What is the Origin of Reverse-Phase HPLC?

The term combines 'Reverse-Phase', referring to the stationary phase being non-polar and the mobile phase being polar, with 'HPLC', an acronym for High-Performance Liquid Chromatography. This separation principle is the inverse of traditional normal-phase chromatography. RP-HPLC is a standard and necessary tool in quality control and advanced clinical research laboratories worldwide.

What is the Mechanism of Reverse-Phase HPLC?

The separation mechanism relies on the differential partitioning of analytes between the non-polar stationary phase, typically C18-bonded silica, and the polar mobile phase, which is a mixture of water and an organic solvent. Hydrophobic compounds spend more time interacting with the non-polar stationary phase and thus elute later, while more polar compounds are carried faster by the mobile phase. By precisely controlling the mobile phase composition, RP-HPLC achieves the necessary separation of chemically similar hormonal species for accurate quantification.

Reverse-Phase HPLC

Meaning

Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) is a highly sophisticated analytical chemistry technique employed for separating, identifying, and precisely quantifying components in complex chemical mixtures. In the realm of hormonal health, it is an essential tool for accurately measuring various steroid hormones and their metabolites, as well as ensuring the purity and potency of pharmaceutical preparations. This method offers high resolution and sensitivity for separating compounds based primarily on their differential hydrophobicity.