5-Bromo-4-chloro-3-indoxyl alpha-D-glucopyranoside
[CAS# 108789-36-2]

Identification

• Classification: Biochemical >> Carbohydrate >> Monosaccharide
• Name: 5-Bromo-4-chloro-3-indoxyl alpha-D-glucopyranoside
• Synonyms: (2R,3R,4S,5S,6R)-2-[(5-bromo-4-chloro-1H-indol-3-yl)oxy]-6-(hydroxymethyl)oxane-3,4,5-triol
• Molecular Formula: C₁₄H₁₅BrClNO₆
• Molecular Weight: 408.63
• CAS Registry Number: 108789-36-2
• SMILES: C1=CC(=C(C2=C1NC=C2O[C@@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)O)Cl)Br

Properties

• Density: 1.9±0.1 g/cm³ Calc.^*
• Boiling point: 673.9±55.0 ºC 760 mmHg (Calc.)^*
• Flash point: 361.3±31.5 ºC (Calc.)^*
• Index of refraction: 1.732 (Calc.)^*

^* Calculated using Advanced Chemistry Development (ACD/Labs) Software.

Safety Data

• Hazard Symbols: GHS07 Warning
• Hazard Statements: H302-H312-H332
• Precautionary Statements: P261-P264-P270-P271-P280-P301+P312-P302+P352-P304+P340-P330-P363-P501

Discovory and Applicatios

5-Bromo-4-chloro-3-indoxyl α-D-glucopyranoside is a synthetic chromogenic substrate designed for the detection of α-glucosidase activity in biochemical and microbiological assays. It belongs to a class of glycosidic compounds in which a halogenated indoxyl moiety is glycosidically linked to a sugar unit, specifically α-D-glucose. This compound is structurally analogous to substrates developed for enzymatic colorimetric reactions, enabling visual detection of enzymatic hydrolysis by the release and oxidation of the indoxyl component.

The development of 5-Bromo-4-chloro-3-indoxyl α-D-glucopyranoside stemmed from ongoing efforts in enzymology and diagnostic biochemistry to provide reliable substrates for distinguishing enzymatic activity of α-glucosidases. These enzymes are involved in the hydrolysis of α-glucosidic linkages in oligosaccharides, polysaccharides, and synthetic substrates. The compound was synthesized as part of a broader group of indoxyl glycosides to serve as colorimetric substrates for specific glycosidase activity detection in complex biological samples.

The mechanism of action involves enzymatic cleavage of the glycosidic bond by α-glucosidase, releasing the 5-bromo-4-chloro-3-indoxyl aglycone. This liberated indoxyl derivative subsequently undergoes spontaneous oxidation in the presence of atmospheric oxygen, forming an intensely colored blue indigo dimer. The visible change in color provides a direct, sensitive method for determining the presence and activity of α-glucosidase. This property has made the substrate a preferred choice in microbial screening, tissue histochemistry, and biochemical enzyme assays.

5-Bromo-4-chloro-3-indoxyl α-D-glucopyranoside has been widely adopted in microbiology for the selective identification of microorganisms that express α-glucosidase. In differential media, such as chromogenic agar plates, the substrate is incorporated into the growth medium, and colonies expressing the enzyme can be visually distinguished by the formation of blue precipitates. This technique is particularly valuable in clinical microbiology for differentiating bacterial species based on their enzymatic profiles.

In enzymology, the substrate is employed in solution-based assays where α-glucosidase activity needs to be quantified. Because the reaction product is insoluble and highly colored, it can be measured spectrophotometrically or detected visually. This allows for flexible applications ranging from qualitative assessments in screening assays to quantitative enzyme kinetics studies. The substrate has also been applied in monitoring enzymatic degradation in lysosomal storage disorder research and in studying carbohydrate metabolism pathways.

The compound is suitable for use under mild physiological conditions, typically at a neutral to slightly acidic pH, which aligns with the optimal activity range of many α-glucosidases. It is compatible with a variety of biological matrices, including cultured cells, tissue sections, and microbial suspensions. Assay protocols using this substrate often include incubation at controlled temperature and pH, followed by observation of the colorimetric change resulting from the formation of the oxidized indigo product.

Chemically, 5-Bromo-4-chloro-3-indoxyl α-D-glucopyranoside is stable when stored under dry and cool conditions. It is generally supplied as a powder and is soluble in organic solvents or buffered aqueous solutions, depending on the application. Its halogenated indoxyl structure enhances oxidative dye formation and contributes to the high contrast of the visual readout. The compound's specificity for α-glucosidase over β-glucosidase makes it a useful tool for selective enzymatic analysis.

The practical significance of this compound lies in its reliability, high sensitivity, and the clarity of the visual output in detecting enzymatic reactions. It is used both in research laboratories for enzyme characterization and in clinical settings for microbial diagnostics. Its role in advancing chromogenic substrate technology continues to support the broader applications of enzymatic detection in biological and analytical sciences.