High Performance Liquid Chromatography: A Technical Perspective

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We’ve already discussed how scientists use high-performance liquid chromatography (HPLC), and why it’s important during the process of making discoveries. Suppliers also have a significant role in this process, since they provide scientists with the tools they need. At Strategic Directions, the analytical market forecasting arm of BioInformatics Inc., our intention is to inform businesses about the entire market for HPLC, including every associated instrument, consumable, or service. Once you’ve acquired this report, you’ll have access to an exhaustive conglomeration of data, analyses, and commentary, all of which pertain to the market dynamics of various HPLC instrumentation. We know that this data is especially comprehensive, since we’ve analyzed markets in chromatography that aren’t detailed in our Global Assessment Report. Let’s take a quick look at the multitude of techniques that are featured in  HPLC 2018: Market Analysis and Perspectives.

Ultra-High-Performance Liquid Chromatography (UHPLC) is a sample preparation technique that utilizes smaller particle sizes, which leads to much higher resolution and increased sensitivity. Additionally, the time it takes to analyze sample solutions is drastically reduced. Smaller particle sizes lead to increased backpressure, but this is counteracted by pumping the mobile phase at higher pressures or lowering the temperature of the liquid phase. Most UHPLC systems can withstand up to 1,300 bar of pressure.

Micro/capillary/nano liquid chromatography systems are often used for biomolecular analysis. These instruments are differentiated by their column internal diameter and flow rate. For example, nano liquid chromatography typically involves a column with an internal diameter of 10-100 mm and utilizes flow rates of 10-1000 nl/min. Often described as miniaturized HPLC, these systems have lower flow rates to maximize analytical sensitivity. Metabolomics and proteomics are major areas of study where these instruments are commonly used.

Size Exclusion Chromatography (SEC) is a technique used to separate sample components based on differences in size, so it is best suited for the separation of oligomeric and polymeric samples. The column of SEC instruments is slightly different from other chromatography systems, because it contains porous compounds, such as cross-linked gel. As the sample flows through the column, smaller molecules become trapped in the column, while larger molecules flow through the column unimpeded. A detector quantifies the weight and concentration of these molecules.

Amino Acid Analyzers (AAA), as the name implies, are designed to separate and analyze amino acids. Prior to chromatography, hydrolysis is typically used to break the peptide bonds between adjacent amino acids. All amino acids have the same general structure: an amino group, a center carbon with an attached side chain, and a carboxylic acid group. The side chain, and its specific chemical structure, gives amino acids different properties in different biological environments. AAA attempts to make use of these differences by employing columns with different polarity, hydrophobicity, or charge.

Preparative High-Performance Liquid Chromatography (prep HPLC) systems are mainly used to isolate components of a sample instead of providing measurements. Generally, this equipment operates at higher flow rates, uses large columns, and employs larger particle sizes. Although its fairly expensive, prep HPLC is very useful because of its specificity and usability. Some of its key applications include the isolation of  biologics and pharmaceutical compounds.

Super Critical Fluid Chromatography (SFC) is a distinct technique because the mobile phase is neither liquid nor gas. Super critical fluids have been compressed and heated past its critical point, so it displays the low viscosity of gas and the high density of liquid. The high density of super critical fluids maximizes solvation strength. Extremely similar molecules, like enantiomers or chiral compounds, are more easily separated using super critical fluids. The low viscosity of super critical fluids provides high diffusivity rates and allows the use of longer columns. Additionally, the mobile phase can be recycled, which increases efficiency and reduces costs. Carbon dioxide is most commonly used as the solvent because of its low critical temperature and compatibility with mass spectrometry.

Chromatography instruments collect much data quantifying various sample components. In order to correctly process, filter, and store this data many scientists use chromatography data systems (CDS). These software programs enable users to control multiple instruments simultaneously and allows multiple users to access data in a secure network. Currently, workstation and client/server CDS programs are the most common types. The latter offers multi-user capability, and often maintains a dedicated server networked to several PCs at once.

In addition to analyzing the current status of various markets in chromatography, we’ve included much data and predictions concerning the future of chromatography. 5-year forecasts will help suppliers make important changes such as increasing the inventory of specific consumables, or focusing their marketing strategy on scientists in a particular sector. Businesses can also utilize these predictions to perform internal assessments and identify points of future improvement.


By purchasing this report, businesses will equip themselves with the necessary knowledge to become a major player in the chromatography market. Stay on the lookout for the final post in this series, where we will provide sneak previews into the content of the upcoming full-length report: HPLC 2018 Market Analysis and Perspectives.

2 replies
  1. Maria Brown
    Maria Brown says:

    Wonderful article about technical aspect of High Performance Liquid Chromatography. Keep sharing these types of articles.

  2. Jeff Carbine
    Jeff Carbine says:

    I loved when you said that chromatography instruments collect much data quantifying various sample components. In order to correctly process, filter, and store this data many scientists use chromatography data systems (CDS). Liquid chromatography is instrumental, especially in this time of the pandemic. We need these tools not only in the labs but in our homes as well. I will consider purchasing one soon.

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