UJI PENANGKAPAN RADIKAL 2,2-difenil-1-pikrilhidrazil (DPPH) OLEH EKSTRAK ETANOL BUNGA KECOMBRANG (Nicolaia speciosa (Bl.) Horan) DAN BUAH TALOK (M. calabura L. )

  • Tatang Irianti Gadjah Mada University
  • Hari Purnomo Faculty of Vacation School, Gadjah Mada University, Yogyakarta
  • Kuswandi Kuswandi Faculty of Vacation School, Gadjah Mada University, Yogyakarta
  • Sindu Nuranto Faculty of Vacation School, Gadjah Mada University, Yogyakarta
  • Damiana Nitya Kanistri Faculty of Pharmacy, Gadjah Mada University, Yogyakarta
  • Yosi Bayu Murti Faculty of Pharmacy, Gadjah Mada University, Yogyakarta
  • Sofa Farida Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional
Keywords: kecombrang (Nicolaia speciosa (Bl.) Horan), talok (M. calabura L.), antioxidant, DPPH

Abstract

ABSTRACT

The process of excessive free radicals  in our body plays a major part in the development of chronic and degenerative illness such as cancer, autoimmune disorders, aging, cataract, rheumatoid arthritis, cardiovascular and neurodegenerative diseases. The recently study reported that free radical and oxidants play a dual role as both toxic and beneficial compounds, since they can be either harmful or helpful to the body. Therefore, natural antioxidants can be explored to prevent degenerative diseases and in the present paper we have investigated antioxidant activity of extracts from Nicolaia speciose (Bl.) Horan flower and Muntingia calabura L.  fruit for its free radical scavenging activity using 2,2-diphenyl, 1-picryl hydrazil (DPPH) radical scavenging activity. The results revealed that both ethyl acetate fractions of N.  speciaose and M. calabura have the highest antioxidant activitiy with IC50 29.81 and 14.48 µg/ml respectively.   The antioxidant activity of both ethanolic extracts were more potent than hexan and water fractions with IC50 39.27 and 137.20 µg/ml respectively. The investigation of IC50 values indicated that the antioxidant activity show moderate to very active. Active compounds were identified using thin layer chromatography with FeCl3, AlCl3, 2,4-DNPH and anisaldehide-H2SO4 spray reagents. Chromatogram of ethylacetate fraction showed that spots on the hRf 12 and 56 were thought to contain phenolic compounds with a carbonyl group, while the hRf 37 was suspected flavone compounds with 3-OH group and the hRf 50 was alleged the ortho-dihydroxy flavone or ortho hydroxy and free carbonyl. Chromatogram of ethyl acetate of talok fruits fraction showed that spots on the hRf 19.31 and 44 were suspected flavone compounds with ortho-hydroxy-carbonyl group and or ortho-hydroxy.

ABSTRAK

Radikal bebas berlebih dalam tubuh dapat memicu tumbuhnya sel kanker, penyumbatan pembuluh jantung, kerusakan oksidatif otak dan penuaan dini. Hal ini menyebabkan penelitian tentang potensi antioksidan terus mengalami peningkatan, terutama antioksidan alami dari tanaman. Bunga kecombrang (Nicolaia speciosa (BI.) Horan) dan buah talok (Muntingia Calabura L.) dilaporkan aktif sebagai antioksidan dan mengandung senyawa aktif dengan tingkat kepolaran yang berbeda. Oleh karena itu, penelitian ini bertujuan untuk menguji aktivitas dengan penangkapan radikal bebas fraksi-fraksi ekstrak bunga kecombrang dan buah talok serta karakterisasi golongan senyawa aktifnya. Hasil fraksinasi ekstrak diuji dengan larutan DPPH 0,4 mM dan diukur dengan spektrofotometer UV-Vis pada panjang gelombang 517 nm. Hasil absorbansi sampel dan kontrol diolah untuk mendapatkan persen penangkapan radikal DPPH dan aktivitas antioksidan dievaluasi melalui nilai IC50. Senyawa aktif diidentifikasi menggunakan kromatografi lapis tipis dengan penampak bercak FeCl3, AlCl3, 2,4-DNPH dan anisaldehid-H2SO4.  Fraksi-fraksi ekstrak bunga kecombrang dan buah talok menunjukkan tingkat kekuatan antioksidan antara sedang sampai sangat aktif. Fraksi etil asetat bunga kecombrang serta buah talok memiliki aktivitas penangkapan radikal bebas tertinggi dengan IC50 sebesar 29,81 µg/ml dan 14,48 µg/ml. Pada fraksi etil asetat ini, buah talok mempunyai potensi aktivitas antioksidan dua kali lipatnya dari bunga kecombrang.  Fraksi air, ekstrak etanol serta fraksi heksan bunga kecombrang memiliki IC50 sebesar 39,27 µg/ml, 44,08 µg/ml, dan 135,36 µg/ml. Sedangkan ekstrak etanol, fraksi air serta fraksi heksan buah talok memiliki IC50 sebesar 137,20 µg/ml, 282,47 µg/ml, dan 2611,70 µg/ml. Hasil KLT fraksi etil asetat buah kecombrang menunjukkan bahwa bercak pada hRf 12 dan 56 mengandung senyawa fenolik dengan gugus karbonil, sedangkan hRf 37 diduga senyawa flavon dengan gugus 3-OH dan pada hRf 50 merupakan flavon dengan o-dihidroksi dan atau o-hidroksi karbonil bebas. Kemudian hasil KLT fraksi etil asetat buah talok menunjukkan bahwa bercak dengan nilai hRf  19, 31, dan 44 merupakan flavon dengan gugus o-hidroksi karbonil dan atau gugus o-dihidroksi. Bercak pada hRf 31 juga mengindikasikan adanya senyawa terpenoid.

 

 

References

DAFTAR PUSTAKA

Bors, W., Hellen, W., Michel, C., & Saran, M. (1990). Flavonoids as Antioxidants: Determination on Radical-Scavenging Effeciencies. Methods Enzymol. Vol. 186, pp. 343-355. https://doi.org/https://doi.org/10.1016/0076-6879(90)86128-I

Foti, M., Piattelli, M., Baratta, M. T., Ruberto, G., Chimiche, S., Doria, V. A., & Catania, I. (1996). Flavonoids, Coumarins, and Cinnamic Acids as Antioxidants in a Micellar System. Structure-Activity Relationship, J.Agric. Food. Chem., 44(2), 497-500. https://doi.org/10.1021/jf950378u

Gordon, I. (1994). Functional Food, Food Design, Pharmafood. New York: Champman and Hall.

Haleagrahara N., Jackie T., Chakravarthi S., R. M. and K. A. (2010). Protective Effect of Etlingera elatior (torch ginger) Extract On Lead Acetate – Induced Hepatotoxicity In Rats. The Journal of Toxicological Sciences, 35(5): 663-671.

Harborne, J. (1987). Metode Fitokimia (Cetakan IV). Bandung: ITB.

Jackie, T., Haleagrahara, N., & Chakravarthi, S. (2011). Antioxidant Effects of Etlingera elatior Flower Extract Against Lead Acetate-Induced Pertubations in Free Radical Scavenging Enzymes and Lipid Peroxidation in Rats. BMC Research Notes 4, 67. https://doi.org/10.1186/1756-0500-4-67

Lachumy, S.T.J., S. Sasidharan, V. Sumathy, & Z. Zuraini. (2010). Pharmacological activity, phytochemical analysis and toxicity of methanol extract of Etlingera elatior (torch ginger) flowers. Asian Pacific Journal of Tropical Medicine 3: 769-774. https://doi.org/10.1016/S1995-7645(10)60185-X

Jork, H., Funk, W., Fischer, W., Wimmer, H. (1990). Thin-Layer Chromatography: Reagents and Detection Methods (Volume 1a). Germany: VCH Publishing.

Mikova K. (2001). The Regulations of Antioxidant in Food. CRC Press. Wood-head Publishing.

Preethi. K, et al. (2010). In Vitro Activity of Extracts from Fruits of M. calabura Linn. From India, Pharmacognosy Journal, 2, 11-18.

Mabry, T. J., Markham, K. R. and Thomas, M. B. (1970). The Systematic Identification of Flavanoids, Springer Verlag: Berlin

Markham, K. R. (1988). Cara Mengidentifikasi Flavonoid. Bandung: ITB.

Maslarova, N. V. Y. (2001). Inhibiting Oxidation. Cambridge: Woodhead Publishing Limited.

Molyneux, P. (2004). The Use of Stable Free Radical Diphenylpicrylhydrazyl (DPPH) for Estimating Antioxidant Activity, J. Sci. Technicol., 26(2), 211-219.

Ozyurt, D., Demirata, B., & Apak, R. (2007). Determination of total antioxidant capacity by a new spectrophotometric method based on Ce (IV) reducing capacity measurement (1)Diakses tanggal 18 Mei 2010.

https://doi.org/10.1016/j.talanta.2006.06.015

Masoko, P., dan Eloff, J.N. (2007). Screening of Twenty-four South African Combertum and Six Terminalia Species (Combretaceae) for Antioksidant Activities, African Journal of Traditional, Complimentary and Alternative Medicines, 4 (2), 231-239.

Pokorny, J., Yanishlieva, N. and Gordon, M. (2001). Antioxidant in Food: Practical Applications. New York: CRC Press.

Robinson, T. (1995). Kandungan Organik Tumbuhan Tinggi. Bandung: ITB.

Kumar S. (2010). Extraction of Essential Oil Using Steam Distillation. Rourkela: National Institute of Technology Press.

Siddiqua, et al. (2010). Antioxidant Activity and Estimation of Total Phenolic Content of M. calabura L. by Colorimetry, International Journal of ChemTech Research, 2 (1), 205-208.

Su, Bao Ning, et al. (2003). Activity Guided Isolation of The Chemical Constituents of M. calabura. L Using a Quinone Reductase Induction Assay. Phytochemistry, 63, 335–341.

Wagner, H., Bladt, S. (1996). Plant Drug Analysis. New York: Springer-Verlag Berlin Heidelberg.

Zakaria, Z. A, el al. (2010). In vitro Antimicrobial Activity of M. calabura L. Extracts and fractions. Internasional Journal of Pharmacology, 4, 304-308.

Published
2019-08-30