 found a relationship between soft drink consumption and mental distress, conduct problems, and total mentalhealth difficulties score. The higher the consumption of soft drinks, the more extreme symptoms of hyperactivity were observed. The highest adverse reaction observed for conduct problems were found among boys and girls who consumed 4 or more glasses of sugar-containing soft drinks per day.

The authors concluded that high consumption levels of sugar-containing soft drinks were associated with mental health problems among adolescents.

Sugar is not linked to anti-social behavior

David Benton, in a response to the study of Lien et al 2006, reviewed studies on sucrose consumption found that food intolerance to sucrose is less frequent than many other foods.

Low blood glucose levels, but higher than those that can be described clinically as hypoglycemic, is associated with irritability and violence. However, sucrose is not the predominant cause of swings in blood glucose levels.

Micro-nutrient decreased anti-social behaviour. Micro-nutrient intake is more closely associated with the total energy rather than sucrose intake; typically the amount of sucrose in the diet does not lead to micro-nutrient deficiency.

Benton concluded that studies that have examined the impact of sucrose on the behaviour of children produced no evidence that it has an adverse influence, such as attention deficit hyperactivity disorder.

However, sugar fat and salt-reduction should go on in face of the obesity and cardiovascular risks. Physical activity not also reduces risk of obesity, but also may improve mental health in adolescents.

Physical activity and mental health

Five to seven weekly hours of physical activity at age 15-16 years was weakly associated with mental health. The authors called for further studies about physical activity as a possible protective factor in relation to mental health problems in adolescence.

Sodium benzoate as source of benzene in soft drinks

Benzene is listed as a poisonous chemical shown to increase the risk of leukaemia and other cancers. Drinking water limits range from 10 parts per billion (World Health Organisation), 5ppb in the US and one part per billion in the EU. Some soft drinks contain up to five times this limit of benzene. The FDA assured benzene did not present an immediate health risk at the levels found to date in drinks.

Interaction among sodium benzoate and other ingredients

Mike Redman the National Soft Drink Association (NSDA) in a meeting of the FDA in 1990 suggests that benzene formation occurs as an interaction among ingredients in the product for example, sodium Benzoate, ascorbic acid and FD&C yellow Nr. 5 under certain conditions might produce benzene.

Other interaction between sodium benzoate and other additives

Other chemicals such as erythorbic acid, EDTA, oxygen and sweeteners like high fructose corn syrup influence benzene formation in the presence of benzoic acid.

Sweeteners are thought to inhibit the reaction, as the problem seems most noticeable in diet drinks. EDTA appears to inhibit the reaction also, possibly by complexing metal ions that could act as catalysts.
Companies added Calcium disodium ethylene diamine tetra-acetate (" EDTA") in the US to reduce benzene formation. The FDA, however noted that calcium disodium EDTA is not an approved food additive for non-carbonated soft drinks. [84]

Erythorbic acid may lead to benzene formation in much the same fashion as ascorbic acid. Sodium benzoate and ascorbic acid (vitamin C) are still used together in a wide range of soft drinks and flavoured waters across the world. They can react together to cause benzene formation.

Ascorbic acid initially reacts with metals, such as iron or copper, found in the water to create free radical. These hydroxyl radicals break down Sodium benzoate into benzoic acid and benzene.
Reactivity of Ascorbic acid in presence of ions of copper and iron has been studied by Hans Steinhart et col. in 1993. [85]

Photolytic degradation of sodium benzoate (E211) in flavoured mineral water

The State of Florida Laboratories found traces of benzene in Koala Springs flavoured mineral water. There is speculation that the benzene traces originated from the sodium benzoate by way of photolytic degradation. [83]

Benzene in foods without added benzoates

The combination of sodium or potassium benzoate with ascorbic acid was shown to produce low levels (ng/g) of benzene in fruit-flavoured soft drinks. The presence of benzene was also reported in butter, eggs, meat, and certain fruits; levels of these findings ranged from 0.5 ng/g in butter to 500-1900 ng/g in eggs.
Slightly higher levels were present in some foods and beverages containing both ascorbic acid and sodium benzoate. [86]

Soft drinks, juices, beers, and waters from processed vegetables were analysed for trihalomethanes (THMs), benzene, and toluene. The THMs, which include chloroform, bromodichloromethane, dibromochloromethane, and bromoform, are reaction by-products of water disinfection by chlorination.

In this study benzene residues were found typically <5 ng/g, except for 7 and 9 ng/g in 2 foods, compared to other contaminants: Toluene residues were typically <3 ng/g except for 23, 29, and 75 ng/g in 3 canned foods and chloroform was none detected to 94 ng/g in the 44 foods analysed. [87]

Industry tests on soft drinks found that temperatures of 30° and exposure to UV light for several hours were enough to more than triple benzene residues in some drinks.

America's soft drinks industry association said hot warehouses and cars parked in direct sunlight are examples of when soft drinks would be exposed to even higher temperatures as that.

Benzene formation may also occur in dietary supplements, such as liquid aloe vera and vitamin formulations, and other acidic liquid food products, such as lemon juice where sodium benzoate and ascorbic acid are present.

How to avoid benzene in food

Producers should use predictive testing to simulate storage under UV and heat up to 50° to survey their products for benzene.

The consumer should carefully read the list of ingredients. When sodium benzoate is listed in soft drinks and other acidified foods do not buy it.

Food regulations worldwide should demand sodium benzoate to be taken out of juices, soft drink and other acidified food formulas. Technology does not need sodium benzoate in drinks as modern heat sterilization and aseptic filling techniques make the use of this preservative unnecessary.

Belgian soft drinks with Benzene, benzoic acid and benzoates should be banned from acidic beverages

Christof Van Poucke and colleagues 2008 analysed 134 Belgian soft drinks. The authors found that ten samples were above the European limit for benzene in drinking water of 1 µg L, and one sample had a concentration of 10.98 µg /L, thereby exceeding the action limit for benzene in soft drinks of 10 µg/ L. The authors stress that benzene can be formed when benzoic acid, a food preservative, is combined with ascorbic acid acidity regulators and interaction with packaging materials in foods like soft drinks.

Benzene oxidises in the body to produce an epoxide, benzene oxide, which is not excreted readily and can interact with DNA to produce harmful mutations.

The use of preservatives like benzoic acid and benzoates can be avoided by sanitising filling lines and adopting sound hygienic strategies. Soft drinks and other beverages are very easy to handle in an sterile environment. Chemical preservatives are not needed in modern food technology.

Volatile organic compounds (VOCs)

VOCs are a group of low molecular weight aliphatic and aromatic compounds with low boiling points. Sources of VOCs include solvents, dry cleaning compounds, degreasers, paints, chemical intermediates, and assorted industrial products. They are also products of combustion and the chlorination of drinking water. Additionally, VOCs can come from the process of microwaving foods. Some VOCs are even allowed as indirect food additives from components of commercial packaging.

FDA study, published in 2003 a study of Fleming-Jones and Robert E. Smith. Volatile organic compounds (VOCs), such as benzene, were found in at least one sample of all foods tested, although no single compound was found in each of the foods. Benzene levels ranged from 1 to 190 ppb,

Benzene is a human carcinogen and neurotoxin. Other VOCs reported to be human neurotoxins include 1,1,1- trichloroethane, styrene, toluene, trichloroethylene, and xylene. However, the doses needed to cause neurotoxicity are far greater than those detected in foods in the study of Fleming-Jones. [90] [91] [92]

The amounts needed to cause these toxicities are much higher than those found in foods in this study. For all toxicities except carcinogenesis, a threshold approach is often used, which sets acceptable daily intake levels. However, it is sometimes stated that a nonthreshold approach is used for potential carcinogens and that a single molecular adduct can initiate the multistep process of carcinogenesis. [93]

FDA concluded from data derived from the FDA's Total Diet Study that the American food supply is comparatively safe. Although there is some oral exposure to VOCs, they are usually inhaled at much higher doses through cigarette smoke, gasoline fumes, and industrial pollution.

Sodium benzoate enhances detrimental effects of free radicals

Peter Piper proposes a Saccharomyces cerevisiae petri dish test, using yeast superoxide dismutase mutants to distinguish a compound that enhances the detrimental effects of endogenous reactive oxygen species production by the mitochondrial respiratory chain from another chemical that generates oxidative stress by redox cycling.

Using this test system, Piper found that weak organic acid food preservatives exert strong pro-oxidant action on aerobic yeast cells, and are mutagenic toward the yeast mitochondrial genome.
The author concluded that sodium benzoate may generate oxidative stress within the epithelia of the gastrointestinal tract.

ICBA Benzene Guidance

The International Council of Beverages Associations (ICBA) is a non-governmental organization representing the interests of the worldwide beverage industry.

Today, as the beverage industry continues to grow and expand, the International Council of Beverages Associations (ICBA) is renewing its commitment to provide guidance on preventing/minimizing benzene formation. This guidance will be made available to all beverage companies worldwide.

German assessment of benzene in foods

The Federal Institute for Risk Assessment has issued an expert opinion on whether benzene may be formed in beverages from the food additive benzoic acid in the presence of ascorbic acid.

It is advisable to minimise and/or avoid the intake of benzene as far as possible. This is the standard practice for substances which are to be considered as carcinogens and germ cell mutagens.

However, further chemical-analytical data are needed in order to assess the possible risk from simultaneous use of benzoic acid and ascorbic acid in foods. If it turned out that the simultaneous use of benzoic acid and ascorbic acid in foods makes a significant contribution to total benzene intake, it may be necessary to check whether the authorisation framework for a simultaneous use of benzoic acid and ascorbic acid in foods would have to be changed.

Brilliant Blue FCF E133

Other names: Other names(FD&C Blue No.1, Acid Blue 9, D&C Blue No. 4,Alzen Food Blue No. 1, Atracid Blue FG, Erioglaucine, Eriosky blue, Patent Blue AR, Xylene Blue VSG, C.I. 42090). [97]

This colouring agent has previously been banned in Austria, Belgium, Denmark, France, Germany, Greece, Italy, Spain, Sweden, and Switzerland among others but has been certified as a safe food additive in the EU and is today legal in most of the countries. E133 was allowed to be used in some foods in the European Union by the Regulation 1129/2011. [98]

It is allowed to be used in sweets, confectionery, dessert, ices, in some soft drinks, baked goods and tinned processed peas.
It has the capacity for inducing an allergic reaction in individuals with pre-existing moderate asthma. In the United States production exceeds 1 million pounds annually, and daily consumption is around 16 mg per person. An ADI of 0,1 mg/Kg bodyweight was determinated. In animals high amounts of brilliant blue E133 produced depositions of the colour at kidneys and lymphatic system.

Blue discoloration and death from FD&C Blue No. 1 used in enteral feeding solutions [99]

The US Food and Drug Administration (FDA) has advised that several reports of toxicity, including death, have been associated with the use of FD&C Blue No. 1 (Blue 1) in enteral feeding solutions. Reported episodes were manifested by blue discolouration of the skin, urine, faeces, or serum and some were associated with serious complications such as refractory hypotension, metabolic acidosis and death. Case reports indicate that seriously ill patients, particularly those with a likely increase in gut permeability (e.g., patients with sepsis), may be at greater risk for these complications.

Quinoleine Yellow

Quinoline Yellow is absorbed from the gastro-intestinal tract to only a small extent in rats and dogs, and most of an orally administered dose is excreted unchanged. No adverse effects of treatment were seen in the two-generation long-term study in mice. In particular there was no observed effect on thyroid function or histopathology and no evidence of carcinogenicity.
Quinoline yellow is banned in foods in Australia, the US and Norway. In UK it is used in sweets, smoked haddock, confectionery and pickles.


Level causing no toxicological effects: Mouse: 1% of the diet, equal to 1.500 mg/kg/
Estimate of an acceptable daily intake for man: 0-10 mg/kg bw.

The committee noted that there are two quinoline yellows, one of which is about 30% methylated and the other non-methylated, and considered that data from both compounds could be used for toxicological evaluation of either of the quinoline yellows for food additive use.[100]

Monosodium glutamate

Monosodium glutamate is banned in baby food in the UK, but is used in a variety of crisps and snacks which are consumed by children. Parents should watch the ingredient list carefully in order to avoid to give products with these ingredients to children under 5 years.

Natural cure for meats


Nitrates and nitrites

They are used in cured meats such as ham and bacon, hot dogs and deli meats. Both nitrates and nitrites combine with other nitrogen-containing substances in the stomach to form N-nitroso compounds that are known to cause stomach cancer.

Salted, smoked or pickled foods and red meat

Commonly preserved food by salting, smoking or pickling often contain large amounts of nitrites and nitrates. Countries where consumption of salted meat and fish and pickled vegetables is high, such as Japan and Korea tend to have correspondingly high rates of stomach cancer. Eating a diet high in red meat, especially when the meat is barbecued or well-done, also has been linked to stomach cancer.

Traditional curing methods using nitrite or nitrate are being substituted by new products for the sake of a natural label. There are consumers which try to avoid nitrite cured meat because of health concerns.

Curing as food preservation

In food preparation, curing refers to various preservation and flavouring processes, especially of meat and fish by the addition of a combination of salt, sugar and either nitrate or nitrite. Many curing processes also involve smoking.

Nitrite curing retards rancidity, stabilises flavour, and establishes the characteristic pink colour of cured meat.

Salt inhibits the growth of microorganism which cause spoilage by drawing water out of microbial cells through osmosis. As the unwanted bacterial population decreases, other beneficial bacteria, primarily of the Lactobacillus genus, come to the fore and generate an acidic environment (around 4.5 pH).

The sugar included in the cure is used as food by the lactobacilli; generally dextrose is preferred over sucrose, or table sugar, because it seems to be more thoroughly consumed by the bacteria. This process is in fact a form of fermentation and, in addition to reducing further the ability of the spoilage bacteria to grow, accounts for the tangy flavour of some cured products. Concentrations of salt up to 20% are required to kill most species of bacteria.

Smoking adds chemicals to the surface of an item which affect the ability of bacteria to grow, inhibit oxidation (and thus rancidity), and improve flavour.

Nitrates and nitrites not only help kill bacteria, but also produce a characteristic flavour, and give meat an appealing pink or red color. Nitrate (NO3), in the form of either sodium nitrate or potassium nitrate is used as a source for nitrite (NO2). The nitrite further breaks down in the meat into nitric oxide (NO), which then binds to the iron atom in the center of mioglobin's heme group, preventing oxidation.

Commercially Curing salt containing small amounts of sodium nitrite or sodium nitrate is used in the preserving and curing of meats, and in sausage making. The nitrate component inhibits the growth of bacteria, specifically botulism, and helps preserve the color of cured meat.
The presence of nitrates and nitrites in food is controversial due to the development of nitrosamines when the food, primarily bacon, is cooked at high temperatures. The nitrate and nitrite compounds themselves are not harmful, however, and are among the antioxidants found in fresh vegetables. [103]

The usage of either compound is carefully regulated in the production of cured products; in the United States, their concentration in finished products is limited to 200 ppm, and is usually found to be below. Finally, they are irreplaceable in the prevention of botulinum poisoning from consumption of dry-cured sausages.

Nitrite replacement for cured meats

Chr. Hansen has developed a new natural cure for manufacturers as a nitrite replacement for cured meats. No synthetic nitrates or nitrites are added, labelling of nitrite or nitrate can be avoided. The product can be labelled as 'uncured' in accordance with USDA 9CFR317.17 labelling regulations [101]. Naturally occurring nitrates are converted by the culture to nitrite which then accounts for the curing of the meat.

According to papers of Chr. Hansen various strains are used. Lactobacillus sakei BJ-33 was approved by the Danish authorities for bioprotective use. The bacteria multiplies at chill temperatures as low as 2°/35° F and suppresses the spoilage flora of indigenous lactic acid bacteria and Brochothrix thermosphacta. The inhibitory effect possessed by the strain is due to competition since the ability to produce bacteriocins has not been detected. It is used in combination with a strain of Staphylococcus xylosus.

Other strains used by Chr Hansen are Leuconostoc carnosum 4010 (formerly named Lc-1043). Another strain used is B-LC-20 which is an adjunct culture that is added on top of the existing starter culture without changing the sausage recipe or the sausage processing procedure for the reduction of Listeria in fermented dried sausage.

Processed meat like bacon and sausages increase risk of heart disease and diabetes, says a meta-analysis

Micha, Wallace and Mozaffarian 2010 found that Consumption of processed meats, but not red meats, is associated with higher incidence of CHD and diabetes mellitus. It takes only 50 g (one hot Dog) of processed meat per day to increase heart disease by 42% and type 2 diabetes by 19%. No link between eating unprocessed red meat like beef or pork and risk of heart disease and diabetes was found by the authors. High levels of salt and nitrate preservatives in sausages, bacon and deli meats, rather than fats, might explain the higher risk of heart disease and diabetes seen with processed meats, but not with unprocessed red meats.

The authors call for studies looking at processed and unprocessed meats separately and focus on salt and nitrate preservatives.

Researcher from Iran says that nitrite may be replaced by annatto up to 60 per cent

A controversy concerns nitrites in cured meats. Some say it increases the risk of stomach cancer, or lung diseases [107], while other researchers found no proof of such affirmations. Cardiologists found that nitric oxide from bacterial breakdown of nitrite reduces blood pressure and avoids damages during an heart attack. [108] [109]

Ribeiro and colleagues 2006 presents a summary of studies concerning anti-mutagenic and anti-carcinogenic potential of annatto, mushrooms and propolis in Brazilian natural diets. Their data shown a clear role for these compounds in preventing mutation and specific preneoplastic lesions. The authors stress that these agents may become a promising alternative for cancer prevention strategies. [110]

Petersson, 2008 suggests that nitrites may protect the stomach from ulcers. The author says that mouth bacteria reduce nitrate to nitrite. Nitrous oxide NO produced in the gastric lumen after nitrate ingestion increased gastric mucosal blood flow and the thickness of the firmly adherent mucus layer in the stomach. The blood flow and mucus layer are essential defence mechanisms that protect the mucosa from luminal acid and noxious agents. [111]

Zarringhalami, Sahari and Hamidi-Esfehani 2008, researchers from Iran, presented a study claiming that annatto (Bixa orellana L.) powder may replace up to 60 per cent of nitrite as ingredient of sausage. This might reduce excessive consumption of nitrite. According to the authors colour, flavour, aroma, and microbial contamination did not differ significantly compared with samples with 100 percent nitrite. [106]

Annatto is not allowed in the European Union as ingredient of sausages and other meat products. Annex IV of the European directive 94/36/EC on colours for use in foodstuffs cites annatto in a positive list of colours permitted for certain uses only, meat products are allowed there. [11]

Code of Federal Regulations 9CFR317.17

[101] With respect to sections 1(n) (7), (9), and (12) of the Act and Sec. 317.2, any substance mixed with another substance to cure a product must be identified in the ingredients statement on the label of such product. For example, curing mixtures composed of such ingredients as water, salt, sugar, sodium phosphate, sodium nitrate, and sodium nitrite or other permitted substances which are added to any product, must be identified on the label of the product by listing each such ingredient in accordance with the provisions of Sec. 317.2.

(b) Any product, such as bacon and pepperoni, which is required to be labeled by a common or usual name or descriptive name in accordance with Sec. 317.2(c)(1) and to which nitrate or nitrite is permitted or required to be added may be prepared without nitrate or nitrite and labeled with such common or usual name or descriptive name when immediately preceded with the term "Uncured" as part of the product name in the same size and style of lettering as the product name, provided that the product is found by the Administrator to be similar in size, flavour, consistency, and general appearance to such product as commonly prepared with nitrate or nitrite, or both.

(c)(1) Products described in paragraph (b) of this section or Sec. 319.2 of this subchapter, which contain no nitrate or nitrite shall bear the statement "No Nitrate or Nitrite Added." This statement shall be adjacent to the product name in lettering of easily readable style and at least one-half the size of the product name.

(2) Products described in paragraph (b) of this section and Sec. 319.2 of this subchapter shall bear, adjacent to the product name in lettering of easily readable style and at least one-half the size of the product name, the statement "Not Preserved-Keep Refrigerated Below 40 deg.F. At All Times" unless they have been thermally processed to Fo 3 or more; they have been fermented or pickled to pH of 4.6 or less; or they have been dried to a water activity of 0.92 or less.

(3) Products described in paragraph (b) of this section and Sec. 319.2 of this subchapter shall not be subject to the labeling requirements of paragraphs (b) and (c) of this section if they contain an amount of salt sufficient to achieve a brine concentration of 10 percent or more.

Nitrites and nitrates in cured meat fruit and vegetable may protect against heart attack

Sodium nitrite E250 produces the pink colour of cured meat. It also retards rancidity and stabilises the flavour.

The third National Health and Nutritional Examination Survey (NHANES) on 7,352 subjects over the age of 45, have suggested that increased consumption of nitrites from cured meat could increase the risk of lung disease. [107]

Other studies found nitrite and nitrate to be related to the incidence of cancer due to the formation of nitrosamines.

But recent research has found no convincing evidence that nitrite and nitrate pose a cancer risk. In reality they seem to protect heart cells from death after an heart attack.
The study published in the Proceedings of the National Academy of Sciences found that nitrites reduce heart cell death in mice by 48 per cent after an heart attack.

Nathan S. Bryan and colleagues report that mice supplemented with 50 mg nitrite per litre of drinking water for seven days had a survival rate of 77 per cent compared to 58 per cent for the mice without supplementation.

The researchers say that eating nitrite and nitrate rich foods such as fruits and vegetables and some meats in moderation can drastically improve outcome following a heart attack, They suggest to sty the effects of supplementation of nitrite/nitrate in the diet can decrease the incidence and severity of heart attack and stroke in patients with known cardiovascular risk factors.

Bryan suggests the formation of nitric oxide (NO) from nitrites to be the mechanism of the heart protecting effect of nitrates. Nitric oxide causes the cells of the blood vessels to signal the surrounding muscles to relax. This results in a reduction in blood pressure, reduction of blood clotting and and protection against myocardial infarction and strokes. A betaine in chemistry is any neutral chemical compound with a positively charged cationic functional group such as ammonium ion or phosphonium ion and with a negatively charged functional group such as a carboxyl group. Historically the term was reserved for trimethylglycine only. [112]

Betaine is found in high concentrations in sugar beet, improving consistency of pig and poultry performance improving the structure of the animals intestine, increasing gut tensile strength and the absorptive area for nutrients, which helps to maintain growth, feed conversion and profitability in the absence of antibiotic growth promoters.

Betaine (trimethylglycine) functions very closely with choline, folic acid, vitamin B12 and methionine. Betaine is necessary in the biochemistry of carnitine and serves to protect the kidneys from damage. [113]

Barak proposed in 1983 that betaine may serve as an important methylating agent when normal methylating pathways are impaired by ethanol ingestion, drugs or nutritional imbalances. Furthermore, betaine may prove to have therapeutic application in cases of altered folate, vitamin B12 or methionine metabolism. [114]

Glycine betaine (betaine hydrochloride) used as feeding attractant was found to have a positive effect on the growth and feed conversion of juveniles of Macrobrachium rosenbergii (deMan). Weight gain, feed intake and food conversion ratio (FCR) were higher in prawn fed glycine betaine-added diets compared with the control feed in a study made by Dr Nathan Felix. [115]

Betaine is used in animal feeds as a source of essential methyl groups and as an osmoregulant. Its osmotic function is useful in maintaining gut wall integrity. Feeding betaine could compensate the removal of antibiotic growth promoters. Reduced control of Clostridium perfringens in birds could be countert with betaine which improves the hydration and integrity of the gut wall. [116]

Betaine decays in the liver forming methionine. Dietary methionine is spared. saving feeding costs Feeding betaine saves methionine and choline costs Betaine also functions as osmoregulator. Betaine in feed or water can help reduce dehydration by facilitating water retention in the body in case of heat stress. [117]

Advances in genetics and feed ingredients such as betaine from sugar beets and enzymes have reduce feed costs and generate a higher percentage of lean meat.

High content of nitrate in beetroot reduces blood pressure

Amrita Ahluwalia and colleagues 2008 studied the benefits of high nitrate content of vegetables like beetroot. The authors suggest that the nitrate content of the vegetables is responsible for a reduction of blood pressure , and not by the antioxidant vitamin content as assumed in foregoing studies.

The authors say that drinking 500 ml beetroot juice a day reduces blood pressure significantly three hours after the consumprtion. The authors explain that the high nitrate content of the juice increases nitrate content of the saliva, where it is converted to nitrite by bacteria living on the tong. When swallowed the nitrite is transformed in nitric oxide NO in the stomach and can the reenter the blood circulation as nitrite.

The authors reported further that dietary nitrate load also prevented endothelial dysfunction induced by an acute ischemic insult in the human forearm and significantly attenuated ex vivo platelet aggregation in response to collagen and ADP.

Some studies suggest that nitrite (NO(2)(-)), is a physiological signaling molecule with potential roles in intravascular endocrine nitric oxide (NO) transport, hypoxic vasodilation, signaling, and cytoprotection after ischemia-reperfusion. There is evidence that nitrite mediates many of the systemic therapeutic effects of NO gas inhalation, including peripheral vasodilation and prevention of ischemia-reperfusion-mediated tissue infarction. [119] [120]

Michael P. Frenneaux and colleagues 2008 found nitrite to be a potent venodilator in normoxia and hypoxia. The authors highlight the importance of nitrite as a selective arterial vasodilator in ischemic territories and as a potent venodilator in heart failure. [121]

Other studies say that nitrate and nitrite are linked to stomach cancer and give advice to avoid food with high content of both substances. The Mayo Clinic names some factors which are believed to increase stomach cancer: [122]

Thickening agents from Algae

Carrageenans extracted from red algae and alginates from brown algae are already being used as stabilisers and thickening agents.

Albert Mihranyan and colleagues from Uppsala University in Sweden studied the properties of alpha - cellulose of the green algae Cladophora sp. They found that Cladophora cellulose powder could be a useful alternative to commercially available dispersible cellulose grades when very low quantities of stabilizing agents are required.

Cherry fruits or cherry extract may prevent recurrent gout attacks

[124] Researchers report that consuming up to three servings.cherries or cherry extract over two days, lowers the risk of gout attack.

Cherry intake during a 2-day period was associated with a risk reduction of 35% gout attaks, and cherry extract intake was associated with a risk reduction of 45%. The risk was even further reduced to 75% when cherry was used together with allopurinol. Zhang et al. 2012, authors of the study explain that cherries may decrease serum uric acid levels by increasing glomerular filtration or reducing tubular reabsorption. This is Cherries have The high levels of anthocyanins of cherries have anti-inflammatory activities.

The authors stress that recommendations to reduce the risk of gout attack include moderation in alcohol consumption, weight reduction, decreased high-purine foods from the diet and eating cherries. However, cherries should not replace medication but should be consumed additionally to allopurinol.

Anthocyanin profile of sour cherry [125]

According to Damar and Eksi 2012 the antioxidant capacity, total polyphenolic content and monomeric anthocyanin contents of juices of different varieties of sour cherries were within the ranges 20.0-37.9mmol/L, 1510-2550 and 350.0-633.5mg/L, respectively. The main anthocyanin compound in sour cherry juice was cyanidin-3-glucosylrutinoside and cyanidin-3-rutinoside.Anthocyanin capacity correlates with total polyphenol content, however no correlation was found between the antioxidant capacity and monomeric anthocyanin content.

Soft drinks, fructose consumption, and the risk of gout in men

A study by Choi and Curhan suggests that dietary fructose intake is a possible risk factors for gout increasing serum urate and is associated with hyperuraemia. Obesity, alcohol, and diet with the aree also associated with the onset of gout.

Hyon K. Choi and Gary Curhan, authors of the 12 years follow-up study found a strong association between sugar sweetened soft drinks and gout. They wrote that two servings a day of a sugar sweetened soft drink, high in fructose, increased the risk of developing gout by 85% compared with consumption of less than one serving of sugar sweetened soft drinks a month. Fructose rich fruits and fruit juices may also increase the risk. Diet soft drinks were not associated with the risk of gout. [127]

Other foods high in fructose are fruit-yoghurt, candies, backery where corn syrup is added.

Two or more soft drinks per week double pancreatic cancer risk

Sugar-sweetened carbonated beverages and juices have a high glycemic load relative to other foods and beverages. Mark Pereira and colleagues 2010 write two or more soft drinks per week, but not juices, may increase the level of insulin in the body double the pancreatic cancer risk compared with persons which do not drink soft drinks. However limitations of this study are seen in the fact that soft drink consumption is often associated with overweight increase, smoking, red meat intake and reduced exercise and may not be sufficiently considered in this study.

Interestingly South Chinese population differ from European and Western population regarding the effect of nutrition health related issues. Increased BMI is a robust risk factor for type 2 diabetes. This, however is not valid for South Asians which have relatively low BMIs despite a high prevalence of type 2 diabetes. Odegaard and colleagues 2009 found that Singaporean Chinese with lean or normal BMI have an increased risk of type 2 diabetes. [129]

Improving nutritional quality of chocolate products

A randomized controlled trial performed by Pase et al. 2013 provided evidences of positive effects of cocoa polyphenols on mood in healthy persons. However, cognitive performance remained inaltered during administration of cocoa polyphenols. The authors suggest further studies on cocoa polyphenols to ameliorate the symptoms associated with clinical anxiety or depression.
Robson 2013 deplores the high content of sugar in chocolate. Current chocolate bars have a high energy density of more than 2 kcal/g. He makes a plea to improve the nutritional value of chocolate bars which should be based on the nutritional value of the low energy dense late Paleolithic human diet to help reduce mental ill health, obesity, and other postprandial diseases. [130]

Robson stresses that the high energy density and low nutrient density that characterize the modern diet must be overcome simultaneously. People with a low-energy-dense diet (<1.6 kcal/g) have the lowest total intakes of energy, even though they consume the greatest amount of food. A processed food that is not both low energy dense and high nutrient dense is of poor dietary quality. Sugar in chocolate formulations should be replaced by natural non-caloric sweeteners, nanocellulose and calorie-free monk fruit extract (Siraitia grosvenorii) could be used to lower the energy density of chocolate. [131]

The Paleolithic diet: Degenerative non-communicable diseases are rare or nonexistent in hunter-gatherers eating a late Paleolithic diet, that is, a low energy dense diet with a wild plant-to-animal energy intake ratio 1:1, with fish and shell fish providing a significant proportion of the animal component. Nearly all the genes and epigenetic regulatory mechanisms humans carry today were originally selected for behaviourally modern humans who appeared in Africa between 100,000 and 50,000 years ago. Thus, it has been argued that the typical diet, physical activity patterns, and body composition of late Paleolithic humans remain normative for contemporary humans – and models for disease prevention recommendations. [132]

Bel-Serrat et al. 2012 report increased metabolic risk in 2 to 9 years old children with high intakes of solid hydrogenated fat and white bread, and low consumption of fruits, vegetables and dairy products.

High consumption of chocolate, nut-based spreads, jam, honey, sweets, soft drinks and manufactured juices increased significantly CVD risk. Children with high consumption of breakfast cereals presented a reduced CVD risk compared to those with little breakfast cereals intake. The authors stresses the role that diet may play on cardiovascular health, specifically clustered cardiovascular disease risk. [133]

Added sugars to total energy value of 12,35 is too high, compared to the current recommended levels, says the study of Colucci et al 2012 of children in the city of São Paulo, Brazil. Highest sugar consume was due to soft drinks followed by sugars (sucrose and honey) and chocolate powder. [134]

Adults consumed 9,1% of added sugar of energy intake and elderly 8.4%. Soft drink was the most important source of sugar among adults. Elderly consumed table sugar most frequently. Soft drink and table sugar were the source of more than 50% of added sugar wrtote Bueno et al. 2012.


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