Process for collecting minor components with double pass and using enzyme

Abstract

The present invention relates to a process for obtaining minor components from vegetable oils, wherein the minor components of interest are squalene, tocopherols, and optionally tocotrienols. The process is comprising, an enzymatic condensation of a feed stream in presence of a lipase followed by a fractionation in short path evaporator and a further fractionation in short path evaporator. The feed stream may be a condensate stream from a deodorization step.

Description

PT-2063-WO-PCTPROCESS FOR COLLECTING MINOR COMPONENTS WITH DOUBLE PASS AND USING ENZYMECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of European Application No.24213745.3, filed November 18, 2024, which is incorporated by reference herein in its entirety.FIELD OF THE INVENTION

[0002] The present invention relates to a process for obtaining minor components from vegetable oils. The minor components of interest are squalene, tocopherols, and optionally tocotri enols.BACKGROUND OF THE INVENTION

[0003] Squalene is a widely distributed natural triterpene, and it is a minor component of high commercial value worldwide because it has nutritional, pharmaceutical, medicinal, and cosmetic applications, due to its different biological properties. The main source of extraction has been shark liver oil. Secondary sources are mainly vegetable oils, although a limited one, as they allow low productive yields. Due to the diversity of applications that squalene presents and its growing demand, there is an increasing interest in identifying sustainable sources of extraction and obtaining a certain concentration / purity.

[0004] Tocopherols and tocotrienols are both part of the vitamin E family and a have a wide range of beneficial applications. The most common application is the use for their anti-oxidative properties.

[0005] Squalene, tocopherols and / or tocotrienols are present in a variety of vegetable oils. Isolation of minor components from various vegetable oils and by-products of refining of vegetable oils is less efficient as minor components are present in low concentrations. There are several processes available to concentrate and purify these minor components but they involve solvents and / or chemicals, or chromatographic separation methods.

[0006] There is a need for a suitable process to isolate these minor components in higher purify and / or yield, higher quality; for a simple process, preferably without involving solventsPT-2063-WO-PCTand / or chemicals.

[0007] The present invention provides for such a process.SUMMARY OF THE INVENTION

[0008] The present invention relates to a process for obtaining minor components from vegetable oils, the process is comprising:a) Enzy matic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;b) Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of 140°C to 180°C and a pressure below 1 mbar;c) Collecting the distillate and / or the residue; andd) further fractionating the residue of step c) into a second distillate and a second residue in a short path evaporator at a temperature of from 180°C to 240°C, below 1 mbar; Wherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; andWherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 5.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 4.0 wt.% of the feed stream.DETAILED DESCRIPTION

[0009] The present invention relates to a process for obtaining minor components from vegetable oils, the process is comprising:a) Enzymatic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;a. Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of 140°C to 180°C and a pressure below 1 mbar;b. Collecting the distillate and / or the residue; andb) further fractionating the residue of step c) into a second distillate and a second residue in a short path evaporator at a temperature of from 180°C to 240°C, below 1 mbar;PT-2063-WO-PCTWherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; andWherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols, are present in the feed stream in an amount of 0.1 to 5.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 4.0 wt.% of the feed stream.Vegetable Oils

[0010] The primary components of most unrefined vegetable oils are comprising triglycerides (TAG), diglycerides (DAG), monoglycerides (MAG), free fatty acids (FFA), minor components and the like.

[0011] Vegetable oils comprises one or more edible fats or oils chosen among palm oil, rapeseed oil and its varieties, soybean oil, sunflower oil and its varieties, shea butter, peanut oil, hazelnut oil, rice bran oil, corn oil, safflower oil. linseed oil, hempseed oil, grapeseed oil, sea buckthorn oil, walnut oil, olive oil, kapoc oil, sesame oil, almond oil, avocado oil etc. evening primrose oil, shea butter, sal fat, enzymatically prepared fats, allanblackia seed oil, illipe butter, mango fat, kokum fat, cacao butter, coconut oil, palm and palm kernel oil. The oils and fats may include single or blended oils and fats, or processed oils and fats such as fractionated.

[0012] In an aspect of the invention, the vegetable oils include palm oil or palm oil fractions, sunflower oil, rapeseed oil, soybean oil, rice bran oil, palm kernel oil, coconut oil, shea butter or mixtures thereof.

[0013] In a preferred aspect of the invention, the vegetable oil includes palm oil, palm olein, palm mid-fraction or mixtures thereof.

[0014] Unrefined vegetable oils often contain varying amounts of other components (beyond the triglycerides, diglycerides, monoglycerides), too. Some of these components are desirable components of the oil, e g., squalene, tocopherols, and / or tocotrienols, phytosterols (free and esterified), and the like.

[0015] Vegetable oil impurities (including free fatty acids) are typically removed in the refining process, in different steps, particularly degumming, alkali treatment, bleaching, and / or deodorization.PT-2063-WO-PCT

[0016] In the deodorization step, remaining volatile impurities are removed to yield a deodorized vegetable oil having the desired final characteristics. The volatile impurities removed in the deodorization process commonly include free fatty acids (FFA), aldehydes, ketones, alcohols, and other hydrocarbon impurities.

[0017] Deodorization is typically carried out at elevated temperatures and reduced pressure to better volatilize the FFAs and other impurities. The precise temperature and pressure may vary depending on the nature and quality of the oil being processed. The pressure, for instance, will preferably be no greater than 10 mbar but in certain occasions may benefit from a pressure below or equal to 5 mbar. e.g. 1 to 3 mbar. The temperature in the deodorizer may be varied as desired to optimize the yield and quality of the deodorized oil. At higher temperatures, reactions which may degrade the quality' of the oil will proceed more quickly. For example, at higher temperatures, cis-fatty acids may be converted into their less desirable transform. Operating the deodorizer at lower temperatures may minimize the cis-to-trans conversion but will generally take longer or require more stripping medium or lower pressure to remove the requisite percentage of volatile impurities. As such, deodorization is typically performed at a temperature of the oil in a range of 180 to 270°C, with temperatures of about 220-260°C being useful for many oils. For cocoa butter-based oil, a deodorization temperature in a range of 130 to 220°C is advised. Typically, deodorization is thus occurring in a deodorizer whereby volatile components such as FFAs and other unwanted volatile components that may cause off-flavors in the oil, are removed. Deodorization may also result in the thermal degradation of components.

[0018] Unfortunately, some of the desirable components such as the minor components of the vegetable oil may be driven off with the volatile impurities during the deodorization process. As a consequence, the volatile stream exiting the deodorizer will include a substantial fraction but may also include a varying amount of desirable components.Minor Components

[0019] In an aspect of the invention, the minor components are tocopherols, tocotrienols, triterpenes (such as squalene, alfa-amyrin, beta-amyrin and the like), phytosterols (free sterols and sterol esters).

[0020] In an aspect of the invention, the minor components are squalene, tocopherols, tocotrienols and phytosterols.PT-2063-WO-PCT

[0021] In a further aspect of the invention, the minor components are squalene, tocopherols and tocotrienols.

[0022] In another aspect of the invention, the minor components are squalene and tocopherols.Feed Stream

[0023] Feed stream is the starting material used for the enzymatic condensation step in presence of a lipase.

[0024] The feed stream is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene, and optionally tocotrienols and; and wherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 5.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 4.0 wt.% of the feed stream.

[0025] In an aspect of the invention, the feed stream is comprising tocopherols in an amount of 0.1 to 3.0 wt.% of the feed stream.

[0026] In an aspect of the invention, the feed stream is comprising squalene in an amount of 0.1 to 2.0 wt.% of the feed stream.

[0027] In an aspect of the invention, the feed stream is comprising phytosterols in an amount of 0.7 to 6.0 wt.% of the feed stream.

[0028] In an aspect of the invention, the feed stream is comprising triglycerides in an amount of 7 to 55 wt.% based on weight of the feed stream.

[0029] In an aspect of the invention, the feed stream is comprising diglycerides in an amount of 15 to 35 wt.% based on weight of the feed stream.

[0030] In an aspect of the invention, the feed stream is comprising monoglycerides in an amount of 1 to 4.5 wt.% based on weight of the feed stream.

[0031] In an aspect of the invention, the feed stream is comprising tocotrienols in an amount of 0.1 to 2 wt.% of the feed stream.

[0032] In an aspect of the invention, the feed stream is sourced from vegetable oils that include palm oil or palm oil fractions, sunflower oil, rapeseed oil, soybean oil, rice bran oil, palm kernel oil. coconut oil, shea butter or mixtures thereof.

[0033] In an aspect of the invention, the feed stream is sourced from vegetable oil including palm oil, palm olein, palm mid-fraction or mixtures thereof.PT-2063-WO-PCT

[0034] In an aspect of the invention, the feed stream is a condensate stream of a deodorization step of a vegetable oil.

[0035] A deodorization step of a vegetable oil can be conducted such that a deodorized vegetable oil and a volatiles stream is obtained. A condensation stream can be obtained by partial and / or complete condensation of the volatiles stream. The condensation stream is cooled at a certain temperature, here referred to as the condenser temperature. The condensation stream comprises a portion of the glycerides and a majority of the volatile impurity fraction of the vegetable oil, as well as the minor components of interest. Such a condensate stream may be the feed stream of the present invention.

[0036] In an aspect of the invention, the condensate stream is obtained at a condenser temperature of from 100°C to 180°C, preferably from 140°C to 160°C.

[0037] In an aspect of the invention, the feed stream, preferably a condensate stream (condensate from the deodorization distillate) is comprising:- triglycerides in an amount of 7 to 55 wt.%, preferably from 20 to 40 wt.% based on weight of the feed stream,diglycerides in an amount of 15 to 35 wt. %, preferably from 20 to 32 wt. % based on weight of the feed stream,- monoglycerides in an amount of 1 to 4.5 wt.% preferably from 2.0 to 4.0 wt.% based on weight of the feed stream,- free fatty acids in an amount of 5 to 65 wt.% based on weight of the feed stream; preferably from 15 to 55 wt.% based on weight of the feed stream, preferably from 20 to 35 wt.% based on weight of the feed stream,- tocopherols in an amount of 0.1 to 5.0 wt.% based on weight of the feed stream, preferably from 0.1 to 3.0 wt.% based on weight of the feed stream,squalene in an amount of 0.1 to 4.0 wt.% based on weight of the feed stream, preferably from 0.1 to 2.0 wt.% based on weight of the feed stream; and- phytosterols in an amount of 0.7 to 6.0 wt.% of the feed stream.

[0038] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof, and the feed stream, preferably a condensate stream is comprising:PT-2063-WO-PCT- triglycerides in an amount of 7 to 55 wt.%, preferably from 20 to 40 wt.% based on weight of the feed stream,diglycerides in an amount of 15 to 35 wt. %, preferably from 20 to 32 wt. % based on weight of the feed stream,- monoglycerides in an amount of 1 to 4.5 wt.% preferably from 2.4 to 4.0 wt.% based on weight of the feed stream,free fatty acids in an amount of 5 to 65 wt.% based on weight of the feed stream; preferably from 15 to 35 wt.% based on weight of the feed stream,- tocopherols and tocotrienols in an amount of 0.1 to 3.0 wt.% based on weight of the feed stream;squalene in an amount of 0.1 to 2.0 wt.% based on weight of the feed stream; and phytosterols in an amount of 0.7 to 6.0 wt.% of the feed stream.Enzymatic Condensation

[0039] The feed stream is enzymatically treated (= enzymatic condensation) in presence of a lipase.

[0040] Optionally, the enzymatic condensation of the feed stream is taking place in presence of a lipase and glycerol.

[0041] In a preferred aspect of the invention, the feed stream is containing sufficient hydroxyl groups for the condensation of the fatty acids, and there is no need to add glycerol in step a) (enzymatic condensation).

[0042] The lipase may be used in solution or can be immobilized and as such have an impact on the purification of the reaction medium and stability of the enzyme.

[0043] Lipases are classified according to the sources from which they are obtained, such as microorganism (fungi or bacteria), animal and plant. Most lipases applied are derived from fungal or bacterial sources. Some of the most widely used fungal lipases are derived from various species within genera such as Candida, Yarrowia, Aspergillus and Penicillium, while bacterial lipases often come from Pseudomonas sp., Bacillus sp., Staphylococcus sp., Burkholderia sp. and many others. Bacterial lipases and esterases have been classified into eight families (and several subfamilies) based on sequence homology and biological properties. Lipases from animal origin have been originated from various organs and tissues of several mammalian species, among which the pancreatic lipases are the most thoroughly studied. Plant lipases have not received the samePT-2063-WO-PCTattention as those from other sources, but oilseed lipases have been of greatest interest among the plant lipases. An non-exhaustive, non-limiting list of suitable lipases includes, lipase derived from Candida cylindracea, Candida lipolytica, Candida rugosa, Candida antarctica, Candida utilis, Chromobacterium viscosum, Geotrichum viscosum, Geotrichum candidum, Mucor javanicus, Mucor miehei, Porcine pancreas. Pseudomonas species, specifically Pseudomonas fluorescens. Pseudomonas cepacia, Pseudomonas pseudoalkaligenes, Pseudomonas alkaligenes, Thermomyces species, Rhizopus arrhizus, Rhizopus delemar, Rhizopus niveus, Rhizopus oryzae. Rhizopus javanicus, Aspergillus niger, Penicillium roquefortii, Penicillium camembertii or an esterase derived from Bacillus species, specifically Bacillus thermoglucosidasius; Mucor miehei, Horse liver, Saccharomyces cerevisiae, Pigs liver or combinations thereof.

[0044] In an aspect of the invention, the lipase is added in an amount of 1 to 3 wt.% based upon the weight of the feed stream.

[0045] The enzymatic treatment is conducted at a temperature of 60 to 75°C. preferably at a temperature of 68 to 72°C.

[0046] The product obtained after the enzymatic treatment is the enzymatic treated feed stream.In an aspect of the invention, the enzymatic treated feed stream is comprising:i. Triglycerides in an amount of 50 to 90 wt.% based on weight of the enzymatic treated feed stream;ii. Free fatty acids in an amount of 1 to 10 wt.% based on weight of the enzymatic treated feed stream, preferably from 1 to 5 wt.% based on weight of the enzymatic treated feed stream;iii. Tocopherols in an amount of 0.1 to 5.0 wt.% based on weight of the enzymatic treated feed stream; andiv. Squalene in an amount of 0.1 to 4.0 wt.% based on weight of the enzymatic treated feed stream.

[0047] In an aspect of the invention, the enzymatic treated feed stream is comprising:i. Triglycerides in an amount of 50 to 90 wt.% based on weight of the enzymatic treated feed stream;PT-2063-WO-PCTii. Free fatty acids in an amount of 1 to 10 wt.% based on weight of the enzymatic treated feed stream, preferably from 1 to 5 wt.% based on weight of the enzymatic treated feed stream;iii. Tocopherols in an amount of 0.1 to 3.0 wt.% based on weight of the enzymatic treated feed stream; andiv. Squalene in an amount of 0.1 to 2.0 wt.% based on weight of the enzymatic treated feed stream.

[0048] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the enzymatic treated feed stream is comprising:i. Triglycerides in an amount of 50 to 90 wt.% based on weight of the enzymatic treated feed stream;ii. Free fatty’ acids in an amount of 1 to 10 wt.% based on weight of the enzymatic treated feed stream preferably from 1 to 5 wt.% based on weight of the enzymatic treated feed stream;iii. Tocopherols and tocotrienols in an amount of 0.1 to 3.0 wt.% based on weight of the enzymatic treated feed stream; andiv. Squalene in an amount of 0.1 to 2.0 wt.% based on weight of the enzymatic treated feed stream.

[0049] In an aspect of the invention, the enzymatic treated feed stream is comprising phytosterols in an amount of 0.7 to 6.0 wt.% of the feed stream.

[0050] In an aspect of the invention, it has been shown that the enzymatic treated feed stream has a reduced content of free fatty acids of at least 50%, preferably at least 70%, more preferably at least 80%.Fractionation

[0051] The enzymatic treated feed stream is fractionated in short path evaporator (SPE).

[0052] The fractionation of step b) is conducted in a short path evaporation at a temperature of 140°C to 180°C.

[0053] Short-path evaporation, also called short-path distillation or molecular distillation, is a distillation technique that involves the distillate travelling a short distance, often only a fewPT-2063-WO-PCTcentimetres, and it is normally done at reduced pressure. With short path distillation, a decrease of boiling temperature is obtained by reducing the operating pressure. It is a continuous process with very short residence time. This technique is often used for compounds which are unstable at high temperatures or to purify small amounts of compounds. The advantage is that the heating temperature can be considerably lower (at reduced pressure) than the boiling point of the liquid at standard pressure. Additionally, short-path evaporation allows working at very low pressure.

[0054] Different types of short-path evaporation apparatus can be used that are well known to the skilled person. Examples are. but are not limited to, falling film, centrifugal, or wiped film evaporation apparatus. Preferably the short-path evaporation of the current process is performed in a wiped film evaporation apparatus.

[0055] The short-path evaporation in step b) of the process according to the invention is performed at a pressure below 1 mbar, preferably below 0.1 mbar, more preferably below 0.01 mbar, most preferably below 0.005 mbar.

[0056] The short-path evaporation is further performed at specific conditions of temperature and feed rate per unit area of evaporator surface of the shorth-path evaporation equipment.

[0057] The ‘‘feed rate per unit area of evaporator surface of the shorth-path evaporation equipment", also called “specific throughput’' or “specific feed rate”, expressed in kg / h per m2, is defined as the flow of oil, expressed in kg / h, per unit area of evaporator surface of the short-path evaporation equipment, expressed in m2The feed rate per unit area of evaporator surface of the shorth-path evaporation equipment in the process of the current invention is applicable to any short-path equipment, including industrial short-path evaporation equipment independent of the dimensions of the equipment. Preferably stainless steel short-path evaporation equipment is used in the current invention.

[0058] In an aspect of the invention, the short path evaporator is operated with a feed rate (flow rate) (kilogram per hour) per unit area of evaporator surface of the shorth-path evaporation equipment in a range of from 10 to 200 kilogram per hour per square meter (kg / h per m2), preferably from 12 to 150 kilogram per hour per square meter (kg / h per m2), more preferably in a range of from 15 to 100 kilogram per hour per square meter (kg / h per m2), even more preferably in a range of from 17 to 80 kilogram per hour per square meter (kg / h per m2), most preferably in a range of from 20 to 50 kilogram per hour per square meter (kg / h per m2).PT-2063-WO-PCT

[0059] The fractionation of the enzymatic treated feed stream provides a distillate and a residue.

[0060] In an aspect of the invention, the distillate collected in step c) is comprising i. Tocopherols in an amount of 0.1 to 7 wt.% based on weight of the distillate; andii. Squalene in an amount of 2 to 15 wt.% based on weight of the distillate.

[0061] In an aspect of the invention, the distillate collected in step c) is further comprising i. Triglycerides in an amount of less than 5 wt.% based upon the weight of the distillate;ii. Free fatty acids in an amount of 70 to 97 wt.% based on weight of the distillate.

[0062] In an aspect of the invention, the distillate collected in step c) is comprising i. Triglycerides in an amount of less than 5 wt.% based upon the weight of the distillate;ii. Diglycerides in an amount of 0.1 to 2 wt.% wt.% based on weight of the distillate:iii. Monoglycerides in an amount of 0.1 to 4 wt.% wt.% based on weight of the distillate;iv. Free fatty acids in an amount of 70 to 97 wt.% based on weight of the distillate:v. Tocopherols in an amount of 0.1 to 7 wt.% based on weight of the distillate;andvi. Squalene in an amount of 2 to 15 wt.% based on w eight of the distillate;

[0063] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the distillate collected in step c) is comprising:iii. Tocopherols and tocotrienols in an amount of 0.1 to 7 wt.% based on weight of the distillate;iv. Squalene in an amount of 2 to 15 wt.% based on weight of the distillate;PT-2063-WO-PCT

[0064] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the distillate collected in step c) is further comprising:i. Triglycerides in an amount of less than 5 wt.% based upon the weight of the distillate;ii. Free fatty acids in an amount of 70 to 97 wt.% based on weight of the distillate.

[0065] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the distillate collected in step c) is comprising:i. Triglycerides in an amount of less than 5 wt.% based upon the weight of the distillate;ii. Diglycerides in an amount of 0.1 to 2 wt.% wt.% based on weight of the distillate:iii. Monoglycerides in an amount of 0.1 to 4 wt.% wt.% based on weight of the distillate;iv. Free fatty acids in an amount of 70 to 97 wt.% based on weight of the distillate:v. Tocopherols and tocotrienols in an amount of 0.1 to 7 wt.% based on w eight of the distillate;vi. Squalene in an amount of 2 to 15 wt.% based on w eight of the distillate.

[0066] In an aspect of the invention, the distillate is obtained in an amount of 1.5 to 8 wt.% of the enzymatic treated feed stream (mass balance).

[0067] In the fractionation of step b) beyond the distillate that can be collected, also a residue is obtained.In an aspect of the invention, the residue collected in step c) is comprisingi. Tocopherols in an amount of 0.3 to 4.0 wt.% based on weight of the residue; ii. Squalene in an amount of 0.3 to 3.0 wt.% based on weight of the residue.

[0068] In an aspect of the invention, the residue collected in step c) is comprising phytosterols in an amount of 3 to 8 wt.% based on weight of the residue.PT-2063-WO-PCT

[0069] In an aspect of the invention, the residue collected in step c) is further comprising i. Triglycerides in an amount of 55 to 75 wt.% based upon the weight of the residue;ii. Diglycerides in an amount of 5 to 15 wt.% based on weight of the residue.

[0070] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the residue collected in step c) is comprising i. Tocopherols and tocotrienols in an amount of 0.5 to 4.0 wt.% based on weight of the residue;ii. Squalene in an amount of 0.3 to 3.0 wt.% based on weight of the residue.

[0071] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the residue collected in step c) is further comprisingiii. Triglycerides in an amount of 55 to 75 wt.% based upon the weight of the residue;iv. Diglycerides in an amount of 5 to 15 wt.% based on weight of the residue.

[0072] In an aspect of the invention, the residue is obtained in an amount of 92 to 98.5 wt.% of the enzymatic treated feed stream (mass balance).

[0073] In the process of the present invention, the residue is further fractionated by applying a short path evaporator at a temperature of from 180°C to 240°C, for obtaining second distillate and a second residue.

[0074] In an aspect of the invention, the further fractionation is conducted in a short path evaporator, at a temperature of from 185°C to 235°C, preferably at a temperature of from 190°C to 230°C, more preferably at a temperature of from 200°C to 220°C or from 210°C to 215°C.

[0075] In the present invention, the fractionation of step b) is conducted in a short path evaporator at a temperature of 140°C to 180°C, followed by a further fractionation in a short path evaporator at a temperature of from 180°C to 240°C, preferably at a temperature of from 185°C to 235°C, 190°C to 230°C. more preferably at a temperature of from 200°C to 220°C or from 210°C to 215°C.PT-2063-WO-PCT

[0076] The short-path evaporation (i.e. fractionation and further fractionation) is performed at a pressure below 1 mbar, preferably below 0.1 mbar, more preferably below 0.01 mbar, most preferably below 0.005 mbar.

[0077] In an aspect of the invention, the short path evaporator is operated with a feed rate (flow rate) (kilogram per hour) per unit area of evaporator surface of the shorth-path evaporation equipment in a range of from 10 to 200 kilogram per hour per square meter (kg / h per m2),

[0078] The further fractionation is either applying a second short path equipment or alternatively it involves a second pass on the same equipment wherein the process conditions of the further fractionation are applied.

[0079] In an aspect of the invention, the second distillate is comprising:i. Tocopherols in an amount of 20 to 45 wt.% based on weight of the distillate; andii. Squalene in an amount of 7 to 15 wt.% based on weight of the distillate.

[0080] In an aspect of the invention, the second distillate is comprising phytosterols in an amount of 9 to 18 wt.% based on weight of the distillate.

[0081] In an aspect of the invention, the second distillate is comprising:i. Triglycerides in an amount of less than 1 wt.% based upon the weight of the distillate;ii. Diglycerides in an amount of 5 to 25 wt.% wt.% based on weight of the distillate;iii. Monoglycerides in an amount of 1.5 to 9 wt.% wt.% based on weight of the distillate;iv. Free fatty acids in an amount of 10 to 25 wt.% based on weight of the distillate.

[0082] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the second distillate is comprising:iii. Tocopherols and tocotrienols in an amount of 20 to 45 wt.% based on weight of the distillate; andiv. Squalene in an amount of 7 to 15 wt.% based on weight of the distillate.PT-2063-WO-PCT

[0083] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the second distillate is comprising:i. Triglycerides in an amount of less than 1 wt.% based upon the weight of the distillate;ii. Diglycerides in an amount of 5 to 25 wt.% wt.% based on weight of the distillate;iii. Monoglycerides in an amount of 1.5 to 9 wt.% wt.% based on weight of the distillate:iv. Free fatty acids in an amount of 10 to 25 wt.% based on weight of the distillate.

[0084] In an aspect of the invention, the vegetable oils are selected from palm oil, palm olein, palm mid-fraction or mixtures thereof and the second distillate is comprising phytosterols in an amount of 9 to 18 wt.% based on weight of the distillate.

[0085] In an aspect of the invention, the second distillate is obtained in an amount of 3 to 15 wt.%, preferably 3.5 to 10 wt.% of the weight of the residue (is mass balance). This is the residue from first fractionation at 140-180°C.

[0086] The residue of the first fractionation using SPE is further fractionated and the obtained second residue is collected.

[0087] In an aspect of the invention, it relates to a process for obtaining minor components from vegetable oils, and the process is comprising:a) Enzymatic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;b) Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of from 140 up to 180°C:c) Further fractionation of the residue of step b) into a second distillate and a second residue and the fractionation is taking place in a short path evaporator at a temperature of from 180°C to 240°C;d) Collecting the distillate and / or collecting the second distillate;e) Collecting the residue and / or second residue; andPT-2063-WO-PCTWherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; andWherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 5.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 4.0 wt.% of the feed stream.

[0088] In an aspect of the invention, it relates to a process for obtaining minor components from vegetable oils, and the process is comprising:i. Enzymatic condensation of a feed stream and glycerol in presence of a lipase for obtaining an enzymatic treated feed stream;ii. Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of from 140 up to 180°C;iii. Further fractionation of the residue of step b) into a second distillate and a second residue and the fractionation is taking place in a short path evaporator at a temperature of from 180°C to 240°C;iv. Collecting the distillate and / or collecting the second distillate;v. Collecting the residue and / or second residue; andWherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; andWherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 3.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 2.0 wt.% of the feed stream.

[0089] In an aspect of the invention, it relates to a process for obtaining minor components from vegetable oils selected from palm oil, palm olein, palm mid-fraction or mixtures thereof, and the process is comprising:a) Enzymatic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;PT-2063-WO-PCTb) Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of from 140 up to 180°C;c) Further fractionation of the residue of step b) into a second distillate and a second residue and the fractionation is taking place in a short path evaporator at a temperature of from 180°C to 240°C;d) Collecting the distillate and / or collecting the second distillate;e) Collecting the residue and / or second residue; andWherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, squalene, tocopherols and tocotrienols; and Wherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols and tocotrienols are present in the feed stream in an amount of 0.1 to 3.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 2.0 wt.% of the feed stream.

[0090] The advantages of the present invention are amongst others:- Isolation of minor components from various streams is difficult as they are present in low concentrations; but the process of the present invention allow s the concentration of the minor components without using solvents and / or chromatographic separation methods; - Upgrade the value of existing condensation stream obtained in deodorization process of edible oils;- The enzymatic treatment allowed to obtain a stream with a significant reduction of the content of free fatty acids, while the amount of minor components remained (quasi) unchanged;- There is no need to add glycerol in the enzymatic condensation step;- Avoiding long heat treatments and thus reduce breakdown of the heat-sensitive minor components;- Provision of distillates with high value due to the high amount of minor components, that can have multiple applications;- Pre-enriches minor components, which helps further process capabilities to operate at higher throughput / higher capacity;PT-2063-WO-PCT- Use of short path evaporator (double pass) allows a smooth / mild process;- The residue and / or second residue can be re-cycled back into the overall process of deodorization of vegetable oil.

[0091] The present invention is illustrated by the following examples.EXAMPLESAnalytical Methods

[0092] Lipid Profile by GC / FID

[0093] Lipid Profile analysis was used to determine the lipid composition of the samples ((monoglycerides, diglycerides, triglycerides, free fatty acids, glycerol, phytosterols (free and esterified sterols, including ferulates), squalene, hydrocarbons)), within a single analysis. Quantitation of each class of compounds used multi-level calibration curves with an appropriate standard. The sample size was approximately 100 mg. An internal standard (IS) of heptadecanyl stearate (HDS) was used at 10 mg. Samples were silylated with N, 0,-bis-(trimethylsilyl) trifluoroacetoamide (BSTFA) with 1% trimethylchlorosilane and pyridine. The trimethylsilane (TMS) ethers were analyzed by cool on-column (COC) gas chromatography (GC) with anon-polar column stationary phase (15 mx 0.25 mmx 0.10 mm df, DB™-5HT) coupled to a flame ionization detector (FID). The temperature program was 110°C (0.2 min) to 140°C at 30°C / min to 340°C at 10°C / min (13.8 min). Hydrogen was the carrier gas, and inlet pressure was 6.7 psi at 110°C in the constant flow mode. The detector temperature was 370°C. The FID air flow rate was 450 mL / min, the FID hydrogen flow rate was 40 mL / min, and the makeup gas flow was 40 mL / min. Samples were analyzed a single time.

[0094] Analysis of Total and Isomeric Tocopherols / Tocotrienols by Liquid Chromatography with Fluorescence Detection (LC / FLD). Total and isomeric tocopherol / tocotrienol concentrations in the samples were determined by LC / FLD. A five-point calibration curve was created using an established tocopherol mix that contained a-Tocopherol, b-Tocopherol. g-Tocopherol, and d-Tocopherol; the tocotrienol isomers referenced their corresponding tocopherol calibration curve. Oil samples were prepared at 200 mg / mL (2 g in a 10 mL volumetric and diluted to volume with n-hexane) and distillate preparations ranged from 1 to 100 mg / mL oil depending on tocopherol and tocotrienol concentration. Chromatography wasPT-2063-WO-PCTperformed on a 250 mm x 4.6 mm LiChrospher® 60 (5mm particle size) HPLC column using a mobile phase of 99% n-Hexane with 1% Isopropyl Alcohol running under isocratic conditions (flow rate at 1.0 mL / min) with a 10 mL injection volume. The fluorescence detector was set to run an excitation wavelength of 290 nm and collect the emission at a wavelength of 330 nm. The calibration curves were linear through zero.Example 1The feed stream (condensate stream)

[0095] The condensate stream obtained from palm oil common refining process was used as a feed stream. The refining process of the palm oil included a deodorization performed at 265°C.

[0096] The feed stream can be obtained at different condenser temperature settings. The corresponding compositions can be seen below in Table 1 and Table 2.Table 1: Minor Components in Feed StreamTotalTotal TotalCondenser Tocopherol Total Tocopherol Tocotrienol Squalenetemperature s and phytosterolss s [wt.%]settings (°C) tocotrienols [wt %][wt %] [wt %][wt.%]120 0.30 0.84 1.14 1.56 3.25130 0.16 0.73 0.89 1.16 4.32140 0.27 0.72 0.99 1.02 5.16150 0.46 1.76 2.22 1.34 2.68160 0.37 1.26 1.63 0.69 1.47170 0.27 0.92 1.19 0.49 1.40180 0.16 0.61 0.77 0.35 1.38Table 2: Glycerides Composition of Feed StreamCondenserTAG DAG MAG FFAtemperature[wt.%] [wt.%] [wt.%] [wt.%]settings (°C)120 12.12 18.96 2.71 54.95130 20.99 21.48 2.68 42.44140 24.51 24.97 2.69 33.31150 25.01 28.81 3.76 20.72PT-2063-WO-PCT160 38.97 29.61 2.32 16.04170 44.46 30.23 1.94 10.68180 48.87 30.85 1.55 7.15Enzymatic condensation of the feed stream

[0097] The feed stream had a starting FFA content of 20.7% (see table 2, condensation at 150°C).

[0098] The enzyme Lipozyme 435 was used for the enzymatic condensation reaction. The feed stream (36.2 kg) was heated to 70°C under vacuum (1-2 mbar) to reduce moisture. The dosage of 2% Lipozyme 435 was added to the feed stream and allowed to react for 20 and 22 hours, respectively. The free fatty acids content was reduced to 4 wt.%.

[0099] The enzymatic treatment had relatively no impact on the amount of squalene, tocopherols, and / or tocotrienols.

[0100] The corresponding composition can be seen below in Table 3 and Table 4.Table 3: Minor components in Enzymatic Treated Feed StreamTotalTotal Total Total Tocopherols SqualeneTocopherols tocotrienols phytosterols +tocotrienols [wt %][wt %] [wt %] '[wt %][wt %]Enzymatic treated0.53 1.96 2.51 1.57 5.45feed streamTable 4: Glycerides Composition of Enzy matic Treated Feed StreamTotal TotalTotal TAG Total MAGDAG [wt FFA [wt[wt %] [wt %]%] %]Enzymatictreated feed 63.10 8.71 0.20 4.08streamFractionation - Short Path Evaporation (SPE) of the enzymatic treated feed stream (1ststep)

[0101] Short-Path Evaporation (SPE) Unit KDL-5 from UIC was used for the short path evaporation tests. The KDL-5 unit has an evaporator surface of 0.048 m2. The enzymatic treated feed stream was fractionated on the SPE to obtain a distillate and residue.For the tests, the pressure and flow rate were kept relatively constant (see process conditions inPT-2063-WO-PCTTable 5) while evaporator temperature was changed (example to 140°C, 160°C or 180°C).Table 5: Process settings for SPE testsTest-conditions Unit Test 140°C Test 160°C Test 180°C Temperatures bathfeed (°C) 80 80 80 evaporator (°C) 140 160 180residue (°C) 70 70 80condenser (°C) 70 70 70Flow-rate perevaporator (kg / h per m2) 29 29 29surface areaVacuum systempressure (mbar) 2.0*10-3 2.1*10-3 2.0*10-3 Wiper Systemwiper type rollers / blocks rollers rollers rollersrotation speed (rpm) 366 366 366

[0102] The mass balances and compositions of the obtained distillates and residues, including content of free fatty acids, tocopherol, tocotrienols, phytosterols, and squalene, after SPE treatment at different temperatures, can be seen in Table 6, 7 & 8.Table 6: Mass balance 1stfractionation SPEMass balance,wt.%SPE@ 140°CDistillate 2.30Residue 97.60SPE@ 160°CDistillate 3.90Residue 96.10SPE@180°CDistillate 5.40Residue 94.60Table 7: Minor Components of the distillates and residues 1stfractionation SPEPT-2063-WO-PCTTotalTotal Total Tocopherols Total Squalene Tocopherols tocotrienols and phytosterols [wt %][wt %] [wt %] Tocotrienols [wt %][wt %]SPE@ 140°CDistillate ND 0.10 0.10 3.33 0.21 Residue 0.55 2.02 2.57 1.22 5.63SPE@ 160°CDistillate 0.39 0.63 1.02 7.57 0.73 Residue ND 1.87 1.87 1.00 5.65SPE@180°CDistillate 1.38 4.09 5.49 12.56 2.00 Residue ND 0.78 0.78 0.67 5.82ND=not detectedTable 8: Glyceride Compositions of the distillates and residues 1stfractionation SPETotal TAG Total DAG Total MAG Total FFA[wt %] [wt %] [wt %] [wt %]SPE@ 140°CDistillate ND 0.56 0.59 95.21Residue 64.82 9.19 0.29 1.89SPE@ 160°CDistillate ND 0.10 1.41 85.95Residue 66.64 8.20 0.69 1.43SPE@180°CDistillate ND 0.61 2.19 75.20Residue 67.21 9.48 0.22 0.38ND=not detectedFurther Fractionation: Short Path Evaporation (SPE) of the Residue2-Step SPE process

[0103] The residue from the first SPE (fractionation) was collected and furtherPT-2063-WO-PCTfractionated on an SPE KDL-5 equipment, and the process conditions are shown in Table 9.Table 9: Process settings for SPE - further fractionationTest-condition Unit Test 220°C Test 240°CTemperatures bathfeed (°C) 80 80evaporator (°C) 220 240residue (°C) 120 140condenser (°C) 70 70Flow-rate per evaporation(kg / h per m2) 29 29surface areaVacuum systempressure (mbar) 2.1*10-3 2.0*10-3Wiper Systemwiper type rollers / blocks rollers rollersrotation speed (rpm) 366 366

[0104] The mass balance and compositions of the second distillate and second residue can be seen in Table 10, 11 & 12.Table 10: Mass balance further fractionation SPEMass balance,[wt. %]SPE@ 160°C=> SPE@ 220°CDistillate 5.50Residue 90.60SPE@ 160°C=> SPE@ 240°CDistillate 7.80Residue 88.20SPEz / . 180°C=> SPE@ 220°CDistillate 4.40Residue 90.20SPE@ 180°C=> SPE@ 240°CDistillate 6.90Residue 87.70PT-2063-WO-PCTTable 11: Minor Components of the second distillate and second residue after further fractionation SPETotalTotal Total Total Tocopherols Squalene Tocopherols tocotrienols phytosterols +tocotrienols [wt %][wt %] [wt %] [wt %][wt %]Residue0.53 1.34 1.87 1.00 5. 65 1stSPE 160°CSPJ 160°C=> SPE(®y220°CDistillate 6.50 22.70 29.20 12.11 11.36 Residue 0.15 0.59 0.73 0.11 5.31 SPE@ 160°C=> SPE@ 240°CDistillate 6.01 20.70 26.71 10.77 11.52Residue 0.07 0.24 0.32 0.06 4.92 Table 12: Glycerides Compositions of the second distillate and second residue after further fraction SPETotal TAG Total DAG Total MAG Total FFA[wt %] [wt %] [wt %] [wt %]Residue66.64 8.20 0.69 1.431stSPE 160°CSPE? / , 160°C=> SPE? / 220°CDistillate 10.88 6.84 19.18Residue 67.66 9.06SPE? / , 160°C=> SPE? / , 2ZIO°CDistillate 0.49 19.40 2.53 12.45Residue 71.27 7.84 ND NDdetectedTable 13: Minor Components of the second distillate and second residue after further fractionation SPEPT-2063-WO-PCTTotalTotal Total Total Tocopherols Squalene Tocopherols tocotrienols phytosterols +tocotrienols [wt %][wt %] [wt %] [wt %][wt %]Residue 0.48 0.30 0.78 0.67 5.82 1stSPE 180°CSPEU 180°C=> SPEA 220°CDistillate 7.71 27.54 35.31 10.25 15.36 Residue 0.13 0.51 0.64 0.04 4.82SPEA, 180°C=> SPEA. 240°CDistillate 6.68 24.23 30.91 9.42 13.4Residue 0.06 0.25 0.32 0.08 5.29 Table 14: Glycerides Compositions of the second distillate and second residue after further fractionation SPETotalTotal TAG Total DAG Total FFAMAG [wt[wt %] [wt %] [wt %]%1Residue67.21 9.48 0.22 0.381stSPE 180°CSPEA, 180°C=> SPIN / 220°CDistillate 0.84 15.53 2.31 7.94Residue 68.08 8.73 0.02 0.02SPETz 180°C=> SPEA. 240°CDistillate 0.38 22.66 2.30 8.69Residue 71.76 7.95 ND ND ND=not detected

Claims

1. PT-2063-WO-PCT2.CLAIMS1. A process for obtaining minor components from vegetable oils, the process is comprising:4.a) Enzymatic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;5.b) Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of 140°C to 180°C and a pressure below 1 mbar;6.c) Collecting the distillate and / or the residue; and7.d) further fractionating the residue of step c) into a second distillate and a second residue in a short path evaporator at a temperature of from 180°C to 240°C, below 1 mbar;. Wherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; and8.Wherein free fatty acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 5.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 4.0 wt.% of the feed stream.

2. The process according to claim 1 wherein the feed stream is comprising:10.i. Triglycerides in an amount of 7 to 55 wt.% based on weight of the feed stream.

3. The process according to claim 1 or 2 wherein the feed stream is further comprising:12.i. Diglycerides in an amount of 15 to 35 wt. % based on weight of the feed stream; ii. Monoglycerides in an amount of 1 to 4.5 wt.% based on weight of the feed stream.

4. The process according to anyone of the preceding claims wherein the enzymatic condensation of a feed stream takes place in presence of a lipase and without glycerol.

5. The process according to anyone of the preceding claims wherein the enzymatic treated feed stream is comprising:PT-2063-WO-PCT15.i. Triglycerides in an amount of 50 to 90 wt.% based on weight of the enzymatic treated feed stream;16.ii. Free fatty acids in an amount of 1 to 10 wt.% based on weight of the enzy matic treated feed stream;17.iii. Tocopherols in an amount of 0.1 to 5.0 wt.% based on weight of the enzymatic treated feed stream; and18.iv. Squalene in an amount of 0.1 to 4.0 wt.% based on weight of the enzymatic treated feed stream.

6. The process according to anyone of the preceding claims wherein the distillate collected in step c) is comprising20.i. Tocopherols, and optionally tocotrienols in an amount of 0.1 to 7 wt.% based on weight of the distillate; and21.ii. Squalene in an amount of 2 to 15 wt.% based on weight of the distillate.

7. The process according to anyone of the preceding claims wherein the residue collected in step c) is comprising23.i. Tocopherols, and optionally tocotrienols in an amount of 0.3 to 4 wt.% based on weight of the residue; and24.ii. Squalene in an amount of 0.3 to 3 wt.% based on weight of the residue.

8. The process according to anyone of the preceding claims wherein the residue collected in step c) is comprising26.i. Triglycerides in an amount of 55 to 75 wt.% based upon the weight of the residue;27.ii. Diglycerides in an amount of 5 to 15 wt.% based on weight of the residue.

9. The process according to anyone of the preceding claims wherein the second distillate is comprising:PT-2063-WO-PCT29.i. Tocopherols, and optionally tocotrienols in an amount of 20 to 45 wt.% based on weight of the distillate; and30.ii. Squalene in an amount of 7 to 15 wt.% based on weight of the distillate10. The process according to anyone of the preceding claims wherein the feed stream is a condensate stream of a deodorization step of a vegetable oil.

11. The process according to claim 12 wherein the condensate stream is obtained at a condenser temperature of from 100°C to 180°C.

12. The process according to anyone of the preceding claims wherein the residue is collected or the second residue of anyone of claims 9 to 11 is collected.

13. The process according to anyone of the preceding claims wherein the process is comprising the following steps:35.a) Enzymatic condensation of a feed stream in presence of a lipase for obtaining an enzymatic treated feed stream;36.b) Fractionation of the enzymatic treated feed stream into a distillate and a residue and the fractionation is taking place in a short path evaporator at a temperature of from 140 up to 180°C;37.c) Further fractionation of the residue of step b) into a second distillate and a second residue and the fractionation is taking place in a short path evaporator at a temperature of from 180°C to 240°C;38.d) Collecting the distillate and / or collecting the second distillate;39.e) Collecting the residue and / or second residue; and40.Wherein the feed stream of step a) is comprising triglycerides, diglycerides, monoglycerides, free fatty acids, tocopherols, squalene and optionally tocotrienols; and41.Wherein free fatty' acids are present in the feed stream in an amount of 5 to 65 wt.%; tocopherols are present in the feed stream in an amount of 0.1 to 3.0 wt.% of the feed stream, and squalene is present in an amount of 0.1 to 2.0 wt.% of the feed stream.

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