Isolasi dan identifikasi flavonoid infusa daun Srikaya (Annona squamosa, L) dan uji antiproliferasi terhadap sel HeLa

NURANI, Laela Hayu, Dr. Suwidjiyo Pramono ,Apt

2004 | Tesis | S2 Ilmu Farmasi

Telah dilakukan penelitian isolasi dan identifikasi flavonoid infus daun srikaya (Annona squamosa, L) dan uji efek antiproliferatifnya terhadap sel HeLa. Penyarian dilakukan dengan metode infundasi selama 30 menit kemudian diuapkan. Ekstrak ini kemudian dihidrolisis dengan HCl dan difraksi dengan eter. Flavonoid dalam fraksi eter diisolasi menggunakan kromatografi kertas preparatif dengan fase gerak asam asetat 15 % dan diperoleh 4 bercak di bawah sinar UV 366 nm dengan harga Rf berturut-turut dari bercak 1 sampai 4 adalah 0,11; 0,23; 0,46; dan 0,78. Isolat diperiksa kemurniannya dengan kromatografi kertas dua dimensi menggunakan fase gerak yang berbeda.Pemeriksaan flavonoid selanjutnya menggunakan pereaksi diagnostik dengan metode spektrofotometri ultraviolet. Pencatatan spektra dilakukan pada panjang gelombang 200-500 nm. Berdasarkan analisis data baik dari reaksi warna, kromatografi kertas maupun spektrofotometri ultraviolet menunjukkan bahwa flavonoid Annona squamosa, L isolat-1 adalah 5-hidroksi 7-R flavonol, isolat-2 adalah 5, 3`, 4` trihidroksi 7-R flavonol, isolat-3 adalah 5 hidroksi 7 R flavanon, isolat-4 adalah 7 R flavanon. Infusa dan isolat-isolat diuji aktivitas sitotoksisitasnya terhadap sel HeLa. Diperoleh harga LC50 terhadap sel HeLa, infus = 889,5; isolat-1 = 12,63; isolat-2 = 18,47; isolat-3 = 75,74; isolat-4 = 204,4 μg/ml. Kinetika pertumbuhan sel HeLa akibat bahan uji diketahui dari harga doubling time. Hasil doubling time menunjukkan bahwa isolat flavonoid mempunyai peran dalam menghambat proliferasi sel HeLa.

The flavonoid isolation and identification of srikaya (Anona squamosa, L) leaf infusion and the anti proliferative effect toward HeLa cell had been researched in this thesis. The extraction was conducted with the infundation method for 30 minutes followed by the evaporation process. The extract then hydrolated with Hcl and fracted with eter. Flavonoid in the ether fraction was isolated using preparative paper chromatography with asetil acid 15% as the mobile phase and 4 spots were obtain under the 366 nm UV rays with the Rf value subsequently 0.11; 0.23;0.46 and 0.78. The purification of isolat was examined with two dimension paper chromatography with the different mobile phase. The next flavonoid examination used diagnostic reactor with the ultraviolet spectroscopy method. The spectroscopy process was conducted at 200- 500 nm wave length. Based on the data analysis from color reaction, paper chromatography and ultraviolet spectroscopy, the flavonoid of Annona squamosa L were 5-hidroksi 7-R flavonol for isolat 1; 5,3’, 4’ trihidroksi 7-R flavonol for isolat-2; 5 hidroksi 7-R flavonon for isolat 3 and 7-R flavonon for isolat-4 The infusion and the isolates were tested its cytotoxcicity towards HeLa cell. The LC50 value to the HeLa cell were obtained, infus : 0.8895, isolat-1 = 0.01263, isolat-2 = 0.01847, isolat-3 = 0,07574 and isolat-4 = 0,2044 mg/ml The kinetics growth of HeLa cell as the result of the material tested were recognized from the doubling time. The result of doubling time showed that the flavonoid isolat has a role to obstruct the HeLa cell proliveration.

Kata Kunci : Obat Tradisional,Daun Srikaya,Uji Antiproliferasi