In Bali there are several types of seaweed that has long been used as a

source of food by people. These seaweed local name are Bulung Boni (Caulerpa

spp.) and Bulung Sangu (Gracilaria spp.). However the study of total carotenoids

content and types of carotenoids of these seaweed are very limited, therefore need

to be further sudy. The types of carotenoids in this study identified based on the

retention of value (Rf) on thin layer chromatography. This research concluded that

total carotenoids of Bulung Boni (Caulerpa spp.) higher than Bulung Sangu

(Gracilaria spp.). Total carotenoids were 57.734 mg / 100 g in Bulung Boni, and

1.776 mg /100 g in Bulung Sangu. The separation of carotenoids Bulung Boni and

Bulung Sangu on thin layer chromatography obtained several types of carotenoids.

Based on the calculation Rf values on Bulung Boni found as many as nine types of

carotenoids such as neoxanthin, astaxanthin free, antheraxanthin ,canthaxanthin

,astaxanthin monoester, fucoxanthin, chlorophyll b, astaxanthin diester, and beta

carotene In Bulung Sangu found eight types of carotenoids, such as neoxanthin,

violaxanthin, astaxanthin free, antheraxanthin, lutein, chlorophyll b, chlorophyll a,

and beta caroten

Ketut Sri Marhaeni , Julyasih

English

UPN Jatim Repository

                                                                                                                                                145                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  CAROTENOIDS  CONTENT OF COMMERCIAL SEAWEED IN BALI  K. Sri Marhaeni Julyasih  Agriculture Faculty, University of Pembangunan Nasional  “Veteran” East Java Email : smjulyasih@gmail.com   ABSTRACT In Bali there are several types of seaweed that has  long  been used as a source of food by people.  These seaweed local name are Bulung Boni (Caulerpa spp.) and Bulung Sangu (Gracilaria spp.). However the study of total carotenoids content and types of carotenoids of these seaweed are very limited, therefore  need to be further sudy. The types of carotenoids in this study identified based on the retention of value (Rf) on thin layer chromatography. This research concluded that total carotenoids of Bulung Boni (Caulerpa spp.) higher than Bulung Sangu (Gracilaria spp.). Total carotenoids  were 57.734 mg / 100 g  in Bulung Boni, and 1.776 mg /100 g in Bulung Sangu. The separation of carotenoids Bulung Boni  and Bulung Sangu  on thin layer chromatography  obtained several types of carotenoids. Based on the calculation Rf values on Bulung Boni found as many as nine types of carotenoids such as neoxanthin, astaxanthin free, antheraxanthin ,canthaxanthin ,astaxanthin monoester, fucoxanthin, chlorophyll b, astaxanthin diester, and  beta carotene In Bulung Sangu found eight types of carotenoids, such as  neoxanthin, violaxanthin,  astaxanthin free, antheraxanthin, lutein, chlorophyll b, chlorophyll a, and beta carotene Keywords : Caulerpa spp., Gracilaria spp., and  carotenoids  INTRODUCTION    Seaweeds are nutritious because of their high protein content and high concentration of minerals, trace elements, vitamin, and carotenoids. As precursors of vitamin A, carotenoids are fundamental components in our diet, and play additional important roles in human health. Industrial uses of carotenoids include pharmaceuticals, food supplements, animal feed additives, and colorants in cosmetics (Mann et al., 2000; Thomas, 2005). Carotenoids are synthesized in all photosynthetic organisms, some bacteria, and fungi (Stahl et al., 2002).  Animals are unable to synthesize them de novo, they must obtain them by dietary means (Deming et al., 2003; Liu, 2004). Carotenoids content and composition in plants can improve their nutritional value, and provide a new source for valuable materials for industry. Some carotenoids  have also been shown as potent antioxidant, inhibitor of lipid peroxidation, anti stress, anti inflammatory, and anti aging (Siems et al.,  2002; Ronen et al., 2005). Antioxidants are molecules which can safely interact with free                                                                                                                                                 146                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  radicals and terminate the chain reaction before vital molecules are damaged. Although there are several enzyme systems within the body that scavenge free radicals, the principle micronutrient (vitamin) antioxidants are vitamin E, beta-carotene, and vitamin C. The body cannot manufacture these micronutrients so they must be supplied in the diet (Salganik, 2001).  In Bali, there are two types of seaweeds that have been consumed as vegetables. These seaweeds are Bulung Boni (Caulerpa spp.), and Bulung Sangu (Gracilaria spp.). People in Bali has  long  been used these seaweed as a source of food , but untill this time the study or publication about the total carotenoids content and types of carotenoids Bulung Boni and Bulung Sangu are very limited, therefore needs to be further study.   MATERIALS AND METHOD 1. Collection of seaweeds  Seaweeds were collected from Serangan Beach in Bali, and  dried without sunlight. 2. Analysis  of  total carotenoid Dried seaweed grinded become powder. Aproximately 0,5 g samples in powder put into centrifuge tube, added 5 ml aceton and petroleum ether, centrifuge in 3000 rpm for five minute. Take supernatant, and put into new reaction tube. The lisat added 5 ml aceton and 5 ml petroleum ether, do it for twice. Supernatant put into separate tube, and wash with destilled water. Take supernatant, and put into microtube, added 1 g Na2SO4, and then vortex. After this step take supernatant, added petroleum ether until volume 10 ml. Read absorbance in λ 450 nm with PE  as blanco.  Total carotenoid  =    3. Types of carotenoids Bulung Boni (Caulerpa spp.) and Bulung Sangu         (Gracilaria spp.)  Dried seaweed  grinded  until smooth and become powder then soaked  in a solution of dietilether for 24 hours. Furthermore carotene solution was evaporated  by poured into Eppendorf tubes. Carotene solution is the result of carotenoids extracted from seaweed that will be placed on a thin layer chromatography.  The analysis was performed with silica plates using  solution. The movement of total volume x absorbance x 100 (µg/100 g) 0,2 x sample weight                                                                                                                                                  147                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  carotenoids in the thin layer of silica was observed and calculated by the formula Rf by Crowe (2005) as follows:      Description: Rf = Relative to the front value           D1 = distance components          D2 = distance solvent   RESULTS AND DISCUSSION 1. Analysis of total carotenoids Analysis of total carotenoids in Bulung Boni (Caulerpa spp.) higher than Bulung Sangu (Gracilaria spp.). Total carotenoids contents in Bulung Boni is 57.734 mg / 100 g  and in Bulung Sangu 1.776 mg /100 g.   2. Types of Carotenoids Bulung Boni (Caulerpa spp.) and Bulung  Sangu        (Gracilaria spp.)  The result of the separation of carotenoids Bulung Boni (Caulerpa spp.) and Bulung Sangu (Gracilaria spp.) on thin layer chromatography  obtained several types of carotenoids. Based on the calculation Rf values on Bulung Boni found as many as nine types of carotenoids such as neoxanthin, astaxanthin free, antheraxanthin ,canthaxanthin ,astaxanthin monoester, fucoxanthin, chlorophyll b, astaxanthin diester, and  beta carotene. In Bulung Sangu found eight types of carotenoids, such as neoxanthin, violaxanthin,  astaxanthin free, antheraxanthin, lutein, chlorophyll b, chlorophyll a, and beta carotene (Table 1). Bulung Boni (Caulerpa spp.) is a group of divisio Chlorophyta; Class Chlorophyceae; Ordo Caulerpales; Family Caulerpaceae; Genus Caulerpa; Species Caulerpa spp. Bulung Sangu is a group of Divisio Rhodophyta; Class Rhodophyceae; Ordo Gigartinales; Family Gracillariaceae; Genus Gracillaria; Species Gracillaria spp. Burtin (2002), reported that the main carotenoids in the red algae are β-carotene and α-carotene, and their dihydroxylated derivatives zeaxanthin and lutein. The carotenoid composition of the green algae is similar to that of higher plants. The main carotenoids present are β-carotene, lutein, violaxanthin, antheraxanthin, zeaxanthin, and neoxanthin.  Rf  = D2 D1                                                                                                                                                 148                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  Table 1.  Rf value of carotenoids from Bulung Boni  (Caulerpa  spp.)  and Bulung                 Sangu (Gracilaria spp.)  Types of seaweed Rf value Types of carotenoids Caulerpa spp. 0,09 Neoxanthin 0,30 Astaxanthin free 0,36 Antheraxanthin 0,41 Canthaxanthin 0,48 Astaxanthin monoester 0,50 Fucoxanthin 0,59 Chlorophyll b 0,70 Astaxanthin diester 0,98 Beta carotene Gracilaria spp. 0,09 Neoxanthin 0,21 Violaxanthin 0,30 Astaxanthin free 0,36 Antheraxanthin 0,42 Lutein 0,55 Chlorophyll b 0,69 Chlorophyll a 0,98 Beta carotene  There are two main clasess of naturally occurring carotenoids: carotenes, which are hydrocarbons that are either linear or cyclized at one or both ends of the molecule (such as β-carotene, α-carotene), and xanthophylls, which are oxygenated derivatives of carotenes. All xanthophylls produced by higher plants, for examples violaxanthin, antheraxanthin, zeaxanthin, neoxanthin, and lutein, are also synthesized by green algae. However specific green algae possess additional xanthophylls such as astaxanthin and canthaxanthin. In addition, diatoxanthin, diadinoxanthin, and fucoxanthin, are produced in brown algae or diatoms (Eonseon et al., 2003). Biosynthetic pathway for carotenoids are Farnesyl pyrophosphate (FPP) combining with C5- isoprenoid units is extended to C20 molecules, geranylgeranyl pyrophosphate (GGPP) by geranylgeranyl pyrophosphate synthase (crtE). The common C40 carbon results from the condensation of  two C20 molecules by phytoene synthase (crtB). The sequential desaturation steps and cyclization of the ends of the molecule to generate carotenes are catalyzed by phytoene desaturase (crtP/crtI), ζ- carotene desaturase (crtQ) and lycopene cyclase (Figure 1) (Liang, 2006). Figure 1. Show The names of enzymes are according to the crtE, geranylgeranyl pyrophosphate synthase; crtB, phytoene synthase; crtP, phytoene desaturase; crtQ, zeta-carotene desaturase; crtL, lycopene beta cyclase, ; lycopene                                                                                                                                                 149                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  epsilon cyclase; cruA, the most like candidate for lycopene cyclase by comparison to CT0456 in the species lacking crtL. crtO/crtW, beta-carotene ketolase; crtR, betacarotene hydroxylase.  Figure. 1. Biosynthetic pathway of carotenoids.  CONCLUSION Total carotenoids of Bulung Boni (Caulerpa spp.) higher than Bulung Sangu (Gracilaria spp.). Total carotenoid  are 37,249.00 µg/ 100 g  in Bulung Boni, and 1,777.63 µg /100 g in Bulung Sangu. In Bulung Boni found as many as nine types of carotenoids such as neoxanthin, astaxanthin free, antheraxanthin, canthaxanthin ,astaxanthin monoester, fucoxanthin, chlorophyll b, astaxanthin diester, and  beta carotene In Bulung Sangu found eight types of carotenoids, such as  neoxanthin, violaxanthin,  astaxanthin free, antheraxanthin, lutein, chlorophyll b, chlorophyll a, and beta carotene ACKNOWLEDGMENT  Deepest thanks are due to Prof. dr. I Ketut Suata, Sp.MK, Ph.D as the Promotor, Prof. Ir. I Gede Putu Wirawan, M.Sc, Ph.D and Prof. drh. I Nyoman Mantik Astawa, Ph.D as the Co-Promotors,  for their moral and substantial supports in finalizing this research. We would like thanks also to Prof Kazuhito Kawakita,                                                                                                                                                 150                                  Sub Theme      :  Biodiversity and Conservation                                            Oral Presentation  ISBN                :  978-602-8915-93-9   ISNAR C2FS PROCEEDING Natural Resources Climate Change and Food Security in Developing Countries Surabaya, Indonesia, June 27-28, 2011  Ph.D which has provided the opportunity to use the equipment in the  Nagoya University laboratory.  REFFERENCES Burtin, P. 2002. Nutritional Value of Seaweeds. Electronic Journal of Enviromental,   Agricultural and Food Chemistry. ISSN: 1579-4377.  Crowe, K. 2005. Plant Pigment Chemistry: Pigment Extraction and Analysis using   Thin Layer Chromatography. University of Marine.  Deming, M. D., Boileau, T.W.M., Heintz, K.H.,  Atkinson, C.A.,  and Erdman, J.W.  2003. Carotenoid: Linking Chemistry, Absorption, and Metabolism to Potential Roles in Human Health and Disease. Hand book of Antioxidants. Second Edition. Revised and Expandeed Edited by Erique Cadences Lester Packer. University of Southern California School of Pharmacy. Los Angeles California. p 189-233.  El-Baky,H.H., El-Baz, F.K., dan  El-Baroty,G.S. 2007. Production of carotenoids           from Marine Microalgae and its Evaluation as safe Food Colorant and Lowering Cholesterol Agents. American Eurasian J.Agric.&Sci.2(6): 792-800.  Eonseon, J., Polle, J.E.W., Lee, H.K.,  Hyun, S.M.,  and Chang,M. 2003.          Xanthophylls in Microalgae: From Biosynthesis to Biotechnological Mass Production and Application. J Microbiol. Biotechnol. 13(2): 165-174.  Liang,C.,  Fangqing, Z .,  Wei, W., Zhangxiao, W., Song, Q. 2006. Carotenoid            Biosynthesis in Cyanobacteria: Structural and Evolutionary Scenarios Based on Comparative Genomics. International Journal of Biological Sciences.  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CAROTENOIDS CONTENT OF COMMERCIAL SEAWEED IN BALI

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CAROTENOIDS CONTENT OF COMMERCIAL SEAWEED IN BALI

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