Isolasi dan Identifikasi Oligosakarida Madu Hutan Gunung Tambora Sumbawa

Abstract

Prebiotic activity of oligosaccharides isolated from the honey of Mount Tambora’s forests in Sumbawa has been studied. This activity were strongly believed occured because of the oligosaccharides with DP 3 - 4. However, the oligosaccharides compounds of Mount Tambora’s forests in Sumbawa have never been studied yet. The objectives of this study are to isolate and identify oligosaccharides with DP 3 and 4, and to obtain the optimum method of the process. The preparation of sample has been carried out through the activated charcoal separation by stirring process within 10% ethanol solution to remove the monosaccharides, and 50% ethanol solution to recover the oligosaccharides compounds. Detection and isolation process of oligosaccharides were carried out by using Thin Layer Chromatography (TLC) with buthanol: acetic acid: water (3:1:1 v/v) as an eluen solvent. Glucose (DP 1) and maltose (DP 2) were used as standards. Honey and isolated oligosaccharides samples were identified by using Liquid Chromatography-Mass Spectrometry (LC-MS) with glucose, maltotriose (DP 3), and maltotetraose (DP 4) as a standard. Preparation of samples resulted in 2.09 % yields. Isolated oligosaccharides resulted in 3 spots with retention factors of 0.38 (DP 1), 0.31 (DP 2), and 0.22 (DP>2) respectively. Identified oligosaccharides showed that carbohydrates with DP 1, DP 3, and DP 4 are present in honey and Isolated oligosaccharides samples, respectively 2182.96, 246.94, dan 118.71 (μg/mL) in honey, and 278.34, 95.03, and 118.71 (μg/mL) in isolated oligosaccharides samples.

Isolat oligosakarida madu hutan Gunung Tambora Sumbawa telah terbukti memiliki aktivitas prebiotik. Aktivitas prebiotik tersebut diyakini berasal dari oligosakarida dengan DP 3-4. Namun, kandungan oligosakarida madu hutan Gunung Tambora Sumbawa belum pernah diteliti sebelumnya. Tujuan penelitian ini adalah mengisolasi dan mengidentifikasi oligosakarida madu dengan DP 3-4 serta mendapatkan optimasi metode dalam prosesnya. Preparasi sampel dilakukan dengan adsorpsi arang aktif dan pengadukan dalam etanol 10% untuk menghilangkan monosakarida serta etanol 50% untuk pemulihan oligosakarida. Deteksi dan isolasi oligosakarida dilakukan dengan Kromatografi Lapis Tipis (KLT) menggunakan campuran eluen butanol: asam asetat: air (3:1:1 v/v) dengan glukosa (DP 1) serta maltosa (DP 2) sebagai standar. Identifikasi dilakukan menggunakan Liquid Chromatography-Mass Spectrometry (LC-MS) dengan glukosa, maltotriosa (DP 3), dan maltotetraosa (DP 4) sebagai standar. Preparasi oligoskarida menghasilkan 2.09% rendemen. Isolasi oligosakarida menghasilkan 3 spot dengan Rf berturut-turut: 0.38 (DP 1), 0.31 (DP 2), dan 0.22 (DP>2). Hasil identifikasi menunjukkan bahwa madu murni mengandung karbohidrat DP 1, DP 3, dan DP 4 berturut-turut 2182.96, 246.94, dan 118.71 (μg/mL) sedangkan isolat oligosakarida mengandung karbohidrat DP 1, DP 3, dan DP 4 berturut-turut 278.34, 95.03, dan 118.71 (μg/mL).