(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/011891 Al 26 January 2017 (26.01.2017) P O P CT
(51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A23D 9/00 (2006.01) A61K 31/56 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, A23D 9/007 (2006.01) A61K 31/575 (2006.01) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, A61K 31/122 (2006.01) A61K 36/00 (2006.01) HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, A61K 31/202 (2006.01) A61P 3/02 (2006.01) KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, A61K 31/355 (2006.01) CUB 1/00 (2006.01) MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, (21) International Application Number: SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, PCT/CA20 15/000441 TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (22) International Filing Date: (84) Designated States (unless otherwise indicated, for every 23 July 20 15 (23.07.2015) kind of regional protection available): ARIPO (BW, GH, (25) Filing Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (26) Publication Language: English TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, (72) Inventor; and DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, (71) Applicant : SYAL, Rakesh, Richard, K. [CA/CA]; 2014- LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, 100 Wellesley Street East, Toronto, Ontario Canada M4Y SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, 1 H5 (CA). GW, KM, ML, MR, NE, SN, TD, TG). (74) Agent: SYAL, Rakesh, Richard, K.; 2014-100 Wellesley Published: Street East, Toronto, Ontario Canada M4Y 1 H5 (CA). — with international search report (Art. 21(3)) (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM,
0 0 (54) Title: EDIBLE OIL BLEND COMPOSITION FOR USE BY HUMANS WITH FOOD ALLERGIES & INTOLERANCES © (57) Abstract: A certified organic and certified non-GMO edible oil blend composition for humans with food allergies and intoler ances referred to as AF8-oil. AF8-oil comprises Avocado oil, Camelina oil, Chia Seed oil, Flaxseed oil, Hemp oil, Olive-Extra Vir gin oil, Rice Bran oil and Sacha Inchi oil. AF8-oil comprises 2.1 g/ml to 14.26 g/ml of protein. AF8-oil tests negative to allergens ¾ using Skin Prick Tests on humans. AF8-oil comprises 20% to 35% MUFAs, 8% to 15% SFAs, and 50% to 70% PUFAs wherein PUFAs is comprised of Omega 3 to Omega 6 ratio from 1 :1 to 1 :4. AF8-oil comprises vitamins E, K and beta-carotene; and com- prises phystosterols. AF8-oil which comprises enhanced antioxidant activity with an Oxygen radical absorbance capacity of 414.0 to 426.0 µιηοΐ Trolox equivalent/1 OOg and Total Phenol Content of 35.0 to 47.2 mg GAE/100 g (Gallic Acid Equivalent). AF8-oil S exhibits superior flavour and taste. To be used for preparing foods. EDIBLE OIL BLEND COMPOSITION FOR USE BY HUMANS WITH FOOD ALLERGIES & INTOLERANCES
Technical Field
[0001] The present invention relates to the field of an edible oil blend composition for preparing of food by humans with food allergies and intolerances while maintaining a healthy diet and good health.
Background of the Invention
[0002] An allergy is a hypersensitive disorder of the immune system. An allergy is one of four forms of hypersensitivity reactions and is called type I (or immediate) hypersensitivity. An allergy is caused by an adverse immune reaction to specific substances known as allergens.
[0003] Food allergy is an allergy caused by an allergic reaction to a substance ingested from food. In other words, a food allergy is an allergy wherein the allergen is a food component. In particular, a food protein, a food protein fragment or a food protein fraction may act as an allergen in food allergies. A food allergy should be distinguished from other adverse responses to food, such as food intolerance, pharmacological reactions and toxin-mediated reactions.
[0004] Food allergies are a growing health concern with dramatic increase in the last two decades. More than 1 million US Americans suffer from food allergies. That's 4% of the population. Between 6 and 8 per cent of Canadians have food allergies. In human terms, that means up to one in
13 Canadians is food allergic. The European Academy of Allergy and Clinical Immunology (EAACI) estimates more than 17 million people in Europe suffer from food allergies of which 3.5 million European sufferers are younger than 25 years old and the sharpest rise in food allergies is amongst children and young people.
[0005] Allergens are identified by the organism and tagged with Immunoglobulin E (IgE). The immune system subsequently recognizes the tagged allergens as harmful and consequently triggers an allergic reaction. In an individual suffering from a food allergy, a generally harmless food protein is mistakenly identified by the body's immune system as harmful. The food protein thus acts as an allergen and may therefore also be referred to as an allergenic protein. Although strictly speaking, the allergy is caused by the allergenic protein, food allergies are generally named after the food which comprises the allergenic protein. For example, peanut allergy is an allergy caused by peanut protein. [0006] Allergic responses caused by food allergy can range from mild responses, such as dermatitis, gastrointestinal and respiratory distress, to severe reactions, such as anaphylaxis, including biphasic anaphylaxis and vasodilation.
[0007] Anaphylaxis is a life threatening systemic condition, causing a constriction of the trachea, preventing breathing, and anaphylactic shock that could result in death. The number of severe and potentially life-threatening allergic reactions (anaphylaxis), due to food allergies, occurring in children is also increasing. The reasons for increased incidence of food allergies and associated anaphylaxis are poorly understood. Research suggests that food-related anaphylaxis might be under¬ diagnosed.
[0008] Delayed food allergy or food sensitivity also involves the immune system. Delayed food reactions may be associated with over 100 allergic symptoms and well over 150 different medical diseases. Symptoms of this type of food allergy may include: abdominal pains; aches and pains; acne; attention deficit hyperactivity disorder (ADHD); anxiety; arthritis; asthma; autism; bloating; celiac disease; chronic fatigue; chronic infections; constipation; depression; dizziness; eczema; edema; enuresis (i.e., bed wetting); fibromyalgia; fluid retention; gastrointestinal (GI) problems; hyperactivity; irritable bowel syndrome; itching; lethargy; loss of appetite; migraine; nausea; abnormal postmenstrual syndrome (P S); psoriasis; recurrent ear infections; recurrent sinus infections; rhinitis; sinusitis; skin rashes; stomach cramps; tension; urticaria; weight grain; weight loss; and wheezing.
[0009] The most common allergenic foods associated with IgE-mediated reactions on a worldwide basis are fish, peanuts, soybeans, milk, eggs, Crustacea, wheat, and tree nuts. These commonly allergenic foods account for >90% of food allergies. Peanut, tree nut, legumes and seed allergies, which tend to develop in childhood, usually are lifelong.
[00 10] Peanuts and tree nuts are some of the most common food allergies. Peanuts belong to the family of legumes (Fabaceae). Peanut allergies may be extremely severe. Proteins in tree nuts, including pecans, almonds, cashews, pistachios, pine nuts, and walnuts, are another widespread allergen that may also be extremely severe. Subjects suffering from tree nut allergy may be sensitive to one, or many, tree nuts. Furthermore, seeds, including sesame seeds and poppy seeds, may contain oils comprising a protein that can act as an allergen. [001 ] The prevalence of allergies to tree nuts, peanuts and seeds are considered significant health risks to the population and are classified as major or priority allergens by the European Union (EU), Canada and the United States. Food labeling regulations require that major or priority allergens be clearly identified on food labels. Nut allergy is the most common food to cause severe anaphylactic reactions.
[0012] TABLE 1 below shows the list of plant based sources classified as major or priority allergens by the USA, Canada and the EU
TABLE 1
Plant based Major or Priority Allergens
* Coconut is not a botanical nut; it is classified as a fruit, even though the Food and Drug Administration in the USA recognizes and classifies coconut as a tree nut.
[0013] TABLE 2 below Summarizes research on allergens of legumes, nuts and seeds TABLE 2: Summary of research on allergens of legumes, nuts and seeds
PLANT SOURCE SUMMARY OF RESEARCH FINDINGS REFERENCES
Tree Nuts (including but Tree nut and peanut oils may pose a threat J Allergy Clin Immunol not limited to almonds, to patients with allergy, depending on the 1997;99:502-6; Teuber SS et al., Arshad SH et al., Asero R et Brazil nuts, cashews, method of manufacture and processing. al., Beyer K et al., Birosel DM., hazelnuts, macadamia nuts, Even traces of residual Almond protein in Borja JM., Couturier P., pecans, pine nuts, Almond nut oil may pose a threat to Fleischer DM., Enrique E et pistachios or walnuts & patients with allergy. Cross-reactivity has al.,Goetz DW et al., Hansen KS Coconut ) been shown between Walnut, Sunflower et al., Maloney JM et al., seed and Peanut; between Peanut, Brazil Masthoff LJ et al., Moreno FJ et nut and Almond nuts and extracts; and al., Noorbakhsh R et al., Vocks E et al., Hasegawa M et al.. between pistachio and other tree nuts. Hide DW et al., Pinola A et al. Severe gastro-intestinal symptoms due to Robotham JM et al. Coconut oil and life-threatening systemic reactions, after consumption of Coconut has been demonstrated .
Peanuts Peanut oil, refined or unrefined ( cold Ben-Shoshan M et al., pressed) may contain peanut allergens. Flinterman AE et al., Garcia BE Some Peanut-allergic patients react to et al., Goetz DW et al., refined Peanut oil but not to unrefined Hoffman DR et al., Hourihane Peanut oil. Peanut contains up to 32 JO et al., Jacquenet S et al., different proteins, of which at least 18 have Maloney JM et al., Mondoulet L been identified as allergens, being capable et al., Moneret-Vautrin DA et of binding allergen specific IgE. Cross- al., Olszewski A et al., allergenicity has been demonstrated Saavedra-Delgado AM. between extracts of Peanut, Garden pea, Chick pea, and Soybean.
Seeds, Soybean & Other Anaphylaxis, Symptoms of skin and Asero R et al., Chiu JT et al., mucous membranes, Gastrointestinal Kanny G et al., Blamoutier J et symptoms and other allergic symptoms al., James C et al., Gangur V et have been reported in both Sesame seed and al., Vocks E et al., Ranee F et Sesame oil to as little as 2 to 3 ml, al. Codina R et al., Dominguez J Sunflower oil to as little as 5ml, and et al., Errahali Y et al., Figueroa mustard seed , rapeseed, grape seed and J et al., Foucard T et al., Fritsch soybeans, soybean oil & cottonseed. Cross- R et al., Gogolewski M et al., reactivity between allergens in Sesame and Hiemori M et al., Inomata N et allergens in other foods, including Hazel al., Jorro G et al., Malet A et al., nut, Rye, Kiwi, Poppy seed, Black walnut, Kalogeromitros DC et al., Cashew, Macadamia, Pistachio, and Maloney JM et al., Neering H et Peanuts, has been reported. Gluten sensitive al., Noyes JH et al., Paschke A patents have shown allergies to wheat germ et al., Porras 0 et al., Romano C oil. et al., Zitouni N et al. [0014] There is no cure for food allergies. Strict avoidance of food allergens and early recognition and management of allergic reactions to food are important measures to prevent serious health consequences.
[0015] Most plant based edible oils used for human consumption may be derived from a number of major or priority food allergens, including soybean, peanuts, tree nuts (i.e., almonds, Brazil nuts, cashews, hazelnuts, macadamia nuts, pecans, pine nuts, pistachios or walnuts & Coconut ), sesame and mustard seeds. Edible oils can also be derived from foods not classified as priority allergens such as sunflower, grape seed, cottonseed, wheat germ, pumpkin, palm, which still may present an allergen health risk.
[0016] Oils can be either Refined or Unrefined. Government health regulatory bodies in the USA, Canada, and EU have determined that ingredients derived from a food allergen but that has been processed in a way that removes the protein are not themselves considered to be food allergens. Highly refined oils may be derived from a priority food allergen source. However, because of the degree to which they have been refined, contain no protein or an amount of protein that is too small to pose an allergen risk. However, there is still no guarantee that an allergic reaction will not occur to refined oils. Olszewski et al., demonstrated the presence of allergenic proteins in unrefined and refined peanut oil. Ramazzotti M et al., in a study on protein content in refined seed oils concluded " it cannot be excluded that allergic patients may present specific IgE for number of proteins larger than that known, some of them still present in refined oils". Zitouni N et al., found that because of the presence of allergenic proteins, refined sunflower oil may pose a threat to people highly sensitized to sunflower seeds. The Joint Food Safety & Standards Group, Department of Health, London, United Kingdom recognized that the allergenicity of vegetable oils is highly dependent on the production processes used to extract and then refine the oils.
[0017] Refined oils may be produced using a number of different steps. The crude oil is generally produced by mechanically pressing the source seed or bean in expellers after a preheating step in indirectly heated conditioners. The crude oil can then be treated using a number of different refining steps. Degumming, neutralizing, bleaching and deodorizing are all steps that are used to treat the crude oil and produce what is then considered to be a highly refined oil. Solvent extraction is used to separate oil from seeds/beans where the principle is to employ a volatile liquid in which the oil is freely soluble. For example, it is common in the food industry to use the solvent hexane in crushers, and treat seed oils with sodium hydroxide and hydrochloric acid, and bleach the oils using clays to remove color pigments. The pre-processed seeds/beans are treated in a multistage counter-current process with solvent until the remaining oil content is reduced to the lowest possible level. Crushed seeds are subjected to temperatures of 230 °F to 356 °F ( 110 to 80 °C ) in a steam bath to start the extraction process.
[0018] Refining removes or damages many beneficial substances found in natural edible oils. High temperature processing causes the weak carbon bonds of unsaturated fatty acids, especially triple unsaturated linolenic acid, to break apart, thereby creating dangerous free radicals and damaging alpha linolenic acid (ALA). ALA is particularly susceptible to being damaged by excessive heat. For example, antioxidants, such as tocopherols, sterols, and polyphones which protect the body from the ravages of free radicals, are neutralized or destroyed by high temperatures and pressures (Prior et al., Gogolewski et al., Koski et al., Garcia A et al.). Wroniak et al., found that deodorised oils contained 1.1% trans isomers of fatty acids that been found to be harmful to human health, and contained 25% to 45% less total tocopherols than cold pressed oils. Likewise in other investigations (Kristott., Gogolewski et al.), Rafalowski, R et al., found that β - carotene, a precursor to Vitamin A and present in most cold pressed oils, was absent in refined oils. Heavy use of refined cooking oils in human diets is attributed to increased risk of cancer, inflammation and cardiovascular disease, and other negative health consequences.
[0019] Unrefined oils (also commonly referred to as "crude" oil) are most commonly produced using one of two methods referred to as "Expeller" or "Cold Pressed" . An expeller pressed oil is an oil that has come about by means of chemical free, mechanical or physical methods. The raw material (usually nuts, seeds and occasionally algae) is pressed, squeezed, and or crushed under high pressure until oil begins to seep out from the "parent material". This is the method that oil has been extracted for centuries worldwide. The temperature reached during this crushing and pressing process often depends on the hardness of the object being crushed. The harder the seed/nut the more friction, thus heat, is created. There is no external heat added in expeller pressed oils, only what is generated by the friction of material being pressed. Cold pressed oils are a form of expeller pressed oils. These oils follow the same guidelines of no chemicals used in the process and extracted only by mechanical or physical means. The difference being, that Cold Pressed oils cannot be brought to a temperature over 120 °F (approx. 49 °C). This method preserves all of the delicate phytochemicals that make the nut or seed oil truly beneficial. All of the nuances of flavor (if using for cooking) are preserved, and nutritional values, color, natural odor are all held intact. Europe and the State of California, USA, have rigorous standards in place when it comes to using the words "cold pressed" with oils. [0020] Cold-pressing is believed to be a better technique for retaining beneficial macronutrients such as antioxidants, vitamins E and K, phenolic compounds, phytosterols and avoiding damage to the essential fatty acids such as ALA in seed oils that might be lost by evaporation or chemically modified using conventional solvent extracting methods (Parker TD et al.).
[002 1] Allergy sufferers, particularly to tree nuts, and variety of seeds such as sesame, mustard, sunflower, soybeans and peanuts are left with very limited selection of oils to use that have not been demonstrated to pose allergen health risks.
[0022] TABLE 3 below lists the most oils generally available for human consumption. There are a total of 29 oils, of which 20 or 68.96% of the oils where allergen health risks have been reported indicated by and 9 oils or 31.03% have not been reported to pose an allergen health risk indicated by X . These 9 oils include Avocado, Camelina, Extra Virgin Olive, Chia Seed, Flax Seed, Hemp, Sacha Inchi, Safflower, and Rice Bran. Of these 9 oils 4 oils are relatively new to the consumer, and include Camelina, Chia Seed, Sacha Inchi, and Hemp. TABLE3
List of Common Available Oils for Human Consumption
Total = 29
* New oils not commonly known by consumers
Sources: United States Department of Agriculture Nutrient Database Release 27 & Other Sources
PUFA= Polyunsatured fatty acids; MUFA=Monounsaturated fatty acids; SFA=Saturated fatty acids GAE= Gallic Acid Equivalent / Vit E includes Tocopherols & Tocotrienols Phytosterols include Cholesterol, Stigmasterol, Campesterol, Beta-sitosterol and ∆5 - avenasterol X = Allergen risk to human health has not been reported. Allergen risk has been reported and poses allergen risk to human health / NA - Not applicable [0023] The health benefits of edible oils for human consumption is well understood and documented. Amongst many health benefits provided by these oils, some major benefits include providing PUFAs that are essential to human health. Two fatty acids include Omega- 6 in the form of linoleic acid (18:2n-6) and Omega -3 in the form of alpha-linolenic acid (18:3n-3). These two fatty acids are essential to human health because they cannot be synthesized de novo, therefore, they must be obtained from exogenous sources. These essential fatty acids are required by the brain and nervous system and for normal growth and vision development in infants . Deficiencies of essential fatty acids are usually characterized by dry flaky skin, decreased growth and wound healing, diarrhea, increased anemia, and an increased rate of infections . Aside from being nutritionally essential, linoleic acid has been considered a cholesterol lowering fatty acid for a number of years, and LDL (Low-density lipoprotein) is the primary cholesterol that it decreases. Using oleic acid as a baseline, it has been estimated that increasing linoleic acid intake lowers cholesterol about half as much as saturated fatty acids increase it. Both Alpha-linolenic acid and linoleic acid have also been shown to decrease LDL (Grundy S et al., Wijendran V et al.).
[0024] It is not only important to incorporate Omega-3 and Omega-6 in a healthy diet, but also consume these essential fatty acids in the correct ratio. Omega-6 fatty acids may compete with Omega-3 fatty acids for use in the body (Lands WE et al.) and therefore excessive intake of Omega- 6 can inhibit Omega-3. The ratio of Omega-3 to Omega-6 fatty acids preferably should be between 1:1 and 1:4 (Simopoulos AP). Instead, it is estimated that most people in the western world consume these fatty acids at a Ratio of Omega-3 to Omega-6 between 1:10 and 1:25, and are consequently unable to reap the benefits of Omega-3s (Simopoulos AP., Candela CG et al).
[0025] Antioxidants found in oils are very important dietary components. Nutrient antioxidants that are essential to the body include vitamin C and vitamin E. Cells contain many potentially oxidizable substrates such as polyunsaturated fatty acids (PUFAs), proteins, and DNA. Therefore, a complex antioxidant defence system normally protects cells from the injurious effects of endogenously produced free radicals as well as from species of exogenous origin such as cigarette smoke and pollutants. Should our exposure to free radicals exceed the protective capacity of the antioxidant defence system, a phenomenon often referred to as oxidative stress, then damage to molecules may occur. There is considerable evidence that disease causes an increase in oxidative stress; therefore, consumption of foods rich in antioxidants, which are potentially able to quench or neutralise excess radicals, may play an important role in modifying the development of such diseases. These vitamins protect the body from free radical damage among other functions. Antioxidant compounds found in most edible plant based oils include vitamin E ,carotenoids, phenolic and polyphenolic acids. Phenolic compounds have demonstrated to be powerful antioxidants. Individual phenolic compounds and foods containing phenolic compounds have shown strong abilities to significantly extend the presence of vitamin C when exposed to ascorbic acid oxidase (Cossins E et al.) and individual phenolic compounds have shown to be very effective free radical scavengers (Rice- Evans CA et al.). Many epidemiological investigations indicate that dietary antioxidants are associated with reducing the risk of chronic diseases including cardiovascular disease, cancer, and diabetes (Temple NJ et al.). Natural antioxidants, including vitamin E and phenolic compounds, are prevalent at significant levels in some seed oils. Vitamin E has also been shown to play a significant role in the formation of red blood cells and helps the body utilize Vitamin .
[0026] The generic term "vitamin E" is used to designate eight different compounds, comprised of four tocopherols -, β -, γ - and δ -. (Alpha, beta, gamma and delta) and four tocotrienols -, β -, γ - and δ -. (Alpha, beta, gamma and delta). Because humans and animals do not synthesize their own vitamin E, they primarily acquire tocopherols from plants, which are the only species capable of making vitamin E. y-tocopherol is often the most prevalent form of vitamin E in plant seeds and in products derived from them (McLaughlin PJ et al.). Vegetable oils such as corn, soybean, and sesame, and nuts such as walnuts, pecans, and peanuts are rich sources of y-tocopherol (McLaughlin PJ et al.). Because of the widespread use of these plant products, y-tocopherol represents 70% of the vitamin E consumed in the typical US diet (McLaughlin PJ et al.). In contrast, —tocopherol is the predominant form of vitamin E in most human and animal tissues, including blood plasma. In rats, —tocopherol concentrations are generally much higher than those of y- tocopherol (Clement M et al.). In humans, plasma —tocopherol concentrations are generally 4-10 times higher than those of y-tocopherol (Behrens WA et al.). Studies that report y-tocopherol concentrations in human tissues other than plasma are rare and mostly limited to adipose tissue (Handelman GJ et al.). However, Burton et al., reported that y-tocopherol constitutes as much as 30-50% of the total vitamin E in human skin, muscle, vein, and adipose tissue. Importantly, y- tocopherol concentrations in these tissues appear to be 20 0-fold greater than those in plasma (Burton et al) . Furthermore, y-tocopherol concentrations are substantially higher in human than in rodent tissues.
[0027] Beta-carotene is a carotenoid, which are substances that contain pigments responsible for color in foods, such as orange in carrots. Like Vitamin E, beta-carotene acts as an antioxidant, substances that deactivate free radicals. Among many other positive health benefits, carotenoids act as a precursor to vitamin A. Beta-carotene is thought to possess many positive health benefits and in particular helps prevent night blindness and other eye problems. It also effective in skin disorders, enhances immunity, protects against toxins and cancer formations, colds, flu, and infections. It is an antioxidant and protector of the cells and may slow the aging process. It is considered that natural Beta-Carotene aids in cancer prevention. It is important in the formation of bones and teeth. No vitamin overdose can occur with natural Beta-Carotene. It has been reported that beta carotene offers a notable measure of photoprotection to individuals with porphyria (Beta-Carotene helps to protect the eye and vision).
Vitamin A is necessary for a large number of metabolic functions similar to other vitamins. One of the major unique functions of vitamin A is its role in vision, especially dim-light vision. In third world countries where fruit and vegetables are not readily available, vitamin A deficiency is the leading cause of blindness in children. Vitamin A is also an important antioxidant that may play a role in the prevention of certain cancers.
[0028] Vitamin K helps with many important functions in your body. Vitamin helps the human body heal wounds, maintain blood vessels and keep bones healthy. Vitamin K may also help in preventing fractures (broken bones), especially in women after menopause. Observational studies have found a relationship between vitamin K and age-related bone loss (osteoporosis) (Feskanich D et al., Shearer MJ., Shearer MJ et al.). Even though major sources of vitamin are green, leafy- vegetables such as turnip greens, spinach, cauliflower, cabbage and broccoli, most people in Canada, USA and EU do not eat enough green leafy vegetables and about 76% tend to get their vitamin in the diet from vegetable oils. Vitamin K exists in two naturally occurring bioactive forms; phylloquinone (vitamin Kl) and menaquinone (vitamin K2). The phylloquinone is the most common form of vitamin K.
[0029] Phytosterols and phytostanols, also referred to as plant sterols and stanols, are common plant and vegetable constituents and are therefore normal constituents of the human diet. In human diets, vegetable oils are a major source of plant sterols but they can also be found in other plant- derived foods, such as nuts, cereals and legumes. Studies have shown that plant sterols mimic cholesterol in the small intestine and partly block cholesterol absorption. Therefore, regular consumption of a variety of foods containing plant sterols may result in the lowering of LDL- cholesterol levels, while having no detrimental effect on HDL-cholesterol levels (often referred to as good cholesterol). This would benefit those who want to lower their blood cholesterol levels, i.e., especially those who have moderate to high blood cholesterol. The importance of the role phytosterols and phytostanols play in lowering LDL- cholesterol levels has been approved by health regulatory agencies around the world and include, the European Union, Canada, USA, Australia, Switzerland, Norway, Iceland, Brazil, South Africa, Japan, Turkey and Israel. The most common phytosterols and phytostanols found in plant based edible oils are β —sitosterol, campesterol, stigmasterol, brassicasterol, and ∆5 —avenasterol.
[0030] Prior art includes numerous patents in the areas of but not limited to edible oil compositions and blends, on ratios of Omega 6 to Omega 3, uses of Omega 3, other oil compositions to treat particular health risks, Vitamin E and antioxidant activity. Examples of prior art include US Patent 2008/0206435 discloses oil composition for deep frying food; US 2005/03987(CA25586309 A1/EP1827123 A2) discloses oil composition having high concentration of poly-unsaturated fatty acids with better stability and minimum trans-fatty acids; WO2005020698 A l ( US 2007/0071872 / CN 1845678A /EP 1510133 Al) discloses oil composition having a balanced ratio of Omega 6 to 3 of 1:1; WO2012145339 A2 discloses oil composition having high efficacy ratio of omega 6 to omega 3; US 2010/0209564 A l discloses oil composition that reduces photoxidation and having a good flavor; US 201 1/0274790 A discloses nourishing oil composition for infants and young children; WO2004002234 Al discloses dietary extra-virgin olive oil with fortified Omega 3 increasing the amount of Omega 3 in olive oil; WO2004056348 discloses natural vitamin E composition with superior antioxidant potency; US 2014/0171674 A discloses functional food comprising high oleic canola oil for mitigating cardiovascular disease risk; and WO200191587 A2 discloses oil composition use thereof in reducing weight gain.
[0031] Many of the prior art blends compositions make little or no distinction between non-GMO (genetically modified organisms), organic and non-organic oils, the method of production used in making the oils, and oils that may be an allergen health risk. Where prior art has disclosed the method of production, in particular between refined, unrefined , GMO and non-GMO, organic and non organic such as US Patent 1992/5169669, US Patent 201 1/7902388 B2 and WO 2008058361, it is not clear why the oils or oil seeds are considered organic or non-GMO, and no distinction is made between oils or oil seeds that may present an allergen health risk. There is thus a need in the art to produce foods that are suitable for people with food allergies and intolerances. Accordingly, there is a need for an edible oil used in preparing food suitable for people with food allergies and intolerances. Further, this edible oil contains essential fatty acids omega 3 and omega 6 in the correct ratio of Omega 3 to Omega 6, vitamins E, K, beta-carotene, phytosterols and antioxidants so that health conscious consumers with food allergies may be able to obtain these dietary nutrients from one convenient source. Further that the edible oil be organic, non-GMO, and be produced in a manufacturing facility free of allergens. Further, there is a need for this oil to provide adequate amount per Tablespoon of each omega-3, vitamin E, Vitamin K, phytosterols. Further, this oil, that may be used every day, year after year, and tastes good with good flavor. Definitions
[0032] Hereinafter, the terms, "oil", "edible oil" and "vegetable oil" shall be used interchangeably and shall refer to "plant based oils" used for human consumption. Where "plant based oils" shall mean any of a group of liquid edible fats that are obtained from plant sources that include but not limited to nuts, seeds, fruits and flowers.
[0033] The term "edible" in the context of a fat-based composition means that said composition is suitable for use in mammalian, e.g., human, foods, dietary supplements and pharmaceutical preparations.
[0034] By "comprising" or "comprises" or "comprised" is meant including, but not limited to, whatever follows the word "comprising" or "comprises" or "comprised". Thus, use of the term "comprising" or " comprises" or "comprised" indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By "consisting of or "consists of or "consisted of is meant including and limited to, whatever follows the phrase "consisting of or "consists of or "consisted of. Thus, the phrase "consisting of or "consists of or "consisted of indicates that the listed elements are required or mandatory, and that no other element may be present. By "consisting essentially of or consists essentially of or "consisted essentially of is meant including any elements listed after the phrase , and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase "consisting essentially of or consists essentially of or "consisted essentially of indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending on whether or not they affect the activity or action of the listed elements.
[0035] Reference throughout this document to "embodiment", "another embodiment", "general embodiment', "one embodiment", "certain embodiments", and "an embodiment" or similar terms means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the appearances of such phrases or in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments without limitation. [0036] As used herein the phrase "adequate" amount refers to amount per Tablespoon or teaspoon from foods or per capsules, softgels, or millilitres or ounces per bottle, or any such unit that may comprise pharmaceuticals, food, beverages and nutraceuticals manufactured or supplemented with the oil blend composition described herein, for maintaining good health and diet in humans.
Brief Summary of the Invention
[0037] Many of the prior art blends compositions of edible oil for humans make little or no distinction between non-GMO (genetically modified organisms), organic and non-organic oils, the method of production used in making the oils, and oils that may be an allergen health risk. There is thus a need in the art to produce foods that are suitable for people with food allergies and intolerances. Accordingly, there is a need for an edible oil used in preparing food by people with food allergies and intolerances.
[0038] An edible oil blend composition for preparing food by humans with food allergies and intolerances, comprising of Avocado oil, Camelina oil, Chia Seed oil, Flaxseed oil, Hemp oil, Olive-Extra Virgin oil, Rice Bran oil and Sacha Inchi oil wherein each oil is unrefined, cold pressed, certified organic and certified non-GMO ( not genetically modified), manufactured in a facility free of allergens such as tree nuts, peanuts, sesame and mustard seeds, and soybean avoiding cross-contact with allergen contaminates. Hereinafter referred to as "AF8-Oil".
[0039] AF8-Oil comprises 1.0 to 2.5 wt.% of Avocado oil; 50 to 75% wt.% of Camelina oil; 3 to
15 to 20% wt.% ofchia seed oil; 5% wt.% of flaxseed oil; 2.5 wt.%; 1.0 to 2.5 wt.% of hemp oil; 1.0 to 2.5 wt.% of rice bran oil; 15.0 to 20.0 wt.% of sacha inchi oil; and 3.0 to 5.0 wt.% of extra virgin olive oil.
[0040] AF8-Oil contains 2.1 µg/ml to 14.26 ig/ of protein and tests negative to Skin Prick Tests for allergens on human subjects.
[0041] AF8-Oil comprises 20% to 35% of MUFAs, 8% to 15% of SFAs, and 50% to 70% of PUFAs, wherein 20% to 30% of total PUFAs is omega-6 and 30% to 45% is omega-3, ratio of omega-3 to omega-6 is the range of 1:1 to 1:4.
[0042] AF8-OU comprises 40.00mg/100g to 42.00 mg/lOOg of Vitamin E, 7.0 mg/lOOg to 9.0 mg/lOOg of Beta-Carotene, 22.0 ug/lOOg to 33.5 ug/lOOg of Vitamin K, and 533.0 mg/lOOg to 542.0 mg/lOOg of Phystosterols.
[0043] AF8-Oil exhibits enhanced antioxidant activity with ORAC values of 414.0 µιηοΐ Trolox equivalent/lOOg to 426.0 µηιοΐ Trolox equivalent/ lOOg and Total Phenol Content (TPC) of 35.0 mg GAE/100 g to 47.2 mg GAE/100 g ( GAE=Gallic Acid Equivalent). Detailed Description of the Invention
[0044] The present invention describes an edible oil blend composition for preparing of food by humans with food allergies and intolerances, particularly to , but not limited to, tree nuts, peanuts, sesame and mustard seeds, and soybean while maintaining a healthy diet and good health.
[0045] The present invention comprises a blend composition of eight edible oils, wherein each oil is certified to be organic, each oil is certified to be non-GMO, and each oil is manufactured and produced in an allergy free production facility. The present invention further tests negative for allergens on humans; comprises monounsaturated fatty acids (MUFAs); polyunsaturated fatty acids (PUFAs), where the PUFAs contain omega-3 and omega-6 fatty acids in a ratio of :1 to 1:4; low saturated fatty acids (SFA); vitamins E, vitamin , and phytosterols. The present invention further enhances antioxidant activity. The present invention further provides adequate amounts, per Tablespoon, of each omega-3, vitamin E, Vitamin , and phytosterols to maintain good health and diet of humans who may have food allergies and intolerances.
Oils Used in Making Oil Blend Composition
[0046] TABLE 4, below lists oils that have not been reported to pose allergen risk to human health. Of the oils listed in Table 4, 4 oils (Flax, Olive oil, Rice Bran, and Safflower) contain Omega-3, Omega-6, vitamin E, Vitamin k, and phytosterols. However, olive oil, rice bran and safflower oils are significantly deficient in Omega 3, while Flax is high in Omega 3. The disadvantage of these oils if taken by themselves is that it would result in a deficiency in either Omega 3 or Omega 6. The same deficiency in either Omega-3 or Omega-6 as well as deficiency in vitamin K would exist with the other 5 oils (Avocado, Camelina, Chia Seed, Hemp, Sacha Inchi). Chia Seed oil would also be deficient in vitamin E. Further, none of the oils in Table 4 have actually been tested for allergen risk in humans. TABLE 4
Potential Allergen Risk Free Oils
[0047] The present invention is related to an edible oil blend composition suitable for those with food allergies and intolerances wherein the edible oil blend composition, hereinafter referred to as "AF8-Oil", consists of 8 oils selected from the list of oils in Table 4 where the selected oils comprising "AF8-Oil" are Avocado, Camelina, Chia Seed, Flax Seed, Hemp, Olive Oil-Extra Virgin, Rice Bran and Sacha Inchi.
[0048] In an embodiment of the invention, AF8-Oil blend composition comprises Avocado oil, Camelina oil, Chia Seed oil, Flax Seed oil, Hemp oil, Olive Oil, Rice Bran oil and Sacha Inchi oil where each oil is certified Organic, each oil is certified non-GMO (genetically modified organisms), each oil is cold pressed and each oil is purchased from commercial sources using allergen free production methods.
[0049] To ensure the compliance to government regulatory organic standards in the USA, Canada and EU, each oil was certified by one or more of the following organic certification bodies: USDA Organic certified under the National Organic Program (NOP); and QAI - Quality Assurance International; Ecocert Canada and Ecocert Europe. To ensure compliance to non-GMO regulatory standards, each oil purchased was certified by either NON GMO PROJECT or Cert ID Non-GMO under EU Non-GMO regulatory standards. Organic and Non-GMO certifications are not limited to the certification bodies listed above. Oils may be certified by other bodies accredited to certify organic and non-GMO food products by their respective governments. [0050] Further each supplier(s) of oil (s) selected to supply the oil(s) for the present invention AF8- Oil was vetted to ensure that the oils are produced in a tree nut, peanut, sesame and mustard seeds, and soybean free manufacturing facility so to avoid any cross-contact of any allergens in the oils and the manufacturing facility is certified by an allergen free manufacturing or production program such as, but not limited to, the CAC (Certified Allergen Control) Program certified to minimize cross-contact of any allergen contaminants in the produced oil. Allergens, or proteins derived from allergenic foods, may be present in foods as the result of cross-contact during processing and handling. The term "cross-contact" describes the inadvertent introduction of an allergen into a product that would not intentionally contain that allergen as an ingredient. Cross-contact may occur when a residue or other trace amount of a food allergen is present on food contact surfaces, production machinery, or is air-borne, and unintentionally becomes incorporated into a product not intended to contain, and not labeled as containing, the allergen. Cross-contact may also result when multiple foods are produced in the same factory or on the same processing line as the result of ineffective cleaning, or may result from customary methods of growing and harvesting crops, as well as from the use of shared storage, transportation, or product equipment. Cross-contact of foods with allergens has been shown to lead to allergic reactions in humans (Jackson LS et al.).
AF8-OH Blend Composition
[005 1] In an embodiment of the invention AF8-OU blend composition comprises 8 edible oils: 1 to 2.5 wt.% of certified organic and certified non-GMO cold pressed Avocado oil; 50 to 75% wt.% of certified organic and certified non-GMO cold pressed Camelina oil; 3 to 5% wt.% of certified organic and certified non-GMO cold pressed flaxseed oil; 15 to 20% wt.% of certified organic and certified non-GMO cold pressed chia seed oil; 1to 2.5 wt.% of certified organic and certified non- GMO cold pressed of hemp oil; 1 to 2.5 wt.% of certified organic and certified non-GMO cold pressed of rice bran oil; 1 to 20 wt.% of certified organic and certified non-GMO cold pressed sacha inchi oil; 3 to 5 wt.% of certified organic and certified non-GMO cold pressed extra virgin olive oil, .
[0052] Fatty acid composition of the oil blend composition AF8-OU was determined by AOCS Official Method Ce lh-05. This method provides a gas-liquid chromatography (GLC) procedure for the determination of the fatty acid composition, including the trans fatty acid isomers of vegetable or non-ruminant animal oils and fats. The fatty acid methyl esters (FAME) of the sample are separated on a capillary gas chromatography column having a highly polar stationary phase, according to their chain length (CL), degree of unsaturation, and geometry and position of the double bonds. This method is specially designed to determine, by a single capillary GLC procedure, the levels of trans isomers, saturated fatty acid (SFA), cis- and /ram-monounsaturated fatty acid (MUFA), and cis- and trans polyunsaturated fatty acid (PUFA) levels in the same sample and same analysis.
[0053] In an embodiment of the invention AF8-Oil blend composition which comprises as percentage of total fatty acids 20% to 35% of monounsaturated fatty acids (MUFAs); as percentage of totally fatty acids 50% to 70% of polyunsaturated fatty acids (PUFAs); and as percentage of totally fatty acids 8% to 15% of saturated fatty acids (SFAs).
[0054] In an embodiment of the invention polyunsaturated fatty acids (PUFAs) comprises 20.0% to 30.0% as percentage of total PUFAs of omega-6 (n-6), and 30.0% to 45.0% as percentage of total PUFAs of omega-3 (n-3) wherein the ratio of omega-3 to omega-6 is in the range of : to 1:4.
Allergen Testing of AF8-OH
[0055] There are indications that the proteins present in oils may elicit allergic reactions in sensitive individuals. It is clear that the detection of proteins in oils are necessary to allow for a proper safety assessment. In a previous work, several methods of extraction and quantification of proteins were compared (Martin-Hernandez C et al.) and illustrates that the protein content of edible oils, both refined and unrefined, will vary by the extraction method used. Three methods of extraction were employed to determine the content of protein in the present invention AF8-Oil: Extraction with Acetone-Hexane (H); extraction with Acetone-Methanol (M) and Acetone extraction followed by Filtration (F).
Extraction with Acetone-Hexane (H). The isolation of proteins was performed using the method described by Paschke et al. for lecithin with some modifications. To 100 g of oil, 250 mL of Acetone-Hexane (1:1) was added. The mixture was shaken vigorously, kept for 1 h at 4°C, and shaken every 10 min. The mixture was then centrifuged, and the supernatant was discarded. The precipitate was washed twice with lOmL of Acetone-Hexane (1:1). After each washing, the mixture was centrifuged, and the supernatant was discarded.
Extraction with Acetone Methanol (M). This extraction was similar to the extraction of H (Extraction with Acetone-Hexane ) described above, but the precipitate was washed twice with 5 mL of Acetone Methanol (1:1) and once with 5 mL of Acetone.
Extraction with Acetone Followed by Filtration (F). The isolation of proteins was performed basically using the method described by Hidalgo et al.. To 100 g of oil, 250 mL of acetone was added. The mixture was shaken vigorously, kept forlhat4°C, and shaken everylO min and then filtered through Whatmanno.l filter paper using a Buchner funnel. Proteins from the paper were extracted with 5 mL of tetrahydrofurane and 5 mL of dioxane as described by Hidalgo et al..
The Protein in the present invention AF8-Oil was determined by Amino Acid Analysis. Proteins were quantified by using amino acid analysis, with a Hitachi L-8500 system (Tokyo, Japan). The method corresponds to AACC method 07-01 . Dried precipitates obtained from 100 g of oil were dissolved in 10 mL of 6 M hydrochloric acid, and nitrogen was introduced for 2 min. The solution was hydrolyzed in an oven for 24h a tl 10°C. The hydrolyzed sample was filtered into a 50 mL volumetric flask and made up to the mark with deionized water; afterward, this solution was evaporated. The residue was then dissolved in 2 mL of 0.02 M hydrochloric acid and filtered through a membrane filter before injection (75 µ ) on the amino acid analyzer.
[0056] TABLE 5 below shows the results of Protein Content in AF8-OU after Extraction of Lipids with Methods H, M, and F and Measured by Amino Acid Analysis
TABLE 5
Protein Content in AF8-Oil
a Mean values and standard deviations of ten replicates.
[0057] In an embodiment of the invention AF8-OU the protein content measured in / ml) may be in the range of values in the amounts of 2.1, 2.3, 2.5, 6.34, 6.7, 7.06, 12.34, 13.3, 14.26.
Skin Prick Test
[0058] Skin prick testing (SPT) - the primary mode of skin testing for immediate IgE mediated allergy. It is widely practiced, carries very low (but not negligible) risk of serious side effects to human subjects and provides high quality information (also called prick skin testing or PST). The SPT was used for detecting IgE-mediated food allergy to the present invention AF8-OIL in human subjects. 45 human subjects with past histories of food allergies were selected for the study with 10 healthy, non-atopic individuals serving as controls. Of the 45 allergic histories, 26 or 58% were female, and 14 or 42% were male. The age of the female group was from 8 years old to 18 years old, and the age of the male group was from 7 years old to 16 years old. The human subjects had a history of allergic symptoms to tree nuts (n=10), peanuts (n=l 5), sesame seed (n=5), mustard seed (n=3), sunflower, grapeseed, rapeseed (n=3), soybean n(=6), and olives (n=3).
The recommended method of prick testing includes the appropriate use of specific allergen extracts, positive and negative controls, interpretation of the tests after 15- 20 minutes of application, with a positive result (positive to allergens) defined as a weal greater than or equal to 3 mm diameter. If there is a positive reaction to the allergen a small lump (called a weal) will come up and reach its maximum size within 15 minutes (this weal can be red and itchy) an example is illustrated in Figure
1.
An extract of AF8-OU was created for administrating the SPTs. A prick-to-prick technique was utilized, i.e., first pricking the fresh food, in this case the AF8-OU extract, with the lancet and then pricking the skin, to test for sensitization to allergens.
The results of the SPTs were negative to AF8-Oil. 30 of the 45 subjects or 66.7% showed zero weals or lumps, 1 out of the 45 or 33.3% showed weals or lumps less than 3 mm diameter to AF8- Oil, indicating that AF8-OU is suitable for use by humans who have food allergies, particularly allergies to tree nuts, variety of seeds such as sesame, mustard, and sunflower, soybeans and peanuts. As a result, the present invention is suitable for use by humans who have food allergies and intolerances.
[0059] FIGURE 1 below illustrates an example of a Skin Prick Test where reaction to an allergen shows up as a small lump (called a weal). FIGURE 1
An Example of a Skin Prick Test (SPT)
[0060] In an embodiment of the present invention AF8-Oil, AF8-Oil tests negative to Skin Prick Tests (SPTs) on humans who have a history of food allergies, particularly allergies to tree nuts, sesame, and mustard seeds, sunflower, soybeans and peanuts.
Vitamin £ & β-carotene
[0061] Analyses of Vitamin E in the present invention, AF8-Oil was conducted by HPLC (High- performance liquid chromatography) , using chromatographic conditions: Si60 LiChrosorb column, 5 um; 250 x 4 mm; fluorescence detection (290 nm excitation and 330 nm emission); mobile phase hexane / isopropanol / acetic acid (99.33: 0.6: 0.07) with a flow rate of 1 mL min
Carotenes content in the present invention, AF8-Oil, was analysed by Ultraviolet-Visible (UV-vis) spectrophotometer at 446 nm using MPOB test method. The sample was homogenized and weighed to the nearest ±0.0001 g into a 25 mL volumetric flask. The sample was dissolved with n-hexane and diluted to the mark. The solution was transferred into a 1 cm quartz cuvette and the absorbance was measured at 446 nm against n-hexane.
[0062] TABLE 6 below shows the amounts of Vitamin E and β-carotene observed in AF8-Oil TABLE 6
Vitamin E & β-carotene for AF8-CM1 ( Mean values and std deviations with 5 replicates)
[0063] In an embodiment of the present invention AF8-Oii, comprises 46.00 mg/lOOg to 48.00 mg/lOOg of total vitamin E, wherein total vitamin E comprises 40.0 mg/lOOg to 42.0 mg/lOOg of total tocopherols and 5.0 mg/lOOg to 7.0 mg/lOOg of total tocotienols, wherein total tocopherols comprises 19.0 mg/lOOg to 21.0 mg/lOOg of a-tocopherols; 1 .0 mg/lOOg to 13.0 mg/lOOg of
β - tocopherol; 8.0 mg/100g to 10.0 mg/lOOg of γ - tocopherols; 1.0 mg/lOOg to 3.1 mg/lOOg of a - tocotienols and 2.0 mg/lOOg to 9.0 mg/lOOg of γ —tocotienols.
[0064] n an embodiment of the present invention AF8-OU, comprises 7.0 mg/lOOg to 9.0 mg/lOOg of total β —carotene.
Vitamin K
[0065] Extracted samples were quantified by HPLC (High-performance liquid chromatography) using C I 8 column (250 mm x 4.6 mm, 5.0 µ particle size). For the intention, 1 aliquot of sample was injected via auto sampler (WISP Model 710). The column temperature was maintained at 40°C. The mobile phase comprised of dichloromethane (100 mL), methanol (900 mL), zinc chloride (1.37 g), sodium acetate (0.41 g) and acetic acid (0.30 g). The flow rate was adjusted at 1 mL/min. Quantification of vitamin K was carried out using UV/vis detector (model 481) at 249 nm.
Concentration of Vitamin in the present invention AF8-Oil, was found to be a mean of 27.57 ± 5.5 (ug/lOOg) with five replicates.
[0066] In an embodiment of the present invention, AF8-Oil is comprised 22.0 ug/lOOg to 35.0 ug/lOOg of Vitamin K.
Phytosterols
[0067] Sterols were identified and quantified by liquid chromatography with mass spectrometry detection in positive APCI (atmospheric pressure chemical ionisation) mode. The samples were saponified by refluxing with 2 N ethanolic KOH, and the non-saponificable fraction was extracted with diethyl ether. This fraction was subjected to thin layer chromatography (TLC) on silica gel plates and then the band of sterols was isolated and extracted with methanol. Sterols were quantified by LC-MS, on a Waters Atlantis 5 µ η dC 2, l 150 mm column with a gradient of acetonitrile / water (0.01% acetic acid) at a flow of 0.5 mL min ; column temperature 30 °C. Concentration of the different sterols present in the oil was expressed as mg/100 g of oil. Cholesterol was the reference sterol used for determination of the response factor.
[0068] TABLE 7 below shows Phytosterols in AF8-OU
TABLE 7
Phytosterols in AF8-Oil (mg/lOOg) ( Mean values and std deviations with 5 replicates)
SITO = β —sitosterol, CAMP=campesterol, STIG= stigmasterol, BRASS= brassicasterol, and ∆5 - AVE = ∆ 5 - avenasterol.
[0069] In an embodiment of the present invention, AF8-Oil comprises 530.0 mg/lOOg to 542.0 mg/lOOg of Total Phytosterols, wherein total phytosterols comprises 281.0 mg/lOOg to 290.0 mg/lOOg of β -sitosterol; 186.0 mg/lOOg to 195.0 mg/lOOg of campesterol; 21.0 mg/lOOg to 30.0 mg/lOOg of stigmasterol; 23.0 mg/lOOg to 32.0 mg/lOOg of brassicasterol; and 3.0 mg/lOOg to 12.0 mg/lOOg of ∆5 —avenasterol.
Comparing AF8-OH to Potentially Allergen Risk Free Edible Oils
[0070] TABLE 8 below compares the present invention AF8-Oil to the Potentially Allergen Risk Free Oils TABLE 8 Comparison of AF8-OU to Other Potential Allergen Risk Free Oils
^Allergen risk proven not to pose health risk through Skin Prick Test on humans / X = Allergen risk to human health has not been reported
Antioxidant Activity
[0071] Total Phenols Content (TPC) was determined by a calibration curve of gallic acid in methanol was performed in the concentration range of 0.04-0.40 mg mL-1 . The solutions for the spectrophotometric analysis were performed as follows: in a 50 ml volume flask 1 mL of a standard solution of gallic acid, 6 mL of methanol, 2.5 mL of the Folin-Ciocalteau reagent, 5 mL of 7.5% Na2C03 were added, reaching the final volume with deionized water. The solutions were stored overnight and the spectrophotometric analysis was performed at λ= 765 nm. The TP determination was performed as follows: 2.5 g of the oil samples were diluted with 2.5 mL of nhexane and extracted three times by 5 minutes centrifugation (5000 rpm) with CH30H/H20 (80:20 v/v) extract. The extract was added to 2.5 mL Folin-Ciocalteau reagent, 5 mL of Na2C03 (7.5%), in a 50 mL volume flask reaching the final volume with deionized water. The samples were stored overnight and the spectrophotometric analysis was performed at λ= 765 nm. Gallic acid was used as the reference standard, and the results were expressed as mg of gallic acid equivalents and the total phenol content of each oil was subsequently calculated as gallic acid equivalent (mg GAE/lOOg).
[0072] OPvAC assay. Oxygen Radical Absorption Capacity (ORAC) assay measures the degree of inhibition of peroxy-radical-induced oxidation by the compounds of interest in a chemical milieu. It measures the value as Trolox equivalents and includes both inhibition time and the extent of inhibition of oxidation. The assay has been used to measure the antioxidant activity of foods. In addition to the ORAC assay, other common measures of antioxidant capacity (AC) include ferric ion reducing antioxidant power (FRAP) and trolox equivalence antioxidant capacity (TEAC) assays. These assays are based on discrete underlying mechanisms that use different radical or oxidant sources and therefore generate distinct values and cannot be compared directly. The ORAC assay is considered by some to be a preferable method because of its biological relevance to the in vivo antioxidant efficacy . Since antioxidant compounds with dissimilar chemical structures interact with different free radical sources, the relationship between any two AC methods will be quite low if considered across all foods. Thus, it is not possible to develop a mathematical relationship between two methods across a wide spectrum of foods. Like the content of any food component, AC values will vary due to a wide array of factors, such as cultivar, growing conditions, harvesting, food processing and preparation, sampling, and analytical procedures. For example, the ORAC assay was compared to other assays and the ORAC assay seems to provide a better correlation to the antioxidant capacity. Therefore, the ORAC assay method provides a valuable tool with which a researcher can quickly determine the value of a particular food, where increased potency and reduced cost are desired.
[0073] The method to establish an ORAC value of the present invention, AF8-Oil, method of Cao et al., was used with slight modifications as suggested by Ninfali, et al.. Peroxyl radicals were generated with AAPH, and B-PE was used as a detector of radical activity. When the sample had exhausted its capacity to trap peroxyl radicals, B-PE became the target of the radicals and rapidly lost its fluorescence. The area under the curve of fluorescence decay was proportional to the antioxidant capacity of the sample, and a comparative evaluation with Trolox was performed. The final reaction mixture for the assay was prepared in 10-mm-wide quartz cuvettes as follows: 1600
οί 0.04 µΜ B-PE in 0.075 M Na-K phosphate buffer pH 7.0, 200 of diluted phenolic extract, or 200 of 50 µΜ Trolox. A blank with 200 µ of methanol, diluted 1:20 (vol/vol) with 0.075 M Na-K phosphate buffer, pH 7.0, was also used; but, this blank did not differ from the one performed with the phosphate buffer alone. The mixture was kept at 37°C in the dark, and the fluorescence was read every 5 min at 565 nm emission and 540 nm excitation, using a PerkinElmer (Norwalk, CT) LS-5 spectrophotometer. When stability was obtained, the reaction was started with the addition of 200 µ of 40 mM AAPH and the fluorescence was measured at 37°C every 5 min until zero fluorescence was detected and calculated as µιηο Trolox equivalent/ 100 g. One of the benefits of using the ORAC method to evaluate substances' antioxidant capacity is that it takes into account samples with and without lag phases of their antioxidant capacities. [0074] TABLE 9 below shows the results of Antioxidant activity of the present invention, AF8-Oil.
TABLE 9 Antioxidant Activity of AF8-Oil
[0075] In an embodiment of the present invention, AF8-Oil shows enhanced antioxidant activity wherein the total ORAC values are 414.4 to 425.6 (µ οΐ Trolox equivalent/ 100 g) and the Total Phenol Content (TPC) is 35 to 47.1 (mg GAE/100 g ).
Adequate Amount
[0076] Most people tend to prepare food using a Tablespoon of oil. Consequently, food and nutritional labels on bottles of cooking oil sold in most retail outlets are labeled per Tablespoon of serving of the oil. Where tablespoon is generally used as a measure of volume in cooking or in consuming pharmaceuticals, food, beverages and nutraceuticals.
In most regions (including the United States, Canada, Japan, New Zealand, South Africa, United Kingdom and the Republic of Korea), one Tablespoon equals three teaspoons. In these regions, one
Tablespoon is approximately 15 mL, which is about V2 US fl oz. In Australia, a Tablespoon equals approximately 20 ml which is about ¾ fl. oz or approximately 4 teaspoons.
In some countries, a teaspoon (occasionally "teaspoonful") is a unit of volume, especially widely used in cooking recipes, generally preparing food, nutraceuticals and pharmaceutical prescriptions . In English it is abbreviated as tsp. or, less often, as t., ts., or tspn. The abbreviation is never capitalized because a capital letter is customarily reserved for the larger tablespoon ("Tbsp.", "T.", "Tbls.", or "Tb."). In German and Dutch teaspoon is abbreviated TL, for Teeloffel and Theelepel respectively.
[0077] In an embodiment of the present invention AF8-Oil, based on a 250 ml of AF8-OH, one Tablespoon, where one Tablespoon measures 14 ml to 20 ml, comprises, but not limited to, each of the following: Omega 3 in the amount of 3.0 grams to 8.0 grams; Omega 6 in the amount of 2.0 grams to 4.0 grams; Saturated Fatty Acids (SFAs) in the amount of 0.5 grams to 2.0 grams; Monounsaturated Fatty Acids (MUFAs) in the amount of 2.0 grams to 5.0 grams; Vitamin E (Alpha-Tocopherol) 4 milligrams to 8 milligrams; Vitamin in the amount of 2.0 micrograms to
6.0 micrograms ( ) ; and Phytosterols in the amount of 10.0 milligrams to 30.0 milligrams.
Taste and Flavour
[0078] Evaluation was conducted by 40 consumers. Using the 9-point Hedonic Scale ( Like Extremely being 1 and Dislike Extremely being 9) the consumers were asked to rank each oil according to flavor (Like the taste extremely being 1 to dislike the taste extremely being 9); Texture ( Extremely light being 1 to Extremely Heavy being 9); Oily or Greasy Aftertaste (Extremely Least
Greasy Aftertaste being 1 to Extremely Most Greasy Aftertaste being 9 ). Finally, the 40 consumers were asked to rank their preference for each of the oils. Following the 9-point Hedonic Scale the consumers ranked their preference for each oil (Extremely Preferred being 1 and Extremely Not Preferred being 9).
[0079] TABLE 10 below show the results of the evaluation by consumers of the taste and flavor of the present invention, AF8-OU. TABLE 10
Evaluation of Consumer Taste and Flavor Preference
[0080] In an embodiment of the invention, AF8-OU has excellent flavor and texture than any one single oil comprising the AF8-Oil blend composition, wherein the edible oil blend composition of the present invention, AF8-Oil, is strongly preferred by consumers over any one single oil comprising the AF8-Oil blend composition.
[0081] Any reference to claim elements in the singular, for example, using the articles "a" "an" or "the," is not to be construed as limiting the element to the singular. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the following claims and the principles and novel features disclosed herein. Patent Literature
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What is claimed is:
1. An edible oil blend composition for preparing food by humans with food allergies and intolerances, comprising of Avocado oil, Camelina oil, Chia Seed oil, Flaxseed oil, Hemp oil, Olive-Extra Virgin oil, Rice Bran oil and Sacha Inchi oil wherein each oil is unrefined, cold pressed, certified organic and certified non-GMO ( not genetically modified), manufactured in a facility free of allergens such as tree nuts, peanuts, sesame and mustard seeds, and soybean to avoid cross-contact with allergen contaminates.
2. An oil blend composition of claim 1 which comprises 1.0 to 2.5 wt. of Avocado oil; 50 to 75% wt.% of Camelina oil; 3 to 5% wt.% of flaxseed oil; 2.5 wt.%; 15 to 20% wt.% of chia seed oil; 1.0 to 2.5 wt.% of hemp oil; 1.0 to 2.5 wt.% of rice bran oil; 15.0 to 20.0 wt.% of sacha inchi oil; and 3.0 to 5.0 wt.% of extra virgin olive oil.
3. An oil blend composition of claim 1 which comprises 2.1 g/ml to 14.26 g/ml of protein.
4. An oil blend composition of claim 1 which tests negative to Skin Prick Tests (SPTs) on humans with a history of food allergies, such as allergies to tree nuts, variety of seeds such as sesame, mustard, and sunflower seeds, soybeans and peanuts, wherein the SPT results show zero weals or weals less than 3 mm.
5. An oil blend composition of claim 1, which comprises as percentage of total fatty acids 20% to 35% of monounsaturated fatty acids (MUFAs).
6. An oil blend composition of claim 1, which comprises as percentage of total fatty acids 8% to 5% of saturated fatty acids (SFAs).
7. An oil blend composition of claim 1., which comprises as percentage of total fatty acids 50% to 70% of polyunsaturated fatty acids (PUFAs).
8. An oil blend composition of Claim 7, wherein polyunsaturated fatty acids (PUFAs) comprises 20.0% to 30.0% as percentage of total PUFAs of omega-6 (n-6), and 30.0% to 45.0% as percentage of total PUFAs of omega-3 (n-3) wherein the ratio of omega-3 to omega-6 is in the range of :1 to 1:2. 9. An oil blend composition of Claim 1 which comprises 46.0 mg/lOOg to 48.0 mg/lOOg of vitamin E.
10. An oil blend composition of Claim 9 wherein total vitamin E comprises 40.00mg/100g to 42.0 mg/lOOg of total tocopherols, and 5.0 mg/lOOg to 7.0 mg/lOOg of total tocotienols.
11. An oil blend composition of claim 10 wherein total tocopherols comprises 19.0 mg/lOOg to 21.0 mg/lOOg of a-tocopherols;1 1.0 mg/lOOg to 13.0 mg/lOOg of β - tocopherols, ; and 8.0 mg/ lOOg to 10.0 mg/100 of - tocopherols.
12. An oil blend composition of claim 0 where total tocotienols comprises 1.0 mg/lOOg to 3.1 mg/lOOg of a- tocotienols, and 2.0 mg/lOOgto 4.1mg/100g of γ - tocotienols.
13. An oil blend composition of claim 1 which comprises 7.0 mg/lOOg to 9.0 mg/lOOg of β - carotene.
14. An oil blend composition of claim 1 which comprises 22.0 g/100g to 33.5 g/100g of Vitamin K.
15. An oil blend composition of claim 1 which comprises 533.0 mg/lOOg to 542.0 mg/lOOg of total phystosterols.
16. An oil blend composition of claim 15 wherein total phystosterols comprises 281.0 mg/lOOg to 290.0 mg/lOOg of/? -sitosterol; 186.0 mg/lOOg to 195.0 mg/lOOg of campesterol; 21.0 mg/lOOg to 30.0 mg/lOOg of stigmasterol; 23.0 mg/lOOg to 32.0 mg/lOOg of brassicasterol; and 3.0 mg/lOOg to 12.0 mg/lOOg of ∆ 5 - avenasterol.
17. The oil blend composition of claim 1 which comprises enhanced antioxidant activity with ORAC (Oxygen radical absorbance capacity) of 414.0 µ ο Trolox equivalent/ lOOg to 426.0 µ ο
Trolox equivalent/1 OOg and Total Phenol Content (TPC) of 35.0 mg GAE/100 g to 47.2 mg GAE/100 g (Gallic Acid Equivalent).
18. The oil blend composition of claims 1 based on a 250 ml of said oil blend composition wherein one Tablespoon measures 14 ml to 20 ml, may contain but not limited to each of the following per Tablespoon: 3.0 grams to 8.0 grams of Omega 3; 2.0 grams to 4.0 grams of Omega 6; 0.5 grams to 2.0 grams of Saturated Fatty Acids (SFAs); 2.0 grams to 5.0 grams of Monounsaturated Fatty Acids (MUFAs) ; 4 milligrams to 7 milligrams of Vitamin E (Alpha-Tocopherol); 2.0 micrograms ^g) to 6.0 micrograms ^g) of Vitamin K; and 10.0 milligrams to 30.0 milligrams of Phytosterols. International application No. INTERNATIONAL SEARCH REPORT PCT/CA2015/000441
A . CLASSIFICATION OF SUBJECT MATTER IPC: A23D 9/00 (2006.01) , 425 ) 9/007 (2006.01) , 4 77 57/722 (2006.01) , 4 7 57/202 (2006.01) , A61K 31/355 (2006.01) , A61K 31/56 (2006.01) (more IPCs on the last page)
According to International Patent Classification (IPC) or to both national classification and IPC
B . FIELDS SEARCHED
Minimum documentation searched (classification system followed by classification symbols) IPC (2006.01): A23D 9/00, A23D 9/007, A61K 31/122, A61K 31/202, A61K 31/355, A61K 31/56, A61K 31/575, A61K 36/00, A61P 3/02, CUB 1/00
Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched
Electronic database(s) consulted during the international search (name of database(s) and, where practicable, search terms used) Questel-Orbit (FamPat), CIPO Library Discovery Tool Keywords: oil*, blend*, mix*, combin*, antioxid*, orac*, synerg*, antagon*, avocado*, Persea*, camelina*, false_flax*, chia*, Salvia*, flax linseed*, linum*, hemp*, cannabis*, olive*, olea*, rice*, oryza*, sacha*, inchi*, Plukenetia*, volubilis*, mountain_peanut*, inca_peanut*
C . DOCUMENTS CONSIDERED TO BE RELEVANT Category' Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.
Y WO 2015/087277 A l (PICCARDI) 18 June 2015 (18-06-2015) 1, 3-16, 18 * page 8, lines 9-17 *
Y US 2013/026 1183 A l (BHAGAT) 3 October 2013 (03-10-2013) 1, 3-16, 18 * paragraphs 0066, 0123, 0143 *
Y PARKER et al., "Fatty Acid Composition and Oxidative Stability of Cold-pressed Edible Seed 1, 3-16, 18 Oils". Journal of Food Science, 2003, Vol. 68(4), pp. 1240-1243. * Introduction *
A WANG et al., "Synergistic, Additive, and Antagonistic Effects of Food Mixtures on Total Antioxidant Capacities". Journal of Agriculture and Food Chemistry, 201 1, Vol. 59, pp. 960-968.
Further documents are listed in the continuation of Box C. (" See patent family annex.
Special categories of cited documents: later document published after the international filing date or priority 'A' document defining the general state of the art which is not considered date and not in conflict with the application but cited to understand to be of particular relevance the principle or theory underlying the invention "E earlier application or patent but published on or after the international 'X' document of particular relevance; the claimed invention cannot be filing date considered novel or cannot be considered to involve an inventive 'V document which may throw doubts on priority claim(s) or which is step when the document is taken alone cited to establish the publication date of another citation or other Ύ document of particular relevance; the claimed invention cannot be special reason (as specified) considered to involve an inventive step when the document is Ό document referring to an oral disclosure, use, exhibition or other means combined with one or more other such documents, such combination being obvious to a person skilled in the art document published prior to the international filing date but later than document member of the same patent family the priority date claimed
Date of the actual completion of the international search Date of mailing of the international search report 8 March 2016 (08-03-2016) 22 March 2016 (22-03-2016)
Name and mailing address of the ISA/CA Authorized officer Canadian Intellectual Property Office Place du Portage I, CI 14 - 1st Floor, Box PCT Cara-Lynn Baas (819) 994-7568 50 Victoria Street Gatineau, Quebec K1A 0C9 Facsimile No.: 819-953-2476 Form PCT/ISA/210 (second sheet ) (January 2015) Page 2 of 5 International application No. INTERNATIONAL SEARCH REPORT PCT/CA2015/000441 C (Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT
Category' Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.
A WO 201 1/049969 A2 (PARKER) 28 April 201 1 (28-04-201 1) * entire document*
Form PCT/ISA/210 (continuation of second sheet) (January 2015) Page 3 of 5 INTERNATIONAL SEARCH REPORT International application No. Information on patent family members PCT/CA2015/000441
Patent Document Publication Patent Family Publication Cited in Search Report Date Member(s) Date
WO201 5087277A1 18 June 201 5 ( 18-06-201 5) WO201 5087277A1 18 June 201 5 ( 18-06-201 5) FR301 4688A1 19 June 201 5 ( 19-06-201 5)
US201 3261 183A1 03 October 201 3 (03-1 0-201 3) US201 3261 183A1 03 October 201 3 (03-1 0-201 3) AU201 13 15835A1 02 May 201 3 (02-05-201 3) AU201 13 15835B2 04 February 201 6 (04-02-201 6) CA281 4053A1 19 April 201 2 ( 1 9-04-201 2) CN1 03260437A 2 1 August 201 3 (21 -08-201 3) CO6700861A2 28 June 201 3 (28-06-201 3) EA201 390438A1 29 November 201 3 (29-1 1-201 3) EP26271 97A2 2 1 August 201 3 (21 -08-201 3) IL225574D0 27 June 201 3 (27-06-201 3) JP201 3541 108A 07 November 201 3 (07-1 1-201 3) KR201 301 4071 5A 24 December 201 3 (24-1 2-201 3) MX201 3003985A 0 1 May 201 4 (01 -05-201 4) SG1 89855A1 28 June 201 3 (28-06-201 3) SG1 0201 600295UA 26 February 201 6 (26-02-201 6) WO201 2051 591 A2 19 April 201 2 ( 1 9-04-201 2) WO201 2051 591A3 19 July 201 2 ( 19-07-201 2)
WO201 1049969A2 28 April 201 1 (28-04-201 1) WO201 1049969A2 28 April 201 1 (28-04-201 1) WO201 1049969A3 2 1 July 201 1 (21 -07-201 1) WO201 1049969A9 09 September 201 1 (09-09-201 1) CN1 02667469A 12 September 201 2 ( 12-09-201 2) JP201 3508724A 07 March 201 3 (07-03-201 3) KR201 20099688A 11 September 201 2 ( 1 1-09-201 2) US201 1136245A1 09 June 201 1 (09-06-201 1)
Form PCT/ISA/210 (patent family annex ) (January 2015) Page 4 of 5 International application No. INTERNATIONALSEARCH REPORT PCT/CA2015/000441
A61K 31/575 (2006.01) , A61K 36/00 (2006.01) , A61P 3/02 (2006.01) , CUB 1/00 (2006.01)
Form PCT/ISA/210 (extra sheet) (January 2015) Page 5 of 5