CUPRAC assay-guided profiling of antioxidant compounds in methanol extract of Lentinus squarrosulus Mont. mycelium

Sumaiyah ABDULLAH, Noorlidah ABDULLAH, Vikineswary SABARATNAM, Ken Choy YAP


A cupric reducing antioxidant capacity (CUPRAC)-guided purification approach was performed on a methanol extract of Lentinus squarrosulus (LsqMeOH) by using reversed phase-high performance liquid chromatography. Using reversed phase-high performance liquid chromatography, three fractions were separated arbitrarily named FR1, FR2 and FR3. Results showed that FR2 exhibited the highest antioxidant activity in CUPRAC assay (A450, 0.86) but not significantly different from LsqMeOH (A450, 0.84). FR1 and FR3 showed much lower absorbance, with values (A450, 0.21) and (A450, 0.36) respectively at 1 mg ml-1. The most active fraction (F3) was further subjected to LC-MS/MS to obtain its detailed chemical profile. Uridine, ganoderic acid derivative, and flavonoids were the first time being found in L. squarrosulus antioxidative fractions. The present results indicate that the fraction extracts of L. squarrosulus possess antioxidant properties and can be used as free radical inhibitors. Therefore, this research suggested the potentials of L. squarrosulus as a source of antioxidant extract to be used in food industries (functional food).


medicinal mushroom; active compounds; Lentinus squarrosulus

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Apak, R., Güclü, K., Ӧzyürek, M. & Karademir, S. E. (2004). Novel total antioxidant capacity index for dietary polyphenols and vitamin C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of Agriculture and Food Chemistry, 52, 7970-7981. doi:10.1021/jf048741x

Bae, A. H., Lee, S.W., Ikeda, M., Sano, M., Shinkai, S., Sakurai, K. (2004). Rod-like architecture and helicity of the poly(C)/schizophyllan complex observed by AFM and SEM. Carbohydrate Research, 339, 251-258. doi:10.1016/j.carres.2003.09.032

Barros, L., Calhelha, R. C., Vaz, J. A., Ferreira, I. C. F. R., Baptista, P. & Estevinho, L. M. (2007). Antimicrobial activity and bioactive compounds of Portuguese wild edible mushrooms methanolic extracts. European Food Research and Technology, 225, 151-156. doi:10.1007/s00217-006-0394-x

Barros, L., Duenas, M., Ferreira, I. C. F. R., Baptista, P., Santos-Buelga, C. (2009). Phenolic acids determination by HPLC-DAD-ESI/MS in sixteen different Portuguese wild mushrooms species. Food and Chemical Toxicology, 47, 1076-1079. doi:10.1016/j.fct.2009.01.039

Berg, J. M., Tymoczko, J. L. & Stryer L. (2002). Biochemistry (5th ed.). New York, NY: W H Freeman.

Bolhassan, M. H., Abdullah, N., Sabaratnam, V., Tsutomu, H., Abdullah, S., Rashid, N. M. N. & Musa, M. Y. (2012). Diversity and distribution of Polyporales in Peninsular Malaysia. Sains Malaysiana, 41, 155-161.

Chang, S. T. & Buswell, J. A. (2003). Medicinal mushrooms- A prominent source of nutriceuticals for the 21st century. Current Topics in Nutraceutical Research, 1, 257-280.

Chen, S. Y., Ho, K. J., Hsieh, Y. J., Wang, L. T., Mau, J. L. (2012). Contents of lovastatin, γ-aminobutyric acid and ergothioneine in mushroom fruiting bodies and mycelia. LWT- Food Science Technology, 47, 274-278. doi:10.1016/j.lwt.2012.01.019

Crozier, A., Jaganath I. B. & Clifford M. N. (2006). Phenols, polyphenols and Tannins: An overview. In A. Crozier, M. N. Clifford and H. Ashihara (Eds.), Plant secondary metabolites: Occurrence, structure and role in the human diet (pp. 2-3). Oxford, UK: Blackwell Publishing Ltd. doi:10.1002/9780470988558.ch1

Deiana, M., Rosa, A., Casu, V., Piga, R., Dessí, M. A., & Aruoma, O. I. (2004). L-ergothioneine modulates oxidative damage in the kidney and liver of rats in vivo: Studies upon the profile of polyunsaturated fatty acids. Clinical Nutrition, 23, 183-193. doi:10.1016/S0261-5614(03)00108-0

Ding, E. L, Hutfless, S. M., Ding, X., & Girotra, S. (2006). Chocolate and prevention of cardiovascular disease: A systematic review. Nutrition & Metabolism, 3. Retrieved from doi:10.1186/1743-7075-3-2 doi:10.1186/1743-7075-3-2

El-Mekkawy, S., Meselhy, M. R., Nakamura, N., Tezuka, Y., Hattori, M., Kakiuchi, N., Shimotohno, K., Kawahata, T., & Otake, T. (1998). Anti-HIV-1 and anti-HIV-1- protease substances from Ganoderma lucidum. Phytochemistry, 49, 1651-1657. doi:10.1016/S0031-9422(98)00254-4

Halpern, G. M. (2007). Mushroom cultivation. In G. M. Halpern (Ed.), Healing mushrooms: Effective treatments for today’s illnesses (pp. 121-123).

Garden City Park, New York, NY: Squares One Publishers.

Kohda, H., Tokumoto, W., Sakamoto, K., Fujii, M., Hirai, Y., Yamasaki, K., Komoda, Y., Nakamura, H., Ishihara, S., & Uchida, M. (1985). The biologically active constituents of Ganoderma lucidum (Fr.) Karst. Histamine release-inhibitory triterpenes. Chemical and Pharmaceutical Bulletin, 33, 1367-1374. doi:10.1248/cpb.33.1367

Komoda, Y., Shimizu, M., Sonoda, Y., & Sato, Y. (1989). Ganoderic acid and its derivatives as cholesterol synthesis inhibitors. Chemical and Pharmaceutical Bulletin, 37, 531-533. doi:10.1248/cpb.37.531

Lee, B. C., Bae, J. T., Pyo, H. B., Choe, T. B., Kim, S. W., Hwang, H. J., & Yun, J. W. (2004). Submerged culture conditions for the production of mycelia biomass and exopolysaccharides by the edible Basidiomycete Grifola frondosa. Enzyme and Microbiological Technology, 35, 369-376. doi:10.1016/j.enzmictec.2003.12.015

Lee I. K., & Yun B. S. (2006). Hispidin analogs from the mushroom Inonotus xeranticus and their free radical scavenging activity. Bioorganic and Medical Chemistry Letters, 16, 2376-2379. doi:10.1016/j.bmcl.2006.01.121

Li, S. P., Yang, F. Q., & Tsim, K. W. (2006). Quality control of Cordyceps sinensis, a valued traditional Chinese medicine. Journal of Pharmaceutical and Biomedical analysis, 41, 1571-1584. doi:10.1016/j.jpba.2006.01.046

Liu, X., Dong, M., Chen, X., Jiang, M., Lv, X., & Yan, G. (2007). Antioxidant activity and phenolics of an endophytic Xylaria sp. from Ginkgo biloba. Food Chemistry, 105, 548-554. doi:10.1016/j.foodchem.2007.04.008

Omar, N. A. M., Abdullah, N., Kuppusamy, U. R., Abdulla, M. A., & Sabaratnam, V. (2011). Nutritional composition, antioxidant capacity and antiulcer potential of Lentinus squarrosulus (Mont.) mycelia extract. Evidenced-Based Complementary and Alternative Medicine. doi:10.1155/2011/539356

Omar, N. A. M., Abdullah, S., Abdullah, N., Kuppusamy, U. R., Abdulla, M. A., & Sabaratnam, V. (2015). Lentinus squarrosulus (Mont.) mycelium enhanced antioxidant status in rat model. Drug Design, Development and Therapy, 9, 5957-5964.

Peng, I. W., & Kuo, S. M. (2003). Flavanoid structure affects the inhibition of lipid peroxidation in Caco-2 intestinal cells at physiological concentration. The Journal of Nutrition, 133, 2184-2187. doi:10.1093/jn/133.7.2184

Rahman, K. (2007). Studies on free radicals, antioxidants, and co-factors. Clinical Interventions in Aging, 2, 219-236.

Reiter, R. J. (1995). Oxidative process and antioxidative defense mechanism in aging brain. FASEB Journal, 9, 526-533. doi:10.1096/fasebj.9.7.7737461

Wanasundara, P. K. J. P. D., & Shahidi, F. (2005). Antioxidants: Science, technology, and applications. In F. Shahidi (Ed.), Bailey’s industrial oil and fat products (6th ed.), (pp. 431-489). New York, NY: John Wiley & Sons, Inc. doi:10.1002/047167849X.bio002

Wang, X. L., & Liu, Z. T. (2008). In vitro bacteriostasis of the intracellular ganoderic acids from the mycelium of Ganoderma lucidum. Food Science Technology, 10, 184-186.

Wu, G. S., Lu, J. J., Guo, J. J., Li, Y. B., Tan, W., Dang,Y. Y., Zhong, Z. F., Xu, Z. T., Chen, X. P., & Wang, Y. T. (2012) Ganodermic acid DM, a natural triterpenoid, induces DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells. Fitoterapia, 83, 408-414. doi:10.1016/j.fitote.2011.12.004

Wu, W., Yan, C., Li, L., Liu, Z., & Liu, S. (2004). Studies on the flavones using liquid chromatography-electrospray ionization tandem mass spectrometry. Journal of Chromatography A, 1047, 213-220. doi:10.1016/S0021-9673(04)01135-5

Xu, J. W., Zhao, W., & Zhong, J. J. (2010). Biotechnological production and application of ganoderic acids. Applied Microbiology and Biotechnology, 87, 457-466. doi:10.1007/s00253-010-2576-5

Yoshikawa, T., & Naito, Y. (2002). What is oxidative stress? Japan Medical Association Journal, 45, 271-276.

Yu, H. M., Wang, B. S., Huang, S. C., & Duh, P. D. (2006). Comparison of protective effects between Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. Journal of Agriculture and Food Chemistry, 54, 3132-3138. doi:10.1021/jf053111w

Zhu, M., Chang, Q., Wong, L. K., Chong, F. S., & Li, R. C. (1999) Triterpene antioxidants from Ganoderma lucidum. Phytotherapy Research, 13, 529-531. doi:10.1002/(SICI)1099-1573(199909)13:6<529::AID-PTR481>3.0.CO;2-X



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