High-performance liquid chromatography (HPLC)

HPLC Analysis

HPLC analysis, or High Efficiency Liquid Chromatography, is suitable for the analysis and separation of chemical and botanical compounds and is used in various fields of chemistry, pharmacy, industrial chemistry, and clinical diagnostics. HPLC analysis, or High Performance Liquid Chromatography, has been the fastest growing of all analytical separation techniques with annual sales in the billions of dollars range. Qualitative analysis is performed to detect and identify the type of compounds.

HPLC analysis, or High Performance Liquid Chromatography (sometimes called High Pressure Liquid Chromatography), HPLC analysis is a chromatographic method that can separate mixtures of compounds and is used in biochemistry and analytical chemistry to quantify and purify components of a mixture. HPLC analysis typically uses various types of stationary phases, a pump that moves the mobile phase(s) and analyte through a column, and a detector to provide a possible retention time for the analyte. The detector may also provide additional information related to the analyte, (i.e. UV/Vis spectroscopic data for the analyte if equipped). The retention time of the analyte varies depending on the strength of its interaction with the stationary phase, the ratio/composition of the solvent(s) used, and the flow rate of the mobile phase. This is a type of liquid chromatography that uses a smaller column size, smaller volume of the solvent inside the column, and a higher pressure of the mobile phase.

The pump provides higher pressure to move the mobile phase and analyte through the dense column. The increased density results from the smaller particle size. This allows for better separation in columns of shorter length compared to conventional column chromatography. The sample to be analyzed or separated is introduced into the mobile phase flow in a small volume. The solute transported through the column is reduced by specific chemical or physical interactions with the stationary phase present in the column. The velocity of the solution depends on the nature of the sample and the composition of the stationary phase (column).

The time it takes for the sample to exit the column is called the retention time. Retention time is considered a characteristic of a given sample under certain conditions. Using a column with a smaller particle size (which results in lower pressure) increases the linear velocity, allowing less time for diffusion through the column and improving the clarity of the chromatogram. Common solvents used include a soluble mixture of water or various organic liquids (the most common are methanol and acetonitrile). The water may contain a buffer or salt to separate the sample components, or compounds such as trifluoroacetic acid that act as an ion-pairing agent.

Applications of HPLC Analysis

• Food and Flavoring Analysis
• Rapid Screening and Component Analysis of Soft Drinks
• Multinuclear Analysis of 301 Pesticides in Food Samples by HPLC-MS/MS
• Environmental Analysis
• For Pharmaceuticals and Personal Care Products in Water, Soil, Sediment and Biota by HPLC/MS/MS
• Rapid Separation and Identification of Carbonyl Compounds by HPLC
• Specialized Chemical Analysis
• Analysis of Phenolic Antioxidant Compounds and Urocamides in Polymers Using Fast LC
• A Complete Solution for the Analysis of Melamine and Cyanuric Acid in Pet Food by GC/MS and Natural Phase Aqueous LC/MS/MS
• Forensic Analysis
• Determination of Benzodiazepines in Edible Fluids Using LC/MS/MS
• Pharmaceutical Impurity Profile Analysis
• Impurity Profile with LC System
• Discovery Analysis of Pharmaceutical Compounds