ENV460 Course File

15/12/2020

ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Introduction

LESSON 7: PRE-CONCENTRATION • Pre-concentration - the reduction of a larger sample
USING SOLVENT EVAPORATION into a smaller sample size.

AHMAD RAZALI ISHAK • Most commonly carried out by using solvent
evaporation procedures after an extraction technique.
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY
FACULTY OF HEALTH SCIENCES • The most common approaches for solvent evaporation
are Rotary evaporator, Centrifugal evaporator,
UiTM PUNCAK ALAM, 42300 KUALA SELANGOR Kuderna-Danish Evaporative concentration, Nitrogen
blow down evaporator.

• the evaporation method is time consuming, with a high
risk of contamination from the solvent, glassware and
blow-down gas.

14

Rotary Evaporation Rotary Evaporation -Disadvantages

• used for evaporating single volatile • Since the glassware is under high vacuum, there’s
samples at relatively larger sample always a risk of the solvent superheating and boiling
volumes. over or “bumping.”

• A round bottom flask is used where it • Once a sample boils over, there is a high risk of cross-
is lowered in a heated water bath. contamination and losing material.

• The flask is rotated to increase the • Improper selection or defective glass could lead to an
surface area and provide an even implosion.
transfer of heat.
• Evaporating multiple samples must be done in series,
• The sample is placed under vacuum which can be time consuming.
to lower the boiling point of the
solvent. 15 16

Centrifugal evaporator Centrifugal evaporator - Disadvantages

• used to evaporate a large • Not suitable for higher boiling point solvents like DMSO
number of samples at once since or water, as it can be time-consuming, typically
they can accommodate multiple overnight.
centrifuge tubes at a time.
• the samples are well balanced and require proper
• Works under high vacuum to centrifuge tubes
lower the boiling point.
• Improper glassware tends to break under centrifugal
• Samples are placed in a force
multiplexed heated compartment
where they are centrifuged to 17 18
avoid bumping.

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Kuderna Danish Kuderna Danish
Evaporative concentration Evaporative concentration

• Consists of an evaporation flask • Solvent vapors rise and condense within a narrow
connected to a Snyder column and opening Snyder column.
to a concentrator tube.
• Sufficient pressure needs to be generated by the
• The sample is placed in the solvent vapors to force their way through the column.
apparatus and heated with a water
bath (15–20◦C above the boiling • Initially, a large amount of condensation of these
point of the organic solvent). vapors returns to the bottom of the K–D apparatus.

• Concentrator tube should be • wash the organics from the sides of the evaporation
partially immersed in the water bath flask and assists in the process of re-condensing volatile
and all the entire lower part of the organics.
evaporation flask to be bathed with
hot vapor. • This process of solvent distillation concentrates the
sample to approximately 1–3 ml in 10–20 min.

• Escaping solvent vapors are recovered by using a
condenser and collection device.

19 20

Nitrogen/Gas Blow-Down References and Credit

• A gentle stream of (high-purity) purge gas is passed • Dean, J. R. (2003). Methods for environmental trace
over the surface of the sample. analysis (Vol. 12). John Wiley and Sons.

• the purge gas is directed towards the side of the • https://biochromato.com/blog/types-of-solvent-
vessel, and not directly onto the top of the extract to evaporators/
induce a swirling action.

• The extract-containing vessel may be partially
immersed in a water bath to speed up the
evaporation process.

21 22

Thank you.

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 7: ATOMIC ABSORPTION At the end of this lesson, student should be able to:
SPECTROSCOPY (AAS) • Describe the principle of AAS
• Explain the component of AAS
AHMAD RAZALI ISHAK • Describe the advantages of disadvantages of FAAS

CENTRE OF ENVIRONMENTAL HEALTH & SAFETY and GFAAS
FACULTY OF HEALTH SCIENCES • Explain several applications of AAS

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR 2

Introduction Atomic Absorption Spectroscopy (AAS)

• Various method for inorganic compounds -atomic • AAS is an instrumental
spectroscopy, X-ray fluorescence spectroscopy, mass technique for determining
spectrometry, electrochemical approaches and the concentration of a
chromatography. particular metal element in
a sample.
• The purpose of this present section is based on atomic
spectroscopy -atomic absorption. • AAS can be used to analyze
the concentration of over
3 62 different metals in a
solution.

4

Principle of AAS Principle of AAS

• The metal vapor absorbs energy from an external light • During excitation from ground state to excited state,
source, and the electrons jump from the ground to the the atom absorbs light of the same characteristic
excited states wavelengths.

5 • The same thing happen when returning from the
excited state to the ground state.

• The absorbed light (absorbance), can be used as a
measuring signals known as spectrum.

• The intensity of the absorbed light (absorbance) is
proportional to the concentration of the element in
the sample.

• A calibration curve can thus be constructed
[Concentration (ppm) vs. Absorbance]

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AAS Components - FAAS AAS Components - GFAAS

HCL/ EDL lamp Atomizer -Flame Photomultiplier HCL/EDL Lamp Photomultiplier
(Contains element to be analyzed) Containing the (Contains element to be analyzed)
atomized sample
Air
Acetylene Monochrometer Atomizer – Monochrometer
(Select wavelength) Graphite Furnace (Select wavelength)
(Electrically heated)
Nebulizer

sample

7 8

1. Light source/ Lamp Cont.

Hollow cathode lamp Electrodeless discharge lamps

• 2 types of light sources are mostly • EDL contain the element
used Hollow-cathode lamps (HCL) in a small quartz tube
and electrodeless discharge lamp filled with a noble gas.
(EDL)
• A high-frequency field
• HCL contain a cathode of the leads to a plasma within
analyte element and an anode, the tube, in which the
and are filled with a noble gas. element is excited and
emits specific light.
• When a high voltage is applied
across the anode and cathode, the 9
metal atoms in the cathode are
excited into producing light with a
certain emission spectrum.

10

Cont. 2. Atomizer

• The type of HCL/EDL depends on the metal • Atomizer/Atomization cell -the
being analyzed. site were the sample is
introduced
• For analyzing the concentration of Cu in an ore,
a Cu lamp would be used, and likewise for any • Flame or graphite-furnace
other metal being analyzed. • Causes the metal-containing

• The electrons of the atoms in the sample to be dissociated and
flame/graphite can be promoted to higher liberated from a hot
orbitals for an instant by absorbing a set environment.
quantity of energy (a quantum). • Such an environment of the
atomization cell is sufficient to
11 cause a broadening of the
absorption line of the metal.

12

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Atomization in Flame AAS Cont.

• Three steps are involved in turning a liquid sample • The flame is arranged laterally long (usually 10 cm)
into an atomic gas in flame AAS: and not deep.
• Desolvation – the liquid solvent is evaporated,
and the dry sample remains. • The height of the flame must also be monitored by
• Vaporization – the solid sample vaporizes to a controlling the flow of the fuel mixture.
gas
• Volatilization – the compounds making up the • A beam of light passes through this flame at its
sample are broken into free atoms. longest axis (the lateral axis) and hits a detector.

13 14

Nebulizer FAAS

• Sucks up the liquid sample (= aspiration) Disadvantages
• creates a fine aerosol for introduction into flame • Only 5-15% of nebulized
• Mixes aerosol, fuel and oxidant thoroughly, creates a
sample reached the flame
heterogeneous mixture • Minimum sample volume
• The smaller the size of the droplets produced, the higher
of 0.5-1.0 mL is needed to
the element sensitivity give a reliable result.
• Fuel = Acetylene, Oxidant = Compress air/acetylene • Sample which are viscous
• Mixture of Acetylene/Air (2300 ˚C), require dilution with
• Mixture of Acetylene/Nitrous oxide(2700˚C) solvent.

15 16

GFAAS GFAAS

• The sample is holder is graphite Advantages
tube • Small sample size
• Very little or no sample preparation is need
• The sample is placed in the • Sensitivity is enhanced
graphite furnace and electrically • Direct analysis of solution, slurries and solid samples
heated
Disadvantages
• An electro thermal atomizer system • Analyte maybe lost at the ashing stage
can produce temperatures as high • The sample may not completely atomized
as 3000°C. • Analytical range is relatively low

• Beam of light pass through the tube 17 18
• 3 stages

• Drying the sample
• Ashing of organic matter
• Vaporization of analyte atom

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Monochromator Detector

• Use to select a specific wavelength of light • The light select by monochromator is
which is absorbed by the sample and to exclude directed onto as detector that is typically a
other wavelength photomultiplier tube (PMT).

• The selection of specific light allow the • PMT –Convert the light signal into an electric
determination of specific element signal proportion to intensity.

• The processing of electric signal is fulfilled by
a signal amplifier.

• The signal could be display for readout or
printed by the requested format.

19 20

Calibration curve Calibration curve

• A calibration curve is used to determine the 22
unknown concentration of an element in a
solution

• The instrument is calibrated using several solution
of known concentrations

• The absorbance of each known solution is
measured and then a calibration curve of
concentration VS absorbance is plotted.

• The unkown concentration is calculated from
calibration curve.

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Examples AAS Applications

• Lead is extracted from a sample of blood gave an absorbance of • Water analysis
0.340 in an AAS. Using the following calibration curve, find the • Food analysis
• Analysis of animal feedstuffs
concentration of lead ions in the blood sample. • Analysis of additives in lubricating oils and

[Pb+2] (ppm) Absorbance Calculated Pb (II) concentraions (ppm) Absorbance greases
• Analysis of soils
0.000 0.000 0.357 0.340 • Clinical analysis (blood samples: whole blood,

0.100 0.116 • Used the equation to plasma, serum)
0.200 0.216
0.300 0.310 calculate the concentration 24
0.400 0.425 of Pb (II) ions with an
0.500 0.520 absorbance of 0.340.

Lead (II) Calibration Curve • The result, 0.357 ppm, is
displayed above the graph.
0.600

0.500 y = 1.0505x
0.400 R2 = 0.9988

Absorbance 0.300

0.200

0.100

0.000 0.100 0.200 0.300 0.400 0.500 0.600
0.000

[Pb+2] (ppm)

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Thank you.

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 8: High Performance Liquid At the end of this lesson, student should be able to:
Chromatography (HPLC)
• Describe the fundamental of HPLC theory
AHMAD RAZALI ISHAK • State different type of chromatography
• Explain the separation modes of HPLC (Normal and
CENTRE OF ENVIRONMENTAL HEALTH & SAFETY
FACULTY OF HEALTH SCIENCES reverse phase)
• State different type of HPLC detectors
UiTM PUNCAK ALAM, 42300 KUALA SELANGOR • Explain HPLC systems (Isocratic and gradient)
• Describe two type of calibration method in HPLC

analysis

2

Fundamental of HPLC – Basic Theory Fundamental of HPLC – Basic Theory

What is chromatography? How can the separation be carried out?
• Chromatography is one of the separation • Separation can be carried out by the column.
technique.
• The main purpose of chromatography is to
separate and quantify the target sample in the
matrix.

34

Fundamental of HPLC – Basic Theory Fundamental of HPLC – Basic Theory

Interaction Separation can be done by the column
between packing
material and 6
sample
• Due to difference

interaction
between packing
material and
sample,
separation can
be done.

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What kind of chromatography? Advantage of Chromatography

• High Performance Liquid chromatography (HPLC) • Simultaneous Analysis
• Gas Chromatography (GC) • High Resolution
• Thin-Layer Chromatography (TLC) • High Sensitivity (ppm-ppb)
• Capillary Electrophoresis(CE) • Small Injection Volume (1-100uL)

78

Advantage of HPLC Separation modes of HPLC

• Moderate analysis condition -no need to vaporize the • Normal Phase mode
sample like GC • Reversed Phase mode
• Reversed Phase Ion Pairing mode
• Easy to fractionate the sample and purify • Chiral separation mode
• Good repeatability (C.V. < 1%) • Ion Exchange mode
• SEC mode ( GPC / GFC )

9 10

Normal Phase Mode Normal Phase Mode

M. Tswett : • First developer used
First developer of  CaCO3 for Separation Column
chromatography  Petroleum Ether for Developed Solvent

Ber. Deut. Botan. Ges., 24, • We defined this combination as Normal Phase
384 (1906) mode
Adsorptionsanalyse und
chromatographiische • Column : polar property
Methode. Anwendung auf • Solvent : non polar property
die Chemie des
Chlorophylls. 11 12

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Normal Phase HPLC Columns What is the interaction?

• Silica gel type : general use

• Cyano type : general use
• Amino type : for sugar analysis

• Diol type : for protein analysis

13 14

Hydrogen bonding Selection of solvents for mobile phase of
Normal Phase mode
• If the sample has:
 -COOH : Carboxyl group Primary solvents
 -NH2 : Amino group • Hydrocarbons (Pentane, Hexane, Heptane, Octane)
 -OH : Hydroxyl group • Aromatic Hydrocarbons (Benzene, Toluene, Xylene)
• Methylene chloride
• “Hydrogen Bonding” becomes strong. • Chloroform
• Carbon tetrachloride
• If the sample does not have any functional
groups such as hydrocarbon or Secondary solvents
• Alcohols (2-propanol, ethanol, methanol), Methyl-t-butyl ether
• If the sample has balk group, due to stereo
hindrance (MTBE),Diethyl ether, Tetrahydrofuran (THF), Dioxane, Pyridine, Ethyl
acetate, Acetonitrile, Acetone.
• “Hydrogen Bonding” becomes weak. • Select one of primary solvents and add secondary solvents in
order to adjust retention time. Solvents which possess UV
15 transparency may be better for easy detection.

16

Increase of solvent polarity Reversed Phase Mode

• Column : Non-polar property
• Solvent : Polar property

- Normal Phase mode -

• Column : polar property

• Solvent : non polar property

17 18

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Reversed Phase HPLC Columns Whatt is the interaction?

• C18 (ODS) type
• C8 (octyl) type
• C4 (butyl) type
• Phenyl type
• TMS type

Hydrophobicity 19 20

Selection of solvents for mobile phase of
Reversed Phase Mode

• Water (buffer) + Organic solvents
 In case of buffer, buffer concentration and
pH is important
 As a organic solvent, methanol, acetonitrile
or THF are very commonly used.

• Optimization of water (buffer) and organic
solvents’ ratio is very important.

21 22

Increase of solvent polarity Normal vs Reversed Phase

Normal Phase
• Good separation for stereo isomer such as
Vitamin E etc.
• Changeable Retention Time (RT)

Reversed Phase
• Good repeatable retention time
• Rugged stationary phase

23 24

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Instrumentation – HPLC components HPLC Detectors

• Ultraviolet / Visible detector (UV/VIS)
• Photodiode Array detector (PDA)
• Electrochemical detector (ECD)
• Refractive Index detector (RID)
• Fluorescence detector (RF)
• Conductivity detector (CDD)
• Mass spectrometer detector (MS)

25 26

HPLC Systems Isocratic elution system

Isocratic elution system - 27 28
Single solvent of constant
composition

Gradient elution system -
Multi solvents of
changeable composition
• High pressure gradient

system
• Low pressure gradient

system

Gradient elution system Isocratic VS Gradient

Binary (High Pressure Gradient) 29
• Two solvents of changeable

composition
Ternary (High Pressure
Gradient)
• Three solvents of

changeable composition
Quaternary (Low Pressure
Gradient)
• Four solvents of changeable

composition

30

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Calibration methods 15/12/2020

• External calibration method Calibration methods

• Internal calibration method

31 32

Calculations Calibrations

External calibration External standard calibration
• Separation is not difficult
Internal calibration • Injection error will directly influence the
33 quantitative result

Internal standard calibration
• Injection error can be eliminated
• Recovery in the pre-treatment procedure can
be estimated
• Separation is slightly difficult
• Difficult to look for the IS compound

34

Thank you.

6

Chromatography 15/12/2020

The stationary phase is a liquid Gas Chromatography
dispersed on a solid support
whereby the separation is Ahmad Razali Ishak (PhD)
achieved by partition [email protected]

The active solid (e.g. charcoal,

molecular sieves) whereby the
separation is achieAvdesodrpbtioyn
adsorption
HPLC

Refer to Table 30-4 (page 921)
Fundamentals of Analytical

Chemistry

• MUST know the basic components of What is it ? • Technique
gas chromatography and their • Analytical chemistry
functions. • Separation and Analysis
• Volatile compounds
• HAVE to know the flow analysis in gas
chromatography.

• SHOULD understand the principle of
mobile phase and stationary phase in
gas chromatography.

• MUST be able to explain the
chromatogram.

• IMPORTANT to differentiate between
liquid chromatography and gas
chromatography.

Keyword GC sample is liquid
TEMPERATURE
• Vaporization process
• Liquid change to gas

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Set up of gas chromatography 5
2

4
3

STATIONARY PHASE

1

MOBILE PHASE

Carrier gas Injector

• Mobile phase – a carrier gas, gas • In the vaporization chamber, heated 50°C above the lowest
flow through the system and boiling point of the sample and mixed with carrier gas to
carries the injection sample. transport the sample to the column.

• Properties of carrier gas – must
be dry (no water), free of
oxygen, constant flow rate and
chemically inert.

• E.g. Helium - large range of flow
rate, compatible with many
detectors.

• E.g. Nitrogen, Argon, Hydrogen,
Carbon Dioxide.

Column and Oven Packed column

• Stationary phase – oven and colum GasGclahsrsocmoatluomgrnaphy
• Thermostated oven control the temperature of the column column
• Open tubular column / capillary column
• Packed column

Capillary colum

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Packed column Capillary column Packed column

A finely divided, inert,
solid support material
(commonly based on
diatomaceous earth)
coated with l iquid
stationary phase.

M ost packed col umns are
1.5 - 10m in length and
have an internal
diameter of 2 –4mm.

Capillary columns have an internal diameter of a few tenths of a Detector
millimeter. The inner walls are coated with thin layer of
stationary phase. • Quantitative measurement of
the sample
WCOT, Wall Coated Open Tubular
SCOT, Support Coated Open Tubular • MS
PLOT, Porous Layer Open Tubular (Gas-solid chromatography) • FID
FSOT, Fused Silica Open Tubular • TCD
• ECD
Flame Ionisation Detector (FID) • Generate the signal and covert

Compounds are burnt in the flame to chromatogram
producing ions and thus causing an
increase in current flow between 3
the jet and collector.
Detects carbon/hydrogen
containing compounds.
Wide range of l inear response

15/12/2020

detector Chromatogram

1. Adequate sensitivity 10-8 to 10-15 g sol ute/s Rt A
2. Good stability and reproducibility
3. A linear response-extends over several orders of magnitude Rt B
Rt = Retention time
4. A temperature range fr room temp. to 400°C
5. A short response time-independent of flow rate
6. High reliability and ease of use
7. High predictable and selective response
8. Nondestructive of sample

A mixture of cyclohexane and dioxane is eluted on a non-polar gas Differentiate between packed column and open tubular
chromatography column. The chromatogram obtained is shown column ..
below:

cyclohexane

1,4 - dioxane

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Liquid Chromatography Gas Chromatography

Mobile phase is a liquid Mobile phase is a gas

Separation is based on interaction of solute with Separation is primarily based on the boiling points

the chromatography medium of solute molecules

Can be performed in a sheet or a column Can be carried out only in a column

Can be used to separate any soluble Can be applied in the separation of volatile
compound, e.g. amino acids, proteins, drugs, compounds and gaseous mixtures

nucleic acids, lipids, antioxidants, carbohydrates,
and natural and artificial polymers

Usually carried out at room temperature so heat- Performed at higher temperatures so thermally
sensitive compounds can be safely analyzed using labile substances might get denatured

the technique

Refer to Table 32-3 (page 993) Solute retention here is based on the interaction of Separation is based on the boiling points of the
Fundamentals of Analytical Chemistry
solutes with the mobile and stationary phases so it solute molecules so it is not very flexible in terms of

is easy to optimize results optimizing separation

This is a relatively slower technique The analysis is faster and usually measured in
minutes, although it can take as little as a couple

of seconds

Usually gives a greater peak or broader band Provides comparatively better resolution
resulting in lower resolution

Uses polar solvents like water or methanol Uses any solvent that vaporizes

Which of the following is not used for detection in GC?
A. Mass spectrometry
B. NMR
C. Flame ionisation
D. Electrical conductivity

The GC trace obtained after an experiment is called a
A. chromatograph
B. chromatogram
C. chromatophore
D. graph

What is the typical internal diameter of fused silica capillary The column is heated to
columns? A. prevent analyte condensation within the column
A. 0.2-0.3 mm B. control elution of the different analytes
B. 0.3-0.5mm C. reduce band broadening to get sharper peaks
C. 0.5-1.0 mm D. all of these
D. 1.0-2.0 mm
Which of the following gases is unsuitable for use as a GC
Which of the statements is correct? carrier gas?
A. Gas chromatography is used to analyse gases A. Nitrogen
B. Gas chromatography is used to analyse solids B. Helium
C. Gas chromatography is used to analyse gases, C. Oxygen
solutions and solids D. All of the above
D. All of the above

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ENV460 ANALYTICAL TECHNIQUE & INSTRUMENTATION Learning outcomes

LESSON 10: ANALYTICAL METHOD At the end of this lesson, student should be able to:
VALIDATION
• Define method validation
• Describe the necessary of method validation
• Explain the analytical characteristics used in method

validation

AHMAD RAZALI ISHAK

CENTRE OF ENVIRONMENTAL HEALTH & SAFETY
FACULTY OF HEALTH SCIENCES

UiTM PUNCAK ALAM, 42300 KUALA SELANGOR

2

Definition Why is Method Validation Necessary?

• Validation of an analytical method is the process • To prove what we claim is true
that establishes, by laboratory studies, that the • To minimize analytical and instrumental errors
performance characteristics of the method meet the • To increase trust of laboratory customers
requirements for the intended analytical applications • To ensure the quality of the test results
• To meet accreditation requirement

34

Selecting Analytical Methods When should Methods be Validated?

• Priority - Standard method, Official Methods that • New method development
have been studied. • Revision of established methods
• When established methods are used in different
• In the absence of standard/official method, use
another method (in-house method) but must first laboratories/different analysts etc.
validate. • Comparison of methods

56

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When should Methods undergo How should methods be validated?
Verification?
• Validation in a single laboratory:
• Major instruments are replaced • Comparisons with certified reference materials
• Methods are used for the first time by a new staff (CRMs)
• The laboratory takes an already validated and • Comparisons with other methods that are
validated.
verified method into use after it has not been used
for a long time • Validation in a group of laboratories:
• Inter-laboratory comparisons

78

Analytical Characteristics Used In Method Linearity
Validation
• Linearity of an analytical
• Linearity procedure as the ability to
• Selectivity/Specificity obtain test results of
• Limit of Detection variable data which are
• Limit of Quantitation directly proportional to the
• Precision concentration of analyte in
• Recovery the sample.
• Robustness
• Provides a plot of
9 measurement response on
the y-axis versus on the x-
axis.

• Linear regression equation
• R2 = Correlation Coefficient

10

Selectivity/Specificity Limit of Detection (LOD)

• The ability of the method that can determine a • The lowest concentration of analyte in a sample that
particular compound in the matrices without can be detected, but not necessarily quantitated
interference from matrix components (e.g., under the stated conditions of the test.
impurities, degradation products and matrix • Analyze 10 independent sample blanks
components). measured once each
• spiking samples • Calculate standard deviation (s) of sample
blank
11 • Calculate mean value of sample blank

• LOD = x + 3 SD (x = average blank sample
reading)

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Limit Of Quantitation (LOQ) Precision

• LOQ is the lowest concentration of an analyte that • The closeness of agreement between independent
can be determined with acceptable precision test results obtained under stipulated conditions.
(repeatability) and accuracy under the stated (How close results are to one another).
conditions of the test.
• Analyze 10 independent sample blanks 14
measured once
• Calculate standard deviation (s) of sample
blank
• Calculate mean value of sample blank
• LOQ = x + 10 SD (x = average blank sample
reading)

13

Definition Cont..

• A: Accurate and precise. The darts are average Repeatability
position is the center. • Precision under repeatability conditions, i.e.

• B: Precise, but not accurate. The darts are together conditions where independent test results are
but not hit the intended mark. obtained with the same method on identical
test items in the same laboratory by the same
• C: Accuracy and precision are poor. The darts are operator using the same equipment within short
not together and not near the center. interval of times.

15 Intermediate precision (internal reproducibility)
• Precision measured between analyses, over

extended timescale, within a single laboratory.

16

Cont.. Recovery

Reproducibility: • Recovery is the ratio of the observed value to the
• Precision under reproducibility conditions, i.e. expected value. Recovery is evaluated:
• by analysis of a Certified Reference Material
conditions where independent test results are (CRM) or
obtained with the same method on identical test • by fortified test portion with analyte and
items in different laboratories with different operator measured the analyte concentration.
using different equipment. Proficiency
testing/collaborative studies between laboratories.

17 18

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Recovery Robustness

Determination of recovery by using blank spike sample: • Intra-laboratory study to check changes due to
• Spike of a known quantity of a substance in a environmental and/or operating conditions.
blank
• sample determination of the recovery. • Carry out at least 6 replicate analyses on a sample
using the analytical method under validation with
• Cfrt: Concentration determined in fortified sample some critical conditions such as:
• Cunf: Concentration determined in unfortified • Small change of pH level
• Extraction time
sample • Temperature of extraction
• Cadd: Concentration of fortification • Change of flow rate
• Change of another volatile solvent
19
20

METHOD VALIDATION GUIDELINES Thank you.

• EURACHEM Guide: The Fitness of Purpose of
Analytical Methods : A Laboratory guide to Method
Validation and Related Topics (EURACHEM 1998)

• IUPAC Technical Report : Harmonized Guidelines For
Single Laboratory Validation of Methods of Analysis :
Pure Applied Chemistry Vol 74 No 5 835-855, 2002

• Nordic Committee on Food Analysis: "Validation of
Chemical Analytical Methods" (NMKL Procedure No.
4, 1996)

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CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

COURSEWORK
WITH SAMPLES

CENTRE OF ENVIRONMENTAL HEALTH
AND SAFETY

FACULTY OF HEALTH SCIENCES

ENV 460
ANALYTICAL TECHNIQUE AND INSTRUMENTATION

ASSIGNMENT

Assignment: Review Paper (60%)

Students will be divided into several groups (4-5 people per group). Each group is required to 
conduct a scoping review using Preferred Reporting Items of Systematic reviews and 
Meta-Analyses (PRISMA) guidelines. http://www.prisma-statement.org/ For this assignment, each 
group needs to write a review paper in 3000 – 4000 words regarding the following scope: 
Sampling, extraction method, and analysis of various pollutants in the environment i.e., PAHs, 
endocrine disrupting chemicals, heavy metals, halogenated hydrocarbon, pesticides, etc. Minimum 
20 research articles are required for these reviews. The paper should have the following 
information: Introduction Methodology Results & Discussion Conclusion References 
 
 

 
 

 
 
 

 
 

 
 
 

 
 

 
 
 

 
 

 
 

 
 
 

 
 

 
 















RESEARCH APPROACHES ON THE CONCENTRATION OF
POLYCYCLIC AROMATIC HYDROCARBONS (PAHS) IN FOOD

AND HEALTH RISK ASSESSMENT FROM 2010-2020:
A SYSTEMATIC REVIEW

Name of author (s): N​ urul Raihana, Nur Afiqah Syazwany & Norhayati

Address; C​ entre of Environmental Health and Safety, Faculty of Health Sciences, Universiti Teknologi MARA

(UiTM), UiTM Kampus Puncak Alam, 42300 Bandar Puncak Alam, Selangor, Malaysia

Abstract: T​ his study aims to review the levels of polycyclic aromatic hydrocarbons (PAHs) in

different types of food products and human health risk assessment due to the consumption of the food.
This review included the extraction method, analyzed method, levels of PAHs, and calculation of
human health risk assessment. The results compiled the concentration of PAHs in various food
products, especially smoked and grilled meat, edible oils, bread, and cereals based product.
Solid-phase extraction was the most commonly used extraction method. Besides, HPLC-FD was used
to analyze PAHs. On the other hand, incremental lifetime cancer risk (ILCR), a margin of exposure
(MOE), chronic daily intake, and dietary exposure are types of health risk assessment identified based
on the reviewed papers.

4.1 INTRODUCTION

Polycyclic aromatic hydrocarbons (PAHs) are defined as organic compounds that
contain only hydrogen and carbon. They also consist of two or more fused aromatic rings
(Conte et al. 2016). The arrangement of PAHs can be in the linear, angular, and clustered
form (Liu et al. 2017). The source of PAHs are usually come from incomplete combustion of
organic matter and also during industrial processes. As they have the properties to remain in
the environment for a long duration, then they are also classified as persistent organic
pollutants (POPs). According to the records of The United States Environmental Protection
Agency (USEPA), there are 16 PAHs listed as primary pollutants including naphthalene,
acenaphthylene, acenaphthene, phenanthrene, fluorene, fluoranthene, anthracene, pyrene,
benz[a]anthracene, benzo[b]fluoranthene benzo[k]fluoranthene, chrysene, benzo[a]pyrene,
benzo[ghi]perylene, dibenz[a,h]anthracene, and indeno[1,2,3-cd]pyrene (Krajian and Odeh
2018).

In recent days, dietary exposures have become a significant source of PAHs
contamination to human. PAHs react as contaminants in conventional food products like
vegetables, dairy products, meat products, cereals, and oils and fats (Shariatifar et al. 2020).

The existence of PAHs in these staple food is because of contamination from the
environment, especially in the soil and water, manufacturing process, and food preparation
steps. Moreover, the methods in food processing, such as grilling, roasting, smoking, and
drying, can form and increase the PAHs level in food (Tongo, Ogbeide, and Ezemonye 2017).

However, the risk of contamination of PAHs has risen through the daily intake. PAHs
could pose serious public health concerns, especially cancers, to humans if the residues
accumulated in food over recommended levels. In food, a benzo[a]pyrene and sum of four
PAHs (benzo[a]pyrene, benzo[a]anthracene, benzo[b]fluoranthene, and chrysene) used as an
indicator for PAHs. The international standard, the European Union (Commission Regulation
No. 2015/1125) has set various standards for different food intended for direct consumption
or use as ingredients in food (Commission Directive (EU) 2015). But, a safe threshold of
PAHs exposure in food could not be defined based on the national standard. According to
(Ishizaki et al. 2010), previous studies have shown an association between dietary exposure to
PAHs and an increased risk of human cancers.

This article outlines the determination of potential contamination of PAHs in food as
states in the literature from 2010 to 2020. It was conducted to determine the concentrations of
PAHs in different foods and to assess the health risk associated with consumption.

4.2 METHODOLOGY

Systematic reviews and meta-analyses are essential tools for summarizing evidence
accurately and reliably. A systemic review is an entire process of selecting, evaluating, and
synthesizing all evidence, while meta-analyses are the statistical approach to combine the data
derived from the systematic review. The outlines of a systematic review of Polycyclic
aromatic hydrocarbons (PAHs) in foods in Malaysia showed in Figure 1 based on preferred
reporting items for systematic reviews and meta-analyses (PRISMA) guideline (Moher et al.,
2019). Searches of published literature for PAHs in food conducted between March 2020 and
April 2020, by using the following databases: ResearchGate; Springerlink: ScienceDirect;
NIH; MOH; Public Access; and PubMed. Searches ran by using the keyword, including
"Polycyclic aromatic Hydrocarbons", "Health risk assessment", "PAHs in foods", and
"Malaysia" where the original papers published in local and international journals from 2010
to 2020 were selected.

A total of 145 articles were retrieved through initial search using databases (n=116),
and Google Scholar (n=29). From the initial search, three articles were duplicates and hence
removed. After removing duplicate citations, the selected full text of published sources
retrieved electronically and final selection of relevant sources to include, performed in a
second review, which ensures that the articles complied with the search inclusion and
exclusion criteria. Finally, a total of 24 articles were included in this meta-analysis and
critically reviewed, as summarized in Figure 1.

Figure 1.​ The outlines of a systemic review of PAHs in foods in Malaysia based on Preferred
Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline (Moher et al.,
2009)

4.3 RESULT AND DISCUSSION

4.3.1 Study characteristics of PAHs

Table 4.1 present the study characteristics of PAHs in various food products. In terms
of study population, most of the studies were performed in China (n=4), and followed by
Malaysia (n=3), Kuwait (n=2), Iran (n=2), Italy (n=1), Indonesia (n=1), Poland (n=1), Latvia
(n=1), Africa (n=1), Vietnam (n=1), Syria (n=1) and Korea (n=1).

Table 4.1:

Studies included in the meta-analysis of concentration of PAHs in food from 2010-2020.

Ref Author Year of Location Operating category
ID published

A1 Farhadian et al. 2010 Selangor, Malaysia Grilled meat

A2 Xia et al. 2010 Taiyuan, China Edible oils
A3 Al-Rashdan et al. 2010 Safat, Kuwait Toasted bread
A4 Alomirah et al. 2011 Grilled and smoked foods
A5 Iranda et al. 2012 Kuwait Chicken satay
A6 Jahurul et al. 2013 Yogyakarta, Indonesia Meat and fish products
2013
Ciecierska & Selangor, Malaysia Bakery chain
A7 2015
Warsaw, Poland Edible vegetable oils
Obiedziński
Shandong, China
A8 Jiang et al.

A9 Eslamizad et al. 2016 Tehran, Iran Bread
A10 Rozentale et al. 2017 Latvia Cereals
2017
Abdul-Gafaru Kumasi, Africa Bread
A11 2018
2018 Hanoi, Vietnam Daily food products
Osumanu Iran Edible vegetable oils
A12 Tran-Lam et al.

A13 Yousefi et al.

A14 Krajian & Odeh 2018 Syria Edible oils
A15 Jiang et al. 2018 Shandong Province, China Grilled and fried meat
A16 Lee et al.
2019 Korea products
Edible oils

4.3.2 Method of extraction

As general precautions to be considered when determining PAHs, it is crucial to
protect the solutions against the light as these compounds are light-sensitive, and they can
decompose by photoirradiation and oxidation. Thus, carefully controlled the light exposure
during the sample pre-treatment. Besides, to diminish possible losses due to the evaporation
of the lower molecular weight compounds, the concentration to dryness should be avoided.

In the method of extraction, there are many ways to extract edible oils. Ultrasonication
is an expert technique to extract the high nutritional quality oil directly from the plant-based
origin. This ultra-sonication provides a shorter extraction time as it is easy and safe to use.
Besides, microwave extraction has the same advantages as the extraction using
ultra-sonication. The solid-phase extraction (SPE) is identified to be one of the popular
techniques used for edible oils. This scenario is because the approach is a convenience for
sample preparation in the chromatographic analysis of edible oils. It requires a low volume of
organic solvent and a small amount of sample. Instead of that, it can be accomplished in a
shorter time compared to other approaches.

For the detection of PAHs in edible oils, both chromatographic analysis, liquid
chromatographic, and gas chromatographic can be practical analytical tools. These tools are
useful for the separation of lipids and PAHs contaminants in edible oils.

Moreover, extraction of PAHs from bread has traditionally relied on a three-stage
methodology, including saponification, liquid-liquid extraction (LLE), and clean-up by
column chromatography or, more recently, solid-phase extraction (SPE). Hence, some studies
used the QuEChERS techniques method developed for the analysis of pesticide to extract the
bread for PAHs monitoring. The study of extraction method included Soxhlet extraction +
column chromatography (Al-Rashdan et al., 2010), solid-phase extraction + GPC (Ciecierska
& Obiedziński, 2013), and QuEChERs method (Eslamizad et al., 2016; Abdul-Gafaru

Osumanu, ​2017; Tran-Lam et al., 2018), Sonication + GPC (Rozentale et al., 2017). The
study conducted by Eslamizad et al. (2016), Abdul-Gafaru Osumanu ​(2017), Tran-Lam et al.

(2018) u​ sed QuEChERS techniques; where acetonitrile and deionized water were used as

solvents for extraction.
For the detection of PAHs in bread, gas chromatography and liquid chromatography can

be used to analyses the concentration of PAHs. The gas chromatography of two sensitive
GC-based techniques, GC-MS/MS and GC-HRMS, can be used as it has appropriate
sensitivity and selectivity for PAHs analysis in food products. Furthermore, solid-phase
extraction was mostly used to extract grilled meat and chicken sample, while HPLC-FD
analyzed the detection of PAHs.

​Table 4.2:
​Type of extraction and detection method.

Ref ID Extraction method Detection method
HPLC-FD
A1 Solid-phase extraction + column chromatography GC-MS
GS-MS
A2 Microwave extraction GC-MS
GC
A3 Soxhlet extraction + column chromatography HPLC-FD

A4 Solid-phase extraction + column chromatography HPLC-FLD, GC/MS
HPLC-FD
A5 Solid-phase extraction GS-MS

A6 Solid-phase extraction + column chromatography GS-MS, GS-HRMS
HPLC-FLD
A7 Solid-phase extraction + GPC GS-MS
HPLC-FD
A8 Solid-phase extraction + Liquid-liquid extraction HPLC-UVD
HPLC-FD
A9 QuEChERs method GC-MS

A10 Sonication + GPC

A11 QuEChERs method

A12 QuEChERs method

A13 Ultrasonication

A14 Solid-phase extraction + DACC column

A15 Solid-phase extraction + column chromatography

A16 Solid-phase extraction

4.3.3 The concentration of PAHs in food

The food products that have been studied to detect the concentration of PAHs were
listed in an operating category that consists of meat, chicken, fish, edible oils, bread, milk,
and cereals. The reviewed paper was published from the year 2010 to 2020. Most of the
studies were based on meat, chicken and fish products (Farhadian et al., 2010; Alomirah et al.,
2011; Iranda et al., 2012; Farhadian et al., 2012; Jahurul et al., 2013; Jiang et al., 2018).
Edible oils have been studied by Xia et al., 2010; Jiang et al., 2015; Yousefi et al., 2018;
Krajian & Odeh, 2019 and Lee at al., 2019. Besides, the concentration of PAHs in bread has
been conducted by Al-Rashdan et al., (2010), Eslamizad et al., (2016), and Abdul-Gafaru
Osumanu, (2017). Other food products such as bakery chain, cereals, and daily food products
have been carried out by Ciecierska & Obiedziński (2013), Rozentale et al. (2017), and
Tran-Lam et al., (2018) respectively.

Table 4.3:

The concentration of PAHs in food products.

Ref ID Types of PAHs Samples Concentration
Ayam bakar 28.9-51.1 ng/g
A1 Fln, B(b)P, B(a)P Ikan bakar 9.36-15.0 ng/g
Beef satay 81.0-132 ng/g
A2 B(a)P Chicken satay 14.3-37.6 ng/g
Beef kebab 9.36-15.0 ng/g
A3 16 PAHs Chicken kebab 8.23-20.4 ng/g
Grilled chicken 4.63-12.4 ng/g
Tandoori (chicken) 4.65-9.66 ng/g
Oven grilled chicken 3.51-7.14 ng/g

Fruits 0.33 ng/g
Vegetables 0.14 ng/g
0.28 ng/g
Rice 0.63 ng/g
Wheat 5.71 ng/g
Fish 1.69 ng/g
Pork 0.64 ng/g
Chicken 2.29 ng/g
Beef and mutton 0.26 ng/g
Eggs 0.18 ng/g
Milk 0.69 ng/g
Oils FA 1.19-2.19 µg/kg
White Wheat flour

PY 0.71-1.66 µg/kg

NA 4.33 µg/kg

FL 5.04 µg/kg

PHE 31.4 µg/kg

B(a)P 2.83-16.54

µg/kg

Brown Wheat flour FA 1.19-2.19 µg/kg

PY 0.71-1.66 µg/kg

NA 12.9 µg/kg

FL 8.14 µg/kg

PHE 32.0 µg/kg

B(a)P 11.13 µg/kg

Sandwich bread baked with white FA 1.19-2.19 µg/kg

flour PY 0.71-1.66 µg/kg

B(a)P 2.83-9.04

µg/kg

A4 16 PAHs Meat mandi 24.7-113 µg/kg

Meat kebab 40.8-532 µg/kg

Meat tikka 302- 1292 µg/kg

Meat shawarma 11.3-40.1 µg/kg

Meat burger 15.8-219 µg/kg

Smoked meat 82.9-101 µg/kg

Smoked fish 255-263 µg/kg

Meat arayes 42.3-73.9 µg/kg

Chicken mandi 141-202 µg/kg

Shish tauk 53.4-665 µg/kg

Whole grilled chicken 48.2-342 µg/kg

Chicken shawerma 9.29-36.5 µg/kg

Chicken burger 6.41-23.8 µg/kg

Pita bread plain 7.50-32.1 µg/kg

Pita bread with fat drippings 42.2-554 µg/kg

Pita bread wrapped in shawerma 23.5-52.3 µg/kg

Burger bun 25.0-162 µg/kg

Grilled vegetables 42.9-229 µg/kg

A5 BaP Chicken satay 2.5-393 ppb

A6 Fln, B(b)F, B(a)P Chicken satay 42.31 ng/g

Chicken barbecue 17.89 ng/g

Chicken percik 13.05 ng/g

Chicken tandoori 11.03 ng/g

Chicken honey 8.92 ng/g

Chicken black pepper 6.24 ng/g

A7 19 PAHs Chicken cooked with chili nd
Chicken rendang nd
Chicken golek nd
1.02 ng/g
Chicken in milk gravy nd
Chicken soup 1.28 ng/g
nd
Fried chicken (stir fried) nd
Nugget 8.45 ng/g
nd
Chicken paprik nd
Sausage nd
nd
Chicken burger 10.69 ng/g
Chicken kurma 2.39 ng/g
Chicken curry 9.02 ng/g
nd
Steam fish nd
Charcoal-grilled fish nd
nd
Fried fish nd
Fish three flavor nd
nd
Salted fish nd
Fish in coconut milk nd
nd
Fish curry nd
Fish soup 66.28 ng/g
Fish in tamarind nd
Fish fried in chili 8.98
Fish in taucu sauce nd
Fish in soya sauce nd
Keropok Lekor nd
Beef curry 30.76 ng/g
Beef soup nd
Bees satay 1.07-3.65 µg/kg
Beef kurma 1.07-3.65 µg/kg
Beef paprika 1.07-3.65 µg/kg
Beef burger 1.59-13.6 µg/kg
Beef rendang
Beef in coconut milk gravy
Mutton satay
Mutton curry

Grain
Flour
Bran
Bread

A8 Nap, Ace, Fle, Phe, Ant, Flu, Pyr, Blend oil 3.02-88.55 µg/kg
BaA, Chr, BbF, BaP, BkF, DahA, Corn oil 3.71-185.72 µg/kg
IcdP, BghiP Peanut oil 11.45-151.38 µg/kg

Soybean oil 9.68-331.95 µg/kg

A9 BaP Semi-industrial Sangak bread in LOQ-2.46 ng/g

Shiraz

Traditional Sangak bread in Shiraz LOQ-7.73 ng/g

Traditional Sangak bread in Tehran LOQ-3.19 ng/g

Industrial bread in Tehran LOQ-0.25 ng/g

All bread in Tehran and Shiraz LOQ-7.73 ng/g

A10 BaA, Chr, BbF, BaP Cereals 0.4-0.71 µg/kg

Rye bread 0.23-0.45 µg/kg

Wheat bread 0.21-1.29 µg/kg

Wheat-rye bread 0.05-0.47 µg/kg

A11 Nap, Ace, Ant, Flt, Pyr, BaP, BbF, Butter bread 18.47-2347.34 µg/kg

DBA Sugar bread 1.62-269.65 µg/kg

Tea bread 4.02-76.40 µg/kg

A11 18 PAHs Instant noodles 9.3-9.6 µg/kg

Cakes 0.22-2.48 µg/kg

Dried vegetables 0.91-4.83 µg/kg

Teas 5.14-23.32 µg/kg

Coffees 4.82-24.35 µg/kg

Grilled meats 1.43-25.2 µg/kg

A13 4 PAHs Frying oil 4.70-30.96 µg/kg

A14 4 PAHs Blended oil 3.32-24.35 µg/kg
A15 Nap, Ace, Fle, Phe, Ant, Flu, Pyr, Sunflower oil 5.70-12.78 µg/kg
5.85-27.38 µg/kg
BaA, Chr, BbF, BkF, BaP, DahA, Corn oil 17.65-69.60 µg/kg
BghiP, IcdP Canola oil 0.33-24.3 µg/kg
Edible oils
A16 80.0 µg/kg
4 PAHs Grilled meat products
50.1 µg/kg
Fried meat products
nd-7.46 ng/g
Sesame oil
nd-12.91 ng/g
Perilla oil

Red pepper seeds oil nd-6.26 ng/g

Olive oil 0.42-4.07 ng/g

Red pepper seasoning oil 0.12-6.88 ng/g

Several studies have been conducted to investigate levels of PAHs in grilled or
smoked meat and fish products, as shown in Table 2. The formation of PAHs in food highly
influenced by the cooking process over high temperatures, including grilling and smoking.
Analysis of toxic PAHs (fluoranthene, benzo(b)fluoranthene and benzo(a)pyrene) in grilled
meat by Farhadian et al. (2010) found that beef satay contains the highest level of sum PAHs
and oven grilled chicken contains the lowest with 132 ng/g and 3.51 ng/g respectively.
Alomirah et al. (2011), in their study, stated that meat tikka contained the highest
concentrations of total PAHs, ranging from 302 to 1292 µg/kg. In another study, Irnanda et al.
(2012) mentioned the level of benzo(a)pyrene in chicken satay in Yogyakarta was ranging
from 2.5 to 393 ppb. Furthermore, Farhadian et al. (2012) reported that the highest total
PAHs concentration was found in beef satay (66.28 ng/g), and the lowest was found in
chicken barbecue (17.89 ng/g). Besides, the levels of fifteen PAHs in grilled meat products
were 80.0 µg/kg (Jiang et al., 2018).

Based on the European Union (Commission Regulation No. 2015/1125), the
maximum level for benzo[a]pyrene has been set to 2 µg/kg, and 10 µg/kg for a sum of the
four individual PAHs in edible oils. Xia et al. (2010) reported the content of BaP​eq in edible
oils in Taiyuan, China, is 0.69 ng/g, which showed in compliance with the EU regulation as it
was lower than 2 ng/g. However, the BaPe​ q concentration in other food such as rice, fruits, and
vegetables was not stated in EU regulation. Jiang et al. (2015) measured the soybean oil
contained the highest concentration of PAH1​ 5​, which is 331.95 µg/kg. Rapid urbanization and
economic accelerated development in Shandong, China, located near the soybean oil
production, causing severe pollution of PAHs in this oil. Yousefi et al. (2018) also determined
the concentration of PAHs in edible oils variations surpassed 10 µg/kg, were regarded as
unacceptable according to the EU regulation. Krajian and Odeh (2018) found that the
concentration of PAH​4 in edible oils in Syria did not fulfill the EU food law requirement. Lee,
Suh, and Yoon (2019) also evaluated the concentration of PAH4​ in perilla oil is more than 10
ng/g and not complied with the EU Commission standard.

In Iran, the results reveal that BaP concentrations in traditional bread samples are
higher than the maximum levels set for processed cereal-based foods and baby foods. BaP
content in industrial bread samples was under LOQ. In a study conducted by Al-Rashdan et
al. (2010), the rest of the samples, BaP are varied from 2.83 to 16.54 μg/kg. BaA, CHR,
B(b)FA, B(k)FA, IP, DB(a,h)A, and B(ghi)P concentrations were found to be less than 10.0

μg/kg. However, BaP is not detected in original white and brown wheat flour. Tran-Lam
(2018), in many investigated samples of BaP in instant noodles and grilled meat samples,
shows exceeded level from the maximum limits tolerated by the European Commission (35
µg/kg and 5 µg/kg, respectively). Abdul-Gafaru Osumanu (2017) stated that the concentration
of PAHs in bread samples is expected not to cause a health risk to human consumers in
Africa.

4.3.4 Health risk assessment

Only a few studies focused on health risk assessment associated with the food products
consumed by the population of the study area, as shown in Tables 4.4, 4.5, and 4.6. Incremental
lifetime cancer risk (ILCR) has been calculated by Xia et al. (2010), Jiang et al. (2015),

Yousefi et al. (2018), and Jiang et al., (2018). Abdul-Gafaru Osumanu (2017) ​have calculated

chronic daily exposure (CDI) in his studies. The margin of exposure (MOE) has been
calculated by Yousefi et al., (2018), Krajian & Odeh, (2018) and Lee at al., (2019).

Table 4.5:

Health risk assessment based on Incremental Lifetime Cancer Risk (ILCR).

Ref Incremental Life Time Cancer Risk (ILCR)
ID Children
Adolescents Adults Seniors

Male : 1.19 x 10-​ 5 Male : 7.91 x 10​-6 Male : 4.04 x 10​-5 Male : 8.03 x 10​-6
A2
Female : 7.08 x 10-​ 6 Female : 3.87 x 10​-5 Female : 7.97 x 10-​ 6
Female : 1.16 x 10-​ 5 3.29 x 10​-6 1.08 x 10-​ 5 2.95 x 10-​ 6
- 5.20 x 10-​ 6 -
A8 7.76 x 10​-6 1.14 x 10-​ 6 3.75 x 10-​ 6 1.02 10​-6
A13 3.17 x 10-​ 5
A15 2.69 x 10-​ 6

Incremental lifetime cancer risk (ILCR) is one of the approaches to estimate health risk
associated with the ingestion of carcinogenic toxicants, including PAHs. US EPA stated that the
acceptable level for cancer risk was in the range of 10​-6 ​(permissible level) to 10​-4 (serious level)
(Sultana et al., 2017). Jiang et al. (2018) reported that the calculated ILCR values due to dietary
exposure of grilled and fried meats were 2.69 x 10-​ 6 for children, 1.14 x 10-​ 6 for adolescents. 3.75 x
10-​ 6 for adults and 1.02 x 10-​ 6 for seniors in a 76-year life span. The ILCR value for all groups was
not exceeded the 10​-4 ​limit, indicating the potential cancer risk was slight.

Table 4.5:

Health risk assessment based on chronic daily intake and dietary intake.

Ref ID Samples Chronic Daily Intake Dietary Intake
(CDI)
Tehran 170.6 ng/day
Semi-industrial Sangak bread in Shiraz Shiraz 168.7 ng/day

Traditional Sangak bread in Shiraz

A9 Traditional Sangak bread in Tehran

Industrial bread in Tehran

All bread in Tehran and Shiraz

Butter bread 2.52E-​ 10 a​ t 5%,

A11 Sugar bread 1.87E-​ 9​ at 50%,

Tea bread 2.06E-​ 7​ at 95%

Based on animal carcinogenicity data, an acceptable daily intake of BaP has been
figured out as the quantity that would be associated with a 1/10​6 increase in the risk of cancer
for an adult of 70 kg. This risk increase level corresponds to a dietary intake level of around
0.05 µg/day (Eslamizad et al., 2016). The average dietary exposure in Shiraz and Tehran only
through bread samples was higher than mean dietary consumers across European countries
(0.95 ng/kg b.w. per day) through cereals and cereal products. However, BaP concentrations in
the traditional and semi-industrial samples are less than maximum levels regulated in processed
cereal-based foods and baby foods (Eslamizad et al., 2016). However, a study presented by

Abdul-Gafaru Osumanu ​(2017) in Africa stated that the chronic daily intake for butter bread,

sugar bread, and tea bread is 2.52E-​ 10 ​at 5%, 1.87E-​ 9 a​ t 50%, and 2.06E-​ 7 ​at 95%. The 95%

percentile is lower than the 10​-6,​ in which the cancer risk is in an acceptable range. The risk
might be acceptable if a section rather than the entire population of getting cancer at the higher
end of the range (10​-4 rather than 10​-6​) but not the probability of dying of cancer and
corresponding chronic daily intake (CDI) over an assumed 70-year human lifetime.

Table 4.6:

Health risk assessment based on the Margin of Exposure.

Ref ID Types of oil The Margin of Exposure (MOE)
217 000
Frying oil 233 000
410 000
A13 Blended oil 405 000
-
Sunflower oil 14 900
1 847 826
Corn oil 1 317 829
1 227 437
Canola oil 985 507
1 531 532
A14 Edible oils

Sesame oil

Perilla oil

A16 Red pepper seeds oil

Olive oil

Red pepper seasoning oil

The MOE was used as a new approach to assessing the risk for genotoxic and
carcinogenic PAHs. In this situation, if the MOE values are close to or less than 10,000, it
indicates a potential concern for consumer health and possible action for risk management to be
taken. In contrast, the edible oils with a low concern for consumer health showed the results of

MOE values are higher than 10,000. For edible oil from the Syrian market, the MOE value was
14,900 that indicated this type of oil was safe for consumer health. Both edible oils in Korea
and Iran also showed the values of MOE was more than 10,000 and had a low concern to the
health of the consumer.