D potassium hydroxide, and sodium hydroxide) had been of analytical reagent grade and bought from Systerm (Systerm, Malaysia) except for n-hexane, which was of greater purity (Systerm, mGluR5 Antagonist medchemexpress Malaysia, for GC, 99 ). The esterifying agent P2Y14 Receptor Agonist Purity & Documentation TMS-DM (2 M) in n-hexane was bought from Sigma (Sigma-Aldrich, Germany). two.2. Food Samples. Eight industrial meals products had been made use of for analysis and comparison in this study. The samples included unique bakery and fast-food items, which include crackers, bread with filling, cakes, wafers, cookies, and biscuits, as these merchandise primarily include FAs and TFAs. The samples have been bought from many Malaysian nearby supermarkets, such as national and imported brands, and all of these samples were coded with a letter (from A to H). two.3. Sample Preparation and Lipid Extraction. Every single sample was ground and placed in an oven at 50 C until full dryness ahead of evaluation. The total lipids had been extracted making use of the Soxhlet System for cereal fats [28]. About 10 g of homogenized sample was weighed into a cellulose extraction cartridge, plus the Soxhlet apparatus containing the cartridge was fitted to a distillation flask containing 150 mL of nhexane with (50 ppm) butylated hydroxytoluene (BHT) and a few antibumping granules. After 3 hours, the mixture was dried with Na2 SO4 and filtered by means of fluted filter paper. The oil was recovered immediately after stripping the solvent in a rotary evaporator. Finally, the extracted lipids had been dried beneath nitrogen (N2 ), weighed,and stored at -20 C until evaluation. 2.four. Preparation of Fatty Acid Methyl Esters (FAMEs). After Soxhlet extraction, all lipid extracts had been methylated and converted into FAMEs employing two distinct methylation solutions. About 0.15 g of every single fat extract (in triplicate) was transferred to a screw-cap test tube (ten mL), and 1 mL ofThe Scientific Planet JournalOCH2 OC O C O CR R(a)CHO (b) KOCH3 /HCl system NaOCH3 CH3 OH 60 C OCH3 O CH2 ORKOCH3 /HCl strategy KOH CH3 OH 70 CTriacylglycerolC OR(FAME)HOCH3 H3 C Si N2 CH3 TMS-DM 50 C Methanol: toluene 2CR(FFA) HCl 1.0 MOCH3 OCR(FAME)Figure 1: Diagram for the procedures with the technique (a) (KOCH3 /HCl) and system (b) (TMS-DM).a remedy containing ten mg/5 mL (C15:0) in methanol was added as an IS. The mixtures have been lowered to dryness beneath nitrogen (N2 ) before derivatization working with two distinct methodologies (Figure 1), and the procedures were performed as described inside the following sections. 2.four.1. Base-Catalyzed Followed by the Acid-Catalyzed System (KOCH3 /HCl). The mixtures were redissolved in 2 mL of nhexane, and 1 mL of two M methanolic KOH option was added for the samples. The tubes had been capped and vigorously shaken for 30 s and boiled for two min inside a water bath at 70 C. Then, 1.two mL of HCl (1.0 M) was added plus the remedy was gently stirred. Just after phase separation, 1 mL of n-hexane was added. The upper phase containing the FAMEs was transferred into an evaluation vial, and 1.0 L with the solution was injected in to the GC-FID. two.four.2. Base-Catalyzed Method Followed by TMS-DM. The mixtures had been redissolved in two mL of n-hexane, and 1 mL of 2 M NaOCH3 was added. The content material was placed inside a water bath at 60 C for five min. Drops of concentrated glacial acetic acid have been added to each tube to neutralize the NaOH. The samples had been decreased to dryness below N2 and dissolved in 1 mL of methanol : toluene (two : 1 vol.). Subsequent, TMS-DM was added in a molar excess of two M in n-hexane (one hundred L) at 50 C for 10 min devoid of capping the tubes.