LAPORAN RESMI PRAKTIKUM FITOKIMIA

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

Oleh :
1. UKI SEPTI

15092789A

2. VERONICA LESLIE

15092790A

3. VIVIN WIDYA N

15092791A

4. VRIHATDIAN F

15092793A

5. WENSILAUS B

15092794A

6. WIDITA DILAH

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

berbagai

tempat. Kunyit(Curcuma

Domestica Valet) termasuk dalam klasifikasi sebagai berikut:
Kingdom

: Plantae (tumbuh-tumbuhan)

Divisi

: Spermatophyta (tumbuhan berbiji)

Sub Divisi

: Angiospermae (berbiji tertutup)

Kelas

: Monocotyledonae (biji berkeping satu)

Ordo

: Zingiberales

Famili

: Zingiberaceae

Genus

: Curcuma

Spesies

: 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.
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f7fff7fff7fff7fff7fff7fff7fff7fff7fff7fff7

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

: 51,268 gram

Plastik
Beratkurkuminoid

: 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 Kromatogram
No.

Fraksi

Rf

Warna Bercak dengan Penampak Noda

1.
2.
3.
4.
5.

n-heksan
Standartkurkumin
N-heksan 100%
n-heksan:etiasetat 8:2
n-heksan:etilasetat

1,78
-

Sinar Tampak
Kuning
Kuningkecoklat
-

6.

6:4
n-heksan:etilasetat

-

Kuning

Ungu

7.

4:6
n-heksan:etilasetat

-

-

-

Bercak
UV254
Ungu
Hijau
Ungu

UV366
Kuning
Kuning coklat
Kuning
-

8.
9.

2:8
Etilasetat
Etilasetat:methanol

1,86
1,82

Kuningkecoklat
Kuningkecoklat

Hijau
Hijau

Kuningkecoklat
Kuningkecoklat

10.

8:2
Etilasetat:methanol

-

-

Ungu

-

11.

6:4
Etilasetat:methanol

-

Coklat

Hijau

Kuning

12.

4:6
Etilasetat:methanol

-

Coklat

Hijau

Coklat

13.

2:8
Methanol 100%

-

Coklat

Hijau

Coklat

Hasil Fraksi Penggabungan
No.
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

dan terbatas dengan

pelarut murni atau campuran azeotropik (Anonim 2008). Pelarut yang digunakan
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

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.

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