Food Technology Ascertaining Supply Chain Integrity

1.

Overview
The dimension of food integrity widened after many food scandals emerged, not only focus-

ing on the scope of nature, substance, and quality and safety of food but also concerning the foodproduction, including “the way it has been sourced and distributed and being honest about thoseareas to consumers”（Elliot, 2014）. Among the incidents, the horsemeat scandal in Europe, thecontamination of melamine in infant formula milk in China, and the mislabeling of fish species in
the United States hit the sensation of food integrity. The main interest of food integrity is in foodsupply chain management. However, it can be categorized into developing quality management,
food safety management systems, and food integrity management systems（Manning, 2017）. For
instance, developing food integrity management systems into the broader food supply chain
aimed firstly to guarantee safety, quality, and authenticity ; secondly to ensure reliable labelling,
and thirdly to ensure effective management of provenance such as halal status（Manning, 2017）.
After all, the main elements of food integrity should be concerned with product, process, people,
and data integrity. The example of each component in food integrity has been tabulated in Figure
1.

Fig. 1

2.

Examples of each element of food integrity（Manning, 2017）

Food product integrity
Food integrity denotes that all people can have access to safe, authentic, and nutritious foodat all times. In this chapter, the focus is given on the role of food science and technology in verifying food product integrity. Food integrity consists of several elements : quality, safety, authenticity, traceability, and adulteration.
Food quality can be measured based on the food characteristics acceptable to the consumer.
The physical characteristics of food include texture, flavor, and appearance of food products suchas size, shape, color, and grade. Chemical characteristics are related to the nutritional content of187

the food product. In addition, the high quality of food that meets the standards set up in legislation and product specifications is also related to ethical and sustainable production concerning theconsumerʼs acceptability.
Food safety can be defined as the manufacturing practice of preparing, handling, producing,and storing food to avoid contamination and maintain the nutritional value of food in supporting ahealthy diet（FAO, 2004 ; Sheikha, 2015）. Any biological, chemical or physical agent that cancause any illness or injury without control is considered a food hazard. Biological hazards thatcan result on the foodborne illness outbreaks are the major concern in the food industry. The
common organisms that can cause infection, intoxication, and even death include Salmonella, E.
coli, and Clostridium botulinum. Chemical agents such as pesticides and heavy metals that cancause hazards in food can be categorized into three groups :（i）natural chemicals ;（ii）chemicals
used in formulation for food products, or（iii）chemicals that are unintentionally or incidentallypresent in the finished product. Physical agents that can cause physical hazards are not limited tobone, glass, wood, faces, etc. Therefore, risk assessments and suitable mitigation measurements
can be carried out based on the research and previous data on food safety hazards.
Generally, the purpose of food fraud is to gain financially by the enhancement of the apparent value of the product or reducing the production cost. It can be classified as adulteration, tampering, product overrun, diversion, theft, and counterfeiting. The adulteration or contamination offoods is considered if the valuable component in the food is removed or replaced with a cheaperor poor quality to concealed with an actual food product or substitutes instead of expensive ingredients（Goyal et al., 2022 ; Roy & Yadav, 2022）. Food products can be easily adulterated since
it has a lengthy, complicated processing and supply chain. Therefore, food authentication is important to ensure the safety and quality of food products and consumer protection. It is also important for both official bodies and industries in charge of labelling and raw materials and finishedproducts to be tested for compliance with the national legislation, international standards, andother guidelines and specifications（Baeten & Dardenne, 2002）.
The high food adulteration incidence was reported in the literature and can be categorized
into six groups which were ʻanimal origin and seafoodʼ for milk and dairy products, meat andmeat products, fish and seafood ; ʻoils and fatsʼ ; ʻbeveragesʼ for fruit juices, coffee and tea, alcoholic
beverages ; ʻspices and sweetenersʼ for spices and extracts, sweeteners including honey ; ʻgrainbased foodʼ for cereals and pulse ; and ʻothersʼ for organic foods and dietary supplements（Honget al., 2017）. The key players, including the food industry, academia, and government, are awareof this serious issue and have implemented efforts to ensure food integrity, including methods andprocedures. A competent, fast, and reliable analysis is needed to overcome the challenges and ensure product quality. The authentication and adulteration of food can be validated by various
technique including physical, chemical/ biochemical, or DNA molecular depending on the type ofadulterant to be detected（Bansal et al., 2017）. This use of analytical methods such as non-destructive radiative evaluation, fingerprinting characterization, novel technology for rapid detection, and authentication were reviewed.

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3.

Physical technique
The authenticity of food products can be examined by analyzing the physical characteristics

of food such as color, texture, solubility, bulk density, morphological traits, etc（Bansal et al.,2017）
. Physical techniques such as microscopic and macroscopic can be used for the physicalcharacterization.
In the early years, Louveaux et al.,（1978）detected honey adulteration with cane sugar andcane sugar products through optical microscopy. Combining microscopy and macroscopy withchemical profiling also identifies and authenticates herbs and medicinal plants（Sheorey & Tiwari,
2011）. The utilization of microscopy and macroscopy helps detect microbial contamination in foodsuch as fungi. Furthermore, the botanical origin can be detected with the advent of electron microscopy. For instance, the origin of honey can be determined by analyzing the surface pattern ofpollen from honey（Jones & Bryant, 2014）.
4.

Chemical/biochemical technique
In general, the chemical/biochemical techniques offer more accurate results, and the foodʼscontaminants can be detected even at lower concentrations. However, these techniques are costly, time-consuming and require well-trained operators to conduct the experimental works. The
chemical/biochemical techniques can be categorized into four main categories :（1）spectroscopic-based,（2）chromatographic-based,（3）immunologic-based, and（4）electrophoretic-based techniques. The focus of this section is given to the spectroscopic-based and chromatographic-based
technique since these two techniques are the most used especially in food adulteration and authenticity.
4. 1

Vibrational spectroscopic techniques
A range of vibrational spectroscopic techniques such as Fourier transform near-infrared

spectroscopy （FT-NIR）, mid-infrared （MIR）, Raman spectroscopy, and hyperspectral imaging（HIS）has been successfully used as sensitive and fast analytical techniques for the determinationof food contamination. Figure 2 shows the typical vibrational spectroscopic techniques used forfood authentication. The main advantages of these techniques are being non-destructive, relatively low analysis cost, being adopted for both qualitative and quantitative analysis, and providing analternative to wet-chemical and time-consuming techniques（Lohumi et al., 2013）.
NIR region is between 780 to 2500 nm and can detect overtone and combination bands forthe chemical bonds between light atoms, such as C-H, O-H, and N-H that have high vibrationalfrequencies（Osborne et al., 1993）. FT-NIR can be used for qualitative and quantitative analysis
with little or no sample preparation（Lohumi et al., 2015）. Continuous development makes thistechnique reliable and efficient, and it has become a standard analytical method for quality control measurements. FT-NIR can be categorized into three groups based on instrumental devices :（i）sequential instruments - the absorbance is sequentially collected in time, and the instrument

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Fig. 2

Schematic diagram of typical vibrational spectroscopic techniques.

is equipped with a monochromator or filters ;（ii）Fourier transform or multiplex instruments several frequencies are detected simultaneously in the form of an interferogram ; and（iii）multichannel instruments - several detectors separately detect the absorbance at several wavelengths（Lohumi et al., 2015）.
Numerous studies on detecting food adulterations have been conducted using this technique.
For instance, identification of animal meat muscles（Cozzolino & Murray, 2004）, detection of beef
hamburger adulteration（Ding & Xu, 2000）, adulteration in crabmeat and species authenticity（Gayo et al., 2006）. Besides that, this technique also successfully detected protein adulteration in
yogurt and melamine contamination in milk powder（Haughey et al., 2013）and soya bean meal
（Xu et al., 2013）. NIR spectroscopy can be used in different spectral modes, including reflectance,transmission, interactance, and transflectance, to estimate the sampleʼs external and internal
properties. NIR reflectance spectra（1100―2498 nm）managed to detect 10% pulp wash in orange
juice and sugar-acid mixtures with 90% accuracy（Downey et al., 2016）. The transmission mode
offers the most straightforward sampling technique for analyzing solid, liquid, and gas samples. It
can be used to detect a sampleʼs external and internal qualities. For example, this mode has beenused to differentiate white wine based on the origin（Daniel Cozzolino et al., 2003）and detectionof fatty acid composition in beef and chicken（Riovanto et al., 2012）. The interactane mode isused for the internal information of a sample when the transmission measurements are challenging to obtain. The transflectane mode is the combination of transmission and reflectance modes.
It is specifically designed to measure the spectra of thin or clear sample.
FT-IR focuses on the MIR region between 4000 to 400 cm ― 1 of the electromagnetic spectrum.
It can produce a chemical profile of the sample through the fundamental vibrational and rotational stretching modes of molecules. FT-IR provides more excellent chemical information of the190

Food Technology Ascertaining Supply Chain Integrity

sample compared to NIR spectroscopy because it measures fundamental vibrations instead of theovertones and combinations band measured in the NIR region（Lohumi et al., 2015）. The MIR region can be divided into two, which known as the functional group region（4000 to 1500 cm ― 1）
and the fingerprint region（1500 to 500 cm ― 1）.
FT-IR also enables the measurement of all types of samples by using different measurement
modes such as attenuated total reflectance（ATR）, diffuse reflectance, high-throughput transmission（HTT）, and transmission cell. ATR-FT-IR has been widely used in the analysis of food authentication because it can analyze both qualitative and quantitative analysis. ATR demonstrated
a better sampling method with higher accuracy than HTT for a sampling technique such as microbial spoilage in milk（Nicolaou & Goodacre, 2008）. Various combinations of FT-IR with sampling techniques have been successfully applied for adulteration detection in honey. FT-IR spectroscopy has also been combined with chemometrics to analyze high-value edible oils such as
olive oil, avocado oil, etc. The adulteration in edible oils can be detected through a distinct classification due to the difference in fatty acid and triglyceride content（Christopoulou et al., 2004）.The
determination of sugar concentration in fruit juice also can be analyzed using FT-IR spectroscopy
at the fingerprint region of MIR because this region has high sensitivity spectral region as compared to NIR. This suggested FT-IR is a versatile and powerful analytical method for food applications. FT-IR and NIR are compliments to each other with advantages and disadvantages foreach technique.
4. 2

Chromatographic techniques
Among fingerprint techniques, chromatography is found to be more informative in finger-

print analysis. The chromatographic method will provide information by translating the peak intensity presenting the area or height and their position（retention time）. This method is convenient and reliable to characterize the food sample particularly for the used in identifying foodauthenticity. This is because, this chromatographic technique promotes vast and remarkable capabilities with high sensitivity, reproducibility, and robustness that enable them to provide thehighest information content of the food products（Esteki et al., 2018）.
Previously, using various types of detectors, the chromatographic technique has been used to
identify chemical adulteration markers such as fatty acids, oligosaccharides, and tocopherols（Fanali et al., 2016）. Most HPLC or GC methods were developed to detect carbohydrates, carotenoids, amino acids, phenolic or other organic compounds（Herrero et al., 2017）. For instance, thechromatographic technique could detect the adulteration in oil and characterize the blendʼs composition of oils by determining triacylglycerols（TAGs）, sterols, and fatty acids compounds present in the oil（Wernig et al., 2018）.
The most widely used in detecting the adulteration or identification of the authenticity offood products are gas chromatography （GC） and high-performance liquid chromatography（HPLC）. GC-FID is a popular chromatography mode due to being cheap, well-established, robust,and rapid. GC method was developed to detect adulteration in olive oil by comparing the fatty
acid composition and chemometric analysis. GC method can be used to characterize and differentiate different vegetable oils by focusing on the fatty acid methyl esters （FAME）, obtained191

through trans-esterification of vegetable oils（Brodnjak-Vončina et al., 2005）. For instance, theidentification of extra virgin olive oil, which is an adulterer with corn, peanut, rapeseed, and sunflower oils, was successfully demonstrated by Yang et al.（2013）using the GC-MS method. GC
with surface acoustic wave detector（GC-SAW system）combined with chemometrics was also
used to identify the presence of lard in virgin coconut oil（Mansor et al., 2011）. Besides that, theadulteration of extra virgin olive oil with 5% soybean oil has also been identified using the GC
method by analyzing the fatty acids composition as an indicator of purity and the linoleic acidcontent as a parameter for the detection（Jabeur et al., 2014）.The adulteration of almond powder
with the apricot kernel was also detected using GC fatty acid fingerprinting combined with chemometric methods（Esteki et al., 2017）. Nevertheless, frequently, GC needed derivatization to become the main limitation of GC.
This limitation can be overcome by using HPLC, which does not need any derivatization andcan analyze both polar and non-polar compounds. HPLC has successfully demonstrated the adulteration of ovine, caprine cheese, and milk with bovine milk at lower levels（2 % v/v）by analyzing the whey protein ㌼-lactoglobulin（Chen et al., 2004）. Veloso et al.（2002）also detected bovine,
ovine, and caprine milk using reversed-phased HPLC. HPLC method managed to classify winesfrom different denominated areas of origin in the Canary Islands, Spain, by analyzing 15 polyphenol concentrations combined with principal component analysis（PCA）and linear discriminant
analysis（LDA）（Rodríguez-Delgado et al., 2002）. The combination of PCA and LDA, as well as
HPLC equipped with a UV detector, was also used to classify some Australian wines based on
geographical groups with accuracy in the 75 to 100 % range（Bellomarino et al., 2009）. In addition, reversed-phased HPLC combined with PCA and LDA was used to discriminate different varieties of green and roasted coffee beans by analyzing the triglyceride and tocopherol composition（González et al., 2001）. The current trend shifting from HPLC to UHPLC due to the particle sizeof the stationary phase, sample size, and mobile phase have been reduced. Hence, it becomes
more efficient, with high selectivity, throughput, speed, and resolution.
4. 3

Electric nose
Most detection methods for food authentication are time-consuming and costly and require

professional operators to conduct the analysis. As the adulteration scandals become intense recently, a real-time rapid and reliable detection method is needed to tackle this issue. This rapid
detection technique can be used for commercialized processed foods in the market. One of thetechnologies invented is the electric nose（e-nose）, which mimics the human nose as an artificial
olfactory system or machine olfaction for automated simulation of the sense of smell（Roy & Yadav, 2022）.
E-nose is classified as a non-destructive method widely used in many applications, including
environment monitoring, quality control assessment in agricultural goods, fraudulent adulteration,and spoilages. This method can differentiate the quality of food products by detecting the specificcomponents of an odor and analyzing its chemical makeup to identify it. This detection is preferred to routine laboratory analysis because of easy handling of the system, cost effective, andshort time analysis（Roy & Yadav, 2022）.

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Fig. 3 The typical block diagram of an e-nose consists of : 1 synthetic air cylinder, 2 blower, 3 air
inlet valve, 4 closed sample container, 5 sample, 6 volatiles, 7 sampling valve, 8 sensory arraychamber, 9 data processing and pattern recognition, 10 purge valve, 11 purge outlet.（Source : Roy& Yadav, 2022）.

Many research related to food quality using this e-nose has been reported previously. The
idea of an e-nose system for aroma discrimination based on chemical sensors has been demonstrated by Persaud & Dodd（1982）. In general, e-nose consists of four main units :（i）an electronic multi-sensor array system for chemical detection ;（ii）an information processing unit ;（iii）adigital pattern recognition algorithm system that is capable of recognizing odors ; and（iv）a reference library database. Figure 3 illustrates the typical block diagram of an e-nose.
The mechanism of e-nose is based on the identification of volatile organic compounds generated from the food samples in the translation to voltage differences which are detected by limited
selective sensors. Commonly used sensors are surface acoustic waves（SAW）, and conducting
polymer and metal oxide semiconductors（MOS）. MOS sensor has been widely used because of
their high chemical stability, low response to moisture, long life, and reasonable price. The synthetic air（99.9 % pure）blows the generated volatile organic compounds from the sample to thesensor array chamber. The air inlet and sampling valves remain open during the measurement,
whilst the purging valve will be closed to introduce the generated volatile organic compounds tothe sensor array chamber. The received analogue signal by the sensor array which consists of
selective and sensitive gas sensors coupled with a data acquisition card will then convert to digital signals which can be read on the monitor（Roy & Yadav, 2022）. In principle, food quality can
be classified by differentiating the complex odors from food samples generated as voltage response value（output）.
An e-nose has been widely used in detecting various food adulteration including honey, milk
and dairy products, fruits, meat, fish, spice, edible oils, alcoholic and non-alcoholic drinks, tea andcoffee, etc. The examples include the adulteration of edible plant oils with cheaper sources of oils,
fats, and sterols, which have nearly similar fatty acid profiles. Sometimes, adulteration of edible193

oils might cause serious health problems such as Spanish olive oil syndrome（Clemente & Cahoon, 2009）. However, this e-nose technology is trustworthy and highly sensitive to ascertain thepurity and authenticity of the edible oil. The authenticity of edible oils can be determined throughvarious e-nose systems that have been developed. Hai & Wang（2006）detected the adulterationof Camellia seed oil with maize oil using an e-nose device with 10 MOS sensors. The adulterationof argan oil with sunflower oil using 5 MOS sensor-based e-nose（Bougrini et al., 2014）. Besides
that, Yu et al.（2007）also demonstrated the adulteration of skim milk with water and reconstituted milk powder using an e-nose based on MOS sensors. The gas chromatography-based e-nose
was used to inspect the adulteration of honey with corn and rice syrup（Gan et al., 2016）. For
commodities like meat, aroma is the main indicator to be used in e-nose technology. This is because meatʼs lowered shelf-life stability made the aroma more vulnerable. An e-nose has beensuccessfully used to detect the adulteration of various meats. Tian et al.,（2019）reported theminced mutton was adulterated with pork using a PEN 2 e-nose system containing ten selective
MOS gas sensors based on the odor fingerprint. Wang et al.（2019）also successfully detected theadulteration of mutton with duck meat using SPME for volatile compound extraction and PEN 3
e-nose system comprised of 10 MOS sensors. Another common application of e-nose is the detection of adulteration in spice. Banach et al.（2012）investigated the adulteration of ʼsaveloyʼ andʼsausageʼ spices with an admixture of 20% concentration level of adulteration with curry leaves
and garlic spice powder using 38 MOS sensor-based commercial type e-nose（KAMINA, Yson
GmbH）
.
5.

DNA molecular-based technique
DNA is a unique organism, hence, DNA-based techniques can be an effective tool in foodadulteration detection. Unlike physical, chemical and biochemical techniques, this DNA-based
technique can provide an accurate result quantitatively and qualitatively. Therefore, the developed techniques such as multiplex-PCR, quantitative PCR（qPCR）, DNA sequencing, restriction
fragment length polymorphism （RFLP）, randomly amplified polymorphic DNA （RAPD）, andsimple sequence repeats（SSR）or microsatellites methods are the potent technology which arerequired by countries and international trade organizations to agree especially on the standard.
For instance, the DNA-based molecular technique can be used to detect adulteration, particularly
for food with nearly similar physical characteristics. New tools, including high-resolution melting（HRM）PCR, droplet digital PCR（ddPCR）, isothermal amplification（e.g., loop-mediated isothermal amplification, recombinase-polymerase amplification, strand-displacement amplification, helicase-dependent amplification, and rolling-circle amplification）, and next-generation sequencing（NGS）have emerged with a better performance in terms of sensitivity, specificity, speed, andmultiplex.
5. 1

PCR and qPCR
Detection of adulteration and identification of animal and plant species can be carried out us-

ing high sensitivity, specificity, and relatively fast PCR techniques. The simultaneous identifica-

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Food Technology Ascertaining Supply Chain Integritytion of several species can be obtained through multiplex PCR assays using specific primers.
Thus, it can detect and distinguish the species in the food products（Böhme et al., 2019）. This
technique allows the detection of a target in a complex matrix at a very low concentration
through the exponential amplification of a target DNA molecule using a thermal cycler that generates up to millions of copies（Bohme 2019）. Multiplex PCR assays targeting the sensitive qualitative region of mitochondrial DNA have been developed to simultaneously detect different animal species in meat because this region has a high cell copy number. For example, theadulteration of meatballs with meat from dog, rat, rabbit, and squirrel were identified down to0.1%（Ahamad et al., 2017 ; Rahman et al., 2014）. Nevertheless, the limitation of the PCR technique is not reliable for quantification because the copy numbers per cell change between species,
individuals, and even tissues within the same individual.
The quantification measurement in food adulteration is very important to justify whether it
is intentional or unintentional mixing. In this sense, qPCR is a reliable and convenient method toquantify adulterated food products. The principle of qPCR is amplification of nucleic acids, which
can be monitored in real-time, and quantification by measuring the fluorescence coming from therelease of double-strand-DNA-binding dye, which is measured in each PCR cycle（Böhme et al.,2019）
. The performance of qPCR offers high sensitivity, multiplexing, high speed, and relatively
low cost compared to the conventional method. Furthermore, this technique does not require thepost-PCR processing steps like in the conventional method because it is based on real-time monitoring of the increasing number of target DNA molecules. This qPCR technique has successfully
detected the presence of forbidden meat species like pork in raw and processed halal food products（Karabasanavar et al., 2014 ; Sakalar et al., 2015）. Besides that, it has been used in the seafood industry to detect fraud or mislabeling in tuna products（Liu et al., 2016）. However, the effects of tissue composition and matrix components on the PCR efficiency and precision are themain limitations on the absolute or relative quantification.
5. 2

DNAFoil
DNAFoil is a novel technology based on DNA-approach can be used for food adulteration de-tection. It is portable, completely self-administered, and can be performed on-site without expensive equipment such as PCR or any experimental work to validate the food adulteration in a very
short time, as little as 30 minutes. Figure 4 illustrates the process flow and mechanism of the
DNAFoil technique.
Herein, this technique can detect food adulteration by following these five main steps（Sheikha, 2019）: initially, the provided barrel to break, lyses, extract, neutralize, and stabilize
DNA in the food sample. Subsequently, the DNA target amplifies by transferring one drop of theextracted DNA（step 1）to the reaction tube, which is then placed in the water. The DNA target
will amplify and make multiple copies using enzymes and specific primers identified. Finally, immerse the revealing strip after waiting about 30 minutes to detect any traces. The target DNA
will be transferred via capillary force to the detection surface of the test strip during the final
stages. The target DNA fragments will be captured in a band on the test strip. These proceduresshow the versatile, potent, and rapid technology of DNAFoil.

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Fig. 4

Process flow and mechanism of DNAFoil technique（Sheikha, 2019）

This DNAFoil kit technology has successfully detected pork adulteration in beef at lower
levels, which was reported in Meat and Livestock Australia Limited in January 2018. The
DNAFoil validity has been confirmed using qPCR. Aronoff et al.（2018）demonstrated the reproducibility data to detect the vegetal material in milk products using qPCR. This finding indicatesthis DNAFoil kit technology is a rapid and reliable method to validate product content. It canalso be used to identify the characteristics in minimal time without needing any lab equipment,
technicians, or scientists.
6.

Conclusion
The development of food integrity management system within food supply chain is import-

ant to create trust and brand integrity among stakeholders. This can be achieved through a few
contexts by ensuring :（i）safety, quality, and authenticity of food products,（ii）reliable labelling,
and（iii）effective management of provenance such as halal status, vegan, or organic product. In
the sense of the first context, adulteration and authenticity is a key issue that has been gaining
increasing attention by all players including industry, government, academia, and consumers inrecent years.
A tremendous improvement on the analytical methodologies to investigate the food productintegrity especially in detecting or analyzing the adulterants are seriously in need due to thecountless food scandals have evolved. This is important to gain the consumer confidence and protecting consumer rights by guarantee the quality, safety, and authenticity of the food product aswell as processing methods according to the standard operating procedures.
For this reason, this chapter reviewed the food integrity in the perspective of food scienceand technology on the safety, quality, and authenticity of the food product within the wider foodsupply chain. With an increasing crises and food industry scandals, a rapid analytical technique
has been developed to carter with this issue. Herein, the available analytical methods particularly

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for chemical/biochemical and DNA-molecular technique have been reviewed. This review indicates a significant enhancement of the analytical methods development to mitigate the adulteration or authenticity issues by promoting several advantages such as high efficacy, rapid, and minimal cost. Nevertheless, there still need a continuous research and development to improve thelimitation of the techniques.
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