Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

In Vitro Antioxidant Activity of Alnus firma Extracts

사방오리(Alnus firma) 추출물의 in vitro 항산화 활성

  • Choi, Hye Jung (Department of Biology and Chemistry, Changwon National University) ;
  • Joo, Woo Hong (Department of Biology and Chemistry, Changwon National University)
  • 최혜정 (창원대학교 생물학화학융합학부) ;
  • 주우홍 (창원대학교 생물학화학융합학부)
  • Received : 2018.12.12
  • Accepted : 2019.01.14
  • Published : 2019.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

This study evaluated the antioxidant activity of the extract and fractions of Alnus firma. Alnus firma had the highest total phenolic content ($452.80{\pm}7.01{\mu}g$ gallic acid equivalents/mg) in a methanol (MeOH) fraction and the highest total flavonoid content ($112.29{\pm}11.14{\mu}g$ rutin equivalents/mg) and antioxidant capacity ($936.23{\pm}0.07{\mu}g$ ${\alpha}$-tocopherol equivalents/mg) in an ethylacetate (EA) fraction. The antioxidant activities of various solvent extract fractions of Alnus firma were evaluated using various antioxidant assays, including ${\beta}$-carotene-linoleate assay, reducing power assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, metal chelating activity assay, superoxide anion radical scavenging assay, and lipid peroxidation inhibition assay using the ferric thiocyanate method. These activities were compared with those of ascorbic acid, butylated hydroxyanisole (BHA), gallic acid (GA), butylated hydroxytoluene (BHT), and ${\alpha}$-tocopherol. First, at a $250{\mu}g/ml$ concentration, the EA and MeOH fractions of A. firma showed 92.43% and 89.20% DPPH radical scavenging activity, respectively. Second, $50{\mu}g/ml$ of the EA fraction exhibited 72.49% superoxide anion radical scavenging activity, a little greater than the same dose of GA (60.88%). Finally, 0.5 and 1 mg/ml of the EA fraction showed 73.45% and 73.29% inhibition of peroxidation in the ${\beta}$-carotene-linoleic acid system, respectively. The decreasing order of reducing power was EA fraction > n-butanol (BuOH) fraction > dichloromethane (DCM) fraction > n-hexane (HX) fraction. The results obtained in the present study indicated that Alnus firma can be used as an easily accessible potential source of natural antioxidants.

본 연구는 사방오리 추출 분획물의 항산화 활성을 검증하기 위하여 시행되었다. 사방오리는 메탄올(MeOH) 분획에 가장 높은 총 페놀화합물($452.80{\pm}7.01{\mu}g$ gallic acid equivalents/mg)을 그리고 에틸에세테이트(EA) 분획 에 가장 높은 총 플라보노이드 함량($112.29{\pm}11.14{\mu}g$ rutin equivalents/mg)과 가장 높은 총 항산화 활성($936.23{\pm}0.07{\mu}g$ ${\alpha}$-tocopherol equivalents/mg)을 가지고 있었다. 사방오리의 용매 추출분획물에 대한 항산화 활성은 아스코빅산 등 표준 물질과 비교하여 1,1-diphenyl-2-picrylhydrazyl (DPPH) 라디칼 소거능 측정, 슈퍼옥사이드 음이온 라디칼 소거능 측정, ${\beta}$-carotene-linoleate 에세이, 환원력 측정, ferric thiocyanate법으로 지질과산화 억제력 측정 그리고 금속 킬레이팅능 측정 등 다양한 항산화 활성을 측정하였다. $250{\mu}g/ml$ 농도의 사방오리 EA와 MeOH 분획은 DPPH 라디칼 소거능 측정에서 각각 92.43와 89.20% 항산화 활성을 가지고 있었다. $50{\mu}g/ml$의 EA분획은 72.49% 슈퍼옥사이드 음이온 라디칼 소거능을 보였으며 이는 gallic acid의 소거능(60.88%)보다 조금 높았다. 또한 ${\beta}$-carotene-linoleic acid 시스템에 의한 과산화 억제능 조사에서 0.5과 1 mg/ml 농도의 EA 분획이 각각 73.45와 73.29% 억제능을 보였다. 항산화 활성은 EA 분획 > BuOH 분획 > DCM분획 > HX분획 순 이였다. 이상의 본 연구 결과에서 사방오리는 천연 항산화제제 개발을 위한 용이하게 접근 가능한 잠재적인 자원으로 판단된다.

Keywords

SMGHBM_2019_v29n2_231_f0001.png 이미지

Fig. 1. Free radical scavenging activity of MeOH, HX, DCM and EA fractions of Alnus firma by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method.

SMGHBM_2019_v29n2_231_f0002.png 이미지

Fig. 2. Superoxide anion radical scavenging activity of MeOH, HX, DCM and EA fractions of Alnus firma by PMS/ NADH-NBT method.

SMGHBM_2019_v29n2_231_f0003.png 이미지

Fig. 3. Antioxidant activities of Alnus firma extracts using the β-carotene linoleate model system. L-ascorbic acid and BHA were used as positive controls.

SMGHBM_2019_v29n2_231_f0004.png 이미지

Fig. 4. Reducing power of MeOH, HX, DCM, EA and BuOH fractions of Alnus firma.

SMGHBM_2019_v29n2_231_f0005.png 이미지

Fig. 5. Inhibition of lipid peroxidation of Alnus firma extracts. Ferric thiocyanate (FTC) method was used for determination of lipid peroxidation.

SMGHBM_2019_v29n2_231_f0006.png 이미지

Fig. 6. Metal chelating effect of organic solvent-fractions of Alnus firma.

Table 1. Extraction yields, total phenolic contents, total flavonoid contents, total antioxidant contents and total reducing power in crude Alnus firma extracts

SMGHBM_2019_v29n2_231_t0001.png 이미지

References

  1. Bae, C. I., Gong, J. M., Oh, J. W., Kim H. J., Oh, G. J., Park, S. K., Chung, S. G. and Cho, E. H. 1997. Studies on the cytotoxic constituent of Alnus hirsuta. J. Pharmaceut. Soc. Korea 41, 559-564.
  2. Branen, A. S. 1975. Toxicology and biochemistry of butylated hydroxyanisole and butylated dihydroxytoluene. J. Am. Oil Chem. Soc. 52, 59-63. https://doi.org/10.1007/BF02901825
  3. Choi, H. J., Jeong, Y. K., Kang, D. O. and Joo, W. H. 2008. Inhibitory effects of four solvent fractions of Alnus firma on ${\alpha}$-amylase and ${\alpha}$-glucosidase. J. Life Sci. 18, 1005-1010. https://doi.org/10.5352/JLS.2008.18.7.1005
  4. Decker, E. A. and Welch, B. 1990. Role of ferritin as a lipid oxidation catalyst in muscle food. J. Agr. Food Chem. 38, 674-677. https://doi.org/10.1021/jf00093a019
  5. Hatano, T., Edamatsu, R., Hiramatsu, M., Mori, A., Fujita, Y., Yasuhara, T., Yoshida, T. and Okuda, T. 1989. Effect of the interaction of tannins with co-existing substances VI. Effects of tannins and related polyphenol on superoxide anion radical and on 1,1-diphenyl-2-picrylhydrazyl radical. Chem. Pharm. Bull. 37, 2016-2021. https://doi.org/10.1248/cpb.37.2016
  6. Hsu, B., Coupar, I. M. and Ng, K. 2006. Antioxidant activity of hot water extract from the fruit of the Doum palm, Hyphaene thebaica. Food Chem. 98, 317-328. https://doi.org/10.1016/j.foodchem.2005.05.077
  7. Hyun, S. H., Jung, S. K., Jwa, M. K., Song, C. K., Kim, J. H. and Lim, S. B. 2007. Screening of antioxidants and cosmeceuticals from natural plant resources in Jeju island. Kor. J. Food Sci. Technol. 39, 200-208.
  8. Karchesy, J. J., Laver, M. L., Barofsky, D. F. and Barofsky, E. 1974. Structure of oregonin, a natural diarylheptanoid xyloside. J. Chem. Soc. Chem. Commun. 16, 649-650.
  9. Kim, M. B., Park, J. S. and Lim, S. B. 2010. Antioxidant activity and cell toxicity of pressurised liquid extracts from 20 selected plant species in Jeju, Korea. Food Chem. 122, 546-552. https://doi.org/10.1016/j.foodchem.2010.03.007
  10. Lee, C. B. 1980. Plant flora of Korea. Hyangmoonsa.
  11. Lee, Y. A., Kim, K. H., Kim, J. S., Cho, S. M., Kim, S. W. and Lee, M. W. 2000. Antioxidative effects of diarylheptanoids from Alnus firsuta. J. Pharmaceut. Soc. Korea 44, 193-196.
  12. Maeura, Y., Weisburger, J. H. and Williams, G. 1984. Dosedependent reduction of N-2-fluorenylacetamide-induced liver cancer and enhancement of bladder cancer in rats by butylated hydroxytoluene. Cancer Res. 44, 1604-1610.
  13. Miller, H. E. 1971. A simplified method for the evaluation of antioxidants. J. Am. Oil Chem. Soc. 48, 91-91. https://doi.org/10.1007/BF02635693
  14. Nishikimi, M., Rao, N. A. and Yagi, K. 1972. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem. Bioph. Res. Co. 46, 849-854. https://doi.org/10.1016/S0006-291X(72)80218-3
  15. Oktay, M., Gulcin, I. and Kufrevioglu, O. I. 2003. Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts. LWT-Food Sci. Technol. 36, 263-271. https://doi.org/10.1016/S0023-6438(02)00226-8
  16. Osawa. T. and Namiki, M. 1981. A Novel type of antioxidant isolated from leaf wax of Eucalyptus leaves. Agr. Biol. Chem. 45, 735-739. https://doi.org/10.1271/bbb1961.45.735
  17. Oyaizu, M. 1986. Studies on products of browning reaction-Antioxidative activities of products of browning reaction prepared from glucoamine. Jpn. J. Nutr. Diet. 44, 307-315. https://doi.org/10.5264/eiyogakuzashi.44.307
  18. Prieto, P., Pineda, M. and Aguilar, M. 1999. Spectophotometric quantitaion of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal. Biochem. 269, 337-341. https://doi.org/10.1006/abio.1999.4019
  19. Sajid, M., Khan, M. R., Shah, N. A., Shah, S. A., Ismail, H., Younis, T. and Zahra, Z. 2016. Phytochemical, antioxidant and hepatoprotective effects of Alnus nitida bark in carbon tetrachloride challenged Sprague Dawley rats. BMC Complem. Altern. Med. 16, 268. https://doi.org/10.1186/s12906-016-1245-3
  20. Sakanaka, S., Tachibana, Y. and Okada, Y. 2005. Preparation and antioxidant properties of extracts of Jaoanese persimmon leaf tea (kakinoha-cha). Food Chem. 89, 569-575. https://doi.org/10.1016/j.foodchem.2004.03.013
  21. Sheth, K., Bianchi, E., Wiedhopf, R. and Cole, J. R. 1973. Antitumor agents from Alnus oregona. (Betulaceae). J. Pharmaceut. Sci. 62, 139-140. https://doi.org/10.1002/jps.2600620129
  22. Slinkard, K. and Singleton, V. L. 1997. Total phenol analyses: Automation and comparison with manual methods. Am. J. Enol. Viticult. 28, 49-55.
  23. Suga, T., Iwata, N. and Asakawa, Y. 1972. Chemical constituents of male flower of Alnus pendula (Betulaceae). Bull. Chem. Soc. Jpn. 45, 2058-2060. https://doi.org/10.1246/bcsj.45.2058
  24. Woisky, R. G. and Salatino, A. 1998. Analysis of propolis: some parameters and procedures for chemical quality control. J. Apicult. Res. 37, 99-105. https://doi.org/10.1080/00218839.1998.11100961