PERCOBAAN 5 TEKNIK MASERASI PADA PENETAPAN KADAR TANNIN DALAM TEH
I.
TUJUAN
1. Memisahkan senyawa tannin dalam sampel teh dengan teknik maserasi 2. Menetapkan kadar senyawa tannin hasil pemisahan dengan teknik titrasi permanganometri II.
PRINSIP
Ekstraksi tannin dalam sampel teh dapat dilakukan dengan teknik maserasi (perendaman). Maserasi tersebut menggunakan pelarut air. Filtrat yang diperoleh dititar dengan KMnO 4 0,1N untuk mengetahui kadar tannin dalam teh. Metode penentuan kadar tanin dalam teh secara titrasi permanganometrimerupakan suatu metode yang disebut dengan metode Lowenthal-Procter. Metode ini didasari oksidasi fenolat oleh larutan kalium permanganat dengan adanya indigo carmine sebagai indikator redoks untuk menunjukkan titik akhir titrasi.
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III. REAKSI 1. Reaksi Standarisasi
Setengah reaksi redoks larutan KMnO4 denganH2C2O4 Oks:
C2O4-2
2CO2 + 2e-
Ion oksalat oksalat
Red:
(karbondioksida)
MnO4- + 8H+ + 5e-
Mn2+ + 4H2O (x2)
Ion Mangan II
(Ion permanganat) (Ion Hidrogen)
5C2O4-2
(Air)
10CO2 + 10e(Karbon Dioksida)
(Ion oksalat)
2MnO4- + 16H + + 10e-
(Ion Permanganat)
2Mn2+ + 8H2O
(Ion Hidrogen)
(Ion Mangan II)
5C2O4-2 + 2MnO4- + 16H+ (Ion Oksalat)
(x5)
(Ion Permanganat)
(Ion Hidrogen)
(Basset J 1994 : halaman 409) 1
2. Reaksi Tanin
(Harbone,J.B.1987 (Harbone,J.B.1987 : Halaman 107) 2
(Air)
10CO2 + 2Mn2+ + 8H2O (Karbon Dioksida)
(Ion Mangan II)
(Air)
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IV.
DASAR TEORI
Tanaman teh merupakan salah satu tanaman yang termasuk kedalam genus camellia yang banyak tersebar di wilayah kawasan asia tenggara. Teh sendiri merupakan minuman yang mengandung kafein, yang dibuat dengan cara menyeduh daun pucuk, atau tangkai daun yang dikeringkan dari tanaman camellia sinensis dengan penambahan air panas. Teh dapat dikelompokan menjadi 3 golongan yaitu teh hijau, teh hit am, teh oolong. Ketiga jenis teh te h ini mengandung polifenol yang berfungsi sebagai antioksidan yang mampu melindungi tubuh dari serangan radikal bebas. Bahkan potensi antioksidan teh tersebut lebikuat dibandngkan antioksidan dalam sayuran dan buah-buahan . Dimana kandungan senyawa folipenol yang berfungsi sebagai antioksidan yang dapat melindungi tubuh dari radikal bebas. Bahkan antioksidan the dianggap lebih kuat dibandingkan dengan antioksidan dalam sayuran dan buah-buahan dimana kandungan senyawa polifenol yang berfungsi pada sayur dan buah hanya sekitar 0,25 % dan komponen tersebut umumnya terdapat dalam keadaan terikat atau terkonjugasi dengan senyawa gula, sedangkan senyawa komposisi polifenol pada teh yang terdiri dari flavanol, flavandiol, flavanoida, dan asam-asam fenolat diperkirakan sekitar 30 % dari berat kering daun teh. Sehingga banyak peneliti membuktikan senyawa polifenol sebagai anti-kanker terutama kanker lambung, esophagus dan kulit. Bahkan polifenol pun mampu menurunkan kolesterol dan mencegah penggumpalan (winarsi, 2007)1 Tanin merupakan turunan fenol. Pada miskroskop, tanin biasanya tampak sebagai massa butiran bahan berwana kuning, merah atau coklat. Tanin dapat ditemukan dalam bagian yang berbeda dari tumbuhan, misalnya mi salnya pada daun, jaringan pembuluh, buah yang belum masak, kulit, biji, dan jaringan tumbuhan karena penyakit. Tanin dapat ditemukan dalam sel biasa atau dalam iodiblas. Didalam sel, tanin terdapat dalam vakuola atau dalam
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dan perusakan oleh hewan. Secara komersial, tanin digunakan sebagai khususnya dalam industri penyamakan kulit (mulyani, 2006)2 Tanin mengandung sebagian besar gugus hidroksifenolik. Proteksi dari serangan ternak dapat dilakukan dilakukan dengan menimbulkan rasa sepat, serangan dari bakteri dan insekta diproteksi dengan menonaktifkan enzim-enzim protoase dari bakteri dan insekta yang bersangkutan (Cheeke dan Shull, 1985)3. Tanin juga membentuk komplek dengan komponen polimer dinding sel dari serangan organisme patogen dan menghentikan pembelahan sel (Swain, 1979)4. Tanin secara umum dibagi menjadi dua kelas, yaitu : 1. Tanin Kondensasi Tanin kondensasi dikenal juga sebagai proanthocyanidin, adalah paling banyak tedistribusi pada tanaman, tidak mudah dihidrolisis dan terdapat dalam struktur yang komplek (Cheeke dan Shull, 1985). Tanin kondensasi merupakan senyawa polimer dari flavan -3-01 (catekin) atau flavan -3; 4-diol (leucoanthocyanidin) atau turunannya yang dihubungkan oleh ikatan C-C atau C-O-C (Leinmuller et al., 1991)5. 2. Tanin Hidrolisis Tanin hidrolisis merupakan ester dari glukosa dengan asam galat. Tanin ini dapat dihidrolisis dengan asam mineral panas menjadi gula dan asam-asam yang menjadi unsur pokoknya (Cheeke dan Shull, 1985). Menurut Hangerman (1992) 6, Interaksi tanin dipengaruhi oleh karakteristik protein seperti komposisi asam amino dan titik isoleotik serta karakteristik tanin seperti bobot molekul, temperatur, komposisi-komposisi pelarut dan waktu.
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Gambar 1. Struktur tanin terhidrolisis (a) dan terkondensasi (b) Sumber: (Dennis et al., 2005) 7
Menurut Susanti (2000)8, sifat utama tanin pada tanaman tergantung pada gugus fenolik-OH yang terkandung dalam tanin. Secara garis besar sifat tanin dapat dijabarkan sebagai berikut : 1. Tanin secara umum memiliki gugus fenol dan bersifat koloid. 2. Semua jenis tanin dapat larut dalam air, kelarutannya besar dan akan bertambah besar apabila dilarutkan dalam air panas. Begitu pula dalam pelarut organik seperti metanol, etanol, aseton dan pelarut organik lainnya. 3. Reaksi warna terjadi bila disatukan dengan garam besi. Reaksi ini digunakan untuk menguji klasifikasi tanin. Reaksi tanin dengan garam besi akan memberikan warna hijau dan biru kehitaman, tetapi uji ini kurang baik karena selain tanin yang dapat memberikan reaksi warna, zat-zat lain juga dapat memberikan reaksi warna yang sama. 4. Tanin mulai terurai pada suhu 98,8 0
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6. Ikatan kimia yang terjadi antara tanin-protein atau polimer lainnya terdiri dari ikatan hidrogen, ikatan ionik, dan ikatan kovalen. 7. Tanin mempunyai berat molekul tinggi dan cenderung mudah dioksidasi menjadi suatu polimer, sebagian besar tanin amorf (tidak berbentuk) dan tidak mempunyai mempunyai titik leleh. 8. Warna tanin akan menjadi gelap apabila terkena cahaya atau dibiarkan di udara terbuka. 9. Tanin mempunyai sifat bakteristatik dan fungistatik. Maserasi istilah aslinya adalah macerare (bahasa Latin, artinya merendam) : adalah sediaan cair yang dibuat dengan cara mengekstraksi bahan nabati yaitu direndam menggunakan pelarut bukan air (pelarut nonpolar) atau setengah air, misalnya etanol encer, selama periode waktu tertentu sesuai dengan aturan dalam buku resmi kefarmasian. Salah satu metode yang digunakan untuk uji kuantitatif senyawa tanin dalam teh adalah metode Lowenthal-procter ( Sudarmadji, 1984 ) 9. Metode ini menggunakan metode titrasi oksidasi yaitu dengan menggunakan senyawa pengoksidasi kalium permanganat. Penentuan kadar tanin dalam metode ini adalah berdasarkan jumlah gugus pada senyawa tanin. Titrasi dengan larutan kalium permanganat, gugus fenol pada tanin akan teroksidasi. Jumlah gugus fenol berbanding lurus dengan jumlah kalium permanganat yang diperlukan untuk titrasi. Pereaksi kalium permanganat bukan pereaksi baku primer. Sangat sukar untuk mendapatkan pereaksi ini dalam keadaan murni dan bebas dari mangan dioksida (Bassett, J,1994) 10.
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V.
ALAT DAN BAHAN
a. Alat-alat yang digunakan : 1. Erlenmeyer 100 mL
9. Buret makro 10. Batang pengaduk
2. Piala gelas
11. Botol semprot
3. Gelas ukur 50 mL
12. Statif dan klem
4. Pipet volumetrik
13. Neraca 13. Neraca analitik
5 dan 10 mL
14. Kertas Saring
5. Pipet tetes
15. Bunsen
6. Corong
16. Kaki tiga
7. Alas Titar 8. Neraca Analitik b. Bahan-bahan yang digunakan:
1. Sampel daun teh
6. Gelatin (C (C76H124O29 N24
2. Kalium Permanganat (KMnO4)
7. Garam jenuh (NaCl)
0,1N 3. Indigo Carmine (C16H8 N2 Na2O8S2
8. Asam sulfat (H2SO4
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VI.
CARA KERJA
1. Standarisasi KMnO4 Ditimbang Asam Oksalat 630 mgram
Dimasukan ke Labu Takar 100mL, ditera dan dihomogenkan dengan aquadest
Dipipet sebanyak 25mL Ke Erlenmeyer 250mL, ditambah H 2SO4 4N 25mL, dipanaskan hingga 700C
Dalam keadaan panas dititar dengan KMNnO 4, sampai timbul warna merah muda dari larutan tidak berwana
2. Penetapan Kadar tannin dalam sampel Sampel ditimbang 2,0 gram, ke gelas piala 100mL, ditambahkan air mendidih 20mL dan diamkan selama 20 menit
Titar dengan KMnO 0,1N
Dimasukan ke Labu takar 100mL ditera dan dihomogenkan dengan aquadest
Pipet 5mL filtrate,
Larutan tersebut disaring ke gelas piala kering
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3. Pembuatan Larutan Indigo Carmine
Ditimbang teliti 6 g indigo carmine
Ditambahkan asam sulfat 25 mL
Lalu ditambahkan air bebas air CO2 sampai menjadi 1 L
4. Pembuatan Larutan NaCl Asam Di pipet 975 mL larutan jenuh ke piala gelas
Ditambahkan 25 mL asam sulfat ekat
5. Pembuatan Gelatin p Ditimbang teliti 25 g gelatin
VII.
Ditambahkan 1 L NaCl jenuh
DATA PENGAMATAN
Nama Praktikan
:
1. Abdurrahman Risyad Baihaqi 2. Ni Luh Gede Mega Atikadevi
Dipanaskan hingga larut
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Deskripsi Sampel Uji : Daun kering, berwana coklat kehijauan, berbau khas khas teh
A. Kualitatif Identifikasi Bahan No. 1.
Nama Bahan
Rumus Molekul
Kalium
KMnO4
Permanganat
Sifat
o
Bentuk padatan
o
Tidak berbau
o
Larut dalam air berwana ungu
2.
Natrium
NaCl
o
putih
Klorida
3.
Gelatin
Bentuk padatan kristal
Gelatin
o
Larut dalam air
o
Tidak berbau
o
Gelatin berupa padatan
o
Tidak berbau
o
Tidak berasa Rapuh dalam keadaan
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o
Tidak berbau
o
Kaolin larut dalam air dingin atau air panas
5.
Asam Oksalat
H2C2O4.2H20
o
Asam oksalat merupakan senyawa kimia yang berbentuk kristal putih
6.
Asam Sulfat
H2SO4
o
Tidak larut dalam air
o
Wujud: cairan,
o
Warna : tidak bewarna,
o
Bau: tidak berbau.
o
Asam sulfat larut dalam air.
7.
Indigo Carmine
C16H8 N2 Na2O8S2
o
Pewarna makanan dan minuman, yang mengubahnya menjadi warna biru.
o
Berwujud padatan biru keunguan yang tidak
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B. Kuantitatif 1. Standarisasi KMnO4 0,1N Bobot zat
Volume
baku
KMnO4
primer (g)
(mL)
0,6298
23,47
23,50
= × ×
Baku primer
H2C2O4.2H2O H2C2O4.2H2O
Volume rata-rata :
=
Perhitungan [KMnO 4]
+ ,7 +,
=
629,8 23,50 ×63 ⁄ × 4 ( )
= 23,48 mL = 0,1064
⁄
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VIII. PERHITUNGAN
1. Standarisasi KMnO4 0,1 N a. Perhitungan bobot Asam Oksalat (H2C2O4) yang ditimbang
=25 ×63 ⁄ ×0,1 ⁄ × 4 (⁄) = 630 630 = 0,63 0,6300 b. Normalitas KMnO4 0,1 N
() ( ) = ) () × ( ( ) × ( 629,8 = 23,48 ×63 ⁄ × 4 ( ) = 0,1064
⁄
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1. Kadar Tannin dalam Sampel Teh
Tanin (%)
(A−B) A−B) ( ) = ()
x 100 %
( , −, ) , / = x 100% , = 9,38 % (
Simbol % tannin A
B
⁄)
Uraian Kadar senyawa tannin dalam sampel Volume KMnO4 yang bereaksi dengan total senyawa polifenol dalam sampel Volume KMnO4 yang bereaksi senyawa polifenol selain dari tanni dalam sampel
N Konsentrasi KMnO4 KMnO4
Satuan % (b/b)
mL
mL
mgrek/mL
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IX.
PEMBAHASAN
Percobaan kali ini adalah penetapan kadar tanin dalam teh dengan teknik maserasi. Tenik maserasi adalah bagian dari teknik ekstraksi. Ekstraksi adalah proses penarikan suatu zat dengan pelarut sehingga terpisah dari bahan yang tidak dapat larut dengan pelarut cair. Teknik ekstraksi sangat berguna untuk pemisahan secara cepat dan bersih, baik untuk zat organik atau anorganik, untuk analisis makro maupun mikro. Selain untuk kepentingan analisis kimia, ekstraksi juga banyak digunakan untuk pekerjaan preparatif dalam bidang kimia organik, biokimia, dan anorganik di laboratorium. Tujuan ekstraksi ialah memisahkan suatu komponen dari campurannya dengan menggunakan pelarut. Teknik maserasi adalah proses pengekstrakan simplisia dengan menggunakan pelarut dengan beberapa kali pengocokan atau pengadukan pada temperatur ruangan. Maserasi merupakan teknik yang bertujuan untuk menarik zat-zat berkhasiat yang tahan pemanasan maupun yang tidak tahan pemanasan. Pengikatan atau pelarutan zat aktif berdasarkan sifat kelarutannya dalam suatu pelarut (like dissolved like), penyarian zat
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pengerjaan dan peralatan yang sederhana. Namun metode ini juga memiliki kekurangan, yaitu cara pengerjaannya yang lama dan ekstraksi yang kurang sempurna. Tanin adalah suatu senyawa Polifenol dari tumbuhan,berasa Pahit dan Kelat yang bereaksi bere aksi dan dengan denga n menggumpal menggu mpalkan kan Protein Prot ein ,atau ,ata u senyawa sen yawa organik organ ik lainnya lain nya termasuk term asuk Asam Amino dan Alkohol. Tanin mempunyai kemampuan mengendapkan protein, karena tanin mengandung sejumlah kelompok ikatan fungsional yang kuat dengan molekul protein yang selanjutnya akan menghasilkan ikatan silang yang besar dan komplek yaitu protein tanin. Tanin mempunyai berat molekul 0,5-3 KD. Tanin alami larut dalam air dan memberikan warna pada air, warna larutan tanin bervariasi dari warna terang sampai warna merah gelap atau coklat, karena setiap tanin memiliki warna yang khas tergantung sumbernya. Penentuan
kadar
tanin
dalam
sampel
daun
teh
ini
melibatkan
titrasi
permanganatometri. Dalam praktikum ini, standarisasi dilakukan dengan penimbangan asam oksalat seberat 0,6298 gram, kemudian ditambahkan asam kuat yaitu H2SO4, dipanaskan
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antara KMnO4 dengan larutan baku tertentu. Hasil titik akhir titrasi pada saat proses standarisasi kalium permanganate ditunjukkan dengan adanya perubahan warna larutan menjadi merah muda. Kalium permanganat merupakan zat pengoksidasi yang sangat kuat. Pereaksi ini dapat dipakai tanpa tanpa penambahan indicator, indicator, karena mampu bertindak sebagai indikator. Permanganat dengan asam oksalat, dengan adanya asam sulfat, menghasilkan gas karbon dioksida: Reaksi ini lambat pada suhu kamar, tetapi menjadi cepat pada 70°C. maka diperlukan pemanasan sebelum dititrasi, dalam praktikum ini praktikan memanaskan larutan terlebih dahulu. Ion mangan(II) mengkatalisis reaksi ini; jadi, reaksi ini adalah otokatalitik; sekali ion mangan(II) telah terbentuk, reaksi menjadi semakin cepat. Pada proses titrasi permanganometri tidak perlu ditambahkan indikator untuk mengatahui terjadinya titik
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merupakan bahan baku primer yang baik; sangat murni, stabil selama pengeringan, dan tidak higroskopis. Titrasi oksalat oleh KMnO4 berlangsung pada larutan yang sudah dipanaskan sampai sekitar 70 °C dengan penambahan KMnO4 tidak terlalu cepat dan juga tidak terlalu lambat. Pemberian yang terlalu cepat cenderung menyebabkan reaksi antara MnO4- dengan Mn2+ (kesalahan positif), sedangkan jika terlalu l ambat mungkin terjadi kehilangan oksalat karena membentuk peroksida yang kemudian terurai menjadi air (kesalahan negatif). Pada standardisasi ini diperoleh konsentrasi KMnO4 sebesar 0.1064 N. Menetapkan
kadar
tanin
dalam
cara titrasi permanganat (metode Lowenthal). Lowenthal).
sampel
tehyaitu
dengan Metode
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kuantitatif dengan menggunakan menggunakan pipet volumetri dan dilakukan 2 kali atau secara duplo. Dilakukan penambahan 75 mL air suling dan indikator indigo carmine sebanyak 5 mL. Kemudian dititrasi dengan KMnO4 yang telah distandarisasi terlebih dahulu. Perubahan warna yang terjadi dalam titrasi adalah dari warna yang semula biru saat ditambahkan indigo carmine, setelah dititrasi menjadi warna kuning. Fungsi indigo carmine adalah sebagai indikator perubahan warna dalam titrasi ini. Volume pentitar yang dikeluarkan adalah volume A atau volume total polifenol dalam sampel. Pada pemipetan sampel 10 mL, dilakukan secara duplo dengan penambahan 10 mL NacL asam, 2 mL gelatin , dan 2 gram kaolin. Penambahan NaCl asam adalah untuk mengendapkan senyawa tannin yang terkandung di dalam sampel teh, sedangkan fungsi dari gelatin dan kaolin adalah
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Kadar tanin yang diperoleh diperol eh dari pengujian kadar tanin dalam daun teh adala h 9,38 %. Kadar tanin yang diperoleh ini Sesuai dengan literatur yaitu kadar dalam teh biasanya 9-20%. kadar tanin yang diukur dengan kadar tanin sebenarnya mungkin terutama disebabkan oleh kurang optimalnya proses isolasi yang dilakukan seperti kurang lamanya proses pros es maseras mas erasii yang dilakuka dil akukan n sehingga sehi ngga tidak tida k seluruh selu ruh tanin tani n terekst ter ekstrak rak dar i sampel sam pel kasarnya.
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XI.
DAFTAR PUSTAKA
a. Reaski
1. Bassett j, R.CDenny, G.H Jeffery, J. Mendham. 1994. Buku 1994. Buku Ajar Vogel Kimia Analisis Kuantitatif anorganik. Edisi 4. Penerjemah : A. Hayana Pudjaatmaka. Jakarta: Penerbit buku Kedokteran EGC. (halaman 409) 2. Harbone J B.1987. Metode B.1987. Metode fitokimia penuntun cara modern menganalis tumbuhan. Terbitan kedua. Terjemahan oleh pandmawnata kosasih. Press. Bandung. (halaman 107) b. Landasan Teori 1. Winarsi, Hery. 2007. Antioksidan 2007. Antioksidan alami dan radikal radikal bebas. Kanisius:
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7. Dennis, O., W. J. M. Smith., J. D. Brooker, & M. C. ScWeeney. 2005
Tolerance mechanisms of streptococci to hydrolysable and condensed tannins. tannins . Anim Feed Sci. Technol. 121: 59-75 8. Susanti, C. M. E. 2000. Autokondensat tanin sebagi sebagi perekat kayu lamina. lamina.
Jurusan IPK. Program pasca sarjana IPB. Bogor. Desertasi
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LAMPIRAN I.
IDENTIFIKASI BAHAN KIMIA
Gambar
Rumus molekul Rumus bangun
Berat
Nama
molekul
senyawa
Sifat fisika Sifat kimia
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Merupakan zat pengoksidasi yang kuat
Bereaksi
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H 2O
18 g/mol
Air
Cairan, tidak berwarna,tida k berbau
Titik 100◦C
didih
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elektronegatif nya lebih dari 2
Larutannya merupakan
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berwarna
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II.
BAGAN KERJA
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