ANTIBACTERIAL AND
ANTIOXIDANT ACTIVITIES OF FRACTIONS FROM GARCINIA
LATISSIMA MIQ. STEM BARK METHANOL EXTRACT
Neneng Siti Silfi AMBARWATI1,2, Berna ELYA2*, Amarila MALIK2, Muhammad HANAFI3,4, Khairinisa LESTARI2, Nuraini PUSPITASARI2, Apriantika SARI2, Rut
Juliany TARIGAN2, Hanita OMAR5
1 Faculty of Engineering, Universitas Negeri Jakarta, Jl. Rawamangun Muka, East Jakarta 13220, Indonesia.
2 Faculty of Pharmacy, Universitas Indonesia, UI Depok Campus, Depok 16424, Indonesia.
3 Research Center for Chemistry, Indonesian Institute of Sciences (LIPI), Kawasan PUSPIPTEK, Serpong, Tangerang, 15314, Indonesia.
4 Faculty of Pharmacy, University of Pancasila, Srengseng Sawah, Jakarta, Indonesia.
5 Chemistry Division, Centre for Foundation Studies in Science, University of Malaya
* Corresponding author. E-mail: [email protected]
Abstract
Objective. This study aimed to get complete information about the antioxidant and antibacterial against B. subtilis and Staphylococcus aureus activities of the fractions from G. latissima Miq. stem bark methanol extract.
Methods. Fractionation was performed by column chromatography. The antibacterial activities of the fractions from Garcinia latissima Miq. stem bark were assayed by
inhibition zone technique, bioautography, and minimum inhibition concentration. The antioxidant activity were evaluated using DPPH (2,2-diphenyl-1picrylhydrazyl) and
FRAP (Ferric Reducing Antioxidant Power) methods. Results. Stem bark methanol extract had higher antioxidant activity (% inhibition = 95.68%) than ethyl acetate
extract and n-hexane stem bark extract of G. latissima Miq. The stem bark methanol extract obtained ten fractions. The greatest of inhibition zone diameter against B.
subtilis (ATCC (the American Type Culture Collection) 6633) and S. aureus (ATCC 25923) was fraction G (7.83 ± 0.46 mm and 7.43 ± 0.15 mm). The highest antioxidant activity fraction by DPPH method and FRAP method was fraction G. Phytochemical screening showed that fraction G contained flavonoid and tannin. Conclusion. The results obtained reveal that the fraction G of stem bark of G. latissima Miq. methanol extract possessed antibacterial activity against B. subtilis and S. aureus and
antioxidant activity.
Keywords: Garcinia latissima Miq. stem bark. methanol fractions. Bacillus subtilis.
Staphylococcus aureus. DPPH & FRAP.
Previous research had been conducted on G. latissima Miq. stem bark methanol extract that the extract (20,000 ppm) was active against B. subtilis bacterium with inhibition zone diameter was 10.70±0.638 mm, and the inhibition zone diameter against S. aureus was 10.38±0.653 mm [1]. The MIC (Minimum Inhibitory
Concentration) and MBC (Minimum Bactericidal Concentration) values of the extract against B. subtilis were 625 and 5,000 ppm [1]. The MIC and MBC values of the extract against S. aureus were 2,500 and 5,000 ppm [1]. This study was continued with
fractionation and tested the fractions against B. subtilis and S. aureus bacteria.
Antioxidant testing was also carried out on G. latissima Miq extracts and fractions.
Antioxidant Activity Test for Extract
Antioxidant Activity Test of Fractions with DPPH Method
The antioxidant activity test of fraction G obtained that IC50 value of 5.10 μg/mL.
Antioxidant Activity Test for Extract with DPPH method.
Fractionation of The Most Active from Garcinia latissima Miq. Stem Bark Extract by using column chromatography.
Inhibition Zone Test for Fractions, each fraction was dropped on a sterile 6 mm
diameter paper disk placed on a petri dish containing agar media containing bacteria.
Incubate at 37 ° C for 24 hours and measure the diameter of the inhibition zone using the calipers [7].
Antioxidant Activity Test Fractions with DPPH method [8][9]
% inhibition = Absorbance of blank – Absorbance of sample x 100%
Absorbance of blank
Antioxidant Activity Test with FRAP method [10]:
FeEAC (�mol/gram) = _____ΔA___ x Av x D x ____1__ x 105 GRAD (AFS) Spv Csample
Phytochemical Screening of The Most Active Fraction as Antioxidant [11]
The test was performed for the identification of flavonoids (AlCl3), saponins (hot water and HCl), tannins (lead acetate) [12].
Introduction
Conclusion
Results and Discussions
Materials and Methods
Antioxidant Activity Test of Fractions with FRAP Method
Phytochemical Screening of The Most Active Fraction as Antioxidant
Fraction G showed a positive flavonoid test [5]. The hydroxyl groups found in A and / or B rings of flavonoids can increase antioxidant capacity, especially hydroxyl groups at ortho positions at C3 'and C4' [6]. Fraction G shows positive results of tannin
compounds.
The results obtained reveal that the fraction G of stem bark of G. latissima Miq.
methanol extract possessed the highest antibacterial activity against B. subtilis and S.
aureus and highest antioxidant activity. The fraction G contained flavonoids and tannins.
The research supported by “Hibah Tugas Akhir Doktor (TADOK) Universitas Indonesia 2018”.
Acknowledgments
References
[1] N. S. S. Ambarwati, B. Elya, A. Malik, and M. Hanafi, “Evaluation of Antimicrobial Activities of Garcinia latissima Miq. Stem Bark Extract,” J. Young Pharm., vol. 9, no. 1, pp. 118–121, 2017.
[2] P. A. Murray, R. K., Bender, D. A., Botham, K. M., Kennelly, P. J., Rodwell, V. W., & Weil, Biologic Oxidation. Harper’s Illustrated Biochemistry, 28th Edition. 2009.
[3] A. Sarma, A. Mallick, and A. Ghosh, “Free Radicals and Their Role in Different Clinical Conditions : An Overview,” Int. J. Pharma Sci. Res., vol. 1, no. 3, pp. 185–192, 2010.
[4] R. Sayuti, K., & Yenrina, Antioksidan Alami dan Sintetik. (T. Anggraini, Ed.) (1st ed.). Padang: Andalas University Press., 2015.
[5] E. Iqbal, K. Abu, and L. B. L. Lim, “Phytochemical screening , total phenolics and antioxidant activities of bark and leaf extracts of Goniothalamus velutinus ( Airy Shaw ) from Brunei Darussalam,” J. King Saud Univ. - Sci., vol. 27, no. 3, pp. 224–232, 2015.
[6] I. Fidrianny, L. Filkayuniar, and M. Insanu, “Antioxidant activities of various seed extracts from four varieties of rambutan (Nephelium lappaceum) using 2,2-diphenyl-1-picrylhydrazyl and 2,2’-azinobis (3- ethylbenzothiazoline-6-sulfonic acid) assays,” Asian J. Pharm. Clin. Res., vol. 8, no. 5, pp.
227–231, 2015.
[7] K. P. Kochuthressia, S. J. Britto, Jaseentha, and R. Raphael, “In Vitro Antimicrobial Evaluation of Kaempferia galanga L. Rhizome Extract,” Am. J.
Biotechnol. Mol. Sci., vol. 2, no. 1, pp. 1–5, 2012.
[8] P. Molyneux, “The use of the stable free radical diphenylpicryl- hydrazyl (DPPH) for estimating antioxidant activity,” Songklanakarin J. Sci.
Technol., vol. 26, no. December 2003, pp. 211–219, 2004.
[9] V. Seidel, “Initial and Bulk Extraction,” in Natural Product Isolation, 2nd ed., S. D. Sarker, Z. Latif, and A. I. Gray, Eds. New Jersey: Humana Press, 2006, pp. 27–46.
[10] C. Wong, W. Cheung, Y. Lau, A. Bolanos de la Torre, and R. Owusu-Apenten, “A FRAP Assay at pH 7 unveils Extra Antioxidant Activity from Green, Black, White and Rooibos Tea but not Apple Tea,” Food Nutr. Rep., vol. 1, no. 1, pp. 1–8, 2015.
[11] H. Wagner and S. Bladt, Plant Drug Analysis: A Thin Layer Chromatography Atlas. Analytica Chimica Acta, 2001.
[12] P. O. Ukoha, E. A. C. Cemaluk, O. L. Nnamdi, and E. P. Madus, “Tannins and other phytochemical of the Samanaea saman pods and their antimicrobial activities,” African J. Pure Appl. Chem., vol. 5, no. August, pp. 237–244, 2011.
