Judul: Profil Bioaktif Ekstrak dan Fraksi Larut Lemak Sargassum cristaefolium dari Indonesia dan Evaluasi Aktivitas Anti-Inflamasi dan Antioksidan secara In-vitro
Abstrak: Sargassum is a popular brown seaweed genus which has been traditionally utilized by East Asian community as food and medicine. This brown seaweed is reported to exhibit several biological activities which promote human health, including anti-inflammatory and antioxidant activity. This research aimed to evaluate the potency of Indonesian S. cristaefolium as a source of antiinflammatory and antioxidant agents. To achieve the aforementioned general objective, this research was carried out through three main stages, i.e. 1) preliminary chemical characterization of brown seaweed obtained from four different water areas in Java - Indonesia, 2) preliminary screening of anti-inflammatory and antioxidant activity of brown seaweed crude lipid extracts, and 3) antiinflammatory and antioxidant evaluation of lipid-soluble fractions derived from selected brown seaweed sample and putative identification of bioactive compounds using ultra-high-performance liquid chromatography electrospray ionization orbitrap tandem mass spectrometry (UHPLC-ESI-ORBITRAP-MS/MS) instrument. Research was conducted at Chemistry Laboratory of Southeast Asian Food and Agricultural Science and Technology (SEAFAST) Center, Primate Research Center, and Tropical Biopharmaca Research Center of IPB University, Fat and Oil Laboratory, Food Science and Technology Faculty – University Putra Malaysia (UPM), and Institute of Bioscience of UPM. Results of preliminary chemical characterization of brown seaweeds obtained from different coastal areas showed that samples from southern waters of Java (Sayang Heulang Beach/SHB and Ujung Genteng Beach/UGB) were characterized by high contents of ash, alginate and fucose-containing sulfated polysaccharides (FCSPs) yield, and FCSPs’ fucose content, while samples from northern waters (Awur Bay/AB and Pari Island/PI) had a high amount of total sugar and crude fiber. Principle component analysis (PCA) result on lipid-soluble components showed different tendency that SHB and AB samples were located at close proximity and characterized by larger blade size, higher content of chlorophyll, fucoxanthin, carotenoid, PUFA, total n-3 fatty acids, total n-6 fatty acids, and also lower ratio of n-6 to n-3. Results of second research stage showed that treatment of RAW 264.7 macrophage cells with UGB and AB crude lipid extracts (12.5-50 μg/ml) significantly suppressed the nitric oxide production after lipopolysaccharide stimulation, both in pre-incubated and co-incubated cell culture model. Both crude lipid extracts showed strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and high ferric reducing antioxidant power (FRAP), which were amounted to 36.93-37.87 μmol trolox equivalent/g lipid extract and 681.58-969.81 μmol FeSO4/g lipid extract, respectively. AB samples was chosen for further research stage because this sample yielded the highest total lipid content with a selective anti-inflammatory effect and strong antioxidant activity. In the third stage, selected crude lipid extract was fractionated by normal-phase column chromatography, yielding three different lipid-soluble fractions, namely DCM (dichloromethane), ACT (acetone), and MeOH (methanol). The ACT fraction exerted strongest anti-inflammatory activity in both pre-incubated and co-incubated cell culture models. This was in accordance with its DPPH radical scavenging activity and its FRAP value. The anti-inflammatory effect of brown seaweed lipid-soluble fractions was most marked in the pre-incubated cell culture model. Metabolite profiling of lipid fractions was run by UHPLC-ESI-ORBITRAPMS/ MS. Orthogonal Projection to Latent Structures (OPLS) analysis was conducted to determine some features which were significantly correlated with the bioactivity of active fraction. There were 14 feature
Keyword: anti-inflammatory, antioxidant, coastal area, lipid-soluble component, Sargassum cristaefolium, UHPLC-ESI-ORBITRAP-MS/MS, water soluble component