Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effect of Graviola Leaves Extracts on Antioxidant Activity and Melanin Production

멜라닌 생성 및 항산화 활성에 미치는 그라비올라 잎 추출물의 영향

  • Lee, Kyuwon (Department of Chemistry, Dong-Eui University) ;
  • Jang, Jiwon (Department of Chemistry, Dong-Eui University) ;
  • Park, Sumin (Department of Chemistry, Dong-Eui University) ;
  • Kang, Kihun (Department of Chemistry, Dong-Eui University) ;
  • Yoon, Hyesoo (Department of Chemistry, Dong-Eui University) ;
  • Ha, Yejin (Department of Chemistry, Dong-Eui University) ;
  • Jeon, Sojeong (Department of Chemistry & Biology, Dong-Eui University) ;
  • Ko, Hye Ju (Department of Chemistry & Biology, Dong-Eui University) ;
  • Kim, Moon-Moo (Department of Applied Chemistry, Dong-Eui University) ;
  • Oh, Yunghee (Department of Applied Chemistry, Dong-Eui University)
  • Received : 2017.11.16
  • Accepted : 2018.01.22
  • Published : 2019.06.30
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Abstract

The purpose of this study was to investigate the effect of methanolic extracts of graviola, Annona muricate leaves (AMME) on antioxidant activity and melanin production. First of all, DPPH radical and reducing power were performed to determine the antioxidant effect of AMME and organic solvent fractions. AMME and organic solvent fractions showed antioxidative activity in a concentration dependent manner. The ethyl acetate fraction of AMME among organic solvent fractions showed the highest antioxidant activity. Moreover, tyrosinase activity was performed to confirm the effect of organic fractions on melanin production. AMME, ethyl acetate, and hexane fractions increased tyrosinase activity a dose dependent manner. Next, the hexane fraction with the best effect on melanin synthesis in AMME organic solvent fraction was divided into 12 fractions by silica column chromatography. Among them, the fraction 7 and 8 showed the highest DPPH radical scavenging activity and reducing power. In addition, the fraction 7 and 8 at $64{\mu}g/ml$ showed melanin synthesis by 260% and 184%, respectively. Finally, the fraction 8 at $4{\mu}g/ml$ showed melanin synthesis by 34% in B16F1 cells. LC-MS analysis showed that fraction 7 and fraction 8 have a molecular weight of 617 and 619, respectively. FT-IR analysis showed that fractions 7 and 8 is similar to bis(2-hydroxyethly)dimerate. Above results suggest that graviola leaves extracts could be applicable to the development of natural antioxidants or hair cosmetics which are related to the promoting effect of melanin production.

본 연구의 목적은 그라비올라 잎의 항산화 효능과 멜라닌 생성 촉진효과를 조사하는 것이다. 먼저 AMME와 유기용매 별 분획물의 항산화 효능을 확인하기 위해 DPPH radical 소거능과, reducing power를 수행하였다. AMME와 유기용매별 분획물은 농도에 비례하여 항산화 효능이 증가하였으며, 그 중 ethyl acetate 분획물에서 가장 높은 항산화 효능을 보였다. 항산화 효능을 보인 분획물들의 멜라닌 생성에 미치는 영향을 확인하기 위해 tyrosinase activity를 수행한 결과 AMME, ethyl acetate, hexane 분획물에서 농도에 비례하여 멜라닌 생성 촉진효능이 나타났다. 그 중에서 AMME 유기 용매 별 분획물에서 멜라닌 합성 효능이 가장 좋은 hexane 분획물을 칼럼크로마토그래피를 통해 12가지 분획물로 나누어 항산화 실험과 멜라닌 합성실험을 수행하였다. 그 중 Fig. 8에서 DPPH radical 소거능과 환원력이 가장 우수한 것으로 나타났다. In vitro melanin 합성 실험 결과, Fr. 7은 $64{\mu}g/ml$ 농도에서 260%의 멜라닌 합성 효능을 보였고, Fr. 8은 $64{\mu}g/ml$ 농도에서 184%의 멜라닌 합성 효능이 관찰되었다. 최종적으로 B16F1 세포에서 Fr. 8의 $4{\mu}g/ml$ 농도에서 34%의 멜라닌 합성 효능을 보였다. LC-MS 결과 Fr. 7이 617의 분자량을 나타냈고, Fr. 8이 분자량 619의 분자량을 나타냈다. FT-IR 결과 Fig. 7과 Fig. 8 모두 Bis(2-hydroxyethly)dimerate와 비슷한 유효성분을 나타냈다. 이상의 결과로 그라비올라 잎은 천연 항산화제 또는 멜라닌 생성 촉진효과와 관련 있는 모발 제품 개발에 응용가능성이 있다고 판단된다.

Keywords

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Fig. 1. Schematic diagram of extraction procedure, antioxidant activity and melanin synthesis test of Annona muricata.

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Fig. 2. Standard curve and total polyphenol content of Fr. 7 and Fr. 8 in silica chromatography with hexane fraction.

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Fig. 3. LC-MS spectrum of Fr. 7 and Fr. 8. LC-MS spectrum was analyzed by the Agilent 6,100 Series LC/MS. Peaks were confirmed that the molecular weights are 617 and 619.

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Fig. 4. IR spectrums of Fr. 7 and Fr. 8. IR spectrum was analyzed by the NICOLET iS10.

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Fig. 5. The effect of AMME and treatment groups (E.A. Fr., Hexane. Fr., Fr.7 and Fr.8) on scavenging activity of DPPH radical.

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Fig. 6. The effect of AMME and treatment groups (E.A. Fr., Hexane. Fr., Fr. 7, Fr. 8) on reducing power.

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Fig. 7. Effects of AMME and treatment groups (E.A. Fr. and Hexane. Fr.) on tyrosinase activity.

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Fig. 8. Hexane fraction of AMME on cytotoxicity.

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Fig. 9. Effect of hexane fraction of AMME on melanin synthesis by DOPA-oxidation using tyrosinase.

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Fig. 10. Effect of Fr. 8 in silica chromatography with hexane fraction on melanin synthesis in B16F1 Cells.

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