# US 11,112,366 B2 · Reagent and kit for enhancing chemiluminescent reaction

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- Number: US 11,112,366 B2
- Document ID: US11112366B2
- Status: Granted
- Kind: B2
- Application number: 16/368,108
- Filed: 2019-03-28
- Published: 2020-10-01
- Granted: 2021-09-07
- Earliest claimed: 2019-03-28
- Assignees: National Taiwan Ocean University
- Applicants: National Taiwan Ocean University
- Inventors: Chih-Ching Huang; Chia-Wen Lien
- Web page: https://patentgrep.com/patent/US11112366B2

## CPC

- G01N 21/76: Chemiluminescence; Bioluminescence
- C07D 473/00: Heterocyclic compounds containing purine ring systems
- C07D 473/40: with halogen atoms or perhalogeno-alkyl radicals directly attached in position 2 or 6
- C07H 19/04: Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C12Q 1/28: involving peroxidase
- G01N 33/52: Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)}
- G01N 33/582: {with fluorescent label}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

A reagent for enhancing a chemiluminescent reaction includes luminol or luminol derivatives, an oxidant, an electron mediator, and an enhancer. The enhancer is a nitrogen-containing fused heterocyclic compound having at least two nitrogen atoms. The present disclosure further provides a kit for enhancing a chemiluminescent reaction comprising the foregoing reagent.

## Claims

### Claim 1

A reagent for enhancing a chemiluminescent reaction, comprising:
luminol or luminol derivatives;
an oxidant;
an electron mediator; and
an enhancer, which is a nitrogen-containing fused heterocyclic compound having at least two nitrogen atoms and is a compound selected from the group consisting of:

### Claim 2 (depends on 1)

The reagent according to claim 1, wherein the oxidant is sodium perborate or hydrogen peroxide, and the electron mediator is p-iodophenol, p-iodophenylboronic acid, sodium 3-(phenothiazin-10-yl) propane-1-sulfonate, or sodium 4-(phenothiazin-10-yl) butane-1-sulfonate.

### Claim 3 (depends on 2)

The reagent according to claim 2, wherein the oxidant is sodium perborate, and the electron mediator is sodium 3-(phenothiazin-10-yl) propane-1-sulfonate.

### Claim 4 (depends on 1)

The reagent according to claim 1, wherein the enhancer is a compound selected from the group consisting of:

### Claim 5 (depends on 1)

The reagent according to claim 1, wherein the enhancer is:

### Claim 6 (depends on 1)

The reagent according to claim 1, wherein the reagent has a pH ranging from 7.0 to 10.0.

### Claim 7 (depends on 6)

The reagent according to claim 6, wherein the reagent has a pH ranging from 7.5 to 9.0.

### Claim 8 (depends on 1)

A kit for enhancing a chemiluminescent reaction, comprising a first vial, a second vial, and the reagent according to claim 1, wherein the luminol or luminol derivatives is present in the first vial, the oxidant is present in the second vial, and the electron mediator and the enhancer are present either in the first vial or in the second vial or in both vials.

### Claim 9 (depends on 8)

The kit according to claim 8, further comprising a peroxidase.

### Claim 10 (depends on 9)

The kit according to claim 9, wherein the peroxidase is horseradish peroxidase, soybean peroxidase, or sweet potato peroxidase.

### Claim 11 (depends on 8)

The kit according to claim 8, wherein the oxidant of the reagent is sodium perborate or hydrogen peroxide, and the electron mediator of the reagent is p-iodophenol, p-iodophenylboronic acid, sodium 3-(phenothiazin-10-yl) propane-1-sulfonate, or sodium 4-(phenothiazin-10-yl) butane-1-sulfonate.

### Claim 12 (depends on 8)

The kit according to claim 8, wherein the oxidant of the reagent is sodium perborate, and the electron mediator of the reagent is sodium 3-(phenothiazin-10-yl) propane-1-sulfonate.

### Claim 13 (depends on 8)

The kit according to claim 8, wherein the enhancer of the reagent is a compound selected from the group consisting of:

### Claim 14 (depends on 13)

The kit according to claim 13, wherein the enhancer of the reagent is

### Claim 15 (depends on 8)

The kit according to claim 8, wherein the reagent has a pH ranging from 7.0 to 10.0.

### Claim 16 (depends on 15)

The kit according to claim 15, wherein the reagent has a pH ranging from 7.5 to 9.0.