Food & Feed Research


Volume 45, Issue 1
vitamin C, lipase, ascorbyl linoleate, antioxidant, diffusivity
TOOLS Creative Commons License
Marija M. Ćorović*1, Ana D. Milivojević2, Milica B. Carević1, Katarina M. Banjanac2,
Ljubodrag V. Vujisić3, Rada V. Pjanović1, Dejan I. Bezbradica1
1University of Belgrade, Faculty of Technology and Metallurgy, 11000 Belgrade, Karnegijeva 4, Serbia
2University of Belgrade, Faculty of Technology and Metallurgy, Innovation Center, 11000 Belgrade, Karnegijeva 4, Serbia
3University of Belgrade, Faculty of Chemistry, 11000 Belgrade, Studentski trg 12, Serbia


Lipophilic derivatives of vitamin C are additives with antioxidant properties, attractive for application in food, cosmetics and pharmaceutics. They could be synthesized in lipase-catalyzed processes by using various acyl donors. Hereby, we present application of linoleic acid, which is polyunsaturated fatty acid essential in human nutrition, for esterification of vitamin C catalyzed by immobilized enzyme preparation Novozym® 435 in acetone. Highest specific ester yield, 9.7 mmol/g of immobilized lipase, was accomplished with 0.15 M of vitamin C, 0.6 M of linoleic acid, 3 g/l of enzyme and 0.07% (v/v) of water, at 60 °C. NMR analyses of purified product proved that synthesized molecule was identical to 6-O-ascorbyl linoleate. Capacity of ester for scavenging 2,2-diphenyl-1-picrylhydrazyl radicals was two times higher comparing to parent molecule, vitamin C. Its diffusion coefficient, determined using Franz cell and cellulose acetate membrane, was 40% higher than palmitate and 62% higher than oleate. Obtained results showed that L-ascorbyl linoleate could be successfully synthesized in biocatalyzed processes. Furthermore, it was demonstrated that it possess high potential for application in different lipophilic products due to its liposolubility, high antioxidant efficiency and good diffusion properties.

Download full article PDF


  1. Adamczak, M., Bornscheuer, U.T. (2009). Improving ascorbyl oleate synthesis catalyzed by Candida antarctica lipase B in ionic liquids and water activity control by salt hydrates. Process Biochemistry, 44 (3), 257-261.
  2. Andersen, F.A. (1999). Final report on the safety assessment of ascorbyl palmitate, ascorbyl dipalmitate, ascorbyl stearate, erythorbic acid, and sodium erythorbate. International Journal of Toxicology, 18 (SUPPL. 3), 1-26.
  3. Ando, H., Shimizu, M., Hashimoto, A., Kato, H., Ozasa, Y. (1992). Skin whitening cosmetics. U.S. Patent No. 5,078,989. U.S. Patent and Trademark Office, Washington, DC.
  4. Austria, R., Semenzato, A., Bettero, A. (1997). Stability of vitamin C derivatives in solution and topical formulations. Journal of Pharmaceutical and Biomedical Analysis, 15 (6), 795-801.
  5. Bezbradica, D., Ćrovic, M., Tanasković, S. J., Luković, N., Carević, M., Milivojević, A., Knezević-Jugović, Z. (2017). Enzymatic syntheses of esters - Green chemistry for valuable food, fuel and fine chemicals. Current Organic Chemistry, 21 (2), 104-138.
  6. Bradoo, S., Saxena, R.K., Gupta, R. (1999). High yields of ascorbyl palmitate by thermostable lipase-mediated esterification. JAOCS, Journal of the American Oil Chemists' Society, 76 (11), 1291-1295.
  7. Burham, H., Rasheed, R.A. G.A., Noor, N. M., Badruddin, S., Sidek, H. (2009). Enzymatic synthesis of palm-based ascorbyl esters. Journal of Molecular Catalysis B: Enzymatic, 58 (1-4), 153-157.
  8. Chen, B., Guo, Z., Let, M. B., Lue, B.-M., Xu, X. (2008). Preparation of CLA ascorbyl ester with improved volumetric productivity by an ionic liquid-based reaction system. Organic and Biomolecular Chemistry, 6 (17), 3196-3201.
  9. Ćorović, M., Mihailović, M., Banjanac, K., Carević, M., Milivojević, A., Milosavić, N., Bezbradica, D. (2017). Immobilization of Candida antarctica lipase B onto Purolite® MN102 and its application in solvent-free and organic media esterification. Bioprocess and Biosystems Engineering, 40 (1), 23-34.
  10. Ćorović, M., Milivojević, A., Carević, M., Ba-njanac, K., Jakovetić Tanasković, S., Bezbradica, D. (2017). Batch and semicontinuous production of L-ascorbyl oleate catalyzed by CALB immobilized onto Purolite® MN102. Chemical Engineering Research and Design, 126, 161-171.
  11. Cousins, R.C., Seib, P.A., Hoseney, R.C., Deyoe, C.W., Liang, Y. T., Lillard, D. W. (1977). Synthesis of 6-fatty acid esters of l-ascorbic acid. Journal of the American Oil Chemists' Society, 54 (8), 308-312.
  12. Hsieh, H.J., Chen, J. W., Giridhar, R., Wu, W. T. (2005). Synthesis of mixed esters of ascorbic acid using methyl esters of palm and soybean oils. Preparative Biochemistry and Biotechnology, 35 (2), 113-118.
  13. Karmee, S.K. (2009). Biocatalytic synthesis of ascorbyl esters and their biotechnological applications. Applied Microbiology and Biotechnology, 81 (6), 1013-1022.
  14. Klimundová, J., Šatinský, D., Sklenářová, H., Solich, P. (2006). Automation of simultaneous release tests of two substances by sequential injection chromatography coupled with Franz cell. Talanta, 69 (3), 730-735.
  15. Liu, Y., Wang, J., Yan, Y., Li, J. (2011). Biocatalytic synthesis and antioxidant capacities of ascorbyl esters by Novozym 435 in tertbutanol system using different acyl donors. African Journal of Biotechnology, 10 (75), 17282-17290.
  16. Moreno-Perez, S., Filice, M., Guisan, J.M., Fernandez-Lorente, G. (2013). Synthesis of ascorbyl oleate by transesterification of olive oil with ascorbic acid in polar organic media catalyzed by immobilized lipases. Chemistry and Physics of Lipids, 174, 48-54.
  17. Nickels, H.D., Hackenberger, A.D. (1987). Process for producing fatty acid esters of ascorbic acid. U.S. Patent No. 4,705,869. U.S. Patent and Trademark Office, Washington, DC.
  18. Park, S., Viklund, F., Hult, K., Kazlauskas, R. J. (2003). Vacuum-driven lipase-catalysed direct condensation of L-ascorbic acid and fatty acids in ionic liquids: Synthesis of a natural surface active antioxidant. Green Chemistry, 5 (6), 715-719.
  19. Pjanović, R., Bošković-Vragolović, N., Veljković Giga, J., Garić-Grulović, R., Pejanović, S., Bugarski, B. (2010). Diffusion of drugs from hydrogels and liposomes as drug carriers. Journal of Chemical Technology and Biotechnology, 85 (5), 693-698.
  20. Salis, A., Pinna, M., Monduzzi, M., Solinas, V. (2005). Biodiesel production from triolein and short chain alcohols through biocatalysis. Journal of Biotechnology, 119 (3), 291-299.
  21. Sharma, O.P., Bhat, T.K. (2009). DPPH antioxidant assay revisited. Food Chemistry, 113 (4), 1202-1205.
  22. Song, Q.-X., Wei, D.-Z., Zhou, W.-Y., Xu, W.-Q., Yang, S.-L. (2004). Enzymatic synthesis and antioxidant properties of L-ascorbyl oleate and L-ascorbyl linoleate. Biotechnology Letters, 26 (23), 1777-1780.
  23. Song, Q., Zhao, Y., Xu, W., Zhou, W., Wei, D. (2006). Enzymatic synthesis of L-ascorbyl linoleate in organic media. Bioprocess and Biosystems Engineering, 28 (4), 211-215.
  24. Song, Q. X., Wei, D. Z. (2002). Study of Vitamin C ester synthesis by immobilized lipase from Candida sp. Journal of Molecular Catalysis B: Enzymatic, 18 (4-6), 261-266.
  25. Špiclin, P., Gašperlin, M., Kmetec, V. (2001). Stability of ascorbyl palmitate in topical microemulsions. International Journal of Pharmaceutics, 222 (2), 271-279.
  26. Stojanovic, M., Carevic, M., Mihailovic, M., Ve-ličkovic, D., Dimitrijevic, A., Milosavic, N., Bezbradica, D. (2015). Influence of fatty acid on lipase-catalyzed synthesis of ascorbyl esters and their free radical scavenging capacity. Biotechnology and Applied Biochemistry, 62 (4), 458-466.
  27. Stojanović, M., Veličković, D., Dimitrijević, A., Milosavić, N., Knežević-Jugović, Z., Bezbradica, D. (2013b). Lipase-catalyzed synthesis of ascorbyl oleate in acetone: Optimization of reaction conditions and lipase reusability. Journal of Oleo Science, 62 (8), 591-603.
  28. Stojanović, M. M., Carević, M. B., Mihailović, M. D., Knežević-Jugović, Z.D., Petrović, S.D., Bezbradica, D. I. (2013a). Enzymatic synthesis and application of fatty acid ascorbyl esters. Hemijska Industrija, 67 (2), 239-247.
  29. Tamalampudi, S., Talukder, M. R., Hama, S., Numata, T., Kondo, A., Fukuda, H. (2008).
  30. Enzymatic production of biodiesel from Jatropha oil: a comparative study of immobilized-whole cell and commercial lipases as a biocatalyst. Biochemical Engineering Journal, 39 (1), 185-189.