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Chemical fluorescent molecules Chemiluminescence method Chemical properties Uses Production method Hazards & Safety Information
3-Aminophthalhydrazide structure
Chemical Name:
LUMINOL;NSC 5064;LuMinol 97%;Luminol ,98%;LuMinol AURORA KA-3798;Dia0003Luminol;Diogenes Reagent;LuminolFreeAcid99%;RARECHEM AH BS 0053
Molecular Formula:
Formula Weight:
MOL File:

3-Aminophthalhydrazide Properties

Melting point:
>300 °C(lit.)
Boiling point:
309.07°C (rough estimate)
1.3393 (rough estimate)
refractive index 
1.6500 (estimate)
storage temp. 
Solubility Insoluble in water, soluble in dimethyl sulfoxide, base
(Calcd.) 10.50 ± 0.20;;0.58 ± 0.20(at 25℃)
Dark green to dark blue-purple
PH Range
NonH uorescence (6.0) to blue H uorescence (7.0)
Water Solubility 
<0.1 g/100 mL at 19 ºC
Stable. Combustible. Incompatible with strong oxidizing agents, strong acids, strong bases, strong reducing agents. Emits light on reaction with oxidizers.
Major Application
Night vision devices, identification of product forgeries, determination of nitride in food, analytical chemistry, pharmaceuticals, iodine in biological materials, evaluating phagocytes, detecting tumors, DNA, biosensors, assay of fungi, bacteria, yeasts, assay of lipase activity
CAS DataBase Reference
521-31-3(CAS DataBase Reference)
NIST Chemistry Reference
EPA Substance Registry System
1,4-Phthalazinedione, 5-amino-2,3-dihydro-(521-31-3)
  • Risk and Safety Statements
  • Hazard and Precautionary Statements (GHS)
Hazard Codes  Xi,Xn
Risk Statements  36/37/38-22-20/21/22
Safety Statements  26-36/37-37/39-36
WGK Germany  3
RTECS  TH8890060
HS Code  29339990
Signal word: Warning
Hazard statements:
Code Hazard statements Hazard class Category Signal word Pictogram P-Codes
H302 Harmful if swallowed Acute toxicity,oral Category 4 Warning P264, P270, P301+P312, P330, P501
H315 Causes skin irritation Skin corrosion/irritation Category 2 Warning P264, P280, P302+P352, P321,P332+P313, P362
H319 Causes serious eye irritation Serious eye damage/eye irritation Category 2A Warning P264, P280, P305+P351+P338,P337+P313P
H335 May cause respiratory irritation Specific target organ toxicity, single exposure;Respiratory tract irritation Category 3 Warning
Precautionary statements:
P264 Wash hands thoroughly after handling.
P264 Wash skin thouroughly after handling.
P280 Wear protective gloves/protective clothing/eye protection/face protection.
P305+P351+P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continuerinsing.

3-Aminophthalhydrazide price More Price(16)

Manufacturer Product number Product description CAS number Packaging Price Updated Buy
Sigma-Aldrich 123072 Luminol 97% 521-31-3 2.5g $41.5 2018-11-13 Buy
Sigma-Aldrich 123072 Luminol 97% 521-31-3 5g $61.2 2018-11-13 Buy
TCI Chemical A5301 Luminol [Chemiluminescence Reagent] >98.0%(HPLC)(T) 521-31-3 1g $20 2018-11-22 Buy
TCI Chemical A5301 Luminol [Chemiluminescence Reagent] >98.0%(HPLC)(T) 521-31-3 25g $155 2018-11-22 Buy
Alfa Aesar A14597 3-Aminophthalhydrazide, 98% 521-31-3 25g $161 2018-11-13 Buy

3-Aminophthalhydrazide Chemical Properties,Uses,Production

Chemical fluorescent molecules

3-aminophthalhydrazide is a chemically fluorescent molecule. In the presence of hydrogen peroxide, it can be converted to an excited state of phthalic acid; the later one can emit a strong fluorescence. Hydrogen peroxide is the product of many biological oxidation reactions. Therefore, it is easy to introduce 3-aminophthahydrazde to connect these biological oxidation reactions with light detection. For example, the glucose oxidase/catalase probe can be used to detect the concentration of hydrogen peroxide or glucose in a sample with a response time as short as only 0.5 s (dynamic method).
Chemiluminescence immunoassay (CLIA) refers to a method of combining the chemiluminescence with immunoassay, using optical reaction to indicator the concentration of the measured immunological components. In 1976, Shmeder had proposed chemiluminescence immunoassay, using the 3-aminophthalhydrazide-H202 and its derivative ABEI as the reaction detection system. Currently, 3-aminophthalhydrazide and its derivatives have been the most commonly used markers in chemiluminescence immunoassay. They, under alkaline conditions, catalyze through microperoxidases to release large amounts of photons. 3-aminophthalhydrazide can be applied to the western blot using HRP-labeled antibody as well as the nucleic acid hybridization assays using HRP labeled antibodies. It can also be applied to the modern blood detection of criminal investigation.
A chemiluminescent material that can be used as a chemiluminescent label should meet the following requirements: the quantum yield of luminescence is high; its physicochemical properties can match the studied system; its luminescent reaction is the result of the oxidation reaction of luminescent material; when applied in the concentration range, it is biological non-toxic. Several commonly used chemiluminescent reagents include: acridinium ester is a versatile chemiluminescent tracer, being a kind of tricyclic organic compounds and easy to subject to oxidation. The oxidation reaction doesn’t need a catalyst and will release photons at the wavelength of 430nm; 3-aminophthalhydrazide and isoluminol and its derivatives are the most mature chemiluminescence agent. At as early as 1964, While had reported the study of its chemiluminescence. 3-aminophthalhydrazide, isoluminol and its derivatives have already achieved excellent results when being used in the chemiluminescence immunoassay.
Detection of luminosity: The optimum fluorescence wavelength was 400 nm (detection of chemiluminescence in the solution of 60 mM K2S2O8 and 100 mM K2CO3 with a pH of 11.5)
chemical structure of the 3-aminophthalhydrazide.
Figure 1 chemical structure of the 3-aminophthalhydrazide.

Chemiluminescence method

Chemilum inescence, as a sensitive method, has been widely used for the detection of the free radicals and reactive metabolites produced by enzymes, cells and biological organisms. The light emitted by their oxidation products and metabolites can be detected by various photometers. Because of its sensitivity, rapidity, simple operation and low cost, chemiluminescence is widely used in the screening and research of antioxidants, such as polyphenols, polysaccharides, flavonoid and anthraquinones. Commonly used chemiluminescence system for determination of superoxide anion includes xanthine oxidase-3-aminophthalhydrazide, pyrogallol-3-aminophthalhydrazide and dimethyl sulfoxide-3-aminophthalhydrazide three systems. The former belongs to the enzymatic system while the latter two belong to non-enzymatic system.
The chemiluminescent system for the determination of hydroxyl radicals mainly includes copper sulfate-yeast (or bone marrow cells)-ascorbic acid-hydrogen peroxide, CuCl-H2O2-phenanthroline-carbonate buffer, copper sulfate-phenanthroline-ascorbic acid-hydrogen peroxide , Ferrous sulfate-3-aminophthalhydrazide-hydrogen peroxide and ferrous sulfate-3-aminophthalhydrazide, five chemiluminescent systems, both involving the classical Fenton reaction to generate hydroxyl radicals, followed by attacking the luminescent agent to produce chemiluminescence, which can be used to measure the activity of the extract in removal the active oxygen free radicals.
Experimental method: apply pyrogallol-3-aminophthalhydrazide chemiluminescence system: 3-aminophthalhydrazide is formulated into a solution of 0.05mol/L with 0.05 mol/L of NaOH solution and stored in the dark place. Before usage, dilute the double distilled water to 1mmol/L solution; pyrogallol is prepared into a 0.01 mol/L solution with 1mmol/L HCl and stored at 4 ℃ refrigerator. Dilute it into a 16 times 6.25 × 10-4mol/L Solution with double distilled water before using it. The 0.05mol/L pH10.2 Na2CO3-NaHCO3 buffer (containing 0.1mmol/L EDTA) should be prepared just before usage and mixed with 1mmol/L 3-aminophthalhydrazide in a ratio of 2: 1 (volume fraction) into 3-aminophthalhydrazide and carbonate salt buffer mixture. Upon the measurement, 10.0 μL of samples (0, 0.08, 0.4, 2 and 10 mg/mL) of different concentration (with sample buffer as controls) were injected into the cell, and then injected 0.05 mL of 6.25 × 10-4 mol/L pyrogallol, and finally add 0.94 mL of the mixture of 3-aminophthalhydrazide and carbonate buffer (0.94mL) to initiate the reaction (30 ℃). The luminous intensity was recorded at 2s intervals, and a total luminescence intensity of 300 s was measured. The blank luminescence intensity refers to the luminescence value in the absence of pyrogallol.
There are also determination methods of chemiluminescence such as chemical fluorescence, fluorescence spectroscopy and NBS-dichlorofluorescein chemiluminescence system with the biggest advantage being the high sensitivity.
The above information is edited by Xiao Nan of Chemicalbook.

Chemical properties

It appears as yellow crystalline powder, being easily soluble in lye, soluble in dilute acid but almost insoluble in water and hardly soluble in alcohol. When its neutral or weak acidic solution is exposed to ultraviolet light, it exhibits strongly bright blue fluorescence. Melting point: 329-332 ° C


1.    It can be used as chemical analysis reagents, indicators.
2.    It can be used for chemiluminescence analysis such as measurement of metal cations, blood and glucocorticoids

Production method

It is obtained through the cyclization and reduction of 3-nitrophthalic acid. Add the 3-nitrophthalic acid into 8% aqueous hydrazine solution and melt the solid by heating. Add the ethylene glycol and evaporate the excess amount of water of the solution in the evaporator; heat at 215-220 °C for 2 min and close the heterocyclic ring with cooling generating yellow nitro compounds. After cooling, add 10% sodium hydroxide solution, stir and add sodium dithionite, heat to boiling and maintain the temperature for 5min, then add acetic acid with cooling obtaining bright yellow product. After standing overnight, the product was filtered.

Hazards & Safety Information

Category Toxic substances
Toxic grading poisoning
Acute toxicity Oral – rat; LD50: > 500 mg/kg
Flammability and Hazardous properties it is combustible with combustion producing toxic nitrogen oxide fumes
Storage and transportation characteristics warehouse: ventilated, low temperature and dry
Fire extinguishing agent dry powder, foam, sand, carbon dioxide, mist water

Chemical Properties

yellow crystals or beige powder


Luminol is a chemiluminescent probe that has been used to detect myeloperoxidase-mediated oxidative events in granulocytes and for chemiluminescence analysis of metal cations and blood.


Detection of copper, iron, peroxides, cyanides.

General Description

Yellow crystals or light beige powder.

Air & Water Reactions

Insoluble in water.

Reactivity Profile

Oxidation of 3-Aminophthalhydrazide is accompanied by a striking emission of light. . .

Health Hazard

ACUTE/CHRONIC HAZARDS: When heated to decomposition 3-Aminophthalhydrazide emits toxic fumes.

Fire Hazard

Flash point data for 3-Aminophthalhydrazide are not available, but 3-Aminophthalhydrazide is probably combustible.

Purification Methods

Dissolve luminol in KOH solution, treat with Norit (charcoal), filter and precipitate it with conc HCl. [Hardy et al. Talanta 24 297 1977.] Store it in the dark in an inert atmosphere, because its structure changes during its luminescence. It has been recrystallised from 0.1M KOH [Merenyi et al. J Am Chem Soc 108 77716 1986]. [Beilstein 25 II 389, 25 III/IV 4192.]

3-Aminophthalhydrazide Preparation Products And Raw materials

Raw materials

Preparation Products

3-Aminophthalhydrazide Suppliers

Global( 262)Suppliers
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Frapp's ChemicalNFTZ Co., Ltd.
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View Lastest Price from 3-Aminophthalhydrazide manufacturers

Image Release date Product Price Min. Order Purity Supply Ability Manufacturer
2018-07-25 3-Aminophthalhydrazide
US $597.00 / KG 1KG 99% 20kg career henan chemical co

3-Aminophthalhydrazide Spectrum

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