BAB I PENDAHULUAN 1.1 Latar Belakang Masalah Pisang (Musa Parasidiaca) merupakan tanaman buah – buah – buahan buahan tropika yang berasal dari Asia tenggara, Brazil dan India. I ndia. Di Asia Tenggara, pisang diyakini berasal dari Semenanjung Malaysia dan Filipina. Pisanh telah lama berkembang di India yaitu sejak 500 tahun sebelum masehi mase hi dan menyebar sampai ke daerah Pasifik. Pisang memiliki peranan penting di Indonesia karena dikonsumsi oleh konsumen tanpa memperhatikan tingkat sosial (Satuhu dan Supriadi, 2000). Indonesia merupakan salah satu sentra primer keragaman pisang, baik pisang segar, olahan dan pisang liar. Lebih dari 200 jenis pisang terdapat di Indonesia. Sentra produksi produksi pisang di Indonesia tersebar di 16 provinsi, 70 kabupaten. Provinsi tersebut antra lain NAD, Sumatera Utara, Sumatera Barat, Sumatera Selatan, Lampung, Riang, Riang, Jawa timur, Jawa Barat, Jawa Tengah, Banten, Bali, Kalimantan Barat, Kalimantan Selatan, Kalimantan Timur, Sulawesi Selatan, dan Maluku Utara. Selama periode 1995 sampai 2002 luas panen pisang berfluktuasi, namun pada tahun 2003 – 2003 – 2004 2004 cenderung meningkat (BPS, 2003). Di Indonesia tanaman pisang adalah tanaman yang multiguna, selain buahnya yang digunakan sebagai bahan konsumsi, daunnya juga dapat digunakan sebagai pembungkus dan bakal buahnya atau yang serinh dikenal sebagai jantung pisang digunakan sebagai sayur. Pisang memiliki kandungan gizi seperti karbohidrat, vitamin, mineral, air, lemak dan protein (Direktor Jendral Bina Reproduksi Hortikultura, 2003). Selain itu, pisang merupakan jenis buah yang mengandung banyak senyawa kimia yang bersifat antioksidan. Penelitian terhadao pisang menunjukan bahwa pisang tersebut banyak mengandung phenolik serta karotene (Fatemeh et al., 2012). Selain pada buah pisang, antioksidan juga terdapat pada kulit pisang. Antioksidan yang terdapat pada kulit pisang
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memiliki aktivitas yang lebih tinggi dibandingkan dengan buah pisang sendiri (Nagabhushan dan Bhide, 1998). Pemanfaatan kulit pisang di Indonesia,terbatas sebagai campuran pakan ternak (20-30%), serta serta pupuk kandang dan kompos kompos (60-70%) (Husni, 2009). Eksplorasi potensi kulit pisang pada bidang kesehatan be lum banyak dilakukan, meskipun beberapa peneliti melaporkan bahwa kulit pisang mengandung nutrien penting bagi kesehatan yang tidak kalah dengan dengan daging buahnya. Fokus penelitian terdahulu lebih banyak pada karakterisasi kandungan nutrien kulit pisang, aktivitas ekstrak kulit pisang sebagai antimikrobia, dan antibiotik alami, sedangkan aktivitas ant ioksidan pada kulit pisang belum belum banyak diteliti (Mokbel and Hashinaga, 2005). Aktivitas antioksidan dan antimikrobial pada kulit pisang terjadi karena pada kulit pisang juga terkandung senyawa organic seperti pada bagian tanaman pisang yang yang lain. Senyawa Senyawa organic yang terdapat pada kulit kulit pisang merupakan jenis senyawa golongan flavonoid seperti sianidin, delpinidin, petunidin, dan malvidin-3-ramnosil-1,6-glukosid malvidin-3-ramnosil-1,6-glukosida. a.
Gambar 1 Senyawa Metabolit Sekunder Dalam Kulit Pisang
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Pada penelitian ini akan dilakukan pemanfaatan limbah kulit pisang untuk didapatkan ekstrak senyawa bahan alamnya dan diuji dayanya dalam aplikasinya sebagai antioksidan.
1.2 Rumusan Masalah 1. Apa metode ekstraksi yang dapat digunakan untuk mendapatkan kandungan senyawa bahan alam yang terdapat pada kulit pisang secara optimal dan pelarut apa yang digunakan? 2. Apa saja senyawa bahan alam yang terkandung pada kulit pisang? 3. Bagaimana aktivitas antioksidan ekstrak kulit pisang? 4. Berapa nilai IC50 ekstrak kulit pisang? 5. Berapa nilai total kadar fenol dan total kadar flavonoid dari ekstrak kulit pisang?
1.3 Hipotesis 1. Metode ekstraksi yang dapat menyari kandungan bahan alam pada kulit pisang dengan maksimal adalah dengan menggunakan metode maserasi dengan menggunakan pelarut polar 2. Senyawa bahan alam yang terdapat pada kulit pisang adalah flavonoid dan saponin 3. Aktivitas antioksidan kulit pisang adalah cukup kuat 4. Nilai IC50 kulit pisang dibawah 100 5. Nilai total kadar fenol dan total kadar flavonoid dari kulit pisang kurang
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1.4 Tujuan Penelitian 1. Memanfaatkan limbah kulit pisang yang selama ini disia-s iakan 2. Mengetahui kondisi yang optimum untuk mengekstrak kulit pisang 3. Memanfaatkan ekstrak kulit pisang sebagai penangkal radikal bebas
1.5 Manfaat Penelitian Hasil yang diperoleh dari penelitian ini adalah untuk menambah inventaris ekstrak senyawa bahan alam yang dapat digunakan untuk menangkal radikal bebas.
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BAB II TINJAUAN PUSTAKA 2.1 Pisang
Pisang ( Musa paradisiac) paradisiac) adalah tanaman buah yang berupa herba yang banyak terdapat di Asia Tenggara, seperti Indonesia, dan terdapat juga di Afrika, Amerika Tengah, dan Amerika Selatan. Tanaman ini mengandung mengandung karbohidrat, vitamin, vitamin, dan mineral, terutama kalium. Selain Se lain itu, tanaman pisang juga mengandung beberapa senyawa organic, seperti saponin, tanin, dan flavonoid. Beberapa manfaat dari tanaman pisang antara lain:
Mempercepat penyembuhan luka
Mengatasi jerawat
Meredakan nyeri Kingdom: Plantae Divisi : Spermatophyta Sub Divisi : Angiospermae Kelas : Monocotyledonae Famili : Musaceae Genus : Musa Spesies : Musa spp.
Gambar 2 Buah Pisang
Kulit pisang adalah salah satu bagian yang terdapat dari tanama pisang. Kulit Kulit pisang ini membungkus buah pisang, dan biasanya hanya menjadi limbah saja. Kalaupun dimanfaatkan hanya sebatas untuk pakan ternak. Padahal, di dalam kulit pisang terdapat senyawa flavonoid yang cukup banyak yang dapat dimanfaatkan. Selain flavonoid, pada kulit pisang juga terdapat senyawa saponin. Sama seperti halnya ha lnya buahnya, kulit pisang
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2.2 Ekstraksi
Ekstraksi adalah penyarian zat-zat aktif dari bagian tanaman obat. Adapun tujuan dari ekstraksi yaitu untuk menarik komponen kimia yang terdapat dalam simplisia. Tujuan ekstraksi adalah untuk menarik semua komponen kimia yang terdapat dalam simplisia. Ekstraksi ini didasarkan pada perpindahan massa komponen zat padat ke dalam pelarut dimana perpindahan mulai terjadi pada lapisan antar muka, kemudian berdifusi berdifusi masuk ke dalam pelarut. Ekstrak adalah sediaan kering, kental, atau cair dibuat dengan menyaring simplisia nabati dan hewani menurut cara yang cocok, diluar pengaruh matahari yang langsung. Ekstrak kering harus lebih mudah digerus menjadi serbuk. Terdapat beberapa jenis ekstrak baik ditinjau dari segi pelarut yang digunakan ataupun hasil akhir dari ekstrak tersebut (4). 1) Ekstrak air Menggunakan pelarut air sebagai cairan pengekstraksi. Pelarut air merupakan pelarut yang mayoritas digunakan dalam proses ekstraksi. Ekstrak yang dihasilkan dapat langsung digunakan atau diproses kembali seperti melalui pemekatan atau proses pengeringan . 2) Tinktur Sediaan cari yang dibuat dengan cara maserasai ataupun perkolasi simplisia. Pelarut yang umum digunakan dalam proses produksi tinktur adalah etanol. Satu bagian simplisia diekstrak dengan menggunakan 2-10 bagian menstrum/ekstraktan. 3) Ekstrak cair Bentuk dari ekstrak cair mirip dengan tinktur namun telah melalui pemekatan hingga diperoleh ekstrak yang sesuai dengan ketentuan farmakope.
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5) Ekstrak kental Ekstrak ini merupakan ekstrak yang telah mengalami proses pemekatan. Ekstrak kental sangat mudah untuk menyerap menyerap lembab sehingga mudah untuk ditumbuhi oleh kapang. Pada proses industri ekstrak kental sudah tidak lagi digunakan, hanya merupakan tahap perantara sebelum diproses kembali menjadi ekstrak kering. 6) Ekstrak kering (extract sicca) Ekstrak kering merupakan ekstrak hasil pemekatan yang kemudian dilanjutkan ke tahap pengeringan. Proses pengeringan dapat dilakukan dengan berbagai macam cara yaitu dengan menggunakan bahan tambahan seperti laktosa atau aerosil, menggunakan proses kering beku namun proses ini tidak ekonomis, dan dengan menggunakan proses semprot kering atau fluid bed drying . 7) Ekstrak minyak Dilakukan
dengan
cara
mensuspensikan
simplisia
dengan
perbandingan tertentu ter tentu dalam minyak yang telah t elah dikeringkan, dengan cara seperti maserasi. 8) Oleoresin Merupakan sediaan yang dibuat dengan cara ekstr aksi bahan oleoresin (mis. Capsicum fructus dan zingiberis rhizom) dengan pelarut tertetu umumnya etanol. Terdapat beberapa istilah yang perlu diketahui berkaitan dengan proses ekstraksi adalah ekstraktan/menstrum ekstraktan/menstrum yaitu pelarut/campuran pelarut pelarut
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d. Jenis pelarut yang digunakan e. Jenis pelarut berkaitan dengan polaritas dari pelarut tersebut. Hal yang perlu diperhatikan dalam proses ekstraksi adalah senyawa yang memiliki kepolaran yang sama akan lebih mudah tertarik/ tert arik/ terlarut dengan pelarut yang memiliki tingkat kepolaran yang sama. Berkaitan dengan polaritas dari pelarut, terdapat tiga golongan pelarut yaitu: a) Pelarut polar Memiliki tingkat kepolaran yang tinggi, cocok untuk mengekstrak senyawa-senyawa yang polar dari tanaman. Pelarut polar cenderung universal digunakan karena biasanya walaupun polar, tetap dapat menyari senyawa-senyawa dengan tingkat kepolaran lebih rendah. Salah satu contoh pelarut polar adalah: air, metanol, etanol, asam asetat. b) Pelarut semipolar Pelarut semipolar memiliki tingkat kepolaran yang lebih rendah dibandingkan dengan pelarut polar. Pelarut ini baik untuk mendapatkan senyawa-senyawa semipolar dari tumbuhan. Contoh pelarut ini adalah: aseton, etil asetat, kloroform. c) Pelarut nonpolar Pelarut nonpolar, hampir sama sekali tidak polar. Pelarut ini baik untuk mengekstrak senyawa-senyawa yang sama sekali tidak larut dalam pelarut polar. Senyawa ini baik untuk mengekstrak mengekstrak berbagai jenis minyak. Contoh: heksana, eter.
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1. Lama waktu ekstraksi Lama ekstraksi akan menentukan banyaknya senyawa-senyawa yang terambil. Ada waktu saat pelarut/ ekstraktan jenuh. Sehingga tidak pasti, semakin lama ekstraksi semakin bertambah banyak ekstrak yang didapatkan. 2. Metode ekstraksi, termasuk t ermasuk suhu yang digunakan. Terdapat banyak metode ekstraksi. Namun secara ringkas dapat dibagi berdasarkan suhu suhu yaitu metode metode ekstraksi dengan dengan cara panas panas dan dan cara dingin. dingin. Metode panas digunakan jika senyawa-senyawa yang terkandung sudah dipastikan tahan panas. Proses pengekstraksian komponen kimia dalam sel tanaman yaitu pelarut organik akan menembus dinding dinding sel dan masuk masuk ke dalam rongga sel yang mengandung zat aktif, aktif, zat aktif akan larut dalam pelarut organik di luar sel, maka larutan terpekat akan berdifusi keluar sel dan proses ini akan berulang terus sampai terjadi keseimbangan antara konsentrasi cairan zat aktif di dalam dan di luar sel.
2.3 Maserasi
Maserasi atau macerare(Bahasa macerare(Bahasa Latin, artinya merendam) adalah ekstraksi bahan nabati dengan cara merendam simplisia kedalam suatu pelarut, baik polar maupun non polar, selama beberapa waktu sambil sesekali diaduk. Prinsip dari ekstraksi dengan cara maserasi adalah difusi senyawa yang terdapat pada simplisia dengan pelarut sehingga tercapai
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4. Tanpa pemanasan Meskipun metode ekstraksi dengan maserasi memiliki keuntungan dalam hal kemudahan untuk melakukannya, metode ini memiliki ke lemahan seperti tidak terekstraknya seluruh senyawa simplisia (50%) dan waktu yang relative lama.
Gambar 3 Prinsip Kerja Ekstraksi Metode Maserasi
2.4 Flavonoid
Flavonoid merupakan molekul polifenol yang larut dalam air yang
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Flavonoid merupakan senyawa polifenol yang melimpah di alam dan dikategorikan menurut struktur kimianya ke dalam flavonol, flavon, flavanone, isoflavon, katekin, antosianidin, dan kalkon (Tanaka, et al , 2008). Pentingnya senyawa polifenol terhadap kesehatan manusia telah dipelajari secara massif dalam beberapa tahun terakhir, khususnya golongan flavonoid. Flavonoid dapat berguna sebagai antimikroba, fotoreseptor, antioksidan, antiallergenic, dan anti inflamasi.
Gambar 5 Golongan Senyawa Flavonoid
2.5 Senyawa Fenolik
Senyawa fenolik fenolik merupakan senyawa yang banyak ditemukan ditemukan pada tumbuhan. Fenolik memiliki cincin aromatik satu atau lebih gugus hidroksi
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di semua tanaman, daun, bunga dan buah. Ribuan senyawa fenolik alam telah diketahui strukturnya,antara lain flavonoid, fenol monosiklik sederhana, fenil propanoid, polifenol (lignin, melanin, tannin), dan kuinon fenolik.
Banyak senyawa fenolik alami mengandung sekurang-kurangnya satu gugus hidroksil hidroksil dan lebih banyak yang membentuk senyawa eter, ester atau glioksida daripada senyawa bebasnya. Senyawa ester atau eter fenol tersebut memiliki kelarutan yang lebih besar dalam air daripada senyawa fenol dan senyawa glioksidanya.
Dalam keadaan murni, senyawa fenol berupa zat padat yang tidak berwarna, tetapi jika teroksidasi akan berubah menjadi gelap. Kelarutan fenol dalam air akan bertambah, jika gugus hidroksil makin banyak. Senyawa Senyawa fenolik memiliki aktivitas biologis yang beraneka ragam, dan banyak digunakan dalam reaksi enzimatik oksidasi kopling sebagai substrat donor H. Reaksi oksidasi kopling, selain membutuhkan suatu
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aroma atau warna pada tumbuhan itu. Senyawa kimia tidak termasuk ke dalam zat gizi karena bukan berupa karbohidrat, protein, lemak, vitamin, mineral maupun air. Dalam penggunaan umum, fitokimia memiliki definisi yang lebih sempit. Fitokimia biasanya digunakan untuk merujuk pada senyawa yang ditemukan pada tumbuhan yang tidak dibutuhkan untuk fungsi normal tubuh, tapi memiliki efek yang menguntungkan bagi kesehatan atau memiliki peran aktif bagi pencegahan penyakit. Fitokimia, senyawa yang begitu bermanfaat sebagai antioksidan dan mencegah kanker juga penyakit penyakit jantung. Beberapa studi pada manusia dan hewan membuktikan zat – zat kombinasi fitokimia ini didalam tubuh memiliki fungsi tertentu yang berguna bagi kesehatan. Kombinasi Kombinasi itu antara lain menghasilkan menghasilkan enzim – enzim sebagai penangkal racun, merangsang sistem pertahanan tubuh, mencegah penggupalan keeping – keeping darah, menghambat sintesa kolesterol dihati, dan meningkatkan metabolisme hormon. Secara garis besar fitokimia terdiri ter diri dari alkaloid, flavonoid, terpenoid, steroid, stero id, saponin,
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2.6.2 Flavonoid
Flavonoid adalah kelompok senyawa fenol terbesar yang ditemukan di alam terutama pada jaringan jaringan tumbuhan tinggi. Senyawa Senyawa ini merupakan produk metabolik sekunder yang terjadi dari sel dan terakumulasi dari tubuh tumbuhan sebagai zat racun ( Robinson, 1991). Senyawa flavonoid mempunyai mempunyai kerangka dasar karbon dalam inti int i dasarnya yang tersusun dalam konfigurasi C 6 - C3 – C6. Susunan tersebut dapat menghasilkan tiga struktur yaitu: 1,3-diarilpropana (flavonoid),
1,2-diarilpropana
(isoflavonoid),
2,2-diarilpropana
(neoflavonoid). Menurut Markham (1982), flavonoid merupakan senyawa polar karena mempunyai gugus hidroksil yang tak tersulih, atau suatu gula, sehingga flavonoid cukup larut dalam pelarut polar seperti etanol, metanol, butanol dan air. Flavonoid umumnya terikat pada gula sebagai glukosida dan aglikon flavonoid. Uji warna yang penting dalam larutan alkohol ialah direduksi dengan serbuk Mg dan HCl pekat. Diantara
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a.
Monoterpen terdiri dari dua unit C5 atau 10 atam karbon.
b.
Siskuisterpen terdiri dari tiga unit unit C 5 atau 15 atom ato m karbon
c.
Diterpen terdiri dari empat unit C5 atau 20 atom karbon
d.
Triterpen terdiri dari enam unit C5 atau 30 atom karbon
e.
Tetraterpen terdiri dari delapan unit C5 atau 40 atom karbon
Secara kimia, terpenoid umumnya larut dalam lemak dan terdapat didalam sitoplasma sel tumbuhan. Biasanya diekstraksi memakai petrolium
eter,
eter
atau
kloroform
dan
dapat
dipisahkan secara kromatografi pada silika gel dengan pelarut ini (Harborne,1987). Steroid adalah terpenoid yang kerangka dasarnya terbentuk dari sistem cincin siklopentana prehidrofenantrena. Steroid merupakan golongan senyawa metabolik sekunder yang banyak dimanfaatkan sebagai obat. Hormon steroid pada umumnya diperoleh dari senyawasenyawa steroid alam terutama dalam tumbuh tu mbuhan an (Djamal, 1988).
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juga digunakan secaa luas dalam indutri makanan, industry industry petroleum, industry karet dan sebaginya sebaginya (Tahir, Wijaya, dan Widyaningsih, Widyaningsih, 2003). 2003). Antioksidan dapat bersumber dari zat – zat zat alami hasil isolasi. Anya antioksidan alami maupun sintesis dapat menhambat oksidasi lipid, mencegah kerusakan perubahann degradsi komponen organic dalam bahan makanan. Beberapa senyawa antioksidan sintesis yang umum digunakan adlah butylated hydroxytoluen (BHT), butylated hydroxyanisole (BHA), terbutylhydroxyquinone (TBHQ), asam galat dan propil galat. Antioksidan alami dapat diperoleh dari makanan sehari – hari hari seperti sayuran, buah – buahan, kacang – kacangan dan tanaman lainnya yang mengandung antioksidan bervitamin seperti vitamin A, C dan E, asam-asam fenolat seperti asam ferulat, asam klorogerat, asam elegat, dan asam kafeat, dan senyawa flavonoid seperti kuersetin, mirisetin, apigenin, luteolin, dan kaemferol (Rohdiana, 2001, Pokornya et al , 2001).
2.8 Uji Aktivitas Antioksidan
Untuk menentukan aktivitas antioksidan metode yang dilakukan
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2.9 Spektr Spektrofotometer ofotometer UV-Visible
Spektrofotometri Sinar Tampak (UV-Vis) adalah pengukuran energi cahaya oleh suatu systempada s ystempada panjang gelombang tertentu. Sinar ultraviolet (UV) mempunyai panjang gelombang antara 200-400 nm, dan sinar t ampak (visible) mempunyai panjang gelombang 400-750 nm. Pengukuran spektrofotometri menggunakan alat spektrofotometer yang melibatkan energi elektronik yang cukup besar pada molekul yang dianalisis, sehingga spektrofotometer UV-Vis lebih banyak dipakai untuk analisis kuantitatif dibandingkan
kualitatif.
Spektrum
UV-Vis
sangat
berguna
untuk
pengukuran secara kuantitatif. Konsentrasi dari analit d i dalam larutan bisa
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Indeks bias tidak tergantung pada konsentrasi larutan
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2.9.1.2
Wadah Sampel
Kebanyakan spektrofotometri melibatkan larutan dan karenanyan kebanyakan wadah sampel adalah sel/kuvet untuk menaruh cairan ke dalam berkas cahaya spektrofotometer. Kuvet itu harus dapat meneruskan energi cahaya dalam daerah spektral yang diminati, jadi kuvet kaca melayani daerah tampak, kuvet kuarsa atau silica untuk daerah ultraviolet.
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2.9.1.4
Detektor
Detektor akan menangkap sinar yang diteruskan oleh larutan. Sinar kemudian diubah menjadi sinyal listrik oleh amplifier dan dalam rekorder dan ditampilkan dalam bentuk angka-angka padakomputer.
2.9.1.5 Recorder/Visual Display Merupakan system baca yang memperagakan besarnya isyarat listrik, menyatakan dalam bentuk % Transmitan maupun Absorbansi.
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BAB III METODOLOGI PRAKTIKUM 3.1 Waktu dan Tempat Pelaksanaan Penelitian dilakukan pada bulan September-Oktober 2014 bertempat di Pusat Laboratorium Terpadu UIN Syarif Hidayatullah Jakarta
3.2 Alat dan Bahan 3.2.1 Ekstraksi Maserasi Alat yang digunakan antara lain blender, Erlenmeyer 250mL, neraca analitik, gelas beker 100mL, spatula, gelas ukur 100mL, rotary evaporator, kaca arloji, oven, corong, dan desikator. Bahan yang
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3.2.4 Uji Kadar Total Fenol dan Flavonoid Alat yang digunakan pada praktikum ini adalah tabung reaksi, rak tabung reaksi, pipet gondok, pipet tetes, gelas ukur, dan labu ukur, batang pengaduk, neraca timbang, spektrofotometer uv-vis. Bahan bahan yang yang digunakan digunakan untuk uji uji kadar total fenol adalah larutan Standar
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3.3.2.2
Uji Flavonoid
Ekstrak tanaman sebanyak 2 mL dimasukkan ke dalam tabung reaksi, kemudian ditambahkan sedikit serbuk Mg ke dalam tabung reaksi dan 1 mL HCl 2% (positif flavonoid jika timbul busa dan berwarna bening-orange.
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dikocok dengan kuat sampai timbul busa (positif saponin jika busa tersebut stabil selama 10 menit).
3.3.3 Uji Aktivitas Antioksidan Ekstrak tanaman kulit pisang hasil pemekatan ditimbang
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3.3.4.2
Kadar Total Flavonoid
Sebanyak 1 ml sampel dimasukkan ke dalam tabung reaksi dan ditambahkan 5 ml aquades. Lalu dipipet tepat 0,3 ml NaNO3 5% dan ditambahkan ke dalam tabung reaksi. Setelah itu, ditambahkan lagi 0,3 ml AlCl3 10%. Lalu disentrifuge selama 5 menit dan diinkubasi.
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BAB IV HASIL DAN PEMBAHASAN 4.1 Ekstraksi Maserasi Ekstraksi sampel kering kulit pisang menggunakan metode maserasi bertujuan untuk mendapatkan senyawa pada kulit ku lit pisang p isang yang tidak tahan t ahan
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