5-Bromo-4-chloro-3-indoxyl choline phosphate,X-Phos-choline

5-Bromo-4-chloro-3-indoxyl choline phosphate is a phospholipase substrate that has been shown to be efficient in the synthesis of phosphatidylcholine from choline. It has been synthesized by reacting 5-bromoindoxyl with chloramine and phosphoryl chloride, followed by reaction with sodium hydroxide. This compound is an efficient synthetic intermediate for the production of phosphatidylcholine.

5-Bromo-4-chloro-3-indoxyl choline phosphate is a derivative of indoxyl, which is commonly used in the research studies related to enzymatic activity, immunology, and microbiology. This molecule is known to hydrolyze enzymatically, providing visual indications of the enzymatic activity. This paper aims to provide an overview of the chemical properties, synthesis, characterization, analytical methods, biological properties, toxicity, safety, and applications in scientific experiments, along with its potential implications and future directions.

Definition and Background:

5-Bromo-4-chloro-3-indoxyl choline phosphate is the substrate molecule that hydrolyzes by cholinesterase enzymes, generating a colored product. In this reaction, choline is extracted from the molecule, leading to the formation of an indoxyl compound that is oxidized to produce a blue color. This reaction is commonly used to detect and quantify the activity of cholinesterase enzymes. 5-Bromo-4-chloro-3-indoxyl choline phosphate is often used in various fields of research including microbiology, enzymology, neurobiology, and drug discovery.

Physical and Chemical Properties:

The molecular formula of 5-Bromo-4-chloro-3-indoxyl choline phosphate is C16H18BrClN2O6P, and its molecular weight is 476.64 g/mol. It is a light yellow powder that is soluble in water, alcohols, and dimethyl sulfoxide (DMSO). Its melting point is 137-139ºC. This molecule has a high UV absorption profile with a maximum absorbance at 592 nm, making it easy to detect in various assays.

Synthesis and Characterization:

5-Bromo-4-chloro-3-indoxyl choline phosphate is synthesized by condensing 5-Bromo-4-chloro-3-indoxyl with choline phosphate. This reaction is carried out in the presence of a coupling reagent, such as dicyclohexyl carbodiimide (DCC), and a catalyst, such as N,N-dimethylaminopyridine (DMAP). The product is purified using column chromatography or recrystallization.

The characterization of 5-Bromo-4-chloro-3-indoxyl choline phosphate can be done using various techniques including ^1H and ^13C nuclear magnetic resonance (NMR) spectroscopy, infrared (IR) spectroscopy, mass spectrometry (MS), and elemental analysis. Further characterization can be carried out using X-ray crystallography to determine its crystal structure.

Analytical Methods:

Several analytical methods are used to detect and quantify 5-Bromo-4-chloro-3-indoxyl choline phosphate. These include colorimetric assays, high-performance liquid chromatography (HPLC), gas chromatography (GC), and capillary electrophoresis (CE). These techniques can also be used to measure the activity of cholinesterase enzymes.

Biological Properties:

5-Bromo-4-chloro-3-indoxyl choline phosphate is a substrate molecule that is used to measure the activity of cholinesterase enzymes. It is widely used in in vitro experiments to evaluate the effect of different compounds on the activity of cholinesterase enzymes. This reaction is also used to identify and quantify the activity of cholinesterase enzymes in blood samples.

Toxicity and Safety in Scientific Experiments:

5-Bromo-4-chloro-3-indoxyl choline phosphate is a moderately toxic compound, and it should be handled with caution. It can cause irritation to the skin and eyes upon contact. Therefore, proper protective equipment should be used while working with this compound. It is recommended to follow the guidelines provided by the local regulatory authorities for the safe handling and disposal of this compound.

Applications in Scientific Experiments:

5-Bromo-4-chloro-3-indoxyl choline phosphate is widely used in various fields of research including enzymology, immunology, microbiology, and drug discovery. It is used as a substrate molecule to measure the activity of cholinesterase enzymes, which has significance in various research areas. It is also used in drug screening assays to evaluate the effect of compounds on cholinesterase enzymes.

Current State of Research:

Recent studies have focused on the use of 5-Bromo-4-chloro-3-indoxyl choline phosphate in the development of biosensors, which can detect cholinesterase activity in real-time. These biosensors have potential applications in the clinical diagnosis of cholinesterase-related disorders, such as Alzheimer's disease and Parkinson's disease. Moreover, studies have also demonstrated the efficacy of 5-Bromo-4-chloro-3-indoxyl choline phosphate in detecting the activity of cholinesterase in environmental samples, which is useful in detecting pesticide contamination.

Potential Implications in Various Fields of Research and Industry:

5-Bromo-4-chloro-3-indoxyl choline phosphate has potential implications in various fields of research and industry including microbiology, enzymology, neurobiology, and drug discovery. It is used in drug screening assays to identify compounds that can modulate the activity of cholinesterase enzymes, which has therapeutic potential in treating cholinesterase-related disorders.

Limitations and Future Directions:

One of the major limitations of using 5-Bromo-4-chloro-3-indoxyl choline phosphate is its moderate toxicity, which restricts its use in in vivo experiments. Moreover, the synthesis of 5-Bromo-4-chloro-3-indoxyl choline phosphate is a complex process that requires special equipment and expertise. Future research should focus on the development of simple and efficient methods for the synthesis of this compound, which can facilitate its wider use in research.

Future directions for the use of 5-Bromo-4-chloro-3-indoxyl choline phosphate include the development of biosensors that can detect cholinesterase activity in environmental samples, which has significance in monitoring pesticide contamination. Moreover, the potential use of this compound in developing diagnostic tools for cholinesterase-related disorders should also be explored. Additionally, further research is warranted to evaluate the efficacy of 5-Bromo-4-chloro-3-indoxyl choline phosphate in drug discovery, specifically in developing inhibitors of cholinesterase enzymes.