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

Uses

Different sources of media describe the Uses of 3326-35-0 differently. You can refer to the following data:
1. Fluorescent labeling reagent for proteins. 4-Nitrofluorescein is used in the fluorescent antibody technique for rapid identification of pathogens.
2. Fluorescent labeling reagent for proteins. Used in the fluorescent antibody technique for rapid identification of pathogens.

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

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

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.