1. Introduction
The interest in dietary fat is widespread, and fatty compound analysis is a basic requirement in testing food material [1]. Animal fats tend to have a larger proportion of long chain saturated acids and are solids at room temperature [2]. The different kinds of FA are saturated fatty acid (SFA), monounsaturated (MUFA), polyunsaturated (PUFA) and trans (TFA) [3]. The most widely used accurate and repeatable method for FA analysis is GC equipped with flame ionization detector (FID) [4].
TFAs are defined as unsaturated FA (UFA) with at least a double bond in the trans configuration resulting in a more rigid molecule close to a SFA [5]. Fats from plant sources contain a higher proportion of unsaturated acids and are often liquids at room temperature due to hydrogen bonding [6]. Industrial hydrogenation which is performed to improved texture, stability and other economically desirable properties is the major cause of TFA creation [7]. Also, microbial transformation of UFA in ruminants, heating and frying above 180˚ and deodorization of edible oil are the other causes of TFA creation [8]. Hydrogenation is a process that reduces the relative unsaturation of the oils and promotes geometric and positional isomeration [9]. Several clinical studies have shown that a TFA diet increase unfavorable LDL (low density lipoprotein)/HDL (high density lipoprotein) ratio and lipoprotein (a) and plasma triglyceride levels, which are independently associated with the increase risk of coronary heart disease (CHD) [10]. The risk of ischemic heart disease increases 25% by daily intake of TFA as little as 5 g [8]. Moreover, TFA intake promotes inflammation, increases body mass index, raises C-reactive protein, causes endothelial dysfunction and relates to the risk of diabetes. Replacing saturated or trans fat with polyunsaturated or monounsaturated fat is favorable in lowering serum cholesterol and reducing risk of CAD (Cardiac attack death) [11].
In Iran, TFA accounts 4.2% of all calories consumed by Iranians, which is much higher than US and many European populations do [12]. Between 11% and 39% of coronary heart disease referring to consumption of TFA and also, elimination of TFA in hydrogenated vegetable oils (HVOs) might be prevented 8% - 39% of cardiovascular disease events among Iranians [13]. In US populations TFA account for 2% - 3% of total energy intake [14].
In spite of the TFA disadvantages, limited data on the TFA contents of Iranian foods are available and it seems that the amount of TFA in edible oil requires more attention in developing countries [15].
Therefore, the aim of this study was to determine and compare TFA content in the branded liquid frying and solid oils that are commonly consumed by Iranian and also comparison of the achieved amount with approved standard in Iran and the other countries.
2. Materials and Methods
2.1. Sampling
Eight brands of liquid frying oils and four brands of solid oils that are commonly consumed in Iran were selected and each brand was coded with a letter (A, B, C, D, and…). Lot numbers were checked to ensure that each unit belonged to a different lot. All oil samples were stored, labeled and analyzed before expiry dates. Samples were selected to include the major manufacturers of the oils in Iran. The composition of studied oils was mixture of palm oil, soybean oil, canola oil, and sunflower oil. However solid oils contain more percentage of saturated fat.
2.2. Reagents and Standards
All solvents and reagents were used of analytical grade. Methanol, n-Hexane, sodium hydroxide, methanolic boron trifluoride and sodium chloride were purchased from Merck, (Darmstadt, Germany).
The certified standard mixture of 37 component FA methyl ester C4-C24 was purchased from (Accuastandard, USA), also linoleic and linolenic acid methyl ester isomer were purchased from (Sigma, USA).
2.3. Sample Preparation
Methyl-esterification of samples used in the analyses was performed by BF3-MeOH method according to AOCS (American oil chemist society) [16]. In to a 125 mL flask, one g of oil was weighted and added ten mL of 0.5 (N) methanolic sodium hydroxide solutions, the mixture was heated at 100˚C for 10 min, then 12 mL BF3-MeOH reagent was added, and heated for extra 2 min. After cooling five mL of n-hexane was added. The reaction was stopped by adding 15 mL saturated solution of NaCl and shaking the flask for about 15 seconds. One mL of the upper organic phase was selected and anhydrous Na2SO4 was added. The sample solution was injected to the gas chromatography (GC) after filtering through 0.22 µm disposable syringe filter.
2.4. GC-FID Analysis
The GC-FID analyses were performed on an Agilent model 7890 GC instrument equipped with a flame ionization detector. A highly polar capillary column (100 m × 0.25 mm i.d × 0.25 µm film thickness) of HP-88 (Agilent, USA) was used to separate the FAMEs. Nitrogen was used as the carrier gas at a flow rate of 1.0 mL/min. A split ratio of 100:1 was used and 1 µL of the sample was injected into the GC for analysis. The following oven temperature program was used: 180˚C for 30 min, then increase to 200˚C at a rate of 1.5˚C/min and kept at 200˚C for 30 min. The temperature was set at 220˚C for the injector and at 250˚C for the detector. The samples were analyzed triplicate and the results were expressed as mean values ± standard deviation (SD).
2.5. Data Analysis
Data were analyzed using statistical package for social sciences, version 16 (SPSS Inc., Chicago, IL, USA). Three independent oil samples of each cultivar were analyzed three times. Data are expressed as mean _ SDs. T-test was used for determining the differences between means inliquid frying oils and the solid oils. Statistical significance was set at p < 0.05.
3. Results and Discussion
3.1. Liquid Frying Oils
The FA compositions of various brands of liquid frying oils in Iran are presented in Table 1. The mean of SFA in the brands ranged from 11.67% in C to 36.12% in A. Among the SFA palmitic acid presented the highest value ranging from 6.86% in C to 30.34% in D. High oleic oils (C18:1c) have better oxidative stability in deep frying applications and extended shelf life and reduced LDL [17]. Group A had maximum content of oleic (36.20%).
Table 1 shows that UFA ranged from 27.36% in A to 54.24% in C. Linoleic and linolenic acid are the most important PUFA. As nutritional point of view the amount of PUFA in oils is very important. Linoleic and linolenic have useful influence on human health and improve cardiovascular function. They also have positive effect on lipid profile [7]. Despite of these positive effects, high levels of multiple double bond FA increase oxidation oil sensitivity in liquid frying oil [17]. Therefore, high linolenic content was found in group C (3.85%) due to more oxidation tendency. According to the acceptable range of linolenic in Iran, standard reference and also Wolff report [18] on only C group with 3.85% mean percentage was