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VIETNAM NATIONAL UNIVERSITY – HO CHI MINH CITY

HO CHI MINH UNIVERSITY OF TECHNOLOGY
Chemical Engineering Faculty
Department of Process and Equipment

FOOD SAFETY RISKS ASSOCIATED WITH

THE PRODUCTION OF ARTIFICIAL FOOD
COLOR ADDITIVES CONTAINING FOOD
PRODUCTS.
LECTURER : Assoc. Prof.Nguyen Thi Lan Phi.
STUDENT : Nguyen Thi Minh Thu – 1752523
Tran Hien - 1752195
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Table of content.
1. Chemical hazards of synthetic food color…………………………………3
1.1. Yellow No. 5 (tartrazine)…………………………………………..3
1.2. Cochineal extract and carmine……………………………………..3
1.3. Unapproved food additives and color additives…………….……...4
1.4. Chemical hazards due to misformulation…………………….…….4
1.5. Chemical hazards due to incorrect labeling of substances associated with food
intolerance or food disorder………………………………………..5
2. Physical hazards of synthetic food color ………………………………….6
3. Assessment of the safety of food colours …………………………………9
3.1 Illegal dyes for colouring and associated health risks…………….…11
3.2Analytical methods developed for the detection of synthetic food colours..12
4. Food colours – current issues……………………………………………..23
4.1 Synthetic colours…………………………………………………..…23
4.2 Dyes - illegal veterinary drugs……………………………………....24
4.3 Colour migration………………………………………………….….25
5. Conclusions and summary………………………………………………..26
6. References………………………………………………………………...28

List of table.
1. Quick Reference Guide for Common Sources of Physical Hazards……8
2. Number of import alerts for each color additive on “Detension without physical
examination and guidance of food containing illegal and/or undeclared color”
……………………………………………………………………………11
3. Detection method for synthetic food color ………………………………13
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1.CHEMICAL HAZARDS OF SYNTHETIC

FOOD COLOR
Some color additives are specifically prohibited from use in food because we have
determined that the chemical additive poses a potential risk to public health.
Examples of such color additives are coumarin, safrole, and FD&C Red No. 4
(Red No. 4) (FDA, 2015b). We consider a prohibited color additive to be an
unapproved food additive or color additive for the purposes of the PCHF
requirements and, thus, to be a chemical hazard. You should consult 21 CFR if you
have questions about the regulatory status or safety of a particular additive when
formulating your food products.
1.1.Yellow No. 5 (tartrazine) is a color additive subject to color certification
under section 721(c) of the FD&C Act. (21 U.S.C. 379e). People sensitive
to Yellow No. 5 can experience symptoms that range from mild to
moderately severe. For example hives occur in some intolerant individuals,
but in asthmatic individuals Yellow No.5 can trigger allergic type reactions
(including bronchial asthma). To help protect people who are sensitive to
Yellow No. 5, FDA’s regulation for Yellow No. 5 states that any food for
human use that contains Yellow No. 5 must specifically declare the
presence of the color additive by listing it as an ingredient (21 CFR
74.705(d)(2)). If Yellow No. 5 is added but is not declared, the product
would be both misbranded under section 403(m) of the FD&C Act (21
U.S.C. 343(m) and adulterated under section 402(c) of the FD&C Act (21
U.S.C 342(c)).
1.2.Cochineal extract and carmine: Cochineal extract and carmine are color
additives permitted for use in foods in the United States under conditions of
safe use listed in 21 CFR 73.100. For sensitive consumers, cochineal extract
and carmine can cause severe allergic reactions, including anaphylaxis (74
FR 207, January 5, 2009). Although the color additives cochineal extract
and carmine cause allergic reactions, they are not included in the eight.
Major food allergens identified in FALCPA. As a result, the color additives
cochineal extract and carmine are not included in the definition of “food
allergen” in part 117 and are not subject to the food allergen controls
specified in the PCHF requirements. In addition, FDA’s specific labeling
requirement in the color additive listing for cochineal extract and carmine
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(21 CFR 73.100(d)(2)), rather than the more general labeling requirements
of FALCPA, govern the food labeling requirements cochineal extract and
carmine. All human foods containing cochineal extract or carmine are
required to declare the presence of the color additive by listing its
respective common or usual name, “cochineal extract” or “carmine,” in the
statement of ingredients ((21 CFR 73.100(d)(2)). Additional information on
the labeling requirements for these two color additives can be found in FDA
industry guidance, Cochineal Extract and Carmine: Declaration by Name
on the Label of All Foods and Cosmetic Products That Contain These Color
Additives; Small Entity Compliance Guide (FDA, 2009a). Control
strategies for cochineal extract and carmine are similar to those applied to
food allergen labeling controls.
1.3 Unapproved food additives and color additives : A substance (other
than a food contact substance subject to a notification under section 409(h))
that is a food additive or a color additive must be used in accordance with a
food additive regulation permitting that specific use or a color additive
listing.
Some food and color additives are specifically prohibited from use in food
because we have determined that the chemical additive poses a potential risk
to public health (see 21 CFR part 189 and 21 CFR 81.10). Examples of such
food and color additives are coumarin, safrole, and FD&C Red No. 4 (Red
No. 4) (FDA, 2015b). We consider a prohibited food additive or color
additive to be an unapproved food additive or color additive for the purposes
of the PCHF requirements and, thus, to be a chemical hazard. You should
consult 21 CFR if you have questions about the regulatory status or safety of
a particular additive when formulating your food products. An additional
resource for you is the Food Additive Status List on our website (FDA,
2014b).
1.4 Chemical hazards due to misformulation:A food ingredient can be a
chemical hazard if it is added in excess of a maximum use level, regardless
of whether the maximum use level is established due to food intolerance
(such as for sulfites) or is otherwise a condition of safe use of a food
additive, color additive, or GRAS substance. Control strategies to prevent
misformulation of substances generally include process controls to ensure
that excessive amounts are not added.
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1.5 Chemical hazards due to incorrect labeling of substances associated

with food intolerance or food disorder: Although the mechanisms
whereby persons experience food intolerance or food disorder are different
from the mechanisms that cause food allergy, reactions due to food
intolerance or food disorder can cause significant health problems for those
affected, and the principal means that consumers have to avoid the
symptoms of food intolerance are the same means that consumers use to
avoid symptoms of food allergy – i.e., avoid foods containing the substance
that causes the problem. For example, people who are intolerant to lactose,
a sugar that is a component of milk, avoid food products containing milk to
avoid the symptoms associated with lactose intolerance. In addition, people
who have celiac disease avoid food products containing wheat and other
sources of gluten. Undeclared substances associated with a food intolerance
or food disorder are chemical hazards that can get into food because either:
(1) The food manufacturer did not properly declare the substance on the
product label; (2) unintended (and, thus, undeclared) substances are present
in a food due to incorrect labeling. Control strategies to prevent incorrect
labeling of substances associated with a food intolerance or food disorder
are analogous to those used to prevent incorrect labeling of food allergens
and, thus, you may find Chapter 11—Food Allergen Controls helpful in
preventing incorrect labeling of substances associated with a food
intolerance or food disorder. The preventive controls in that comprehensive
guide to food allergen control do not explicitly address substances
associated with food intolerance or food disorder, but may nonetheless be
useful in addressing chemical hazards due to incorrect labeling of such
substances
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2.PHYSICAL HAZARDS OF SYNTHETIC

FOOD COLOR
You must conduct a hazard analysis to identify and evaluate known or reasonably
foreseeable physical hazards (such as stones, glass, and metal fragments).
Physical hazards are broadly classified as “hard/sharp” physical hazards and
“choking” hazards. Both categories can cause injury to the consumer. These
injuries may include dental damage, laceration of the mouth or throat, laceration or
perforation of the intestine, and choking and may even lead to the death. Because
physical hazards cover a broad range of contaminants, such as glass, metal, plastic,
wood, and stones, such contamination can occur throughout the processing facility,
including the receiving dock for ingredients and supplies.
• Metal: Metal-to-metal contact during processing can introduce metal fragments
into products. For example, metal fragments can break off during mechanical
cutting and blending operations, and some metal equipment has parts that can
break or fall off, such as wire-mesh belts. FDA’s Health Hazard Evaluation Board
(FDA, 2005e; Olsen, 1998) has supported regulatory action against products with
metal fragments of 0.3 inches (7 mm) to 1.0 inches (25 mm) in length. Such
fragments have been shown to be a hazard to consumers. Metal hazards can be
controlled by the use of metal detection devices or by regular inspection of at-risk
equipment for signs of damage.
• Glass: Glass fragments can be introduced into food whenever processing involves
the use of glass containers. Normal handling and packaging methods, especially
mechanized methods, can result in breakage. Ingesting glass fragments can cause
injury to the consumer. FDA’s Health Hazard Evaluation Board has supported
regulatory action against products with glass fragments of the same size noted for
metal. Most products packed in glass containers are intended to be a ready-to-eat
(RTE) commodity. In your hazard analysis, you should consider the potential for
glass fragments to originate from sources other than glass containers used in
packaging. For example, some facilities that do not pack in glass prohibit the
presence of glass in the production environment to reduce the risk of glass getting
into the product. You can address glass fragments originating from sources such as
overhead light fixtures through CGMPs.
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• Hard Plastic: Hard plastic can be introduced into food when tools and equipment
such as scoops, paddles, buckets or other containers develop fatigue, crack, and
break as they wear. Hard plastic also can be introduced into food when plastic
sieves and screens deteriorate. You should examine items to determine whether
they are worn and remove worn items before they break, especially if they cannot
be effectively cleaned (e.g., because of small cracks).
In general, there is overlap between facility-related physical hazards and process-
related physical hazards. For example, equipment that has food-contact surfaces
that break during food processing and result in physical debris being deposited in
the food product can be considered a facility-related physical hazard (because the
equipment is part of the facility) or a process-related physical hazard (because the
equipment broke during processing). In general, in evaluating the potential for
physical hazards in your food products, it does not matter whether you consider
physical hazards to be facility-related or process-related. However, a few physical
hazards can readily be classified as facility-related or process-related. For example,
nuts and bolts used during maintenance procedures would be a facility-related
hazard, but production equipment that has nuts and bolts that could fall out during
production would be a processrelated hazard.
Table 3-9 is a Quick Reference Guide to help you identify common sources of
these physical hazards. See Chapter 13 – Preventive Controls for Physical Hazards
for more detailed recommendations on control measures for physical hazards. In
this guidance, we do not discuss ingredient-related physical hazards such as wood
and stone, which are usually addressed through CGMPs or as a supply-chain
control through your supplier program.
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Table 1. Quick Reference Guide for Common Sources of Physical Hazards

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3.ASSESSMENT OF THE SAFETY OF FOOD

COLOURS
The specific conditions for colours state that it should also serve one of the
following functions: restore the original appearance of the food, make food more
appealing and add colour to colourless foods. The additive must be subject to
toxicological testing, evaluation and re-evaluation in the light of new scientific
data. The Regulation specifies the foodstuffs to which the additive can be added
and the level of addition, taking into accounts the Acceptable Daily Intakes
(ADIs). ADI is ―an estimate of an amount of a substance in food or drinking
water that can be ingested daily over the lifetime without appreciable risk,
expressed in mg per kg of body weight.
The safety evaluation of food additives for human consumption requires a review
of published scientific studies on the toxicity of the additive such as animal studies
using large doses of additives for long periods to show that the substance will not
cause harmful effects at the expected levels of human consumption. The additive is
permitted if according to the current state of knowledge there is a reasonable
certainty of no harm to the consumer from ingestion of this additive under its
proposed conditions of use. The assessments of safety of food additives are based
on available scientific data on the toxicity of the chemicals that are collected and
evaluated by International Agency for Research on Cancer (IARC), National
Toxicology Program of the US Department of Health and Human Services (NTP)
or Joint Food and Agriculture Organization of the United Nation/World Health
Organization Expert Committee on Food Additives (JECFA).
The effect of the re-assessment can result in the change of the legal status of the
food additive. An example of such a re-assessment can be demonstrated by the
case of red 2G.Following the EFSA re-evaluation of the dye red 2G in 2007
(EFSA, 2007), Commission Regulation 884/2007 suspended its use. This dye was
permitted previously for colouring breakfast sausages; however there was
sufficient evidence that it is metabolized into the carcinogenic aniline to suspend
its use. Another dye brown FK (previously permitted in kippers only) has been
recently removed from the list of approved colours included in the Regulation(EC)
1333/2008 due to the fact it was no longer used and there was no sufficient data on
is safety available.
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3.1 Illegal dyes for colouring and associated health risks

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Dyes used for industrial applications, under some circumstances, can be

fraudulently added to a range of foods to increase their value i.e. when the colour
of the product is one of the indicators of its quality or to mask low quality of a
product. These dyes are cheap to produce and readily available. A number of
illegal dyes have been found or have been suspected of being present in foods
consumed in the EU (RASFF online database), United States(F&DA, 2013b), India
(Tripathi et al., 2007) or Japan (Ministry of Health Labour and Welfare, Japan)
with some examples (illegal in the EU) presented in Fig 3. Most of these dyes are
considered to be genotoxic and/or carcinogenic or there is limited data available to
conclude if they are safe or not. The two main groups of dyes: azo-dyes and
triphenylmethanes have been identified as those most likely to be illegally added to
food (EFSA, 2005). Azo dyes are a group of organic compounds widely used in
industry. The toxic activity of these dyes is a result of their metabolism (Golka et
al., 2004). The azo reduction (breakage of azo bond) is enzyme-mediated and
results in formation of active aromatic amines that can form DNA adducts. The azo
reductive enzymes present in liver, skin and gastrointestinal microflora appear to
play the main role in thein vivo reduction (Xu et al., 2007). The metabolism of the
azo-dyes is reduced by substitutions in the aromatic ring – sulphonation or
carboxylation and substitution of the hydrogen of an amino group. Substitutions
can reduce genotoxicity by decreasing lipid solubility of the dye and as are sult its
absorption (EFSA, 2005). Therefore sudan I is assumed to be carcinogenic whilst
sunset yellow FCF is a legal food additive. The only difference between these two
chemicals is the presence of two sulpho groups in the sunset yellow FCF structure
what causes a decrease in lipid solubility. The potential of a chemical to be
metabolized into a lipid soluble aromatic amine has been considered to be an
indication of genotoxic and/or carcinogenic action. Thus several dyes presented in
Figure 3. have been identified to be potentially harmful (EFSA, 2005). According
to the available toxicity data some of the illegal azo dyes that have been detected
recently in food sin the EU are possible genotoxic and/or carcinogenic agents.

3.2Analytical methods developed for the detection of synthetic food

colours
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Multiple chromatographic methods with different detection systems,

spectrophotometric, voltammetric and capillary electrophoresis techniques have
been developed for the analysis of synthetic colour compounds in foodstuffs; some
examples are presented in Table 4.
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14
15
16
17
18
19
20
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Reviews on the detection of natural colours (Scotter, 2011b) and sudan I-IV
specifically (Rebane et al., 2010)have been presented elsewhere. The multianalyte
HPLC-DAD method developed by Yoshioka & Ichihashi (2008) can be used for
the analysis of 40 synthetic legal and illegal colours in drinks and confectionary.
Other HPLC-DAD methods were developed to monitor the presence of 17 legal
and illegal (Bonan etal., 2013), 14 legal (Kirschbaum et al., 2003), 13 legal
(Minioti et al., 2007) or 7 legal (Harp etal., 2013) synthetic food colours. Tandem
mass spectrometry based detection has also been applied in the dye analysis field,
mainly to determine the presence of illegal and potentially harmful dyes. The
HPLC-MS/MS method developed by Feng et al. (2011) can be used for the
detection of 40 synthetic dyes in soft drinks and Zhao et al., (2012) for the
monitoring of the presence of 23 illegal dyes; Sun et al. (2007) of 10; Botek et al.
(2007) of 8 and Fang et al.(2013a) of 7. HPLC-MS/MS was also developed for 13
triarylmethane and phenothiazine dyes(Tarbin et al., 2008). Recently Liu et al.
(2011) used UPLC-MS/MS for the determination of 15illegal dyes. A number of
antibody-based screening methods i.e. enzyme-linked immunosorbent
assays(ELISA) were developed for the analysis of limited number of illegal
synthetic colours: sudan dyes, para red, orange II, triphenylmethanes (malachite
green, crystal violet, brilliant green),rhodamine B, methyl yellow and chrysoidine.
ELISA tests were also established for the detection of legal synthetic dyes – sunset
yellow and tartrazine.
Generally no safe level of the presence of illegal dyes in foods has been
established; however the FSA in the UK has recommended a limit 0.5 ppm for
products containing illegal dyes to be withdrawn from the market (FSA, 2007).
With regards malachite green in fish, the minimum required performance limit
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(MRPL) for the method used for the detection of the sum of malachite green and
its leuco metabolite is set at 2 ppb in the EU according to Commission Decision
2004/25/EC.
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4. FOOD COLOURS – CURRENT ISSUES

4.1 Synthetic colours
As a result of large number of cases contamination of food with sudan dyes in the
EU in2003 the emergency measures were taken and all chilli, chilli products,
curcuma and palm oil imported into the EU had to be screened for the presence of
sudan I-IV dyes. In 2005 and 2006 the Food Standards Agency (FSA) in the UK
tested 893 samples of spices, sauces and oils for sudan dyes, para red, rhodamine
B, orange II, red 2G, methyl yellow and metanil yellow and found six to be
contaminated (FSA, 2006). Additionally, 18 samples contained
annato/bixin/norbixin (E160b) a natural food colour that should not be used in
spices. Our own study on the application of the ELISA method for the detection of
sudan I has shown presence of this contaminant in 3 samples (out of 85 tested) that
were purchased within the EU (1 sample) and outside (2 samples).
A number of commercial companies offer screening services for several synthetic
dyes. However, there is still a need for the development of multianalyte methods
that could target a large range of suspect compounds. Such trends that are currently
observed in mycotoxin analysis (Varga et al., 2013) and veterinary drug residues
(Smith et al., 2009). Further expansion of the existing tools for fast, low cost
screening methods such as ELISA (limited to sudan dyes, parared, methyl yellow,
rhodamine B, orange II, chrysoidine and triphenylmethane dyes thus far)would
serve to provide further data on the occurrence of these contaminants in foods. The
list of possible targets is close to impossible to determine as there is such a large
number of synthetic dyes used in industry and widely available nowadays. New
dyes are being detected in food, for example in 2012 Ruf et al. reported the
presence of common industrial azo-dye basic red 46 in sumac spice sample during
the routine analysis for sudan dyes. Studies performed in countries from which the
dye-contaminated food is often imported can provide some background on the
targets that the analytical methods should focus on. One such study in India
reported the presence of a number of illegal dyes in coloured food commodities
purchased from urban an drural areas (Tripathi et al., 2007). Thirty one percent of
samples were found to contain illegal dyes: metanil yellow, rhodamine B, orange
II, sudan dyes, auramine, malachite green, amaranth and quinoline yellow (the last
two are not permitted in India), with more contaminants found in rural area (38%
of the total samples, while 25% in urban area) probably due to the less frequent
sampling and analysis of samples. This high level occurrence of illegal synthetic
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colours shows the potential scale of the problem in Asian countries and indeed
across the Developing World.
Dyes can be also added illegally to food to compensate for their low quality. In a
recent study the synthetic dye E154 was found in 9 out of 51 fishery products that
should not contain this dye according to the EU legislation. Carcinogenic
chrysoidine has been demonstrated to be used illegally as dyes in soybean milk or
to dye low-cost fish to imitate expensive and superior in quality yellow-fin tuna.
Chrysoidine has also antimicrobial and antispectic activities and as such it has been
applied to disinfect fish skin in some Asian countries.
4.2 Dyes - illegal veterinary drugs
Monitoring programs for carcinogenic triphenylmethane dyes: malachite green and
crystal violet are already in place in Europe and the US. However, there are a
number of dyes belonging to this group with similar properties that can be used as
alternatives and may not be detected in these monitoring programmes. Tarbin et
al., 2008 has suggested eleven other compounds belonging to triarylmethanes,
xanthenes and phenothiazine groups that should be under suspicion i.e. brilliant
green, ethyl violet, pararosaniline, victoria blue B, victoria blue R,victoria pure
blue BO, rhodamine 6G, methylene blue, azure B, new methylene blue and
nileblue. One of these compounds – victoria pure blue BO appeared on RASFF in
2010 as it was found in fish imported from Vietnam.
4.3 Colour migration
The migration of a colour from the package material or other food contact
materials has been recorded on RASFF since 2005 (RASFF online database).
RASFF does not generally give details about the identity of the dye that was
detected in food as a result of the migration. These are most probably dyes used in
industrial applications e.g. dying cutlery, napkins or wrappings and should not be
consumed by humans. A recent study demonstrated the migration of synthetic
colour brilliant green from green paper towels (Oplatowska et al., 2011c). Brilliant
green is atriphenyl methane dye that is likely to have similarly toxic properties as
the known illegal veterinary drug malachite green. A measurable amount of the
paper towel dye was shown to migrate from the towel trough the skin when the
towel was used for hand drying. Additionally, it was also demonstrated that large
amounts of colour can accumulate in food (fish was used as the model) when
towels containing dye were used for wrapping. This study raised a concern about
the safety of use of dyed paper towels for hygiene purposes and in food preparation
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area. A previously unidentified mean of exposure to legal synthetic colours was

demonstrated in a study performed by Lucova et al., 2013. Patent blue V (E131)
and brilliant blue (E133) were shown to directly enter the epithelium and then
possibly the bloodstream through the tongue from the saliva when used for
example in lollipops. These dyes could also penetrate the shaven skin when used in
aftershave products. This type of exposure does not include gastrointestinal tract
degradation and result in direct access of the dyes to the systemic circulation. The
authors concluded that due to the systemic availability these compounds should not
be used in topical products intended for use on slightly damaged skin and for
confectionary and lollipops for licking. Currently E131, E133 and other synthetic
dyes are commonly used in cosmetics, oral mouthwashes, pharmaceutical lozenges
and confectionary.
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5. CONCLUSIONS AND SUMMARY

The use of synthetic colours in food has resulted in a substantial amount of
negative press in recent years due to the possible harmful side effects. One of the
most important issues concerning the legal synthetic food colours is that there is
still no definitive answer if these dyes play any role in hyperactive behaviour in
children. However, due to the current suspicions there is a trend to remove them
from the food supply chain. The voluntary withdrawal action in the UK supported
by the Food and Standards Agency has already resulted in major food suppliers
changing from synthetic colours to natural products, especially in confectionery.
However, there placement is not always straightforward and not all natural colours
are completely safe.
Food globalization has resulted in humans consuming foods from many different
parts of the world. Another important issue is the lack of uniform regulations on
food colours what causes disruption to the international trade. Additionally, due to
the lack of uniform control policies worldwide, the distinct possibility of illegal
and harmful chemicals being present in foods locally produced or indeed imported
cannot be ruled out and indeed is quite likely, particularly in the Developing
World. The sudan scare in 2003 and contamination of hundreds of products with
illegal sudan dyes demonstrates the need for rigorous monitoring programs and
identifying and targeting of possible hazards. Currently, a number of analytical
methods have been developed for the analysis of food colours and a number of
companies across Europe and the US provide testing services for detecting illegal
dyes present in foods. Nevertheless, there is still a clear need for additional
research and monitoring activities to better protect the consumer. Therefore,
another challenge in the area of food colours is identification of known and
emerging dye adulterants/contaminants in food, not only in the Developed World
but as a matter of priority in the Developing World is needed. To support this need
the availability of rapid and low cost multicolourant screening and confirmatory
methods is identified as a priority.
Further exposure assessments of legal synthetic colours, especially in children are
required as a matter of priority to better understand the link between consumption
and adverse health effects. Without this data the ability to protect the consumer,
most especially the most vulnerable (children) is highly compromised.
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