FURANOCOUMARINS FREE BAKUCHIOL COMPOSITIONS AND THEIR METHOD OF PREPARATION THEREOF

Abstract

The present invention discloses a novel method of isolating pure bakuchiol, substantially free of furanocoumarins (psoralen and isopsoralen) from the seeds of Psoralea sp using a combination of processes that include supercritical fluid extraction, high vacuum distillation and column purification. The invention also disclose compositions comprising bakuchiol that is free of furanocoumarins.

Claims

1. A process of isolating pure bakuchiol (not less than 99% w/w) as represented in STR #1, substantially free of furanocoumarins from Psoralea sp., comprising steps of: a) Powdering Psoralea sp. plant parts and pulverizing using 1.5 mm mesh and passing through magnetic separator to obtain a coarse powder; b) Subjecting the enriched extract of step b) to super critical fluid extraction (SCFE) using liquid CO.sub.2 in the presence of an entrainer and a stabilizing agent to obtain two fractions: low volatile compoundsS1 fraction, high volatile compoundsS2 fraction and a spent material (residue); c) Identifying the compounds in low volatile and high volatile fractions of step b) as bakuchiol, wherein the total yield of bakuchiol in S1 fraction is 5-15% w/w and 0.5-3% w/w in S2 fraction; d) Pooling the S1 and S2 fractions from step b) to obtain bakuchiol enriched extract, substantially free of furanocoumarins; e) Subjecting the bakuchiol enriched extract of step d) to high vacuum distillation (HVD) at desired temperature with vacuum to obtain different bakuchiol enriched fractions and a pot residue, wherein the bakuchiol content is assayed using HPLC; f) Pooling the bakuchiol enriched fractions of step e) and passing them through a Silica gel column prepacked with heptane and eluting using a suitable mobile phase and collecting different fractions to get obtain content of bakuchiol (99% w/w) as assayed by HPLC; and g) Collecting the fractions with bakuchiol content of not less than 99% w/w and further subjecting the fractions to Thin Layer chromatography with suitable solvents as mobile phase with detection under wavelength UV 254 nm, to obtain bakuchiol free or substantially free of furanocoumarins ##STR00002##

2. The process as in claim 1, wherein the furanocoumarins are psoralen and isopsoralen.

3. The process as in claim 1, wherein the stabilizing agent in SCFE is selected from the group consisting of rosmarinic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherols.

4. The process as in claim 1, wherein the entrainer of step b) is selected from the group consisting of ethanol, methanol, isopropyl alcohol, acetone, chloroform, benzene, hexane, heptane, cyclohexane and combinations thereof.

5. The process as in claim 1, wherein the entrainer of step b) is selected from the group consisting of ethanol, methanol and isopropyl alcohol.

6. The process as in claim 1, wherein the mobile phase in step f) is selected from the group consisting of heptane and ethyl acetate or combinations thereof.

7. The process as in claim 1, wherein the mobile phase in step g) is selected from the group of n-heptane and ethyl acetate or combinations thereof, wherein the ratio of heptane and ethyl acetate is between 2-9:1-8.

8. A composition comprising bakuchiol free or substantially free of furanocoumarins, wherein the bakuchiol is isolated from Psoralea sp., using the process as mentioned in claim 1.

9. The composition as in claim 8, wherein the composition further comprises stabilizing agents, bioavailability enhancers and antioxidants, pharmaceutically or nutraceutically or cosmeceutically accepted excipients and enhancers and suitable for administration in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies, eatables, creams, lotions, ointments, serum, gels or spray.

Description

EXAMPLES

Example 1

Isolation of Bakuchiol Free of Furanocoumarins

[0053] The present inventions discloses a novel process for the isolation of bakuchiol

[0054] (99% w/w) free or substantially free of furanocoumarins (psoralen and isopsoralen). It is a three-step process containing the following:

[0055] STEP 1: Subjecting the Psoralea raw material to SCFE extraction under controlled conditions. The content of bakuchiol in Psoralea seed on raw material basis is 4.5-5.5% w/w by HPLC. After subjecting to SCFE the pooled SCFE extract contains 35-40% w/w assay by HPLC

[0056] STEP 2: The pooled SCFE extract is subjected to High vacuum distillation under controlled temperature and pressure conditions, wherein the assay of Bakuchiol is enriched from 40% to 66% w/w by HPLC.

[0057] STEP 3: Isolating and purifying bakuchiol free or substantially free of furanocoumarins by Silica gel column chromatography. Bakuchiol 66% w/w is purified/passed through Silica gel column chromatography, to obtain pure bakuchiol 99% w/w by HPLC.

[0058] The above steps are disclosed in detail below:

[0059] The Psoralea raw material is initially extracted with SCFE along with an entrainer to obtain an enriched extract, wherein the entrainer is selected form methanol, ethanol or isopropyl alcohol.

[0060] The below flow chart explains the SCFE extraction of bakuchiol form Psoralea corylifolia seeds (step 1)

[0061] The content of Bakuchiol and Furanocoumarins is proportionately high in the S1 fraction. SCPE extraction is performed with the above condition and the cumulative results in each hour is described there in the below table1.

TABLE-US-00001 TABLE 1 SCFE extraction S1 FRACTION S2 FRACTION SCFE extraction Extraction time Extraction time Extraction time 0-1 hour 0-2 hour 0-5 hour Input (100 Kg) 55.1 Kg 55.3 Kg 51 Kg 55 Kg Description of Yellowish Yellowish Yellowish Dark brownish the output brown coloured brown coloured brown coloured oil viscous oil with viscous oil with viscous oil with characteristic characteristic characteristic odour odour odour Assay 44.23% 43.28% 39.88% 24.63% Bakuchiol by HPLC % w/w Psoralen 2.63% 3.06% 3.9% 42.69%

[0062] From the above table it is evident that S1 fraction has the highest assay of Bakuchiol (between 39.88% to 44.23%) whereas in S2 fraction Bakuchiol content becomes proportionately reduced to 24.63%. However, the Psoralen content in S2 fraction is higher (42.69%).

[0063] After SCFE extraction, the fractions were subjected to High vacuum distillation (HVD) at desired temperature with controlled vacuum (0.5 mm Hg) to obtain a bakuchiol extract free of psoralen and isopsoralen. The mechanism behind the distillation is probably to open the lactone ring of the furanocoumarin. Table 2 discloses the results of HVD.

TABLE-US-00002 TABLE 2 High vacuum distillation of Psoralea corylifolia Distillation Input Quantity: 55.1 Kg (Assay: 44.23% by HPLC) Still Vapour Bakuchiol Fraction Temp Temp Vacuum Output Assay Number ( C.) ( C.) (mm Hg) (kg) (% w/w) Fraction 1 195-233 150-192 0.5 6.06 66.02 Fraction 2 233-250 192-197 0.5 4.35 61.70 Fraction 3 250-265 198-205 0.5 1.65 78.13 Bottom mass 40.22 2.70 (pot residue)

[0064] The results indicate that Fractions 1,2 and 3 are enriched with bakuchiol and thus all the three fractions can be pooled together. The pot residue after pooling the fractions may generally be enriched with fatty acids with less bakuchiol and thus is discarded

[0065] Finally, the fractions from the HVD (from lower assay of bakuchiol 12.06 Kg, 66.15% average assay), are passed through Silica gel column chromatography which is prepacked with heptane and fractions are collected from 100% heptane to 2.0% Ethyl acetate/heptane. Similar fractions were pooled to get higher content of pure bakuchiol (6.69 Kg, 99% w/w) (Table 3). Analysis is performed by high performance liquid chromatography (HPLC); Fraction details as given below in Table3:

TABLE-US-00003 TABLE 3 Silica gel column purification Content of Fractions Bakuchiol Fractions quantity % (w/w) Number Eluted Solvent (kg) by HPLC 1 to 11 Heptane only 0.12 70.18% 12 to 16 0.5% (v/v) Ethyl acetate/heptane 0.72 83.77% 17 1% (v/v) Ethyl acetate/heptane 0.89 99.05% 18 1% (v/v) Ethyl acetate/heptane 0.95 19 1% (v/v) Ethyl acetate/heptane 0.78 99.12% 20 1% (v/v) Ethyl acetate/heptane 0.61 21 1% (v/v) Ethyl acetate/heptane 1.34 99.26% 22 1% (v/v) Ethyl acetate/heptane 1.31 99.50% 23 1% (v/v) Ethyl acetate/heptane 0.25 99.63% 24 2% (v/v) Ethyl acetate/heptane 0.56 99.75% 25 2% (v/v) Ethyl acetate/heptane 0.56 58.12% Total quantity 6.69 kg 99%

[0066] From the above fractions, fraction No. 17-24 (6.69 Kg of pure 99% bakuchiol) were collected separately by further purification and monitored using Thin layer chromatography [Mobile phase=Heptane: Ethyl acetate (7:3)] detection under UV wavelength 254 nm to yield a bakuchiol extract that is substantially free of furanocoumarins.

Example 2

Formulations

[0067] Tables 4-7 provide illustrative examples of nutraceutical and cosmeceutical formulations

TABLE-US-00004 TABLE 4 Tablet Active Ingredients Bakuchiol (Babchiol) Excipients Microcrystalline cellulose, Colloidal silicon dioxide, Magnesium stearate, BioPerine, Polyvinylpyrrolidone/starch/Hydroxy propyl methyl cellulose, Hydroxy propyl cellulose

TABLE-US-00005 TABLE 5 Capsule Active Ingredients Bakuchiol (Babchiol) Excipients Microcrystalline cellulose, Colloidal silicon dioxide, Magnesium stearate, BioPerine, Polyvinylpyrrolidone/starch/Hydroxy propyl methyl cellulose, Hydroxy propyl cellulose

TABLE-US-00006 TABLE 6 Powder Active Ingredients Bakuchiol (Babchiol) Excipients BioPerine, Starch

TABLE-US-00007 TABLE 7 Cream Phase Ingredients Functions A Purified Water Diluent Disodium EDTA Chelating Agent Biopol Crystals Thickener Glycerin Humectant B Olivem 1000 Emulsifier DUB CO Emollient GMS SE Emulsifier Cetyl Alcohol Emollient Shea Butter Emollient Imex IN3 Emollient Tinogard TT Anti Oxidant Tinogard TS Anti Oxidant C Tween 20 Solubilizer Tetrahydrocurcuminoids Skin Lightening/Whitening D Purified Water Diluent Bakuchiol (Babchiol) Skin Lightening/Whitening E Euxyl PE 9010 Preservative Xiameter PMX 3031 Detackification F Purified Water Diluent Sodium Metabisulfite Anti Oxidant

[0068] While the invention has been described with reference to a preferred embodiment, it is to be clearly understood by those skilled in the art that the invention is not limited thereto. Rather, the scope of the invention is to be interpreted only in conjunction with the above embodiments.