Benzoate and Synthetic Color Risk Assessment of Fast Food Sauces Served at Street Food Joints of Lucknow, India
Krishn Kant Mishra,
Subhash K. Khanna
Quality compliance of benzoate and food colors in the
two common fast food accessories, namely tomato and chilli sauces, served
at Street Food Joints (SFJs) has been evaluated and compared with the
counterpart branded products. Both sauces served at SFJs employed excessive
amounts of benzoic acid in comparison with branded products (p<0.05).
In spite of ban, presence of artificial colors such as Amaranth, Carmoisine,
Erythrosine, Ponceau 4R, Sunset Yellow and Tartrazine were noted in a
vast majority of SFJ sauce samples. All branded sauces, however, adhered
to the prescribed limit of benzoate and contained no artificial color.
Intake estimates for benzoate and one of the colors, Sunset Yellow, in
high consumers among children are likely to saturate 33 and 20% of the
ADI at 95th percentile levels, respectively, which appears to be on higher
side considering intake from a single commodity. Street Food Joints (SFJs)
cater to the demand of floating domestic/foreign tourist populations and
serve as a source of employment generation. This sector needs some assistance
to enable them to survive in the competitive markets but not at the cost
of quality. Effective risk communication and food safety awareness measures
designed for street vending outlets are required so that such lapses of
street vending outlets could be minimized.
With the changing lifestyles, fast foods have become increasingly popular
among all sections of the society, Street Food Joints (SFJs) cater to
the demand of inexpensive foods for many poor urban house holds and floating
populations and serve as a source of livelihood/self employment generation
(Codjia, 2000). WHO has advocated that governments should formulate supportive
policies for improvements in the small-scale enterprises and street food
vendors (WHO, 2006a). It is felt that this sector needs some measures
of protection to enable them to survive in the competitive markets but
none-the-less at the cost of quality of food products. This is the crust
of recently launched program on Healthy Food Markets initiative (WHO,
2006b). The Ministry of Health and Family Welfare in India has commissioned
a survey on street foods and the findings indicate that this sector is
striving to attract more consumers, it appears to pay less attention in
meeting quality and safety norms (Gupta, 2006). Both tomato and chilli
sauces are the most common fast food accessories. The existing food legislation
covered under the Prevention of Food Adulteration (PFA) Act of India and
rules therein permit benzoic acid as one of the class II preservatives,
with a maximum allowable limit of 750 mg kg-1 in case of tomato
and other sauces (PFA, 2005). It is feared that some informal domestic
units and Street Food Joints adapt to measures of approximation and may
ignorantly add excess quantities of preservatives to extend shelf life
of the product. Also, some informal units tend to substitute tomatoes
by cheaper vegetable raw material, though the rules specifically prohibit
the use of other vegetables in tomato sauce/ketchup (PFA, 2005). For such
products, the processors then resort to use of synthetic colors to disguise
cheap vegetable sauce and offer it as tomato sauce. It is precisely for
such reasons that the existing rules do not permit use of any extraneous
synthetic color in tomato sauce/ketchup (PFA, 2005). Among the food processing
units, the organized sector producing branded products, invariably adhere
to the prescribed permissible limits of food additives while, the same
may not be always true with the informal domestic scale units or SFJs.
It was hence a matter of curiosity to monitor the use pattern of benzoate
and synthetic colors in tomato and chilli sauces vended at SFJs and compare
these with the branded products and undertake the health risk assessment
in the consumers.
MATERIALS AND METHODS
Benzoic acid was procured from BDH, India. The synthetic colors Amaranth,
Carmoisine, Erythrosine, Ponceau 4R, Sunset Yellow FCF and Tartrazine
were purchased from Hickson and Dadajee, Ltd., Mumbai, India. HPLC grade
acetonitrile and methanol were obtained from Merck Limited, Mumbai, India.
Analytical grade Potassium dihydrogen ortho phosphate and ExelaR grade
acetic acid and liquor ammonia (specific gravity 0.91) were the products
of Qualigen India. Whatman Number 1 chromatography grade paper was purchased
from Whatman International Ltd., Maidstone, England. All other chemicals
used were of analytical grade, available commercially.
The samples of tomato and chilli sauce were collected from local markets
around the city of Lucknow, India during 2006. Among branded products,
50 tomato sauce bottles of Maggi, Kissan (Hindustan Lever), Heinz, Nestle,
Everest and Tops and 43 tomato sauce sachets of Cremica, Kissan and Tops
and 25 chilli sauce bottles (Pouchong, Continental, Top-up, Samrat and
Goyal) were procured from local retail markets. In the non-branded category,
100 tomato sauce and 25 chilli sauce samples vended at Street Food Joints
A 2.5 g portion of the sample was extracted thrice using 5 mL of methanol
each time and centrifuged. The methanolic extracts were pooled and volume
was made up to 15 mL for determination of benzoic acid and synthetic colors.
Determination of Benzoic Acid
High Pressure Liquid Chromatography (HPLC) based AOAC (2000) method
was adopted for the detection and quantitation of benzoic acid in tomato
and chilli sauce samples. Waters LC module (Waters Associates, Vienna,
Austria) equipped with a dual pump (model # 510), Rheodyne injector with
20 μL loop and tunable absorbance detector (model # 486) were used,
employing the reversed phase C18 μ-Bondapak column (3.9x300
mm; Waters) with a pre-column of the same type. One milliliter of the
methanolic extract was filtered through 0.45 μm Millipore filter
and 20 μL was injected. A standard solution of benzoic acid (10 μg
mL-1) in methanol was used for comparison. The elution was
carried out at ambient temperature (25<°C) with isocratic solution
of 0.01 M phosphate buffer, pH 2.3: acetonitrile (6:4, v/v) at a flow
rate of 1.0 mL min-1. The measurement was carried out at 230
nm and retention time and peak areas were recorded. The chromatograms
were recorded and processed by Waters Millennium7 software.
The Limit of Detection (LOD) of benzoic acid is 5 mg kg-1.
As part of validation, spectral analysis of eluted fractions coming out
of HPLC column was performed and matched with that of standard benzoic
acid. The recovery of benzoic acid from spiked sauce samples was found
to be > 85%.
Separation, Identification and Quantitation of Colors
For detecting the presence of colors in sauce samples, 5 mL of methanolic
extract was concentrated to 1 mL. The colors were selectively picked up
on wool, eluted and resolved on paper chromatograms using the method followed
by Tripathi et al. (2007). The quantification of various colors
was undertaken by measuring absorbance at respective λmax
along with standards, employing double beam spectrophotometer (Perkin
Elmer Lambda Bio 20). The spectral characteristics of detected colors
were compared and matched with standard reference colors as a validation
measure. The values of detected colors have been corrected for their respective
mean recoveries from individual color spiked sauce samples.
Survey on Consumption of Tomato Sauce
The consumption pattern of tomato sauce in Lucknow area was assessed
by a limited household dietary survey through a Food Frequency Recall
method (Burdock, 1997). The 24 h dietary recall questionnaire was designed
to provide information about the name, age and sex followed by the questions
on the consumption of tomato sauce including amount and frequency of consumption.
A total of 60 families having 238 subjects (age 5-70 years) with 189 adults
(84 males and 95 females) and 59 children were surveyed. Average body
weight of children (30 kg) and adults (60 kg) has been drawn from National
Center for Health Statistics, USA (WHO, 1983). The consumption data was
recorded on Food frequency Questionnaire while interviewing with the subjects
and then data was transferred to Microsoft Excel Sheet. The mean value
represented the average consumers and that of 95th percentile, as high
consumers of tomato sauce and was used to calculate benzoate intake.
Student`s t-test was employed to calculate the significance of the differences
between branded and street food joints samples (Fisher, 1950). A p<0.05
was considered to be significant.
Analysis and Validation of Benzoic Acid and Colors
Figure 1A shows the HPLC profile of benzoic acid. The
retention time of standard and extracted benzoic acid from tomato sauce
samples matched at 7.55 min. The results were validated by taking the
spectra of the HPLC eluted peak of standard and extracted benzoic acid
showing a λmax at 222 nm (Fig. 1B).
The resolution of colors employed in sauce samples was carried out by
paper chromatography and the spectra of eluted colors was recorded and
matched with standard reference colors (Fig. 2). Some
samples confirmed the presence of colors such as Amaranth, Carmoisine,
Erythrosine, Ponceau 4 R, Sunset yellow FCF and Tartrazine.
Surveillance on Benzoic Acid
In the 93 branded bottled and sachet tomato sauce samples, the maximum
level of benzoate detected was 745 mg kg-1, which is within
the Indian permissible limit of 750 mg kg-1. All six samples
of one branded product contained no preservative while the minimum content
in others was 151 mg kg-1. The mean and the 95th percentile
values of benzoate for branded tomato sauce samples were 392±20.48
and 725 mg kg-1, respectively In case of tomato sauce samples
from SFJs, 59% exceeded the prescribed limit of benzoate. The minimum
and maximum benzoate levels detected were 353 and 2119 mg kg-1,
respectively. The mean and 95th percentile values for SFJ vended tomato
sauces were 903±44.26 mg kg-1 and 1636 mg kg-1,
respectively (Table 1). In case of chilli sauce, none
of the branded bottled samples exceeded prescribed limit of benzoate (with
a mean content of 556±28.64 mg kg-1) while all the samples
of chilli sauce vended at SFJs contained higher than permissible levels
of preservative. The respective minimum and maximum contents of benzoate
used were found to be 772 and 2104 mg kg-1. The mean and 95th
percentile values worked out to 1052±58.36 mg kg-1 and
1657 mg kg-1, respectively (Table 1). The
values of benzoate in tomato sauce samples vended at SFJs showed significant
change (p<0.05) when compared to branded samples.
||HPLC resolution profile (A) and spectral characteristics
(B) of standard benzoic acid (a) and tomato sauce sample having benzoic
||Spectral characteristics of individual standard food
colors (a) and extracted colors from tomato sauce samples (b). The
horizontal and vertical axis represents wavelength (nm) and absorbance
Surveillance on Synthetic Colors
None of the branded tomato sauce samples showed the presence of
synthetic colors, the use of which is legally prohibited in tomato sauce.
However, a vast majority (80%) of SFJs vended tomato sauce samples were
found to contain artificial colors (Table 2). Among
colors, blends of Sunset yellow FCF with Amaranth (172-742 mg kg-1),
with Carmoisine (148-597 mg kg-1) and with Ponceau 4R (154-825
mg kg-1) were most commonly encountered (79%). Individual colors,
Erythrosine (37-46 mg kg-1), Sunset yellow FCF (99-1624 mg
kg-1), Ponceau 4R (104-124 mg kg-1) and Orange II
(81 mg kg-1) were also encountered in some samples (21%)
(Table 2). Both Amaranth and Orange II even otherwise are not included
in the list of permitted food colors in India. In case of chilli sauce,
none of the 25 branded bottled products showed presence of artificially
added colors but 80% of the SFJs vended samples were found to contain
synthetic color, Tartrazine. The maximum quantity noted was 500 mg kg-1,
with the mean and 95th percentile values of 166 and 391 mg kg-1,
||Concentration profile of benzoate in tomato and chilli
sauce served at Street Food Joints (SFJs): Comparison with branded
|a: The permissible limit of benzoic acid
in tomato and chilli sauce is 750 mg kg-1 under the PFA
Act of India, *: p<0.05, when compared to branded products
||Presence of synthetic colors in tomato and chilli sauce
served at Street Food Joints (SFJs): Comparison with branded products
|a: Individual colors: Erythrosine (37-46 mg kg-1),
Sunset yellow FCF (99-1624 mg kg-1), Ponceau 4R (104-124
mg kg-1), Orange II (81 mg kg-1) were detected
in 21% samples while blends of Sunset Yellow FCF with Amaranth (172-742
mg kg-1), with Carmoisine (148-597 mg kg-1)
and with Ponceau 4R (154-825 mg kg-1) were detected in
79% of total analyzed samples, b: Only Tartrazine was detected
in between 93-500 mg kg-1 levels
Tomato Sauce Consumption Pattern in Local Household Consumers
A limited survey on tomato sauce consumption among 238 subjects
ranged between 10 to 50 g per intake. Majority of consumers were in the
category of 10-25 g (91%) while only about 9% consumers used higher quantities.
The mean (designated as average consumers) and 95th percentile (designated
as high consumers) tomato sauce consumption values come to 19 and 30 g,
on per intake basis, respectively (Table 3).
Intake of Benzoate Through Tomato Sauce
A conservative intake estimate of benzoate and the percentage saturation
of the ADI in average and high consumers of sauce at mean and 95th percentile
levels of benzoate are shown in Table 4. The intake
of benzoate through branded tomato sauce samples in children and adults
for average and high consumers ranged between 0.13 to 0.42 mg kg-1
b.wt. day-1, saturating the ADI values between 2.6 to 8.4%
only at the mean levels of benzoate detected in surveyed samples. At the
95th percentile levels of benzoate detected in surveyed branded tomato
sauce samples, the intake ranged between 0.25 to 0.81 mg kg-1
b.wt. day-1, with concomitant ADI saturation levels of 5.0
and 16.2% (Table 4). Since much higher levels of benzoic
acid have been detected in SFJs vended tomato sauce samples, the intake
among average and high children and adult consumers at 95th percentile
benzoate levels work to 0.52 to 1.64 mg kg-1 b.wt. day-1
which tends to saturate 10 to 33% of ADI through this commodity.
In case of children population, SFJs vended tomato sauce consumption even
at mean levels of benzoate in average and high consumers saturated the
ADI between 11.4 to 18.0% (Table 4).
Intake of Colors Through Tomato Sauce
The intake of colors from tomato sauce served at SFJs is given in Table
5. Among detected colors, Amaranth, Carmoisine, Erythrosine and Ponceau
4R have been found to saturate only 1.2-3.5% of their respective ADI limits
even in high consumers of tomato sauce in children population. However,
Sunset Yellow at 95th percentile detected levels appears to saturate 20%
of its ADI from this source(Table 5).
|| Limited household surveillance on tomato sauce consumption
pattern in Lucknow area
|a: The mean and 95th percentile consumption
values for 238 respondents work out to 19 and 30 g, respectively
||Intake of benzoic acid in average and high consumers
of tomato sauce and saturation of ADI
|a: ADI of benzoic acid has been taken as
5 mg kg-1 b.wt. day-1 (JECFA, 2001) and the
values in parentheses represent its % saturation, b: The
average body weight of children (30 kg) and adult (60 kg) is based
on National Center for Health Statistics, USA (WHO, 1983)
| Percent saturation of ADI limits by individual colors
in high consumers of tomato sauce among children population
Of more than 5000 fruit and vegetable processing units in India, only
13% are large-scale organized units and almost 75% are informal home or
cottage scale (Pai, 2001). The demand for fast food accessory sauces is
showing steady increase with growing popularity of fast foods among all
sections of consumers. Both organized (branded products) and informal
units catering to SFJs (as well as some SFJs themselves) are engaged in
tomato and chilli sauce processing. Though the former are known to strictly
adhere to quality specifications, the later face market compulsions to
sell foods at affordable, competitive prices for economic imperatives
which often result in their resorting to cheaper alternatives leading
to compromised quality of the products. Street food trade is considered
an important source of employment, particularly for poor strata of society
and serves as a poverty alleviation measure (Abdussalam and Kaferstein,
1993; Codjia, 2000). However, the perception that some street-food vendors
may lack appreciation for safe food handling continues to be looked as
an area of public health concern (WHO, 1996).
In the present study, quality compliance of the two most common fast
food accessories, namely, tomato and chilli sauces, served at SFJs revealed
that these employed excessive amounts of benzoic acid and a variety of
artificially added colors in majority of samples, whereas none of the
branded products exceeded the prescribed benzoic acid limit or showed
presence of extraneously added color. Variable amounts of benzoic acid
at 750 or 1000 mg kg-1 are allowed in tomato sauce/ketchup
by different agencies (EC, 1995; FDA, 1999; Codex, 2004; PFA, 2005). A
survey on Italian foods and beverages revealed that the content of benzoic
acid in most cases was lower than that prescribed technically (Luca et
al., 1995).Similar monitoring in UK revealed that only 4 samples out
of 122 exceeded the permitted statutory limit of the benzoic acid (MAFF,
The intake of benzoate for average consumers in France (Verger et
al., 1998), Norway (Bergsten, 2000), Brazil (Tfouni and Toledo, 2002)
and Korea (Yoon et al., 2003) were well within the ADI limits.
The estimates of total dietary intake of benzoate in China, Finland, France,
Spain, UK and USA, using different methodologies revealed saturation of
ADI in range of 9-28%. A conservative estimate of benzoate in the present
study, through branded tomato sauce, in children consuming 30 g sauce
(95th percentile) daily may saturate 8 and 16% of ADI at respective mean
and 95th percentile levels of benzoate detected. However, same subjects
will saturate 18 and 33% of ADI from the similar consumption of tomato
sauce served at SFJs, which appears to be on higher side considering intake
from a single commodity.
Addition of synthetic colors is not permitted in tomato sauce under
the existing rules in India (PFA, 2005), as technologically there is no
requirement. Artificially imitated tomato sauce made out of pumpkin (Cucurbita
pepo L.) pulp not having even a speck of tomatoes obviously works
out to be a cheaper substitute. A vast majority of SFJ vended tomato sauce
samples were found to employ single or blend of colors and contained Amaranth,
Carmoisine, Erythrosine, Ponceau 4R and Sunset Yellow FCF. A study by
Food Standard Agency, UK on color usage in sauces used for marinating
found two samples showing higher usage of Tartrazine and Carmoisine and
took immediate action to ensure that consumers are not exposed to high
levels of colors (FSA, 2000).
A vast majority of the ready-to-eat (RTE) foods prepared in the non-industrial
sector at Hyderabad, India, were found to employ colors exceeding statutory
limit and Sunset Yellow was most frequently encountered color after Tartrazine
(Jonnalagadda et al., 2004). Another study on exposure assessment
to various food colors in children conducted at same location, revealed
that intake of both Tartrazine and Sunset yellow exceeded the permissible
ADI limits (Rao et al., 2004). Similarly, the consumption pattern
of colored foods among children in Kuwait showed that Tartrazine, Sunset
Yellow and Carmoisine exceeded their ADI limits (Husain et al.,
2006; Sawaya et al., 2007). In the present study, Sunset Yellow
was the most excessively used color in tomato sauce and in high consumers
among children population it saturated 20% of the ADI from this single
commodity, which appears to be on much higher side. This is apparent from
a New Zealand Food Safety Authority study on highly colored foods preferably
consumed by children which indicated that the average and high consumer
dietary exposure estimates for all food colors were less than 5 and 15%
of the relevant ADIs, respectively (NZFSA, 2002).
Under street vended conditions, the common conceivable hazards include
microbiological, chemical and physical contamination. But serving artificially
colored vegetable sauce and use of higher quantities of preservative,
as found in the present study, either ignorantly or presumably to shield
handling lapses under street vended conditions, appears a part of intentional
market driven phenomenon where economic gains prevail. Thus, effective
risk communication (Tobin, 2006) and sector based quality awareness programs
should be intensified to enable the principles of food safety to become
The authors are grateful to Director, ITRC for his keen interest in the
present study. One of us (SKK) is thankful to Indian Council of Medical
Research, New Delhi, for the award of Emeritus Medical Scientist scheme.
The manuscript is ITRC Communication No. 2522.
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