# US 2018/0023032 A1 · Solventless winterization of microbial oil > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2018/0023032 A1 - Document ID: US20180023032A1 - Status: Published application - Kind: A1 - Application number: 15/655,433 - Filed: 2017-07-20 - Published: 2018-01-25 - Earliest claimed: 2016-07-20 - Assignees: Mara Renewables Corporation - Applicants: Mara Renewables Corporation - Inventors: Xuan Jiang; Dorothy Dennis; Roberto E. Armenta - Web page: https://patentgrep.com/patent/US20180023032A1 ## CPC - C11B 7/0075: {by differences of melting or solidifying points} - C11B 3/006: {by extraction} - C11B: PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES (drying-oils C09F) ## Abstract Provided herein are methods for winterizing oil. The methods include heating the oil to a first temperature and maintaining the oil at the first temperature for a first period of time; reducing the first temperature of the oil after the first period of time to a second temperature over a second period of time, wherein reducing the first temperature produces a first solid fraction and first liquid fraction of the oil; removing the first solid fraction from the oil; reducing the second temperature of the first liquid fraction of the oil over a third period of time to a third temperature, wherein reducing the second temperature of the oil produces a second solid fraction and second liquid fraction of the oil; removing the second solid fraction from the oil; and recovering the second liquid fraction of the oil. ## Claims ### Claim 1 A method for winterizing oil comprising the steps of: (a) providing an oil; (b) heating the oil to a first temperature and maintaining the oil at the first temperature for a first period of time; (c) reducing the first temperature of the oil after the first period of time to a second temperature over a second period of time, wherein reducing the first temperature produces a first solid fraction and first liquid fraction of the oil; (d) removing the first solid fraction from the oil; (e) reducing the second temperature of the first liquid fraction of the oil over a third period of time to a third temperature, wherein reducing the second temperature of the oil produces a second solid fraction and second liquid fraction of the oil, and wherein the oil is not heated between the steps of reducing the first temperature and reducing the second temperature; (f) removing the second solid fraction from the oil; and (g) recovering the second liquid fraction of the oil. ### Claim 2 (depends on 1) The method of claim 1, wherein the method is carried out in the absence of solvents. ### Claim 3 (depends on 1) The method of claim 1, wherein the first temperature is above the melting point of the oil. ### Claim 4 (depends on 1) The method of claim 1, wherein the first temperature is from 25° C. to 65° C. ### Claim 5 (depends on 4) The method of claim 4, wherein the first temperature is from 40° C. to 65° C. ### Claim 6 (depends on 1) The method of claim 1, wherein the oil is maintained at the first temperature for 5 to 60 minutes prior to the reducing step. ### Claim 7 (depends on 1) The method of claim 1, wherein the first temperature is reduced by 0.5 to 2 degrees per hour over the second period of time to the second temperature. ### Claim 8 (depends on 1) The method of claim 1, wherein the first temperature is reduced by 0.5 to 2 degrees per minute over the second period of time to the second temperature. ### Claim 9 (depends on 1) The method of claim 1, wherein the second period of time is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours. ### Claim 10 (depends on 1) The method of claim 1, wherein the oil is mixed during the second period of time. ### Claim 11 (depends on 10) The method of claim 10, wherein the mixing comprises a speed of 50 to 200 rpm. ### Claim 12 (depends on 1) The method of claim 1, wherein the second temperature is the cloud point of the oil. ### Claim 13 (depends on 1) The method of claim 1, wherein the second temperature is between 10° C. to 20° C. ### Claim 14 (depends on 1) The method of claim 1, wherein the second temperature is between 20° C. to 30° C. ### Claim 15 (depends on 1) The method of claim 1, wherein the oil is maintained at the second temperature for 5 to 20 minutes. ### Claim 16 (depends on 1) The method of claim 1, wherein the second temperature is reduced by 0.5 to 2 degrees per hour over the third period of time to the third temperature. ### Claim 17 (depends on 1) The method of claim 1, wherein the second temperature is reduced by 0.5 to 2 degrees per minute over the third period of time to the third temperature. ### Claim 18 (depends on 1) The method of claim 1, wherein the third period of time is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours. ### Claim 19 (depends on 1) The method of claim 1, wherein the third temperature is room temperature. ### Claim 20 (depends on 1) The method of claim 1, wherein the third temperature is 4° C. ### Claim 21 (depends on 1) The method of claim 1, wherein the method further comprises reducing the third temperature of the second liquid fraction of the oil over a fourth period of time to a fourth temperature, wherein reducing the third temperature of the oil produces a third solid fraction and third liquid fraction of the oil. ### Claim 22 (depends on 21) The method of claim 21, wherein the method further comprises removing the third solid fraction of the oil. ### Claim 23 (depends on 21) The method of claim 21, wherein the method further comprises recovering the third liquid fraction of the oil. ### Claim 24 (depends on 21) The method of claim 21, wherein the fourth temperature is room temperature. ### Claim 25 (depends on 21) The method of claim 21, wherein the fourth temperature is 4° C. ### Claim 26 (depends on 1) The method of claim 1, wherein 80% or more of the oil is clear at room temperature. ### Claim 27 (depends on 1) The method of claim 1, wherein the oil comprises one or more polyunsaturated fatty acids. ### Claim 28 (depends on 27) The method of claim 27, wherein the polyunsaturated fatty acid is docosahexaenoic acid (DHA). ### Claim 29 (depends on 28) The method of claim 28, wherein the oil comprises 40% or more DHA. ### Claim 30 (depends on 1) The method of claim 1, wherein the oil is derived from a population of microorganisms. ### Claim 31 (depends on 30) The method of claim 30, wherein the population of microorganisms is selected from the group consisting of algae, fungi, bacteria and protists. ### Claim 32 (depends on 30) The method of claim 30, wherein the population of microorganisms is selected from the genus Oblongichytrium, Aurantiochytrium Thraustochytrium, and Schizochytrium or any mixture thereof. ### Claim 33 (depends on 30) The method of claim 30, wherein the population of microorganisms is a Thraustochytrium sp. deposited as ATCC Accession No. PTA-6245.