Methyl oleate

Culture medium and supplements were purchased from Life Technologies. Palmitate, oleate, elaidate, vaccenate, FA free bovine serum albumin, trans-vaccenic acid methyl ester, methyl oleate, methyl Palmitate, methyl palmitoleate methyl stearate, 1,3-diheptadecanoyl-glycerol (d5) (>99%), 1,2,3-triheptadecanoyl-glycerol (>99%), C17:0 ceramide (>99%), n-hexane, methanol (gradient grade) and isopropanol (gradient grade) and boron trifluoride–methanol solution were purchased from Merck. All other chemicals used in this study were of analytical grade. All experiments and measurements were carried out by using Millipore ultrapure water.

After the extraction of pure biodiesel from the rotary evaporator under optimal conditions, a considerable amount of biodiesel is characterized using ¹H and ¹³C nuclear magnetic resonance spectroscopy to confirm the formation of biodiesel (methyl oleate). ¹H and ¹³C nuclear magnetic resonance spectroscopy was carried out in a Jeol instrument having model number ECZ500R/S1 having a frequency ranging from 10 to 535 MHz. The conversion of oleic acid into methyl oleate is obtained using Eq. (2). $$Conversion\left(\%\right)=\frac{{2A}_{\mathit{OMe}}}{{3A}_{CH2}}\times 100$$
In this equation (Eq. 1), A_OMe stands for the integration of the OMe groups in FAME, and A_CH2 stands for the total integration of the –CH₂ (OA + FAME).
In order to verify the findings of the NMR investigation, a GC (gas chromatography) analysis was performed. GC was carried out using an Agilent model 8890 equipped with split/splitless injectors, polar columns (HP-5), and a UI Agilent column having diameter 30 m × 0.320 mm. The temperature of the oven was raised from 300 to 400 °C where the injection temperature was 300 °C. An external calibration curve has been prepared by methyl oleate (purchased from Merck) to quantify the peaks.