CLIAMS:
1. A synergistic premix composition for addition to an edible oil for enhanced stability during storage, in use in frying or other cooking methods, and in vivo after consumption of the food matrix, comprising:
a) ascorbyl palmitate/ stearate,
b) tertiary butyl hydroquinone (TBHQ),
c) tocopherol(s), and
d) dimethylpolysiloxane (DMPS).

2. The composition of claim 1, comprises natural and synthetic tocopherols.

3. The composition of claim 1, comprises 200 – 300 ppm of ascorbyl palmitate; 100 – 200 ppm of tertiary butyl hydroquinone; 300 – 1000 ppm of mixed tocopherols; and 1 – 10 ppm of DMPS.

4. An edible oil comprising a premix composition of claim 1, which may be selected from the group comprising of coconut oil, soybean oil, peanut oil, safflower oil, rapeseed oil, cottonseed oil, corn oil, rice bran oil, sunflower seed oil, sesame oil, palm oil, mustard oil, canola oil, high oleic oils, other vegetable oils and combinations thereof, either in their original state or hydrogenated and inter-esterified forms.

5. An edible oil comprising a premix composition of claim 1 may comprise one or more food grade additives such as vitamins, minerals or nutrients.
,TagSPECI:FIELD OF THE INVENTION
The present invention relates to the field of edible oils, especially edible oils for household use and for the food industry as ingredients in fried, baked or cooked food products and frying oils for room temperature stored fried food products. In particular, the present invention relates to a synergistic premix comprising an anti-oxidant combination for addition to an edible oil composition wherein the oil for enhanced stability during shelf life, frying and in vivo.

BACKGROUND OF THE INVENTION
Edible oils are an essential part of the human diet. They are recognized for their use in food preparation and as sources of essential fatty acids and micronutrients that are required by the human body. Edible oils derived from sources such as seeds and nuts, are fatty acid glyceryl esters. The major vegetable edible oils are rice bran oil, safflower oil, coconut oil, soybean oil, palm oil, rape seed oil, olive oil, sunflower oil, cottonseed oil, ground nut oil and high oleic oils like safflower, sunflower and canola oils. Edible oils are generally classified according to the relative proportions of the fatty acid constituents thereof, which may be saturated, monounsaturated or polyunsaturated. Saturated fatty acids, such as lauric, palmitic, stearic and arachidic acid, are free of doubly bonded carbon. A monounsaturated fatty acid, such as oleic acid, contains a single double bond, whereas polyunsaturated fatty acids contain multiple double bonds. Health aspects of a particular oil or oil mixture are generally believed to be dependant in part upon the fatty acid composition thereof.

In addition to being a useful source of energy, seed and nut derived oils provide nutrients such as essential fatty acids that are unavailable from other sources. Oils which have high saturated fatty acid compositions are not generally considered healthy oils as saturated fatty acids have been implicated in raising the level of total serum cholesterol, particularly the LDL component. While it is believed that oils such as palm oil promote health benefits due to their high beta carotene content, a precursor to vitamin A, it has been shown to demonstrate the same negative effect on cholesterol levels. Recent findings support use of oils having higher content of monounsaturated fatty acids to maintain serum cholesterol levels. Over the past several years, new varieties of traditionally available edible oils which have enhanced high monounsaturated fatty acid content have been created by natural selection plant breeding. Some of these varieties are High oleic Sunflower oil, High oleic safflower oil and High oleic Rape seed oil.

Another major aspect associated with the consumption of oil, is degradation. An oil is said to be stable when it demonstrates delayed lipid oxidation, can be stored for prolonged periods without trace of auto-oxidation. A stable oil will have the ability to be heated for long periods as well, such as when used for frying, before experiencing lipid oxidation under thermal stress. Although it is an important concern for all edible oils, oxidation degradation is of particular interest for oils used in frying. Thus, a higher stability is generally required for applications where oil is used for frying, be it in domestic, household use or in commercial use, such as in restaurants.

Vulnerability of various oils to degradation is affected by the fatty acid composition thereof. Generally, the higher the saturated fatty acid content, the more stable the oil would be. When raised to elevated temperatures, the rate at which the oil will degrade is greatly accelerated, and depending upon the fatty acid composition, the oil may be unacceptable for such use in its natural state. For example, of the traditional oil varieties mentioned above, only one or two of them have a fatty acid content which provides sufficient stability in the natural form to enable their use for professional frying or cooking.

Palm oil, although stable because of its high saturated fatty acid content, is not considered to be a healthy oil in the diet. Groundnut oil, also known as peanut oil, which has about 55% monounsaturated fatty acids and 26% polyunsaturated fatty acids, is more stable than other unmodified oils. However, it is not significantly more stable to make it a very attractive oil for frying, unless for other reasons than stability, such as a preference for the flavour of this oil. Soybean, sunflower and cottonseed oil are oils high in polyunsaturated fatty acids, and are therefore too unstable for prolonged use at elevated frying temperatures. Rape seed oil is also unacceptable, as it has an unusually high content of linolenic acid, approximately 10%, making this oil very unstable. In order to use the polyunsaturated fatty acid oils for cooking, frying and baking, they are chemically modified by hydrogenation. The triglyceride molecules that make up oils can be degraded either by reaction with water, called hydrolytic rancidity, or by reaction with oxygen, called oxidative rancidity. Rancidity due to hydrolysis can cause disagreeable odours and flavours, whereas oxidative rancidity is generally cited in the literature as a potential health concern. Such breakdown results in the reduction of nutritional value including the destruction of vitamins, as well as the formation of a number of potentially harmful byproducts, such as oxidized fatty acids, ketones, aldehydes, and possibly a number of mutagenic substances.

Oils other than saturated oils which are generally avoided due to their negative health implications, do not possess sufficient stability in their unmodified form for use in professional frying or other applications where thermal stress over prolonged periods is involved. Addition of synthetic anti-oxidants is also either ineffective because of their volatility or prohibition of commercial use by law. Although effective for frying foods and still extensively used, palm oil and the other so-called "tropical oils" have, in recent years, fallen out of favor in several countries due to sustained public campaigns implicating them as leading contributors to the development of atherosclerosis. These frying oils that are generally used tend to be originally unsaturated oils used in a partially hydrogenated modified form. It may be noted that during frying, the oil is invariably oxidized to some extent.

It is believed that there are very few frying oils in the market which can be regarded as healthy dietary components as they are either naturally saturated or partially hydrogenated. Thus, the introduction of hybrids of oil seeds having significantly higher monounsaturated fatty acid content is believed to provide a substantial health advantage. Due to the high proportion of monounsaturated fatty acids, the oil not only provides the purported health benefits of a cis-configured monounsaturated fatty acid, but is more stable than other unsaturated oils in their unmodified state. Some of these new varieties of traditionally used vegetable oils, such as the High Oleic Sunflower Oil (HOSO), High oleic Safflower Oil (HOSFO) and High Oleic Rape Seed (Canola) Oil, have less than 10% saturated fatty acids with 70% to 90% monounsaturated fatty acids. The most advanced variety currently in commercial use is HOSO manufactured in the United States (US4,627,192 and US4,743,402). The high monounsaturated fatty acid content in this oil is thought to reduce serum LDL.

It is imperative to increase the stability of an oil both in terms of storage and ensuring low oil uptake during cooking so that the essential consumption of oil is reduced to required levels for human beings. There are several studies which disclose increase in the stability of an oil for purposes of prolonged storage by the addition of a number of known synthetic anti-oxidants, such as butylated hydroxy anisole, butylated hydroxy toluene, propyl gallate, tertiary butyl hydroxyquinone; acid synergists, such as citric acid and its esters, monoglyceride citrate, ascorbic acid and its esters, ascorbyl palmitate; and foam inhibitors, such as methyl silicone, for effective delaying of lipid oxidation. However, where the oil is to be used at elevated temperatures for frying or baking, these substances tend to evaporate, distill or degrade rapidly under actual conditions of use, reducing the stabilizing effects thereof dramatically. Dimethyl polysiloxane, also referred to as methyl silicone, is used in many or most frying oils and is believed to act as a foam inhibitor.

US5,260,077 discloses an attempt to increase the stability of a high oleic oil by adding an anti-oxidant tocopherol in an effective amount. Studies however suggest that content of tocopherol in oil during frying depletes as much as 50% in just two hours under actual frying conditions at a temperature of 190°C. It is noted that all the experiments to which the cited patent application refers, were made employing Active Oxygen Method (A.O.M.), which indicates heating to 98°C.

It is also imperative to reduce the level of absorption of the oils into the food matrix in view of their implication in various diseases such as coronary heart diseases and obesity. Additionally, the storage of oils in the domestic or even commercial environments inevitably leads to a loss of stability due to thermal degradation. All edible oils are susceptible to oxidative degradation and result in the formation of primary and secondary oxidative products that adversely affect food quality. This phenomenon can occur at high thermal stress levels, whether applied voluntarily through the cooking process or during storage over periods of time. During deep fat frying, the fat is exposed to light, elevated temperature and atmospheric oxygen.

Another aspect of concern is the calorie content of the fried product. As consumers prefer reduced-calorie or light foods, there remains a need to provide such foods. Due to their excellent heat transfer properties, animal and vegetable fats and oils perform well as a cooking medium. Those familiar with the art will know that methyl siloxane among other silicone oils has been used to prepare low calorie foods either by partially or wholly replacing vegetable or animal oils and fats. However, this is associated with undesirable parameters such as excessive foaming and presence of typically developed fried flavours.

The basic requirements for fat and oil used for frying purposes are heat stability and oxidation stability. Additional requirements are that it should be cost-effective, have acceptable taste, be processable (e.g., flowable at ambient temperature) and organoleptically acceptable. Several attempts have been made in the art with varying degrees of success to provide edible oil compositions with enhanced stability. Some of the references are summarized below.

EP0326829 discloses the use of an anti-oxidant mixture in order to protect a fat from oxidation. The typical uses are in foodstuffs or cosmetic products that are sensitive to oxidation. The anti-oxidants used are tocopherol, ascorbic acid and lecithin. This mixture is claimed to be effective in protecting oils and fats that are rich in polyunsaturated fatty acids.

EP0577305 discloses a method for stabilizing polyunsaturates by adding ascorbic acid or an ester or salt thereof, and a phosphorylated mono- or di-fatty acyl glyceride or salt thereof, and optionally tocopherol or tocotrienol anti-oxidant. The primary uses disclosed are in the preparation of stable pharmaceutical, nutritional or veterinary compositions.

US4,765,927 discloses an anti-oxidant composition which includes tocopherol, BHT or BHA, condensed phosphates such as pentapolyphosphate and citric acid with good compatibility with fat, oil, and foods containing fat or oil.

US5,077,069 discloses a composition comprising natural anti-oxidants such as tocopherol, ascorbic acid, citric acid and phospholipids which are useful for the delayed oxidation of oils.

US20060110513 discloses a modified/improved frying process to enhance the stability of the oilduring frying.

WO2011/119462 discloses a stabilized frying oil containing one or more monovalent carbonate salts.

While more attention has been paid to enhancement of stability during storage and during frying, very little attention has been paid to maintain stability of the oil while in storage, in use in frying or other cooking methods where thermal stresses are induced or exist, and in vivo after consumption. The applicants herein adopted a holistic approach wherein it was realized that not only is it critical to reduce oxidative degradation during storage and in use, but is also critical to ensure that this phenomenon does not occur or is reduced after consumption of the food product cooked in the edible oil. The applicants are unaware of any attempts wherein a combined approach of enhancing stability, while reducing the absorption of the oil into the food matrix, without loss of functional parameters such as flavour, colour, taste and mouthfeel and ensuring that oxidative degeneration in vivo are avoided or minimized, has been employed.

OBJECT OF THE PRESENT INVENTION
It is an object of the invention to provide a premix for addition to an edible oil composition for enhanced stability during storage, in use in frying or other cooking methods, and in vivo after consumption of the food matrix, without deterioration in quality.

Another object of this invention is to provide a process for manufacturing a premix for addition to an edible oil composition and PUFA (poly unsaturated fatty acids) rich oils, which significantly increases the life of the oil.

Still another object of this invention is to provide a process for manufacturing a premix for addition to an edible oil composition which can be used for the preparation of deeply fried foodstuff with minimized retention of frying medium.

Still yet another object of this invention is to provide a premix for addition to an edible oil composition, comprising ascorbyl palmitate, tocopherol(s), tertiary butyl hydroquinone (TBHQ) and DMPS (dimethyl polysiloxane).

SUMMARY OF THE INVENTION
The present invention provides a premix for addition to an edible oil composition for enhanced stability during storage, in use in frying or other cooking methods, and in vivo after consumption of the food matrix. The premix of the present invention comprises a combination of anti-oxidants, which provide enhanced stability during shelf life, frying and in vivo. The embodiments of the present invention are not limited to cooking/edible oil for domestic household use and extend to all situations including the commercial use of cooking oil.

BRIEF DESCRIPTION OF DRAWINGS
Fig. 1 is a graphical representation of oil stability studies based on shelf life using the Rancimat index.
Fig. 2 is a graphical representation of studies carried out to measure peroxide value rise in the oils of the invention when compared with reference oils after frying.
Fig. 3 is a graphical representation of studies carried out to measure pAV (para anisidine value).
Fig. 4 is a graphical representation of studies carried out to measure the Totox combination.
Fig. 5 graphical representation of Rancimat™ studies carried out after repeated frying intervals conducted on 0th day till 4th day.
Fig. 6 graphical representation of low oil uptake studies carried out during frying studies.
Fig. 7 is a graphical representation of values obtained after animal model testing to determine the in vivo stability of the oil formulations of the invention containing a three anti-oxidant combination compared with a reference formulation which contains only TBHQ.
Fig. 8 is a graphical representation of in vivo studies to analyse the reduction of high sensitivity C-reactive protein (hsCRP) which is an inflammatory biomarker for the oil samples of the invention containing three anti-oxidants and reference oils which contain only TBHQ.

DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a premix for addition to an edible oil composition, comprising a combination of anti-oxidants that renders enhanced stability during storage, in use in frying or other cooking methods, and in vivo after consumption of the food matrix.

According to this invention, the premix for addition to the edible oil composition comprises: a) ascorbyl palmitate, b) tertiary butyl hydroquinone, c) tocopherol(s) and d) dimethyl polysiloxane.

The edible oil composition may comprise anti-oxidants selected from the group comprising of BHA, BHT, Lecithin, Gallets (propyl, octyl, etc), or their combinations.

According to the present invention, the composition comprises ascorbyl palmitate in an amount ranging from 50-300 ppm, preferably ranging from 200-300 ppm.

The composition of the present invention comprises natural or synthetic tocopherols in an amount ranging from 300-1000 ppm.

According to the present invention, the composition comprises TBHQ in an amount ranging from 100-200 ppm.

According to the present invention, the composition comprises DMPS in an amount ranging from 1-10 ppm.

The edible oil or oils in combination may additionally comprise one or more food grade additives such as vitamins, minerals or nutrients.
The synergistic premix of the present invention is obtained by mixing ascorbyl palmitate, mixed tocopherols, TBHQ and DMPS in an oil base, stirring and heating under stirring to a temperature in the range of 80 to 125°C. The mixture is stirred till homogeneity is obtained. This premix is added to the edible oil or combination of edible oils in a measured dosage form till the entire premix is used. The edible oil composition so obtained is further stirred or recirculated for two to four hours till homogeneity is obtained. Additional additives such as vitamins, minerals and other food grade additives may be added if desired in proportions that are conventional. The oil soluble vitamins can be uniformly mixed in the oil using a stirrer. The addition of the anti-oxidant composition and oil/fat soluble vitamins not only increases the lower uptake of oil by the fried food stuff, but also results in retaining the higher nutrients in the oil and higher sensory values such as colour, aroma and mouthfeel in the food stuff prepared. This is essentially due to the synergy achieved by adding the anti-oxidant combination to the edible oil.

In a preferred form of this invention, the cooking medium is edible oil or fat or a combination of edible oils and fats typically used in frying. This invention does not claim to be the broad solution to the general problem of rancidity due to oxidation, but rather to an improved and relatively simple solution to that problem.

It is also recognized that the cost of the cooking medium is also a key factor in any commercial, or semi-commercial operation, e.g. school lunch programs, in which a significant portion of the products prepared are deep fat fried. Hydrogenated vegetable oils and some of the rarer cooking oils are quite stable to oxidation, but may be subject to flavour reversion. The tendency, however, seems to be to use the least expensive product and to use that product for as long as it can be used, consistent with the quality criteria adopted by the user. In the case of oils containing substantial amounts of linoleic acids, for instance, soybean oil or rapeseed oil, the oxidation is generally referred to as "flavour reversion" and generally requires a lower degree of oxidation than may be required for oxidative rancidity. These oils, however, may be less expensive and may be the oils of choice or may be blended with higher priced oils in order to increase the useful life of the cooking medium. The stability of a cooking medium is usually expressed in terms of AOM, which is an indication of the rancidity point of the cooking medium. It is generally recognized that when the peroxide content of the cooking medium reaches 70 meq/kg of oil, the rancidity point has been reached. It is also recognized that once the rancidity point has been reached, the addition of anti-oxidant material is not effective to reverse the process. It thus becomes significant to control or inhibit the formation of the products contributing to rancidity of the cooking medium and the resulting products. In accordance with this invention, the food-grade and accepted anti-oxidant materials are used in an effective and permissible amount and in an automatically controlled application to provide the optimum control as permitted by the prevailing safety regulations. The anti-oxidant material may be any of the natural products mentioned or those known in the art, and which are acceptable for use, alone or in combination. Most anti-oxidants interfere with the propagation of lipid oxidation by donating a hydrogen atom to, and thereby inactivating, chain-carrying peroxyl, radicals and/or alkoxyl radicals. Hence, after breaking the chain reaction of lipid peroxidation, an anti-oxidant is itself converted to a radical. To be effective, the anti-oxidant radical has to be sufficiently stable so as to react slowly with the lipid substrate and rapidly with lipid peroxyl radicals and/or alkoxyl radicals. Several natural and synthetic compounds fulfil this condition and are widely used for preserving polyunsaturated fatty acids (PUFA) from oxidative deterioration.

Auto-oxidation of highly unsaturated fatty acids can yield hard, tough, insoluble polymeric products. In the final stage of glyceride oxidation, hydroperoxides split or decompose into short-chain aromatic compounds (mainly aldehydes, ketones, alcohols, and acids), which are responsible for the rancidity condition that ultimately destroys acceptability and usefulness of fats and oils. Auto-oxidation is initiated or strongly catalyzed by a number of factors. Heat, for instance, greatly accelerates oxidation, especially at higher temperatures (above 60°C) where it has been estimated that for each 10°C increase in temperature, the rate of oxidation reaction doubles. Another important factor is the presence of metals, which in just trace amounts are recognized as the predominant pro-oxidant materials encountered in commercial fats and oils. It is estimated that copper or iron at concentrations of less than 1 ppm can cause very serious reduction of fat and oil stability. This problem is magnified by free fatty acids which act to solubilize metals in fats or oils. As the glyceride auto-oxidation is initiated and promulgated by the formation of free radicals, the removal or inactivation of the fatty or peroxide free radicals should terminate, or at least interrupt, fat oxidation in its early stages, and thus delay breakdown into the final end products that are responsible for rancidity.

TBHQ is added at a concentration preferably in the range of 100 – 200 ppm to suppress the development of peroxides, critical particularly during food storage. The anti-oxidative activities of these synthetic substances stem from the phenolic configuration of their molecular structures. These phenolic substances function as free radical acceptors which can terminate fat or oil oxidation at the initiation stage. The anti-oxidant free radical that forms is stable and, most importantly, does not initiate or promulgate further oxidation of the glyceride.

Ascorbyl palmitate is an ester formed from ascorbic acid and palmitic acid. In addition to its use as a source of vitamin C, it is also commonly used as an anti-oxidant food additive. Unlike the primary anti-oxidants which function as electron donors, ascorbyl palmitate functions by oxygen scavenging. Ascorbyl palmitate is used in an amount of 50 to 300 ppm, preferably 200-300 ppm. Ascorbyl palmitate can be weighed directly into oil premix and added to the oils. Solubility of 0.05 percent in oils can be achieved.

Tocopherols are among the most important lipid-soluble natural anti-oxidants, and appear to be the major physiological scavengers of free radicals inside human membranes and plasma lipids. The fact that these compounds are naturally occurring lipid-soluble anti-oxidants make them particularly useful in combination with oils having high amounts of PUFA, intended for human consumption. It has long been recognized that tocopherol molecules with the phenolic configuration exhibit anti-oxidant properties. There are at least four types of tocopherol with α-, γ-, and δ- isomers predominating in vegetable matter. Vitamin E activity is attributed mainly to α-tocopherol, which also provides some oxidation inhibition effect in oil, but the γ- and δ- forms are more effective anti-oxidants. Tocopherols are widely distributed in many vegetable matters from which commercial edible oils are extracted. The tocopherols used in the present invention may be natural or synthetic or a combination thereof and preferably used in 300-1000 ppm.

The following examples are illustrative of the practice of the invention, but the invention is not limited to the specific illustrations provided. Separate batches of pre-mixes of the anti-oxidant combination were prepared by mixing with a base oil and heating to a temperature in the range of 80 - 125°C. The mixture was heated and stirred under heating till homogeneity was obtained. Any conventional homogenizer can be used for the preparation of the premix. Thus obtained premix is added in separate batches to edible oils under stirring using inline dosing till a homogeneous mixture is obtained. To this homogeneous edible oil mixture, desired oil soluble vitamins and minerals may be added to enhance nutritive value. In the illustrative examples, “reference oils” represent oils that contain only TBHQ which were considered for comparison of properties with the oils of the invention which comprise a combination of three anti-oxidants.

Fig. 1 is a graphical representation of oil stability studies (shelf life) using the Rancimat index. The oil stability index results were measured in terms of the induction hours using an instrument called Rancimat™ at 110°C. Higher stability of the oil is demonstrated by higher induction times. As can be seen from Fig. 1, the formulation of the invention comprising rice bran oil and safflower oil blend comprising the anti-oxidant combination (RBO+SAFF+3A) and high oleic safflower oil comprising anti-oxidant combination (HOSFO + 3A) showed higher induction times compared to the reference oils.

Fig. 2 is a graphical representation of studies carried out to measure peroxide value rise in the oils of the invention in comparison with reference oils, after frying. PV (peroxide value) is the primary oxidation product which gets generated in the oil during frying. As can be seen from Fig. 2, the peroxide values of the oil formulations of the invention are significantly less compared to the reference oils.

Fig. 3 is a graphical representation of studies carried out to measure p-AV (para-anisidine value). p-AV is a secondary oxidation product that is generated in oil during frying. As can be seen from Fig. 3, the oil formulations of the invention show a lower rise in p-AV compared to reference oils.

Fig. 4 is a graphical representation of studies carried out to measure the Totox (total oxidation products (2*PV+p-AV) combination). It is the extent of generation of the oxidation product during frying. As is evident from Fig. 4, the oil formulations of the invention show a lesser rise in Totox values compared to the reference oils.

Fig. 5 are graphical representation of Rancimat™ studies carried out after repeated frying. The oils of the present invention show higher induction time compared to that by the reference oils.

Fig. 6 is graphical representation of low oil uptake studies carried out during frying. The oils of the present invention show lesser oil uptake as compared to that by the reference oils.

Fig. 7 is a graphical representation of values obtained after animal model testing to determine the in vivo stability of the oil formulations of the invention in comparison with a reference control animal group. The values depicted are for the levels of the three enzymes catalase, super oxide dismutase and glutathione in Wistar rats.
The animal model testing was carried out to determine the stability of the oil formulations of the present invention and control animal group, and to determine the effect of these oil blends on free radical generation. As is evident from Fig. 7, the levels of the three enzymes tested in the animal models were higher in the case of the oil samples of the invention compared to control animal group. This clearly demonstrates that the oil formulations of the invention lead to the inhibition of free-radical generation.

Fig. 8 is a graphical representation of in vivo studies to analyse the reduction of high sensitivity C-reactive protein (hsCRP) which is an inflammatory biomarker with the oil samples of the invention in comparison with control animal group. Relatively high levels of hs-CRP in otherwise healthy individuals have been found to be predictive of an increased risk and potential future heart attack, stroke, sudden cardiac death and/or peripheral arterial disease, even when cholesterol levels are within an acceptable range. As is evident from Fig. 8, the oil samples of the invention demonstrated reduced levels of hs-CRP, compared to the control animal group, thereby demonstrating enhanced in vivo stability thereof.

EXPERIMENTAL EXAMPLES

EXAMPLE 1
A batch of 2000 gm of oil formulation is made by preparing a premix comprising of 0.3 gm of tertiary butyl hydroquinone, 0.54 gm of ascorbyl palmitate (20% product in soya oil base), 0.84.gm of mixed tocopherols, and 0.016 gm of DMPS with 12 gm of the oil base, using a mixer at 1000 rpm for a period of 30 min while maintaining the temperature in the range of 80 - 125°C. The premix is then added to a portion of pure high oleic safflower oil. The premix doses obtained above are added under heating and stirring to form a homogeneous mixture. The mixer is operated at 200 rpm for a period of 30 min while maintaining room temperature. The proportions of the oil composition so obtained are presented in Table 1.

Table 1:
Constituent of the oil Quantity (gm)
High Oleic Safflower Oil 1998.30
Ascorbyl palmitate 0.54
Mixed tocopherols 0.84
TBHQ 0.3
DMPS 0.016
EXAMPLE 2
A batch of 2000 gm of oils formulation is prepared by preparing a premix comprising of 0.3 gm of tertiary butyl hydroquinone, 0.54 gm of ascorbyl palmitate (20% product in soya oil base), 0.84gm of mixed tocopherols, and 0.016 gm of DMPS with12 gm of the oil base and using a mixer at 1000 rpm for a period of 30 min while maintaining the temperature in the range between 80 - 125°C. The premix is then added to a portion of a blend of Rice bran oil and Linoleic safflower seed oil. The premix doses obtained above are added under heating and stirring to form a homogeneous mixture. The mixer is operated at 200 rpm for a period of 30 min while maintaining the temperature at room temperature. The proportions of the oil composition so obtained are presented in Table 2.
Table 2:
Constituent of the oil Quantity (gm)
Linoleic Safflower oil +
Rice bran oil (30:70) 1998.30
Ascorbyl palmitate 0.54
Mixed tocopherols 0.84
TBHQ 0.3
DMPS 0.016

For the comparative/ efficacy studies, oils of the present invention (Examples 1 and 2) comprise the combination of the anti-oxidants while the Reference oils considered are soybean, sunflower, groundnut, olive (extra virgin and pomace), canola, rice bran (market samples consisting of only one antioxidant, viz., TBHQ). The market samples were picked up randomly from grocery store.

EXAMPLE 3
Bread croutons were fried in reference oil and the said oils of examples 1 and 2, maintaining the same frying conditions. The food items fried in the said oil compositions of the invention showed lower oil uptake. The data is provided in Fig. 6. As is evident, the oil uptake by fried food is lesser for oil compositions of the invention when compared with those of the reference oils.

EXAMPLE 4:
Extended shelf life stability studies were carried out using RANCIMAT OSI with the reference oil and the oils obtained by examples 1 and 2 above. As is evident from Fig. 1, the oils of examples 1 and 2 displayed a significant increase in shelf life stability over reference oils.
EXAMPLE 5:
A High Oleic Safflower Oil (HOSO) composition comprising 270 mg/kg of ascorbyl palmitate, 150 mg/kg of TBHQ, 420 mg/kg of mixed tocopherols and 8 mg/kg of DMPS, was prepared following the procedure of Examples 1 and 2 detailed above. Various food items were fried in these oil compositions as well as the reference oil while maintaining similar frying conditions. The food items were bread cubes. These items cooked quickly, had an aesthetically pleasing appearance, did not burn and were not excessively "greasy" when cooked in the oil of the present invention. The taste was also excellent in each case, compared to the reference oil fried foodstuff.

Bread cubes were fried in oil compositions prepared as described in example 1 and 2 and in the above reference oil. In comparison with the reference oil, oil uptake in bread cubes was significantly low in the said oil compositions of the present invention.

Frying performance of the oil was analysed by the chemical parameters such as peroxide value, anisidine value and Totox value, rancimat (data is provided in fig 2, fig 3, fig 4 & fig 5) and compared with the reference oils (i.e. market samples) which in the case of the frying studies are soybean, sunflower, groundnut, olive (extra virgin and pomace), canola, rice bran (market samples consisting of only antioxidant i.e. TBHQ). The market samples were picked up randomly from grocery store. As is evident, the peroxide value, the anisidine value and the total oxidation value of the oils obtained for oil compositions of the invention are far superior to those of the reference oils.

For in vivo benefit, various parameters such as changes in Lipid profile (LDL level, Total cholesterol, HDL level, VLDL level, Triglycerides), Cardiac markers (hsCRP, ox LDL, Lipoprotein (a), Apolipoprotein A1, Apolipoprotein B, Homocysteine, ratio of Apolipoprotein B to Apolipoprotein A1) were also studied. The oils of the present invention have been subsequently represented as “Test group” while the oils comprising only TBHQ are represented as “Reference group”, in the below described evaluation and comparative studies. Reference oils are market samples for sunflower oil, corn oil, coconut oil, palm oil & ghee. The market samples were picked up randomly from grocery store.

LIPID PROFILE
Comparison of changes in mean LDL level (mg/dl):
Fig. 5 presents the comparative study of the changes in the mean LDL level of the Test group and Reference oils. The study reveals that the mean LDL Level was 155.17 mg/dl among the Reference group which was more though not significant, compared to 160.53 mg/dl of the Test group at baseline. At day 30, mean LDL Level showed a significant fall of 16.7% among Reference group and 27.0% in Test group from baseline. After 60 days of treatment, the decline in the mean LDL Level of the Reference group was 19.3% while it was significant with a decline of 32.0% in the case of Test group. At day 90, the decline in the mean LDL Level was a significant 20.6% in the case of the Reference group and 34.9% in the case of the Test group from baseline. The results are presented below in table 3.
TABLE 3
Duration (Days) Mean LDL Level (mg/dl) (  SD) p value
Reference (N=41) Test (N=39)
Baseline 155.17 + 13.04 160.53 + 17.45 0.125 (NS)
30 129.19 + 13.16 117.20 + 15.07
60 125.16 + 19.72 109.18 + 14.82
90 123.19 + 13.23 104.46 + 18.17
Mean Diff. (Baseline – Day 30)
(p value) *-25.98 + 05.46
(0.001) *-43.33 + 09.25
(0.001) *0.001
Mean Diff. (Baseline – Day 60)
(p value) *-30.01 + 16.57
(0.001) *-51.35 + 09.87
(0.001) *0.001
Mean Diff. (Baseline – Day 90)
(p value) *-31.98 + 03.81
(0.001) *-56.07 + 04.31
(0.001) *0.001
By ANOVA N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean total cholesterol (mg/dl):
The changes in the mean Total Cholesterol of the Test group were assessed and compared with that of the Reference group. Fig. 6 represents the comparative study results. The study suggests that the mean Total Cholesterol was 232.47 mg/dl among the Reference group which was more though not significant, compared to 236.92 mg/dl of the Test group at baseline. At day 30, the decline in the mean Total Cholesterol was 12.3% in case of the Reference group, which was more though not significant, compared to 18.7% of the Test group from baseline. At day 60, the decline in the mean Total Cholesterol was 13.0% in the case of the reference group, which was more though not significant, compared to 20.6% of the Test group from baseline. At the end of the treatment, the decline in the mean Total Cholesterol was 13.8% in the case of the reference group, which was again more though not significant, compared to 22.1% of the Test group from baseline. The study results are presented below in table 4.
TABLE 4
Duration (Days) Mean Total Cholesterol (mg/dl) (  SD) p value
Reference (N=41) Test (N=39)
Baseline 232.47 + 13.34 236.92 + 10.55 0.101 (NS)
30 203.94 + 15.17 192.54 + 17.80
60 202.19 + 14.07 188.07 + 15.95
90 200.49 + 13.03 184.61 + 17.82
Mean Diff. (Baseline – Day 30)
(p value) *-28.53 + 06.68
(0.001) *-44.38 + 13.64
(0.001) *0.001
Mean Diff. (Baseline – Day 60)
(p value) *-30.28 + 07.14
(0.001) *-48.85 + 11.43
(0.001) *0.001
Mean Diff. (Baseline – Day 90)
(p value) *-31.98 + 04.12
(0.001) *-52.31 + 13.04
(0.001) *0.001
By ANOVA NS = Not Significant *Significant

Comparison of changes in mean HDL level (mg/dl)
The changes in the mean HDL level of the Test group were assessed and compared with that of the Reference group. Fig. 7 represents the comparative study results. As per this analysis, the mean HDL level was 38.47 mg/dl among the Reference group and 39.76 mg/dl among the Test group at baseline, the difference being not statistically significant. After 30 days, the increase in mean HDL level was 6.5% in case of Reference group while the increase was 12.9% in case of Test group. It was observed that the increase in mean HDL level was more among Test group than Reference group and the difference was statistically significant. At day 60, the increase of mean HDL level was 27.8% in case of the Test group, which was significantly more than the increase of 12.1% in case of the Reference group. At day 90 of treatment, the mean HDL level showed a significant increase of 17.2% and 32.7% among Reference and Test group respectively. When compared the increase was significantly more among Test group than Reference group. The results are presented in table 5 below.
TABLE 5
Duration (Days) Mean HDL Level (mg/dl)(  SD) p value
Reference (N=41) Test (N=39)
Baseline 38.47 + 06.88 39.76 + 06.53 0.392 (NS)
30 40.96 + 06.42 44.90 + 06.43
60 43.12 + 06.29 50.80 + 06.95
90 45.07 + 07.21 52.75 + 06.28
Mean Diff. (Baseline – Day 30)
(p value) *02.49 + 02.26
(0.001) *05.14 + 01.95
(0.001) *0.001
Mean Diff. (Baseline – Day 60)
(p value) *04.65 + 02.81
(0.001) *11.04 + 03.08
(0.001) *0.001
Mean Diff. (Baseline – Day 90)
(p value) *06.60 + 01.40
(0.001) *12.99 + 01.00
(0.001) *0.001
By ANOVA N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean VLDL level (mg/dl)
The changes in the mean VLDL level of the Test group were assessed and compared with that of the Reference oils. Fig. 8 presents the comparative study results. The study states that, the mean VLDL level was 38.88 mg/dl among Reference group which was comparable with 38.61 mg/dl among the Test group at baseline but the difference was not statistically significant. At day 30, the decline in mean VLDL level was 13.1% in case of Reference group while the decline was 23.2% in case of Test group. It was observed that the fall in mean VLDL level was more among Test group than Reference group and the difference was statistically significant. After 60 days of treatment, the decline in mean VLDL level was 27.3% in case of the Test group, which was significantly more than the fall of 14.4% in case of the Reference group. At day 90, the decline in the mean VLDL level was significant 17.1% in the case of the Reference group and 29.0% in the case of the Test group from baseline. It was observed that the decline was more among Test group then Reference group and the difference was statistically significant. The study results have been summarised in table 6 below.

TABLE 6
Duration (Days) Mean VLDL Level (mg/dl)(  SD) p value
Reference (N=41) Test (N=39)
Baseline 38.88 + 10.53 38.61 + 08.61 0.900 (NS)
30 33.80 + 10.56 29.66 + 07.70
60 33.29 + 10.21 28.08 + 07.50
90 32.23 + 10.80 27.40 + 08.47
Mean Diff. (Baseline – Day 30)
(p value) *-05.08 + 04.45
(0.001) *-08.95 + 02.52
(0.001) *0.001
Mean Diff. (Baseline – Day 60)
(p value) *-05.59 + 04.27
(0.001) *-10.53 + 03.25
(0.001) *0.001
Mean Diff. (Baseline – Day 90)
(p value) *-06.65 + 01.19
(0.001) *-11.21 + 00.69
(0.001) *0.001
By ANOVA N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean triglycerides (mg/dl)
The changes in the mean Triglycerides level of the Test group were assessed and compared with that of the Reference oils. Fig. 9 presents the comparative study results. From the below data, the mean Triglycerides level was 194.40 mg/dl among the Reference group which was comparable with 193.05 mg/dl among the Test group at baseline but the difference was not statistically significant. After 30 days of treatment, the decline in the mean Triglycerides was 12.6% in case of the Reference group while the decline was 24.1% in case of the Test group from baseline. It was observed that the fall in the mean Triglycerides was more among the Test group than the Reference group and the difference was statistically significant. At day 60, the decline in the mean Triglycerides was 27.3% in case of the Test group, which was significantly more than the fall of 13.1% in case of the Reference group. At the end of day 90, the decline in the mean Triglycerides was significant 17.1% in case of the Reference group and 29.0% in case of the Test group from baseline. It was observed that the decline was more among the Test group than the Reference group and the difference was statistically significant. The study results are represented below in Table 7.

TABLE 7
Duration (Days) Mean Triglycerides (mg/dl) (  SD) p value
Reference (N=41) Test (N=39)
Baseline 194.40 + 52.65 193.05 + 43.07 0.900 (NS)
30 169.93 + 52.33 146.56 + 37.62
60 168.92 + 51.86 140.39 + 37.52
90 161.14 + 53.98 137.02 + 42.37
Mean Diff. (Baseline – Day 30)
(p value) *-24.47 + 21.89
(0.001) *-46.49 + 14.27
(0.001) *0.001
Mean Diff. (Baseline – Day 60)
(p value) *-25.48 + 23.92
(0.001) *-52.66 + 16.23
(0.001) *0.001
Mean Diff. (Baseline – Day 90)
(p value) *-33.26 + 05.93
(0.001) *-56.03 + 03.43
(0.001) *0.001
By ANOVA N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

CARDIAC MARKERS
Comparison of changes in mean hsCRP
The changes in the mean hsCRP of the Test group were assessed and compared with that of the Reference oils. Fig. 10 presents the comparative study results. This profile reveals that the mean hsCRP was 2.85 among the Reference group which was comparable to 3.61 among the Test group at baseline and thus the difference was not statistically significant. At day 90, the mean hsCRP showed an insignificant rise of 15.4% among the Reference group and showed an insignificant fall of 24.1% in the Test group from baseline. It was observed that the change was more among the Test group than the Reference group and the difference was statistically significant. This result reveals that the mean hsCRP among the Test group had more than 2 times fall as compared to the Reference group. The study results are summarised below in table 8.

TABLE 8
Duration (in Days) Mean hsCRP (mg/L)(  SD) p value
Reference (N=40) Test (N=34)
Baseline 2.85 + 3.04 3.61 + 3.83 0.383 (NS)
90 3.29 + 3.90 2.74 + 2.38
Mean Diff. (Baseline – Day 90)
(p value) One-tailed 0.44 + 2.37
(0.124) NS -0.87 + 2.85
(0.957) NS
*0.018
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean ox LDL
The changes in the mean ox LDL of the Test group were assessed and compared with that of the Reference oils. Fig. 11 presents the comparative study results. This data suggests that the mean ox LDL was 122.49 among the Reference group which was comparable to 123.91 among the Test group at baseline and hence the difference was insignificant statistically. After treatment at Day 90, the mean ox LDL showed a significant rise of 0.8% among the Reference group and a significant fall of 1.8% in the Test group from baseline. It was observed that the change was significantly more among the Test group than the Reference group.The study results are summarised below in table 9.
TABLE 9
Duration (in Days) Mean ox LDL (  SD) p value
Reference(N=41) Test (N=39)
Baseline 122.49 + 32.58 123.91 + 29.94 0.839 (NS)
90 123.53 + 32.71 121.68 + 29.78
Mean Diff. (Baseline – Day 90)
(p value) One-tailed
(p value) Two-tailed *1.04 + 1.73
(0.001)
(0.001) *-2.23 + 1.30
(0.001)
(0.001)
*0.001
*0.001
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean Lipoprotein (a)
The changes in the mean Lipoprotein (a) of the Test group were assessed and compared with that of the Reference oils. Fig. 12 presents the comparative study results. The analysis states that the mean Lipoprotein (a) was 22.66 mg/dl among Reference group which was more as compared to 16.87mg/dl among the Test group at baseline but the difference was not significant statistically. At the end of Day 90, the mean Lipoprotein (a) showed insignificant rise of 0.8% among Reference group and showed insignificant fall of 1.0% in Test group from baseline. It was observed that the change was comparable between the groups and difference was insignificant. The results of the analysis are shown below in table 10.
TABLE 10
Duration (in Days) Mean Lipoprotein (a) (mg/dl)(  SD) p value
Reference (N=41) Test (N=39)
Baseline 22.66 + 20.96 16.87 + 16.71 0.175 (NS)
90 22.85 + 21.85 16.70 + 15.39
Mean Diff. (Baseline – Day 90)
(p value) 0.19 + 4.76
(0.799) -0.17 + 7.98
(0.895)
0.808 (NS)
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant

Comparison of changes in mean Apolipoprotein A1
The changes in the mean Apolipoprotein A1 of the Test group were assessed and compared with that of the Reference oils. Fig. 13 presents the comparative study results. According to this study the mean Apolipoprotein A1 was 120.38 mg/dl among the Reference group which was comparable to 120.01 mg/dl among the Test group at baseline and thus the difference was statistically insignificant. After treatment at day 90, the mean Apolipoprotein A1 showed insignificant fall among both the groups i.e. 3.2% among the Reference group and 1.6% in the Test group from baseline. It was observed that the change was comparable between the groups and the difference was not significant. The study results are summarised below in table 11.
TABLE 11
Duration (in Days) Mean Apolipoprotein A1 (mg/dl)(  SD) p value
Reference(N=41) Test(N=39)
Baseline 120.38 + 23.87 120.01 + 17.76 0.937 (NS)
90 116.50 + 22.31 118.13 + 17.88
Mean Diff. (Baseline – Day 90)
(p value) -3.88 + 23.24
(0.291) -1.88 + 21.72
(0.592) 0.692 (NS)
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant

Comparison of changes in mean Apolipoprotein B
The changes in the mean Apolipoprotein B of the Test group were assessed and compared with that of the Reference oils. Fig. 14 presents the comparative study results. This profile states that the mean Apolipoprotein B was 110.54 mg/dl among the Reference group which was more as compared to 102.54 mg/dl among Test group at baseline however the difference was statistically not significant. At Day 90, the mean Apolipoprotein B showed significant fall of 16.4% among Reference group and 17.2% in the Test group from baseline. It was observed that the change was comparable between the groups and difference was insignificant. The study results are summarised below in table 12.
TABLE 12
Duration (in Days) Mean Apolipoprotein B (mg/dl)(  SD) p value
Reference(N=41) Test (N=39)
Baseline 110.54 + 28.74 102.54 + 34.92 0.268 (NS)
90 092.37 + 19.47 084.86 + 25.21
Mean Diff. (Baseline – Day 90)
(p value) *-18.17 + 30.70
(0.001) *-17.68 + 37.73
(0.006)
0.949 (NS)
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean homocysteine
The changes in the mean Homocysteine of the Test group were assessed and compared with that of the Reference oils. Fig. 15 presents the comparative study results. The study indicates that the mean Homocysteine was 18.46 among Reference group which was comparable to 19.24 among the Test group at baseline and hence the difference was insignificant statistically. After treatment at the end of Day 90, the mean Homocysteine showed insignificant fall of 2.0% among the Reference group and a significant fall of 11.2% in the Test group from baseline. It was observed that the change was more in the Test group than the Reference group and the difference was statistically significant. This result reveals that, the mean Homocysteine among the Test group had more than 2 times fall as compared to the Reference group. The study results are presented below in table 13.

TABLE 13
Duration (in Days) Mean Homocysteine (umol/L)(  SD) p value
Reference(N=40) Test (N=34)
Baseline 20.35 + 11.29 20.38 + 09.94 0.990 (NS)
90 19.94 + 10.57 18.09 + 09.49
Mean Diff. (Baseline – Day 90)
(p value) -0.41 + 03.76
(0.494) NS -2.29 + 3.97
*(0.002)
*0.041
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

Comparison of changes in mean ratio of Apolipoprotein B to Apolipoprotein A1
The changes in the mean ratio of Apolipoprotein B to Apolipoprotein A1 of the Test group were assessed and compared with that of the Reference oils. Fig. 16 presents the comparative study results. The study reveals that the mean Apolipoprotein B / A1was 0.93 among the Reference group which was comparable with 0.85 among the Test group at baseline and the difference was not significant. At day 90, the mean Apolipoprotein B / A1showed a significant fall of 11.8% among Reference group and 14.1% in Test group from baseline. It was observed that the fall was more among Test groups than Reference group and difference was not statistically significant. The study results are presented below in table 14.
TABLE 14
Duration (in Days) Mean Apolipoprotein B / A1(  SD) p value
Reference(N=41) Test (N=39)
Baseline 00.93 + 00.20 00.85 + 00.24 0.115(NS)
90 00.82 + 00.21 00.73 + 00.22
Mean Diff. (Baseline – Day 90)
(p value) -00.11 + 00.18
*(0.001) -00.12 + 00.24
*(0.002)
0.806 (NS)
By Student‘t’ Test N is number of subjects lasts till the end of study in each group; NS = Not Significant; *Significant

It must be understood that variations and modifications are possible without departing from the scope and spirit of the invention.