HPLC
High-Performance Liquid Chromatography — the gold-standard analytical method for measuring peptide purity.
Definition
High-Performance Liquid Chromatography, an analytical technique used to separate, identify, and quantify components in a mixture. In peptide testing, HPLC is the gold standard for measuring purity by separating the target peptide from impurities.
In More Detail
HPLC pushes a sample through a column packed with stationary-phase particles using a high-pressure pump. As the sample travels through the column, each component interacts differently with the stationary phase and elutes at a different time. A UV detector measures absorbance and produces a chromatogram, where the area of each peak corresponds to the relative amount of each compound. For peptides, reversed-phase HPLC (RP-HPLC) with C18 columns and water/acetonitrile gradients is the standard. Purity is reported as the percentage area of the main peak relative to the total integrated area at 214 nm or 220 nm.