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Detection Methods and Advancement in Analysis of Food and Beverages: A

Short Review on Adulteration and Halal Authentication

Chapter · January 2018

DOI: 10.1007/978-981-10-7257-4_36

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Detection Methods and Advancement
in Analysis of Food and Beverages:
A Short Review on Adulteration and Halal
Authentication
Norsuhada Abdul Karim and Ida Idayu Muhamad

1 Introduction
Nowadays, increased consumer awareness of Islam's religious obligations has
created a greater demand for halal food and other consumer goods. The global Halal
market value for trade in Halal foods is estimated at US$547 billion a year. This
large market has created interest from food producing countries worldwide. The total
number of Muslims in Asia in 2014 was about 4.3 billion (32.16 %), while total
Muslim in Australia is 23.1 Million (2.25%) (World Halal Food Council, (WHFC)
2015). These large population and market, has created a great opportunity for halal
food business both domestic and international trade. The global halal food industry is
increasing and this demand develops halal standards, traceability systems, and halal
science centres for halal food detection (van der Spiegel et al., 2012).
Halal is an Arabic and Qur'anic term which means ‘permitted, allowed,
authorised, approved, sanctioned, lawful, legal, legitimate or licit’. Guidelines for
halal are given by Allah in the Holly Quran; “Forbidden to you (for food) are: dead
meat, blood, the flesh of swine and that on which hath been invoked the name of
other than Allah… .” (Surah Al-Ma’idah, 5: 3). Halal term is used in relation to food
and other consumer goods, means “allowed be eating and using by Muslims”.
Haram is the opposite of halal while Shubhah or Mashbooh, means doubt or
suspicion. Food and ingredients that considered haram for Muslim consumption can
be classified in four types such as carrion, pig and derivatives, alcohol and

N. A Karim I. I. Muhamad
Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy
Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Malaysia
email: norsuhadakarim@yahoo.com.my

I.I. Muhamad (✉)

IJN-UTM Cardiovascualr Engineering Centre, V01 FBME, Universiti Teknologi Malaysia,
81310 Johor Bahru, Malaysia
email: idaidayu@utm.my

© Springer Nature Singapore Pte Ltd. 2018 397

N. Muhammad Hashim et al. (eds.), Proceeding of the 3rd International Halal
Conference (INHAC 2016), https://doi.org/10.1007/978-981-10-7257-4_36
398 N. A. Karim and I. I. Muhamad

derivatives and blood derivatives or part of human. With technological advances in

the food processing industry, adulteration and fraud have become common due to
monetary benefits.
Generally, the problem of Haram contamination is focused in animal products;
however, the advance of food industrial technology introduces more and more
complexity of raw materials and products into the field. It is then hard to identify or
trace the origin of such materials. The scientific procedure is essential for providing
an answer. In the present world, raw materials of both animal and non-animal origins
have been totally modified and so scientific procedure is required for identifying
their sources, to confirm whether they are Haram or unlawful for Muslim or not. The
basic scientific laboratory services investigating Haram or dubious (Mashbooh)
substances including:-

• Pork and lard – pork is a pig meat, while lard is pig fat in both its rendered and
unrendered forms. Pig and derivatives is Haram as stated in Quran.
• Alcohol – Liquor (khamr) is defined in Islamic Law as any intoxicating drink that
could affect the person’s mental coherence. In Hadith, it means “whatever
intoxicates in large quantity, and then a small quantity of it is also forbidden
• Gelatin – substance derived from skin or collagen of animal or plant. The gelatin
from porcine skin is most used. The technique widely used for quantitatively and
qualitatively analyzing of gelatin is chemical and crystallization reaction.
• Collagen – subcutaneous protein which is widely utilized for beauty purpose and
mainly produced from swine
• Emulsifiers (Monoglyceride and diglyceride) – obtained from digested oils or fats.
It is Haram if it is from unlawful origin. Its fatty acid profile provides sample
informations of origin
• Fats and oils – can be identified for their origin by analyzing their hydrolyzed fatty
acid profiles
• DNA – in order to identify presentation of porcine and/or canine or other fraud
identification or contaminations in food products and ingredients

Therefore, verification and testing of halal products has become one of the major
challenges in the analysis of highly processed foods or beverages. Thus, this paper
review some of the instrumentation used for verification of Halal foods and
beverages products to provides right halal information for consumer uses.

2 Detection Method of Halal Foods/Beverages

Adulteration and Authentication using Different
Instrumental Analysis

Identification of ingredients in processed or composite mixtures and verification

that the components are authentic and from sources acceptable to consumers has
become necessary. Authentication is the process by which a food is verified as
Detection Methods and Advancement in Analysis of Food and… 399

Sample
(Solid/Liquid)

Extraction

Option of Instrumentations

Basic Analysis: FTIR GC-MS DSC HPLC PTR-MS

MALDI-TOF-MS ELISA 1H NMR Biosenso

Advanced
r
Analysis:
Basic+Chemometric Dielectric FTIR-ATR Real-time PCR

Fig. 1: Flow of detection methods of halal food/beverages adulteration

complying with its label description. Authenticity testing and analytical techniques
have been developed, each appropriate and specific to deal with a particular
problem. The most suitable technique for any particular sample is often determined
by the nature of the sample itself, for instance whether it is raw or cooked, whole
foods or comminuted, solid, semi-solid or liquid form (Nakyinsige et al., 2012).
Halal certification has been made mandatory for all foods and beverages to ensure
its quality and reduce false information on labels for consumer goods. There are
many analytical methods currently used to detect halal authentication of foods and
beverages includes; DNA polymerase chain reaction (PCR), Fourier transform-
infrared spectroscopy (FTIR), gas chromatography-mass spectroscopy (GC-MS),
high performance liquid chromatography (HPLC), differential scanning calorimetry
(DSC), Proton transfer reactions-Mass Spectroscopy (PTR-MS), Proton Nuclear
Magnetic Resonance (1H NMR) Spectroscopy, and also sensor such as Electronic
Nose (e-Nose). Figure 1 showed the optional of instrumentations for halal
authentication.

3 Instrumentation and Concept of Applications

3.1 FOURIER TRANSFORM-INFRARED
SPECTROSCOPY (FTIR)

Fourier transform-infrared spectroscopy (FTIR) is a technique which is used to

obtain an infrared spectrum of absorption, emission, photoconductivity or Raman
scattering of a solid, liquid or gas (Griffiths and de Hasseth, 2007). FTIR results can
explain the functional groups of the sample products. Various techniques of FTIR
were applied including near-infrared spectroscopy (14,000 to 4000 cm−1) (NIR),
mid-infrared spectroscopy (4000 to 400 cm−1) (MIR) and far infrared spectroscopy
(400 to 50 cm−1) (Domingo et al., 2013). Of note, MIR and NIR are most commonly
used, while ATR is advanced analysis used for detecting food contaminants since
400 N. A. Karim and I. I. Muhamad

less sample preparation. Infrared spectroscopy has become an attractive alternative

for traditional analytical methods because it requires little sample preparation,
minimal use of hazardous solvents and high sensitivity and specificity which allows
it to serve as a fingerprint technique (Domingo et al., 2013). Table 1 shows a
summary of basic and advanced FTIR analysis in the halal authentication of foods
and beverages products.

Table 1 Basic and advanced FTIR analysis in the halal authentication of foods and beverages
products

Techniques of FTIR Authenticity Types of Food/Beverage Reference

issue
FTIR Lard  Mixed fats Che Man and
(chicken/lamb/cow) Mirghani (2001)
 Cake formulation Syahariza et al.
(2005)
 Cod-liver oil Rohman and Che
Man (2009)
 Biscuit formulation Che Man et al.
(2010)
Porcine  Bovine & Porcine Hashim et al.
(2010)
FTIR-chemometrics Lard  Chocolate products Che Man et al.
(PLS/PCA) (2005)
 Cod-liver oil Rohman and Che
Man (2011a)
 Vegetable Oil Rohman et al.
(2011a)
 Meatball broth Kurniawati et al.
(2014)
Pork  Beef meatball Rohman et al.
(2011b)
 Ham sausages Xu et al. (2012)
Melamine  Milk Domingo et al.
(2013)
FT-NIR Melamine  Infant formula Balabin and
Smirnov (2011)
FT-MIR Melamine  Liquid milk, and milk Balabin and
powder Smirnov (2011)
Lard  Virgin coconut oil Rohman and Che
Man (2011b)
Alcohol  Wine Friedel et al.
(2013)
FTIR-ATR-PLS Lard  Butter Nurrulhidayah et
al. (2015)
FTIR-ATR-PCA Porcine  Bovine, porcine and fish Cebi et al. (2016)
gelatins
Detection Methods and Advancement in Analysis of Food and… 401

3.2 Polymerase Chain Reaction (PCR)

Polymerase chain reaction (PCR) is a technology in molecular biology used to

amplify a single copy or a few copies of a piece of DNA across several orders of
magnitude, generating thousands to millions of copies of a particular DNA sequence
(Viljoen et al., 2005). DNA is relatively stable and is often still present in many
products even after processing of the food products (Grohmann, 2010). PCR, a
specific target sequence is amplified by the designated primers. Furthermore, the
specificity and sensitivity of PCR can be improved by incorporating an internal
hybridization probe (Mohamad et al., 2013). These techniques are proven to be rapid
and specific in detecting species adulteration. The chemistry involved in PCR
depends on the complementarity (matching) of the nucleotide bases in the double-
stranded DNA helix. When a molecule of DNA is sufficiently heated, the hydrogen
bonds holding together the double helix are disrupted and the molecule separates or
denatures into single strands. If the DNA solution is allowed to cool, the
complementary base pairs can reform to restore the original double helix. In order to
use PCR, the exact sequence of nucleotides that flank (lay on either side of) the area
of interest (the target area that needs to be amplified), must be known. This is the
absolute minimum data necessary before a typical PCR reaction can be used (Viljoen
et al., 2005). The DNA can be detected using variables types or techniques using
PCR analysis (Table 2).
Figure 2 shows a significantly high yield genomic and mt-DNA extracted from
meat and fat samples for lard and pork adulteration.

3.3 Gas Chromatography (GC)

Gas chromatography (GC) is also known as vapor phase chromatography (VPC),

or gas–liquid partition chromatography (GLPC) Detectors such as flame ionization
detector (FID), thermal conductivity detector (TCD), electron capture detector
(ECD), and mass spectroscopy (MS), provide a shorter analysis time and lower
elution temperatures of the sample due to higher flow rates and low molecular
weight (Otles, and Ozyurt, 2015). A large number of volatile compounds, such as
esters, alcohols, fatty acids, aldehydes, ketones, hydrocarbons, ethers, sulfur
compounds, alicyclic compounds, aromatic compounds, heterocyclic compounds
and others can be detected by gas chromatography (Riu-Aumatell et al., 2014). The
adulteration of foods and beverages especially in oils, fats and alcohol has been a
problem to Muslim consumers. Gas chromatography coupled to flame ionisation
detection (GC-FID) has been use as a routine method for detection of fatty acids in
oils and fats. This technique detects adulteration by comparing the retention times
and peak areas of the fatty acids which have been derivatized into fatty acid methyl
esters against appropriate standards. A comprehensive list of fatty acids with
various carbon lengths and degrees of saturation has been complied by CODEX
402 N. A. Karim and I. I. Muhamad

Table 2 Type of PCR for DNA detection of authenticity issue

Types of PCR Authenticity Types of Food/Beverage Reference

issue
PCR-RFLP Pork and lard  Meat & fats Aida et al. (2005)
 Meat species Murugaiah
Pork et al. (2009)
Pork and  Pork & Wild Boar Mutalib et al.
wild boar (2012)
Species-specific Pork  Food products (sausages and Che Man et al.
PCR the casings, bread and (2007)
biscuits)
 Meat species Karabasanavar
et al. (2014)
Fraud  Commercial raw ground meat Mousavi et al.
Identification (cattle, sheep, and chicken) (2015)
Species-specific Pork  Poultry meat Soares et al.
duplex PCR (2010)
Multiplex Real- Pork  Chinese blood curds Cheng et al.,
Time PCR Pork and  Meat species (dog, cat, rat, (2014)
meat species pork and monkey) Ali et al. (2014)
 Commercial meat (beef, Ali et al. (2015)
chicken, goat, lamb,
buffalo,duck, pigeon and
quail)
 Expensive fish species
(salmon, cod, tuna, carp, rohu
and tilapia
 Five different halal branded
meatballs
Real-Time PCR Pork  Pork, Chicken, Beef, Mutton Tanabe et al.
& Horseflesh in Foods (2007)
 Commercial meat extracts Farrokhi and
Joozani (2011)
 Beef meatballs Roostita et al.
(2014)
Bovine,  Feedstuffs Pegels et al.
ovine and (2011)
caprine
material
Porcine  Processed food materials Mohamad et al.
(2013)
 Commercial capsule shell Sudjadi et al.
(2016)

2009 (Fang et al., 2013). Structure and composition of fatty acids in fats and oils
could be used as an indicator for determination of the source of lipid. Information on
fatty acids profile is important for health awareness and religious commitment
(Indrasti et al., 2010). In addition, gas chromatography can be used for metabolomic
Detection Methods and Advancement in Analysis of Food and… 403

profiling of alcohols derived from various origin since alcoholic beverage is often
used as an ingredient for flavoring and preservation in the food service industry. As
stated in Islamic Law, lard, pork and alcohol are forbidden for Muslims. Therefore,
the development of GC techniques using various detectors is in great demand for
halal food authentication for identifying the origins of alcohol and source of lipid
present in foods, and to ascertain if it originates from non-halal sources. Table 3

Pork Lard
Fig. 2
Electrophoresis
analysis of DNA
extraction from meat
and fat samples. M-1
kb plus DNA ladder;
1—mutton; 2— beef;
3—chicken meat;
4,5,6 and 7—pork;
8— mutton fat; 9—
beef fat; 10—
chicken fat; 11, 12,
13 and 14— lard.
Adapted from Aida et
al. (2005)

Table 3. Summary of GC analysis using various detectors in the halal authentication of foods
and beverages products

Authenticity
Types of Food/Beverage GC Detector References
issue
Pork Nurjuliana et
 Meat & sausage GCMS-HS
adulteration al. (2011)
Lard GC x Indrasti et al.
 Animal fats
adulteration GC-TOF-MS. (2010)
Fang et al.
 Oils and fats GC-MS
(2013)
Alcohol Wang et al.
 Beverages GC-FID
(2003)
 Red, white and cooking wine,
GC x GC- Farah Azura et
industrial alcohol (ethanol) made from
TOF-MS al. (2009)
sugarcane
Law et al.
 Beverage HS-GC-MS
(2009)
Abdul Hamid
 Fermented Glutinous Rice (Tapai) HS-GC-FID
et al. (2009)
 Home-made drinks from fruits Najiha et al.
GC-FID
stored for 3 days (2010)
Brill and
 Beverages GC-MS Wagner
(2012)
HS-SPME– Riu-Aumatell
 Beer
GC-MS et al. (2014)
404 N. A. Karim and I. I. Muhamad

shows various detectors used in GC for halal authentication of foods and beverages
products.

3.4 High Performance Liquid Chromatography (HPLC)

High-Performance Liquid Chromatography (HPLC) is a modern technique widely
used for both analytical and preparative separation of compounds contained in more
or less complex mixtures of different origin (Fanali et al., 2015). Its application for
the analysis of contaminants or detection of adulteration in foods has attracted much
attention since the technique itself has many advantages. The most important
advantage is the fact that sample components that are not readily volatilized could be
separated easily by HPLC. In addition, it is applicable to highly polar, high
molecular mass, strongly ionic and thermally unstable components in food systems.
The other advantage of HPLC is that derivatization of analyte is not required

Table 4 Summary of HPLC analysis for halal authentication in foods and beverages products

Authenticity Types of Reference

Analysis of results
issue Food/Beverage s
Pork  Raw or fresh products The pork triglyceride (TG) profile was Saeed et
(mutton, beef, pork) and distinctly different from that of beef or al. (1986;
some processed mutton. Pork fat has larger amounts of 1989)
products triglyceride containing saturated fatty
acid at the C-2 position than does the
fat of other meat.
Lard  Processed food Detection of lard in processed foods, Rashood
products (pork, beef, TG-separation and checking genuinity et al.
mutton, chicken and and adulteration was achieved (1995)
turkey fats)
 Deep-fried foods Absence detection of lard Marikkar
contamination in the fried peanut oil et al.
(PTO). The triglyceride profiles of (2003)
peanut oil (PTO) and oil extracted from
the fried peanut (FPTO) look
similar. Only slight differences were
noticed on the existing peaks but this
did not seem to provide any clue for
lard contamination in the fried peanut
products
 Vegetable oils (palm The lard contamination in PKO by a Marikkar
oioil (PO), palm kernel oil visual comparison of TAG profiles of et al.
( PKO), canola oil (CLO) PKO adulterated with different animal (2005)
fats with those of the animal fats.
However, this approach was not useful
for PO and CLO. The combination of
liquid chromatographic data to
multivariate procedures shows
distinguishable grouping of lard-
contaminated samples for all three
oils
Detection Methods and Advancement in Analysis of Food and… 405

as often as in gas-chromatographic analysis (Marikkar et al., 2005). Table 4 shows

some researches of liquid chromatography for halal authentication in foods.

3.5 Differential Scanning Calorimetry (DSC)

Differential scanning calorimetry (DSC) is a thermo-analytical technique to study
the physical behavior during processing and storage of carbohydrates, fats and oils,

Table 5: Summary of detection of lard adulteration using DSC

Authenticity
Types of Food/Beverage References
issue
Lard  Vegetable oils and fats such as virgin olive oil, cocoa Marikkar
adulteration butter, and dietary supplement oils such as cod liver oil, et al.
evening primrose oil, flaxseed oil, borage oil, grape seed (2015)
oil, and pumpkin seed oil
 Refined-bleached deodorized (RBD) palm oil Marikkar
et al.
(2001)
 Genuine lard (GLD), beef tallow (BT), chicken fat (CF) Marikkar
as adulterants in canola oil et al.
(2002a)
 Enzymatically-randomized lard (ERLD) as an adulterant Marikkar
in RBD palm oil et al.
(2002b)
 Fried products (tempeh, chicken and beef) Marikkar
et al.
(2003)
 Animal fat (tallow,beef, lard, chicken) Dahimi et
al. (2014)

Fig. 3 Differential scanning

calorimetry (DSC) cooling
thermograms of (A) genuine
lard (GLD), (B) chemically
randomized lard (CRLD),
(C) beef tallow (BT), (D)
mutton tallow (MT), and (E)
chicken fat (CF). (adapted
from Marikkar et al. 2001)
406 N. A. Karim and I. I. Muhamad

proteins, alcohol/water content, and food packaging. Many scientists have used DSC
to deal with adulteration problems associated with edible fats and oils, and fat-based
products (Table 5) and example of DSC thermograms of animal fats (Fig. 3).

3.6 Proton transfer reactions-Mass Spectroscopy

(PTR-MS)

Proton transfer reaction mass spectrometry (PTR-MS) is a new and emerging

technique for the measurement and monitoring of volatile organic compounds
(VOCs) at low concentrations in gaseous samples in more or less real time (Hewitt et
al., 2003). Van Ruth et al. (2010a) have found the use of PTR-MS combined with
chemometric for measuring volatiles in a variety of animal fats (milk fat, cow fat,
pig fat) and vegetable oils (coconut, palm and palm kernel oils). PTR-MS resulted in
89% correct classifications, has the advantage that it allows very rapid measurements
compared to the other techniques, but requires further studies. Further, Van Ruth et
al. (2010b) focuses on the identity prediction of three by-products of the fat industry
(animal fats, fish oils, recycled cooking oils), which could be used for animal
feeding. Their identities were predicted by their triacylglycerol fingerprints, their
fatty acid fingerprints and their profiles of volatile organic compounds. Partial least
square discriminant analysis allowed samples to be assigned successfully into their
identity classes. Most successful were triacylglycerol and fatty acid fingerprints
(both 96% correct classification). Proton transfer reaction mass spectra of the
volatile compounds predicted the identity of the fats in 92% of the samples correctly.

4. Advancement in Halal Authentication

4.1 Biosensor
4.1.1 Electronic Nose

The electronic nose is a rapid tool for aroma profile analysis in several fields,
including food analysis, medical diagnosis, environmental pollution monitoring,
cosmetics, and the automotive industry (Nurjuliana et al., 2011b). It is preferred to

Table 6: E-nose used in the lard and pork authentication of foods and beverages products

Authenticity
Types of Food/Beverage References
issue
Lard  Refined, bleached, deodorized (RBD) palm olein. Che Man et al.
adulteration (2005)
 Animal fats (Lard and chicken fat, beef fat and Nurjuliana et
mutton fat) al. (2011b)
Pork  Meat & sausages products Nurjuliana et
adulteration al. (2011a)
 Detection Methods and Advancement in Analysis of Food and… 407

routine laboratory analysis because it is rapid, simple and easy-to-handle. Table 6

represents several halal authentications in foods and beverages using e-nose.

4.1.2 Nanobioprobe
Hybrid biomaterials composed of functionalized nanoparticles, covalently or
noncovalently linked to biomolecules, such as peptides, proteins, and
polynucleotides, are particularly interesting and promising for their size-dependent
optoelectronic properties and dimensional similarities to biomacromolecules. These
conjugated biomaterials are potential agents for multiplexed bioassays, material
synthesis, ultrasensitive optical detection and imaging, in vivo magnetic resonance
imaging (MRI), long circulating carriers for targeted drug release, and structural
scaffolds for tissue engineering (Ali et al., 2012). Ali et al. (2012) was structurally
and functionally integrated a 27-nucleotide AluI-cut segment of swine mitochondrial
(mt) cytb gene to a 3nm diameter citrate-tannate-coated gold nanocrystal to fabricate
a novel class of species-specific nanobioprobe to determine pork in ready-to-
consume burger formulations. The method is comparatively cheaper than the real-
time PCR and can be applied to analyze highly compromised heterogeneous samples
where PCR methods may not work due to breakdown of longer DNA template into
smaller fragments.

4.2 Proton Nuclear Magnetic Resonance (1H NMR)

Spectroscopy
Proton nuclear magnetic resonance (proton NMR, hydrogen1 NMR, or 1H NMR) is
the application of nuclear magnetic resonance in NMR spectroscopy with respect to
hydrogen1 nuclei within the molecules of a substance, in order to determine the
structure of its molecules (Silverstein et al., 1991). In samples where natural
hydrogen (H) is used, practically all the hydrogen consists of the isotope 1H
(hydrogen1; i.e. having a proton for a nucleus). A full 1H atom is called protium
(Silverstein et al., 1991).Nuclear magnetic resonance (NMR) spectroscopy has been
extensively utilized in the authentication of olive oil because it requires minimal
sample preparation, shorter analysis time, its nondestructive nature and good
reproducibility when compared to chromatographic methods which are coupled to
mass spectrometry (Fang et al., 2013). Fang et al. (2013) was investigated the
authentication of 10 vegetable oils (extra virgin olive oil, olive oil, canola oil, palm
oil, soybean oil, corn oil, sunflower oil, rice bran oil, peanut oil and coconut oil) with
heating fats (adipose tissues of pig, mutton, beef and chicken) using proton nuclear
magnetic resonance (1H NMR) spectroscopy, gas chromatography–mass
spectrometry (GC/MS) fingerprinting and chemometrics. However, NMR data does
not successfully established a good detection in terms of sensitivity and specificity as
comparable to GC/MS data using partial least squares discriminant analysis (PLS-
DA) and orthogonal projections to latent structures discriminant analysis (OPLS-
DA) as classification models. They also reported the partial least squares (PLS)
408 N. A. Karim and I. I. Muhamad

models were successfully established for the detection of as low as 5% of lard and
beef tallow spiked into canola oil (Fang et al., 2013).

4.3 Competitive Indirect Enzyme-Linked Immunosorbent

Assay (ELISA)
The enzyme-linked immunosorbent assay (ELISA) is an immunological
assay commonly used to measure antibodies, antigens, proteins and glycoproteins in
biological samples. Tukiran et al. (2016) has been developed for rapid detection of
porcine gelatin in edible bird's nest (EBN) using competitive indirect enzyme-linked
immunosorbent assay (ELISA). Three ELISAs were developed by using polyclonal
rabbit antibodies against porcine species-specific amino acid sequences of collagen
a2 (I) chain (pAb1 and pAb2) and a1 (I) chain (pAb3). The limit of detection (IC15)
of the three ELISAs was 0.033, 0.082 and 0.052 mg/mL respectively. The median
inhibitory concentration (IC50) of pAb1, pAb2 and pAb3 was 0.265, 0.394 and
0.228 mg/mL respectively, as well as able to recognise porcine and bovine gelatins.
pAb1 showed slight cross-reactivity with cave nest and egg white, while pAb2
exhibited slight cross-reactivity with blood cave nest and egg white. No cross-
reactivity was observed with EBNs and egg white for pAb3. The recoveries of
porcine gelatin spiked EBNs were in the range of 62.8e125.4% with intra- and inter-
day coefficient of variants (CVs) of 2.9e5.4% and 4.7e9.6% respectively when using
pAb3.Its can be concluded that, pAb3 appeared sufficient for EBN authentication.

4.4 Dielectric Properties from Microwave

The use of electromagnetic radiation in the microwave is a very attractive and
powerful method for characterizing food and some of its components in a non-
invasive way. The interaction between the materials with electromagnetic energy in
the microwave range provides information known as dielectric properties of the
material which consists of the dielectric constant and dielectric loss. A
comprehensive overview of dielectric properties at microwave frequencies for a
wide variety of foods has been reported by many researchers. Dielectric properties
have been utilized to characterize food quality such as water content in food, sugar
content in yoghurt, concentration of acetic acid, grape juice and wine quality and
others. This dielectric measurement and technique provides a simple, rapid, reliable
and also non-laborious alternative compared to current lab-based existing methods
such as FTIR and PCR. The use of this dielectric measurement could significantly
reduce the analysis time required for the determination of a material’s halal status
and also assist to more efficient and reliable decision making processes (Abidin et
al., 2014; Ali et al. 2014b).

Abidin et al. (2014) has developed a potential method for detection and
discrimination of alcoholic containing drinks for halal authentication using dielectric
Detection Methods and Advancement in Analysis of Food and… 409

properties. Behaviors of several pure alcohols, alcohol solution in water and also
liquids with alcoholic contents were studied for verification purpose. Dielectric
constant and dielectric loss factor for low concentration of ethanol solutions were
measured over the microwave frequency from 0.5 to 50 GHz. The measurements
were extended to several commercial alcoholic beverages. From their studied
showed that dielectric properties manage to discriminate alcohol content until the
lowest concentration studied of 0.5% in water mixture at frequency range of 10-25
GHz. Beyond this limit, solution is considered as alcoholic drinks.

4.5 Matrix-Assisted Laser Desorption Ionization–Time of

Flight Mass Spectrometry (MALDI-TOF-MS)

MALDI-TOF-MS is now increasingly common technique used for species

identification and protein profiling. This technique is revolutionary, reliable and
cost-effective is simple and faster than conventional phenotypic and molecular
methods for the identification of human pathogens. Flaudrops et al. (2015) has been
investigated the origin of meat (pork, beef, horse, veal and chicken) and gelatin
(pork or beef) using MALDI-TOF-MS methods. From their finding, MALDI-TOF-
MS was able to detect down to 1% of gelatin in spiked candies and detect down to
20% of pork gelatin in beef gelatin. This proved that this method has potential for
the systematic and routine traceability of meat and collagen.

5. Conclusions
Basic instrumentation is quite fundamental, laboring and time-consuming while
advanced technology resulted in non-destructive, rapid, efficient, high performance,
more precise, practical and reliable analysis. Continuous research and advancement
in instrumentation is crucial for halal authenticity issues and its verification in foods
and beverages. The verification of halal foods and beverages are critical steps to
protect and identify cases of mis-description and overall compliance with Islamic
food legislation. Each methods and instruments has its advantages which
demonstrate the specific information for confirmation of halal status from extracted
food/beverages products, which is nondestructive, simple, and fast.

Acknowledgements The authors were grateful and acknowledge Universiti Teknologi

Malaysia (UTM) for providing the funding support R.J130000.7709.4J115 awarded to
Prof.Dr.Ida Idayu Muhamad. The authors also express their gratitude for the training and
PCR consumable supply from Qiagen Sdn Bhd (Malaysia).
410 N. A. Karim and I. I. Muhamad

References

Abdul Hamid, N. H., Hashim, D. M., Mustafa S., Che Man, Y.B. (2009). Identification of
alcoholic compounds in fermented glutinous rice (Tapai). In: Hashim, P., Ismail, A.,
Sazili, A. Q., Zaki, N.N.M., Arshad, S., Mokhtar, N.F.K. (Eds.) in Proceeding of the
3rd IMT-GT International Symposium on Halal Science and Management. 32 – 36.
Abidin, Z. Z., Omar, F.N., Yogarajah, P., Biak, D. R. A., Che Man, Y.B. (2014). Dielectric
Characterization of Liquid Containing Low Alcoholic Content for Potential Halal
Authentication in the 0.5-50 GHz Range. American Journal of Applied Sciences. 11
(7), 1104-1112.
Aida, A.A., Che Man, Y.B., Raha, A.R. and Son, R. (2007). Detection of pig derivatives in
food products for halal authentication by polymerase chain reaction–restriction
fragment length polymorphism. J Sci Food Agric. 87, 569–572.
Aida, A.A., Che Man, Y.B., Wong, C.M.V.L., Raha, A.R., Son, R. (2005). Analysis of raw
meats and fats of pigs using polymerase chain reaction for Halal authentication. Meat
Science. 69, 47–52.
Ali, M. E., Mustafa, S., Hashim, U., Che Man, Y. B., Foo, K. L. (2012). Nanobioprobe for
the Determination of Pork Adulteration in Burger Formulations. Journal of
Nanomaterials. 1-7.
Ali, M. E., Razzak, M. A., Hamid, S. B. A., Rahman, M. M., Amin, M. A., Rashid, N.R.A.,
Asing (2015). Multiplex PCR assay for the detection of five meat species forbidden in
Islamic foods. Food Chemistry. 177, 214–224.
Ali, M. E., Razzak, M. A., Hamid, S.B.A. (2014). Multiplex PCR in species authentication:
Probability and prospects—A Review. Food Anal. Methods 7, 1933–1949.
Balabin, R. M., and Smirnov, S. V. (2011). Melamine detection by mid- and near-infrared
(MIR/NIR) spectroscopy: A quick and sensitive method for dairy products analysis
including liquid milk, infant formula and milk powder. Talanta. 85(1), 562–568.
Brill, S.K. and Wagner, M.S. (2012). Alcohol determination in beverages using polar
capillary gas chromatography-mass spectroscopy and an acetonitrile internal standard.
Concordia College Journal of Analytical Chemistry. 3, 6-12.
Cebi, N., Durak, M. Z., Toker, O.S., Sagdic, O., Arici, M. (2016). An evaluation of Fourier
transforms infrared spectroscopy method for the classification and discrimination of
bovine, porcine and fish gelatins. Food Chemistry. 190, 1109–1115.
Che Man, Y. B., Abidin, Z., and Rohman, A. (2010). Discriminant analysis of selected edible
fats and oils and those in biscuit formulation using FTIR spectroscopy. Food
Analytical Methods. 1–6.
Che Man, Y. B., Gan, H. L., Nor Aini, I., Nazimah, S. A. H., and Tan, C. P. (2005).
Detection of lard adulteration in RBD palm olein using an electronic nose. Food
Chemistry. 90(4), 829–835.
Che Man, Y. B., Syahariza, Z. A., Mirghani, M. E. S., Jinap, S., and Bakar, J. (2005).
Analysis of potential lard adulteration in chocolate and chocolate products using
Fourier transform infrared spectroscopy. Food Chemistry. 90(4), 815–819.
Che Man, Y., Aida, A., Raha, A., and Son, R. (2007). Identification of pork derivatives in.
food products by species-specific polymerase chain reaction (PCR) for halal
verification. Food Control. 18(7), 885–889.
Che Man, Y.B. and Mirghani, M.E.S. (2001). Detection of lard mixed with body fats of
chicken, lamb, and cow by Fourier transform infrared spectroscopy. Journal of the
American Oil Chemists' Society. 78(7), 753-761.
Che Man, Y.B., Syahariza, Z. A., Rohman, A. (2011). Discriminant analysis of selected
edible fats and oils and those in biscuit formulation using FTIR spectroscopy. Food
Analytical Methods. 4, 404–409.
Cheng, X., He, W., Huang, F., Huang, M., Zhou, G. (2014). Multiplex real-time PCR for the
identification and quantification of DNA from duck, pig and chicken in Chinese blood
curds. Food Research International.
Detection Methods and Advancement in Analysis of Food and… 411

Dahimi, O., Rahim, A.A., Abdulkarim, S.M., Hassan, M. S., Hashari, S. B.T. Z., Mashitoh,
A. S., Saadi, S. (2014). Multivariate statistical analysis treatment of DSC thermal
properties for animal fat adulteration. Food Chemistry. 158, 132–138.
Domingo, E., Tirelli, A. A., Nunes, C. A., Guerreiro, M. C., Pinto, S. M. (2013). Melamine
detection in milk using vibrational spectroscopy and chemometrics analysis: A review.
Food Research International. 60,131-139.
Fanali, C., Dugo, P., Borges, J.-H., Mondello, L., Aturki, Z. and Fanali, S. (2015). Chapter
24: High-performance liquid chromatography: an established separation technique in
food chemistry. In Nollet, L.M.L. and Toldra, F. (Ed.) in Handbook of food analysis.
3rd Edition – Volume II. CRC Press. Taylor & Francis Group, U.S.
Fang, G., Goh, J.Y., Tay, M., Lau, H. F., Li, S.F.Y. (2013). Characterization of oils and fats
by 1H NMR and GC/MS fingerprinting: Classification, prediction and detection of
adulteration. Food Chemistry. 138, 1461–1469.
Farah Azura, K., Hashim, D. M., Che Man, Y.B., Awis Qurni, S. (2009). Volatile profiling of
alcoholic food and beverages and industrial alcohol using GC x GC-TOF-MS for halal
authentication. In: Hashim, P., Ismail, A., Sazili, A. Q., Zaki, N.N.M., Arshad, S.,
Mokhtar, N.F.K. (Eds.) in Proceeding of the 3rd IMT-GT International Symposium on
Halal Science and Management. 1 – 6.
Farrokhi, R. and Joozani, R. J. (2011). Identification of pork genome in commercial meat
extracts for Halal authentication by SYBR green I real-time PCR. International
Journal of Food Science and Technology. 46, 951–955.
Flaudrops, C., Armstrong, N., Raoult, D., Chabrie`re, E. (2015). Determination of the animal
origin of meat and gelatin by MALDI-TOF-MS. Journal of Food Composition and
Analysis. 41, 104–112.
Friedel, M., Patz,C.-D., Dietrich, H. (2013). Comparison of different measurement
techniques and variable selection methods for FT-MIR in wine analysis. Food
Chemistry. 141, 4200–4207.
Griffiths, P. and de Hasseth, J.A. (2007). Fourier Transform Infrared Spectrometry (2nd ed.).
Wiley Blackwell. ISBN 0471194042.
Grohmann, L. (2010). Chapter 7 – Detection of Genetically Modified Plants in Seeds, Food
and Feed. In Frank Kempken, Christian Jung (Eds.) in Genetic Modification of Plants:
Agriculture, Horticulture and Forestry. Springer-Verlag Berlin Heidelberg. Pp.118.
Hashim, D.M., Che Man, Y.B., Norakasha, R., Shuhaimi, M., Salmah, Y., Syahariza, Z.A.
(2010). Potential use of Fourier transforms infrared spectroscopy for differentiation of
bovine and porcine gelatins. Food Chemistry. 118, 856–860.
Hewitt, C.N., Hayward, S. and Tani, A. (2003). The application of proton transfer reaction
mass spectrometry (PTRMS) to the monitoring and analysis of volatile organic
compounds in the atmosphere. J Environ Monit. 5(1), 17.
Indrasti, D., Che Man, Y.B., Mustafa, S., Hashim, D.M. (2010). Lard detection based on fatty
acids profile using comprehensive gas chromatography hyphenated with time-of-flight
mass spectrometry. Food Chemistry. 122, 1273–1277.
Karabasanavar, N.S., Singh, S.P., Kumar, D., Shebannavar, S. N. (2014) Detection of pork
adulteration by highly-specific PCR assay of mitochondrial D-loop. Food Chemistry.
145, 530–534.
Kurniawati, E., Rohman, A., Triyana, K. (2014). Analysis of lard in meatball broth using
Fourier transform infrared spectroscopy and chemometrics. Meat Science. 96, 94–98.
Law, S. V., Dzulkifly , M.H., Fatimah, A. B., Nurul Aimi, R. (2009). Ethanol content of
beverages and condiments in Malaysia. In: Hashim, P., Ismail, A., Sazili, A. Q., Zaki,
N.N.M., Arshad, S., Mokhtar, N.F.K. (Eds.) in Proceeding of the 3rd IMT-GT
International Symposium on Halal Science and Management. 92 – 95.
Marikkar, J.M.N. (2015). Chapter 2: DSC as a valuable tool for the evaluation of adulteration
of oils and fats. In: Chiavaro, E. (ed.) Differential scanning calorimetry applications in
fat and oil technology. CRC Press. Taylor & Francis Group. Florida.
Marikkar, J.M.N., Ghazali, H.M., Che Man, Y.B., Lai, O.M. (2002a). The use of cooling and
heating thermograms for monitoring of tallow, lard and chicken fat adulterations in
canola oil. Food Research International. 35, 1007–1014.
412 N. A. Karim and I. I. Muhamad

Marikkar, J.M.N., Ghazali, H.M., Che Man, Y.B., Peiris, T.S.G., Lai, O.M. (2005).
Distinguishing lard from other animal fats in admixtures of some vegetable oils using
liquid chromatographic data coupled with multivariate data analysis. Food Chemistry.
91, 5–14.
Marikkar, J.M.N., Ghazali, H.M., Long, K., and Lai, O.M. (2003). Lard uptake and its
detection in selected food products deep-fried in lard. Food Research International.
36, 1047–1060.
Marikkar, J.M.N., Lai, O.M., Ghazali, H.M., and Che Man, Y.B. (2001). Detection of Lard
and Randomized Lard as Adulterants in Refined-Bleached-Deodorized Palm Oil by
Differential Scanning Calorimetry. Journal of the American Oil Chemists’ Society.
78(11), 1113- 1119.
Marikkar, J.M.N., Lai, O.M., Ghazali, H.M., Che Man, Y.B. (2002b). Compositional and
thermal analysis of RBD palm oil adulterated with lipase-catalyzed interesterified lard.
Food Chemistry. 76, 249–258.
Mohamad, N.A., Sheikha, A.F.E., Mustafa, S., Mokhtar N.F.K. (2013). Comparison of gene
nature used in real-time PCR for porcine identification and quantification: A review.
Food Research International. 50, 330–338.
Mousavi, S.M., Khaniki, G.J., Eskandari, S., Rabiei, M., Samiee, S.M., Mehdizadeh, M.
(2015). Applicability of species-specific polymerase chain reaction for fraud
identification in raw ground meat commercially sold in Iran. Journal of Food
Composition and Analysis. 40, 47–51.
Murugaiah , C., Noor, Z.M., Mastakim, M., Bilung, L M., Selamat, J., Radu, S. (2009). Meat
species identification and Halal authentication analysis using mitochondrial DNA.
Meat Science. 83, 57–61.
Mutalib, S.A., Nazri, W.S.W., Shahimi, S., Yaakob, N., Sani, N. A., Abdullah, A., Babji,
A.S., Ghani, M.A. (2012). Comparison Between Pork and Wild Boar Meat (Sus scrofa
) by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-
RFLP). Sains Malaysiana. 41(2), 199–204.
Najiha, A.A., Tajul, A.Y., Norziah, M.H., Wan Nadiah, W.A. (2010). A preliminary study on
halal limits for ethanol content in food products. Middle-East Journal of Scientific
Research. 6(1), 45-50.
Nakyinsige, K., Che Man, Y. B., Sazili, A.Q. (2012). Halal authenticity issues in meat and
meat products. Meat Science. 91, 207–214.
Nurjuliana, M., Che Man, Y. B., Mat Hashim, D. (2011b). Analysis of Lard’s Aroma by an
Electronic Nose for Rapid Halal Authentication. J. Am. Oil Chem. Soc. 88, 75–82.
Nurjuliana, M., Che Man, Y.B., Hashim, D.M., Mohamed, A.K.S. (2011a). Rapid
identification of pork for halal authentication using the electronic nose and gas
chromatography mass spectrometer with headspace analyser. Meat Science. 88, 638–
644.
Nurrulhidayah, A.F., Che Man, Y.B., Amin, I., Arieff Salleh, R., Farawahidah, M.Y.,
Shuhaimi, M., Khatib, A. (2015). FTIR-ATR Spectroscopy Based Metabolite
Fingerprinting as A Direct Determination of Butter Adulterated With Lard.
International Journal of Food Properties. 18(2), 372-379.
Otles, S. and Ozyurt, V.H. (2015). Chapter 25: Gas Chromatography. In Nollet, L.M.L. and
Toldra, F. (Ed.) in Handbook of Food Analysis. 3rd Edition – Volume I. CRC Press.
Taylor & Francis Group, U.S.
Pegels, N., González, I., Martín, I., Rojas, M., García, T., Martín, R. (2011). Applicability
assessment of a real-time PCR assay for the specific detection of bovine, ovine and
caprine material in feedstuffs. Food Control. 22, 1189-1196.
Rashood, K. A., Shaaban, R. R. A., Moety, E. M. A., & Rauf, A. (1995). Triacylglycerols-
profiling by high performance liquid chromatography: a tool for detection of pork fat
in processed foods. Journal of Liquid Chromatography, 18, 2661.
Detection Methods and Advancement in Analysis of Food and… 413

Riu-Aumatell, M., Miró, P., Serra-Cayuela, A., Buxaderas, S., López-Tamames, E. (2014).
Assessment of the aroma profiles of low-alcohol beers using HS-SPME–GC-MS.
Food Research International. 57, 196–202.
Rohman, A., and Che Man, Y. B. (2009). Analysis of cod-liver oil adulteration using Fourier
transform infrared (FTIR) spectroscopy. Journal of the American Oil Chemists'
Society. 86(12), 1149–1153.
Rohman, A., and Che Man, Y. B. (2011a). Application of Fourier transform infrared (FT-IR)
spectroscopy combined with chemometrics for authentication of cod-liver oil.
Vibrational Spectroscopy. 55(2), 141–145.
Rohman, A., and Che Man, Y. B. (2011b). The use of Fourier transform mid infrared (FT-
MIR) spectroscopy for detection and quantification of adulteration in virgin coconut
oil. Food Chemistry. 129(2), 583–588.
Rohman, A., Che Man, Y.B., Hashim , P., Ismail, A. (2011a). FTIR spectroscopy combined
with chemometrics for analysis of lard adulteration in some vegetable oils. CyTA -
Journal of Food. 9:2, 96-101.
Rohman, A., Sismindari, Erwanto, Y., & Che Man, Y. B. (2011b). Analysis of pork
adulteration in beef meatball using Fourier transform infrared (FTIR) spectroscopy.
Meat Science. 88(1), 91–95.
Roostita, L.B., Lengkey, H.A.W., Suryaningsih, L., Rachmawan, O., Putranto, W.S.,
Wulandari, E., Utama, G. L. (2014). Beef Meatballs Adulteration Tests with Real
Time Quantitative PCR Detection For Halal Authentication - Case Studies Sellers at
Traditional Market and Small Medium Enterprises (SMEs) Merchants in Indonesia.
AgroLife Scientific Journal. 3(2), 66-68.
Saeed, T., Abu-Dagga, F., & Rahman, H. A. (1986). Detection of pork and lard as adulterants
in beef and mutton mixtures. Journal of the Association of Official Analytical
Chemists, 69, 999–1002.
Saeed, T., Ali, S. G., Rahman, H. A. A., & Sawaya, W. N. (1989). Detection of pork and lard
as adulterants in processed meat: liquid chromatographic analysis of derivatized
triglycerides. Journal of the Association of Official Analytical Chemists. 72, 921–925.
Silverstein, R.M., Bassler, G.C., Morrill, T.C. (1991). Spectrometric Identification of Organic
Compounds, 5th Ed., Wiley.
Soares, S.,. Amaral, J. S., Mafra, I., Beatriz, M., Oliveira, P.P. (2010). Quantitative detection
of poultry meat adulteration with pork by a duplex PCR assay. Meat Science. 85, 531–
536.
Sudjadi, Wardani, H.S., Sepminarti, T., Rohman, A. (2015). Analysis of porcine gelatin DNA
in commercial capsule shell using real-time polymerase chain reaction for halal
authentication, International Journal of Food Properties. 1-18.
Syahariza, Z.A., Che Man, Y.B., Selamat , J., Bakar, J. (2005). Detection of lard adulteration
in cake formulation by Fourier transform infrared (FTIR) spectroscopy. Food
Chemistry. 92, 365–371.
Tanabe, S., Hase, M., Tano, T., Sato, M., Fuijimura, T., Akiyama, H. (2007). A real-time
quantitative PCR detection method for pork, chicken, beef, mutton and horseflesh in
foods. Biosci. Biotechnol. Biochem. 71(12), 3131-3135.
Tukiran, N.A., Ismail, A., Mustafa, S., Hamid, M. (2016). Determination of porcine gelatin
in edible bird's nest by competitive indirect ELISA based on anti-peptide polyclonal
antibody. Food Control. 59, 561-566.
Van der Spiegel, M., van der Fels-Klerx, H.J., Sterrenburg, P., van Ruth, S.M., Scholtens-
Toma, I.M.J. and Kok E.J. (2012). Halal assurance in food supply chains: Verification
of halal certificates using audits and laboratory analysis. Trends in Food Science &
Technology 27, 109-119.
Van Ruth S.M., Villegas, B., Akkermans, W., Rozijn, M., van der Kamp H., Koot, A.
(2010a). Prediction of the identity of fats and oils by their fatty acid, triacylglycerol
and volatile compositions using PLS-DA. Food Chemistry. 118, 948–955.
Van Ruth, S.M., Rozijn, M., Koot, A., Garcia, R. P., van der Kamp, H., Codony, R. (2010b).
Authentication of feeding fats: Classification of animal fats, fish oils and recycled
cooking oils. Animal Feed Science and Technology. 155, 65–73.
 414 N. A. Karim and I. I. Muhamad

Viljoen, G. J., Nel, L. H., Crowther, J. R. (2005). Chapter 1- Background. In: Viljoen, G. J.,
Nel, L. H., Crowther, J. R. (Eds.) in Molecular diagnostic PCR handbook. Springer,
Netherlands.
Wang, M.-L., Choong, Y.-M., Su, N.-W., Lee, M.-H. A (2003). Rapid method for
determination of ethanol in alcoholic beverages using capillary gas chromatography.
Journal of Food and Drug Analysis. 11(2), 133-140.
World Halal Food Council, WHFC (2015). International Halal Conference and Semi Annual
General Meeting WHFC, 5-17 May 2015, EastPreston Islamic College, Melbourne.
Xu, L., Cai, C.B., Cui, H.F., Ye, Z.H., Yu, X.P. (2012). Rapid discrimination of pork in
Halal and non-Halal Chinese ham sausages by Fourier transform infrared (FTIR)
spectroscopy and chemometrics. Meat Science. 92, 506–510.

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