1

Introduction
Acrylamide or acrylic amide, a carcinogenic compound with a chemical formula
C3H5NO. The International Agency for the Research on Cancer has classified
acrylamide in Group 2A carcinogens. Acrylamide has not only been demonstrated as a
carcinogen but also exhibit neurotoxic effects. Other negative health risks associated
with acrylamide include reproductive toxicity, genotoxicity, tumors of lungs, uterus,
mammary gland etc. Historically, acrylamide was only present in water and its threat
to humans was not a major concern. However, the modern discovery of acrylamide
formation in certain fried, processed and baked foods with high levels ranging up to 10
mg/kg were found in highly consumed food products [1].

Asparagine, a nonessential amino acid, in combination with various carbonyl

compounds at temperatures greater than 120°C, has been identified as the amino acid
precursor for acrylamide formation in food as a result of browning procedure called
Millard reaction. It can also be formed from acrylic acid or acrolein, resulted from
sugars and glycerols [2]. Potato products, such as French fries and chips are among the
food items carrying the highest contents of acrylamide and binds to haemoglobin to
form an adduct [3].

Acrylamide formation is dependent on the quantity of reducing sugars (asparagine),

cooking processes, temperature, time, pH and surface to volume ratio of food materials.
These factors were taken into consideration through several effective measures that
include raw materials modification, optimization of heat processing parameters,
numerous compounds addition, pH modifiers and proteins or amino acids blanching
[4].

Acrylamide (AA) can be ingested, inhaled or absorbed through the skin and due to its
solubility, it is distributed throughout all body tissues, through passage in the blood and
is majorly found in the liver, brain, heart, kidneys, breast milk and placenta. In human
body, acrylamide is oxidized by cytochrome P450 and forms glycidamide (metabolite),
which has higher binding affinity for serum, DNA, hemoglobin and enzymes (in vivo)
to produce neurological damage and mutagen formation. Signs of acrylamide contact
constitutes weakness in muscles (skeletal) and ataxia [5].
In humans, when acrylamide gets in contact with skin it distributes itself to all tissues
and forms glycidamide, (reactive epoxide). It makes adducts with DNA and proteins
 2

leading to genotoxicity and mutagenicity. The other products of maillard reaction tend
to increase or decrease acrylamide toxicity [5].

Acrylamide concentration significantly reduces after the gastrointestinal digestion. As

food swallowing causes a change in structure and composition by chewing, the change
in pH occurs along with, dilution and different enzymes present in mouth, stomach and
intestine act on to food It is important to identify what measures may be taken to
decrease both the production and effects of acrylamide in foods. Acrylamide causes
cancer and neurotoxic effects therefore its development in foods and the effect of this
subjection on human health needs to be particularly understood. It is also necessary that
the method for determining acrylamide in foods is not only applicable to a wide range
of foods, but also adequately strong for analyzing large numbers of samples [6].

Through literature study it has been found that acrylamide is carcinogenic in nature and
its detection in food has been found with various techniques such as liquid
chromatography, high performance liquid chromatography with an ion exclusion
column and UV detection, FTIR, Gas Chromatography coupled to Mass Spectrometry,
portable vibrational spectrometers, Gas chromatography coupled to an electron capture
detector, nitrogen phosphorus detector, flame photometric detection and flame
ionization [7]. Present work will evaluate quantity of AA in food through high
performance liquid chromatography (HPLC) [8].
 3

Aims & objectives

As human body eats a lot of processed baked and cooked foods, exposure of acrylamide
is maximum. If a greater amount of this carcinogen is found then, appropriate measures
will be taken to reduce such foods in our diet so, cancers and mutations be prevented.
The objectives will include:

• To evaluate acrylamide concentration in human blood and different foods.

• To develop a biosensor for correctly detecting acrylamide in blood.

• To study the factors involved in the development of adduct of haemoglobin-
acrylamide.
 4

Review of Literature
Yadav et al. (2018) described the construction of an electrochemical paper based
device (EPAD) for detection of acrylamide, a carcinogen and neurotoxin generated
during thermal processing of foods. Haemoglobin nanoparticles were prepared by
desolvation method and characterized was done. The HbNPs were drop casted onto
PAD for electrochemical detection of acrylamide. The EPAD showed the optimum
response within the working range of sensor (0.1 nMe100 mM) with 0.1 nM LOD.
Analytical recovery, coefficient of variation was evaluated. The EPAD was employed
for detection of acrylamide in various processed foods [9]
Mesias et al. (2018) performed liquid chromatography-electrospray ionization tandem
mass spectrometry technique to determine acrylamide content in French fries.
Acrylamide, reducing sugar, moisture, color and polar compounds were evaluated.
Results showed that with increasing degree of automatization and control of frying
process acrylamide concentration can be reduced [10].

Norouzi et al. (2018) examined acrylamide levels in different bread samples using
highly sensitive ultrasonic assisted extraction and micro-extraction method.
Acrylamide concentration, percentage recovery, relative standard deviation, analysis of
variance, limits of quantification and detection were evaluated. Results have shown that
this method is an accurate, sensitive, fast and reliable sample-pretreatment method for
determining the very low amount of acrylamide in various bread samples [11].

Nguyen et al. (2017) investigated acrylamide and 5-hydroxymethylfurfural (HMF)

formation during biscuit baking. Effects of ratio of reducing sugars and aspargine,
acrylamide level and reaction rate constants were evaluated. Results showed no clear
correlation between acrylamide and HMF in baked biscuits, nor between asparagine
and sum of glucose and fructose amounts in wheat flour [12].

Veni et al. (2014) performed high-performance liquid chromatography for the

estimation of acrylamide in potato chips. Concentration of acrylamide, peak area,
internal standard peak area and response factor was assessed from the calibration curve
(Linear) of acrylamide. Mean, precision, limits of quantification and detection and
relative standard deviation were also investigated. Results showed that this method was
used only for estimating higher amount of acrylamide in potato chips [8].
 5

Lim et al. (2014) proposed a simple, unique method for determining the amount of
acrylamide in potato chips, French fries, and coffee, after derivatization with d cysteine
using liquid chromatography coupled to tandem mass spectrometry. Parameters such as
acrylamide level, mean, precision, accuracy, standard deviation and limits of
quantification and detection were examined. Results showed that proposed method
offered easier manipulation of samples, shorter analysis duration and opportunities for
acrylamide analysis in case of small sample sizes [13].
Al-Janabi et al. (2013) quantified acrylamide level in different variety of chips and an
Iraqi meal (Harissa). Acrylamide level, spiked recoveries, relative standard deviation
and limits of quantification and detection were assessed. Results showed that this
method is simple, benign, sustainable and used minimal amounts of reagents which
avoided production of toxic residues [14].

Geng et al. (2011) evaluated acrylamide content in starch-based foods by HPLC with
pre-column ultraviolet derivatization. Acrylamide level, mean, precision, accuracy,
standard deviation, spiked recovery and limits of quantification and detection were
evaluated. Results showed that this method is a new, low-cost, and robust alternative
for conventional investigation of acrylamide [15].

Khoshnam et al. (2010) reported a new sensitive method for the analysis of acrylamide
in different potato chips using HPLC via acetone extraction. Recovery test at different
spike levels for two different samples in different days were calculated which
comprised mean, limit of quantification, limit of detection, relative standard deviation
and standard deviation. Results showed that the proposed method could successfully be
applied for the routine analysis of AA in potato chips due to its low cost instrumentation
and the use of fewer chemicals [16].

Zhang et al. (2007) determined acrylamide content in potato crisps by using

ultraperformance liquid chromatography coupled to electrospray ionization tandem
mass spectrometry technique. Spiked recoveries and relative standard deviation were
calculated from detected level of acrylamide. Results showed that the present
quantitative method could be applied for rapid determination of acrylamide in many
investigations [17].
 6

Materials and Method

Instrumentation
• Potentiostat/Galvanostat

Plan of Work

SAMPLE COLLECTION

PREPARATION OF HBNPS AND FOOD

SAMPLES

CHARACTERIZATION OF HBNPS

PREPARTION OF PAPER BASED

ELECTROCHEMICAL BIOSENSOR

FABRICATION OF BIOSENSOR FOR

ACRYLAMIDE SENSING

CYCLIC VOLTAMMOGRAMS ARE OBTAINED

FROM GALVANOSTAT

APPLICATION OF ACRYLAMIDE ONTO THE

BIOSENSOR

EVALUATION OF ACRYLAMIDE LEVEL FROM

VOLTAMMOGRAMS
 7

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LC−MS MS or GC−MS analysis of Acrylamides in Various food matrices. J.

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 8

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