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Download This PDF File Prosiding Farmasi ISSN: 2460-6472 Uji Aktivitas Sitotoksik Ekstrak Etanol Kulit Buah Petai (Parkia Speciosa Hassk.) dengan Metode BSLT (Brine Shrimp Lethality Test) Cytotoxic Activity Test of Petai Fruit Peel (Parkia Speciosa Hassk.) Ethanolic Extract Using BSLT (Brine Shrimp Lethality Test) Method 1Rahayu Rahmadini Soliha, 2Sri Peni Fitrianingsih, 3Siti Hazar 1,2,3Prodi Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Islam Bandung, Jl. Tamansari No.1 Bandung 40116 email: [email protected], [email protected], [email protected] Abstract. Cancer is the third leading cause of global death globally, which continue to increase each year. Petai fruit peel (Parkia speciosa Hassk.) contains secondary metabolites that can be used as anticancer. This research was conducted to determine the secondary metabolites contents of petai fruit peel and to determine its cytotoxic activity as well as to calculate lethal concentration (LC50) of petai fruit peel ethanolic extract on shrimp larvae Artemia franciscana Kellogg. using Brine Shrimp Lethality Test (BSLT) method. LC50 value was calculated using probit analysis method. Extract concentrations that used in this cytotoxic test were 1, 10, 100, 200, 300, 400, 500, 600, 700, 800, 900, and 1000 ppm. Phytochemical screening showed positive results of flavonoids, saponins, tannins, polyphenolates, alkaloids, steroids, triterpenoids, monoterpenes and sesquiterpenes. From cytotoxicity test, it was found that LC50 of the extract against shrimp larvae Artemia fraciscana Kellogg was 202.4417 ppm, which ranged in moderate category with the value between 100 to 1000 ppm. Keywords: Cytotoxic test, Petai Fruit peel , Artemia franciscana Kellogg., Brine Shrimp Lethality Test (BSLT) methode Abstrak. Penyakit Kanker merupakan penyebab kematian ketiga di dunia yang terus meningkat setiap tahun. Kulit buah petai (Parkia speciosa Hassk.) diduga mengandung senyawa metabolit sekunder yang dapat digunakan sebagai antikanker. Penelitian ini dilakukan dengan tujuan untuk mengetahui kandungan metabolit sekunder dari kulit buah petai (Parkia speciosa Hassk.) dan mengetahui aktivitas sitotoksik serta nilai Lethal Concentration (LC50) dari ekstrak etanol kulit buah petai (Parkia speciosa Hassk.) terhadap larva udang Artemia franciscana Kellogg. dengan metode Brine Shrimp Lethality Test (BSLT). Nilai LC50 dihitung menggunakan metode analisis probit. Konsentrasi ekstrak etanol yang digunakan pada pengujian sitotoksik ini adalah 1; 10; 100; 200; 300; 400; 500; 600; 700; 800; 900 dan 1000 ppm. Pada penapisan fitokimia yang terdeteksi adalah flavonoid, saponin, tanin, polifenolat, alkaloid, steroid, triterpenoid serta monoterpen dan seskuiterpen. Pada pengujian sitotoksik diperoleh nilai LC50 ekstrak etanol kulit buah petai adalah 202,4417 ppm. Berdasarkan data tersebut, ekstrak etanol 96% dari kulit buah petai menyebabkan sitotoksik terhadap larva udang Artemia fraciscana Kellogg. yang termasuk kedalam kategori moderat karena memiliki nilai LC50 antara 100 sampai 1000 ppm. Kata kunci: Uji sitotoksik, kulit buah petai, Artemia franciscana, metode Brine Shrimp Lethality Test (BSLT) A. Pendahuluan penyakit yang sukar disembuhkan dan Penyakit kanker merupakan dapat menyebabkan kematian, suatu penyakit yang disebabkan oleh sehingga hal ini merupakan masalah pertumbuhan sel-sel jaringan tubuh yang sulit dalam bidang pengobatan tidak normal, ditandai dengan (Yudistira, 2017:46). pembelahan sel tidak terkendali dan Pemanfaatan tumbuhan alam kemampuan sel-sel tersebut menyerang sebagai alternatif terapi kanker saat ini jaringan biologis (Katzung, 2012:907) sudah mulai berkembang karena terapi Penyakit kanker dikenal sebagai kanker menggunakan obat kimia 566 Uji Aktivitas Sitotoksik Ekstrak Etanol Kulit Buah Petai... | 567 umumnya belum mampu memberikan Kingdom : Plantae hasil yang memuaskan dan mempunyai Divisi : Magnoliophyta biaya yang mahal, selain itu ditemukan Kelas : Magnoliopsida efek samping yang cukup besar, salah Ordo : Fabales satunya adalah kulit buah petai (Parkia Famili : Fabaceae speciosa Hassk.). Chanu et al., (2018) Genus : Parkia mengemukakan bahwa biji tanaman Spesies : Parkia speciosa Hassk (Parkia javanica) dapat menghambat Petai (Parkia speciosa Hassk.), proliferasi sel kanker hati manusia merupakan tanaman bermanfaat yang HepG2. Berdasarkan kesamaan genus, tumbuh di daerah tropis dan umum maka diduga petai dari spesies Parkia dikonsumsi di Indonesia yang banyak speciosa Hassk juga memiliki potensi digunakan oleh masyarakat sebagai sebagai antikanker. Selain itu kulit buah bahan pangan dan industri, serta petai juga mengandung senyawa berpotensi dimanfaatkan sebagai obat- metabolit sekunder flavonoid, saponin, obatan seperti obat hati, ginjal, tannin dan triterpenoid yang diketahui sembelit, depresi, dan anemia berpotensi sebagai antikanker. (Zulhendra et al., 2016:102). Untuk mengetahui aktivitas kulit Kulit buah petai (Parkia buah petai (Parkia speciosa Hassk.) spesiosa Hassk.) mengandung berbagai sebagai agen antikanker dilakukan uji macam senyawa metabolit sekunder. sitotoksik dengan metode Brine Shrimp Pada penelitian Jebarus (2015) ekstrak Lethality Test (BSLT) menggunakan etanol kulit buah petai dapat berpotensi larva udang (Artemia) dengan parameter sebagai antibakteri dengan kandungan nilai LC50 (Meyer et at., 1982:32). senyawa saponin, flavonoid, alkaloid, Berdasarkan penjelasan tersebut, tanin, polifenolat, triterpenoid, steriud, dapat dirumuskan masalah yaitu, apa monoterpen dan seskuiterpen. saja senyawa metabolit sekunder yang Uji sitotoksik merupakan uji terkandung dalam ekstrak etanol kulit invitro dengan menggunakan kultur sel buah petai (Parkia speciosa Hassk.) untuk mendeteksi adanya aktivitas sehingga memiliki aktivitas sitotoksik antikanker dari suatu senyawa. Salah terhadap Artemia franciscana Kellogg. satu uji sitotoksik adalah metode Brine dan berapakah nilai LC50 ekstrak etanol Shrimp Lethality Test (BSLT) dengan kulit buah petai menggunakan larva udang (Artemia Tujuan dari penelitian ini adalah franciscana Kellogg.). Metode ini untuk mengetahui kandungan senyawa merupakan penapisan awal dalam upaya metabolit sekunder ekstrak etanol kulit pencarian senyawa antikanker karena buah petai (Parkia speciosa Hassk.) hasil dari uji tersebut memiliki korelasi serta aktivitas sitotoksik terhadap larva positif dengan aktivitas sitotoksik udang (Artemia franciscana Kellogg.) antikanker. Keuntungan metode ini sehingga dapat dilihat potensinya adalah cepat, tidak mahal, tidak sebagai antikanker dan untuk membutuhkan peralatan yang rumit, mengetahui nilai LC50 ekstrak etanol mudah dilakukan, selain itu metode kulit buah petai. BSLT sering digunakan dalam tahap B. Landasan Teori awal isolasi senyawa toksik yang terkandung didalam suatu ekstrak Klasifikasi tanaman petai dengan tingkat kepercayaan 95% menurut Cronquist (1981) adalah sehingga hasilnya dapat dipercaya Data sebagai berikut: yang diperoleh diolah untuk Farmasi 568 | Rahayu Rahmadini Soliha, et al. mendapatkan nilai LC50 (Lethal Institut Teknologi Bandung dan larva Concentration 50) yang merupakan udang di Museum Zoologi, Sekolah Ilmu konsentrasi zat yang menyebabkan dan Teknologi Hayati, Institut Teknologi terjadinya kematian pada 50 % hewan Bandung percobaan yaitu larva udang (Artemia Tahap selanjutnya pembuatan franciscana) (Meyer et al., 1982). simplisia yaitu sortasi basah, pencucian Tabel 1. Klasifikasi sitotoksik untuk dan pengeringan. Simplisia kemudian bahan alam menurut Balantyne dalam dikarakterisasi dengan menguji Mardja (2016) parameter standar spesifik dan parameter standar nonspesifik. Untuk mengetahui kandungan senyawa metabolit sekunder, LC50 Kategori simplisia dan ekstrak diuji penapisan 10μg/mL< LC50 Sangat toksik fitokimia. Selanjutnya simplisia 10μg/mL<LC50<100μg/mL Toksik diekstraksi menggunakan metode 100μg/mL<LC50<1000μg/mL Moderat maserasi dengan pelarut etanol 96% dan LC50>1000μg/mL Tidak toksik dipekatkan dengan cara evaporasi. Pengujian sitotoksik ekstrak Artemia atau brine shrimp etanol kulit buah petai menggunakan merupakan golongan zooplankton yang metode BSLT (Brine Shrimp Lethality hidup sebagai planktonic, yaitu Test) terhadap larva udang dilakukan melayang dalam air. Artemia termasuk dengan beberapa variasi konsentrasi, jenis udang- udangan yang mempunyai dimana tiap konsentrasi dilakukan ukuran relatif kecil dengan sistem pengulangan sebanyak 3 kali (triplo) osmoregulasi yang efisien sehingga dengan parameter yang diuji adalah mampu beradaptasi pada kisaran persen kematian untuk menghitung nilai salinitas yang luas (5-150 ppt). Faktor LC50. Nilai LC50 yang diperoleh lingkungan yang optimal untuk menunjukan tingkat aktivitas sampel kehidupan artemia adalah pada suhu 25- yang diuji. 40C; pH berkisar 7,3-8,3; serta oksigen terlarut lebih dari 2 mg/L (Wibowo, D. Hasil Penelitian dan 2013: 3). Pembahasan C. Metodologi Penelitian Hasil determinasi menunjukkan bahwa tanaman petai yang digunakan Penelitian ini bertujuan untuk adalah petai jenis Parkia speciosa menguji aktivitas sitotoksik ekstrak Hassk. Sedangkan larva udang yang etanol kulit buah petai (Parkia speciosa digunakan adalah Artemia franciscan Hassk.) terhadap larva udang Artrmia Kellogg. franciscana Kellogg. yang dilakukan Pembuatan simplisia kulit buah melalui pengujian eksperimental di petai dari bahan basah seberat 1,5 kg laboratorium. Penelitian diawali dengan menghasilkan 500 g berupa simplisia pengumpulan bahan uji kulit buah petai kering. Kemudian simplisia tersebut yang diperoleh dari Majalengka,
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