FRAKSI ANTIPLASMODIUM PALING AKTIF DARI DAUN KAPUR (Harmsiopanax aculeatus Harms) DAN IDENTIFIKASI BEBERAPA KANDUNGAN SENYAWANYA MENGGUNAKAN GC-MS

Rachel Turalely, S.Pd, Dr. Ratna Asmah Susidarti, M.Si., Apt.

2011 | Tesis | S2 Bioteknologi

Malaria merupakan penyakit menular yang disebabkan oleh Plasmodium yang masih menjadi masalah kesehatan dunia, terutama di negara-negara yang sedang berkembang. Adanya resistensi Plasmodium terhadap klorokuin sebagai obat pilihan pertama untuk malaria telah mendorong dilakukannya penelitian untuk menemukan dan mengembangkan obat baru terutama yang berasal dari bahan alam. Daun tanaman kapur (Harmsiopanax aculeatus Harms) telah digunakan secara tradisional oleh masyarakat Maluku untuk mengobati malaria. Meskipun demikian belum ada penelitian fitokimia maupun farmakologi yang dilakukan terhadap tanaman ini. Penelitian ini bertujuan untuk mendapatkan fraksi paling aktif antiplasmodium daun kapur dan menganalisis kandungan kimianya. Serbuk kering daun kapur sebanyak 1,3 kg diekstraksi secara maserasi bertingkat menggunakan n-heksan, etil asetat dan metanol. Setelah pelarut diuapkan diperoleh ekstrak heksan sebesar 15,6 g (1,2%), ekstrak etil asetat 53,3 (4,1%) dan ekstrak metanol 61,1 (4,7%). Ekstrak tersebut diuji aktivitas antiplasmodiumnya secara in vivo menggunakan metode 4 days suppressive test pada mencit Swiss yang diinfeksi Plasmodium berghei. Persentase parasitemia dihitung menggunakan apusan darah tipis yang dicat Giemsa dan diperiksa di bawah mikroskop cahaya. ED50 ekstrak heksan, etil asetat dan metanol berturut turut adalah 467,58; 2074,02 dan 16,16 mg/kgBB. Fraksinasi ekstrak metanol dilakukan dengan VLC menggunakan silika gel yang dielusi dengan meningkatkan polaritas pelarut, menghasilkan 18 fraksi gabungan (FG1 – FG8). FG tersebut diuji Hem Polymerization Inhibitory Activity (HPIA) secara in vitro. FG8 adalah fraksi paling aktif dengan IC50 HPIA 18,22 µg/ml. Uji fitokimia terhadap fraksi ini menggunakan pereaksi semprot memperlihatkan keberadaan minyak atsiri, triterpenoid dan senyawa fenolik. Pemisahan FG8 menggunakan kromatografi tekan menghasilkan 19 fraksi gabungan (FG8.1 – FG8.19). Fraksi yang mengandung spot berfluoresensi biru (FG8.5) dipisahkan menggunakan PLC 4 kali elusi dengan kloroform. Pita berfluoresensi biru dikerok dan diekstraksi dengan kloroform. Isolat kemudian dianalisis menggunakan GC-MS. Tujuh komponen utama dengan komposisi persentase lebih dari 3,11% diidentifikasi sebagai eugenol (tr = 12,692; 18,22%), isopropil miristat (tr = 16,333; 3,99%); bis(2- metilpropil) ftalat (tr = 16,942; 7,15%); metil palmitat (tr = 17,442; 3,11%); asam palmitat (tr = 17,883; 25,72%); butil 2-metilpropil ftalat (tr = 17.957; 9.37%) dan bis(2-etilheksil) ftalat (tr = 23,258; 20,29%) berdasarkan spektra massa senyawa tersebut yang dibandingkan dengan data spektra massa WILLEY229 Library.

Malaria is an infectious disease caused by Plasmodium which is still a worldwide health problem, especially in the developing countries. The rising of Plasmodium resistance towards chloroquine as a first line drug for malaria have encouraged to discover and develop new drugs mainly derived from natural sources. Harmsiopanax aculeatus (kapur plant) has traditionally used by people of in Maluku Province to treat malaria. However, there was no phytochemical and pharmacological studies have been done on this plant. The aims of this study were to obtain the most active antiplasmodial fraction from kapur leaves and to identify its chemical constituents. The dried powder of Kapur leaves (1.3 kg) were extracted successively by maceration with n-hexane, ethyl acetate and methanol. After removal the solvents the hexane 15.6 g (1.2%), ethyl acetate 53.3 g (4.1%) and methanol 61.1 g (4.7%) extracts were obtained. Those extracts were assayed for their in vivo antiplasmodial activities by using 4-days suppressive test in Swiss mice infected with Plasmodium berghei. The percentage of parasitemia was calculated using thin blood smears which were stained by Giemsa and observed under light microscope. The ED50 of hexane, ethyl acetate and methanol extracts were 467.58, 2074.02 and 16.16 mg/kgBW, respectively. Fractionation of the methanol extract by vacuum liquid chromatography over silica gel eluted by solvents of increasing polarity gave 18 combined fractions (FG1 – FG18). Those fractions were tested for their in vitro Hem Polymerization Inhibitory Activity (HPIA). FG8 was the most active fraction with the IC50 HPIA of 18.22 µg/ml. Phytochemical test of this fraction using spray reagent showed the existence of essential oils, triterpenoids, and phenolic compounds. Separation of FG8 using pressed chromatography gave 19 combined fractions (FG8.1-FG8.19). The fraction containing intense blue fluorescent spot (FG8.5) was further separated by PLC fourthly eluted with chloroform. The blue fluorescent band was scrapped and then extracted with chloroform. The isolate was then analyzed by GC-MS. Seven major components with the percentage of compotition more than 3.11% were identified as eugenol (tr = 12.692; 18.22%), isoprophyl myristate (tr = 16.333; 3.99%); bis(2- methylpropyl) phtalate (tr = 16.939; 7.15%); methyl palmitic (tr = 17.442; 3.11%); palmitic acid (tr = 17.883; 25.72%); butyl 2-methylpropyl phtalate (tr = 17.957; 9.37%) and bis(2-ethylhexyl) phtalate (tr = 23.258; 23%) based on their mass spectra compared with the WILLEY229 library of mass spectral data.

Kata Kunci : Harmsiopanax aculeatus Harms, Plasmodium berghei, Hem Polymerization Inhibitory Activity.