1. Supercritical Carbon Dioxide Extraction of Cocoa Butter Analogy Fat from Mango Seed Kernel Oil: An Optimization Approach of Response Surface Methodology PPTI, USM: Md. J.H. Akanda, Prof. A.K. Md. Omar, Prof. Nik Norulaini, M. M. Bennama, S. Ferdosh Faculty of Pharmacy, IIUM: Prof. I.S.M. Zaidul, K. M. Sharif, J. Azmir Food Science, UPM: Prof. J. Selamat Presenter Mohammad Sharif Khan

2. Introduction Mango (Mangifera Indica L.) is a popular commercial tropical fruit across the globe. In Malaysia, mango fruit is very popular. The juice is very common and easily available as there are many mango juice industries in the country. Between 1971 and 1993, the production of mango worldwide increased by nearly 50% (Galan, 1993).

3. Introduction Cont. (Source: FAO, 1992 - 2010) Production by region

4. Introduction The major mango growing countries are China, Mexico, Pakistan, Bangladesh, India, Indonesia, Thailand, Philippines, Malaysia, Brazil, Australia, Nigeria and Egypt. In this industry, only the pulp of mango is used for the production of juice, syrup, jam, jelly, etc. In mango juice industries, mango seed kernels (MSK) are the major by-products (Ajila et at., 2007).

5. Introduction Cont.. Huge amount of mango seeds are discarded as agricultural waste, creating both disposal and pollution problems (Nuraini et al., 2008). The MSKs represents up to 25% of the fruits, which are left as waste after the consumption of the fruit, contain about 7.1-15% of total fat (from the kernel of the fruit). The fat extracted from the MSKs has a yellow color and melts at 35 to 38 ºC.

6. Introduction Cont.

7. These fats received considerable attention for their suitable physical properties for fabricating of confectionary. Ali et al., (1985) and Hemavathy et al., (1987) investigated the fatty acid constituents of MSKs to be used as cocoa-butter substitute in confectionery industry (Solvent extraction). Compositional studies of the MSKs have been carried out by many researchers. To this date, no studies have been conducted on the use of CO2 as solvent for the extraction and fractionation of oil from MSK. Introduction Cont.

8. SFE- powerful technique in separation process where CO 2 employed as a supercritical fluid CO 2 – Non-toxic, non-flammable, relatively low cost solvent highly soluble with lipid / Oil Pressure Temperature Triple Point Gas Supercritical Fluid Region Critical Point Solid Liquid 7.28 MPa, 31.1ºC Phase Diagram of a Pure Components They have a combination of vapor and liquid properties. They have densities and viscosities less than liquids. Their diffusivities are gas-like. They transfer mass very rapidly. They are compressible and homogeneous.

9. ADVANTAGES OF SC-CO 2 Environmentally Friendly Quick Extraction Low Operating Temperature High Quality Oil Cost Effective Advantages of Supercritical Carbon Dioxide Extraction

10. Objectives To optimize the extraction parameters namely: pressure, temperature and CO 2 flow rate using RSM for SC-CO 2 extraction of MSK oil from MSK to achieve the highest yield as well as compaire with Soxhlet method.

11. Materials and Methods Soxhlet extraction Supercritical CO2 extraction -MSKs -Hexane -Carbon dioxide -Sodium methoxide solution

12. Result and Discussions

13. Optimization the extraction condition of SC-CO 2 Optimization MSK oil extraction GC analysis (3 replications)

14. Experimental Design  Optimize the extraction condition of oil yield from the MSK was carried out using RSM  A central composite design consist of 20 experimental runs with six replication at central points were employed to fit the model  The designs variables were the pressure, temperature, and flow rate while response variable was oil yield

15. Optimization Design Independent variables SymbolsVariable levels 0+1 Pressure (MPa) X1X1 203550 Temperature (º C) X2X2 406080 CO 2 flow rate (mL/min) X3X3 12.54 Full factorial central composition (CCD) design of RSM with 3 factors has been used to optimize for the MSK oil

16. Design matrix, experimental and predicted values in the screening design. Run numberIndependent variables Oil yield (%) Pressure, X 1 (MPa) Temperature X 2 (ºC) Flow rate X 3 (ml/min) Experimental (%) Predicted (%) 120602.53.053.40 2356014.194.68 335402.56.286.72 435802.58.227.80 53560410.259.78 650602.511.1310.80 735602.59.119.02 835602.59.09.02 935602.59.099.12 1025.847.71.63.412.98 1125.872.21.63.212.95 1225.872.23.45.345.65 1335602.59.109.12 1435602.59.149.12 1544.247.83.49.719.95 1644.272.31.69.539.73 1725.847.73.47.897.67 1844.272.23.410.8711.29 1944.247.71.66.736.40 2035602.59.109.12

17. Estimated regreesion co-efficient and P- value of MSK oil for the SC-CO2 extraction Term Coef SE Coef T P Constant -30.5628 5.53515 -5.522 0.000 Block 0.0531 0.10379 0.512 0.621 Pressure 0.4798 0.14349 3.344 0.009 Temperature 0.4033 0.11793 3.420 0.008 Flow rate 9.5154 1.32240 7.196 0.000 Pressure*Pressure -0.0085 0.00148 -5.739 0.000 Temperature*Temperature -0.0044 0.00083 -5.260 0.001 Flow rate*Flow rate -0.7935 0.14833 -5.350 0.000 Pressure*Temperature 0.0075 0.00143 5.216 0.001 Pressure*Flow rate -0.0339 0.01906 -1.780 0.109 Temperature*Flow rate -0.0443 0.01429 -3.102 0.013 P-value ≤0.05 indicates that statistically significant

18. TermCoefficientsStandard error tp-value βoβo -30.56285.53515-5.5220.000 X 1 (β 1 )0.47980.143493.3440.009 X 2 (β 2 )0.40330.117933.4200.008 X 3 (β 3 )9.51541.322407.1960.000 X 1 2 (β 11 )-0.00850.00148-5.7390.000 X 2 2 (β 22 )-0.00440.00083-5.2600.001 X 3 2 (β 33 )-0.79350.14833-5.3500.000 X 1 *X 2 (β 12 )0.00750.001435.2160.001 X 1 *X 3 (β 13 )-0.03390.01906-1.7800.109 X 2 *X 3 (β 23 )-0.04430.01429-3.1020.013 98.55 Adj.96.95 All the linear and quadratic effect and their interaction between variables exhibited significantly (p<0.05) positive effect on the MSK oil yields except X 1 *X 3 Estimated regreesion co-efficient and P- value of MSK oil for the SC-CO2 extraction

19. Effect of temp. & pres. at constant CO2 flow rate on yield of MSK The pressure had greatly positive effect on MSK oil yield due to the increase of solvent power of supercritical CO2 resulting from the increment of density of CO2

20. Pressure and the CO2 flow rate both exhibited significantly positive effect on the total MSK oil yield. Effect of pres. & flow rate on yield of MSK at cons. Tem.

21. Effect of temp & flow rate on yield of MSK at cons. Pres. The flow rate was increased from 1 to 2.5 ml/min, there was a sharp increase of MSK oil yield from 4.19 to 9.14%

22. Conclusions The general availability of the model was determined using coefficient of determination (R 2 ) optimal condition for MSK oil yield was found to be at 44.2 MPa, 72.2 ºC and CO2 flow rate at 3.4 ml/min. The MSK oil yield by SFE was close to the yield of soxhlet method 11.29% by SFE and 11.7% by Soxhlet extraction method

23. References Ajila, C.M., Naidu, K.A., Bhat, S.G., Prasada Rao, U.J.S. (2007). Bioactive compounds and antioxidant potential of mango peel extract. Food Chemistry, 105, 982-988. Galan SV (1993) The situation of mango culture in the world. Acta Hort 341:31–38. Food And Agricultural Organization of United Nations – FAOSTAT. (2010). http://faostat3.fao.org/home/index.htmlhttp://faostat3.fao.org/home/index.html Nuraini, J., Norziah, M.H., Tagally, B.Z., Lim, S.F., Norita, M., Fazilah, A., (2008). Extraction of fish oil from fish waste from surimi processing plant. Int. Conf. Environ. Res. Technol. 28–30 May, Penang, Malaysia, 144– 148. Ali, M.A., Gafur, M.A., Rahman, M.S., Ahmed, G.M. (1985). Variations in fat content and lipid class composition in ten different varieties. Journal of the American Oil Chemists’ Society, 62(3), 520-523. Hemavathy, J., J.V. Prabhakar, D.P. Sen, 1987. Composition of polar lipids of alphonso mango (Mangifera indica) kernel. Journal of Food Science, 52(3), 833-834.

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