A PROCESS FOR EXTRACTION AND SEPARATION OF OXYRESVERATROL FROM ARTOCARPUS LAKOOCHA ROXB

Abstract

The present invention relates to a process for separation of oxyresveratrol molecule from the extracted solution of Artocarpus lakoocha Roxb. through membrane application. The product can be obtained in excellent yield upto 81% in case of extraction using water as solvent and can be separated from the extracted mixture upto 98% using indigenously developed nanofiltration membrane. Only the desired Trans isomer is obtained and no cis isomerisation takes place during the extraction process.

Claims

1) A process for extraction and separation of oxyresveratrol from the heartwood of Artocarpus lakoocha Roxb., said process comprising the steps of: (i) Mixing Artocarpus lakoocha Roxb. powder with water at a temperature in the range of 30° C.-70° C. for 14 to 10 hours and filtering to obtain a solution of the extract; (ii) subjecting the solution of the extract obtained in step (i) to membrane treatment in a two compartment membrane cell containing a nanofiltration membrane selected from Alpha cyclodextrin-polysulfone composite nanofiltration membrane, Beta cyclodextrin-polysulfone composite nanofiltration membrane or Gama cyclodextrin-polysulfone composite nanofiltration membrane by circulating through the said nanofiltration membrane by using a peristaltic pump at a flow rate in the range of 42.50 ml/min to 107.14 ml/min; and (iii) collecting the oxyresveratrol from the feed side of membrane.

2. The process as claimed in claim 1, wherein the mixing of Artocarpus lakoocha Roxb. powder with water is done at 50° C.

3. The process as claimed in claim 1, wherein the mixing of Artocarpus lakoocha Roxb. powder with water is done for 4 hours.

4. The process as claimed in claim 1, wherein the nanofiltration membrane is beta cyclodextrin-polysulfone composite nanofiltration membrane.

5. The process as claimed in claim 1, wherein two compartment membrane cell has a feed side and a permeate side with volumes of 150 ml and 120 ml respectively.

6. The process as claimed in claim 1, wherein the trans membrane pressure is in the range of 7.2 psi to 21.75 psi (49642.3 Pa to 149960.97 Pa), and optimum pressure being 14.50 psi (99973.98 Pa).

7. (canceled)

8. The process as claimed in claim 1, wherein the membrane used is reusable for more than ten cycles with consistent activity by backwashing the membrane with ethanol.

9. The process as claimed in claim 1, wherein the oxyresveratrol obtained is trans isomer of oxyresveratrol.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] FIG. 1: Flow diagram of permeation experiment the whereas the parts are as following [0032] 1. Membrane cell [0033] 2. Magnetic stirrer [0034] 3. Magnetic capsule [0035] 4. Membrane [0036] 5. Feed tank [0037] 6. Peristaltic pump [0038] 7. N.sub.2 gas [0039] 8. Gas valve [0040] 9. Gas valve [0041] 10. Water vessel [0042] 11. Sample collecting valve [0043] 12. Pressure gauze [0044] 13. Pressure gauze

[0045] FIG. 2: Experimental set up of separation experiment

DETAILED DESCRIPTION OF THE INVENTION

[0046] The biological material i.e. Artocarpus lakoocha Roxb. used for the process of the present invention is procured from Area: Kakojan P. O. Kakojan Pin code: 785107, Assam, India.

[0047] The present invention relates to process for the extraction and separation of oxyresveratrol from heartwood of Artocarpus lakoocha Roxb. using water as extracting solvent and membrane technology as separation technique. In the process, water is selected as the best extracting agent in comparison to the other solvents used. The said process comprises the mixing of Artocarpus lakoocha Roxb. powder with warm water at 50° C. for 4 hours, filtration of the powder through filter paper. For separation of oxyresveratrol from the extractant, the filtrate is subjected to membrane treatment in a two compartment membrane cell containing beta-cyclodextrine-polysulfone composite nanofiltration membrane for separation of the product. The membrane is reusable for several cycles with consistent activity by backwashing with ethanol.

[0048] The membranes used for the separation process is beta cyclodextrin composite with polysulfone. The polysolfone-beta cyclodextrin (PS-CD) composite membranes are prepared indigenously using phase inversion technique. The separation experiment is carried out in a disproportionate two-compartment membrane cell whose compartment volumes on the feed side and permeate side are 150 and 120 ml respectively. The polymeric membrane is placed between the compartments with silicone-rubber packing and the cell is connected with a reservoir of 500 ml capacity. The solutions of the extracted compound of concentration in the range 0.64 mmolL.sup.−1 to 0.76 mmolL.sup.1 in aqueous phase is stirred continuously and circulated through the membrane by using a peristaltic pump at the flow rate range of 42.50 ml/min to 107.14 ml/min. The area of the membrane is 19.6 cm.sup.2 and the trans membrane pressure of the experiment is in the range of 7.2 psi (49642.3 Pa) to 21.75 psi (149960.97 Pa). The sample solutions are collected at an interval of one hour for seven hours from the feed side and analyzed by UV and HPLC.

Examples

Example 1: Extraction of Oxyresveratrol (General Method)

[0049] 10 gm of heart wood of Artocarpus lakoocha Roxb. is stirred continuously with different solvent such as water, methanol, ethanol, aqueous H.sub.2SO.sub.4 and aqueous HCl at 30° C. to 70° C. for 10 hours. The samples are collected at a regular interval of time and filtered. The filtrate is analyzed for the presence of oxyresveratrol by UV visible spectrophotometer. The absorbance peak of oxyresveratrol is obtained at 329 nm. The concentration of oxyresveratrol is calculated using the calibration curve.

Example 2: Optimization of the Procedure

[0050] From the extraction method given in Example 1 following results have been obtained

TABLE-US-00001 TABLE 1 Extraction of oxyresveratrol at different time in water Ratio of wood and solvent: 1:15 Temperature: 50° C. Sl. No. Time (hour) % of extraction 1 1 17.79 2 2 53.70 3 3 71.43 4 4 81.49 5 5 79.70 6 6 66.60 7 7 43.89 8 8 32.90 9 9 22.57 10 10 13.09

TABLE-US-00002 TABLE 2 Extraction of oxyresveratrol in different solvents and temperatures Ratio of wood and solvent: 1:15 Time: 4 hours % of extraction Sl. No. Solvent 30° C. 50° C. 70° C. 1 H.sub.2O 38.65 81.49 44.93 2 Methanol 19.20 20.0 15.54 3 Ethanol 23.00 25.96 18.21 4 Sulphuric acid (aq.) 5.52 6.21 2.25 5 Hydrochloric acid (aq.) 6.25 8.00 3.29

TABLE-US-00003 TABLE 3 Extraction of oxyresveratrol at different ratio of wood and solvent Solvent: Water, Temperature: 50° C., Time: 4 hours Ratio of wood and Sl. No. solvent (w/w) % of extraction 1 1:30 30.85 2 1:25 28.25 2 1:20 42.45 3 1:15 81.49 4 1:10 42.55

TABLE-US-00004 TABLE 4 Separation of oxyresveratrol by membrane Concentration of feed = 0.76 mmolL.sup.−1, Pressure = 14.50 psi (99973.98 Pa) Flow rate Flux Membrane (ml/min) (mmol m.sup.−1h.sup.−1) % of recovery Alpha cyclodextrin- 107.14 8.81 88.86 polysulfone 75 8.17 83.42 composite NF 42.85 5.89 56.62 membrane Beta cyclodextrin- 107.14 9.88 98.07 polysulfone 75 9.84 97.64 composite NF 42.85 9.08 91.26 membrane Gama cyclodextrin- 107.14 5.01 50.10 polysulfone 75 4.59 41.12 composite NF 42.85 4.02 32.54 membrane

TABLE-US-00005 TABLE 5 Separation of oxyresveratrol by membrane Concentration of feed = 0.76 mmolL.sup.−1, Flow rate = 107.14 ml/min Pressure Pressure Flux % of Membrane (psi) (Pa) (mmol m.sup.−1h.sup.−1) recovery Alpha cyclodextrin- 7.20 49642.3 5.81 63.18 polysulfone 14.50 99973.98 8.81 88.86 composite NF 21.75 149960.97 9.02 90.69 membrane Beta cyclodextrin- 7.20 49642.3 5.81 64.43 polysulfone 14.50 99973.98 9.88 98.07 composite NF 21.75 149960.97 7.65 55.78 membrane Gama cyclodextrin- 7.20 49642.3 3.24 35.30 polysulfone 14.50 99973.98 5.01 50.10 composite NF 21.75 149960.97 4.23 42.25 membrane

TABLE-US-00006 TABLE 6 Separation of oxyresveratrol by membrane Pressure = 14.50 psi (99973.98 Pa), Flow rate = 107.14 ml/min Concentration Flux Membrane (mmolL.sup.−1) (mmol m.sup.−1h.sup.−1) % Recovery Alpha cyclodextrin- 0.76 5.89 88.86 polysulfone 0.69 5.21 56.28 composite NF 0.64 4.60 54.27 membrane Beta cyclodextrin- 0.76 9.88 98.07 polysulfone 0.69 8.78 92.56 composite NF 0.64 7.98 88.64 membrane Gama cyclodextrin- 0.76 5.01 50.01 polysulfone 0.69 3.21 43.65 composite NF 0.64 3.11 42.11 membrane

The Main Advantages of the Present Invention are:

[0051] 1. The method is very simple and environmentally benign. [0052] 2. The product can be obtained in excellent yield upto 81% in case of extraction using water as solvent. [0053] 3. The product can be separated from the extracted mixture upto 98% using indigenously developed nanofiltration membrane. [0054] 4. Only the desired trans isomer is obtained and no cis isomerisation takes place during the extraction process. [0055] 5. The oxyresveratrol molecule is easily recoverable from the membrane cell by backwashing with ethanol under a pressure limit within the NF range. [0056] 6. The process is continuous and recyclable. [0057] 7. The membrane can be reused for consecutively more than ten times for separation of oxyresveratrol without any loss in activity of the membrane. Also, there is no generation of any toxic chemical waste to the environment making the process simple, green, environmentally benign, economically viable and less energy intensive.

[0058] Other related references: [0059] 1. J. of Planar Chromatography 24(2011) 2,125-129 [0060] 2. Chem. Pharm. Bull. 58 (11) 1492 (2010) [0061] 3. Brajilian journal of Pharmacology, 123, 1691 (1998) [0062] 4. Brain Research, 1017, 98 (2004) [0063] 5. Journal of Biological Chemistry, 277(18), 16340 (2002) [0064] 6. US Patent No. 20040175788A1 (2004) [0065] 7. US 20120094361A1 (2012) [0066] 8. J. Applied Polymer Science, 125(5), 3388-3898, (2012) [0067] 9. CN101591680-B, Method for extracting oxidized resveratrol [0068] 10. CN101591680-A; CN101591680-B [0069] 11. Chemistry & Biodiversity, 2006, 3, 1138-1143 [0070] 12. Phytochemistry, 2012, 81, 42-49 [0071] 13. Pak. J. Pharm. Sci., 2010, 23(4), 403-408 [0072] 14. Journal of Natural Products, 2001, 64(11), 1457-1459 [0073] 15. Medicinal Principles and Practice, 2009: 18: 223-227