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Reverse Phase HPLC

The team of HPLC experts at GL Technologies has worked hard to earn the trust of biopharmaceutical and related industry organizations thorughout the entire state of California with their HPLC calibration and certification needs. Reverse phase high-performance liquid chromatography (RP-HPLC) is a powerful analytical technique that is widely used in the biopharmaceutical industry. It is particularly useful in the purification and analysis of complex biological molecules, such as proteins, peptides, and antibodies. GL Technologies understands the principles of reverse phase HPLC and its application in biopharmaceutical analysis and can help you with your calibration and certification requirements.

To speak with the experts about your Reverse Phase HPLC Calibration needs, please fill out our online form or give us a call!


Principles of Reverse Phase HPLC

Reverse phase HPLC is a chromatographic technique in which the stationary phase is a hydrophobic material, such as a C18 or C8 bonded silica column. The sample is dissolved in a mobile phase that is typically a mixture of water and organic solvents. The hydrophobic stationary phase interacts with the hydrophobic regions of the sample molecules, causing them to be retained on the column. The retention time of the molecules is determined by their hydrophobicity, size, shape, and charge.

In reverse phase HPLC, the escape order of the sample molecules is based on their hydrophobicity, with the more hydrophobic molecules being retained on the column for a longer period of time. The mobile phase composition can be adjusted to improve the separation of the sample components. For example, increasing the organic content of the mobile phase will generally decrease the retention time of the sample molecules.

Reverse Phase HPLC Uses in Research

Reverse phase HPLC is widely used in the biopharmaceutical industry for the analysis of proteins, peptides, and antibodies. One important application of RP-HPLC is in the purification of pharmaceuticals. In this process, the sample is applied to a reverse phase column and eluted with a mobile phase gradient. The individual components of the sample are separated based on their hydrophobicity and collected for further analysis or use.

RP-HPLC is also used for the analysis of the purity and identity of biopharmaceuticals. By comparing the retention time and spectral properties of the sample with those of reference standards, the identity and purity of the sample can be determined. Additionally, RP-HPLC is often used in the quality control of biopharmaceuticals, to verify that the products meet the required specifications.

Another important application of RP-HPLC in biopharmaceutical analysis is in the determination of protein structure. By combining RP-HPLC with other techniques, such as mass spectrometry and nuclear magnetic resonance spectroscopy, the amino acid sequence and post-translational modifications of proteins can be clarified.

To speak with the experts about your Reverse Phase HPLC Calibration needs, please fill out our online form or give us a call!


San Diego Reverse HPLC Chromatography Biopharmaceutical Certification


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