DESC:TECHNICAL FIELD
The present invention relates to a composition comprising extracts of green tea and rosemary extract optionally along with adjuvant and/or excipient. In addition, the present invention relates to a process for the preparation of said composition. The instant invention further relates to a composition comprising extract of green tea, rosemary extract and synthetic antioxidant optionally along with adjuvant and/or excipient.

BACKGROUND AND PRIOR ART OF THE INVENTION
Currently, food preservation systems often use chemicals and heat treatments to reduce the risk of bacterial food poisoning outbreaks and food spoilage, but chemicals can alter the taste of the product and, moreover, can compromise food safety.
In the past several years, a lot of work has been done in developing products that would serve as preservatives that could prevent the oxidation of fats, vegetable oils, carotenoids and their biologically active derivatives such as essential oils and other flavoring products so that the degradation of their quality is prevented in foodstuffs. Fats, oils and their products become rancid or unpleasant by odor or flavor due to the oxidative effects. The prior art shows various methods of inhibiting oxidation by addition of fat soluble antioxidants to the foodstuff. A number of chemical compounds are employed for avoiding or reducing these effects, so that, fats and oils or food containing fats and oils can be kept for longer periods of time. However, such agents have not been satisfactorily effective in many ways.
The chemical anti-oxidants that had been made use of in such cases include BHA (butylated hydroxyanisole), BHT (butylated hydroxyl toluene) and TBHQ (tertiary butyl hydroquinone) as well as other chemicals such as propyl gallate (PG). However, their volatility and tendency to decompose at higher temperature make them less suitable for deep fat or oil fried foods. Also, they were not found to be effective in protecting certain off-flavor development or the so called reversion flavor that occurs with the passage of time in oils like soybean oil.
It has been found that certain plant materials or extracts such as grape seed extracts, green tea extracts, sage, clove bud oil, clove leaf oil, Vitamin C, cinnamon leaf oil, oleoresin turmeric, tocopherol, tocotrienol, rosemary extracts and gallic acid etc., or salts thereof, were having anti-oxidant properties and their use as anti-oxidant preservatives have been widely discussed in past two decades. The use of natural anti-oxidants such as green tea as stabilizers for fats, oils, fatty food and ingredients of food is also discussed widely in US Patent No. 3812266 and US Patent No. 3451832. These patents describe the use of green tea and the importance of green tea catachin in the anti-oxidant activity of green tea. The work done in US Patent No. 4840966 and CA Patent No. 1057113A1 shows the health benefits of green tea and other such natural anti-oxidants.
Green tea is made from unfermented leaves and reportedly contains the highest concentration of powerful antioxidants called polyphenols, high antioxidant activity of green tea extracts, are used as a kind of innovative food additive to preserve pork, chicken meat, vegetable oil, fish oil and fish flesh, food emulsions and animal fat. Even though a number of antioxidants and various combinations thereof have been disclosed in the various inventions, there is still a need for additional antioxidant compositions in its right formulator form having improved characteristics.
With all the health benefits and the advantages of being all natural products, the use of such natural extracts needed further development of processes of extraction and the development of a right formulator form thereby making them effectively usable in such applications. The solubility factor is most important in such cases, where the formulated product had to be oil and fat soluble for them to be effective in their action as antioxidants.
In recent past several products are formulated for various application in fats and oil applications. Though the products developed from these natural products so far were found to be effective in their use as anti-oxidants, their use was not found satisfactory in terms of solubility factor with respect to time, sedimentation and extended shelf life in its use as preservatives in oils and fats and in processed foods such as deep fried foodstuffs. Thus, there was a need to develop a composition that can successfully overcome the difficulties stated above.
SUMMARY OF THE INVENTION
Accordingly the present disclosure relates to a composition comprising green tea extract and rosemary extract, optionally along with adjuvant or excipient or a combination thereof; a process of preparing the composition comprising green tea extract and rosemary extract, said process comprising acts of: a) granulating crude extract of green tea and mixing the granulated extract with the rosemary extract to obtain a mixture, b) optionally adding adjuvant or excipient or a combination thereof to the mixture, and c) passing the mixture of step a) or b) through homogenizer to obtain said composition comprising green tea extract and rosemary extract; a composition comprising the composition as mentioned above and tertiary butyl hydroquinone; an oil, fat or cosmetic preparation, comprising the composition as mentioned above; and a method of preparing the oil, fat or cosmetic preparation as mentioned above, wherein said method comprises act of mixing the composition as mentioned above with the oil, fat or cosmetic ingredients.

BRIEF DESCRIPTION OF THE ACCOMPANYING FIGURES
Figure 1A illustrates Oxidative stability of palm oil at temperatures 100ºC, 120 ºC and 140 ºC upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 1B illustrates increase in the protection factor of palm oil at 100ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 1C illustrates increase in the protection factor of palm oil at 120ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 1D illustrates increase in the protection factor of palm oil at 140ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 2A illustrates Oxidative stability of sun flower oil at temperatures 80ºC, 100 ºC and 120 ºC upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 2B illustrates increase in the protection factor of sun flower oil at 80ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 2C illustrates increase in the protection factor of sun flower oil at 100ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 2D illustrates increase in the protection factor of sun flower oil at 120ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 3A illustrates Oxidative stability of almond oil at temperatures 100ºC, 120 ºC and 140 ºC upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 3B illustrates increase in the protection factor of almond oil at 100ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 3C illustrates increase in the protection factor of almond oil at 120ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 3D illustrates increase in the protection factor of almond oil at 140ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 4A illustrates Oxidative stability of apricot oil at temperatures 100ºC, 120 ºC and 140 ºC upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 4B illustrates increase in the protection factor of apricot oil at 100ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 4C illustrates increase in the protection factor of apricot oil at 120ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 4D illustrates increase in the protection factor of apricot oil at 140ºC, upon addition of instant composition, sample B, sample B1 and sample B2, respectively.
Figure 5A illustrates Oxidative stability of fish oil at temperatures 60ºC, 70 ºC and 80 ºC upon addition of instant composition, sample B2, sample C and sample D, respectively.
Figure 5B illustrates increase in the protection factor of fish oil at 60ºC, upon addition of instant composition, sample B2, sample C and sample D, respectively.
Figure 5C illustrates increase in the protection factor of fish oil at 70ºC, upon addition of instant composition, sample B2, sample C and sample D, respectively.
Figure 6A illustrates peroxide value of palm oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 6B illustrates peroxide value of palm oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 6C illustrates para-anisidine value of palm oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 6D illustrates para-anisidine value of palm oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 6E illustrates totox value of palm oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 6F illustrates totox value of palm oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7A illustrates peroxide value of sunflower oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7B illustrates peroxide value of sunflower oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7C illustrates para-anisidine value of sunflower oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7D illustrates para-anisidine value of sunflower oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7E illustrates totox value of sunflower oil at ambient temperature, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 7F illustrates totox value of sunflower oil at 50 ºC, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 8 illustrates peroxide value of fish oil at 10 ºC, upon addition of instant composition and alpha – tocopherol, respectively.
Figure 9 illustrates total polar compound of palm oil, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 10 illustrates total polar compound of sunflolwer oil, upon addition of instant composition, sample A, sample B, sample C, sample D, sample E and sample F, respectively.
Figure 11 illustrates relationship between particle size and antioxidant activity

DETAILED DESCRIPTION OF THE INVENTION
The present disclosure relates to a composition comprising green tea extract and rosemary extract, optionally along with adjuvant or excipient or a combination thereof.

In an embodiment the present disclosure relates to said green tea extract is at a concentration ranging from about 24% w/w to about 80% w/w.
In another embodiment the present disclosure relates to said rosemary extract is at a concentration ranging from about 45%w/w to about 85% w/w.

In yet another embodiment the present disclosure relates to said rosemary extract comprises rosemarinic acid at a concentration ranging from about 0.5%w/w to about 10%w/w, preferably at a range of about 0.5% w/w to about 3.5%w/w and carnosic acid at a concentration ranging from about 1.0%w/w to about 20%w/w, preferably at a range of about 1.5%w/w to about 12%w/w.

In still another embodiment the present disclosure relates to the adjuvant is selected from a group comprising vitamin C, gallic acid, vitamin E, rosmanol, ferulic acid, citric acid, mixed-tocopherol, lecithin, carotenoids and uric acid or any combination thereof.

In still another embodiment the present disclosure relates to the excipient is selected from a group comprising granulating agent, binding agent, lubricating agent, disintegrating agent, sweetening agent, glidant, anti-adherent, anti-static agent, surfactant, coating agent, colouring agent, flavouring agent, plasticizer, suspending agent, additive, emulsifying agent and spheronization agent or any combination thereof.
In still another embodiment the present disclosure relates to the emulsifying agent is polyglyceride fatty acid ester, preferably macrogoglycerol hydroxysterate; and wherein the additive is selected from a group comprising mono-di-glycerides, sorbitol, guar gum and xanthan gum or any combination thereof

In still another embodiment the present disclosure relates to said adjuvant at a concentration ranging from about 1.0w/w to about 10%w/w, preferably ranging from about 1.5%w/w to about 8%w/w.

In still another embodiment the present disclosure relates to said excipient at a concentration ranging from about 0.5% to about 5%w/w, preferably ranging from about 1.0%w/w to about 2.5% w/w.

In still another embodiment the present disclosure relates to particle size of the composition ranging from about 5µm to about 10µm.

In still another embodiment the present disclosure relates to the composition optionally comprising polyphenols at a concentration ranging from about 30% to about 38%.

In still another embodiment the present disclosure relates to oil soluble composition having solubility ranging from about 95% to about 100% with settling less than about 0 about 5%.

The present disclosure further relates to a process of preparing the composition comprising green tea extract and rosemary extract, said process comprising acts of:
a. granulating crude extract of green tea and mixing the granulated extract with the rosemary extract to obtain a mixture;
b. optionally adding adjuvant or excipient or a combination thereof to the mixture; and
c. passing the mixture of step a) or b) through homogenizer to obtain said composition comprising green tea extract and rosemary extract

In an embodiment of the present disclosure, the crude extract of green tea having a particle size of about 50µ to about 200µ is granulated to obtain a extract of particle size ranging from about 10µ to about 20 µ.

In another embodiment of the present disclosure, the composition obtained in step c) has a particle size ranging from about 5µ to about 10µ.

In yet another embodiment of the present disclosure, the crude extract of the green tea are granulated by techniques selected from a group comprising hammer mill, ball mill, plate mill, disc mill, colloid mill, micronisation method, high pressure homogenization and cryogenic grinding or a combination thereof, preferably cryogenic grinding.

In still another embodiment of the present disclosure, the mixing is carried out by an agitator at a speed of about 500rpm to about 1000rpm.

In still another embodiment of the present disclosure, the mixing is at a temperature ranging from about 40 °C to about 65 °C, preferably at about 50 °C for time period ranging from about 2hrs to about 24hrs.

In still another embodiment of the present disclosure, the mixture is homogenized at a pressure ranging from about 500 bar to about 1000 bar.

The present disclosure further relates to a composition comprising the composition as mentioned above and tertiary butyl hydroquinone.
In an embodiment of the present disclosure, the composition as mentioned above is at a concentration ranging from about 50ppm to about 100ppm.

In another embodiment of the present disclosure, the tertiary butyl hydroquinone is at a concentration ranging from about 50ppm to about 100ppm.

The present disclosure further relates to an oil, fat or cosmetic preparation, comprising the composition of claim 1 or claim 20.

The present disclosure further relates to method of preparing the oil, fat or cosmetic preparation as claimed in claim 23, wherein said method comprises act of mixing the composition of claim 1 or claim 17 with the oil, fat or cosmetic ingredients.

The present invention provides an oil soluble composition comprising extract of green tea with rosemary extract, optionally along with adjuvants and excipients, wherein the composition is a natural antioxidant.

In an embodiment, the composition of the instant invention acts as a preservative in edible oil or cooking oil or food grade oil and oils which have cosmetic applications.

In an embodiment, the composition of the instant invention increases the oxidative induction time of the oil, thereby decreasing the rate of primary and secondary oxidation of the oil.

In an embodiment, the composition of the instant invention is added to oil at a concentration as low as about 50ppm to as high as about 1000ppm, unlike the conventionally known synthetic antioxidant, which have an upper limit of 200ppm (under the regulatory guidelines) beyond which it is considered to be carcinogenic, whereas some antioxidant at higher dosage (above 500ppm) acts as a pro-oxidant.

In an embodiment, the composition of the instant invention reduces the formation of polar compounds in the oil, thereby reducing degradation of oil. The composition of the instant invention also enhances the number of frying cycles at elevated temperatures and increases the shelf life of the oil.

In an embodiment, the oils for which the composition of instant invention acts as a preservative are avocado oil, mustard oil, palm oil, peanut oil, rice barn oil, safflower oil, sesame oil, sunflower oil, almond oil, canola oil, coconut oil, corn oil, cottonseed oil, mustard oil, grape seed oil, olive oil, pumpkin seed oil, tea seed oil, walnut oil, fish oil or any combination thereof.

In another embodiment, adjuvant of the instant composition is selected from a group comprising gallic acid, vitamin C, vitamin E, rosmanol, ferulic acid, citric acid, mixed- tocopherol, carotenoids and uric acid or any combination thereof. The excipient of the instant composition is selected from a group comprising granulating agent, binding agent, lubricating agent, disintegrating agent, sweetening agent, glidant, anti-adherent, anti-static agent, surfactant, coating agent, colouring agent, flavouring agent, plasticizer, suspending agent, emulsifying agent and spheronization agent or any combination thereof.

In an embodiment, the composition of the instant invention comprises about 24%w/w to about 80%w/w of green tea extract in powdered form and about 45%w/w to about 80%w/w of rosemary extract in oil form. The composition optionally comprises about 1%w/w to about 10%w/w, preferably from about 1.5% to about 8% w/w of one or more of adjuvants such as vitamin C, gallic acid, vitamin E, rosmanol, ferulic acid, citric acid, mixed- tocopherol, carotenoids, uric acid; optionally along with about 0.5%w/w to about 5%w/w, preferably from about 1.0%w/w to about 2.5%w/w excipients. The green tea extract in the composition of the instant invention is in the form of a powder made through cryogenic grinding with a particle size of about 10µ to about 200µ; and the rosemary is in the form of lipophilic extract containing rosemarinic acid at a concentration ranging from about 1.0% w/w to about 10% w/w, preferably from about 0.5%w/w to about 3.5%w/w and carnosic acid at a concentration ranging from about 1.0% w/w to about 20% w/w, preferably from about 1.5w/w to about 12%w/w.

In an embodiment, the composition of the present invention comprises about 30% to about 38% of polyphenols., wherein the polyphenols content in the crude green tea extract is 90% and when this extract is cryo-grinded, mixed with rosemary extract optionally along with adjuvants and excipients, followed by high pressure homogenizing, the polyphenol content of PRESOL will be reduced to about 30% to about 38%, thereby enhancing the solubility of PRESOL.

In an embodiment, excipients, preferably, emulsifying agents enhances the solubility of the instant composition in oils, wherein the emulsifying agent is macrogoglycerol hydroxystearate. The instant composition comprising macrogogycerol hydroxysterate has a solubility ranging from about 95% to about 100% with settling less than about 5%.

In an embodiment, the instant composition comprises mono-di-glycerides as an additive.
In an embodiment, the composition of present invention is lipid soluble which prevents oxidative rancidity of oils and fats.

In an embodiment, the rosemary extract of the instant composition comprises rosemarinic acid or carnosic acid or a combination thereof.

The present invention further relates to a process of preparing a composition comprising extract of green tea with rosemary extract, optionally along with adjuvants and excipients, wherein said process comprises the following steps:
a) crude green tea extract with a particle size of 50µ to about 200µ is granulated to obtain a extract of particle size ranging from about 10µ to about 20µ;
b) an extract, sage or thyme of a Labiatae herb, preferably rosemary extract, is mixed with granulated green tea extract of particle size 10µ to about 20µ at a temperature ranging from about 40 °C to 65 °C, preferably at about 50 °C for about 2hrs to about 24hrs, in a cylindrical vessel with an agitator rod with paddle that rotates at a speed of about 500 RPM to about 1200 RPM, to form a homogenous solution;
c) optionally adjuvants or excipients or a combination thereof is added to the solution, and mixed thoroughly to obtain a mixture;
d) the mixture of step d) is passed through high pressure homogenizer thrice at different pressures preferably in the range of about 500 bar to about 1000 bar, to obtain the final composition of the present invention.

In an embodiment, the green tea extract is granulated by any one or combination of the method selected from a group comprising hammer mill, ball mill, plate mill, disc mill, colloid mill, micronisation method, high pressure homogenization and cryogenic grinding.

In an embodiment, the crude green tea extract is granulated by cryogenic grinding to a particle size ranging from about 10µ to about 20µ

In an embodiment, the crude green tea extract before grinding is in the form of granular powder with a particle size ranging from about 50µ to about 200µ.

In an embodiment, the adjuvant is selected from a group comprising vitamin C, gallic acid, vitamin E, rosmanol, ferulic acid, citric acid, mixed - tocopherol, carotenoids and uric acid or any combination thereof.

In an embodiment, the excipient is selected from a group comprising granulating agent, binding agent, lubricating agent, disintegrating agent, sweetening agent, glidant, anti-adherent, anti-static agent, surfactant, coating agent, coloring agent, flavoring agent, plasticizer, suspending agent, emulsifying agent and spheronization agent or any combination thereof.

The present invention further relates to fortifying the composition comprising green tea extract and rosemary optionally along with adjuvant or excipients or a combination thereof with antioxidant selected from a group comprising butylated hydroxyanisole, butylated hydroxyl toluene, tertiary butyl hydroquinone, propyl gallate or any combination thereof, preferably butylated hydroxyl toluene.

In an embodiment, fortification of composition comprising green tea extract and rosemary extract optionally along with adjuvant or excipients or a combination thereof with antioxidant selected from a group comprising butylated hydroxyanisole, butylated hydroxyl toluene, tertiary butyl hydroquinone, propyl gallate or any combination thereof is carried out a concentration ranging from about 50ppm to about 200ppm, preferably in the range of about 50ppm to about 100ppm.

In an embodiment, present invention relates to an oil comprising green tea extract and rosemary extract, optionally along with adjuvant or excipients or a combination thereof, wherein the oil is edible oil or cooking oil or food grade oil, oil for cosmetic application, vegetable oil, plant oil or any combination thereof.

In another embodiment, present invention relates to an oil comprising green tea extract and rosemary extract, optionally along with adjuvant or excipients or a combination thereof, fortified with antioxidant selected from a group comprising comprising butylated hydroxyanisole, butylated hydroxyl toluene, tertiary butyl hydroquinone, propyl gallate or any combination thereof, wherein the oil is edible oil or cooking oil or food grade oil, oil for cosmetic application, vegetable oil, plant oil or any combination thereof.

The further embodiment herein describes the scientific and technical terms used in connection with the instant invention and shall have meaning/definitions/equations/glossary that are commonly understood by those skilled in the art:

As used herein, ‘Oil Stability Index (OSI)’ is an American Oil Chemists Society (AOCS) approved method that determines the relative resistance of fat and oil samples to oxidation, which is defined by the following equation-

Oil stability index (OSI) = Induction time of treated oil
Induction time of control oil

As used herein, ‘increase in protection factor’ is defined by the following equation-

Increase in protection factor = OSI of treated oil – OSI of control oil X 100
OSI of control oil

As used herein, ‘PRESOL’ is the final composition of the instant invention, comprising extract of green tea with rosemary extract, optionally along with adjuvants and excipients.

As used herein, ‘control’ is an oil without any antioxidant.

As used herein, ‘Sample A’ is oil with TBHQ (98% activity)

As used herein, ‘Sample B’ is oil with BHA (98% activity)

As used herein, ‘Sample C’ is oil with pulverized green tea (95 % polyphenols)

As used herein, ‘Sample D’ is oil with rosemary extract (8 % Carnosic acid)

As used herein, ‘Sample E’ is oil with cryogrinded green tea (95 % Polyphenols)

As used herein, ‘Sample B1’ is oil with BHA (98% activity)

As used herein, ‘Sample B2’ is oil with alpha tocopherol (96 % activity)

The present invention is further illustrated by the following examples. However, the following examples are provided for illustrative purposes only and are not intended to limit the scope of the invention.

EXAMPLES:

Example 1: Process of preparing PRESOL
a. In order to prepare 1000g of PRESOL, 500g of crude green tea extract with a particle size of about 50µ to about 200µ is cryo-grinded for two cryo cycles for about 10mins each with an intermediate cooling for about 1minute. 470g of green tea extract is recovered from the cryo-grinder, with a particle size of about 10µ to about 20 µ. To 300g of the cryo-grinded green tea extract, about 500g of rosemary extract comprising 11.67% carnosic acid is added and mixed slowly in a cylindrical vessel with an agitator rod with paddle that rotates at a speed of about 500RPM to about 1200RPM at a temperature of about 40ºC to about 65 ºC for about 2hrs to about 24hrs to obtain a mixture. To the mixture, 50g of vitamin C, 50g of Tocopherol, 50g of lecithin, 90g of mono-di-glycerides and 10g of Macrogoglycerol hydroxystearate is added. This obtained mixture is mixed thoroughly and passed through a high pressure homogenizer at three different pressures in the range of about 500bar to about 1000bar to obtain PRESOL with a yield of about 93.5%.

b. In order to prepare 500g of PRESOL, 250g of crude green tea extract with a particle size of about 50µ to about 200 µ is cryo-grinded for two cryo cycles for about 10mins each with an intermediate cooling for about 1minute. 235g of green tea extract is recovered from the cryo-grinder, with a particle size of about 10µ to about 20 µ. To 150g of the cryo-grinded green tea extract, about 250g of rosemary extract comprising 11.67% carnosic acid is added and mixed slowly in a cylindrical vessel with an agitator rod with paddle that rotates at a speed of about 500RPM to about 1200RPM at a temperature of about 40ºC to about 65 ºC for about 2hrs to about 24hrs to obtain a mixture. This mixture is mixed thoroughly and passed through a high pressure homogenizer at three different pressures in the range of about 500bar to about 1000bar to obtain PRESOL with a yield of about 93.5%.

Example 2: Oil Stability index
The oil stability index is determined using the Metrohm’s Rancimat (Metrohm 743 Rancimat).

A. Oil stability index of Palm oil
4.0g of palm oil is weighed into the reaction vessel. The reaction vessel filled with oil is placed in a heating block. The temperature of the block is maintained at 100ºC, 120ºC and 140ºC, respectively. The oil is heated at the said temperatures, followed by passing air at a flow rate of 20L/h. Flowing of air through the oil produces peroxides during primary oxidation and organic acids having low molecular weight, aldehydes and ketones with typical rancid odor during the secondary oxidation phase. These compounds are conveyed by the air flow into measurement vessel containing distilled water where conductivity is being checked continuously by sensor. Variation in conductivity of water shows the presence of organic acids, thus analyzing the induction time of the oil.

The above experiment is repeated by adding 200ppm of TBHQ (sample-A), 200ppm of BHA (sample-B), 200ppm of pulverized green tea extract with 90% polyphenol (sample-C), 200ppm of rosemary (sample-D), 200ppm of green tea extract after cryogenic grinding (sample-E), 200ppm of crude green tea extract (sample-F) and PRESOL at 200ppm, 500ppm and 1000ppm, respectively to palm oil.
From the above experiment, oxidative stability index of untreated oil (control) and treated oil is calculated and the below table illustrates the induction time and the obtained oxidative stability.

Treatment Induction Time (h) Oxidative Stability Index (OSI)
100 °C 120 °C 140 °C 100 °C 120 °C 140 °C
Control PO 41.7 9.64 2.45 1.00 1.00 1.00
Sample A 95.76 20.93 5.26 2.30 2.17 2.15
Sample B 43.33 9.88 2.51 1.04 1.02 1.02
Sample C 74.22 17.95 4.48 1.78 1.86 1.83
Sample D 54.85 13.32 3.14 1.32 1.38 1.28
Sample E 82.46 20.02 4.95 1.98 2.08 2.02
Sample F 58.23 14.17 3.45 1.40 1.47 1.41
PRESOL 63.59 15.57 3.86 1.52 1.62 1.58
PRESOL 97.4 23.74 5.62 2.34 2.46 2.29
PRESOL 135.8 33.64 8.14 3.26 3.49 3.32

Table 1: Oxidative Stability of Palm oil

Based on the obtained oxidative stability index (illustrated in figure 1A), protection factor for the samples are assessed at temperatures 100ºC, 120 ºC and 140 ºC to analyze the increase in the protection factor, which in turn illustrates the enhancement in oxidative stability obtained by treating the oil with PRESOL, TBHQ, BHA, pulverized green tea extract, rosemary extract, green tea extract after cryogenic grinding and crude green tea extract. Figures 1B, 1C and 1D illustrate the increase in the protection factor by treating the oil with PRESOL. Further table 1 illustrates substantial enhancement of oxidative stability of palm oil with PRESOL.

B. Oil stability index of Sunflower oil
4.0g of sunflower oil is weighed into the reaction vessel. The reaction vessel filled with oil is placed in a heating block. The temperature of the block is maintained at 80ºC, 100ºC and 120ºC, respectively. The oil is heated at the said temperatures, followed by passing air at a flow rate of 20L/h. Flowing of air through the oil produces peroxides during primary oxidation and organic acids having low molecular weight, aldehydes and ketones with typical rancid odor during the secondary oxidation phase. These compounds are conveyed by the air flow into measurement vessel containing distilled water where conductivity is being checked continuously by sensor. Variation in conductivity of water shows the presence of organic acids, thus analyzing the induction time of the oil.

The above experiment is repeated by adding 200ppm of TBHQ (sample-A), 200ppm of BHA (sample-B), 200ppm of pulverized green tea extract with 90% polyphenol (sample-C), 200ppm of rosemary extract (sample-D), 200ppm of green tea extract after cryogenic grinding (sample-E), 200ppm of crude green tea extract (sample-F) and PRESOL at 200ppm, 500ppm and 1000ppm, respectively to sunflower oil. From the above experiment, oxidative stability index of untreated oil (control) and treated oil is calculated and the below table illustrates the induction time and the obtained oxidative stability.
Treatment Induction Time (h) Oxidative Stability Index (OSI)
80 °C 100 °C 120 °C 80 °C 100 °C 120 °C
Control SFO 36.48 9.32 2.35 1.00 1.00 1.00
Sample A 109.76 26.82 6.13 3.01 2.88 2.61
Sample B 50.39 13.33 2.64 1.38 1.43 1.12
Sample C 98.52 24.03 5.48 2.70 2.58 2.33
Sample D 52.19 12.94 2.85 1.43 1.39 1.21
Sample E 102.85 25.28 6.05 2.82 2.71 2.57
Sample F 78.42 19.08 4.55 2.15 2.05 1.94
PRESOL 76.52 18.38 4.29 2.10 1.97 1.83
PRESOL 93.18 22.76 5.47 2.55 2.44 2.33
PRESOL 139.72 34.41 8.38 3.83 3.69 3.57

Table 2: Oxidative Stability index of sunflower oil

Based on the obtained oxidative stability index (illustrated in figure 2A), protection factor for the samples are assessed at temperatures 80ºC, 100 ºC and 120 ºC to analyze the increase in the protection factor, which in turn illustrates the enhancement in oxidative stability obtained by treating the oil with PRESOL, TBHQ, BHA, pulverized green tea extract, rosemary extract, green tea extract after cryogenic grinding and crude green tea extract. Figures 2B, 2C and 2D illustrate the increase in the protection factor by treating sunflower oil with PRESOL. Further table 2 illustrates substantial enhancement of oxidative stability of palm oil with PRESOL.

Lower temperature is chosen to study the oxidative stability of sunflower oil, because sunflower is less stable at higher temperatures when compared to palm oil.

C. Oil stability index of Almond Oil
4.0g of almond oil is weighed into the reaction vessel. The reaction vessel filled with oil is placed in a heating block. The temperature of the block is maintained at 100ºC, 120ºC and 140ºC, respectively. The oil is heated at the said temperatures, followed by passing air at a flow rate of 20L/h. Flowing of air through the oil produces peroxides during primary oxidation and organic acids having low molecular weight, aldehydes and ketones with typical rancid odor during the secondary oxidation phase. These compounds are conveyed by the air flow into measurement vessel containing distilled water where conductivity is being checked continuously by sensor. Variation in conductivity of water shows the presence of organic acids, thus analyzing the induction time of the oil.

The above experiment is repeated by adding 200ppm of BHA (sample-B), 200ppm of BHT (sample-B1), 500ppm of alpha tocopherol (sample-B2) and PRESOL at 200ppm, 500ppm and 1000ppm, respectively to almond oil. From the above experiment, oxidative stability index of untreated oil (control) and treated oil is calculated and the below table illustrates the induction time and the obtained oxidative stability.
Treatment Induction Time Oxidative Stability Index (OSI)
100 °C 120 °C 140 °C 100 °C 120 °C 140 °C
Control Almond Oil 22.09 5.06 1.37 1.00 1.00 1.00
Sample B 25.14 6.11 1.52 1.14 1.21 1.11
Sample B1 24.98 5.89 1.48 1.13 1.16 1.08
Sample B2 24.51 5.73 1.43 1.11 1.13 1.04
PRESOL 30.16 7.48 1.77 1.37 1.48 1.29
PRESOL 45.37 11.00 2.65 2.05 2.17 1.93
PRESOL 62.13 15.05 3.62 2.81 2.97 2.64

Table 3: Oxidative Stability index of almond oil

Based on the obtained oxidative stability index (illustrated in figure 3A), protection factor for the samples are assessed at temperatures 100ºC, 120 ºC and 140 ºC to analyze the increase in the protection factor, which in turn illustrates the enhancement in oxidative stability obtained by treating the oil with PRESOL, BHA, BHT and alpha tocopherol. Figures 3B, 3C and 3D illustrate the increase in the protection factor by treating almond oil with PRESOL. Further table 3 illustrates substantial enhancement of oxidative stability of almond oil with PRESOL.

D. Oil stability index of Apricot Oil
4.0g of apricot oil is weighed into the reaction vessel. The reaction vessel filled with oil is placed in a heating block. The temperature of the block is maintained at 100ºC, 120ºC and 140ºC, respectively. The oil is heated at the said temperatures, followed by passing air at a flow rate of 20L/h. Flowing of air through the oil produces peroxides during primary oxidation and organic acids having low molecular weight, aldehydes and ketones with typical rancid odor during the secondary oxidation phase. These compounds are conveyed by the air flow into measurement vessel containing distilled water where conductivity is being checked continuously by sensor. Variation in conductivity of water shows the presence of organic acids, thus analyzing the induction time of the oil.

The above experiment is repeated by adding 200ppm of BHA (sample-B), 200ppm of BHT (sample-B1), 500ppm of alpha tocopherol (sample-B2) and PRESOL at 200ppm, 500ppm and 1000ppm, respectively to apricot oil. From the above experiment, oxidative stability index of untreated oil (control) and treated oil is calculated and the below table illustrates the induction time and the obtained oxidative stability.
Treatment Induction Time at 200 ppm Oxidative Stability Index (OSI)
100 °C 120 °C 140 °C 100 °C 120 °C 140 °C
Control Apricot Oil 26.88 5.87 1.48 1.00 1.00 1.00
Sample B 29.16 6.66 1.57 1.08 1.13 1.06
Sample B1 26.96 5.93 1.49 1.00 1.01 1.01
Sample B2 22.06 5.06 1.32 0.82 0.86 0.89
PRESOL 33.68 8.43 1.88 1.25 1.44 1.27
PRESOL 45.22 10.15 2.46 1.68 1.73 1.66
PRESOL 56.19 13.88 3.29 2.09 2.36 2.22

Table 4: Oxidative Stability index of apricot oil

Based on the obtained oxidative stability index (illustrated in figure 4A), protection factor for the samples are assessed at temperatures 100ºC, 120 ºC and 140 ºC to analyze the increase in the protection factor, which in turn illustrates the enhancement in oxidative stability obtained by treating the oil with PRESOL, BHA, BHT and alpha tocopherol. Figures 4B, 4C and 4D illustrate the increase in the protection factor by treating apricot oil with PRESOL. Further table 4 illustrates substantial enhancement of oxidative stability of apricot oil with PRESOL.

E. Oil stability index of Fish Oil
4.0g of fish oil is weighed into the reaction vessel. The reaction vessel filled with oil is placed in a heating block. The temperature of the block is maintained at 60ºC, 70ºC and 80ºC, respectively. The oil is heated at the said temperatures, followed by passing air at a flow rate of 20L/h. Flowing of air through the oil produces peroxides during primary oxidation and organic acids with low molecular weight, aldehydes and ketones with typical rancid odor during the secondary oxidation phase. These compounds are conveyed by the air flow into measurement vessel containing distilled water where conductivity is being checked continuously by sensor. Variation in conductivity of water shows the presence of organic acids, thus analyzing the induction time of the oil.

The above experiment is repeated by adding, 200ppm of alpha tocopherol (sample-B2), 200ppm of pulverized green tea extract with 90% polyphenol (sample-C), 200ppm of rosemary extract (sample-D) and PRESOL at 200ppm, 500ppm and 1000ppm, respectively to fish oil. From the above experiment, oxidative stability index of untreated oil (control) and treated oil is calculated and the below table illustrates the induction time and the obtained oxidative stability.

Treatment Induction Time Oxidative Stability Index (OSI)
80°C 70°C 60°C 80 °C 70 °C 60 °C
Control Fish oil 2.46 4.98 9.86 1.00 1.00 1.00
Sample B2 6.78 13.74 27.68 2.76 2.76 2.81
Sample D 1.97 4.05 8.36 0.80 0.81 0.85
Sample C 2.28 4.74 9.75 0.93 0.95 0.99
PRESOL at 200 ppm 3.07 6.32 13.04 1.25 1.27 1.32
PRESOL at 500 ppm 4.78 9.68 19.85 1.94 1.94 2.01
PRESOL at 1000 ppm 8.02 16.14 33.07 3.26 3.24 3.35

Table 5: Oxidative Stability index of Fish oil

Based on the obtained oxidative stability index (illustrated in figure 5A), protection factor for the samples are assessed at temperatures 60ºC, 70 ºC and 80 ºC to analyze the increase in the protection factor, which in turn illustrates the enhancement in oxidative stability obtained by treating the oil with PRESOL, alpha tocopherol, pulverized green tea extract with 90% polyphenol and rosemary extract. Figures 5B, 5C and 5D illustrate the increase in the protection factor by treating fish oil with PRESOL. Further table 5 illustrates substantial enhancement of oxidative stability of fish oil with PRESOL.

EXAMPLE 3:
An extension of the Example 1 is provided by a new function known as temperature extrapolation. This is an aid for estimating the shelf life of oils and fats. The extrapolation makes use of the relationship between the measured induction time and the temperature given by van’t Hoff’s law. Several measurements are made at different temperatures and then extrapolated to the storage temperature. The values so obtained allow estimation of the storage stability of the oil or fat containing PRESOL.
The shelf life is calculated using Rancimat shelf life calculator. The shelf life of oil is studied by carrying out chemical analysis, which include studying peroxide value (represents extent of primary oxidation), Para-anisidine value (represents extent of secondary oxidation) and totox value (reflects total oxidation of oil). These parameters are determined for 1kg of untreated oil and treated oil at every 15 days interval for samples stored at ambient temperature and at every 7 days interval for samples stored at 50 °C. The treated oil being, the oil upon addition of 200ppm of TBHQ (sample-A), 200ppm of BHA (sample-B), 200ppm and 500ppm of pulverized green tea extract with 90% polyphenol (sample-C), 200ppm and 500ppm of rosemary extract (sample-D), 200ppm and 500ppm of green tea extract after cryogenic grinding (sample-E), 200ppm and 500ppm of crude green tea extract (sample-F) and PRESOL at 200ppm and 500ppm. The untreated oil is the control sample without any antioxidant.
The threshold limit for per oxide value value under which oil can be used for safe consumption is 10 meq of O2/Kg oil. Similarly, the threshold limit of para-anasidine value and Totox value is 10 and 30, respectively.
A. Determination of Shelf life of palm oil

Treatment Shelf Life (days) at 30 °C
Control 242.9
Sample A 617.0
Sample B 260.9
Sample C 420.8
Sample D 337.8
Sample E 475.1
Sample F 337.4
PRESOL 356.7
PRESOL 606.0
PRESOL 790.8
Table 6: Estimation of Shelf life of Palm oil

Table 6 illustrates that PRESOL suitably enhances the shelf life of palm oil.
Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465 0.465
15 0.948 0.762 0.868 0.904 0.903 0.92 0.941 0.817 0.842 0.61 0.87 0.903 0.77
30 2.053 1.417 1.691 0.955 1.023 0.94 0.968 0.871 0.907 1.018 0.969 0.974 0.85
45 2.838 1.558 2.684 1.015 1.478 0.973 1.104 0.995 1.037 1.488 0.995 1.098 0.949
60 3.523 1.771 4.034 1.956 1.834 1.912 2.074 1.864 1.946 1.893 2.016 2.101 1.753
75 3.879 1.855 4.748 2.626 2.973 2.62 2.793 2.008 2.425 2.714 2.725 2.86 1.823
90 4.51 1.814 5.034 2.635 3.336 2.621 2.673 2.205 2.622 3.154 2.664 2.787 1.856
105 5.893 1.88 5.316 2.68 3.463 2.671 2.67 2.305 2.646 3.347 2.754 2.68 1.934
120 7.163 1.91 6.307 2.735 3.826 2.715 2.763 2.5 2.766 3.613 2.866 2.957 2.076
135 8.15 1.951 7.887 2.764 4.096 2.75 2.807 2.701 2.702 3.776 2.803 2.811 2.143
150 8.807 2.222 8.617 2.798 4.471 2.788 2.901 2.802 2.812 4.051 2.825 2.866 2.224
165 10.481 2.31 11.307 2.886 4.804 2.879 3.092 2.832 2.917 4.717 2.942 3.094 2.309
180 13.712 2.403 12.75 3.229 4.967 3.149 3.505 3.032 3.127 4.787 3.218 3.457 2.501
Table 7: Peroxide value of palm oil at ambient temperature
Table 7 and Figure 6A illustrates that PRESOL at 500 ppm is showing equivalent efficacy in preventing oxidative rancidity as compared to 200 ppm of sample A in palm oil at ambient temperatures. From tables 6 and 7, it is further evident that 500 ppm PRESOL is having efficacy equivalent to 200 ppm Sample A. On the other hand control and sample B are crossing threshold limit after 165 days.

Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482 0.482
7 6.11 1.309 4.735 1.355 2.52 0.743 2.266 1.272 1.358 2.516 0.752 2.249 1.216
15 10.076 1.836 9.973 2.558 3.711 1.63 3.019 2.053 2.606 2.483 1.585 2.867 1.705
22 13.901 1.887 13.062 3.155 4.26 2.077 4.044 2.657 3.046 4.084 2 3.657 1.906
30 17.929 2.005 16.45 4.166 5.485 2.601 4.677 2.693 4.344 3.508 2.584 4.686 2.123
37 36.394 3.074 35.595 5.016 10.528 2.892 5.606 3.016 5.115 6.806 2.806 5.563 2.966
45 61.424 3.686 60.884 6.11 15.1 3.431 6.693 3.903 5.944 9.745 3.387 6.635 3.503
52 50.607 4.102 51.15 6.562 12.969 4.201 7.062 4.485 6.589 9.892 4.027 6.868 4.106
60 30.568 4.491 42.269 7.485 15.711 7.583 8.848 4.891 7.053 11.688 7.482 8.656 4.505
67 24.515 4.75 34.974 8.064 18.284 7.884 10.141 5.159 8.238 13.004 7.49 9.022 4.639
75 15.389 4.893 25.576 9.503 22.92 8.277 11.689 5.714 8.726 14.769 7.945 10.885 4.911
82 10.65 5.209 11.31 14.953 25.251 11.106 17.905 5.955 14.342 24.801 10.292 17.121 5.484
90 12.991 5.395 15.231 19.053 27.907 13.438 22.441 6.502 18.637 26.983 13.188 21.396 5.813
Table 8: Peroxide value of palm oil at 50ºC

Table 8 and figure 6B illustrates that after 90 days of storage at 50ºC, PRESOL at 200 ppm and 500 ppm does not cross threshold limit and are acting similar to sample A.
Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566 2.566
15 3.656 3.062 2.602 3.472 3.458 3.221 3.563 3.032 3.507 3.453 3.25 3.664 2.607
30 3.435 3.045 3.18 3.265 2.569 2.966 3.403 3.077 3.343 3.497 3.367 3.681 2.772
45 2.957 3.113 2.899 3.178 2.603 2.302 3.309 3.096 3.465 3.259 2.562 3.111 2.851
60 2.973 2.615 2.759 3.008 2.74 2.111 3.434 3.003 3.302 3.256 2.459 3.89 2.87
75 3.017 2.731 2.952 3.053 2.86 2.613 3.525 2.923 2.809 3.218 2.662 3.439 3.033
90 3.383 2.772 3.113 3.157 2.946 3.038 3.561 2.831 3.089 2.958 3.024 3.621 3.027
105 3.752 3.217 3.455 3.389 3.347 3.315 3.683 3.022 3.324 3.157 3.184 3.701 3.158
120 3.906 3.303 3.499 3.548 3.502 3.427 3.72 3.136 3.472 3.526 3.338 3.873 3.073
135 4.046 3.438 3.593 3.654 3.626 3.614 3.628 3.256 3.484 3.617 3.493 4.006 3.136
150 4.389 3.473 3.759 3.869 4.049 3.711 4.15 3.33 3.799 3.896 3.697 4.106 3.236
165 5.05 3.639 4.169 3.934 4.231 3.781 4.228 3.468 3.87 4.014 3.835 4.213 3.367
180 5.144 3.808 4.493 4.109 4.363 3.891 4.394 3.764 4.08 4.127 3.944 4.364 3.665
Table 9: Para-anisidine value of palm oil at ambient temperature
Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69 2.69
7 3.143 3.062 3.186 3.006 3.163 3.041 3.169 2.975 2.932 3.048 3.092 3.104 3.117
15 3.585 3.296 4.011 3.337 3.819 3.293 3.417 3.124 3.09 3.329 3.814 3.301 3.437
22 3.686 3.395 4.204 3.449 4.042 3.343 3.595 3.489 3.397 3.462 3.641 3.369 3.608
30 4.545 3.74 5.542 4.033 4.62 4.036 4.121 3.95 3.782 4.066 4.546 4.089 4.273
37 5.062 4.052 5.617 4.303 4.886 4.104 4.694 4.266 4.144 4.337 4.911 4.208 4.727
45 6.054 4.209 5.794 4.636 5.502 4.433 4.771 4.455 4.249 4.606 5.271 4.589 5.085
52 6.41 4.685 5.827 4.835 5.475 4.788 5.075 4.806 4.716 4.807 5.397 4.809 5.121
60 6.942 5.091 6.165 5.079 5.681 4.96 5.205 4.92 4.822 5.137 5.614 5.052 5.155
67 7.035 5.208 6.22 5.423 5.798 5.385 5.674 5.12 4.975 5.501 5.775 5.365 5.639
75 7.29 5.514 6.269 5.271 5.789 5.233 5.417 5.271 5.089 5.3 5.862 5.35 5.454
82 7.36 5.601 6.503 5.942 5.98 5.773 5.996 5.411 5.225 5.877 5.949 5.88 6.118
90 7.676 5.827 6.657 5.956 6.092 5.866 6.048 5.818 5.521 6.047 6.074 5.993 6.096
Table 10: Para-anisidine value of palm oil at 50ºC
Tables 9 and 10 illustrates the para-anisidine value of palm oil with PRESOL, TBHQ, BHA, pulverized green tea extract, crude green tea extract, cryogrinded green tea extract, rosemary extract. From the tables it can be observed that the para-anisidine values are within the threshold limits (10 meq of O2/Kg oil). This is because, para- anisidine values reflects secondary oxidation of oil and since primary oxidation would still be under process, hence secondary oxidation products would not be formed and thereby the para-anisidine values are within the limit.

Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497 3.497
15 5.551 4.585 4.338 5.280 5.265 5.060 5.445 4.666 5.190 4.673 4.991 5.470 4.146
30 7.540 5.879 6.561 5.175 4.615 4.845 5.338 4.818 5.157 5.534 5.305 5.628 4.471
45 8.634 6.228 8.268 5.208 5.560 4.248 5.517 5.086 5.538 6.234 4.552 5.307 4.748
60 10.019 6.157 10.827 6.919 6.408 5.934 7.582 6.731 7.194 7.042 6.490 8.092 6.376
75 10.775 6.441 12.449 8.306 8.807 7.854 9.110 6.939 7.659 8.647 8.112 9.159 6.678
90 12.403 6.400 13.181 8.427 9.617 8.279 8.907 7.241 8.332 9.267 8.352 9.195 6.738
105 15.539 6.976 14.088 8.749 10.273 8.656 9.023 7.631 8.615 9.851 8.692 9.062 7.026
120 18.232 7.122 16.114 9.018 11.154 8.857 9.245 8.137 9.004 10.753 9.070 9.787 7.225
135 20.347 7.339 19.368 9.183 11.817 9.113 9.241 8.658 8.889 11.169 9.099 9.629 7.422
150 22.002 7.916 20.994 9.466 12.991 9.286 9.953 8.934 9.424 11.998 9.347 9.839 7.683
165 26.012 8.258 26.783 9.707 13.839 9.539 10.411 9.133 9.705 13.448 9.718 10.402 7.984
180 32.567 8.614 29.993 10.567 14.297 10.188 11.404 9.828 10.335 13.701 10.380 11.278 8.666
Table 11: Totox value of palm oil at ambient temperature
Table 11 and figure 6E illustrates that control and sample B has crossed the threshold limit after 180 days as in case of peroxide value. On the other hand PRESOL at 500 ppm and Sample A at 200 ppm are performing equally.
Day Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655 3.655
15 15.364 5.679 12.656 5.716 8.203 4.528 7.701 5.520 5.764 8.124 4.608 7.615 5.364
30 23.737 6.967 23.957 8.454 11.242 6.553 9.454 7.230 8.542 8.780 6.472 9.170 6.500
45 31.488 7.169 30.328 9.759 12.562 7.498 11.683 8.803 9.553 11.809 7.369 10.922 7.210
60 40.404 7.750 38.442 12.365 15.590 9.239 13.475 9.336 12.755 11.563 9.257 13.644 8.028
75 77.850 10.200 76.806 14.336 25.942 9.889 15.906 10.298 14.567 18.523 9.820 15.852 10.077
90 128.903 11.581 127.563 16.856 35.701 11.295 18.157 12.261 16.494 24.761 11.362 18.354 11.256
105 107.624 12.888 108.126 17.959 31.412 13.190 19.199 13.775 17.984 25.182 12.863 18.857 12.929
120 68.077 14.074 90.702 20.048 37.103 20.125 22.902 14.702 19.243 28.990 20.016 22.468 13.833
135 56.064 14.709 76.168 21.551 42.365 21.152 25.957 15.439 21.976 31.783 20.345 23.683 14.255
150 38.068 15.300 57.420 24.276 51.629 21.786 28.796 16.699 22.753 35.399 21.239 27.223 14.912
165 28.660 16.019 29.123 35.849 56.482 27.985 41.807 17.322 34.561 55.551 26.465 40.359 16.193
180 33.658 16.617 37.118 44.062 61.906 32.742 50.931 18.821 43.321 60.041 32.369 48.888 17.146
Table 12: Totox value of palm oil at 50ºC
Table 12 and figure 6F illustrates that control and sample B has crossed the threshold limit after 180 days as in case of peroxide value. On the other hand PRESOL at 500 ppm and Sample A at 200 ppm are performing equally.

B. Determination of Shelf life of sunflower oil
Treatment Shelf Life (days) at 30 °C
Control SFO 46.3
Sample A at 200 ppm 171.0
Sample B at 200 ppm 87.2
Sample C at 200 ppm 137.6
Sample Dat 200ppm 83.2
Sample Eat 200ppm 147.8
Sample Fat 200ppm 115.7
PRESOL at 200 ppm 117.5
PRESOL at 500 ppm 134.2
PRESOL at 1000 ppm 198.5
Table 13: Estimation of Shelf life of sunflower oil

Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758 0.758
15 4.368 2.008 3.636 3.551 4.285 3.446 3.618 2.687 4.232 4.164 3.945 4.38 2.426
30 13.706 2.927 8.816 5.794 8.638 5.292 6.656 3.478 6.862 8.44 6.027 6.985 3.096
45 19.876 3.223 10.813 8.277 9.698 7.17 8.878 4.076 10.058 9.205 8.654 10.858 3.541
60 27.018 3.335 14.091 10.01 13.774 9.427 12.059 4.687 12.682 13.552 11.797 14.07 4.007
75 32.633 3.711 17.632 11.944 16.788 11.138 13.25 5.269 13.733 15.923 12.073 14.205 4.353
90 39.991 4.088 22.615 13.127 21.797 12.531 14.842 6.462 16.117 20.813 13.58 17.56 4.8
105 48.713 4.77 28.884 17.58 26.33 15.304 19.278 7.312 19.456 25.293 16.844 20.159 5.486
120 57.501 5.13 33.526 22.001 28.554 20.025 24.212 8.328 23.059 27.932 20.989 25.356 6.176
135 67.881 5.809 41.168 25.038 35.363 23.046 28.144 9.478 25.936 34.289 23.77 29.161 6.838
150 77.186 6.387 45.546 30.001 40.412 27.58 33.795 10.037 31.597 39.268 28.875 35.537 7.648
165 86.946 7.875 55.612 35.109 51.02 33.144 37.494 11.674 36.1 49.919 34.131 39.538 8.613
180 92.913 8.252 60.578 39.591 54.15 37.852 42.26 12.461 40.22 52.295 38.821 44.094 9.462
Table 14: Peroxide value of sunflower oil at ambient temperatures
Table 14 and figure 7A illustrates that that PRESOL at 200 ppm and PRESOL at 500 ppm have better activity than Samples B, C, D, E and F. Control sample is observed to cross threshold limit within one month and other samples apart from PRESOL and sample A have crossed the threshold limit in two months. Further, PRESOL at 500ppm is showing similar activity for preventive oxidative rancidity as compared to 200 ppm of TBHQ.
Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807 0.807
7 6.862 1.293 6.172 3.01 4.663 2.964 3.615 2.01 3.388 4.458 3.161 3.805 1.607
15 12.413 2.49 11.596 4.483 6.079 4.456 5.065 2.892 4.89 5.865 4.645 5.472 2.631
21 16.064 5.051 15.146 7.447 8.043 7.022 8.066 6.387 7.67 7.905 7.2 8.199 5.183
30 23.001 9.989 18.486 9.431 12.125 9.556 11.001 9.053 9.722 12.034 9.656 11.413 8.456
37 29.478 12.472 23.087 15.431 15.066 12.375 17.042 10.425 15.522 14.949 12.584 17.172 9.91
Table 15: Peroxide value of sunflower oil at 50ºC
Table 15 and figure 7B illustrates that all the samples apart from PRESOL is crossing the threshold limit within one month. Since sunflower oil is unstable at higher temperature and is more susceptible to oxidation at higher temperature, PRESOL is showing better activity in preventing oxidative rancidity of sunflower oil up to 37 days.
Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941 5.941
15 6.217 6.035 6.166 6.107 6.186 6.262 6.144 6.075 6.112 6.141 6.264 6.116 6.024
30 6.29 6.083 6.258 6.252 6.212 6.378 6.286 6.142 6.254 6.208 6.364 6.301 6.102
45 6.443 6.141 6.452 6.331 6.311 6.297 6.382 6.17 6.334 6.293 6.306 6.366 6.152
60 7.115 6.224 6.793 6.702 6.37 6.405 6.792 6.256 6.772 6.354 6.453 6.836 6.23
75 7.324 6.326 6.85 6.733 6.505 6.554 6.822 6.376 6.709 6.507 6.578 6.853 6.335
90 7.771 6.493 6.962 7.083 6.73 6.651 7.222 6.556 7.119 6.704 6.693 7.242 6.513
105 8.101 6.712 7.381 7.492 7.169 7.022 7.601 6.773 7.513 7.236 7.037 7.686 6.746
120 8.352 7.039 8.154 7.885 7.583 7.729 8.053 7.109 8.006 7.585 7.853 8.15 7.056
135 9.794 7.482 9.748 8.106 7.859 9.22 8.532 7.567 8.126 7.819 9.234 8.551 7.538
150 10.698 7.901 10.661 9.029 8.07 8.724 9.467 7.903 9.109 8.057 8.756 9.484 7.935
165 11.13 8.408 11.013 10.605 8.633 10.551 10.85 8.534 10.652 8.613 10.692 10.904 8.466
180 13.065 8.896 12.115 10.803 9.136 10.767 11.164 9.046 10.827 9.123 10.772 11.211 8.982
Table 16: Para-anisidine value of sunflower oil at ambient temperature

Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237 6.237
7 6.511 6.334 6.439 6.435 6.483 6.262 6.374 6.384 6.453 6.444 6.285 6.403 6.348
15 7.165 6.675 6.842 6.548 6.51 6.378 6.636 6.706 6.555 6.499 6.401 6.689 6.681
21 7.926 7.026 7.347 6.623 6.608 6.297 6.742 7.068 6.688 6.54 6.338 6.805 7.05
30 8.532 7.11 7.392 6.702 6.37 6.405 6.729 7.091 6.82 6.36 6.467 6.923 7.063
37 9.28 7.503 8.035 6.733 6.505 6.554 6.792 7.383 6.781 6.492 6.62 6.897 7.383
45 10.014 7.754 8.438 7.083 6.73 6.651 7.222 7.558 7.255 6.707 6.756 7.372 7.504
52 10.775 8.016 9.162 7.492 7.169 7.022 7.698 7.697 7.509 7.149 7.104 7.841 7.538
60 11.464 8.208 9.641 7.885 7.583 7.729 8.112 7.718 7.964 7.593 7.761 8.132 7.597
67 12.435 8.506 9.748 8.693 7.859 8.924 8.85 8.097 8.724 7.813 9.014 8.793 7.949
75 13.278 8.863 10.661 9.613 9.249 9.313 9.763 8.182 9.696 9.328 9.386 9.815 8.06
82 14.388 8.945 11.013 10.605 10.109 10.551 11.195 8.526 10.63 10.084 10.692 11.275 8.38
90 15.714 9.484 12.411 11.098 11.188 11.061 11.462 9.471 11.146 11.319 11.059 11.611 9.378
Table 17: Para-anisidine value of sunflower oil at 50ºC
Table 16, table 17 and figure 7C illustrates that PRESOL is having better activity when compared to oils of samples B, C, D, E and F and it is having activity similar to oil comprising TBHQ.
Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458 7.458
15 14.953 10.051 13.438 13.210 14.756 13.154 13.380 11.449 14.577 14.469 14.153 14.877 10.877
30 33.701 11.937 23.889 17.839 23.488 16.962 19.598 13.098 19.977 23.089 18.417 20.270 12.294
45 46.195 12.588 28.077 22.885 25.708 20.637 24.138 14.323 26.450 24.702 23.615 28.082 13.234
60 61.152 12.893 34.976 26.722 33.918 25.258 30.909 15.629 32.137 33.457 30.046 34.977 14.243
75 72.589 13.749 42.114 30.622 40.080 28.831 33.322 16.914 34.175 38.353 30.725 35.262 15.042
90 87.753 14.669 52.191 33.337 50.324 31.714 36.906 19.479 39.354 48.329 33.854 42.363 16.113
105 105.528 16.253 65.149 42.652 59.829 37.629 46.156 21.396 46.424 57.823 40.725 48.004 17.717
120 123.353 17.299 75.207 51.888 64.690 47.780 56.477 23.765 54.124 63.450 49.832 58.862 19.408
135 145.557 19.100 92.085 58.183 78.584 55.311 64.820 26.522 59.997 76.398 56.775 66.872 21.215
150 165.070 20.674 101.75 69.030 88.894 63.884 77.056 27.976 72.303 86.593 66.505 80.558 23.230
165 185.022 24.158 122.23 80.822 110.674 76.838 85.838 31.882 82.851 108.45 78.953 89.981 25.692
180 198.890 25.399 133.27 89.984 117.436 86.471 95.684 33.968 91.268 113.71 88.413 99.400 27.906
Table 18: Totox value of sunflower oil at ambient temperature
Day Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Sample C at 500 ppm Sample D at 500 ppm Sample E at 500 ppm Sample F at 500 ppm PRESOL at 500 ppm
0 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850 7.850
7 20.235 8.919 18.784 12.456 15.810 12.190 13.603 10.403 13.229 15.359 12.607 14.013 9.563
15 31.991 11.655 30.034 15.515 18.669 15.291 16.765 12.490 16.335 18.229 15.692 17.633 11.944
22 40.055 17.128 37.639 21.518 22.695 20.341 22.873 19.842 22.028 22.350 20.738 23.203 17.416
30 54.533 27.087 44.365 25.564 30.620 25.517 28.731 25.197 26.265 30.428 25.780 29.748 23.975
37 68.235 32.447 54.210 37.595 36.637 31.304 40.876 28.234 37.826 36.390 31.789 41.240 27.204
45 84.177 36.287 68.642 46.012 46.960 35.568 47.348 34.882 46.492 46.600 36.207 49.221 34.344

Table 19: Totox value of sunflower oil at 50ºC
Table 18 and figure 7E illustrates that 200ppm and 500ppm of PRESOL is having better activity than antioxidant in samples B, C, D, E and F. Further, control is observed to cross threshold limit within one month. 200ppm PRESOL is observed to prevent oxidative rancidity of sunflower oil up to 165 days and 500ppm PRESOL is observed to prevent oxidative rancidity of sunflower oil up to 180 days. From this illustration, it can inferred that PRESOL concentration can be increased accordingly in order to prevent oxidative rancidity of oils, whereas the concentration of synthetic antioxidants cannot be increased over a particular limit as per the regulatory guidelines as they might cause harmful effects. For instance, TBHQ is a synthetic antioxidant, which cannot be used beyond the concentration of 200ppm, as it is considered to be carcinogenic beyond 200ppm. Further form the table 19 and figure 7F, it can be observed that the totox value of sunflower oil with PRESOL (200ppm and 500ppm) does not cross the threshold limit of 30 till 37 days, whereas sunflower oil with all other antioxidants (samples A, B, C, D, E, F) crosses the threshold limit by 37 days.
C. Determination of Shelf life of Fish oil
Day Control Fish Oil Alpha Toco (Sample A1) at 200 ppm PRESOL at 200 ppm
0 1.74 1.74 1.74
3 2.09 2.13 1.89
6 3.26 3.33 2.73
9 4.62 4.71 3.46
12 5.39 5.41 3.97
15 6.74 6.82 4.39
18 7.69 7.84 4.85
21 8.98 9.02 5.48
24 10.06 10.32 5.74
27 11.38 12.02 6.24
30 12.97 13.01 6.68
33 15.39 15.44 7.14
36 16.98 17.16 7.72
39 18.72 18.94 8.26
42 20.63 20.86 8.63
45 24.82 25.09 9.01
48 28.63 28.71 9.45
51 34.74 33.89 9.87
54 39.42 39.01 10.23
57 44.66 44.14 10.69
60 48.95 49.22 11.57
63 48.05 50.06 12.03
66 46.82 51.22 12.84
69 45.08 50.85 13.38
72 41.86 50.13 14.05
75 39.71 49.72 14.87
Table 20: Peroxide value of fish oil at 10ºC
Table 20 and figure 8 illustrates that peroxide value of control fish oil and fish oil added with alpha tocopherol (200ppm) crosses the threshold limit of 8meq/kg after 21 days, whereas fish oil with 500ppm of PRESOL is does not cross the threshold limit till 39 days of storage, hence, proving PRESOL to be more powerful antioxidant than synthetic antioxidants.
D. Determination of Shelf life of canola oil in the presence of PRESOL
Shelf life of canola oil is analyzed upon adding 200ppm of TBHQ (sample-A), BHA (sample-B), green tea extract with 90% polyphenol (sample-C), rosemary extract (sample-D), green tea extract after cryogenic grinding (sample-E), and PRESOL, respectively to canola oil.
Sample Shelf Life (No. Of Days)
Control (Canola Oil) 58
Sample-A 181
Sample-B 37
Sample-C 149
Sample-D 74
Sample-E 202
PRESOL 256
Table 21

From the table 21, it is illustrated that PRESOL increases the shelf life of canola oil by enhancing oxidative stability of the oil.

Example 4: Frying cycle analysis

Palm oil and sunflower oil with antioxidants added to it are obtained from a local refinery. Frying studies are carried out with control oil and by adding TBHQ (Sample A), BHA (Sample B), GT Pulverized (Sample C), ROS (Sample D), GT Cryo grinded (Sample E), GT Crude Extract (Sample F) and PRESOL, respectively to the oils. Experiments are carried out a concentration of 200ppm and 500 ppm. Fresh potatoes of less reduced sugar content variety are used throughout the experiment which is purchased form a local supermarket. Oil samples are withdrawn from the fryer at the end of every day and stored at -4ºC until it is tested for quality parameters viz, total polar compounds. Polar compounds present in oil and fats are measured by column chromatography using standard method, ES ISO 8420:2012.
A. Frying cycle analysis of Palm oil

Cycle Control PO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Threshold Limit
0 6.057 6.307 6.325 6.357 6.752 6.304 6.655 6.528 27
8 10.275 9.18 9.754 9.087 11.039 8.944 9.703 8.564 27
16 17.083 13.4 15.964 12.322 15.534 12.071 13.17 10.565 27
24 21.268 17.913 19.329 15.872 17.699 15.127 16.264 13.855 27
32 26.9 23.217 25.264 20.351 23.851 19.55 23.583 15.064 27
40 31.33 28.187 30.192 27.059 30.051 26.005 28.072 18.058 27

Table 22: Total polar compounds of Palm oil

Table 22 illustrates that palm oil with TBHQ, BHA, GT Pulverized, ROS, GT Cryo grinded and GT Crude Extract have crossed threshold limit for total polar compounds after 40th cycle except oil with PRESOL.

B. Frying cycle analysis of sunflower oil

Cycle Control SFO sample A at 200 ppm Sample B at 200 ppm Sample C at 200 ppm Sample D at 200ppm Sample E at 200 ppm Sample F at 200 ppm PRESOL at 200 ppm Threshold Limit
0 3.616 3.746 3.515 3.497 3.872 3.763 3.763 3.856 27
8 7.45 6.375 6.958 6.079 6.888 6.244 6.742 6.938 27
16 15.037 13.632 14.124 9.548 13.986 9.048 11.47 9.023 27
24 21.025 18.505 20.105 13.17 19.39 13.062 16.181 11.616 27
32 27.194 23.123 26.085 20.137 25.589 19.98 22.511 15.584 27
40 33.079 28.648 31.692 27.00 30.073 26.182 27.574 19.156 27

Table 23: Total Polar compounds of sunflower oil

Table 23 illustrates that sunflower oil with TBHQ, BHA, GT Pulverized, ROS, GT Cryo grinded and GT Crude Extract have crossed threshold limit for total polar compounds after 40th cycle except oil with PRESOL.

Example 4: Effect Of Particle Size On Antioxidant Activity Of Vegetable Oils

Effect of particle size analysis on antioxidant activity is analysed with the help of Rancimat (Metrohm 743) which gives oxidative stability index based on the induction time (h). Rancimat analysis is carried out at two different temperature in order to set relationship between induction time and temperature of the studies.
Table 24 shows the particle size of different samples and its effect on antioxidant activity. In this, we have analyzed the particle size of samples with the help of Particle Size Analyzer (Microtrac, S3500) and its antioxidant activity with the help of Rancimat (Metrohm 743). Table 24 shows that reducing the particle size of GT extract has an impact on antioxidant activity. It is observed that when the particle size of GT is reduced, the Rancimat induction time increases which proves that the antioxidant activity increases on reduction of particle size. Further, the optimum particle size is between 5 to 10µm at which the maximum activity is achieved.
Treatment Particle Size (µm) Induction Period (h) of PO at
120 °C Induction Period (h) of SFO 100 °C
Control 9.64 9.32
Sample A Crystalline form 20.93 26.82
Sample B Crystalline form 9.88 13.33
Sample C 46.48 17.95 24.03
Sample D Dispersed form 13.32 12.94
Sample E 12.81 20.02 25.28
Sample F 119.3 14.17 19.08
PRESOL 6.12 15.57 18.38
PRESOL 6.12 23.74 22.76
PRESOL 6.12 33.64 34.41
Table 24: Relationship between particle size and antioxidant activity of TBHQ (sample-A), 200ppm of BHA (sample-B), 200ppm of pulverized green tea extract with 90% polyphenol (sample-C), 200ppm of rosemary extract (sample-D), 200ppm of green tea extract after cryogenic grinding (sample-E), 200ppm of crude green tea extract and PRESOL at 200ppm, 500ppm and 1000ppmn in palm oil (PO) and sunflower oil (SFO), respectively.

From table 24 and figure 11 it is illustrated that reducing particle size increases the solubility and aids to prevent sedimentation of the active components in oil matrix. As observed in the table, PRESOL at lower particle size is able to show better antioxidant activity thereby reducing the oxidative rancidity of the oil.

Example 6: Fortification of tertiary butyl hydroquinone (TBHQ) and PRESOL:
Synergism of TBHQ and PRESOL is analyzed in palm oil. If the frying industry is considered, synthetic antioxidant TBHQ is being added to the oils. However, there is a limitation of adding TBHQ i.e. not above 200ppm, due to which frying oils cannot be used for longer frying cycles. In order to overcome this limitation, TBHQ is fortified with PRESOL and added to vegetable oils in order to increase the frying cycles which ultimately increases the shelf life of end products. Fortification of TBHQ & PRESOL is analyzed in Rancimat at different concentrations, ranging from about 50 to 200ppm in palm oil. The below table 25 illustrates all the combination of fortification of TBHQ and PRESOL used in palm oil to illustrate maximum activity of fortified combination.
Synergism is calculated based on the below equation:
Synergism (%) = (IPX1X2 - IPo) – [(IPX1 - IPo) + (IPX2 - IPo)] X 100
[(IPX1 - IPo) + (IPIPX2 - IPo)]

Where,
IP0 = Induction Period for Control Oil
IPX1 = Induction Period for X1 (TBHQ)
IPX2 = Induction Period for X2 (PRESOL)
IPX1x2 = Induction Period for combination of X1 (TBHQ) and X2 (PRESOL)

Vegetable Oil TBHQ (Sample A)(ppm)
X1 PRESOL (Sample G)(ppm)
X2 Induction Time (H) at 120 C increase in induction time as compared to control (X-C) OSI Increase in OSI (%) Synergism (%)
PO Control (C) 0 0 9.08 0.00 1
PO 50 0 9.57 0.49 1.054 5.12
PO 100 0 11.01 1.93 1.21 17.53
PO 200 0 14.24 5.16 1.57 36.24
PO 0 50 10.52 1.44 1.16 13.69
PO 0 100 11.36 2.28 1.25 20.07
PO 0 200 15.57 6.49 1.71 41.68
PO 200 100 31.8 22.72 3.50 71.45 205.38
PO 100 200 25.71 16.63 2.83 64.68 97.51
PO 100 100 30.05 20.97 3.31 69.78 398.10
PO 50 50 11.63 2.55 1.28 21.93 32.12
PO 200 200 29.32 20.24 3.23 69.03 73.73
PO 50 100 20.34 11.26 2.24 55.35 306.50
PO 50 200 24.09 15.01 2.65 62.31 115.04
PO 100 50 18.5 9.42 2.04 50.92 179.53
PO 200 50 22.44 13.36 2.47 59.54 102.42
Table 25: Synergism of TBHQ and PRESOL combination.

Table 25 illustrates that the combination of 100 ppm of TBHQ and 100ppm of PRESOL is showing synergistic antioxidant activity by suitably enhancing the oxidative stability of palm oil.

Example 7: Solubility of PRESOL in sunflower oil.

Solubility of PRESOL is tested with respect to pulverized green tea extract (sample-C), green tea extract cryogrinded (sample-E) and crude green tea (sample-F) in sunflower oil. Solubility test is carried out by analyzing the settling percentage in the oil over storage. Settling is determined by analyzing the polyphenol content, which is present in the oil comprising the above mentioned samples using spectrophotometer at 540nm.

Trial No Treatments Concentration of Blends in ppm Concentration Polyphenols in blends ppm Concentration of Polyphenols in Oil during storage(ppm) %Solubility %Settling
1 PRESOL 200 41.86 41.35 98.78 1.22
2 500 104.65 102.54 97.98 2.02
3 1000 209.3 200.85 95.96 4.04
4 Sample C 200 190 68.59 36.1 63.9
5 500 475 147.96 31.15 68.85
6 1000 950 253.175 26.65 73.35
7 Sample E 200 190 80.199 42.21 57.79
8 500 475 179.55 37.8 62.2
9 1000 950 314.45 33.1 66.9
10 Sample F 200 190 48.64 25.6 74.4
11 500 475 85.56 18.01 81.99
12 1000 950 99.37 10.46 89.54
Table 26: Solubility of PRESOL in sunflower oil.

Total Polyphenols % = Wt. of std ×10 × Abs. of sample × Ds × P × 100
100 50 Abs. of std
Where ,
Ds = dilution of sample
Wt= Weight of the sample in gram
P= Purity of standard

From the table 26, it is illustrated that settling is on the higher side for green tea extract without grinding (sample-F) followed by pulverized green tea extract (sample-C) and at a lower side for green tea extract with cryogrinding (sample-E). However, PRESOL is having highest solubility percentage in the oil with very minimal settling over storage. The high solubility of PRESOl with minimal settling is directly linked to the percentage of polyphenols present in the composition. Though Polyphenols is the active component which provides the antioxidant activity in green tea extracts, it has very poor solubility in oil. On the other hand, PRESOL with polyphenol percentage of about 30% to about 38% shows enhanced antioxidant activity with minimal settling over storage.

Example 8- Determining solubility of PRESOL with different emulsifying agents

Trial No Treatments

Emulsifying Agent Concentration of Blends in ppm Concentration Polyphenols in blends ppm Concentration of Polyphenols in Oil during storage(ppm) %Solubility %Settling
1 PRESOL Macrogolglycerol Hydroxystearate 200 41.86 41.35 98.78 1.22
2 500 104.65 102.54 97.98 2.02
3 1000 209.3 200.85 95.96 4.04
4 PRESOL
Sorbitol ( glucitol) 200 41.86 27.26 65.11 34.89
5 500 104.65 66.02 63.09 36.91
6 1000 209.3 125.33 59.88 40.12
7 PRESOL
DATEM (diacetyl tartaric acid ester of mono- and diglycerides) 200 41.86 33.17 79.25 20.75
8 500 104.65 80.01 76.45 23.55
9 1000 209.3 146.68 70.08 29.92
10 PRESOL
Guar gum (Galactomannan) 200 41.86 25.76 61.53 38.47
11 500 104.65 60.94 58.23 41.77
12 1000 209.3 114.38 54.65 45.35

Table 27- Solubility of PRESOL comprising emulsifying agents
Example 9 – Cost Advantage of PRESOL over TBHQ
Case 9.1 with 200 ppm TBHQ in Sunflower Oil (SFO)
Consider a fryer with oil bath capacity of 100 litres. We are using SFO as our medium of oil for frying potato chips. Rate of oil is 90/- per kg. Rate of TBHQ is approx. 750/- per kg. TBHQ dosage is 200ppm, which is 0.2 gms in 1000gms of oil i.e. 0.15 rupees per kg. So the net rate of oil will be (90 + 1.5) = 90.15/- per kg. Number of batches fried after adding 200ppm TBHQ is 40 batches. Hence, total cost incurred per batch of frying will be, 100*90.15/40 = 225.375/- per batch.
Case 9.2 with 200 ppm PRESOL in Sunflower Oil (SFO)
Consider a fryer with oil bath capacity of 100 litres. We are using SFO as our medium of oil for frying potato chips. Rate of oil is 90/- per kg. Rate of PRESOL is approx. 2500/- per kg. PRESOL dosage is 200ppm, which is 0.2 gms in 1000gms of oil i.e. 0.5 rupees per kg. So the net rate of oil will be (90 +0.5) = 90.5/- per kg. Number of batches fried after adding 200ppm PRESOL is 62 batches. Hence, total cost incurred per batch of frying will be, 100*90.5/62 = 146.97/- per batch.
Cost saved is about rupees78/- (Approx. 35% reduction per Batch).
Case 9.3 with 200 ppm TBHQ in Palm Oil (PO)
Consider a fryer with oil bath capacity of 100 litres. We are using PO as our medium of oil for frying potato chips. Rate of oil is 50/- per kg. Rate of TBHQ is approx. 750/- per kg. TBHQ dosage is 200ppm, which is 0.2 gms in 1000gms of oil i.e. 0.15 rupees per kg. So the net rate of oil will be (50 + 0.15) = 50.15/- per kg. Number of batches fried after adding 200ppm TBHQ is 40 batches. Hence, total cost incurred per batch of frying will be, 100*50.15/40 = 125.375/- per batch.
Case 9.4 with 200 ppm PRESOL in Palm Oil (PO)
Consider a fryer with oil bath capacity of 100 litres. We are using PO as our medium of oil for frying potato chips. Rate of oil is 50/- per kg. Rate of PRESOL is approx. 2500/- per kg. PRESOL dosage is 200ppm, which is 0.2 gms in 1000gms of oil i.e. 0.5 rupees per kg. So the net rate of oil will be (50 + 5) = 50.5/- per kg. Number of batches fried after adding 200ppm PRESOL is 72 batches. Hence, total cost incurred per batch of frying will be, 100*50.5/72 =70.14/- per batch.
Cost saved is about rupees 55/- (Approx. 44% reduction per Batch).
Although cost is a relative parameter which changes with time, the costs showcased herein are applicable in August 2013, and are presented to provide an idea of the cost differential between the composition of the instant invention and the most used sample, TBHQ. This relative differential will remain the same
,CLAIMS:1. A composition comprising green tea extract and rosemary extract, optionally along with adjuvant or excipient or a combination thereof.
2. The composition as claimed in claim 1, wherein said green tea extract is at a concentration ranging from about 24% w/w to about 80% w/w.
3. The composition as claimed in claim 1, wherein said rosemary extract is at a concentration ranging from about 45%w/w to about 85% w/w.
4. The composition as claimed in claim 1, wherein said rosemary extract comprises rosemarinic acid at a concentration ranging from about 0.5%w/w to about 10%w/w, preferably at a range of about 0.5% w/w to about 3.5%w/w and carnosic acid at a concentration ranging from about 1.0%w/w to about 20%w/w, preferably at a range of about 1.5%w/w to about 12%w/w.
5. The composition as claimed in claim 1, wherein the adjuvant is selected from a group comprising vitamin C, gallic acid, vitamin E, rosmanol, ferulic acid, citric acid, mixed-tocopherol, lecithin, carotenoids and uric acid or any combination thereof.
6. The composition as claimed in claim 1, wherein the excipient is selected from a group comprising granulating agent, binding agent, lubricating agent, disintegrating agent, sweetening agent, glidant, anti-adherent, anti-static agent, surfactant, coating agent, colouring agent, flavouring agent, plasticizer, suspending agent, additive, emulsifying agent and spheronization agent or any combination thereof.
7. The composition as claimed in claim 6, wherein the emulsifying agent is polyglyceride fatty acid ester, preferably macrogoglycerol hydroxysterate; and wherein the additive is selected from a group comprising mono-di-glycerides, sorbitol, guar gum and xanthan gum or any combination thereof
8. The composition as claimed in claim 5, wherein said adjuvant is at a concentration ranging from about 1.0w/w to about 10%w/w, preferably ranging from about 1.5%w/w to about 8%w/w.
9. The composition as claimed in claim 6, wherein said excipient is at a concentration ranging from about 0.5% to about 5%w/w, preferably ranging from about 1.0%w/w to about 2.5% w/w.
10. The composition as claimed in claim 1, wherein particle size of the composition ranges from about 5µm to about 10µm.
11. The composition as claimed in claim 1, wherein the composition optionally comprises polyphenols at a concentration ranging from about 30% to about 38%.
12. The composition as claimed in claim1 is oil soluble having solubility ranging from about 95% to about 100% with settling less than about 0 about 5%.
13. A process of preparing the composition comprising green tea extract and rosemary extract, said process comprising acts of:
d. granulating crude extract of green tea and mixing the granulated extract with the rosemary extract to obtain a mixture;
e. optionally adding adjuvant or excipient or a combination thereof to the mixture; and
f. passing the mixture of step a) or b) through homogenizer to obtain said composition comprising green tea extract and rosemary extract
14. The process as claimed in claim 13, wherein the crude extract of green tea having a particle size of about 50µ to about 200µ is granulated to obtain a extract of particle size ranging from about 10µ to about 20 µ.
15. The process as claimed in claim 13, wherein the composition obtained in step c) has a particle size ranging from about 5µ to about 10µ.
16. The process as claimed in claim 13, wherein the crude extract of the green tea is granulated by techniques selected from a group comprising hammer mill, ball mill, plate mill, disc mill, colloid mill, micronisation method, high pressure homogenization and cryogenic grinding or a combination thereof, preferably cryogenic grinding.
17. The process as claimed in claim 13, wherein the mixing is carried out by an agitator at a speed of about 500rpm to about 1000rpm.
18. The process as claimed in claim 17, wherein the mixing is at a temperature ranging from about 40 °C to about 65 °C, preferably at about 50 °C for time period ranging from about 2hrs to about 24hrs.
19. The process as claimed in claim 13, wherein the mixture is homogenized at a pressure ranging from about 500 bar to about 1000 bar.
20. A composition comprising the composition as claimed in claim 1 and tertiary butyl hydroquinone.
21. The composition as claimed in claim 20, wherein the composition of claim 1 is at a concentration ranging from about 50ppm to about 100ppm.
22. The composition as claimed in claim 20, wherein the tertiary butyl hydroquinone is at a concentration ranging from about 50ppm to about 100ppm.
23. An oil, fat or cosmetic preparation, comprising the composition of claim 1 or claim 20.
24. A method of preparing the oil, fat or cosmetic preparation as claimed in claim 23, wherein said method comprises act of mixing the composition of claim 1 or claim 20 with the oil, fat or cosmetic ingredients.