CLIAMS:We claim:
1. Treated Karanja oil having increased pongamol content of at least 2%.

2. Treated Karanja oil as claimed in claim 1, having pongamol content of 2 to 2.61%.

3. Treated Karanja oil having increased pongamol content of at least 2% obtained from Karanja oil having pongamol content of 0.2 to 0.9%.

4. Treated Karanja oil having pongamol content of 2 to 2.61% obtained from Karanja oil having pongamol content of 0.2 to 0.9%.

5. A process for the preparation of Karanja oil having increased pongamol content by treating raw Karanja oil with an organic acid, wherein the organic acid is concentrated acid, Karanja oil and organic acid are in the weight ratio 50 to 1000 % and the treatment is carried out at 50 to 120°C followed by removing the acid from the oil.

6. The process as claimed in claim 5 for the preparation of Karanja oil having pongamol content of at least 2%.

7. The process as claimed in claim 5 for the preparation of Karanja oil having pongamol content of 2 to 2.61%.

8. The process as claimed in claim 5, wherein the raw Karanja oil has a pongamol content of 0.2 to 0.9%.

9. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 50 to 200.

10. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 150 – 200%.

11. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 200%.

12. The process as claimed in claim 5, wherein the oil is treated with the acid at 60 to 100°C.

13. The process as claimed in claim 5, wherein the oil is treated with the acid at 75 - 85°C.

14. The process as claimed in claim 5, wherein the organic acid is selected from concentrated acetic acid, ascorbic acid, formic acid and lactic acid.

15. The process as claimed in claim 5, wherein the acid is removed from the oil by subjecting the oil to vacuum distillation followed by washing the oil with water and drying the oil.

16. The process as claimed in claim 15, wherein the oil is dried over anhydrous sodium sulphate.
,TagSPECI:FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
As amended by the Patents (Amendment) Act, 2005
&
The Patents Rules, 2003
As amended by the Patents (Amendment) Rules, 2014
COMPLETE SPECIFICATION
(See section 10 and rule 13)
TITLE OF THE INVENTION

Treated Karanja oil having increased pongamol content and process for the preparation thereof

APPLICANTS

Kancor Ingredients Ltd, an Indian Company, Rajan House, Appasabheb Marathe Marg, Prabhadevi, Mumbai 400 025, Maharashtra

INVENTORS
Vaikkathukattil Shaju Asokan, Kalappurakkal Ramesh Chandran, Vazhappilly Linsha Poulose and Unnikrishnan Prakash Kumar, all Indian Nationals and all of Kancor Ingredients Ltd, No VII/138, Kanakkankadavu Road, Angamaly South, Ernakulam – 683573

PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in which it is to be performed:

FIELD OF THE INVENTION
This invention relates to treated Karanja oil having increased pongamol content and process for the preparation thereof.

BACKGROUND OF THE INVENTION
Karanja oil is extracted from the Millettia pinnata seeds (Pongamia glabra seeds) by expeller or cold pressing or solvent extraction and is yellowish-orange to brown in colour. Besides the crystalline component named pongamol, Karanja oil also contains another crystalline compound named karanjin. Pongamol, commercially known as pongamia extract, has the chemical name 1-(4-methoxy-5-benzofuranyl)-3-phenyl 1,3 propanedione and the structural formula shown in Fig 1 of the accompanying drawings.

Karanja oil is styptic, antimicrobial and anthelmintic and is reported to be effective for treating diseases like leprosy, piles, ulcers, chronic fever, liver pain, rheumatism arthritis, scabies and whooping cough. (Warrier PK, Nambiar VPK, Ramankutty C.Indian Medicinal Plants. Vol.IV. Orient Longman Ltd., Madras 1995: 339p; Prasad G, Reshmi MV. A manual of medicinal trees. Agrobios India 132 p.a Propagation Methods. Foundation for Revitalization for Local Health Tradition, India 2003. CSIR. The Wealth of India: raw materials. Vol. 1-10. Council of Scientific and Industrial Research (CSIR), New Delhi, India 1948-98).

Karanja oil is highly recommended for inclusion in moisturizers and hair care products where emolliency is desired. In skin care, Karanja oil is used to treat eczema, psoriasis, skin ulcers and dandruff and to promote wound healing. WO 2014016349 A1 describes usage of Karanja oil for a photo protective composition. Madhavarao Buddepu et al reported that Karanja oil extract can be used to formulate highly effective sunscreen preparations as it enhances UV absorbing properties of sunscreens and helps in broadening the UV protection ability of the sunscreens. (Madhavarao Buddepu. Determination of In -Vitro Sunscreen Activity of Millettia pinnata (L.) Essential Oil. Drug Invention Today 01/2011; Vol 3:197-199). EP 0431755, DE 10140538 , WO 2009125182, EP 0521651 and US 5152983 reported pongamol as an ingredient for topical application to human skin and hair to provide enhanced protection from ultra violet light.

Recent research has revealed that pongamol is capable of inducing increase in glucose uptake in L6 myotubes, driven by a PI-3-K/AKT dependent mechanism. (Molecular and Cellular Endocrinology, 2011, 339(1-2), 90-104). It also reported to have antihyperglycemic activity (J. Ethnopharmacol. 2008, 118, 435-439). It has been used as an anticonvulsant CNS depressant (Ind. J. Pharma. Sci. 1994, 56, 163-167) and as potent intestinal alpha-glucosidase inhibitor (Bioorg Med Chem. 2009, 17(14), 5170-5175).

Different methods are reported for isolation of pongamol from Karanja oil. N.V.Bringi teaches a process for isolation of pongamol by molecular distillation of Karanja oil using high vacuum (J. Oil Tech. Ass. 1976, 8(3), 81). In another process Karanja oil is stored under refrigeration to remove precipitated karanjin and then it is extracted with alcohol to isolate pongamol. Alcohol extract of Karanja oil is stored and the precipitated material is taken in ether and crystallized to get pongamol (Ind J Pharm 1941, 3, 3 and Proc. of Ind. Acad. Sci 1942, 15A, 437).

R N Khana and T R Seshadri isolated pongamol from the alcohol extract using column chromatography with alumina as stationary phase and light petroleum as eluant. Pongamol was isolated from the light petroleum by counter current distribution between aqueous acetic acid and light petroleum. (Tetrahedron 1963, 19, 223). In another improvement the chromatographic step in the isolation of pongamol from the alcohol extract was eliminated and instead the isolation of pongamol from the alcohol extract was carried out by repeated partitioning of karanjin removed alcohol extract of Karanja oil between light petroleum and aqueous acetic acid. (R Aneja, R N Khana and T R Seshadri (J. Chem. Soc., 1963, p. 163)).

In another process for extraction of pongamol from Karanja oil, dry hydrogen chloride is passed through the alcohol extract of Karanja oil, and the precipitate separated is fractionally crystallized (J. Ind. Chem. Soc, Industrial and News Edition, 1954, 17, 39). GB2237805 describes extraction of pongamol from Karanja oil with organic acid followed by crystallization from solvents. Sangli variety (a region in Maharashtra State, India) Karanja oil (2000 g) was extracted with aqueous acetic acid (800 ml diluted with 200 ml water) at ambient temperature ( 25 - 30°C). Extract was aged at ambient temperature (25 - 30°C) for 3 days to obtain pongamol in 0.9% yield. The oil after isolation of pongamol is reported to be used for making soap. GB2237805 teaches extraction of Karanja oil with aqueous acid (v/v ratio 80 : 20) in the w/v ratio 20:1 at ambient temperature followed by ageing at ambient temperature. Pongamol yield reported is 0.9%.

Parmar et al reported 0.85% pongamol content in Karanja oil while V.K. Gore et al reported 0.9% pongamol content in Karanja oil. (Parmar BS, Shrawat KL and Mukerjee SK, Pongamia glabra: constituents and uses, J Sci Ind Res, 1976, 35, 608-611; V.K. Gore, P. Satyamoorthy. Determination of Pongamol and Karanjin in Karanja Oil By Reverse Phase HPLC. Analytical Letters.Vol. 33, Iss. 2, 2000). It is reported that different Karanja oil samples by HPLC analysis gave pongamol in the range of 0.85 to 0.9%. (Analytical Letters, 2000, 33(2), 337-346). However, chemical analysis of commercially available Sangli variety Karanja oil in our own laboratory showed a varying pongamol content of ~0.2-0.9%.

Extensive research and development activities are being carried out in the field of natural products, especially for use in pharmaceutical and cosmetic and dermatogical industries because of their being comparatively safe and cheap and naturally available. Therefore, there is need for natural products like Karanja oil having increased pongamol content, pongamol in high yield from Karanja oil and Karanja oil extract having high pongamol content and processes for the preparation thereof for various uses, especially pharmaceutical and cosmetic and dermatogical applications.

DESCRIPTION OF THE INVENTION
According to the invention there is provided treated Karanja oil having increased pongamol content of at least 2%, preferably 2 to 2.61%.

According to the invention there is also provided treated Karanja oil having increased pongamol content of at least 2%, preferably 2 to 2.61% obtained from Karanja oil having pongamol content of 0.2 to 0.9%.

According to the invention there is also provided a process for the preparation of Karanja oil having increased pongamol content by treating raw Karanja oil with an organic acid, wherein the organic acid is concentrated acid, Karanja oil and organic acid are in the weight ratio 50 to 1000 % and the treatment is carried out at 50 to 120°C followed by removing the acid from the oil.

Preferably, the oil is treated with the acid in the weight ratio 50 to 200%, still preferably in the weight ratio 150 – 200% and most preferably in the weight ratio 200%. Preferably, the oil is treated with the acid at 60 to 100°C and still preferably at 75 - 85°C. Preferably, the organic acid is selected from concentrated acetic acid, ascorbic acid, formic acid and lactic acid. However, other concentrated organic acids also can be used in the acid treatment.

In one variation of the process of the invention, the acid is removed from the oil by subjecting the oil to vacuum distillation followed by washing the oil with water and drying the oil. Preferably, the oil is dried over anhydrous sodium sulphate. The acid may be removed from the oil using other known procedures also.

According to the invention there is provided pongamol enriched Karanja oil which can be used inter alia in the pharmaceutical industry for therapeutic purposes. In cosmetic and dermatological industry, it can be used as a UVA absorber. Increased amount of the pongamol content increases the activity of the oil. Because of the increased activity, amount of oil required is reduced to give cost benefits. However, the treated oil can be used for other purposes also. The invention also provides a process for treating the oil and increasing and enriching the pongamol content thereof, which is simple to carry out and is cost effective.

The following examples are illustrative of the invention but not limitative of the scope thereof.

Example 1
Sangli variety Karanja oil having 0.6 % pongamol content was treated with glacial acetic acid in different proportions at 75 - 85°C for about 1 hr. Pongamol content of the oil after the acid treatment were as shown in the following Table 1:
Table 1
Oil % glacial acetic acid % pongamol %
3 g 10 % (0.3 g) 0.71
3 g 50 % (1.5 g) 2.06
3 g 100 % (3 g) 2.45
3 g 150 % (4.5 g) 2.48
3 g 200 % (6 g) 2.53
3 g 1000 % (30 g) 2.52

Table 1 shows that the pongamol content of the oil increased substantially with acid treatment of the oil in the weight percentages 50 to 1000 of the acid thereby indicating conversion of the various other unknown compounds in the oil into pongamol under the selective process parameters and conditions of the invention. Table 1 also shows that conversion of the various other unknown compounds in the oil into pongamol reaches saturation with acid percentage of 1000.

Example 2
Sangli variety Karanja oil (200g) having 0.6% pongamol content was treated with 400 g (50%) glacial acetic acid at 75 - 85°C for about 1 hr. Treated oil was concentrated by vacuum distillation, washed twice with 100 g water (1:0.5) and dried by passing through anhydrous sodium sulphate to get Karanja oil enriched with 2.61% pongamol content.

Example 3
Physico-chemical properties of the treated and untreated Karanja oil of Example 2 as found out by chemical analysis were as shown in the following Table 2:
Table 2
Properties Untreated Karanja oil Treated Karanja oil
Moisture percent by weight 0.25 0.10
Saponification value 190 185
Acid value 5.20 7.12
Color (Visual) Yellowish red Yellowish red
Odour (Olfactory) Nutty Nutty

Table 2 shows that there were no significant differences in the physico-chemical properties of both the treated and untreated oils.

Example 4
UVA protection factor of treated and untreated Karanja oil of Example 2 was evaluated using standard evaluation parameters commonly employed namely Boot Star Rating, PA (Protection Activity) Rating and Critical Wavelength (?c) measurement as follows:

Samples of treated and untreated Karanja oils of Example 2 were applied evenly in dots in light strokes at an application rate of 0.75mg/cm2 across the entire surface of PMMA (polymethyl methacrylate) plates. Samples were allowed to dry for 15 mins and exposed to Xenon flash lamp. Various parameters of the samples were measured with UV-2000S Ultraviolet Transmittance Analyzer Software and the results were as shown in the following Table 3 :
Table 3
Measurement Parameters Untreated Karanja oil Treated Karanja oil
UVA/UVB Ratio 0.476 0.753
Critical Wavelength 363.89 374.50
Boots Star Rating No Rating ***
UVA PF (Protection Factor) Nil 3.2
PA Rating No Rating PA+

Table 2 clearly shows that treated Karanja oil has Boot Star Rating and PA Rating while the untreated Karanja oil shows no rating and hence no UVA absorbance capability. Critical wavelength measurement shows that the treated oil has broader spectrum protection than the untreated oil. Untreated oil has no UVA protection factor, whereas treated oil has a protection factor of 3.2.

Example 5
HPLC (High Pressure Liquid Chromatography) profiles (chromatograms) of the untreated and treated Karanja oils of Example 2 were obtained and are as shown in Figs 2 and 3 of the accompanying drawings, respectively. It is quite clear from Figs 2 and 3 that the peak P1 representing pongamol in Fig 3 increased substantially as compared to the peak P representing pongamol in Fig 2 on treatment of the oil according to the invention thereby validating enrichment of the oil with pongamol content and the findings in Examples 1, 2 and 4.

We claim:
1. Treated Karanja oil having increased pongamol content of at least 2%.

2. Treated Karanja oil as claimed in claim 1, having pongamol content of 2 to 2.61%.

3. Treated Karanja oil having increased pongamol content of at least 2% obtained from Karanja oil having pongamol content of 0.2 to 0.9%.

4. Treated Karanja oil having pongamol content of 2 to 2.61% obtained from Karanja oil having pongamol content of 0.2 to 0.9%.

5. A process for the preparation of Karanja oil having increased pongamol content by treating raw Karanja oil with an organic acid, wherein the organic acid is concentrated acid, Karanja oil and organic acid are in the weight ratio 50 to 1000 % and the treatment is carried out at 50 to 120°C followed by removing the acid from the oil.

6. The process as claimed in claim 5 for the preparation of Karanja oil having pongamol content of at least 2%.

7. The process as claimed in claim 5 for the preparation of Karanja oil having pongamol content of 2 to 2.61%.

8. The process as claimed in claim 5, wherein the raw Karanja oil has a pongamol content of 0.2 to 0.9%.

9. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 50 to 200.

10. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 150 – 200%.

11. The process as claimed in claim 5, wherein the oil is treated with the acid in the weight ratio 200%.

12. The process as claimed in claim 5, wherein the oil is treated with the acid at 60 to 100°C.

13. The process as claimed in claim 5, wherein the oil is treated with the acid at 75 - 85°C.

14. The process as claimed in claim 5, wherein the organic acid is selected from concentrated acetic acid, ascorbic acid, formic acid and lactic acid.

15. The process as claimed in claim 5, wherein the acid is removed from the oil by subjecting the oil to vacuum distillation followed by washing the oil with water and drying the oil.

16. The process as claimed in claim 15, wherein the oil is dried over anhydrous sodium sulphate.
Dated this 24th day of September 2014

(Jose M A)
of Khaitan & Co
Agent for the Applicants
Reg No IN/PA-44