Livestock Research for Rural Development 23 (12) 2011 Guide for preparation of papers LRRD Newsletter

Citation of this paper

Red seaweed (Kappaphycus alvarezii DOTY) from Mollucas island water as potential flavonoid resource of natural antioxidant

V Lalopua, H Purnomo*, Sukoso* and Aulani’am*

Pattimura University, Ambon Indonesia,
* Brawijaya University, Malang Indonesia


The purpose of this research was to find out the best extraction solvent from the point of total flavonoid content and antioxidant activity of Kappaphycus alvarezii. The total flavonoid content of various solvent extracts were determined by   spectrophotometer, and  calculated as  mg QE/g while its antioxidant activity was analysed using  DPPH method and calculated as %  radical scavenging activity. Total flavonoid content and antioxidant activity data were presented as means standard deviation from three replicated experiment.


The hexane extract of  Kappahycus alvarezii showed higher total flavonoid content (15.93 %) and water  extract showed lowest  content (0.187 %). The Radical scavenging activity  of  various solvent extract was  increasing  with increased solvent extract concentrations and  the  highest  radical scavenging activity  could be obtained in  5 ug/ml hexane extract.  It can be concluded that hexane extract of Kappaphycus alvarezii could be potential as a rich resource of natural antioxidant.

Keywords: Scavenging DPPH radicals, solvent extraction


Kappaphycus alvarezii as an economically important red tropical seaweeds, which is highly demanded for its cell  wall polysaccharides, is the most important source of kappa carrageenan.  This seaweed accounts for the largest consumption worldwide  (Kumar et al 2008) has  also been widely cultivated in Indonesia and becoming production of seaweed priority target within the domestication fisheries revitalitation program of  2009. Some location in Indonesia where  this program was carried out are in Lombok, Sumba, South-East Sulawesi and Mollucas islands. However the species originated from Eastern Indonesia has not been investigated scientifically


Flavonoids as  the largest group of phenolic compound are known to contain a broad spectrum of chemical and biological activities including antioxidant and free radical scavenging properties (Kahkonen et al 1999). Santoso et al (2002) and Yoshie et al (2000) reported that series of flavonoid compounds such as  catechin (e.g gallocathecin, epicathecin, catechin  gallate), flavonols and flavonol glycosides have been identified from methanol extracts of red and brown algae. Many researches also reported  that a positif correlation has been documented between total flavonoid content and antioxidant activity of different seaweeds extracts. (Athukorala et al 2006, Jimenez et al 2001).


Antioxidant are classified by the products they formed on oxidation (these can be antioxidants themselves, inert, or prooxidants), or by what happens to the oxidation products and its oxidized form may be broken down by the organism as well as how effective the antioxidants is against specific free radicals. The most common procedure for antioxidant activity measurement was 2,2 diphenyl-1-picrylhydrazyl (DPPH) assay (Kumar et al 2008). 


The purpose of this research was to find out the best extraction solvent from the point of total flavonoid content and antioxidant activity of Kappaphycus alvarezii from Mollucas island.

Materials and Methods

Collection of Samples


Kappaphycus  alvarezii were taken from the cultivation area of seaweed in Wael Village, western Seram regency, Mollucas province. Samples  at the age of 30-35 days were harvested from the area  at 33 0 C, salinity 33 0/00 and  pH 7.


Preparation Extract


Sample were washed using tap water to clean up from sand, salt and other dirt, followed by sun drying for three days until its moisture content was  27 %. The dried sample  was ground to powder using grinder and strained with diameter of 100 mm. The powder samples  (400 g) was then  put into percolator (ratio  samples and solvent used 1 : 2) and extracted  using various solvents namely hexane, chloroform, ethyl acetate, methanol and water.  


Laboratory Analysis


Total flavonoids  were determined  using colorimetric method   as described by Quettier (2000). The ability of the extracts to scavenge DPPH radicals was determined by the method of Duan et al (2006).


Statistycal Analysis


Data were expressed as means of three determination standard deviation. Data were analyzed using one-way analysis of variance (ANOVA) followed by   student t test  using SPSS system version 16.  A significant difference was considered at the level of p< 0.05.

Results and Discussion

Total Flavonoid Content


The highest  total flavonoid of K. alvarezii  could be obtained in hexane extract (15.93  %),  followed by chloroform  extract (12.93%), ethyl  acetate extract (8.92 %), methanol extract (1.43 %) and aquades extract (0.187% ) (Table 1)

Table 1: Mean  values for total flavonoid content  using hexane solvent, chloroform , ethyl acetate , methanol and aquades  of      Kappaphycus  alvarezii  




Ethyl acetate 



Total flavonoid content #  






# mg quercetin equivalent/g of dry extract

abc  Means in the same row without common letter are different at P<0.05

Total flavonoid  was determined by colorimetric method using aluminium chloride where the principle involved is aluminium chloride forms acid stable complexes with the C-4 keto group and either the C-3 or C-5 hydroxyl group of flavones and flavonols. In addition, aluminium chloride forms acid labile complexes with the ortho-dihydroxyl group in the ring A or B ring of flavonoids. (Rajanandh  and Kavitha   2010).


Scavenging Effect on 1,1-diphenyl-2-picrylhydrazyl radical (DPPH)


DPPH is a useful reagent for investigating the free radical scavenging activities of compounds. The method is based on the reduction of alcoholic DPPH solutions in the presence of a hydrogen donating antioxidant due to the formation of the non radical form DPPH-H by the reactions (Shon et al 2003).


The results (figure 1) indicated that scavenging activity of seaweed extract were depended on its concentration. It was founded that the scavenging activity of sample increased with increasing extract concentration and solvent polarity. DPPH radical scavenging activity of various extracts were in the range of 4.54% -70.31%. The highest DPPH radical scavenging activity could be obtained in 5 mg/ml hexane extract (70.31%) and the lowest one could be obtained in 0.5 mg/ml water extract (4.54%).

Figure 1. The DPPH radical scavenging activity of concentration extract of Kappaphycus alvarezii

The research result indicated that non polar compound found in extract possesses strongest  ability to scavenge DPPH radicals. In this study, the extract concentration showed positive and significant correlation with DPPH scavenging activity (R2  0.93). Futher studies will elucidate the identity of the antioxidant molecules in the hexane extract.



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Received 29 October 2011; Accepted 24 November 2011; Published 1 December 2011

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