CS229094B1 - Trisodium salt of 6-/2-sulpho-4/4-amino-3-sulpho-1-anthraquinoneamino/anilino-4-/x-sulphoanilino/-1,3,5-triazin-2-yl-hydroxyethyl starch and method of preparing same - Google Patents

Trisodium salt of 6-/2-sulpho-4/4-amino-3-sulpho-1-anthraquinoneamino/anilino-4-/x-sulphoanilino/-1,3,5-triazin-2-yl-hydroxyethyl starch and method of preparing same Download PDF

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CS229094B1
CS229094B1 CS39183A CS39183A CS229094B1 CS 229094 B1 CS229094 B1 CS 229094B1 CS 39183 A CS39183 A CS 39183A CS 39183 A CS39183 A CS 39183A CS 229094 B1 CS229094 B1 CS 229094B1
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sulfo
blue
amino
anilino
hydroxyethyl starch
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CS39183A
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Slovak (sk)
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Peter Gemeiner
Danica Mislovicova
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Peter Gemeiner
Danica Mislovicova
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Description

Vynález sa týká trojsodnej soli 6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino )J anilino-4-(x-sulfoaniIino)-1,3,5-triazín-2-yl-hydroxyetylškrobu a spfisobu jeho přípravy.The present invention relates to the trisodium salt of 6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine-2- yl-hydroxyethyl starch and methods for its preparation.

Trojsodná sol’ 6- [2-sulf o-4-( 4-amino-3-sulfo-1 -antrachinón-amino)] anilino-4-(x-sulfoanilino)-! ,3,5--triazín-2-yl-dextránu je vodorozpustná látka a slúžila v afinitnej chromato grafii ako prostriedok na štúdium afinity enzýmov a proteínov vfibeo k antrachinón-triazínovým farbivám,, ktoré sú jeuným z typov generálnych afinantov (E. Stellwagen: Accounts Chem. Res. 10, 92 /1977/)· Chromolytioké dextrány o priemernej molekulovej hmotnosti 5.10^ až 2.10^ sa vyznačovali výraznou afinitou k enzýmom, ktorá je závislá nielen od typu chromofóru, ale aj od molekulovej hmotnosti polysacharidu. Zo skúšaných antrachlnón-triazínových farbív najvačšiu účinnost vykazovala trojsodná sol’ 2-chlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antraehinón-amino)]anilino-4-(x-sulfoanilino)-1,3,5-triazínu (P. Gemeiner,Trisodium salt '6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -! 3,5-triazin-2-yl-dextran is a water-soluble substance and has been used in affinity chromatography as a means to study the affinity of vfibeo enzymes and proteins for anthraquinone-triazine dyes, which are a type of general affinity (E. Stellwagen: 10, 92 (1977)) Chromolytic dextrans with an average molecular weight of 5.10 to 2.10 were characterized by a strong affinity for enzymes, which is dependent not only on the type of chromophore but also on the molecular weight of the polysaccharide. Of the anthraquinone-triazine dyes tested, the trisodium salt of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) showed the greatest activity -1,3,5-triazine (P. Gemeiner,

D. Mislovičová, J. Zemek, t. Kuniak: Collect. Czeoh. Chem. Commun. 46, 419 /1981/). Vplyv charakteru polysacharidu na afinitu jeho chromolytického derivátu ku enzýmom a albuminu bol zatial’ dokázaný z rozpustných polysaoharidov iba u dextránu, rozpustného škrobu a glykogénu a z nerozpustných polysaoharidov u sieťovaných dextránov, sieťovanej agarózy a celu lózy (S. Angal, P. D. G. Dean: Biochem. J. 167, 301 /1977/).D. Mislovicova, J. Zemek, t. Kuniak: Collect. Czeoh. Chem. Commun. 46, 419 (1981)]. The effect of polysaccharide character on the affinity of its chromolytic derivative for enzymes and albumin has so far been demonstrated from soluble polysaoharides only in dextran, soluble starch and glycogen, and insoluble polysaoharides in crosslinked dextrans, cross-linked agarose and cellulose (S. Angal, PDG Dean. 167, 301 (1977)].

Hydroxyetylškrob je ňalší typ vodorozpustríého polysacharidu vhodného na tieto účely. Je to vetvený alfa-glukán (99 % amylopektín) vo formě jeho hydroxyetylderivátu so stupňom substitúcie 0,63 až 0,67. Frakcionáciou tohto vodorozpustného polysacharidu sa získali frakcie hydroxyetylškrobu s priemernou molekulovou hmotnosťou v rozmedzl 5.10^ až 2.10^. Pri vypracovaní podmienok reakcie hydroxyetylškrobu s trojsodnou sol’ou 2-chlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazínom sa vychádzalo z podmienok farbenia dextránov (P. Gemeiner a kol.: Collect. Czech. Chem. Com229094 mun., 46, 4)9 /1981/). Aby sa dosiahol jednotný stupeň substitdeie dextránov s trójaodnou soTou 2-chlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)J anilino-4-Cx-sulfoanilino)-1,3,5-triazínu (65 umól/g), používali sa hmotnostně poměry trojsodná sol’ 2-chlór-6- 2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino) anilino-4-(x-sulfoanilino)-1,3,5-triazínu : dextrán : uhličitén sodný = 0,5:1:2 s tým, že reakcia prebiehala po dobu 2 h pri teplote 80 °G. Za týchto podmienok sa viazalo len 10 % trojsodnej soli 2-chlór-6- 2-sulfo-4-(4-amino-3-sulfo-1 -antrachinón-amino) anilino-4-(x-sulf'oanilino)-1,3,5-triazínu, So je ekonomicky nákladné.Hydroxyethyl starch is another type of water-soluble polysaccharide suitable for this purpose. It is a branched alpha-glucan (99% amylopectin) in the form of its hydroxyethyl derivative with a degree of substitution of 0.63 to 0.67. Fractionation of this water-soluble polysaccharide yields hydroxyethyl starch fractions having an average molecular weight in the range of 5.10 to 2.10. In working out the reaction conditions of hydroxyethyl starch with trisodium salt of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1 3,5-triazine was based on dextran staining conditions (P. Gemeiner et al .: Collect. Czech. Chem. Com229094 mun., 46, 4 (9), 1981). In order to achieve a uniform degree of substitution of the dextrans with the triiodium salt of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4-Cx-sulfoanilino) -1, Of 3,5-triazine (65 µmol / g), the weight ratios of the trisodium salt of 2-chloro-6- 2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino) anilino-4- (x-sulfoanilino) -1,3,5-triazine: dextran: sodium carbonate = 0.5: 1: 2 with reaction at 80 ° C for 2 h. Under these conditions, only 10% of the trisodium salt of 2-chloro-6- 2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino) anilino-4- (x-sulfoanilino) -1 bound 3,5-triazine, SO is economically expensive.

Uvedenú nevýhodu odstraňuje v podstatnej miere tento vynález. Podstatou vynálezu je trojsodná sol’ 6-[^2-sulfo-4-(4-amino-3-sulfo-1-antraohinón-amino)J anilino-4-(x-sulfoanilino)-1,3,5-triazín-2-yl-hydroxyetylěkrobu, kde x je poloha substituenta 2 alebo 4 vzorca IThis disadvantage is substantially eliminated by the present invention. The present invention provides the trisodium salt of 6 - [[2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine- 2-yl-hydroxyethyl starch, wherein x is the position of substituent 2 or 4 of formula I

kde R představuje hydroxyetylSkrob so stupňom substitúcie 0,63 až 0,67 a o priemernej molekulovej hmotnosti 5.10^ až 2.10^. Dalej podstatou vynálezu je spOsob přípravy týchto derivátov, ktorý sa vyznačuje tým, že jednotlivé frakcie hydroxyetylěkrobu sa po dobu 30 min aktivujú vodným roztokom trojsodnej soli 2-chlór-6-[2-suIfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazínu vzorca II pri teplote 60 °C a po přidaní chloridu sodného a uhličitanu sodného reaguji! s trojsodnou soTou 2-chlór-6- [2-sulfo -4-(4-amino-3-sulfo-1-antrachinón-amino)J anilino-4-(x-sulfoanilino)-1,3,5-triazínu po dobu 2 h pri teplote 70 °C při hmotnostných poraeroch trojsodná sol’ 2-ohlór-6-[2-sulfo-4-(4-amino -3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazínu : hydroxyetylSkrob : uhličitan sodný = 0,18 až 0,25:1:0,2 v prostředí 3,5 % vodného roztoku chloridu sodného.wherein R represents a hydroxyethyl starch with a degree of substitution of 0.63 to 0.67 and an average molecular weight of 5.10 to 2.10. Another object of the present invention is a process for the preparation of these derivatives, characterized in that the individual fractions of hydroxyethyl starch are activated for 30 minutes with an aqueous solution of the trisodium salt of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo). 1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine of the formula II at 60 ° C and after addition of sodium chloride and sodium carbonate, reacts! with trisodium salt of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x- sulfoanilino) -1,3,5-triazine: hydroxyethylstarch: sodium carbonate = 0.18 to 0.25: 1: 0.2 in 3.5% aqueous sodium chloride solution.

(II)(II)

Výhodou navrhovaného spOsobu přípravy oproti doterajěiemu spOsobu přípravy chromolytických dextránov je, že pri 2 až 5násobne nižěej spotrebe ti>ojsodnej soli 2-ohlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazínu, lOnásobne nižgej spotrebe uhličitanu sodného a pri znížení teploty o 10 °C sa dosahuje rovnaký stupeň substitúcie a)co pri farbení dextránov a že uvedený postup je jednoduchý a nenáročný na použité chemikálie a přístroje.The advantage of the proposed preparation method over the prior art preparation of chromolytic dextrans is that at 2 to 5 times lower consumption of the 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino) thiodium salt. )] anilino-4- (x-sulfoanilino) -1,3,5-triazine, 10 times lower consumption of sodium carbonate and lowering temperature by 10 ° C achieves the same degree of substitution a) as in dextran dyeing and that the procedure is simple and easy on used chemicals and instruments.

Příklad 1 mg hydroxyetylškrobu (HES) o priemernej molekulovej hmotnosti 53 500 a 4,9 mg trojsodnej soli 2-chlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-aminoj] anilino-4-(x-sulfoanilino)-1,3,5-triazínu (Cibacron Blue 3G-A Sálej Blue) sa rozpustí v ,,25 ml destiiovanej vody při teplote 60 °C a mieša sa po dobu 30 min. Potom sa přidé 43,75 mg chloridu sodného a zmes sa zohreje na teplotu 70 °G. Po přidaní 5 mg uhličitanu sodného sa reakčná zmes mieša ešte po dobu 2 h. Potom sa reakčná zmes ochladí, zneutralizuje zriedenou kyselinou chlorovodíkovou na pH 7 a přečistí na koloně naplnenej dextránom sieťovaným epichlórhydrínom (Sephadex G-25 fine). Objem náplně kolony je 10C ml, výška 21,5 cm a priemer 2,5 cm. Eluuje sa destilovanou vodou, pri prietoku eluátu kolonou 30 až 35 ml/h.Example 1 mg of hydroxyethyl starch (HES) with an average molecular weight of 53,500 and 4.9 mg of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino- trisodium salt Dissolve 4- (x-sulfoanilino) -1,3,5-triazine (Cibacron Blue 3G-A Sale Blue) in 25 mL of distilled water at 60 ° C and stir for 30 min. 75 mg of sodium chloride and heated to 70 [deg.] C. After the addition of 5 mg of sodium carbonate, the reaction mixture was stirred for 2 h. The reaction mixture was cooled, neutralized with dilute hydrochloric acid to pH 7 and purified on a dextran column. The column volume is 10 ml, height 21.5 cm and diameter 2.5 cm, eluted with distilled water at a flow rate of 30 to 35 ml / h through the column.

Prvá modrá frakcia obsahujdca vodný roztok trojsodnej soli 6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazín-2-yl-hydroxyetylškrobu (Cibacron Blue-HES, ďalej Blue-HES) sa odpaří na rotačnej vákuovej odparke do sucha pri tlaku 5 kPa; po přidaní a znovu odpaření etanolu a potom acetonu sa získá suchý Blue-HES (23,1 mg) s obsahom 65,4 umólu Blue/g.The first blue fraction containing an aqueous solution of the trisodium salt of 6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine -2-yl-hydroxyethyl starch (Cibacron Blue-HES; hereinafter Blue-HES) is evaporated to dryness on a rotary evaporator at 5 kPa; after addition and re-evaporation of ethanol and then acetone, dry Blue-HES (23.1 mg) was obtained containing 65.4 µmol Blue / g.

Příklad 2Example 2

Postup podTa příkladu 1 s tým rozdielom, že sa použije hydroxyetylškrob o priemernej molekulovej hmotnosti ,27 000 a 4,63 mg Cibacron Blue 3G-A. Získá sa Blue-HES (22,5 mg) o obsahu 65,4 umól Blue/g.The procedure of Example 1 except that hydroxyethyl starch with an average molecular weight of 27,000 and 4.63 mg of Cibacron Blue 3G-A was used. Blue-HES (22.5 mg) was obtained with a content of 65.4 µmol Blue / g.

Příklad 3Example 3

Postup podTa příkladu 1 s tým rozdielom, že sa použije hydroxyetylškrob o priemernej molekulovej hmotnosti 402 000 a 6,23 mg Blue. Suchý Blue-HES (22,2 mg) obsahuje 68,7 Umól Blue/g.The procedure of Example 1 was followed except that hydroxyethyl starch having an average molecular weight of 402,000 and 6.23 mg of Blue was used. Dry Blue-HES (22.2 mg) contains 68.7 µmol Blue / g.

Příklad 4Example 4

Postup podTa příkladu 1 s tým rozdielom, že sa použil hydroxyetylškrob o priemernej molekulovej hmotnosti 1 920 000 a 6,23 mg Blue. Suchý Blue-HES (21,7 mg) obsahuje 72,08 umól Blue/g.The procedure of Example 1 except that hydroxyethyl starch with an average molecular weight of 1,920,000 and 6.23 mg of Blue was used. Dry Blue-HES (21.7 mg) contains 72.08 µmol Blue / g.

Vynález má použitie v enzýmovom inžinierstve a v biochémii, a to pri izoláeii a purifikácii enzýmov alebo ako substráty a inhibitory enzýmami katalyzovaných reakeií.The invention has utility in enzyme engineering and in biochemistry, either in the isolation and purification of enzymes, or as substrates and inhibitors of enzyme catalyzed reactions.

Claims (2)

3 229094 Příklad 1 25 mg hydroxyetylškrobu (HES) o priemernej molekulovej hmotnosti 53 500 a 4,9 mg troj-sodnej soli 2-chlór-6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-aminoj] anilino-4-(x-sulfo-anilino)-1,3,5-triazínu (Cibacron Blue 3G-A áalej Blue) sa rozpustí v 1,25 ml destilovanejvody při teplote 60 °C a mieša sa po dobu 30 min. Potom sa pridé 43,75 mg chloridu sodnéhoa zmes sa zohreje na teplotu 70 °G. Po přidaní 5 mg uhliíitanu sodného sa reakSná zmesmieša ešte po dobu 2 h. Potom sa reakčná zmes ochladí, zneutralizuje zriedenou kyselinouchlorovodíkovou na pH 7 a přečistí na koloně naplnenej dextránom sletovaným epichlórhydrí-nom (Sephadex G-25 fine). Objem náplně kolony je 10C ml, výška 21,5 cm a priemer 2,5 cm.Eluuje sa destilovanou vodou, prl prietoku eluátu kolonou 30 až 35 ml/h. Prvá modrá frakcia obsahujdca vodný roztok trojsodnej soli 6-j2-sulfo-4-(4-aniino-3--sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazín-2-yl-hydroxyetyl-škrobu (Cibacron Blue-HES, dalej Blue-HES) sa odpaří na rotačnej vákuovej odparke do suchapri tlaku 5 kPa; po přidaní a znovu odpaření etanolu a potom acetonu sa získá suchý Blue--HES (23,1 mg) s obsahom 65,4 umólu Blue/g. Příklad 2 Postup podl’a příkladu 1 s tým rozdielom, že sa použije hydroxyetylškrob o priemernejmolekulovej hmotnosti 127 000 a 4,63 mg Cibacron Blue 3G-A. Získá sa Blue-HES (22,5 mg)o obsahu 65,4 umól Blue/g. Příklad 3 Postup pódia příkladu 1 s tým rozdielom, Se sa použije hydroxyetylškrob o priemernejmolekulovej hmotnosti 402 000 a 6,23 mg Blue. Suchý Blue-HES (22,2 mg) obsahuje 68,7 UmólBlue/g. Příklad 4 Postup pódia příkladu 1 s tým rozdielom, že sa použil hydroxyetylškrob o priemernejmolekulovej hmotnosti 1 920 000 a 6,23 mg Blue. Suchý Blue-HES (21,7 mg) obsahuje 72,08umól Blue/g. Vynález má použitie v enzýmovom inžinierstve a v biochémii, a to pri izoláeii a purifi-kácii enzýmov alebo ako substráty a inhibitory enzýmami katalyzovaných reakcií. P K E D Μ E T VYNÁLEZU229094 Example 1 25 mg hydroxyethyl starch (HES) with an average molecular weight of 53,500 and 4.9 mg of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone) tri-sodium salt aminamine anilino-4- (x-sulfo-anilino) -1,3,5-triazine (Cibacron Blue 3G-A further Blue) is dissolved in 1.25 ml distilled water at 60 ° C and stirred for 30 minutes. Sodium chloride (43.75 mg) was added and the mixture was heated to 70 DEG C. After addition of 5 mg of sodium carbonate, the reaction mixture was stirred for 2 h. The reaction mixture was cooled, neutralized with dilute hydrochloric acid to pH 7 and purified to give a yellow oil. Column packed with dextran-milled epichlorohydrin (Sephadex G-25 fine) The column loading volume is 10C ml, 21.5 cm high and 2.5 cm in diameter.It is eluted with distilled water, the eluate flow rate is 30-35 ml / h. First blue fraction containing an aqueous solution of 6-β-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine trisodium salt -2 -yl-hydroxyethyl starch (Cibacron Blue-HES, further Blue-HES) is evaporated in a rotary evaporator to a pressure of 5 kPa; ethanol was added and re-evaporated, followed by acetone to give dry Blue-HES (23.1 mg) containing 65.4 µmol Blue / g. Example 2 The procedure of Example 1 except that hydroxyethyl starch of an average molecular weight of 127,000 and 4.63 mg of Cibacron Blue 3G-A was used. Blue-HES (22.5 mg) was obtained with a content of 65.4 µmol Blue / g. EXAMPLE 3 The procedure of Example 1, with the difference, is hydroxyethyl starch having an average molecular weight of 402,000 and 6.23 mg Blue. Dry Blue-HES (22.2 mg) contains 68.7 µmol Blue / g. EXAMPLE 4 Procedure of Example 1 except that hydroxyethyl starch having an average molecular weight of 1,920,000 and 6.23 mg Blue was used. Dry Blue-HES (21.7 mg) contains 72.08 µmol Blue / g. The invention has utility in enzyme engineering and in biochemistry for the isolation and purification of enzymes or as substrates and inhibitors of enzyme-catalyzed reactions. P K E D E T I T OF THE INVENTION 1, Trojsodná sol’ 6-[2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)J anilino-4-(x-sulfoanilino)-!,3,5-triazín-2-yl-hydroxyetylškrobu, kde x je poloha substituenta 2 alebo 4vzorca I1, 6- [2-Sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] - anilino-4- (x-sulfoanilino) -1,3,5-triazine-2- yyloxyethyl starch wherein x is the position of substituent 2 or 4 of formula I SO3Na 229094 4 kde R představuje hydroxyetylSkrob so stupňom substitúcie 0,63 až 0,67 a priemernou mole-kulovou hmotnosťou 5.10^ až 2.10®.Wherein R is hydroxyethyl starch with a degree of substitution of 0.63 to 0.67 and an average molecular weight of 5.10 to 2.10 ®. 2. Spfisob přípravy podl’a bodu 1 vyznačujúci sa tým, že frakcionovaný vodorozpustnýhydroxyetylSkrob sa po dobu 30 min aktivuje vodným roztokom trojsodnej soli 2-chlór-6-[2--sulfO”4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)-1,3,5-triazínuvzorca II2. Preparation according to claim 1, characterized in that the fractionated water-soluble hydroxyethyl starch is activated for 30 min with an aqueous solution of 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1) tri-sodium salt. -antraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine compound II při teplote 60 °C a po přidaní chloridu sodného a uhličitanu sodného reaguje s trojsodnousol’ou 2-chlór-6- [2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfo-anilino)-1,3,5-triazínu po dobu 2 h pri teplote 70 °C pri hmotnostných pomeroch trojsodnáso! 2-chlór-6-^2-sulfo-4-(4-amino-3-sulfo-1-antrachinón-amino)] anilino-4-(x-sulfoanilino)--1,3,5-triazínu : hydroxyetylSkrob : uhličitan sodný = 0,18 až 0,25:1:0,2 v prostředí 3,5 %vodného chloridu sodného. Severografia, n. p., MOST Cena 2,40 Kčsat 60 ° C, and after addition of sodium chloride and sodium carbonate, it reacts with trisodium 2-chloro-6- [2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4 - (x-sulfo-anilino) -1,3,5-triazine for 2 h at 70 ° C with a tri-sodium ratio by weight! 2-Chloro-6- (2-sulfo-4- (4-amino-3-sulfo-1-anthraquinone-amino)] anilino-4- (x-sulfoanilino) -1,3,5-triazine: hydroxyethyl sodium carbonate = 0.18 to 0.25: 1: 0.2 in 3.5% aqueous sodium chloride. Severografia, n. P., MOST Price 2,40 Kcs
CS39183A 1983-01-20 1983-01-20 Trisodium salt of 6-/2-sulpho-4/4-amino-3-sulpho-1-anthraquinoneamino/anilino-4-/x-sulphoanilino/-1,3,5-triazin-2-yl-hydroxyethyl starch and method of preparing same CS229094B1 (en)

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CS39183A CS229094B1 (en) 1983-01-20 1983-01-20 Trisodium salt of 6-/2-sulpho-4/4-amino-3-sulpho-1-anthraquinoneamino/anilino-4-/x-sulphoanilino/-1,3,5-triazin-2-yl-hydroxyethyl starch and method of preparing same

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CS39183A CS229094B1 (en) 1983-01-20 1983-01-20 Trisodium salt of 6-/2-sulpho-4/4-amino-3-sulpho-1-anthraquinoneamino/anilino-4-/x-sulphoanilino/-1,3,5-triazin-2-yl-hydroxyethyl starch and method of preparing same

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