#208

Extraction of Oil from Oilseeds by Pressurized Solvent Extraction (PSE)

Introduction
Pressurized solvent extraction is a new technique that reduces solvent consumption and sample preparation time. Solvent is pumped into an extraction vessel containing the sample and is heated and pressurized. The pressurized solvent at high temperature accelerates the extraction process by increasing the solubility of the analyte in the solvent and also increasing the kinetic rate of desorption of the analyte from the sample matrix. Traditionally, oil is removed from oilseeds such as rapeseed, soy and cottonseed using tested methods such as the American Oil Chemists Society Method Am-2-93. This is a solvent-based method that employs a Soxhlet apparatus to extract oils from oil seeds. The method must be carried out in three stages requiring the apparatus to be disassembled after the first two of three extractions, and the sample reground in mortar before reassembly. The entire process may take as long as 20 hours. This application describes a procedure for the rapid extraction of oil from oilseeds using pressurized solvent extraction for significant timesavings and reduced solvent consumption.

Equipment
Applied Separations one PSE Pressurized Solvent Extractor 11 mL Extraction Vessel-Cat.#10625 Microwave Oven for sample drying Mortars (90mm o.d. Coors #60316) and pestles (Coors #60317)

#208

Solvents and Materials

Petroleum ether (b.p. 35 - 60C) or Hexane (ACS grade) Nitrogen high quality grade S/S Frits (10 micron)- Cat. #10710 Collection Vials (60mL for extract collection)-Cat.#10650 Spe-ed Matrix-Cat.#7950 Cellulose Filter Disk-Cat. #10711 Ottawa SandCat. #10548

Summary of Method

1. Prepare Sample

2. Load Sample

3. Run Sample

4. Collect Extract

Procedure
Sample Preparation Oilseed samples must be ground to a particle size of 3 mm or less before proceeding with this method. Failure to do so may result in inconsistent results. Place 2 g Spe-ed Matrix in a weighing dish. Accurately weigh 1 to 1.5 g (to 0.1 mg) of the oilseed sample onto the Spe-ed Matrix in the weighing dish. Pour this material carefully into the mortar and grind until uniform with the pestle. Scrape all particles from pestle into mortar then set the pestle aside.

#208

Place only the mortar in a microwave oven and heat for four minutes at a high setting. When mortar cools, regrind mixture with pestle until smooth. Load Sample Prepare the extraction vessel(s) for analysis by placing a filter disk in the bottom opening followed by a 10m frit, and secure them in place with the retaining nut. Place adaptor and funnel in top opening. Pour the sample mixture through a funnel into the extraction vessel. Add an additional gram of Spe-ed Matrix to the mortar and grind to sweep up any traces of residual sample. Transfer this additional material to the extraction vessel and tap the vessel on the bench top to settle and compact the contents. Add Ottawa sand to within 1 cm of the top of the vessels interior flange (see illustration on page 4-3 of Users Manual). Transfer vessel to oven well of one PSE and close cover. Ensure that the pump is primed and that the extraction solvent is petroleum ether or hexane. Attach tared collection vial to needle port and program the instrument using the following parameters: Extraction Conditions Program the following extraction parameters on the one PSE Program A Mode 11 mL vessel Solvent: Hexane Temperature: 110 C Pressure: 100 Bar Cycles: 3 Static: 25 minutes Pause: N=0 Flush: Solvent/gas/repeat flush:20 sec/2min/0 At the end of the extraction, remove collection vial from the needle port and place vial in an evaporation apparatus. Remove solvent using low heat under a gentle stream of nitrogen. Dry vial in oven set at 100 C for 1 hour. Weigh dried vial, calculate the percent recovery and report result.

#208

Example: Extraction of Oil from Rapeseed The one PSE was used for the extraction of a Rapeseed sample, which was also extracted in triplicate using the official AOCS Method Am 2-93. The results of the two determinations types are shown in the following table: Percent Recovery of Oil from Rapeseeds by one PSE vs Soxhlet % Recovery one PSE n AM 2-93 45.00 0.29 6 44.87 1.27 (0.64) (2.83) n 3

Mean sd (RSD)

Six determinations were performed on the rapeseed sample by the one PSE method while 3 determinations were carried by the official method. The sample sizes for the one PSE averaged 1.5 g, while those for the Soxhlet method were larger and averaged 3.5 g. In spite of the larger number of determinations performed on the one PSE, the RSDs obtained by this technique were significantly lower (0.64 vs. 2.83) than by the official method. The mean for the one PSE samples was also slightly higher than that for the Soxhlet samples. Moreover, these six determinations could be carried out in far less time than the three obtained by AM-2-93 and with significantly less solvent: 30 mL for the one PSE vs. 250 300 mL for the Soxhlet.

#208

Safety
The use of organic solvents, elevated temperatures, and high pressures present potential safety concerns in the laboratory. Common sense laboratory practices can be employed to minimize these concerns. However, the following sections describe additional steps that should be taken. Extraction vessels in the one PSE oven are hot enough to burn unprotected skin. Allow the vessels to cool before removing them from the oven, or use appropriate protective equipment (e.g. insulated gloves or tongs) as recommended by the manufacturer. During the gas purge step, some solvent vapors may exit through a vent port in the instrument. Connect this port to a fume hood or other means to prevent release of solvent vapors to the laboratory atmosphere. This precaution also applies to the removal of post extraction solvent from the collected extract.