Method of quantitative determination of substances using coumarin derivatives

A method of quantitatively determining one of a peroxidation-active substance, hydrogen peroxide or a coumarin derivative represented by the formula:

• ##STR30## in which R.sup.1 represents hydrogen, halogen, C.sub.1-8 linear or branched alkoxy, substituted or unsubstituted C.sub.1-8 linear or branched alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, cyano, carboxyl, C.sub.1-8 linear or branched alkoxycarbonyl, C.sub.1-8 linear or branched alkanoyl, pyridyl or benzothiazolinyl;
• R.sup.2, R.sup.3, R.sup.4 and R.sup.6 are independently hydrogen, hydroxy or substituted or unsubstituted C.sub.1-8 linear or branched alkyl;
• and
• R.sup.5 represents hydrogen, hydroxy, C.sub.1-8 linear or branched alkoxy, nitro, substituted or unsubstituted C.sub.1-8 linear or branched alkyl, substituted or unsubstituted amino, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, cyano, carboxyl, C.sub.1-8 linear or branched alkoxycarbonyl, C.sub.1-8 linear or branched alkanoyl, pyridyl or benzothiazolinyl;
• or R.sup.1 and R.sup.2 combined together form C.sub.2-4 alkylene or C.sub.2-4 alkylene or C.sub.2-4 alkenylene;
• or R.sup.4 and R.sup.5 combined together form --CH.sub.2 CH.sub.2 CH.sub.2 NH--;
• the substituted phenyl, naphthyl and alkyl being independently substituted with 1-5 cyano, halogen, amino independently substituted with 0 to 2 C.sub.1-8 linear or branched alkyl, substituted or unsubstituted heterocyclic group (wherein the heterocyclic group is independently triazinyl, pyrazinyl, pyridyl or pyrimidinyl independently substituted with 0 to 2 cyano, halogen, amino, alkoxy, hydroxy or C.sub.1-8 linear or branched dialkylamino) , C.sub.1-8 linear or branched alkyl, C.sub.1-8 linear or branched alkoxy, carboxy, C.sub.1-8 linear or branched alkoxycarbonyl and hydroxy and the substituted amino being independently substituted with 1 or 2 C.sub.1-8 linear or branched alkyl, triazinyl, pyrazinyl, pyridyl or pyrimidinyl, with the proviso that when R.sup.6 is hydrogen, at least one of R.sup.1, R.sup.2 and R.sup.3 cannot be hydrogen;
• in a sample, comprising the steps of:
• (A) selecting a known amount of two substances selected from the peroxidation-active substance, hydrogen peroxide and coumarin derivative or salt thereof;
• (B) reacting said two substances with said sample having an unknown amount of the remaining substance from said peroxidation active substance, hydrogen peroxide, or coumarin derivative or salt thereof;
• (C) determining the intensity of light emitted from the reacted solution;
• and
• (D) comparing the intensity of said emitted light with a calibration curve.

A method of quantitatively determining one of a peroxidation-active substance, hydrogen peroxide or a coumarin derivative represented by the formula:

• ##STR43## wherein R.sup.1 represents hydrogen, C.sub.1-8 linear or branched alkoxy, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, cyano, carboxyl, C.sub.1-8 linear or branched alkoxycarbonyl, C.sub.1-8 linear or branched alkanoyl, pyridyl or benzothiazolinyl;
• and R.sup.2 and R.sup.3 are independently hydrogen or substituted or unsubstituted C.sub.1-8 linear or branched alkyl;
• or R.sup.1 and R.sup.2 combined together form C.sub.2-4 alkylene or C.sub.2-4 alkylene;
• the substituted phenyl, naphthyl and alkyl are independently substituted with 1-5 cyano, halogen, amino independently substituted with 0 to 2 C.sub.1-8 linear or branched alkyl, substituted or unsubstituted heterocyclic group (wherein the heterocyclic group is independently triazinyl, pyrazinyl, pyridyl or pyrimidinyl independently substituted with 0 to 2 cyano, halogen, amino, alkoxy, hydroxy or C.sub.1-8 linear or branched dialkylamino), C.sub.1-8 linear or branched alkyl, C.sub.1-8 linear or branched alkoxy, carboxy, C.sub.1-8 linear or branched alkoxycarbonyl and hydroxy;
• in a sample, comprising the steps of:
• (A) selecting a sample having a known amount of two substances selected from the peroxidation-active substance, hydrogen peroxide and coumarin derivative or salt thereof;
• (B) reacting said two substances with said sample having an unknown amount of the remaining substance from said peroxidation active substance, hydrogen peroxide, or coumarin derivative or salt thereof;
• (C) determining the intensity of light emitted from the reacted solution;
• and
• (D) comparing the intensity of said emitted light with a calibration curve.