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Analysis of the Concentration and Formation Behavior of Naturally Occurring

Formaldehyde Content in Food

Conference Paper · March 2017

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 Analysis of the Concentration and Formation
Behavior of Naturally Occurring Formaldehyde
Content in Food
Farrhin Nowshad
Department of Chemical Engineering, Bangladesh University of Engineering & Technology, Dhaka-1000, Bangladesh
Email: [email protected]

Md N. Islam, and Mohidus S. Khan

Department of Chemical Engineering, Bangladesh University of Engineering & Technology, Dhaka-1000, Bangladesh
Email: [email protected], [email protected]

Abstract ̶ In recent years, in spite of being legally prohibited, which is used as a biological preservative [3]. Recently, it
formaldehyde is reported to be widely used as a food has been reported that formalin is widely used in different
preservative to increase the shelf life of fruits and fishes in countries as a chemical preservative for fruits and fishes
tropical countries. The hot and humid weather of the [3]-[10]. There are direct and indirect health hazards
tropical countries tends to quickly deteriorate fruits,
associated with formaldehyde consumption. Consumption
vegetables, fishes, meat and other food items. Formaldehyde
is detrimental to human health and reported as a human of formalin on a regular basis can be injurious to the
carcinogenic. To prohibit formaldehyde application in foods, nervous system, kidney and liver, and may cause asthma,
the regulatory bodies often conduct on the spot analytical pulmonary damage and even cancer [5], [11]-[13]. The
tests to detect artificially added formaldehyde in food items. use of formaldehyde as a food preservative is prohibited
However, formaldehyde is ubiquitous in the environment in most of the countries [5], [14]. To restrict the use of
and is present in many animal and plant species as a formaldehyde as a food preservative, the regulatory
product of their normal metabolism. Formaldehyde can be bodies often collect food samples from local markets to
found naturally in food items including fruits, vegetables, perform on the spot analysis, or to send it to the nearby
meats, marine fishes and crustaceans. The formation and
analytical laboratory for the qualitative and quantitative
concentration of natural occurring formaldehyde may vary
according to food types and conditions. The naturally analysis of formaldehyde added in food items [6], [15].
occurring formaldehyde may interfere in the detection of However, formaldehyde is naturally present in a wide
artificially added formaldehyde in foods. It is therefore variety of food items, such as: fruits and vegetables,
important to study the concentration and formation meats, fish, crustacean and dried mushroom, as a
mechanism of naturally occurring formaldehyde in food common metabolic by-product [16]. In biological systems,
items. The objective of this study is to determine naturally formaldehyde is generated from different methylated
occurring formaldehyde levels in different fruits, vegetables compounds by demethylases, and from serine-glysine
and (cow) milk samples. In addition, time dynamic behavior interconversion catalyzed by pyridoxal phosphate [17].
of the formation of endogenous formaldehyde content in
Naturally occurring formaldehyde content also varies
food sample (banana sample; AAB genome of Musa Spp.)
was studied in this study. according to the food items and food conditions. The
presence of naturally occurring formaldehyde in food
items may interfere in detecting artificially added
Index Terms ̶ formaldehyde, methylated compounds, cancer
Risk, formation dynamics1 formaldehyde in foods. Therefore, it is important to
quantify the naturally occurring formaldehyde content in
food items to estimate external formaldehyde dosage.
I. INTRODUCTION This study aims to determine and quantify naturally
occurring formaldehyde content in food items such as
Formaldehyde is a flammable, highly reactive and fruits, vegetables and milk. The formaldehyde contents of
readily polymerizing colorless gas at normal temperature fresh fruits, vegetables and milk samples were
and pressure. It has a pungent, distinct odor and may determined using Nash’s reagent and spectrophotometer
cause a burning sensation to the eyes, nose, and lungs at [7], [8], [18]. In addition, the time dynamic behavior of
high concentrations [1], [2]. Formalin, a formaldehyde- naturally occurring formaldehyde formation in the food
water solution (40vol% or 37wt%), is a colorless liquid sample was analyzed. This study will help consumers and
regulatory agencies by providing baseline values of
naturally occurring formaldehyde contents in foods, and
Manuscript received November 8, 2016; revised Feb 16, 2017; also will help them to understand the dynamic behavior
accepted November 30, 2016.
Corresponding author: [email protected]
of formaldehyde formation in foods.
 II. POSSIBLE HEALTH HAZARDS OF FORMALDEHYDE experiments. Ultra-high purity de-ionized water
CONSUMPTION (18.2MΩ.cm, Purite, UK) was used for dilution.
Whatman 42 filter paper was used to filter the sample
The maximum allowable limit of formaldehyde in food
solutions. Nash’s Reagent was used as an indicator to
is 100 ppm [6]. If consumed at a higher concentration,
detect the absorbance (415nm) of formaldehyde in
formaldehyde may cause damage to the gastro intestinal
sample solutions [7]. Nash’s reagent was prepared by
(GI) tract, kidney, liver and lungs, and may lead to cancer
diluting 18.75g ammonium acetate in a 100ml erlenmeyer
[2]-[19]. Formaldehyde, when ingested, exerts an irritant
flask with an addition of 0.38ml of acetic acid and 0.25ml
action upon mucous membranes, and after prolonged use
of acetyle acetone. The total volume was made 62.5ml by
appears to cause inflammatory changes in the liver and
adding de-ionized water. Nash’s Reagent is light
also in the kidneys, where a portion of it is excreted [20].
sensitive, hence, it was kept in an air tight dark-glass
Furthermore, there are sufficient evidence linking
reagent bottle at room temperature [7, 10]. Fresh Nash’s
formaldehyde with nasopharyngeal cancer [5], [21]. The
reagent was prepared for every experiment. 0.1N
international Agency for Research on Cancer (IARC) has
potassium hydroxide and 0.1N nitric acid were used to
classified formaldehyde as well as formalin as a Group 1
adjust the pH of the distillate to be in range of 6.0 to 6.5
carcinogenic (Carcinogenic agents with sufficient
[7]. A calibrated Hanna Pocket Sized pH Meter Hi96107
evidence in humans) [3], [22]. Table I describes different
was used to check the pH and a Shimadzu UV-VIS 2600
health hazards caused by formaldehyde consumption.
spectrophotometer was used to measure the absorbance.
TABLE I. HAZARDOUS EFFECTS OF FORMALDEHYDE CONSUMPTION [2, 3, C. Sample Preparation and Formaldehyde detection
5, 13, 19]
Fruit and vegetable samples were peeled off, cut into
Ingestion Excessive ingestion can cause -
small pieces, and blended with water in 1:10 ratio. Using
 Severe pain with inflammation, ulceration and necrosis
of the mucous membranes lining almost every internal a clean cloth as sieve; the fruit juice was separated from
organ; the residual solids and then filtered using Whatman 42
 Nausea; filter paper. UTH milk samples were used directly and
 Vomiting blood;
 Diarrhea with bloody stool;
powdered milk samples were prepared by diluting solid
 Blood from urine; powder with water in 1:2 ratio; the milk sample was then
 Gastrointestinal lesions; filtered. The filtrate (fruits, vegetables and milk) was
 Acidosis; diluted 100 times. The pH of the diluted samples was
 Vertigo and circulation failure;
 Systemic effects include;
kept in around 6-6.5 [7, 10]. 5 ml of controlled pH
 Metabolic acidosis; sample was taken in a test tube and equal amount of Nash
 CNS depression and coma; reagent was applied. The mixture was kept at 60°C in a
 Respiratory distress; water bath for 10-15min and then cooled down to room
 Renal failure;
temperature. The formaldehyde content was then
 Liver failure;
 Cancer and tumor development; measured using a spectrophotometer.
 Irreversible neurotoxicity
Cancer The Department of Health and Human Services (DHHS)
and the International Agency for Research on Cancer
(IARC) have characterized formaldehyde as a human
IV. RESULTS AND DISCUSSIONS
carcinogen based on studies of inhalation exposure in A. Naturally Occurring Formaldehyde Content in
humans and laboratory animals. Formaldehyde has been
linked to: Fruits and Vegetable Samples
 Nasopharyngeal cancer; Experimental results of naturally occurring
 Gastrointestinal cancer;
formaldehyde contents in different fruit and vegetable
 Possible links to brain cancer and leukemia
samples are presented in Table II and Fig. 1. The
experimental results of banana, grape, apple, pear, carrot,
radish, cucumber and tomato were found compatible with
III. MATERIALS AND METHODOLOGY
the reported values [23]. No reported data of natural
A. Sample Collection occurring formaldehyde were found for pomegranate,
Fruit and vegetable samples were collected from local pomelo fruit, pineapple, ripe papaya, green papaya, and
market and fruit orchards in Dhaka, Bangladesh. Pure lemon. The experimental results provide the baseline data
cow milk sample was collected from a dairy firm in for the above food items.
Mymensingh, Bangladesh. Different UTH (cow) milk B. Formaldehyde Content of Milk Samples
samples (AARONG, MILK-VITA, IGLOO and PRAN),
Table III and Fig. 2 represent the formaldehyde
and powdered (cow) milk samples (DANO, MARKS and
contents found in pure cow milk sample and other UTH
DIPLOMA) were collected from local grocery shops in
and powdered (cow) milk brands. The experimental
Dhaka, Bangladesh.
results for pure cow milk (5.28±3.47 ppm) was
B. Chemicals and Reagents compatible with reported values (3.3 ppm) [23, 24]. The
Reagent grade ammonium acetate, acetic acid, experimental results show that formaldehyde content in
potassium hydroxide, nitric acid (Merck, Germany) UTH milk and powdered milk samples were high
andacetyl acetone (Loba-chemie, India) was used in the compared to that of pure milk (58.79 to 187.75 ppm).The
possible reason for higher formaldehyde content in 50

Formaldehyde content (ppm)

commercial milk samples are dosing of formaldehyde
during milk processing, preservation and/or packaging to 40
improve the shelf life, or, conversion of milk ingredient 30
to primary aldehyde during milk processing [25-27].
20
C. Time Dynamic Behavior of Formaldehyde
formation in Foods 10
The time dynamic behavior of the formation of 0
formaldehyde naturally in foods was investigated in this
study. To understand the time dynamic behavior, the
formaldehyde content and pH of banana samples (AAB
genome of Musa Spp.) were measured for 3 days. Fig. 3
represents the time dynamic behavior of endogenous (b)
formaldehyde content in banana samples (AAB genome
Figure 1. Naturally occurring formaldehyde content of different fruits
of Musa Spp.). and vegetable items; (a) fruits, and (b) vegetables (error bars for n=5
samples)
TABLE II. NATURALLY OCCURRING FORMALDEHYDE CONTENT OF
DIFFERENT FOOD ITEMS (ST. DEV. FOR N=5)
TABLE II I. FORMALDEHYDE CONCENTRATION IN MILK SAMPLES (ST.
DEV. FOR N=5)
Fruit Items Vegetable Items

Formaldehyd Samples Formaldehyde content (ppm)

Samples Formaldehyde Samples
e content
content (ppm) Pure Milk 5.28 ±3.47
(ppm)
Banana (AAA DANO Powder Milk 129.38 ±3.13
genome of Musa 20.68 ±3.17 Carrot 10.86 ±2.05
DIPLOMA Powder Milk 194.08±6.64
Spp.)
Banana (AAB MARKS Powder Milk 90.65±3.54
genome of Musa 14.77±1.08 Radish 6.44 ±2.19 PRAN UTH milk 69.94 ±7.13
Spp.) IGLOO UTH milk 187.75 ±3.11
Grape 15.68 ±6.23 Tomato 14.67 ±6.21
AARONG UTH milk 64.68 ±5.01
Pomegranate 6.65 ±1.05 Cucumber 6.44 ±1.64
Green MILK VITA UTH milk 58.79±6.64
Pomelo fruit 16.33 ±2.24 40.65±5.57
papaya 250
Formaldehyde content (ppm)

Litchi 6.67 ±1.39 Lemon 0 200

Pineapple 20.83 ±3.05
150
Green apple 13.41 ±4.05
100
Red apple 17.18 ±2.69
50
Orange 56.89 ±5.72
0
Mandarin .orange 58.34 ±3.94
Pear 57.69 ±5.47
Mango (Langra) 10.8 ±3.31
Mango(Himsagor
22.4 ±5.64
)
Papaya 55.7±3.03 Figure 2. Formaldehyde content of milk samples (error bars for n=5
samples)

70
Formaldehyde Content, x
Formaldehyde content (ppm)

60 32
50
40 27
30
(ppm)

20
22
10
0
banana(AAB…
banana(AAA…

17
grape

pineapple

orange

mango(langra)
red apple

mandarin
pomelo

pear

papaya
litchi
pomegranate

green apple

mango(himsagor)

12
0 1 Day 2 3

Figure 3. Time dynamic behavior of naturally occurring formaldehyde

content in Banana (error bars for n=5 samples)
(a)
 It was found that the formaldehyde content in banana Formaldehyde is naturally present in food items such as
samples gradually increased with time (Fig. 3). The fruits, vegetables and milk. Therefore, it is important to
possible reason of the gradual increase of formaldehyde know the concentration of naturally occurring
content is the formation of S-adenosyl-L-methionine formaldehyde in food items to determine any external
(SAM) during banana ripening, which is associated with formaldehyde dosage in food samples. This study offers
endogenous formaldehyde production [17, 28]. Banana is baseline data of formaldehyde content naturally found in
a climacteric fruit, which is associated the with increase different food items. In this study, the naturally occurring
in respiration and ethylene production during the ripening formaldehyde content of fifteen fruit samples, six
process [29]. SAM, a major methyl donor in cells, is vegetable samples and eight milk samples were analyzed.
associated with the biosynthesis of ethylene [30, 31]; it According to the European Food Safety Authority (EFSA)
has been reported that, during ripening process, the SAM the daily exposure to formaldehyde from food should not
level increases in climacteric fruits [30]. In addition, the exceed 100 mg/kg (100 ppm) food per day [6].
pH value of banana sample changes from 4.5 (t = 0) to Formaldehyde content of fruit samples ranged between 6
5.2 (t = 3 days) during the ripening process, which to 58 ppm; for vegetables the range was in between 0
indicates a decrease in acid content [32]. Formaldehyde (zero) to 41 ppm; and for pure cow milk sample the
formation from methylated compounds can be formaldehyde concentration was about 5 ppm. These
represented by the following reaction (equation 1) [33]: values are in accordance with other published results, and
less than the maximum daily exposure set by EFSA [6,
𝐷𝑒𝑚𝑒𝑡ℎ𝑦𝑙𝑎𝑠𝑒 23]. However, the formaldehyde content of the seven
𝑀𝑒𝑡ℎ𝑦𝑙𝑎𝑡𝑒𝑑 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑𝑠 → 𝐹𝑜𝑟𝑚𝑎𝑙𝑑𝑒ℎ𝑦𝑑𝑒 commercial (cow) milk samples were found higher than
(1) that of pure (cow) milk sample; in three commercial milk
samples the formaldehyde level exceeded the daily
t=0 a 0 exposure limit set by EFSA. Addition and/or formation of
formaldehyde during milk processing and preservation
t=t a-x x are the possible reasons to have high formaldehyde
concentrations in commercially available milk samples.
The order of the kinetics of the above reaction Further study is required to analyze the possible reasons
(Equation 1) can be described as [33]: to have high formaldehyde concentration in commercial
milk samples and associated health effects. In addition,
First order: 2.303×log (a-x) = - Kt + c (2)
the time dynamic behavior of the formation of
Second order: 1/(a-x) = Kt + c (3) endogenous formaldehyde in banana samples was
analyzed. The reaction kinetics showed that the
The experimental results of Fig. 3 were plotted endogenous formaldehyde production in banana sample
following the kinetic equations (Equations 2 and 3); it follows second order reaction kinetics, and is likely to be
was found that the endogenous formaldehyde formation related to the concentration of methylated compounds
in banana followed second order kinetics (Equation 3) present in the banana sample. Methylated compounds
with a rate constant (K) 0.2332 mole-1day-1 (per mole of such as S-adenosyl-L-methionine (SAM) play an
methylated compound per day) (Fig. 4). important role in many biochemical reactions in plants,
0.6
such as: the biosynthesis of polyamines and plant
y = 0.2332x + 0.0826 hormone ethylene [30, 31]. However, the dynamics and
0.5 R² = 0.9943 the key factors of the formation of formaldehyde in other
food items could be different depending on the molecular
0.4
structure and aging pattern of the food items.
0.3
1/(a-x)

ACKNOWLEDGEMENT
0.2
This research was supported by BCEF Academic
0.1 Research Fund, CASR Research Fund, and financial
0
support from the Department of Chemical Engineering,
0 1 Day 2 3 BUET. The authors would also like to thank Ms. Shawly
Samira and Mr. Mursalin Rahman for their technical
Figure 4. Kinetics of naturally occurring formaldehyde formation in assistance.
Banana (error bars for n=5 samples)
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 Farrhin Nowshad is pursuing her B.Sc. in
Chemical Engineering from Bangladesh
University of Engineering and Technology.
She is also working as an undergraduate
researcher in Bio and Environmental
Technology (BET) group, under the
supervision of Dr. Mohidus S. Khan,
Assistant Professor, Department of Chemical
Engineering (BUET). Her research interest
includes biochemical engineering,
engineering properties of food materials and food chemistry.

Md N. Islam graduated in Chemical

Engineering, from Bangladesh University
of Engineering and Technology (BUET) in
2015. Currently he is pursuing his M.Sc. in
Chemical Engineering and also working as
a researcher in Bio and Environmental
Technology (BET) group under the
supervision of Dr. Mohidus S. Khan. His
research interest includes, food chemistry
and paper based point-of-care detection
devices.

Dr. Mohidus S. Khan is an Assistant

Professor at the Department of Chemical
Engineering, Bangladesh University of
Engineering and Technology (BUET). Dr.
Khan completed his BSc in Chemical
Engineering (2004) Bangladesh University of
Engineering and Technology (BUET).
Starting in 2006, Dr. Khan completed his
PhD in 2010 in Bio-surface and
Biotechnology from Monash University,
Australia. Dr. Khan worked as a Post-
Doctoral Fellow at the Department of Chemistry, McGill University,
Canada to continue his research work on biotechnology (2010-13).
Since 2013, Dr. Khan is working as an Assistant Professor in the
Department of Chemical Engineering, BUET. He also served as a
Visiting Professor (2015) at McGill University, Canada, and as a
Visiting Research Scholar (2016-17) at Texas A&M University, USA.
Dr. Khan has been involved in food, environment, and biotechnology
related projects led by the Government (GoB), international donor
agencies, and local Industries.
He is an aspiring researcher and innovator. His research interests
encompass Food, Biotechnology and Environmental Engineering. To
date, he has authored and co-authored over 50 technical articles, which
include peer-reviewed journal and conference articles, international
patents, industrial reports, books and book chapters. He also serves as a
reviewer of several reputed international journals.

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