Abstract:
A cyclodextrin derivative is an X-hydroxynaphthalene-Y-amidated cyclodextrin formed by subjecting a starting cyclodextrin to an addition reaction with A-hydroxyl-B-carboxyl naphthalane and used for measuring the contamination degree of water.

Description:
BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     This invention relates to cyclodextrin derivatives and a process for measuring a contamination degree of water by using the same, and more particularly to a process for measuring a contamination degree of water by using the cyclodextrin derivatives according to the invention to measure a contamination degree in a purifying cartridge used in a water purifier thereby to detect an exchanging time of the purifying cartridge. 
     2. Description of Related Art 
     As a method of purifying drinking water, there have been used various water purifiers. However, it is difficult to detect an exchanging time of a purifying cartridge used in such a purifier, so that it is the present condition that the lapse of constant time is the exchanging time instead of measuring the contamination degree of the purifying cartridge. 
     The inventors have made various studies with respect to the use of the fluorescent cyclodextrin derivative as a detecting substance (sensor) for measuring a contamination degree of water existing in our immediate surroundings from the old time, but satisfactory compounds could not be obtained. 
     A certain cyclodextrin derivative is selected from the cyclodextrin derivatives as examined above and dissolved in water and added with a given organic compound, which is exposed to a ultraviolet ray to measure a fluorescent intensity developed. As a result, it has been confirmed that naphthol-modified cyclodextrin tends to indicate a remarkable change in the fluorescence intensity as shown by graph showing a relation between an amount of the organic compound added and the fluorescence intensity. Now, the inventors have synthesized various cyclodextrin derivatives by changing a position of hydroxy group on a naphthol ring and made further studies in order to satisfy the above demand. 
     SUMMARY OF THE INVENTION 
     Under the above circumstance, the inventors have examined various cyclodextrin derivatives having naphthol ring and found out that novel cyclodextrin derivatives as mentioned later are excellent as a fluorescent substance. 
     According to a first aspect of the invention, there is the provision of a cyclodextrin derivative being an X-hydroxynaphthalene-Y-amidated cyclodextrin formed by subjecting a starting cyclodextrin to an addition reaction with A-hydroxyl-B-carboxyl naphthalane. 
     According to a second aspect of the invention, there is the provision of a process for measuring a contamination degree of water which comprises adding the cyclodextrin derivative defined in the first aspect of the invention as a host compound to water dissolving a slight amount of a contaminant therein and then irradiating a ultraviolet ray thereto to develop fluorescence at a given wavelength. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     The invention will be described with reference to the accompanying drawings, wherein: 
     FIG. 1 is a graph showing a change of fluorescence intensity of a cyclodextrin derivative 1 or 2 according to the invention developed when a halide is added to an aqueous buffer solution of the derivative 1 or 2 and a ultraviolet ray is irradiated thereto according to usual manner in accordance with the increase of an amount of the halide added; 
     FIG. 2 is a graph showing a change of fluorescence intensity of a cyclodextrin derivative 1 or 2 according to the invention developed when benzene is added to an aqueous buffer solution of the derivative 1 or 2 and a ultraviolet ray is irradiated thereto according to usual manner in accordance with the increase of an amount of benzene added; 
     FIG. 3 is a graph showing a change of fluorescence intensity of a cyclodextrin derivative 1 or 2 according to the invention developed when a musty substance is added to an aqueous buffer solution of the derivative 1 or 2 and a ultraviolet ray is irradiated thereto according to usual manner in accordance with the increase of an amount of the musty substance added; 
     FIG. 4 is a graph showing a change of fluorescence spectrum of a cyclodextrin derivative 1 according to the invention in a buffer solution added with chloroform; 
     FIG. 5 is a graph showing a change of fluorescence spectrum of a cyclodextrin derivative 2 according to the invention in a buffer solution added with chloroform; and 
     FIG. 6 is a graph showing a change of fluorescence spectrum of a cyclodextrin derivative 3 according to the invention in a buffer solution added with chloroform. 
    
    
     DESCRIPTION OF PREFERRED EMBODIMENTS 
     Prior to the synthesis of the cyclodextrin derivative according to the invention, the studies leading to the success of the invention will be described below. 
     The following seven organic compounds (1)-(7) are selected as a target to be detected through fluorescence.                           
     They are guest compounds included in the cavity of cyclodextrin and form a host-guest complex when the cyclodextrin derivative is a host molecule. 
     Each of the compounds (1)-(7) is added as a guest to an aqueous buffer solution of each of the following cyclodextrin derivatives 1 and 2 as a host, to which is irradiated a ultraviolet ray according to usual manner to measure a fluorescence intensity developed. The measured results are shown in FIGS. 1-3 while increasing an amount of each guest added. 
     In these figures, ΔI is a value of I−I 0 , wherein I 0  indicates a fluorescence intensity at a constant wavelength (λem) in case of adding no guest and I indicates a fluorescence intensity at a constant wavelength (λem) in case of adding the guest. Moreover, λem is 500 nm in case of the derivative 1 and 490 nm in case of the derivative 2. 
     FIG. 1 is a ΔI curve in case of adding a halide such as dichloromethane, 1,2-dichloroethane, chloroform and carbon tetrachloride, and FIG. 2 is a ΔI curve in case of adding benzene, and FIG. 3 is a ΔI curve in case of adding 2-dimethylisobroneol and geosmin. 
     seen from the graphs of FIGS. 1-3, ΔI curve tends to increase in accordance with the increase of the addition amount of each guest in case of the derivative 1, while ΔI curve tends to decrease in accordance with the increase of the addition amount of each guest in case of the derivative 2. These results mean that the fluorescence intensity increases owing to the presence of the guest in case of the derivative 1, while the fluorescence intensity decreases due to the presence of the guest in case of the derivative 2. 
     Moreover, the change of fluorescence intensity is largest in 2-dimethylisobroneol and geosmin of FIG. 3 among these graphs, and particularly the change of fluorescence intensity in 2-dimethylisoborneol is large. 
     The details of the synthesis of the cyclodextrin derivatives according to the invention will be described below. 
     The inventors have confirmed that the fluorescence intensity of the host changes through the irradiation of the ultraviolet ray when the above organic compounds exist as a guest. Furthermore, it has been confirmed from these curves of fluorescence intensity that the change of fluorescence intensity increases according as the main chain (main nucleus) of the organic compound as the guest changes from straight to ring. 
     Incidentally, the inventors have confirmed that the fluorescence intensity increases when organic contaminants are added to an aqueous solution of a relevant naphthol derivative. 
     From the above facts, the inventors have considered that a compound formed by bonding naphthol unit to cyclodextrin increases or decreases the fluorescence intensity through the addition of the guest compound and synthesized cyclodextrin derivatives as follows. 
     The cyclodextrin derivatives according to the invention are derivatives 1, 2 and 3 as mentioned later. 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Structure of 
                 Designation of 
                   
               
               
                   
                 derivative molecule 
                 compound 
                 Abbreviation 
               
             
          
           
               
                 Derivative 1: 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 6-deoxy-6-(6-hydroxy-1-naphtho- amide)-β-cyclodextrin 
                 6,1-NAβCD 
               
               
                   
               
               
                 Derivative 2: 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 6-deoxy-6-(2-hydroxy-3-naphtho- amide)-β-cyclodextrin 
                 2,3-NAβCD 
               
               
                   
               
               
                 Derivative 3: 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 6-deoxy-6-[4-(6-hydroxy-1-naphtho- amide)butylamino]-β-cyclodextrin 
                 6,1-C 4 -NAβCD 
               
               
                   
               
             
          
         
       
     
     Moreover, a hollow truncated cone used for the structure of the above cyclodextrin derivative schematically shows a steric structure of cyclodextrin and is frequently used by those skilled in the art (see  Protein, Nucleic Acid, Enzyme,  Vol. 41, No. 9, (1996) pp1407-1414). In the hollow truncated cone, an upper opening portion is a side of primary hydroxyl group and a lower opening portion is a side of secondary hydroxyl group. 
     In the above derivatives 1, 2 and 3, the position of OH may arbitrarily be changed on a naphthol ring of the cyclodextrin derivative. In the derivative 3, a compound having a carbon number of 1-10 may be used for bonding cyclodextrin to naphthol ring. 
     All of the derivatives 1-3 (1-NAβCD, 2,3-NAβCD and 6,1-C 4 -NAβCD) are soluble in water. When a ultraviolet ray is irradiated to an aqueous solution of each of these derivatives, fluorescence is developed as previously mentioned. In this case, it has been confirmed that the fluorescence intensity curve shows a peak around 500 nm. 
     Each of these derivatives 1-3 is synthesized as follows:                           
     In the above synthesis of the derivatives 1-3, 6-O-Ts-βCD, 6-aminoβCD and 6-C4-βCD as an intermediate are synthesized as follows: 
     [Synthesis of each Intermediate] 
     (a) Synthesis of 6-O-tosyl-βCD(6-O-Ts-βCD): 44 g of β-CD is dissolved in 800 ml of an aqueous solution of 3N NaOH and 22 g of p-toluenesulfonyl chloride is added to react at 0-5° C. for 40 minutes with stirring. After the completion of the reaction, the reaction mixture is filtered through a glass filter and then the filtrate is neutralized with 3N HCl to precipitate a reaction product and thereafter the resulting precipitate is recovered and dried to obtain white powder. The white powder is dissolved in about 700 ml of water under heating and insoluble matter is removed through a glass filter. After the filtrate is left to stand over a night, it is filtered to obtain 10 g of a target white crystal. This compound is identified by TLC, mass spectrum and  1 H-NMR. 
     (b) Synthesis of 6-deoxy-6-amino-βCD(6-aminoβCD): 4.4 g of 6-O-Ts-βCD is dissolved in 50 ml of water and 2.5 g of NaN 3  is added to react at 80° C. for 4.5 hours as a suspension with stirring. When the reaction solution is transparent, the reaction is stopped once and concentrated and dried by means of a rotary evaporator to obtain a white solid. The white solid is dissolved in 50 ml of dimethyl formamide (DMF) and 2.5 g of triphenylphosphine is added to react at room temperature for 5 hours with stirring and then 8 ml of a concentrated ammonia solution is added to further react for 3 hours. After the completion of the reaction, the reaction product is reprecipitated with acetone and purified through a column chromatography using CM-Sephadex C25 as a column content to obtain 2.0 g of 6-amino-βCD. The compound is identified by TLC, mass spectrum and  1 H-NMR. 
     (c) Synthesis of 6-C 4 -βCD: 3.1 g of 6-O-Ts-βCD is added with 20.0 g of 1,4-diaminobutane and reacted at 70° C. for 18 hours with stirring. After the completion of the reaction, the reaction product is reprecipitated with acetone and purified through a column chromatography using CM-Sephadex C25 as a column content to obtain 2.0 g of 6-diaminobutane βCD (6-C 4 -βCD). The compound is identified by TLC, mass spectrum and  1 H-NMR. 
     The synthesis process of each of the derivatives 1-3 using the above intermediates is as follows: 
     [Synthesis of Cyclodextrin Derivatives] 
     (a) Synthesis of derivative 1: 0.5 g of 6-amino-βCD is dissolved in 30 ml of DMF and 0.082 g of 6-hydroxy-1-naphthoic acid, 0.089 g of dicyclohexylcarbodiimide (DDC) and 0.058 g of hydroxybenzotriazole (HOBt) are added, which are stirred at 0° C. for 2 hours and reacted at room temperature for 70 hours. The reaction product is filtered and reprecipitated with acetone and further with chloroform to obtain 0.43 g of a crude product. Then, the crude product is purified through a column chromatography using Sephadex LH20 as a column content to obtain 0.30 g of light brown powder. 
     (b) Synthesis of derivative 2: The same procedure as in the derivative 1 is repeated except that 2-hydroxy-3-naphthoic acid is used instead of 6-hydroxy-1-naphthoic acid to obtain 0.14 g of a crude product. The crude product is purified to obtain 0.07 g of light beige powder. 
     (c) Synthesis of derivative 3: 0.5 g of 6-C 4 -βCD is dissolved in 13 ml of DMF and 0.086 g of 6-hydroxy-1-naphthoic acid, 0.094 g of DDC and 0.062 g of HOBt are added, which are stirred at 0° C. for 2 hours and reacted at room temperature for 22 hours. The reaction product is filtered and reprecipitated with acetone and purified through a column chromatography using CM-Sephadex C25 as a column content to obtain 0.48 g of ivory powder. 
     Confirmatory analytical values of the derivatives 1-3 are shown in Table 1. 
     
       
         
               
               
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 De- 
                   
                   
                   
                   
               
               
                 riva- 
                   
                   
                   
                 Elementary 
               
               
                 tive 
                 R f   
                 MS (m/z) 
                 NMR (ppm) 
                 analysis 
               
               
                   
               
             
             
               
                 1 
                 0.68 
                 calculated 
                 δ: 8.01(d, 1H; H-8′); 7.73(d, 1H; H-4′); 
                 rational formula: 
               
               
                   
                   
                 value: 
                 7.38(t, 1H; H-3′); 7.31(d, 1H; H-2′); 
                 C 53 H 77 NO 36 . 
               
               
                   
                   
                 [M + H] +  = 
                 7.15(d, 1H; H-5′); 7.07(dd, 1H; H-7′); 
                 5.8H 2 O.1.02DMF 
               
               
                   
                   
                 1305.8 
                 5.95˜4.75(m, 7H; H-1); 
                 calculated value: 
               
               
                   
                   
                 found 
                 3.94˜3.20(m, 42H; 
                  C = 45.40%; 
               
               
                   
                   
                 value: 
                 H-2˜6)/([D 6 ]-DMSO). 
                  H = 6.27%; 
               
               
                   
                   
                 [M + H] +  = 
                   
                  N = 1.91%; 
               
               
                   
                   
                 1308.4 
                   
                 C/N = 23.82 
               
               
                   
                   
                   
                   
                 found value: 
               
               
                   
                   
                   
                   
                  C = 45.40%; 
               
               
                   
                   
                   
                   
                  H = 6.51%; 
               
               
                   
                   
                   
                   
                  N = 1.91%; 
               
               
                   
                   
                   
                   
                 C/N = 23.80 
               
               
                 2 
                 0.68 
                 calculated 
                 δ: 8.44(s, 1H; H-4′); 7.83(d, 1H; H-5′); 
                 rational formula: 
               
               
                   
                   
                 value 
                 7.71(d, 1H; H-8′); 7.49(t, 1H; H-7′); 
                 C 53 H 77 NO 36 . 
               
               
                   
                   
                 [M + H] +  = 
                 7.33(t, 1H; H-6′); 7.24(s, 1H; H-1′); 
                 8H 2 O.0.94DMF 
               
               
                   
                   
                 1305.8 
                 4.96˜4.70(m, 7H; H-1); 
                 calculated value: 
               
               
                   
                   
                 found 
                 3.98˜3.20(m, 42H; H-2˜H-6)/ 
                  C = 44.12%; 
               
               
                   
                   
                 value: 
                 ([D 6 ]-DMSO). 
                  H = 6.26%; 
               
               
                   
                   
                 [M + H] +  = 
                   
                  N = 1.79%; 
               
               
                   
                   
                 1307.3 
                   
                 C/N = 24.66 
               
               
                   
                   
                   
                   
                 found value: 
               
               
                   
                   
                   
                   
                  C = 44.20%; 
               
               
                   
                   
                   
                   
                  H = 6.62%; 
               
               
                   
                   
                   
                   
                  N = 1.79%; 
               
               
                   
                   
                   
                   
                 C/N = 24.67 
               
               
                 3 
                 0.59 
                 calculated 
                 δ: 8.11(d, 1H; H-8′); 7.77(t, 1H; H-4′); 
                 rational formula: 
               
               
                   
                   
                 value: 
                 7.48(t, 2H; H-2, ′H-3′); 7.25(dd, 1H; H-7′); 
                 C 57 H 86 N 2 O 36 . 
               
               
                   
                   
                 [M + Na] +  = 
                 7.18(d, 1H; H-5′); 5.06˜4.95(m, 7H; H-1); 
                 4H 2 O 
               
               
                   
                   
                 1398.2 
                 3.96˜3.35(m, 44H; H-2˜5, H—CH 2 N); 
                 calculated value: 
               
               
                   
                   
                 found value: 
                 3.12(d, H-1; H-6a/b); 
                  C = 47.30%; 
               
               
                   
                   
                 [M + Na] +  = 
                 2.87(dd; 1H; H-6b/a); 
                  H = 6.55%; 
               
               
                   
                   
                 1398.5 
                 2.99(m, 2H; H—NCH 2 ); 
                  N = 1.94%; 
               
               
                   
                   
                   
                 1.8˜1.6(m, 4H; H—CH 2 )/(D 2 O). 
                 C/N = 24.42 
               
               
                   
                   
                   
                   
                 found value: 
               
               
                   
                   
                   
                   
                  C = 47.13%; 
               
               
                   
                   
                   
                   
                  H = 6.83%; 
               
               
                   
                   
                   
                   
                  N = 1.93%; 
               
               
                   
                   
                   
                   
                 C/N = 24.44 
               
               
                   
               
             
          
         
       
     
     [Properties of Derivatives] 
     The derivatives 1 and 3 are well soluble in water, while the derivative 2 is slightly difficult to be dissolved in water but is dissolved within a required concentration range in the detection. And also, these derivatives are easily dissolved by increasing pH of the aqueous solution. 
     In Table 1, TLC is carried out by using TLC plate (silica gel F254, layer thickness: 0.25 mm) made by Merck and a developing solvent comprising concentrated ammonia:ethyl acetate:2-propanol:water=1:3:5:4. 
     Naphthol unit as a light absorption unit is confirmed by irradiating a light of UV lamp to a developed plate to visually observe emission (fluorescence). Cyclodextrin portion is confirmed as a violet spot by spraying an Anis reagent (a 1:1 mixed solution of ethanol solution of 10% sulfuric acid and ethanol solution of anisaldehyde) onto the plate and heating by means of a heat gun. 
     
       
           R   f =(moving distance of derivative)/(developed distance of developing solvent) 
       
     
     The mass spectrum is measured by means of a mass spectrometer of MALDI III (TOF-MS) made by Shimadzu Seisakusho. 
     The measurement by  1 H-NMR is carried out by using Varian VXR-500S FT-NMR spectrometer. 
     [Fluorescence Intensity of Each Derivative] 
     The fluorescence spectra are recorded on a Shimadzu RF-5300PC spectrofluorophotometer. 
     Moreover, the measuring conditions are as follows. 
     The derivative 1 is dissolved in the buffer solution of 0.005M Na 2 CO 3 —NaHCO 3  with pH at 9.28 so as to have a concentration of 1.0×10 −4 M and an excitation wavelength (λex) at 25° C. is 337 nm. 
     The derivative 2 is dissolved in the buffer solution of 0.05M KH 2 PO 4 —NaOH with pH at 7.26 so as to have a concentration of 1.0×10 −4 M and λex at 25° C. is 334 nm. 
     The derivative 3 is dissolved in a buffer solution of 0.005M Na 2 CO 3 —NaHCO 3  with pH at 9.28 so as to have a concentration of 5.0×10 −4 M and λex at 25° C. is 295 nm. 
     As mentioned above, 6,1-NACD, 2,3-NAβCD and 6,1-C4-NAβCD according to the invention have hydrophilic nature and develop fluorescence when ultraviolet ray is irradiated to an aqueous solution of each of these derivatives. In this case, the fluorescence intensity curve indicates a peak around 500 nm. 
     An artificial contamination experiment is carried out by adding chloroform to the buffer solution of each of the derivatives 1-3. The experimental results are shown in FIGS. 4-6, respectively. 
     As seen from the above experiments, the contamination degree of water can easily be known by measuring the fluorescence intensity of the cyclodextrin derivative according to the invention in the contamination of water. 
     Particularly, in the purification process of city water, it is possible to optically and rapidly measure the contamination degree without taking a long time or taking various operations as in the conventional technique, so that the invention is very effective to the control of water purification process and the like.