Going Beyond Theory And Books: High-performance Liquid Chromatography

The basic principle of scientific study is research and analysis. Science helps further understand the world so that all beings may attain their potential while co-existing harmoniously. The methods used in scientific research have developed technically and technologically over the years, like HPLC, GC, LCMS, etc., which are different types of Chromatography, a popular scientific research technique.

Chromatography is a technique that studies organic compounds. This technique essentially separates an organic mixture into its separate parts using two constituents – a stationary phase and a mobile phase. A lab technician first dissolves the organic mixture in the solvent or the mobile phase. Once the organic mixture dissolves in the mobile phase, they pour or release it over the stationary phase. Each component of the mixture has unique characteristics and thus reacts with the surface of the stationary phase differently. As the mobile phase moves over the stationary phase, the mixture’s components separate from the mobile phase. Since each component reacts differently, they separate at different locations over the stationary phase. Each separation occurs as a swatch of colour.

Scientists and researchers study these swatches to understand the particular component and the organic mixture itself. This method of analysis is the basic principle of Chromatography, founded in 1900. Today, improvements to Chromatography exist. One such advanced technique is HPLC or High-Performance Liquid Chromatography.

THE LABORATORY TECHNIQUE FOR THE 21st CENTURY

People working in chemistry labs will have an in-depth knowledge of Liquid Chromatography because it is one of the most often-used laboratory techniques. HPLC is a type of Chromatography that uses more instruments and is completely liquid. Earlier Chromatography relied on paper to separate the different components of the organic mixture, but Liquid Chromatography uses an adsorbent stationary phase instead.

Essentially, Liquid Chromatography can separate organic mixtures that are nonvolatile, with heavy molecular structures, and are unstable thermally. Liquid Chromatography uses liquid solvents as the mobile phase, into which the technician dissolves the organic mixture. They then put this liquid under high pressure and pass it through a column that contains the adsorbent material or the stationary phase. This method was earlier referred to as “High-Pressure Liquid Chromatography” and later changed to “High-Performance”.

When the high-pressure liquid solvent passes through the adsorbent material, the different compounds react differently with the adsorbent material and separate onto it. Like the basic principles of Chromatography, the components have unique characteristics and respond differently to the stationary phase and alter the flow of the high-pressure liquid. The components separate on different levels of the adsorbent material, depending on their characteristics. The technicians then individually study these components.

Today, this method has a wide range of uses and dedicated software and system. After much scientific research, it has become possible to computerise the entire Chromatography process. This process now has minimal manual requirements, allowing for faster and accurate results. There are even specific courses that train technicians to use this method and the software efficiently. This method reflects the technological advancements in science.

USING HPLC: APPLICATION IN DIFFERENT FIELDS

High-Performance Liquid Chromatography is a versatile technique with vast opportunities for application in different fields. The complete computerisation of this technique makes it easier for people to use, even if they don’t have a scientific background. Today, this technique is regularly put to use in the following ways,

In Research

As with every laboratory technique, technicians primarily use Liquid Chromatography for scientific research. This method can help identify the different chemical components of naturally occurring mixtures or compounds. By separating the various components, scientists can study each part separately and gain a deeper understanding of the compound, its origin, its characteristics, uses, etc.

In Manufacturing

Pharmaceutical companies are the primary users of Liquid Chromatography for manufacturing. They used this method to test the purity of pharmaceutical drugs. This method is highly accurate and can effortlessly detect any abnormalities or impurities in the chemical structure of the pharmaceutical drug. This technique ensures that the company only produces the highest quality of drugs.

For Legal Tests

Liquid Chromatography is the primary technique to detect drugs and other narcotic substances in the urine. Technicians administer these tests to athletes before major tournaments and even for other legal cases that require drug tests. This Chromatography separates the drug components from the urine components on the adsorbent material displaying accurate results.

In Medical

HPLC is nearly unparalleled in its accuracy of findings, which is why it has extensive application in the medical field. One central use of this method is analysing blood samples. Like urine, Liquid Chromatography can separate the different blood components and help identify any abnormalities, impurities, or even deficiencies. Even today, this technique helps lab technicians find Vitamin D deficiencies in the blood.