KR20020017653A - Antifouling agent extracted from marine algae - Google Patents

Abstract

PURPOSE: Provided is an antifouling agent using the extract of a seaweed, which is friendly to environment and shows a strong antifouling activity against Enteromorpha prolifera, Mytilus edulis, etc. and replaces tributyl tin(TBT). CONSTITUTION: The antifouling agent contains the fraction III and the fraction IV produced by a process comprising the steps of: adding about 10ml of a methanol-water(4:1) solvent to 1g of dried Ishige sinicola powder and mixing and filtering; concentrating the extract at 40deg.C or lower; adjusting pH to be 1-2 by adding an acid; extracting the fraction III by using chloroform; adding ammonia to the remaining water solution layer to adjust pH to be 10; extracting the fraction IV by using a chloroform-methanol(3:1) solvent.

Description

Environmentally Friendly Antifouling Agent Using Extract of Seaweeds {Antifouling agent extracted from marine algae}

The present invention relates to an environmentally friendly antifouling agent using a water-soluble metabolite produced by algae, and more particularly, the present invention relates to Phaeopyta, Phaeophyceae, Chondariales, Peas ( The present invention relates to an environmentally friendly antifouling agent using a water-soluble metabolite with excellent antifouling function produced from Ishige sinicola, an algae belonging to Ishigeaceae.

Antifouling agents are components mixed with paints to prevent the attachment and growth of marine adherents (microorganisms and plants and animals) to the surface of ships. The nonuniformity of the hull surface is directly related to the frictional resistance of the ship, and when marine life adheres to the hull surface, it not only increases the corrosion of the hull, but also increases the frictional resistance of the ship, increasing fuel consumption and increasing economic losses. . Antifouling paints that have been used to prevent the attachment of marine adherents to the hull mainly contain tributyltin of organic tin type. Tributyltin has been used since the 1960s, but it is very antifouling, but it has been found to be a source of marine environmental pollution and environmental hormones such as serious damage to coastal fish and shellfish growth and sexual transformation. This has risen. Most developed countries have been banning the use of tributyltin since 1982, and ships of 25 m or less in length are banned in most countries. Antifouling agents containing copper or other metals are commercially available to replace tributyltin. However, in the long term use, marine environmental pollution and damage to non-target organisms are aggravated by the toxicity of metal ions. The use of higher concentrations of heavy metals compared to tin increases the environmental damage and economic burden. U.S. Patent No. 5,607,741 (1997.3.4) has an example of detecting Zosteric acid, an antifouling activity in a marine pizza plant, Zostera marina. Although it is known that there is insufficient antifouling function for practical use.

As the world's population continues to grow, terrestrial resources are reaching their limit, and the importance of developing and utilizing marine biological resources is increasing. In particular, antifouling agents are essential materials for the protection and development of the marine environment, but existing antifouling agents, including tin or copper, cause marine environmental pollution and have a fatal effect on marine ecosystems.

Accordingly, the present invention is to solve the above problems, an object of the present invention is a new antifouling agent that can replace tributyl tin (TBT), antifouling activity from the water-soluble metabolite of marine organisms that tend to be environmentally friendly To develop excellent and environmentally friendly antifouling agent.

1 is a process chart showing the separation of the active fractions from the dry decomposition products of the broad strip.

1) Gathering of widespread

The broad leaf belongs to Phaeopyta, Phaeophyceae, Chordariales and Ishigeaceae, and its scientific name is Ishige sinicola (Setchell et Gardner) Chihara. It is heterogeneous generation alternation, and frond is composed of milk tissues formed by attaching to other cell lines. The stellate is lobes and the lobes are branched. The frond is blackish yellow in color and has a slimy discharge on the surface of the frond. Habitats inhabit tropical and temperate intertidal zones.

From October 1998 to July 2000, Ishige sinicola was collected from Korea's coastal coasts (Sokcho, Daecheon, Geoje, Wando, Busan, Seongsanpo). The collected seaweed fronds were removed by using a soft brush, washed with clean fresh water, completely dried at room temperature, and then ground.

2) Test of antifouling activity

The target organisms for confirming the antifouling activity of the broad-leafed shells were Enteromorpha prolifera, which is widely distributed and frequently attached algae, and mussels (Mytilus edulis).

Spiny seaweed was collected from the seaweed farms of Korea (Gimhae) coast and mussels were collected from Korea (Busan) coast. For thorny phalaenopsis, the growth rate and spore engraftment rate of fronds were examined, and for mussels, the degree of repulsion of foot and engraftment of mussel larvae were checked. Broad strip was used after crushing completely dried at room temperature for 3-7 days. 50 ml of methanol was added to about 1 g of the ground crushed product, extracted at room temperature for about a day, and concentrated. 40 mg of the concentrate was dissolved in 1 ml of methanol and used for testing (methanol fraction). The residue extracted with methanol was extracted with methanol and the same amount of distilled water for 1 day and then concentrated. Aqueous concentrate was also dissolved in 1 ml of distilled water and used for the test (aqueous fraction).

3) Isolation and analysis of antifouling substances

After collecting the dried honeycomb was completely dried and pulverized and the active ingredient of the honeycomb was separated through the process as shown in FIG. About 10 ml of methanol-water (4: 1) mixed solvent was added to 1 g of the dried powder of the broad leaf, triturated for about 5 minutes, and filtered. The remaining solid material was extracted using 5 ml of ethyl acetate, and the ethyl acetate extract was used as fraction II. The remaining residue was defined as Fraction I. The extract extracted with a mixed solvent of methanol-water (4: 1) was concentrated to about 1/10 (40 ° C. or less), adjusted to pH 1-2 by adding 2M sulfuric acid, and extracted three times with chloroform (5 mL of CHCl ₃ ). It was set to III. The remaining aqueous layer was brought to pH 10 with ammonia water and extracted twice with a mixed solvent (5 ml) of chloroform-methanol (3: 1) to obtain fraction IV. The remaining aqueous solution was fraction V. For each fraction

Fractions with antifouling activity were selected through the growth rate of spp. Spores and the rebound test of mussels.

Experimental Example 1. Experiment of growth rate of spiny blue

The fronds of prickly lanceolate are cut into 5 mm in size and incubated in PES (Provasoli's enrichment, Jap. Soc. Plant Physiol, 1968, 63-75) for 15 days to determine the optimal growth time. The size of prickly pear was measured at 2 days intervals, and the maximum daily growth rate was 12.1 ± 2.7% at day 7, and the length was 9.2 ± 0.7㎜.

To the PES culture, a methanol fraction or an aqueous fraction of a broad leaf was added to 200 µg / ml, and the growth rate of spiny blue was tested as described above. After culturing for 7 days each, the length was measured, and the broad strips showed strong antifouling activity in the methanol fraction, and the results are shown in Table 1 below.

Experimental Example 2. Spore growth rate experiment of spiny blue

Spore growth rate of spiny blue was tested by the method of Fletcher (Int. Biodeterior, 25, 407-422). The methanol fraction of the broad leaf showed a very strong engraftment effect and the results are shown in Table 1 below.

Antifouling Effect of Spinach Extracts on Spiny Blue division Growth Rate Test (%) Spore growth rate test (%) Control 100 100 Methanol extract 25 ± 11 23 ± 2 Water soluble extract 111 ± 56 69 ± 4

Experimental Example 3. Repulsion experiment of mussel foot

Repulsion experiment of mussel foot was performed according to the method of Hayashi & Miki (J. mar. Biotechnol. 4, 127-130). Mussels were cultured mature ones of 4.5 ± 0.2 cm in size. The repulsion test was carried out by diluting 1 ul of broadleaf extract (40mg / ml) in 10-fold seawater, and the repulsive effect of mussel hair was significantly increased in methanol extract of broadleaf. The results are shown in Table 2 below.

Experimental Example 4. Embryonic experiment of mussel larvae

The engraftment of mussel larvae was tested according to the method of Kitamura (Noppon Suisan Gakkaishi, 58, 75-78). Whatman paper was used as the adhesion substrate and methanol fractions were adsorbed on Whatman filter paper by concentration. The filter paper was immersed in seawater containing the larvae of mussels for 4 days to check the engraftment degree. The methanol fraction of the broad leaf completely inhibited the engraftment of mussel larvae at concentrations of 0.8 mg / ml or more. The results are shown in Table-3, and the growth rate is the ratio of the number of larvae to the total number of larvae in the seawater used.

Antifouling Effect of Mussels Extracts on Mussels division Repulsion test Control 0 Methanol extract 100 ± 0 Water soluble extract 20 ± 1

Growth rate of mussel larvae Methanol fraction concentration (mg / ml) Growth rate of mussel larvae (%) 0.0 85 0.05 62 0.2 47 0.4 29 0.8 0 1.0 0

Experimental Example 5. Separation of Antifouling Materials

The dry fraction of the broad leaf was separated by the process as shown in FIG. Fraction I to Fraction V were subjected to spore engraftment test and mussel foot repulsion experiment of spiny blue. The results are shown in Table 4 below.

Antifouling Activity of Broaded Fraction Fraction Constellation Blue spore engraftment Repulsion rate of mussel foot (%) amount % Growth I Brown fibrous solid 134.33 ± 7.57 161 0 Ⅱ True Blue Dendritic Semi-Solid 24.33 ± 8.50 29 10 ± 5 Ⅲ Dark brown solid 16.67 ± 2.89 20 50 ± 5 Ⅳ Dark blue solid 37 ± 9.85 44 100 Ⅴ Light brown solid 33 ± 8.66 40 15 ± 10 Control 83.33 ± 13.37 100 0

As described above and as can be seen through the examples, the present invention shows a strong antifouling activity against marine adherent organisms such as Enteromorpha prolifera and mussels by developing an environmentally friendly antifouling agent from algae, It can replace the TBT with problem.

Claims (5)

An antifouling composition containing an extract of Ishige sinicola. The antifouling composition according to claim 1, wherein the extract of the broadleaf is a methanol extract. The antifouling composition according to claim 2, wherein the methanol extract is a methanol fraction having a concentration of 0.1 mg / ml to 4.0 mg / ml. The dried powder of broadleaf was crushed by adding a mixed solvent of methanol and water, filtered, extracted with a mixed solvent of methanol and water, concentrated at 40 ℃ or less, acid was added to adjust pH to 1 ~ 2 and fractions extracted with chloroform (fraction III). Antifouling composition containing a. The dried powder of broadleaf was crushed and filtered by adding a mixed solvent of methanol and water, extracted with a mixed solvent of methanol and water, the aqueous layer was added to a pH of 10 with ammonia water, and then extracted with a chloroform-methanol mixed solvent (fraction IV). Antifouling composition.