KR100621357B1 - Environmental friendly pollution-proof material and its use and the manufacturing method thereof - Google Patents

Abstract

The present invention relates to an environmentally friendly antifouling material, its use, and a method for manufacturing the same, wherein the antifouling material of the present invention is an antifouling material 6-formyl-3,4-dihydro- separated from Saragassum confusum. 2,8-dimethyl-2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2H-1-benzopyran, harmless to the environment, antifouling against a wide range of target organisms In addition to exhibiting effects, since it can be extracted from nature, it is cheaper to produce than conventional antifouling substances.

Almond hat, antifouling substance, eco-friendly, green spore, antifouling agent

Description

Environmental friendly pollution-proof material and its use and the manufacturing method             

Figure 1 is a TLC diagram at 365nm of the hexane extract of the almond kernel,

FIG. 2 is a TLC diagram at 2545 nm of hexane extract of Alk.

3 is an HPLC profile for the V-e-K7 fraction,

Figure 4 shows the GC-Mass spectrum of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" Profile,

5 is a ¹ H NMR spectrum of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" Profile,

Figure 6 shows a ¹³ C NMR spectrum of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" Profile,

Figure 7 is a 2D dimension NMR spectrum of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonane) -2,8-dimethyl-2H-chroman-6-al" ego,

8 is a HMBC correlation diagram of the antifouling material of the present invention,

Figure 9 is a partial pattern by the mass of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" Is an illustration of

10 is a profile of the IR spectrum of "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" .

The present invention relates to an environmentally friendly antifouling material and its use, and a method of manufacturing the same, and more particularly, to an environmentally friendly antifouling material extracted from natural materials, its use, and a method of manufacturing the same.

Antifouling material refers to a material mixed with paint to prevent marine adherents (microorganisms and plants) from growing on the surface of the ship. The engraftment refers to the growth and attachment of benthic organisms to artificial objects. When benthic organisms adhere to the surface of a ship, economic losses occur due to the increase of frictional force, slowing the ship, promoting corrosion, and increasing fuel use.

If the bottom of the ship is exposed to seawater for 6 months, the amount of attached organisms is about 150kg / m ² In large ships, it has been reported that the friction force increases by 0.3-1.0% for every 0.01mm roughness due to the attachment of the living organisms to the hull surface, and the speed decreases by 50% due to the increase of the friction force. Done.

To solve this problem, tributyl tin (hereinafter referred to as "TBT") has been used as an antifouling agent, but TBT has been found to have an adverse effect on the marine environment. The use of TBT, which has been used as an antifouling agent since January 1, 2003, has been adopted by the Marine Environment Protection Committee (MEPC), which is responsible for the safe operation of IMO) and the prevention of marine pollution, by adopting a risk resolution on organic tin under the regulation of marine antifouling systems. It has been banned entirely, and from 2008 onwards, regulations have been enacted that ships should be completely removed from TBT.

Non-tin antifouling agents currently being used are copper oxide or zinc, as disclosed in Korean Patent Laid-Open No. 2001-0099049, and technically, non-tin antifouling paints have insufficient antifouling effect against seaweed paré. It is also expected that the use of nitrous oxide antifouling agent will be banned between 2006 and 2008 because it will accumulate in the ocean and adversely affect the environment. Therefore, it is urgent to develop an antifouling agent which is excellent in antifouling effects and environmentally friendly. It is true.

The present invention has been made to solve the above problems, and an object thereof is to provide an antifouling material which is harmless to the environment. Another object of the present invention is to provide an antifouling material having an antifouling effect against a wide range of target organisms. Another object of the present invention is to provide an antifouling agent having a low production cost.

The antifouling material of the present invention is a material isolated from the algae Saragassum confusum, 6-formyl-3,4-dihydro-2,8-dimethyl-2- (3 ', 6') by IUPAC nomenclature. It is a new substance which can be named -dienyl-8'-hydroxy-4'-methylnonane) -2H-1-benzopyran, and has a structure as shown in the following formula (1).

In the method of the present invention, the algae Saragassum confusum powder is extracted with one solvent selected from hexane, ethyl acetate or methanol, and then the supernatant is concentrated under reduced pressure to give 6-formyl-3,4-dihydro-2, And 8-dimethyl-2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonane) -2H-1-benzopyran.

Hereinafter, the present invention will be described in detail.

Sample Collection and Extraction

The algae hatban used as a sample was collected in Pohang, Gyeongsangbuk-do (36 ° latitude, 129 ° 30 'longitude) to remove foreign substances and to dry. In order to increase the extraction yield, the resultant was pulverized using a pulverizer and used as a test material. 1 kg of sample powder was added to 10 l of hexane, stored for 24 hours, extracted, and only the supernatant was collected and combined three times. After evaporating and removing hexane by 1/10 using a vacuum condenser at 37 ° C, the resultant was filtered through a 0.22 µm filter. It was then used for experiments while stored at -20 ° C.

-Check the solubility in organic solvents

In order to determine the solubility of organic alginate hexane extracts in organic solvents, hexane, chloroform, methylene chloride, ethyl using thin layer chromatography (TLC) (SIL G / UV254, 0.25 mm layer with fluorescent indicator, Machereynagel Co. Germany) The solubility of acetate, ethanol and methanol was confirmed by UV lamp (UVGL-58, UV-254 / 366 nm, UVP Inc. USA) (FIG. 1,2).

-Inhibitory effect of spore motility of hole crab

Anti-adhesion effect of terrestrial plants and algae extracts was tested in Ulva pertusa, one of the typical adherent seaweeds. The forked leeks were collected from Gyeongpodae near Gangneung and transported to the laboratory to sort only the forked leeks. After sorting, sonication was repeated three times for 1 minute to remove adherents, followed by rinsing with sterile seawater. Then, it was immersed in 1% betadine and 2% Trition X-100 mixed solution for 1 minute and then sterilized and then dried.

Semi-dried green seaweed was put in sterile seawater and spores were released in 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 concentration of 500, 1000, 1500, 2000, and 4000 μl under microscope (Olympus CK2). Assay

More than 95%, that is, if motility is hardly observed, it is '++++'. If the spore's motility inhibitory effect is 95-75%, it is '++++'. In the case of 50-20%, '+', and in the case of 20% or less, '-'.

Conc. (Μg / ml)  Activity I Ⅱ Ⅲ Ⅳ Ⅴ Ⅵ Ⅶ Ⅷ 4,000 +++ ++ +++ ++++ ++++ +++ ++ +++ 2,000 ++ ++ +++ ++++ ++++ ++ ++ ++ 1,000 + ++ +++ ++++ ++++ ++ + + 500 + + ++ ++++ ++++ + + + 250 - - ++ +++ +++ - - + 125 - - ++ ++ +++ - - -

As shown in Table 1, at the concentration of 4000 μg / ml, fraction V showed the strongest inhibitory activity on the motility of the green spores.

Fraction V was developed on prep-TLC using a mixed solvent of hexane: methylene chloride: ethyl acetate = 3: 1: 1 and fractionated into five fractions, V-a, V-b, V-c, V-d and V-e, respectively. The inhibitory effect of spore motility on each fraction was shown in Table 2 below.

Conc. (Μg / ml)  Activity V-a V-b V-c V-d V-e 4,000 ++++ ++++ ++++ ++++ ++++ 2,000 ++++ ++++ ++++ ++++ ++++ 1,000 ++++ ++++ ++++ ++++ ++++ 500 +++ ++++ ++++ ++++ ++++ 250 +++ +++ +++ +++ ++++ 125 ++ +++ +++ +++ ++++

As shown in Table 2, Ve showed the strongest activity (++++) at 4000, 2000, 1000, 500, and 250 μg / ml, and showed strong activity (+++) even at the concentration of 125 μg / ml. Indicated. V-e was an organic solvent (hexane, ethyl acetate, methanol) to obtain each of the three fractions according to the solubility of each solvent. The inhibitory effect of spore motility by fractions was shown in Table 3 below.

Conc. (Μg / ml)  Activity V-e n-hx V-e EA V-e MeOH 4,000 +++ ++++ ++++ 2,000 +++ ++++ +++ 1,000 ++ ++++ ++ 500 ++ ++++ ++ 250 - ++++ ++ 125 - +++ ++

As shown in Table 3, in the fractions of V-e fractionated according to solubility for each organic solvent, the fraction eluted in the solvent of ethyl acetate shows a strong spore motility inhibitory effect.

Prep-High Performance Liquid Chromatograpy

The fraction of ethyl acetate, which was solvent-distilled Ve, was applied to a high performance liquid chromatography (prep-HPLC) C18 reversed phase column (Microsorb, 21.4mm × 250mm) for fractionation, and mixed with 80% methanol and 20% water under stable conditions. The solvent was eluted at a flow rate of 5 ml / min for 60 minutes, and an aliquot of each fraction (K1-K8) was performed while measuring absorbance at 254 nm. The results are shown in Table 4 below. The K7 fractions, which showed excellent spore motility inhibitory activity by verifying the physiological activity of each fraction fraction, were analyzed by gas chromatography / gas mass spectrometer, high performance liquid chromatography / liquid mass spectrometer Magnetic resonance spectra (GC / MS, LC / MS, NMR) were used as samples for material structure analysis.

Conc. (Μg / ml)  Activity V-e-k1 V-e-k2 V-e-k3 V-e-k4 V-e-k5 V-e-k6 V-e-k7 V-e-k8 4,000 +++ ++++ +++ ++++ +++ ++++ ++++ ++++ 2,000 +++ ++++ ++ ++++ ++++ ++++ ++++ +++ 1,000 ++ ++++ ++ ++++ ++++ ++++ ++++ +++ 500 ++ +++ ++ ++++ ++++ +++ ++++ +++ 250 ++ +++ ++ +++ ++++ +++ ++++ +++ 125 + ++ ++ +++ +++ +++ ++++ +++

High Performance Liquid Chromatograpy

High-performance liquid chromatography (HPLC) C18 reversed phase column (Microsorb, 4.6mm × 250mm, Cosmosil) The mixture was eluted at a rate of 1 ml / min for 60 minutes using a mixed solvent of 80% methanol and 20% water under stable conditions, and fractionated fractions were analyzed while measuring absorbance at 254 nm.

Gas Chromatograpy Mass Spectrum

Only the active part (methanol concentration 80%) was recovered from high performance liquid chromatography, dried, dissolved in methanol, and 1 mg was used for gas chromatography analysis. HP-5 (Hewlett-Packard, 30m × 0.25mm × 0.25㎛), a capillary column, was used for gas chromatography, helium was used for the mobile phase gas, and the injection rate was 0.6 mL / min (1:50). Used. The temperature of the inlet was 230 ° C. and the initial temperature of the column was 100 ° C., and then maintained at this temperature for 2 minutes, then the temperature was raised to 150 ° C. at a rate of 4 ° C./min and the temperature was raised to 200 ° C. at a rate of 3 ° C./min. It heated up to 250 degreeC at the speed of 7 degree-C / min, and hold | maintained for 6 minutes. Ion source 70 eV was used at 280 ° C. as the electron ionization mode (Table 5).

parameter Condition column HP-5 column (30cm × 0.25mm × 0.25㎛ Injector temperature 230 ℃ Detector temperature 230 ℃ Initial temperature 100 ℃ Programming lats 4 ℃ / min to 150 ℃ 3 ℃ / min to 200 ℃ 7 ℃ / min to 250 ℃ Final temperature 250 ℃ for 6 min Flow velocity of mobile phase gas 0.6 ml / min Split ratio 1:50

Molecular weights were determined using liquid / gas chromatography mass spectrometry spectra (LC, GC / MS) for Ve-K7 (FIGS. 3, 4). ion: M + -342 was confirmed.

Nuclear Magnetic Resonance

Hydrogen-nuclear magnetic resonance ( ¹ H NMR) spectra for Ve-K7 were obtained from the Bruker AC-200 spectrometer (500 MHz), and carbon-nuclear magnetic resonance ( ¹³ C NMR) spectra were obtained from the Bruker AC-800 spectrometer (800). MHz) (FIGS. 5-8 and Table 6).

Position δC (CDC13) δH (CDC13) 2 3 4 4a 5 6 7 8 8a 9 10 1 '2' 3 '4' 5 '6' 7 '8' 9 '10' -OH -CHO 39.68 31.63 27.59 112.83 146.05 133.67 148.07 125.93 154.07 22.35 22.65 38.12 24.30 127.54 121.39 24.30 115.84 117.64 75.40 15.94 16.27-195.68 --- 2.64 (q,) 2.66 --- 6.52 (dd) --- 6.52 (dd) --- --- 2.06 (s) 1.67 (s) 1.25 (dr, s) 2,14 (m) 5.20 (t) --- 2.14 (m) 6.30 (d) 6.39 (d) 3.35 (m) 2.06 (s) 1.63 (s) 4.80 (br, s) 9.36 (s)

^{The 1} H NMR (500 MHz, CDCl ₃ / TMS) spectrum shows five backbones, two meta-pairs re-paired on one side paired with δ6.52 linked by aromatic protons, and two olefin protons δ6.35 There are three pairs of 5.21 on each side of the pair, and the four methyl protons form δ1.63, 1.67, 2.06 as their respective units. One of the four molecules is δ2.46, and benzopyran nucleus, which contains two hydrogen atoms, acts in three pairs of δ2.66 at the aliphatic position.

In addition, δ9.30 as a monolith represents an aldehyde proton, and δ4.80 broadly indicates the position of a hydroxyl proton and is exchanged for D ₂ O. δ 1.25 and 2.14 indicate the locations of a wide range of methylenic protons and the data indicate that 14, 15 and dihydro benzo pyrans are part of the mixture of prenyl side chain double bonds.

^{The 13} C NMR (800 MHz, CDCl ₃ / TMS) spectrum shows 22 carbon signal groups. The signal group of δ 195.68 represents an aldehyde, and the olefinic carbons are represented by δ 127.5, 121.3, 117.6 and 115.8, and the units δ 148.05 and 146.07 represent an aromatic nucleus and an aldehyde consisting of two pairs of meta. This cannot suggest that aldehyde is a side chain, but is an aromatic nucleus and also a fragment of mass m / e 121.

Analysis of the data found to be "2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonan) -2,8-dimethyl-2H-chroman-6-al" The partial pattern by mass was obtained (FIG. 9).

Infra Red Spectrum

Fraction fraction Ve-K7 of the sample was measured by the infrared absorption spectrum (Fourier Infrared Spectrophotometer, Mattson, USA) KBr pellet method (Cho, et al., 1987). Analysis of infrared spectroscopy spectrum for Ve-K7 (FIG. 10) shows v max (KBr) -3710, 3669 (-OH), 2972, 2933, 2865 (CH ₃ , St), 1680 (= C = O, St ), 1597, 1520 and 1474 cm ^-1 (aromatic function), and as a result of synthesizing and analyzing the above data, the antifouling material separated from Sargassum confusum has the same structure as that of Chemical Formula 1, according to IUPAC nomenclature. "6-formyl-3,4-dihydro-2,8-dimethyl-2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonane) -2H-1-benzopyran It is a new substance that can be named ".

The antifouling material of the present invention is harmless to the environment, and exhibits an antifouling effect on a wide range of target organisms, and can be extracted from nature, so that the production cost is lower than that of the conventional antifouling material.

Claims (3)

6-formyl-3,4-dihydro-2,8-dimethyl-2- (3 ', 6'-dienyl-8'-hydroxy-4'-methylnonane) -2H represented by the following formula (1) -1-benzopyran. [Formula 1]

delete The algae Saragassum confusum powder was extracted with one solvent selected from hexane, ethyl acetate or methanol, and then the supernatant was concentrated under reduced pressure to give 6-formyl-3,4-dihydro-2,8-dimethyl-2. -(3 ', 6'-dienyl-8'-hydroxy-4'-methylnonane) -2H-1-benzopyran.

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