Electrophoresis separates charged molecules in an electric field based on size, charge, and shape.
Traditionally used for proteins and nucleic acids, it is increasingly applied to small-molecule analysis in crude drug characterization.
1) Polyacrylamide Gel Electrophoresis (PAGE)
Principle
Molecules migrate through a polyacrylamide gel under an electric field.
Application
Separates complex protein or peptide mixtures in biological samples.
Detects protein-based contaminants or active components in botanical extracts.
2) Capillary Electrophoresis (CE)
Principle
Uses narrow capillaries where molecules move under an electric field with high efficiency.
Application
High-resolution separation of charged small molecules, peptides, and drug components.
Rapid, low-volume analysis for quality control and high-throughput screening.
Coupled with UV or MS detectors (CE-MS) for enhanced identification.
3) Isoelectric Focusing (IEF) (Application of electrophoresis)
Principle
Separates molecules by their isoelectric point (pI) in a pH gradient.
Application
Characterization of proteins and enzymes in crude extracts.
Purification of specific enzymes or protein-based therapeutics from natural sources.
Electrophoretic techniques complement chromatography and spectroscopy, enhancing the characterization of bioactive compounds.