PENGUJIAN TOKSISITAS IN VITRO EKSTRAK DAN FRAKSI DARI DAUN JAMBU AIR (SYZYGIUM AQUEUM) DAN KULIT BUAH DELIMA (PUNICA GRANATUM) TERHADAP SEL VERO
DOI:
https://doi.org/10.5614/api.v42i2.5527Abstract
Daun jambu air (Syzygium aqueum) dan kulit buah delima (Punica granatum) merupakan tanaman yang pada umumnya terdapat di Indonesia. Kandungan flavonoid pada daun jambu air dan kulit buah delima diketahui memiliki efek antioksidan yang baik untuk tubuh sehingga tanaman ini banyak digunakan untuk pengobatan dan pencegahan beberapa penyakit. Secara tradisional daun jambu air dan kulit buah delima biasa digunakan sebagai antimikroba, antidiabetes, dan pengobatan diare. Penelitian ini bertujuan untuk menentukan toksisitas ekstrak dan fraksi dari daun jambu air dan kulit buah delima secara in vitro terhadap sel Vero. Serbuk simplisia daun jambu air diekstraksi bertingkat dengan metode maserasi, sedangkan serbuk simplisia kulit buah delima diekstraksi bertingkat menggunakan metode refluks. Keduanya diekstraksi bertingkat dengan pelarut n-heksana, etil asetat, dan etanol. Ekstrak dipantau menggunakan kromatografi lapis tipis (KLT). Ekstrak etanol dari daun jambu air dan kulit buah delima difraksinasi dengan metode ekstraksi cair-cair (ECC) menggunakan pelarut n-heksan dan etil asetat. Fraksi dipantau menggunakan kromatografi lapis tipis (KLT). Ekstrak dan fraksi diuji toksisitasnya secara in vitro terhadap sel Vero menggunakan reagen alamar blue. Ekstrak dan fraksi daun jambu air dan kulit buah delima pada konsentrasi uji antara 2 g/ml sampai dengan 1024 g/ml bersifat tidak toksik terhadap sel Vero. Berdasarkan penelitian, dapat disimpulkan bahwa ekstrak n-heksana, etil asetat, dan etanol tidak toksik terhadap sel Vero. Fraksi n-heksana, etil asetat, dan air dari tidak toksik terhadap sel Vero.
Kata kunci: Alamar blue, ekstrak, fraksi, Punica granatum, sel Vero, Syzygium aqueum.
Syzygium aqueum and Punica granatum are plants commonly found in Indonesia. Flavonoids from Syzygium aqueum leaves and Punica granatum peels are known to have antioxidant effect for the human body. Because of that these plants widely used for treatment several diseases. Traditionally, Syzygium aqueum leaves and Punica granatum peels are commonly used as antibiotic, antidiabetic, and treatment for diarrhea. This study aim to determine in vitro toxicity effect of Syzygium aqueum and Punica granatum extracts and fractions using Vero cell lines. Crude drug of Syzygium aqueum was extracted by maceration, while crude drug of Punica granatum was extracted by reflux. Both crude drugs were extracted using continuous extraction method and solvents with increasing polarity which were n-hexane, ethyl acetate, and ethanol. Extracts were monitored by thin layer chromatography (TLC). The ethanol extract from Syzygium aqueum and Punica granatum were fractionated by liquid-liquid extraction using n-hexane, and ethyl acetate as solvents. Fractions were monitored by thin layer chromatography (TLC). Toxicity effect of Syzygium aqueum and Punica granatum extracts and fractions on Vero cells were evaluated using Alamar blue. Extracts and fractions of Syzygium aqueum leaves and Punica granatum had no toxicity effect against Vero cell at concentrations between 2 g/ml-1024 g/ml. Based on our findings all of extracts and fractions had no toxicity effect against Vero cell lines.
Keyword: Alamar blue, extract, fraction, Punica granatum, Syzygium aqueum, Vero cell.
References
Aulia AN, 2016, Pengujian toksisitas ekstrak keong matah (Cerithidea obtusa) terhadap Artemisia salina dan sel Vero, tugas akhir, Fakultas Perikanan dan Ilmu Kelautan IPB, Bogor.
Eisenbrand G, Pool-Zobel B, Baker V, Balls M, Blaauboer BJ, Boobis A, Carere A, Kevekordes S, Lhuguenot JC, Pieters R, Kleiner J, 2002, Method of in vitro toxicology, Food and Chemical Toxicology 40: 193-236.
Ekwall B, Silano V, Paganuzzi SA, Zucco F, Bourdeau (edd), 1990, Toxicity with mammalian cell cultures, John Willey and Sons Ltd, London.
Jahromi SB, Pourshafie MR, Mirabzadeh E, Tavasoli A, Katiraee F, Mostafavi E, Abbasian S, 2014, Punica granatum peel extract toxicity in mice, Jundishapur Journal of Natural Pharmaceutical Products 10(4): e23770, doi: 10.17795/jjnpp-23770.
Ling LT, Palanisamy UD, 2012, Review: Potential antioxidant from tropical plants, Food Industrial Processes-Methods and Equipment, Valdez B (eds), ISBN: 978-953-307-905-9, In-Tech, Eorope.
McLaughlin JL, Rogers LL, 1998, The use of biological assay to evaluate botanicals, Drug Information Journals 32: 512-534.
Manaharan T, Cheng MJ, Ming H, Uma D, 2013, Syzygium aqueum leaf extract and its bioactive compound enhance pre-adipocyte differentiation and 2 NBDG uptake in 3-T31 cell. Food Chemistry: 136-354-36.
Manaharan T, Chakravarthi S, Radhakrishnan AK, Palanisamy UD, 2014, In vivo toxicity evaluation of a standardized extract of Syzygium aquem leaf, Toxicology report 1: 718-725, doi: 10.1016/j.toxrep.2014.09.006.
Ramadania ZM, 2015, Isolasi senyawa 5,7-dihidroksi, 6,8-dimetil flavon dan uji aktivitas antioksidan serta penghambatan enzim xanthin oksidase dari ekstrak daun jambu air, tugas akhir, Sekolah Farmasi ITB, Bandung.
Patel S, Gheewala N, Suthar A, Shah A, 2009, In vitro cytotoxicity activity of Solanum nigrum extract against Hela cell line and Vero cell line, International Jornal of Pharmacy and Pharmaceutical Science vol 1, suppl 1, Nov-Dec 2009.
Ricardo CB, Monica AL, Sonia MG, Fabiana MB, Priscilia MA, 2009, A simple method to measure cell viability in proliferation and cytotoxicity assays, Braz Oral Res 23(3): 255-262.
RS Arvind Bharani, Selvaraj K, 2016, Pomegranate (Punica Granatum L.) peel extract- a study on potential sources of pharmacological activities, International Journal Pharmacy Bioscience: 282-290, doi: 10.22376/ljpbs.2016.7.4.b282-290.
