ADVANCES IN VEGETABLE OIL AUTHENTICATION BY DNA-BASED MARKERS

ABSTRACT:The suitability of DNA markers in providing unequivocal identifiers for authentication and traceability of food has been a subject of an increasing number of reports. Even in complex food matrices such as vegetable oils, the use of molecular markers as diagnostic tools has been exploited. Considering the wide variety of vegetable oils available for consumers and the differences in prices, especially among premium olive oil and other oils, species adulteration leading to economic losses and loss of consumer confidence can arise. In this review, the advances of DNA extraction protocols are emphasized as a crucial step to overcome. Specific identification of several plant oils as potential adulterants of olive oil has been a subject of very recent progresses. When the oilseed crops are the source for vegetable oil production, additional concerns due to the presence of genetically modified organisms have prompted to further improvements in DNA analysis. In the specific case of olive oil, the use of genetic markers has provided analytical tools to assess authenticity regarding cultivar identification as independent markers from environmental fluctuations. INTRODUCTION There is a wide variety of vegetable oils available to consumers and their prices mostly depend on the oils quality and purity. Unfortunately, some counterfeit vegetable oils adulterated by

illegal operators are often being sold for profiteering. Species adulteration of vegetable oil will lead to economic losses and loss of consumer confidence, and may even negatively impact legitimate operators. Therefore, a reliable method of species authentication for vegetable oils is desirable. Currently there are two major methods for vegetable oil species authentication. The first is the conventional method of chemical constituent analysis to identify the specific vegetable oil based on species-specific chemical composition. Analytical techniques such as proton transfer reaction mass spectrometry (PTR-MS), nuclear magnetic resonance spectroscopy (NMR), high performance liquid chromatography and gas chromatography are commonly used to analyze olive oil. However, the chemical composition of oil may differ among different seasons and growing areas. Moreover, it was found that no single known parameter could detect the presence of other oils in olive oil, if their concentration is lower than or equal to 5% (Christopoulou, Lazaraki, Komaitis, & Kaselimis, 2004). Consequently, chemical analyses may not be sufficient for oil authentication (Gimnez, Pistn, Martn, & Atienza, 2010). The second method is based on DNA genetic identification and involves the use of polymerase chain reaction (PCR) and molecular markers. PCR

technology has been proven to be an invaluable method for species identification via amplification of species-specific genes, which can be employed to identify species composition in food Real-time PCR is frequently used for quantitative and qualitative analysis of species-specific genes, such as the detection of genetically modified soybean DNA in refined vegetable oils Although PCR is a rapid and sensitive technique, it is expensive and prone to generating false positive signals Molecular marker methods such as random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), and amplified fragment length polymorphism (AFLP) techniques have been developed and optimized. DNA marker methods require that the samples are derived from a single cultivar, but also rely on the accessibility of sufficient reference cultivars. Evidently, these two limitations are important disadvantages. TRACEABILITY OF VEGETABLE OILS. Adulteration is generally motivated by maximizing profit by adding a cheaper ingredient. The growing interest in the authenticity of foods and food products requires reliable verification methods because the properties of foods from different origins can also be different. Furthermore, traceability is required more and more by consumers and government organizations

because it is a significant component of overall food safety (Anklam and Battaglia, 2001). The methods used for the verification of food authenticity and traceability have to be suitable for the prevention of deliberate or accidental mislabelling. They are important for commercial reasons and play a considerable role in the assurance of public health. ANALYSIS OF MACRO- AND MICROCOMPONENTS OF OILS OF VARIOUS ORIGIN BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) AND GAS CHROMATOGRAPHY (GC) Plant oils contain many components, such as triacyland diacyl-glycerols (TAGs and DAGs) of various saturated and unsaturated fatty acids as main components, and tocopherols, phytosterols, phospholipids, free fatty acids, waxes as minor ingredients. The composition of the oils (triglyceride profile) is a characteristic of the plant species and to a lesser extent for the origin of the oil. As the macroand microcomponents of oils are semi-volatile, both HPLC and GC techniques can be employed for their measurement. Consequently, a considerable number of HPLC and GC methods have been developed for the determination of the composition of various oils. As the

evaluation methods used for the authenticity and traceability tests of oils include a high number of variables (e.g. composition of TAGs) the applications of various multivariate mathematical and statistical methods greatly facilitate the evaluation of the results. The chromatographic techniques applied for the authenticity test of vegetable oils have been previously collected and critically evaluated (Aparicio and Aparicio-Ruiz, 2000).