3326-35-0

Basic information

• Product Name: 4-NITROFLUORESCEIN
• Synonyms: 5-NITRO-3',6'-DIHYDROXYSPIRO[ISOBENZOFURAN-1(3H),9'-(9H)XANTHEN]-3-ONE;4-NITROFLUORESCEIN;4-NITROFLUOROESCEIN;3',6'-Dihydroxy-5-nitrospiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3-one;5-Nitro-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one;3',6'-dihydroxy-6-nitro-spiro[2-benzofuran-3,9'-xanthene]-1-one;3',6'-dihydroxy-6-nitrospiro[2-benzofuran-3,9'-xanthene]-1-one;3',6'-dihydroxy-6-nitro-spiro[isobenzofuran-3,9'-xanthene]-1-one
• CAS NO: 3326-35-0
• Molecular Formula: C₂₀H₁₁NO₇
• Molecular Weight: 377.3
• Product Categories: Fluorescent Labels & Indicators
• Mol File: 3326-35-0.mol
• Article Data: 4

Chemical Properties

• Melting Point: >300°C
• Boiling Point: 692°Cat760mmHg
• Flash Point: 372.3°C
• Appearance: /
• Density: 1.72g/cm³
• Vapor Pressure: 9.49E-18mmHg at 25°C
• Refractive Index: 1.764
• Storage Temp.: 2-8°C
• Solubility: almost transparency in NaOH100g/L
• PKA: 9.29±0.20(Predicted)
• Stability: Light and Moisture Sensitive
• CAS DataBase Reference: 4-NITROFLUORESCEIN(CAS DataBase Reference)
• NIST Chemistry Reference: 4-NITROFLUORESCEIN(3326-35-0)
• EPA Substance Registry System: 4-NITROFLUORESCEIN(3326-35-0)

Safety Data

• Hazardous Substances Data: 3326-35-0(Hazardous Substances Data)

Usage

Description

4-Nitrofluorescein is a synthetic organic compound that exhibits strong fluorescence properties. It is characterized by its ability to bind with proteins, making it a versatile tool in various scientific and medical applications. Its chemical structure allows for easy modification and conjugation, enhancing its utility in different fields.

Uses

Used in Biotechnology and Medical Research:
4-Nitrofluorescein is used as a fluorescent labeling reagent for proteins, enabling the visualization and tracking of specific biomolecules within complex biological systems. Its high fluorescence intensity and stability make it an ideal choice for various research applications.
Used in Pathogen Detection:
In the field of microbiology, 4-Nitrofluorescein is employed in the fluorescent antibody technique for the rapid identification of pathogens. This technique allows for the quick and accurate detection of harmful microorganisms, which is crucial for diagnosing infections and implementing appropriate treatment strategies.
Used in Analytical Chemistry:
4-Nitrofluorescein is also utilized in analytical chemistry as a sensitive and reliable indicator for various assays and tests. Its fluorescence properties make it suitable for detecting and quantifying specific substances, contributing to the advancement of chemical analysis methods.
Used in Environmental Monitoring:
In environmental science, 4-Nitrofluorescein is used as a tracer to study the behavior of pollutants and contaminants in natural systems. Its ability to be easily detected and monitored helps researchers understand the distribution and impact of harmful substances on ecosystems.

InChI:InChI=1/C20H11NO7/c22-11-2-5-15-17(8-11)27-18-9-12(23)3-6-16(18)20(15)14-4-1-10(21(25)26)7-13(14)19(24)28-20/h1-9,15,23H

Upstream product

• 5466-84-2 4-nitrophthalic anhydride 
• 108-46-3 recorcinol 
• 610-27-5 4-nitrophthalic acid 

Downstream Products

• 195977-43-6 5-nitrofluorescein dibenzoate 
• 3326-32-7 fluorescein isothiocyanate 
• 3326-34-9 fluoresceinamine 
• 153120-93-5 5-amino-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid methyl ester 

Relevant articles and documents

Synthesis and fluorescence properties of six fluorescein-nitroxide radical hybrid-compounds

Six fluorescein-nitroxide radical hybrid-compounds (2ab, 3ab, 4, and 5) were synthesized by the condensation of 5- or 6-carboxy-fluorescein and 4-amino-TEMPO (2ab), 5- or 6-aminofluorescein and 4-carboxy-TEMPO (3ab), and fluorescein and 4-carboxy-TEMPO (4), or by reaction of the 3-hydroxyl group of fluorescein with DPROXYL-3-ylmethyl methanesulfonate (5). Fluorescence intensities (around 520 nm) after reduction of the radical increased to 1.43-, 1.38-, and 1.61-folds for 2a, 2b and 3b respectively; 3a alone exhibited a decrease in intensity on reduction. Since 4 was readily solvolyzed in PBS or even methanol to afford fluorescein and 4-carboxy-TEMPO, its fluorescence change could not be measured. Hybrid compound 5 containing an ether-linkage between the fluorescein phenol and 3-hydroxymethyl-DPROXYL hydroxyl centers, was stable and on reduction, showed a maximum increase (3.21-fold) in relative fluorescence intensity in PBS (pH 5.0), despite its remarkably low absolute fluorescence intensity.

PROCESS FOR THE PREPARATION OF 5-/6-NITROFLUORESCEIN

A process for the preparation of a mixture of 3',6'-dihydroxy-6-nitrospiro[2-benzofuran-3,9'-xanthene]-1-one and 3',6'-dihydroxy-5-nitrospiro[2-benzofuran-3,9'- xanthene]-1-one comprising the steps of:- (a) reacting 4-nitrophthalic acid or 4-nitrophthalic anhydride with benzene-1,3- diol in methanesulphonic acid; (b) quenching the reaction in step (a) with a solvent to precipitate product; (c) isolating the precipitate; (d) heating the precipitate in water in order to hydrolyse any methansulphonic acid ester present.