High-performance liquid chromatography (HPLC) is a robust bioanalytical tool to separate ions, organic molecules, and proteins. Adsorption, ion exchange, and partition are the primary mechanisms in HPLC analysis. HPLC usually involves a solid stationary phase and a liquid mobile phase. The differences in relative distribution solute ratio between the stationary and mobile phases separate the molecules of interest. This characteristic has made HPLC lab employ this technique to assess the purity and content analysis of pharmaceutical molecules. The current article focuses on understanding HPLC labs and their impact on the scientific world.
HPLC Analysis Revolutionizing Scientific Studies
Today researchers have several bioanalytical techniques for assessing drug compounds, such as MSD assays. MSD is a robust technology and offers several assay formats, such as Mesoscale cytokine multiplex assay. HPLC analysis is one such technique.
The primary principle behind HPLC analysis is that some components within a mixture take longer to elute from the chromatography column. The time taken to pass from the chromatography column depends on the molecule’s affinity with the mobile and stationary phases. Molecules with higher affinity will take an extended period to elute from the columns.
HPLC systems are essential for separating, identifying, purifying, and quantifying several compounds. Hence HPLC analysis has numerous applications. Let us explore some of the applications of HPLC analysis.
In clinical sciences and drug development, HPLC systems are used widely to quantitate antibiotics. For example, researchers employed HPLC to quantitate desactylcephalothin and cephalothin in serum samples. Researchers employed reversed-phase chromatography to separate these drugs and then detected them using ultraviolet light absorption. The serum concentration of these antibiotics was as low as one jig/ml. They concluded that HPLC is a specific, sensitive, rapid, and accurate technique for routine clinical investigations.
Similarly, in a different study, researchers employed HPLC to analyze detergent, solubilized intact mass and purified integral membrane proteins. Even here, they used reversed-phase columns to separate detergents and member membrane proteins chromatographically. HPLC analysis provided a 10X lower sensitivity. Besides, this assay is compatible with the hyphenated liquid chromatography-mass spectrometry technique. The researchers concluded that HPLC analysis can be approached via a multi-user instrument or applied in high throughput workflows for evaluating and analyzing membrane proteins. Let us now focus on the future of HPLC labs.
HPLC systems have been a defining tool for the last four decades, and their importance and applications will likely increase. Microbore and fast columns are the future for analytical applications in biologicals and gene products. As the need for ultra-purification rises, immunoaffinity and affinity techniques will be more frequently needed to produce biotechnological pharmaceuticals. However, higher capacity and more accurate chiral separation columns will be required to optimize the efficiency of pharmaceutical compound mass production.
In the future, researchers may opt for more organic resins. Besides, multiple detectors and computerized optimization techniques will become standard per system. Eventually, robotics will increasingly handle hazardous samples such as viral and bacterial samples, AIDS samples, contaminants, and radioisotopes.
In conclusion, despite the applications developed, HPLC systems will continue to evolve from both preparative and analytical perspectives.