4091-99-0

Basic information

• Product Name: 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&
• Synonyms: 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&;2-[3,6-bis(acetyloxy)-2,7-dichloro-9h-xanthen-9-yl]-benzoicaci;2,7-Dichlorodihydrofluorescein diacetate (2,7-Dichlorofluorescin diacetate;2',7'-DICHLOROFLUORESCIN DIACETATE, FOR FLUORESCENCE;2',7'-Dichlorodihydrofluoresceinediacetate;Dichlorofluorescin diacetate;2'',7''-Dichlorofluorescein diac;2-(2,7-Dichloro-3,6-diacetoxy-9H-xanthen-9-yl)benzoic acid
• CAS NO: 4091-99-0
• Molecular Formula: C₂₄H₁₆Cl₂O₇
• Molecular Weight: 487.28564
• EINECS: 2017-001-1
• Mol File: 4091-99-0.mol
• Article Data: 1

Chemical Properties

• Melting Point: 209-210 °C(lit.)
• Boiling Point: 593.5 °C at 760 mmHg
• Flash Point: 312.7 °C
• Appearance: /
• Density: 1.463g/cm³
• Storage Temp.: −20°C
• Solubility: DMF: soluble
• PKA: 3.79±0.36(Predicted)
• BRN: 4610273
• CAS DataBase Reference: 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&(CAS DataBase Reference)
• NIST Chemistry Reference: 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&(4091-99-0)
• EPA Substance Registry System: 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&(4091-99-0)

Safety Data

• Hazard Codes: Xi
• Statements: 36/37/38
• Safety Statements: 22-24/25-26
• WGK Germany: 3
• F: 10-21
• Hazardous Substances Data: 4091-99-0(Hazardous Substances Data)

Usage

Description

2' 7'-DICHLOROFLUORESCIN DIACETATE FOR&, commonly known as DCFH, is a cell-permeable non-fluorescent probe used as an indicator of peroxynitrite formation and reactive oxygen species (ROS) within cells. It is supplied as the diacetate ester and, upon enzymatic or base-catalyzed cleavage of the diacetate groups, is readily oxidized to the highly fluorescent product dichlorofluorescein (DHF). The formation of DHF can be monitored by fluorescence spectroscopy using excitation and emission wavelengths of 502 and 523 nm, respectively, or by absorbance spectroscopy at 500 nm.

Uses

Used in Cellular Biology and Oxidative Stress Research:
2' 7'-DICHLOROFLUORESCIN DIACETATE FOR& is used as a cell-permeable fluorogenic probe for detecting enzymatically formed hydrogen peroxide and measuring reactive oxygen species (ROS) within cells. This application is crucial for understanding oxidative stress and its effects on cellular processes.
Used in Microplate Assays:
In this application industry, 2' 7'-DICHLOROFLUORESCIN DIACETATE FOR& is used as a sensitive and rapid quantitation tool for oxygen-reactive species in response to oxidative metabolism. It aids in detecting oxidative products in phagocytic cells, which is essential for studying immune responses and cellular defense mechanisms.
Used in Quantitative Multiwell Myeloid Differentiation Assays:
2' 7'-DICHLOROFLUORESCIN DIACETATE FOR& is employed as a fluorescent probe in quantitative multiwell assays to assess myeloid differentiation. This application helps researchers investigate the process of cell differentiation and its implications in various biological and pathological contexts.

Biochem/physiol Actions

Cell permeable: yes

Synthetic route

2',7'-dichlorodihydrofluorescein diacetate (2044-85-1) → 2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0)

Conditions	Yield
With palladium 10% on activated carbon; hydrogen In ethyl acetate at 20℃; under 760.051 Torr; for 18h;

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) + propargyl alcohol (107-19-7)

Conditions	Yield
With dmap; dicyclohexyl-carbodiimide In dichloromethane at 20℃; for 1h;	61%

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → 2,7-dichlorofluorescein

Conditions	Yield
With dihydrogen peroxide; horseradish peroxidase In water
Multi-step reaction with 2 steps 1: sodium hydroxide / ethanol / 0.5 h / 20 °C 2: 3-(N-morpholino)propanesulfonic acid; cysteamine; air / CuSO4; 1,10-phenanthroline / 25 °C / pH 7 View Scheme
With Ac-IHIHIQI-CONH2; water; copper(II) ion pH=8; Kinetics; Reagent/catalyst;
Multi-step reaction with 2 steps 1: sodium hydroxide / ethanol 2: C35H29CuN3O4 / ethanol; aq. phosphate buffer / pH 7.4 View Scheme

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → 2-(2,7-dichloro-3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid (106070-31-9)

Conditions	Yield
With sodium hydroxide In ethanol at 20℃; for 0.5h;
With sodium hydroxide for 0.5h; cooling;
With sodium hydroxide In ethanol at 20℃; for 0.5h; Darkness;

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → 2',7'-dichlorofluorescein (76-54-0)

Conditions	Yield
With hemin; d(ATTGTGCGTCATCCCTTACGTCAGTGGAG), screened from Cauliflower mosaic virus (CaMV) 35S promoter; d(CTCCACTGACGTAATGGGTAGGG); d(GGGTTGGGCGGATGACGCACAAT); potassium chloride; ammonium acetate In water at 20℃; for 0.166667h; pH=7; aq. buffer;
With water; oxygen Irradiation;

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C29H22Cl2O7

Conditions	Yield
Multi-step reaction with 2 steps 1: dicyclohexyl-carbodiimide; dmap / dichloromethane / 18 h / 20 °C / Inert atmosphere 2: ammonium bicarbonate / tetrahydrofuran; methanol; water / 18 h / 20 °C / Inert atmosphere; Darkness View Scheme

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C35H30Cl2O7

Conditions	Yield
Multi-step reaction with 3 steps 1: dicyclohexyl-carbodiimide; dmap / dichloromethane / 18 h / 20 °C / Inert atmosphere 2: ammonium bicarbonate / tetrahydrofuran; methanol; water / 18 h / 20 °C / Inert atmosphere; Darkness 3: tetra(n-butyl)ammonium hydrogensulfate; potassium carbonate / dichloromethane; water / 18 h / 20 °C / Inert atmosphere; Darkness View Scheme

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C26H18Cl2O5

Conditions

Yield

Multi-step reaction with 4 steps 1: dicyclohexyl-carbodiimide; dmap / dichloromethane / 18 h / 20 °C / Inert atmosphere 2: ammonium bicarbonate / tetrahydrofuran; methanol; water / 18 h / 20 °C / Inert atmosphere; Darkness 3: tetra(n-butyl)ammonium hydrogensulfate; potassium carbonate / dichloromethane; water / 18 h / 20 °C / Inert atmosphere; Darkness 4: 2,3-dicyano-5,6-dichloro-p-benzoquinone / dichloromethane; water / 18 h / 20 °C / Inert atmosphere; Darkness View Scheme

2,4-dimethoxylbenzyl alcohol (7314-44-5) + 2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C33H26Cl2O9

Conditions	Yield
With dmap; dicyclohexyl-carbodiimide In dichloromethane at 20℃; for 18h; Inert atmosphere;	149.4 mg

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C50H45Cl3N6O12S

Conditions	Yield
Multi-step reaction with 2 steps 1: dicyclohexyl-carbodiimide; dmap / dichloromethane / 1 h / 20 °C 2: copper(II) sulfate; tris[(1-benzyl-1H-1,2,3-triazol-4yl)methyl]amine; sodium L-ascorbate / tetrahydrofuran; water; ethanol / 2 h / 20 °C View Scheme

2-(3,6-diacetyloxy-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (4091-99-0) → C46H41Cl3N6O10S

Conditions	Yield
Multi-step reaction with 3 steps 1: dicyclohexyl-carbodiimide; dmap / dichloromethane / 1 h / 20 °C 2: copper(II) sulfate; tris[(1-benzyl-1H-1,2,3-triazol-4yl)methyl]amine; sodium L-ascorbate / tetrahydrofuran; water; ethanol / 2 h / 20 °C 3: sodium hydroxide / methanol / 0.5 h / 37 °C View Scheme

Upstream product

• 2044-85-1 2',7'-dichlorodihydrofluorescein diacetate 

Downstream Products

• 76-54-0 2',7'-dichlorofluorescein 
• 106070-31-9 2-(2,7-dichloro-3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid 
• 18362-80-6 2,7-dichlorofluorescein 

Relevant articles and documents

Synthesis of high contrast fluorescein-diethers for rapid bench-top sensing of palladium

A new series of palladium sensors based on fluorescein bis(allyl ether) compounds has been designed. We utilized reduced fluoresceins as key synthetic intermediates. These probes exhibit negligible background fluorescence and rapid reaction with palladium, allowing a concentration of 100 ppt to be detected in minutes using a handheld UV lamp. This journal is