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Determination of Chondroitin Sulfate A with Crystal Violet by Resonance Rayleigh Scattering Technique (CAT#: STEM-ST-0056-YJL)

Introduction

Chondroitin sulfate (CS) is a glycosaminoglycan consisting of repeating uronic acid (D-glucuronic acid or L-iduronic acid), N-acetylgalactosamine (GalNAc) sulfate disaccharide units [-UroA(β-1,3)-Gal NAcS(β1,4)]n. According to the variety of uronic acid or N-acetylgalactosamine sulfate, CS can be divided into several kinds, such as chondroitin sulfate A (CSA), chondroitin sulfate B (CSB), chondroitin sulfate C (CSC), etc, in which CSA and CSC are the major ingredients in almost any animal species and tissue. <br />Crystal violet (CV) is a basic triphenylmethan dye, which exist as a cationic species in near neutral solution. The intensity of RRS can be greatly enhanced by the interaction between CSA and CV and that a new RRS spectrum is produced owing to the formation of CSA-CV supramolecular complex by electrostatic gravitation and hydrophobic force.




Principle

Resonance Rayleigh scattering (RRS) is similar to Rayleigh scattering in nature. Resonance Rayleigh scattering is a special elastic scattering produced when the wavelength of Rayleigh scattering (RS) is located at or close to its molecular absorption band. The key to generating RRS is: when the scattering is at or close to the absorption band of the scattering molecule, since the electron absorbs the electromagnetic wave at the same frequency as the scattering frequency, the electron strongly absorbs the photon energy due to resonance and re-scatters. Its scattering intensity is several orders of magnitude higher than that of pure Rayleigh scattering, and it no longer obeys the Rayleigh law of I∝λ-4. This absorption-rescattering process is called resonance Rayleigh scattering (RRS).

Applications

Resonance Rayleigh scattering is used to the study of aggregation of chromophores on biological macromolecules and the determination of biological macromolecules such as nucleic acid, proteins and heparin, further, it has been used in the determination of trace amounts of inorganic ions and the cationic surfactant by means of ion association reactions with some dyes. In addition, it has been applied to the study of nanoparticles in liquid and the determination of β-cyclodextrin inclusion constant and the critical micelle concentration of surfactant.

Procedure

1. Sample preparation
2. Measurement by scattering detection instrument
3. Data analysis

Materials

Rayleigh scattering measurement system