Quality control, research and production in chemistry require efficient ways to separate components in a compound. Preparative HPLC (High Performance Liquid Chromatography) focuses on separating components in compounds at the molecular level. The main goal is to isolate and purify a significant amount of a target compound from a sample.
Laboratories worldwide use Preparative HPLC. In combination with other technologies, Preparative Chromatography results in highly efficient and reliable compound separation. Unlike Analytical HPLC, where the main goal is quantifying and qualifying components in a mixture, Preparative HPLC aims to purify enough quantities of a particular substance for further use.
From a research perspective, isolating active compounds aids in the study of their structure and properties. Preparative HPLC helps in quality control, ensuring products meet set standards of purification of target compounds. Prep HPLC also can be a step in production, isolating compounds used as ingredients in products.
Role of Components and steps in Preparative HPLC
The analytical column or Prep HPLC column’s primary function is to contain a substance for the separation of mixture components using differences in their physical or chemical properties. The Prep HPLC column is packed with a material, often silica gel, that is known as the "stationary phase." Factors like the particle size and characteristics of the Preparative HPLC stationary phase define the separation efficiency.
The compound to be purified is pushed through the column under pressure using the applicable solvent (the "mobile phase"). The separation of the compounds in the mixture is based on the affinity of each component’s molecules for the mobile and stationary phases. The varied mass of the molecules and their varying propensity to adsorb, or adhere to, the stationary phase causes the compounds to migrate at different speeds, creating separate bands of compound ingredients.
The Preparative HPLC process begins with introducing a sample into the Prep HPLC column under high pressure. As the sample travels through the HPLC column, the different components separate based on their interaction with the stationary phase. Each compound's elution time -- the time it takes to pass through the column -- is recorded. The pure target compound is then collected for further use or analysis.