KR20060006416A - Environmental-friendly pollution-proof material made from dioscorea batatas - Google Patents

Abstract

The present invention relates to an environmentally friendly antifouling agent derived from wild horses, containing one or more substances selected from acetophenone, 1-octadecanol or arachadidic acid, and the present invention. It is harmless to the environment and has antifouling ability against a wide range of target organisms, and since it is extracted from natural products, it is possible to obtain cheap antifouling agents, thus effectively preventing the pollution of the marine environment caused by the use of toxic antifouling agents such as TBT. have.

Antifouling agent, engraftment, bronco, acetophenone, 1-octadecanol, arachadic acid

Description

Eco-friendly antifouling agent derived from wild horses {Environmental-friendly pollution-proof material made from dioscorea batatas}             

1 is a graph showing the effect of spore adhesion of the fractions F1-F6,

2 is an HPLC profile of fraction F2,

3 is an HPLC profile of fraction F5,

4 is a profile of the GC-Mass spectrum of acetophenone,

5 is a profile of the GC-Mass spectrum of 1-octadecanol and arachatidic acid,

6 is a profile of ¹³ C NMR spectrum of acetophenone,

7 is a profile of ¹ H NMR spectrum of acetophenone,

8 is a two-dimensional NMR spectrum of acetophenone,

9 is a profile of ¹³ C NMR spectra of 1-octadecanol and arachatidic acid,

10 is a profile of ¹ H NMR spectra of 1-octadecanol and arachadinic acid,

Figure 11 is a two-dimensional NMR spectrum of 1-octadecanol and arachatidic acid.

The present invention relates to an environmentally friendly antifouling agent, and more particularly, to an environmentally friendly antifouling agent derived from wild horse (Dioscorea batatas).

Adherent organisms at sea are attached to marine facilities, ship surface, submarine surface nuclear power plant cooling system, pier, cage network of farms, as well as farming facilities, causing problems that degrade their function. . Among the fouling organisms attached to ships or offshore structures, soft-tolerant organisms are typically Enteromorpha. Harder-tolerant organisms are mainly barnacles and mussels. These attached organisms are particularly attached to large ships such as cargo ships and oil tankers, causing increased friction and problems such as speed reduction and fuel consumption.

When the adherents adhere to the bottom of the ship, the hull surface is rough and needs to be repaired frequently, and there is a problem such as a decrease in speed. When the thickness is increased by 10 μm by the adherent organism, fuel consumption increases by 6%, and when the fouling rate is 5%, fuel consumption increases by about 10%, 12-15% by 30%, The fuel consumption is said to reach 40%. In the case of large ships, fuel costs account for 50% of the ship's operating costs, so the problem of adherent organisms on the bottom of the ship is deepening socially, culturally and economically.

 Although physical cleaning methods are mainly used for artificial cleaning, antifouling paints with antifouling agents are used due to troublesome problems. As antifouling agents, copper and tin materials are known. In the past, cuprous oxide was mainly used. Since 1970, TBT (tributyltin) having excellent antifouling effect has been mainly used. Organotin compounds, including TBT, are widely used as PVC stabilizers, various plastic additives, industrial catalysts, pesticides, fungicides, wood preservatives, and the like. In particular, TBT, which is added to marine paints, has been widely used since the 1970s due to its high adhesion barrier effect. Conventionally, ship paints with copper were used, but they were replaced by TBT, which has a superior adhesion barrier effect, and TBT began to be widely used as an anti-adhesive agent that prevents adherents from sticking to marine structures and fishing nets as well as ships. did.

Although TBT compounds are added to antifouling paints due to economic problems, these materials affect non-target organisms, causing fatal problems such as reduction of aquatic organisms and accumulation in organisms. In other countries, reduced yields of oyster farms can be found in poor harvesting due to TBT contamination in coastal waters. TBT is a very toxic compound and concentrates on fish and shellfish. It causes problems such as fertility inhibition, development inhibition, and motility deterioration depending on the concentration, and when the pollution is deepened, the shell of the oyster is malformed and slowed down. In particular, gastropods such as goose, periwinkle, and conch cause imposes.

To date, research has demonstrated that TBT not only affects the killing and mortality of shellfish larvae, but also causes deformation such as oyster growth slowing and thickening of shells. In addition, the acute toxic effects of TBT on fish have been reported to vary from 1.5 to 240 μg / L, although the results vary considerably between researchers. Subsequent toxicity studies have shown that TBT causes oyster growth inhibition and shell malformation, affects the growth of plaques, reduces the growth rate of mussels, and induces high mortality of mussel larvae. In addition, it has been found that the gastrointestinal tract causes impulse sex, which is used as a useful biomarker of TBT contamination, and it is known that the impulse sex is induced even at a TBT concentration of 1 ng / L or less in seawater. The toxicity of organotin compounds is related to the number and nature of the organic groups. Triorganotin is particularly toxic and TBT is the most toxic. TBT is mainly used for antifouling paints, and TPT (triphenyltin) is used together to increase the effect.

The damage caused by TBT was first reported in France, which was hit hard by the oyster industry. The use of antifouling paints is regulated by the fact that it has a lethal effect on the growth and growth of many organisms. This trend banned the use of TBT in boats, and as a result, TBT enrichment was reduced to some extent and oyster production was restored, but recent studies have reported that TBT use has been suppressed, but TBT concentrated in sediments is still widespread. have. Since the Oyster Incident in France, extensive research has been conducted on the implementation of TBT on commercial shellfish. Because of this toxicity, France, the United Kingdom, Canada and the United States preferentially ban the use of ships less than 25m in length.

Recently, with the trend of prohibiting the use of TBT, development of TBT substitute materials is in progress. The currently developed antifouling candidates are chlorathalonil, irogarol, and dichlofuanoid. Kalihinene and ceratinamide have also been reported. It has been known by foreign researchers that some seaweeds also have antifouling substances. Among them, tannin from Sargassum natans , bromophenol from Rhodomela larix , diterpen (Schmitt et al. 1995) from Dictyota menstrulais, and halogenated furanone (de Nys et al. 1995) from Delisea pulchra . Known. Development of antifouling agents using secondary metabolites or natural materials continues, but no excellent eco-friendly antifouling agents have been found.

The present invention is to solve the problems described above, to provide an environmentally friendly antifouling agent that is harmless to the environment and excellent in antifouling ability. Another object of the present invention is to provide an antifouling agent having a low production cost.

The antifouling agent of the present invention contains at least one selected from acetophenone, 1-octadecanol, and arachadidic acid (Eicosanoic acid).

Hereinafter, the present invention will be described in detail.

The inventors have tested the antifouling ability against various algae and terrestrial plants to develop antifouling agents that are harmless to the environment. Anti-adhesion effect was tested on Ulva pertusa , one of soft-attached seaweeds. The scallions were collected from Gyeongpodae near Gangneung and transported to the laboratory to classify only the scallops, and then repeated 3 times of ultrasonic treatment for 1 minute to remove adherents, and then washed with sterilized seawater and used for the experiment.

Experimental results showed that the anti-adhesion effect of wild horses was excellent. Among the wild horse extracts, the antifouling activity components were acetophenone, 1-octadecanol, and arachadidic acid. Became me.

Dioscorea batatas DECNE, a terrestrial plant, is a perennial herbaceous herb that grows lightly and roots in the ground. Stems are purple, ridged, thin and gathered, and several branches are stretched out by winding others. The leaves are heart-shaped, close to triangles, thick and leathery, facing the stem vines, and the long petioles are often light with leaf veins, and granules are formed on the leaf axles. The leaves face each other or turn three. The shape is ovoid, 3-7㎝ long, 2-4㎝ wide, and there are many changes in the shape. In June and July, water flowers bloom straight and male flowers fall down. The stem roots are columnar thickened and go deep into the ground to form a long, large mass. The outer shell is grayish brown and easily broken into fiber-free flesh. Tuber is club-shaped, over 1m long, soft and sticky. Kinds of plants include doco roma, maple and round yam. Peeled skin and dried as it is a mother medicine (毛 山藥), the thickest of these is called water to soften and then pushed on a flat plate to process a uniform column is called mining medicine (光 山藥).

In Chinese medicine, horses and yams are called antacids and are effective in treating physical weakness, pulmonary tuberculosis, lack of energy, nocturnal enuresis, diarrhea, diabetes, hypohidrosis, and urine frequently. As the heavenly dragon is used for arthritis, back pain, bruises, relieve, and asthma. Hemp contains a lot of starch and protein and is rich in vitamin C. Medicinal ingredients include amylose, alaginine, yonogenin, saponin, dioscin and mucin. mucin, allatoin, choline, batatasin, amylase and amino acids (Seong, 1996). The steroids in wild horses are used to produce birth control pills and sex hormones in modern medicine. Cortisone, an arthritis drug, is also known as a corticosteroid derived from wild horses.

Embodiments of the present invention are as follows.

(Example 1)

Sampling and Extraction of Samples

Wild horses ( D. batatas ) were collected from Andong, Jinyang, and Yeongpung provinces to remove this material, washed, dried and crushed to make powder of wild horses 30kg. Put 30kg of sample powder in 50ℓ of methanol, store it for 24 hours, extract, collect only the supernatant, combine them three times, combine them, and evaporate methanol to 10% with a reduced pressure concentrator at 37 ℃, and filter with 0.22 ㎛ filter. It was used for the experiment while storing at -20 ℃.

(Example 2)

1st fraction

The extract obtained in Example 1 was fractionated into five fractions (A-E) by the following method using silica gel chromatography.

Silica gel (70-230 mesh) was filled under hexane, and a glass tube column (10 cm x 90 cm, PYREX) was used according to the amount of the sample, and the amount of silica gel was 50-60 times the amount of the sample. The main developing solvents were [hexane, hexane: ethyl acetate = 95: 5, hexane: ethyl acetate = 90: 10, hexane: ethyl acetate = 85: 15, hexane: ethyl acetate = 80: 20, hexane: ethyl acetate = 70 : 30, Hexane: ethyl acetate = 60: 40, Hexane: ethyl acetate = 50: 50, Ethyl acetate, Ethyl acetate: Methanol = 95: 5, Ethyl acetate: Methanol = 90: 10, Ethyl acetate: Methanol = 80: 20 ], And sequentially fractionated at a flow rate of 5 ml / min. Thin layer chromatography was carried out under the same developing solvent conditions and fractionated into six fractions.

The experiment on the inhibition of spore motility was carried out on the six fractions (I-VI) fractionated, and the secondary silica gel column was performed after combining I and II among the active fractions. Hexane: ethyl acetate = 95: 5, hexane: ethyl acetate = 90: 10, hexane: ethyl acetate = 85: 15, hexane: ethyl acetate = 80: 20, hexane: ethyl acetate = 70: 30, hexane: ethyl acetate = 60:40, hexane: ethyl acetate = 50: 50, ethyl acetate, ethyl acetate: methanol = 95: 5, ethyl acetate: methanol = 90: 10, ethyl acetate: methanol = 80: 20, sequentially 3 ml / Aliquots were made at a flow rate of min. Thin layer chromatography was carried out under the same developing solvent conditions and fractionated into five fractions.

(Example 3)

*Inhibitory effect of spore motility on punctured red seaweed

The anti-adhesion effect of terrestrial plants and algae extracts was tested on Ulva pertusa , one of the soft soft seaweeds. The scallions were collected from Gyeongpodae near Gangneung and transported to the laboratory to classify only the scallops. After sonication was repeated three times for 1 minute to remove adherents, they were washed clean with sterile seawater. After immersion in 1% betadine and 2% trition X-100 mixed solution for 1 minute, the solution was semi-dried after simple sterilization.

The semi-dried green seaweed was placed in sterile seawater and spores were released in a 80μmol / m ² s, 20 ℃ incubator. 10 μl of dimethyl sulfoxide (DMSO) was added to the prepared tube, and spore releasing seawater was added to determine the effect of spore motility according to the dilution concentrations of 500, 1000, 1500, 2000, and 4000 μl under microscope (Olympus CK-2). Assay at magnification. ++++ if no motility was observed (100%), +++ for 95% -75% spore motility, ++ for 75% -50%, 50% -20 % Was indicated as +, and when no spore motility inhibitory effect was indicated as-. Before inoculation at each dilution concentration, the movement state of the spores in seawater was checked and used in comparison with the control group.

Spore motility inhibitory activity test was carried out on the five fractions (A-E) fractionated by Example 2, the results are shown in Table 1 below.

         Conc. (Μg / ml) Activity A B C D E 4,000 ++++ ++++ ++++ ++++ ++++ 2,000 ++++ ++++ ++++ ++++ ++++ 1,000 ++++ ++++ ++++ ++++ ++++ 500 ++++ ++++ ++++ ++++ ++++ 250 ++++ +++ +++ +++ +++ 125 ++++ +++ +++ +++ +++

As confirmed in Table 1, the fraction A showed the most excellent effect of inhibiting the motility of the spores.

(Example 4)

* 2nd fraction and antifouling test

Fraction A was applied to a high-performance liquid chromatography C18 reversed phase column (Microsorb, 21.4 mm x 250 mm) for fractionation, using a 80% methanol and 20% water mixture for 5 minutes under a stable condition. The fractions were eluted at min and the fractions were separated into six F1-F6 fractions while measuring the absorbance at 254 nm. The inhibitory effect of lamellae on each fraction showed the strongest activity in F2 and F5 (Table 2), and the spore adhesion inhibitory effect (ADM) is shown in FIG. At concentrations of 10ppm and 100ppm, fractions F2 and F5 showed 75% adhesion inhibition and 1ppm concentration showed 50% adhesion inhibition. In the fractions F1, F3, F4, F6 did not show the effect of spore adhesion.

Conc. (Μg / ml) Activity F1 F2 F3 F4 F5 F6 4,000 ++++ ++++ ++++ ++++ ++++ ++++ 2,000 ++++ ++++ ++++ ++++ ++++ ++++ 1,000 +++ ++++ +++ ++++ ++++ ++++ 500 +++ ++++ +++ ++++ ++++ ++++ 250 +++ ++++ +++ +++ ++++ +++ 125 + ++++ ++ +++ ++++ ++

(Example 5)

Liquid / Gas Chromatography Mass Spectrometry (HPLC / GC Mass Spectrum)

Fractions fractionated using high performance liquid chromatography for fractionation were applied to a high performance liquid chromatography C18 reversed phase column (Microsorb, 4.6 mm × 250 mm, Cosmosil), where 80% methanol and 20% water solvents were used under stable conditions. Elution was carried out at 1.0 ml / min for 60 minutes using, and the fractions fractionated while measuring absorbance at 254 nm were shown (FIGS. 2 and 3). High performance liquid chromatography (HPLC) recovers only the active part (methanol concentration 80%), dries it, dissolves in methanol and applies 1 mg to gas chromatography mass spectrometry (Hewlett-Packard, 30 m × 0.25 mm × 0.25 μm). As a mobile phase, helium was used, and the injection ratio was 0.6 mL / min, which was analyzed using the split injection method (1:50).

Molecular weights were determined using liquid / gas chromatography mass spectrometry spectra (LC, GC / MS) for F2 and F5. The analysis results of gas chromatography mass spectrometry spectra were identified as Rt: 2.8-7.163min, 71.56min Molecular ion: M + -369, -342.

(Example 6)

* Nuclear Magnetic Resonance

As a result of analysis by hydrogen-nuclear magnetic resonance and carbon-nuclear magnetic resonance for F2 and F5 fractionated using high performance liquid chromatography (FIGS. 6-11), the ¹H NMR (500 MHz, CDCl₃ / TMS) spectrum of F2 was obtained. At δ2.05, a methyl group based on a single-signal carbonyl group is shown, possibly representing an acetoxy methyl propane group, and at δ7.53 and δ7.72 four aromatic protons in complex form. F2 was aromatic keto, ¹³C NMR spectra were found at δ171.4 at acetoxycarbons, and other aromatic carbons at δ18-131. Based on these data, F2 was identified as acetophenone (Equation 1), and the F1 ¹H NMR (500 MHz, CDCl₃ / TMS) spectrum showed six methyl protons in three pairs at δ0.95-1.03. At δ 1.25 methylene protons are shown as a single signal. Also in δ 3.45, complex signals are shown as hydroxyl protons and hydroxyl acids.

Taken together, these data turn out to be complex long chained alcohols and acids. The ¹³C NMR spectrum shows methyl, methylene and acedic carbonyl groups at δ 175.5. Analysis of all data revealed that F5 was a mixture of 1-octadecanol and arachadic acid (Eicosanoic acid) (Equations 2 and 3).

The present invention is harmless to the environment, has antifouling ability against a wide range of target organisms, and since it is extracted from natural products, it is possible to obtain an inexpensive antifouling agent, thereby effectively preventing the pollution of the marine environment caused by the use of toxic antifouling agents such as TBT. can do.

Claims (1)

Eco-friendly antifouling agent derived from wild horses containing at least one selected from acetophenone, 1-octadecanol or arachadidic acid.

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