CS274026B1 - Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation - Google Patents
Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation Download PDFInfo
- Publication number
- CS274026B1 CS274026B1 CS806788A CS806788A CS274026B1 CS 274026 B1 CS274026 B1 CS 274026B1 CS 806788 A CS806788 A CS 806788A CS 806788 A CS806788 A CS 806788A CS 274026 B1 CS274026 B1 CS 274026B1
- Authority
- CS
- Czechoslovakia
- Prior art keywords
- volume
- parts
- salt
- amino
- hydroxy
- Prior art date
Links
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 title claims abstract description 10
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 238000000034 method Methods 0.000 title claims abstract description 6
- 238000006116 polymerization reaction Methods 0.000 title claims description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 24
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000000243 solution Substances 0.000 claims abstract description 18
- 235000019441 ethanol Nutrition 0.000 claims abstract description 12
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- 239000002253 acid Substances 0.000 claims abstract description 10
- 229910052500 inorganic mineral Inorganic materials 0.000 claims abstract description 9
- 239000011707 mineral Substances 0.000 claims abstract description 9
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims abstract description 8
- 239000007864 aqueous solution Substances 0.000 claims abstract description 7
- IHDBZCJYSHDCKF-UHFFFAOYSA-N 4,6-dichlorotriazine Chemical compound ClC1=CC(Cl)=NN=N1 IHDBZCJYSHDCKF-UHFFFAOYSA-N 0.000 claims abstract description 6
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract description 5
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims abstract description 5
- 229910000027 potassium carbonate Inorganic materials 0.000 claims abstract description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims abstract description 4
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 claims abstract description 3
- -1 1-hydroxy-3,6-bissulfonaphth-8-yl Chemical group 0.000 claims description 11
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 claims description 5
- 150000001447 alkali salts Chemical class 0.000 claims description 4
- AEMOLEFTQBMNLQ-BKBMJHBISA-N alpha-D-galacturonic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-BKBMJHBISA-N 0.000 claims description 4
- 239000011734 sodium Substances 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims 1
- 239000000758 substrate Substances 0.000 abstract description 9
- 108090000790 Enzymes Proteins 0.000 abstract description 7
- 102000004190 Enzymes Human genes 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 6
- 108010059820 Polygalacturonase Proteins 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 5
- 230000002351 pectolytic effect Effects 0.000 abstract description 2
- 150000003839 salts Chemical class 0.000 abstract description 2
- 159000000011 group IA salts Chemical class 0.000 abstract 2
- 238000002679 ablation Methods 0.000 abstract 1
- 239000000975 dye Substances 0.000 description 13
- 229920001282 polysaccharide Polymers 0.000 description 6
- 239000005017 polysaccharide Substances 0.000 description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 5
- 150000004676 glycans Chemical class 0.000 description 5
- 239000001632 sodium acetate Substances 0.000 description 5
- 235000017281 sodium acetate Nutrition 0.000 description 5
- 239000012634 fragment Substances 0.000 description 4
- 229920001277 pectin Polymers 0.000 description 4
- 239000001814 pectin Substances 0.000 description 4
- 235000010987 pectin Nutrition 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 3
- 229920002230 Pectic acid Polymers 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 108010093305 exopolygalacturonase Proteins 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 2
- 239000010318 polygalacturonic acid Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical class NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 241000208125 Nicotiana Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 239000000987 azo dye Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- SCZVXVGZMZRGRU-UHFFFAOYSA-N n'-ethylethane-1,2-diamine Chemical compound CCNCCN SCZVXVGZMZRGRU-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
Vynález sa týká dvojsodnej soli 0- £4-chlór-6-£benzénazo-2-(l-hydroxy-3,6-bissulfonat-8-yl)aminoJ -1,3,5-triazin-2-ylJ D-galakturónanu o priemernom stupni polymerizácie i, a sposobu jej pripravy.The present invention relates to the disodium O-β-4-chloro-6-benzenazo-2- (1-hydroxy-3,6-bissulfonat-8-yl) amino-1,3,5-triazin-2-yl D-galacturonate. about the average degree of polymerization i, and the process for its preparation.
Rozpustné kovalentne vyfarbené polysacharidy sa v poslednej době s úspechom používajú na rýchlu a jednoduchá detekciu příslušných hydroláz polysacharidov a tiež sú vhodné na stanovenie ich aktivity / B.V. Mc Cleary: Carbohydr. Res. 86, 97 (1980); P. Biely a spol.: Anal. Biochem., 144, 147 (1985); P. Biely a spol.; 172, 176 (1988)/. Farbivo sa obvykle viaže na polysacharid priamo na hydroxylovú skupinu éterickou vazbou alebo na derivatizovaný polysacharid. Princip detekcie .a stanovenia aktivity enzýmov spočívá v zmene rozpustnosti farebného polysacharidů v dosledku hydrolýzy enzýmom a následnej tvorbě nízkomolekulárnych farebnýoh fragmentov rozpustných v přítomnosti organického rozpúšťadla. V případe pektín-degradujúcich enzýmov takýmto substrátom je kovalentne zafarbený pektín azo-farbivom N- {l-l·- [(3, 6-disulfo-l-naftyl)azoJ -naftyl-etyléndiamin, připravený D. R. Friedom a G. W. Changom / J. Agric. Food Chem., 30 (5) (1982) 982/, v ktorom je farbivo viazané na pektín cez amidickú skupinu, pri vzniku ktorej sa používá ťažko dostupný katalyzátor - vodorozpustný karbodiimid. Iný druh farebného substrátu pre pektinázy je sodná sol' 2-[ 3-(l-amino-2-sulfoantrachinón-4-ylamino)fenysulfonylJ-etyl - etyléndiamín derivátu pektínu / J. S. Huang, J. Tang: Anal. Biochem., 73, 369 (1976)/. Připravuje sa v dvoch stupňoch. Najprv sa připraví etyléndiaminoderivát pektínu a na ten sa potom viaže farbivo. Tento substrát sa však vyznačuje zlou rozpustnostíou v tlmivých roztokoch.Soluble covalently colored polysaccharides have recently been used successfully to quickly and easily detect the respective polysaccharide hydrolases and are also useful for determining their activity. Mc Cleary: Carbohydr. Res. 86, 97 (1980); P. Biely et al., Anal. Biochem. 144: 147 (1985); P. Biely et al .; 172, 176 (1988)]. The dye usually binds the polysaccharide directly to the hydroxyl group via an ether bond or to a derivatized polysaccharide. The principle of detection and determination of enzyme activity is to alter the solubility of colored polysaccharides as a result of enzyme hydrolysis and subsequent formation of low molecular weight colored fragments soluble in the presence of an organic solvent. In the case of pectin-degrading enzymes with such a substrate, the covalently stained pectin is the azo dye N- {1 - 1 - [(3,6-disulfo-1-naphthyl) azo] -naphthyl-ethylenediamine, prepared by D. R. Fried and G. W. Chang / J. Agric. Food Chem., 30 (5) (1982) 982), wherein the dye is coupled to pectin via an amide group to form a poorly available water-soluble carbodiimide catalyst. Another kind of colored substrate for pectinases is the sodium salt of 2- [3- (1-amino-2-sulfoanthraquinon-4-ylamino) phenysulfonyl] ethyl-ethylenediamine pectin derivative / J.S. Huang, J. Tang: Anal. Biochem., 73, 369 (1976)]. It is prepared in two stages. First, the pectin ethylenediamine derivative is prepared and the dye is then bound to the dye. However, this substrate is characterized by poor solubility in buffers.
Běžné techniky viazania farbíva na polysacharid, ktoré nevyžadujú jeho derivatizáciu, v případe kyseliny pektovej neposkytujú substrát s požadovanými vlastností ami. Takýmto sposobom vyfarbovania sa u kyseliny pektovej dosiahne vel'mi nízký stupeň substitúcie farbivom. Získaný substrát neposkytuje enzymatickým štiepením nízkomolekulárne farebné fragmenty, ktoré by umožňovali jeho použitie na detekciu a stanovenie enzýmovej aktivity. Enzýmom sa uvďňujú iba nízkomolekulárne nefarebné fragmenty, pričom farbivo zostáva naviazané na polymérnom zvyšku, ktorý je nerozpustný v 60 % etylalkohole.Conventional dyeing techniques dye the polysaccharide which do not require its derivatization, in the case of pectic acid, do not provide a substrate with the desired properties. In this way, a very low degree of dye substitution is achieved with pectic acid. The substrate obtained does not yield, by enzymatic cleavage, low molecular weight colored fragments that would allow its use in the detection and determination of enzyme activity. Only low molecular weight uncoloured fragments are mentioned by the enzyme, while the dye remains bound to a polymer residue which is insoluble in 60% ethyl alcohol.
Uvedené nevýhody v podstatnej miere odstraňuje vodorozpustný chromogénny substrát dvojsodná sol' O- {4-chlór-6- [benzénazo-2-(l-hydroxy-3,6-bissulfonaft-8-yl)aminoJ -1,3, 5-triazin-2-yll D-galakturónanu s priemerným stupňom polymerizácie 10 vzorca IThe above-mentioned disadvantages are substantially eliminated by the water-soluble chromogenic substrate O- {4-chloro-6- [benzenazo-2- (1-hydroxy-3,6-bissulfonaphth-8-yl) amino] -1,3,5-triazine disodium salt. 2-yl of D-galacturonan with an average degree of polymerization of 10 of formula I
Zistilo sa, že uvedenú dvojsodnú sol' O- {4-chlór-6- [benzénazo-2-(l-hydroxy-3,6-bissulfonaft-8-yl)aminoJ -1,3,5-triazín-2-yl-J D-galakturónanu s priemerným stupňom polymerizácie 10 je možné připravit' nasledujúcim sposobom podl'a vynálezu, ktorého podstata spočívá v tom, že 1 hmot. diel vodorozpustnej kyseliny oligo-D-galakturónovej o priemernom stupni polymerizácie 10 alebo jej sodnej, príp. draselnéj soli reaguje vo vodnom roztoku s obsahom 0,8 až 1,2 hmot. dielov uhličitanu sodného alebo draselného za přítomnosti alkalickéj soli kyseliny octovéj alebo minerálněj kyseliny s 0,6 až 1 hmot. dielom dvojsodnej soli 2,4-dichlór-6- [benzénazo-2-(l-hydroxy-3,6-bissulfonaft-8-yl) aminoJ -1,3,5-triazínu (dichlórtriazínové farbivo). Vzniknutý roztok zafarbeného D-gala-. kturónanu sa zneutralizuje kyselinou octovou alebo minerálnou kyselinou. 1 obj. diel zneutralizovaného roztoku sa zráža s 3,5 až 4,5 obj. dielmi roztoku obsahujúceho 2 až 3 obj.It was found that said disodium salt of O- {4-chloro-6- [benzenazo-2- (1-hydroxy-3,6-bissulfonaphth-8-yl) amino] -1,3,5-triazin-2-yl The D-galacturonan with an average degree of polymerization of 10 can be prepared by the following process according to the invention, which consists in that 1 wt. % by weight of water-soluble oligo-D-galacturonic acid having an average degree of polymerization of 10 or its sodium or sodium olefin. potassium salt is reacted in an aqueous solution containing 0.8 to 1.2 wt. parts by weight of sodium or potassium carbonate in the presence of an alkali salt of acetic acid or a mineral acid with 0.6 to 1 wt. portion of 2,4-dichloro-6- [benzenazo-2- (1-hydroxy-3,6-bissulfonaphth-8-yl) amino] -1,3,5-triazine disodium salt (dichlorotriazine dye). The resulting solution stained with D-galaxy. The urethane is neutralized with acetic acid or mineral acid. 1 vol. part of the neutralized solution precipitates with 3.5 to 4.5 vol. parts of a solution containing 2 to 3 vol.
CS 274026 Bl diely 96 %-ného etylalkoholu a 1 až 2 obj. diely acetonu. Nenaviazané dichlortriazinová farbivo sa odstráni premývaním zrazeniny roztokom obsahujúcim 2 až 3 obj. diely 96 %-ného etylalkoholu, 1 až 2 obj. diely acetonu a 1 obj. diel 0,6 až 1 %-ného vodného roztoku alkalickej soli kyseliny octovej alebo minerálnej kyseliny. Výhodné je použití na neutralizáciu takú kyselinu,ktorej sol' sa použije počas reakcie. Z hl'adiska využitia dvojsodnej soli O- £4-chlór-6- £benzénazo-2-(l-hydroxy-3,6-bissulfonaft-8-yl)aminoJ-1,3,5-triazin-2-ylJ -D-galakturónanu o priemernom stupni polymerizácie 10 ako substrátu na detekciu a stanovenie aktivity pektináz, ktoré sa uskutočňuje v octanovom tlmivom roztoku, je výhodné použití pri reakcii octan sodný a kyselinu octovú pri neutralizácii.CS 274026 B1 parts of 96% ethyl alcohol and 1 to 2 vol. parts of acetone. Unbound dichlorotriazine dye is removed by washing the precipitate with a solution containing 2-3 vol. parts of 96% ethyl alcohol, 1 to 2 vol. parts of acetone and 1 vol. % of a 0.6 to 1% aqueous solution of an alkali salt of acetic acid or a mineral acid. Preference is given to using an acid whose salt is used during the reaction. For the use of disodium O-4 4-chloro-6-benzenazo-2- (1-hydroxy-3,6-bissulfonaphth-8-yl) amino] -1,3,5-triazin-2-yl] - D-galacturonan having an average degree of polymerization of 10 as a substrate for the detection and determination of pectinase activity, which is carried out in acetate buffer, is preferably used in the reaction of sodium acetate and acetic acid in neutralization.
Výhodou dvojsodnej soli 0-£4-chlór-6-£benzénazo-2-(l-hydroxy-3,6-bissulfonaft-8-yl)amino]-1,3,5-triazin-2-ylJ -D-galakturónanu o priemernom stupni polymerizácie 10 podl*a vynálezu je, že sa ziska farebný substrát rozpustný v tlmivých roztokoch, v ktorých sa uskutočňuje enzýmová reakcia. Ďal'šia jeho výhoda je, že enzymatickým štiepením polygalakturonázou už pri prvom ataku uvol'ňuje farebné fragmenty (DP 5), ktoré zostávajú rozpuštěné v supernatante po vyzrážaní nerozštiepeného substrátu organickým rozpúšť adlom.The advantage of disodium O-β-4-chloro-6-benzenazo-2- (1-hydroxy-3,6-bissulfonaphth-8-yl) amino] -1,3,5-triazin-2-yl-β-D-galacturonate having an average degree of polymerization of 10 according to the invention is to obtain a colored substrate soluble in the buffers in which the enzyme reaction is carried out. A further advantage is that by enzymatic cleavage by polygalacturonase at the first attack, it releases colored fragments (DP 5), which remain dissolved in the supernatant after precipitation of the undigested substrate with an organic solvent.
Uvedené příklady ilustrujú, ale neobmedzujú predmet vynálezu.These examples illustrate but do not limit the scope of the invention.
Příklad 1 g kyseliny oligo-D-galakturónovej o priemernom stupni polymerizácie 10 sa rozpustí v 40 ml destilovanej vody pri teplote 30 *C a odfiltrujú sa nerozpustné podřely. Potom sa k filtrátu přidá 1 g dichlórtriazinového farbiva rozpuštěného v 6 ml vody a mieša sa 10 minút pri teplote 30 *C. Potom sa přidá 0,5 g octanu sodného a zase sa mieša 10 minút pri tej istej teplote. Nakoniec sa přidá 12 ml 6 %-ného vodného roztoku uhličitanu sodného a mieša sa 2 hodiny pri 30 *C. Reakčná zmes sa potom zneutralizuje 2 M kyselinou octovou na pH 7 a vyzráža sa roztokom obsahujúcim 120 ml 96 %-ného etylalkoholu a 120 ml acetonu. Zrazenina sa odfiltruje na nuči a premýva roztokom obsahujúcim 1 diel 0,6 %-ného vodného roztoku octanu sodného, 2 diely 96 %-ného etylalkoholu a 2 diely acetonu až do slabo sfarbeného filtrátu. Potom sa zrazenina prezráža, a to tým sposobom, že sa rozpustí v 50 ml 0,6 %-ného vodného roztoku octanu sodného a potom sa vyzráža roztokom obsahujúcim 100 ml 96 %-ného etylalkoholu a 100 ml acetonu. Premýva sa ako už bolo uvedené, až do bezfarebného filtrátu. Zrazenina sa premyje etylalkoholom, acetónom a vysuší sa.Example 1 g of oligo-D-galacturonic acid having an average degree of polymerization of 10 is dissolved in 40 ml of distilled water at 30 DEG C. and insoluble fractions are filtered off. 1 g of dichlorotriazine dye dissolved in 6 ml of water is then added to the filtrate and stirred for 10 minutes at 30 ° C. 0.5 g of sodium acetate is then added and the mixture is again stirred for 10 minutes at the same temperature. Finally, 12 mL of a 6% aqueous sodium carbonate solution was added and stirred at 30 ° C for 2 hours. The reaction mixture is then neutralized with 2M acetic acid to pH 7 and precipitated with a solution containing 120 ml of 96% ethyl alcohol and 120 ml of acetone. The precipitate is filtered off under suction and washed with a solution containing 1 part of a 0.6% aqueous solution of sodium acetate, 2 parts of 96% ethyl alcohol and 2 parts of acetone until a slightly colored filtrate. The precipitate is then precipitated by dissolving it in 50 ml of a 0.6% aqueous sodium acetate solution and then precipitating it with a solution containing 100 ml of 96% ethyl alcohol and 100 ml of acetone. Wash as above until colorless filtrate. The precipitate was washed with ethyl alcohol, acetone and dried.
Získal sa 1 g zrazeniny s obsahom farbiva 5,6 %.1 g of precipitate was obtained with a dye content of 5.6%.
Příklad 2 g Na-D-galakturónanu o priemernom stupni polymerizácie 10 sa rozpustí v 200 ml vody pri teplote 40 *C a přidá sa 5 g dichlórtriazinového farbiva rozpuštěného v 40 ml vody a mieša sa 10 minút pri tej istej teplote. Potom sa přidá postupné 2,5 g octanu sodného a 60 ml 6 %-ného uhličitanu draselného a mieša sa 2 hodiny pri 40 *C. Potom sa reakčná zmes ochladí, zneutralizuje s 2 M kyselinou octovou a zráža za stálého miešania roztokom obsahujúcim 300 ml 96 %-ného etylalkoholu a 300 ml acetonu. Zrazenina sa přefiltruje cez nuč a vymývá ako v příklade 1. Potom sa prezráža, premyje a vysuší tým istým sposobom ako v příklade 1.Example 2 g of Na-D-galacturonan having an average degree of polymerization of 10 is dissolved in 200 ml of water at 40 DEG C. and 5 g of dichlorotriazine dye dissolved in 40 ml of water are added and stirred for 10 minutes at the same temperature. Then 2.5 g of sodium acetate and 60 ml of 6% potassium carbonate are added successively and stirred for 2 hours at 40 ° C. The reaction mixture was cooled, neutralized with 2M acetic acid and precipitated with stirring with a solution containing 300 ml of 96% ethyl alcohol and 300 ml of acetone. The precipitate is filtered through suction and eluted as in Example 1. It is then precipitated, washed and dried in the same manner as in Example 1.
Získalo sa 4,8 g zrazeniny s obsahom farbiva 6 %.4.8 g of precipitate were obtained with a dye content of 6%.
Vynález može nájsť uplatnenie v potravinárskom, textilnom i tabakovom priemysle na detekciu a stanovenie aktivity polygalakturonáz v preparátoch obsahujúcich pektolytické enzýmy.The invention can be used in the food, textile and tobacco industries for the detection and determination of polygalacturonase activity in preparations containing pectolytic enzymes.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS806788A CS274026B1 (en) | 1988-12-07 | 1988-12-07 | Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS806788A CS274026B1 (en) | 1988-12-07 | 1988-12-07 | Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS806788A1 CS806788A1 (en) | 1990-08-14 |
| CS274026B1 true CS274026B1 (en) | 1991-04-11 |
Family
ID=5430785
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS806788A CS274026B1 (en) | 1988-12-07 | 1988-12-07 | Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS274026B1 (en) |
-
1988
- 1988-12-07 CS CS806788A patent/CS274026B1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| CS806788A1 (en) | 1990-08-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Signini et al. | On the preparation and characterization of chitosan hydrochloride | |
| US4814437A (en) | Method for preparation of sulfated polysaccharides by treating the polysaccharide with a reducing agent before sulfation | |
| US5442048A (en) | Process for the preparation of activated chitosans and their use in the preparation of chiotsan derivatives | |
| Hayashi et al. | Pea xyloglucan and cellulose: VI. Xyloglucan-cellulose interactions in vitro and in vivo | |
| US4970150A (en) | Process for preparing chitosan oligosaccharides | |
| Wood | Specificity in the interaction of direct dyes with polysaccharides | |
| Verraest et al. | Carboxymethylation of inulin | |
| US5733854A (en) | Cleaning compositions including derivatized guar gum composition including nonionic and cationic groups which demonstrate excellent solution clarity properties | |
| Muraki et al. | Preparation and crystallization of D-glucosamine oligosaccharides with dp 6-8 | |
| US20110218331A1 (en) | Method of Preparation of DTPA Crosslinked Hyaluronic Acid Derivatives and Modification of Said Dervivatives | |
| Somorin et al. | Studies on chitin. II. Preparation of benzyl and benzoylchitins | |
| EP1448840B1 (en) | Method for the modification of polymeric carbohydrate materials | |
| Tuzikov et al. | Conversion of complex sialooligosaccharides into polymeric conjugates and their anti-influenza virus inhibitory potency | |
| CN100560605C (en) | Nitrogen-containing polysaccharide derivatives and preparation methods thereof | |
| US4267313A (en) | Saccharide derivative having ureido group | |
| CS274026B1 (en) | Two-sodium salt of 0-(4-chloro-6-/benzenase-2-(1-hydroxy-3,6-bisulfonate-8-yl)amino/-1,3,5-triazine-2-yl)d-galacturonane with relative polymerization degree 10 and method of its preparation | |
| US4321364A (en) | Preparation of soluble chromogenic substrates | |
| CA1174234A (en) | Water-soluble cellulose mixed ethers which contain a phosphonomethyl substituent and a process for preparing same | |
| US4531000A (en) | Processes for the preparation of branched, water soluble cellulose products | |
| CS240463B1 (en) | Trisodium salt of 4-/n-ethyl-n-phenylamino/-6-/4-methyl-2-sulphobenzene-1-azo-2-/1-hydroxy-3,6-bissulphonate-1-yl/amino/-1,3,5-triazine-2-yl-hydroxyethylcellulose and method of its preparation | |
| GB900752A (en) | Polymer-dyestuff reaction products | |
| US5801239A (en) | Process for the preparation of alkali salt of carboxy alkyl cellulose | |
| RU2123500C1 (en) | Method of synthesis of n-itaconoylchitosan | |
| US4379918A (en) | Process for preparing water-soluble phosphonomethyl ethers of cellulose | |
| SK127292A3 (en) | Three-sodium salt o-{4-(n-ethyl-n-phenilamine)-6-[4-methyl-2- -sulfobenzene-1-azo-2-(1-hydroxy-3,6-bissulphonapht-1-yl)- -amino]-1,3,5-triazine-2-yl} galactomannane and method of its preparation |