laporan resmi praktikum fitokimia

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LAPORAN RESMI PRAKTIKUM FITOKIMIA

PERCOBAAN II PEMISAHAN, IDENTIFIKASI FRAKSI KURKUMINOID DARI RIMPANG TEMULAWAK (Curcuma xanthorrhizae Rhizoma) SECARA KROMATOGRAFI KOLOM

Oleh : 1. UKI SEPTI 2. VERONICA LESLIE 3. VIVIN WIDYA N 4. VRIHATDIAN F 5. WENSILAUS B 6. WIDITA DILAH 15092789A 15092790A 15092791A 15092793A 15092794A 15092795A

DOSEN PENGAMPU : MAMIK PONCO RAHAYU,M.si,Apt.

TEORI III FAKULTAS FARMASI UNIVERSITAS SETIA BUDI SURAKARTA 2012 PERCOBAAN II

PEMISAHAN, IDENTIFIKASI FRAKSI KURKUMINOID DARI RIMPANG TEMULAWAK (Curcuma xanthorrhizae Rhizoma) SECARA KROMATOGRAFI KOLOM TUJUAN Setelah melakukan praktikum ini mahasiswa dapat memahami dan melakukan: Ekstraksi senyawa organik dari simplisia tanaman dengan metode refluk Pemisahan komponen-komponen senyawa kimia dengan kromatografi kolom vakum Identifikasi senyawa secara kromatografi lapis tipis

PENDAHULUAN Kunyit merupakan tanaman berbatang basah dan mempunyai tinggi sampai 1 meter. Tanaman ini dapat tumbuh di Kingdom Divisi Sub Divisi Kelas Ordo Famili Genus Spesies berbagai tempat. Kunyit(Curcuma Domestica Valet) termasuk dalam klasifikasi sebagai berikut: : Plantae (tumbuh-tumbuhan) : Spermatophyta (tumbuhan berbiji) : Angiospermae (berbiji tertutup) : Monocotyledonae (biji berkeping satu) : Zingiberales : Zingiberaceae : Curcuma : Curcuma xanthorrhizae Rhizoma

Susunan temulawak terdiri atas akar, rimpang, batang semu, pelepah daun, daun, tangkai bunga dan kuntum bunga. Komponen utama yang terpenting dalam rimpang temulawak adalah kurkuminoid dan minyak atsiri. Berdasarkan hasil penelitian Balai Penelitian Tanaman Rempah dan Obat (Balittro) bahwa kandungan kurkumin rimpang temulawak rata-rata 10,92 %. Selain temulawak, ada juga beberapa tanaman yang mengandung kurkumin, misalnya kunyit dan temu hitam. Sifat-sifat kurkumin adalah sebagai berikut : Berat molekul : 368,37 (C = 68,47 %; H = 5,47 %; O = 26,06 %)

Warna

: light yellow

Melting point : 183o C Larut dalam alkohol dan asam asetat glasial Tidak larut dalam air Kurkumin dapat larut dalam alkohol dan asam asetat glasial. Alkohol merupakan zat cair yang mudah menguap, tidak berwarna dan memiliki bau yang khas. Titik didih alkohol 118,1 C, spesific gravity 1,049, berat molekul 60,05 gram/mol. Salah satu cara pengambilan kurkumin dari rimpangnya adalah dengan cara ekstraksi. Ekstraksi merupakan salah satu metode pemisahan berdasarkan perbedaan kelarutan. Secara umum ekstraksi dapat didefmisikan sebagai proses pemisahan dan isolasi zat dari suatu zat dengan penambahan pelarut tertentu untuk mengeluarkan komponen campuran dari zat padat atau zat cair. Dalam hal ini fraksi padat yang diinginkan bersifat larut dalam pelarut (solvent), sedangkan fraksi padat lainnya tidak dapat larut. Proses tersebut akan menjadi sempurna jika solute dipisahkan dari pelarutnya, misalnya dengan cara distilasi/penguapan. Kromatografi lapis tipis menggunakan sebuah lapis tipis silika atau alumina seragam pada sebuah lempeng gelas atau logam atau plastik yang keras. Jel silika (atau alumina) - fase diam. Fase diam untuk kromatografi lapis tipis seringkali juga mengandung substansi yang mana dapat berpendarflour dalam sinar ultra violet, alasannya akan dibahas selanjutnya. Fase gerak merupakan pelarut atau campuran pelarut yang sesuai. Nilai Rf adalah karakteristik suatu komponen sehingga dapat membedakan satu dengan lainnya. 010009000003a85203000000aff1010000001610000026060f002220574d464301000 00000000100f3f4000000001600000000200000c09d0200c0bd0200010000006c0000 0000000000000000001b0100002a0100000000000000000000242700003629000020 454d4600000100c0bd02000c00000001000000000000000000000000000000560500

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Gambar 2. Struktur kurkuminoid dari rimpang temulawak

ALAT DAN BAHAN 1. Ekstraksi: Alat : labu alas bulat, kondensor, selang air, klem dan statif, kaki tiga, lampu spirtus, panci, kertas saring, corong, kapas, erlenmeyer. Bahan : serbuk temulawak, petroleum eter, metanol. 2. KKV Alat : kolom, vakum, beaker glass, batang pengaduk, botol. Bahan : ekstrak temulawak, heksan, etil asetat, metanol. 3. KLT

Alat : bak kromatografi, lempeng KLT, silika gel GF254, beaker glass, pipa kapiler, kertas saring, papan kromatografi. Bahan : ekstrak temulawak, fraksi, metnol, heksan, etil asetat, asam asetat. CARA KERJA A. Penyiapan Ekstrak Ekstraksi serbuk rimpang temulawak (50 g) secara refluk dengan 100 ml pelarut petroleum eter selama 1 jam. Setelah itu disaring dan ampas dikeringkan. Ekstraksi ampas secara refluk dengan methanol sebanyak 100 ml selama 2 jam. Saring filtrat kemudian uapkan di atas tangas air pada suhu serendah mungkin hingga bebas pelarut. Kemudian ambil sedikit dalam wadah vial kecil dan gunakan untuk pemeriksaan KLT dengan tujuan mencari eluen yang terbaik untuk pemisahan selanjutnya. Skema Padat-cair

Ekstraksi cair-cair

B. Pemisahan komponen senyawa aktif dengan KKV Masukkan adsorben ke dalam kolom dalam keadaan kering atau dibuat seperti bubur dengan pelarut. Pengisian dilakukan dengan bantuan pemampat (pengaduk) untuk memampatkan adsorben dan glass wool pada dasar kolom. Pengisian harus dilakukan secara hati-hati dan sepadat mungkin agar rata

sehingga terhindar dari gelembung-gelembung uadara. Masukkan ekstrak yang sudah dikeringkan dengan adsorben dengan hati-hati ke atas kolom kemudian elusi dengan eluen n-hexan, etil asetat, methanol secara gradien. Dengan bantuan vakum, masing-masing eluat yang keluar ditampung dalam wadah, kemudian uapkan. Lalu identifikasi fraksi secara KLT dengan eluen yang cocok sehingga diperoleh pemisahan yang baik. Fraksi yang mempunyai Rf sama dengan standar (kurkumin, kurkuminoid) dikumpulkan. Pemisahan senyawa kurkumin dapat dilanjutkan dengan kromatografi kolom atau secara kromatografi preparatif.

Kromatografi Lapis Tipis

Kromatografi Lapis Tipis Preparatif

Sublimasi

HASIL PENGAMATAN Berat kurkuminoid Serbuk+plastik Plastik Beratkurkuminoid : 51,268 gram : 1,090 gram : 50,178 gram

Setelah di refluk Serbuk + kertas folio : 55,597 gram Kertas folio Beratserbuk : 7,733 gram : 47,864 gram

Ekstrak kental Beaker + ekstrakkental Beaker Ekstrakkental

: 101,358 gram : 99,821gram : 1,537gram

Rendemen ekstrak n-heksan = = Eluen yang digunakan 1. N-Heksan = 100 ml

2. N-Heksan : etilasetat = 8 : 2 (40 ml : 10 ml) 3. N-Heksan : etilasetat = 6 : 4 (30 ml : 20 ml) 4. N-Heksan : etilasetat = 4 : 6 (20 ml : 30 ml) 5. N-Heksan: etilasetat = 2 : 8 (10 ml : 40 ml) 6. Etilasetat = 50 ml

7. Etilasetat : methanol = 8 : 2 (40 ml : 10 ml) 8. Etilasetat : methanol = 6 : 4 (30 ml : 20 ml) 9. Etilasetat : methanol = 4 : 6 (20 ml : 30 ml) 10. Etilasetat : methanol = 10 : 40 (10 ml : 40 ml) 11. Methanol = 50 ml

HASIL IDENTIFIKASI SECARA KLT

Gambar : UV 254

Keterangan : Totolan : 1. n-hexan 2. Standart kurkumin 3. n-hexan 100% 4. n-hexan : etil asetat 8:2 5. n-hexan : etil asetat 6:4 6. n-hexan : etil asetat 4:6 7. n-hexan : etil asetat 2:8 8. etil asetat 100% 9. etil asetat : methanol 8:2

10. etil asetat : methanol 6:4 11. etil asetat : methanol 4:6 12. etil asetat : methanol 2:8 13. methanol 100%

Gambar : UV 366

Keterangan : Totolan : 1. n-hexan 2. Standart kurkumin 3. n-hexan 100% 4. n-hexan : etil asetat 8:2 5. n-hexan : etil asetat 6:4 6. n-hexan : etil asetat 4:6 7. n-hexan : etil asetat 2:8 8. etil asetat 100% 9. etil asetat : methanol 8:2 10. etil asetat : methanol 6:4 11. etil asetat : methanol 4:6 12. etil asetat : methanol 2:8 13. methanol 100% Gambar : Cahaya Tampak

Keterangan : Totolan : 1. n-hexan 2. Standart kurkumin 3. n-hexan 100% 4. n-hexan : etil asetat 8:2 5. n-hexan : etil asetat 6:4 6. n-hexan : etil asetat 4:6 7. n-hexan : etil asetat 2:8 8. etil asetat 100% 9. etil asetat : methanol 8:2 10. etil asetat : methanol 6:4 11. etil asetat : methanol 4:6 12. etil asetat : methanol 2:8 13. methanol 100% Hasil KromatogramNo. Bercak 1. 2. 3. 4. 5. 6. 7. n-heksan Standartkurkumin N-heksan 100% n-heksan:etiasetat 8:2 n-heksan:etilasetat 6:4 n-heksan:etilasetat 4:6 n-heksan:etilasetat 1,78 Sinar Tampak Kuning Kuningkecoklat Kuning UV254 Ungu Hijau Ungu Ungu UV366 Kuning Kuning coklat Kuning Fraksi Rf Warna Bercak dengan Penampak Noda

8. 9. 10. 11. 12. 13.

2:8 Etilasetat Etilasetat:methanol 8:2 Etilasetat:methanol 6:4 Etilasetat:methanol 4:6 Etilasetat:methanol 2:8 Methanol 100%

1,86 1,82 -

Kuningkecoklat Kuningkecoklat Coklat Coklat Coklat

Hijau Hijau Ungu Hijau Hijau Hijau

Kuningkecoklat Kuningkecoklat Kuning Coklat Coklat

Hasil Fraksi PenggabunganNo. 1 2 3 Fraksipenyusun 5,6 8,9 10-13 Berat

PEMBAHASAN Ekstraksi adalah penyarian zat-zat aktif dari bagian tanaman obat. Adapun tujuan dari ekstraksi yaitu untuk menarik komponen kimia yang terdapat dalam simplisia, suatu proses pemisahan dari bahan padat maupun cair dengan bantuan pelarut. Pelarut yang digunakan berfungsi mengekstrak substansi yang diinginkan tanpa melarutkan material lainnya (Anonim 2008). Faktor-faktor yang mempengaruhi laju reaksi adalah tipe persiapan sampel, waktu ekstraksi, kuantitas pelarut, suhu pelarut, dan tipe pelarut. Semakin lama waktu ekstrak dan semakin halus ekstraknya maka semakin banyak pula rendemen yang didapatkan. Pemilihan methanol sebagai pelarut karena sifatnya yang mampu melarutkan hampir semua zat, baik yang bersifat polar, semipolar, dan non polar serta kemampuannya untuk mengendapkan protein, menghambat kerja enzim sehingga dapat terhindar proses hidrolisis dan oksidasi, dan larutan ini bersifat polar sehingga mempunyai titik didih yang tinggi sehingga memiliki kelarutan yang tinggi dan dapat larut sempurna. Methanol yang digunakan adalah 100% yang biasanya digunakan untuk ekstraksi sample segar (bahan hayati). Pelarut methanol 100% lebih cepat

mengekstrak temulawak karena methanol 100% memiliki titik didih (suhu) yang lebih tinggi sehingga lebih mudah terekstrak. Ada beberapa metode yang digunakan untuk mengekstrak komponen dari temulawak yaitu metode soxhlet, penggodokan (refluks).Sokletasi merupakan metode yang paling simple. Waktu maksimum sokletasi adalah 8 jam. Keuntungan dari metode ini adalah dapat digunakan sampel dengan tekstur yang lunak dan tidak tahan terhadap pemanasan secara langsung, digunakan pelarut yang lebih sedikit, dan pemanasannya dapat diatur. Kerugian dari metode ini karena pelarut didaur ulang sehingga ekstrak yang terkumpul di sebelah bawah wadah terus-menerus dipanaskan sehingga dapat menyebabkan reaksi peruraian oleh panas adalah methanol 100%. Metode refluks mempakan metode dimana sampel yang dicampur dengan pelarutnya sehingga tidak bisa dihasilkan ekstrak yang murni. Pelarut yang digunakan adalah petroleum eter. Kesalahan-kesalahan yang terjadi dari proses ekstraksi kali ini adalah waktu pemekatan fraksi hasil terlalu kering,dan sisa fraksi yang dipekatkan terbuang sebelum melakukan penimbangan,sehingga rendemen tidak dapat dihitung. Solusi agar kesalahan ini tidak terjadi adalah dengan memperhatikan prinsip kerja yang baik, teliti dalam pengukuran volume cairan pelarut, dan menggunakan pelarut yang sesuai. GAMBAR ALAT dan terbatas dengan pelarut murni atau campuran azeotropik (Anonim 2008). Pelarut yang digunakan

Gambar : Sokhlet

Gambar : Refluks

KESIMPULAN Ekstraksi adalah proses pengambilan senyawa tunggal atau majemuk dari suatu bahan dengan menggunakan pelarut tertentu berdasarkan distribusinya pada dua fase yang tidak saling bercampur. Bahan hayati yang digunakan adalah temulawak Metode ekstraksi yang digunakan meliputi metode soxhiet, penggodokan (refluks). Faktor-faktor yang mempengaruhi laju reaksi adalah tipe persiapan sampel, waktu ekstraksi, kuantitas pelarut, suhu pelarut, dan tipe pelarut. Dari praktikum kali ini hasil Rf yang mempunyai nilai Rf sama dengan standar kukurmin adalah etil asetat 100% dan etil asetat : methanol = 8:2.

DAFTAR PUSTAKA 1. Anonim. 2008. Ekstraksi. [terhubung berkala].

medicafarma.blogspot.com12008/111ekstraksi.html. [13 November 2009]

2. Anonim.

2008. Temulawak

(Curcuma xanthoma,

Roxb).

[terhubung

berkala]. http://www.pusri.co.id/budidaya/obat/temulawak.pd [13 November 2009]

3. Anonim, 1997 , Materi Medika Indonesia, Jilid I,II,III.IV, Departemen Kesehatan RI,Jakarata 4. Anonim , 1986,Sediaan Galenik, 25 26,Depkes RI, Jakarta. 5. Titik Sunarni,Mamik Ponco Rahayu,Fransiska Leviana, 2011, Buku Petunjuk Praktikum Fitokimia S1 Farmasi, Universitas Setia Budi, Surakarta.