Hydroxypropyl-beta-cyclodextrin enhanced determination for the Vitamin B12 by fluorescence quenching method

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Vitamin B12 (cobalamin) is essential for numerous metabolic processes.
• Accurate quantification of Vitamin B12 is crucial for clinical diagnostics and pharmaceutical quality control.
• Existing methods for Vitamin B12 determination may have limitations in sensitivity or applicability.

Purpose of the Study:

• To develop a novel and sensitive fluorescence quenching method for the determination of Vitamin B12.
• To investigate the mechanism underlying the fluorescence quenching interaction.
• To evaluate the applicability of the method for Vitamin B12 determination in pharmaceutical formulations.

Main Methods:

• Development of a fluorescence quenching assay utilizing erythrosine sodium (ES) and Hydroxypropyl-beta-cyclodextrin (HP-beta-CD).
• Formation of an inclusion complex (HP-beta-CD-ES) that enhances fluorescence.
• VB12 addition leads to fluorescence quenching, with quenching intensity correlated to VB12 concentration.
• Optimization of reaction conditions and determination of analytical parameters.

Main Results:

• A linear relationship was established between the fluorescence quenching value (DeltaF) and Vitamin B12 concentration (c).
• The linear range for VB12 determination was found to be approximately 0.0 to 2.1 x 10(-5) mol/L.
• A sensitive detection limit of 1.8 x 10(-7) mol/L was achieved under optimal conditions.
• The method demonstrated satisfactory performance for determining VB12 in injection samples.

Conclusions:

• The developed fluorescence quenching method provides a sensitive and reliable approach for Vitamin B12 quantification.
• The method is based on the specific interaction between VB12 and the HP-beta-CD-ES inclusion complex.
• This novel method shows potential for routine analysis of Vitamin B12 in pharmaceutical preparations.