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Figure 3: Solubility of oleate species in scCO2. Data shown as a compilation [39] across density from a range of temperatures and pressures Phase behavior. This research seeks to identify a reaction pathway (temperature, pressure, concentration) that allows for conversion of starting material to product with the desired phase behavior as seen for the pure methanol system at temperatures above 225 ̊C [40]. Previous work on the use of methanol as a solvent for TG transesterification employed supercritical temperatures and pressures, not so much because supercritical conditions were desired, but rather because the high temperatures (and consequently high pressures) were needed to create a single phase of methanol and non-polar TG. In our system we propose to employ a CO2 and methanol mixture to allow for transesterification at more ambient temperatures. The fundamental understanding of the system’s phase behavior and phase conditions for efficient reaction/extraction is explored. MATERIALS AND METHODS Algal growth. Algal cultures were grown in a small-scale reactor setup in media as recommended by [42]. 14 h/day of illumination will be provided with an artificial light source consisting of two fluorescent light bulbs per reactor vessel. The reactors were sparged with house air with an addition of 2% CO2. The growth of each algae species was monitored by UV/Visible spectroscopy with cell density being correlated with absorbance at 610 nm as measured by direct cell count. 40 mL aliquots of algae were placed into 50mL Falcon tubes, centrifuged (12,000 rpm, 20 mins, 4 ̊C), the supernatant decanted, and the algae frozen at -20 ̊C until use. The wet algae mixture contains 4% solids with a total sample volume of 1 mL. The algae will then either be used wet or lyophilized to dryness and kept in a desiccator. Extraction. Conventional solvent extraction was conducted with slight modifications to Bligh and Dyer[11] using 2:1 chloroform:methanol. Extraction using scCO2 was performed using a supercritical fluid extractor (SFT-100, Supercritical Fluid Technologies). Samples were added to the extraction chamber, run continuously for one hour at specified pressure and temperature, and the flow through was collected at a rate of 4 SCFH of gaseous flow (~0.2 kg CO2 per run). Analysis. Using liquid chromatography-mass spectrometry (LC-MS) with atmospheric pressure chemical ionization (APCI), TG, DG, MG, glycerol, and FAME concentrations were measured. Mass spectrometry allows for the compounds to be identified by both retention time and molecular weight. Heptane with 0.1% acetic acid was used as the elution solvent to successfully separate FAME standards as well as several triolein, diolein, and monoolein [43]. The column used is a normal phase HILIC column (Waters) at a flow rate of 0.25 mL/min. Mass spectral settings were optimized for FAME with short, medium, and long chain lengths at capillary voltages of 75, and 110V and RF loadings of 80%, and 95% respectively. Standards are used to verify the presence and quantify each substrate. Phase Behavior Determination. The phase behavior of the reaction mixtures was observed in a variable volume, high pressure view cell (D.B. Robinson and Associates; Edmonton, Alberta, Canada) as performed in previous works [44, 45]. Cloud point curves were measured isothermally. Methanol and substrate was added to the cell at a pre- determined ratio and the system sealed. Once the desired temperature is achieved, a measured quantity of CO2 at isPDF Image | One-Pot Algal Biodiesel Production in Supercritical CO2
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