WO2018195422A1 Worldwide applications  2018 WO US EP 2021 US 

 EXAMPLE 1
[0050] Example 1. Extraction of ethyl ferulate and ethyl coumarate from corn fiber. In this example, 1.005 g of corn fiber that was previously dried at 100 °C for 24 hr was loaded into a 100 mL stirred batch reactor (Parr Instruments 2430) with 20 mL 200 proof anhydrous ethanol. The reactor was sealed and purged with 99.999% hydrogen (Airgas Hy UHP) four times by pressurizing the reactor to 17.2 bar and subsequently venting the pressure to ca. 3 bar. The reactor was pressurized to a final pressure of 34.5 bar. The temperature was increased to 200 °C at a ramp rate of 300 °C hr"1 while the reactor was stirred at 200 rpm. The temperature was held for 12 hours before returning to room temperature. After reaction, the solvent and solids were filtered through filter paper (particle retention > 11 µp?) and rinsed with methanol to a total volume of 50 mL. The ethyl ferulate content of the liquid was analyzed by HPLC Method 1, as shown in FIG. 5. Ethyl coumarate and ethyl ferulate eluted at ca. 30.4 and 31.6 min, respectively. The yield of ethyl ferulate was determined to be 1.0% with respect to the total dry mass of corn fiber (assuming 8% moisture content of the biomass, dry mass = 0.92 * 1.005 g = 0.925 g).

EXAMPLE 2
[0051] Example 2. Extraction of methyl ferulate and methyl coumarate from miscanthus. In this example, 255.2 g of miscanthus was loaded into a 7.5 L stirred batch reactor (Parr Instruments 4550) with 4 L methanol. The reactor was sealed and purged with 99.999%) argon (Airgas Ar UHP) four times by pressurizing the reactor to 7 bar and subsequently venting the pressure to ca. 2 bar. The reactor was pressurized to a final pressure of 7.6 bar. The temperature was increased to 200 °C at a ramp rate of 150 °C hr"1 while the reactor was stirred at 200 rpm. The temperature was held for 12 hours before returning to room temperature. After reaction, the solvent and solids were filtered through a nylon bag (particle retention > 75 µp?) and rinsed with methanol to a total volume of 3.2 L. The methyl ferulate and coumarate content of the liquid was analyzed by HPLC Method 1, as shown in FIG. 6. Methyl coumarate and methyl ferulate eluted at ca. 25.5 and 26.5 min, respectively. The yield of methyl ferulate and methyl coumarate were determined to be 0.5% and 3.9%, respectively, with respect to the total dry mass of miscanthus (assuming 8% moisture content of the biomass, dry mass = 0.92 * 255.2 g = 234.8 g).

XAMPLE 3
[0052] Example 3. Extraction of ethyl ferulate from corn fiber at 170 °C. In this example, 7.5 kg of corn fiber previously dried at 90 °C for 24 hr under vacuum was loaded into a 1 13.6 L stirred batch reactor with 60 L 200 proof anhydrous ethanol and 144 g NaOH. The reactor was sealed and purged with nitrogen 3 times by pressurizing the reactor to 7 bar and subsequently venting the pressure to ca. 2 bar. The reactor was filled with nitrogen to a final pressure of 1 bar. The temperature was increased to 170 °C at a ramp rate of 225 °C hr"1 while the reactor was stirred at 600 rpm. The temperature was held for 12 hours before cooling to room temperature. After reaction, the solvent and the solids were filtered through a nylon filter bag and rinsed with 19 L ethanol. The ethanol oil was concentrated with a rotary evaporator to 3.1 kg. The ethyl ferulate content of the oil was analyzed by HPLC Method 1, as shown in FIG. 7. Ethyl coumarate and ethyl ferulate eluted at ca. 31.2 and 32.4 min, respectively. The yield of ethyl ferulate was determined to be 1.5% with respect to the total dry biomass (assumed to be 7.5 kg in this case because the fiber was dried immediately before processing).

EXAMPLE 4
[0053] Example 4. Extraction of ethyl ferulate from corn fiber at 145 °C. In this example, 1.0 g of corn fiber that was previously dried at 100 °C for 24 hr was loaded into a 100 mL stirred batch reactor (Parr Instruments 2430) with 20 mL 200 proof anhydrous ethanol. To the reaction was added 32 mg NaOH. The reactor was sealed and purged with 99.999%) argon (Airgas Ar UHP) four times by pressurizing the reactor to 17.2 bar and subsequently venting the pressure to ca. 3 bar. The reactor was pressurized to a final pressure of 3 bar with Argon gas. The temperature was increased to 145 °C at a ramp rate of 300 °C hr"1 while the reactor was stirred at 200 rpm. The temperature was held at 145 °C for 12 hours before returning to room temperature. After reaction, the solvent and solids were filtered through filter paper (particle retention > 1 1 µp?) and rinsed with methanol to a total volume of 50 mL. The ethyl ferulate content of the liquid was analyzed by HPLC Method 2, as shown in FIG. 8. Ethyl coumarate and ethyl ferulate eluted at ca. 30.4 and 31.6 min, respectively. The yield of ethyl ferulate was determined to be 1.66% with respect to the total dry mass of corn fiber (assuming 8%> moisture content of the biomass, dry mass = 0.92 * 1.0 g = 0.92 g).

 EXAMPLE 5
[0054] Example 5. Extraction of ethyl ferulate from corn fiber at 120 °C. In this example, 1.0 g of corn fiber that was previously dried at 100 °C for 24 hr was loaded into a 100 mL stirred batch reactor (Parr Instruments 2430) with 20 mL 200 proof anhydrous ethanol. To the reaction was added 32 mg NaOH. The reactor was sealed and purged with 99.999% argon (Airgas Ar UHP) four times by pressurizing the reactor to 17.2 bar and subsequently venting the pressure to ca. 3 bar. The reactor was pressurized to a final pressure of 3 bar with Argon gas. The temperature was increased to 120 °C at a ramp rate of 300 °C hr"1 while the reactor was stirred at 200 rpm. The temperature was held at 120 °C for 12 hours before returning to room temperature. After reaction, the solvent and solids were filtered through filter paper (particle retention > 11 µp?) and rinsed with methanol to a total volume of 50 mL. The ethyl ferulate content of the liquid was analyzed by HPLC Method 2, as shown in FIG. 9. Ethyl coumarate and ethyl ferulate eluted at ca. 30.4 and 31.6 min, respectively. The yield of ethyl ferulate was determined to be 1.52% with respect to the total dry mass of corn fiber (assuming 8% moisture content of the biomass, dry mass = 0.92 * 1.0 g = 0.92 g).
[0055]