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Frequently Asked Questions Regarding High-Performance Liquid Chromatography

High-performance liquid chromatography (HPLC) has been used in different industries that mostly deal with chemicals. For instance, HPLC has been used in the manufacturing industry during the process of making biological and pharmaceutical products. It has also been used for many medical purposes during lab tests such as detecting vitamin D levels in blood serum.

HPLC is also a technique that is used in sports for the detection of performance enhancement drugs in urine and in research for separating components in complex biological samples. Despite its significance, there are still individuals who are unaware of HPLC. This article will discuss  what is HPLC, and answer the frequently asked questions about it.

What is HPLC?

HPLC was formerly known as high-pressure liquid chromatography. According to an article by Chem Guide, HPLC is an improved form of column chromatography, which is a technique used to identify, separate, and quantify components in a given mixture. HPLC methods are highly-automated and extremely sensitive.

So what is HPLC in simple terms? HPLC is a better technique because instead of allowing a solvent to drip through a column, HPLC forces the solvent under high pressures, which makes the process faster. HPLC allows the utilization of much smaller particle sizes, which provides a greater surface area for interaction during different phases.

What Are the Different Types of HPLC?

The majority of chromatographic separations operate under the same principle, which is the separation of particular components on a given sample. However, there are variations of HPLC, which include the following.

Normal HPLC. This technique separates components based on their polarity. This is done by using a polar stationary phase that uses silica and a non-polar mobile phase that uses methylene chloride, hexane, or a mixture of these components.

Size-exclusion HPLC. This HPLC variation uses material that has precisely-controlled pore sizes, which separate components according to their molecular and particle sizes. Small particles penetrate inside the porous material, while large molecules are washed through the column.

Ion-Exchange HPLC. This technique is mostly used with ionic compounds, hence the name. The stationary phase of this method uses an ionically-charged surface. The stronger the charge, the better it will attract to the ionic surface. On the other hand, the mobile phase uses an aqueous buffer that uses ionic strength and pH to control elution.

Reverse HPLC. This technique has the same principles as a normal HPLC. However, it uses different phases. The mobile phase uses a polar liquid, while the stationary phase uses a non-polar liquid or hydrophobic substance.

What are the Benefits of HPLC Systems?

HPLC systems have several benefits and advantages, which can undoubtedly help your company or institution to have a more efficient and effective laboratory. Some of these benefits include the following:

Test Both Raw Materials and Finished Products

HPLC technique can be done for several types of raw materials, which can help you decide what material would be the best to use for a particular product; that you want to develop. You can also test finished products for contaminants or other impurities to ensure that you release a safe product in the market.

Gain Competitive Advantage

HPLC can be used to perform analyses of the products of your competitors. This means you will be gaining a better understanding of the different products of different companies in your industry.

By doing this, you will be able to tweak your product to make it better among the rest of your competitors, which will make you stand out. By using HPLC, you will gain the upper hand in your industry and have a competitive advantage.

A Reliable and Affordable Technique

The greatest advantage of HPLC, among other techniques, is that it is reliable. Because of this, HPLC can help you solve product failure problems so that you can develop better products for your company. Also, HPLC testing can be done even with just a small sample size, making it an affordable method to test numerous products.

Improves Productivity

HPLC systems improve productivity as it helps in the majority of the areas of analysis, from sampling to instrumentation, separation, and reporting. HPLC systems will provide your laboratory with data management, security features, and validation.


HPLC as a technique is, without a doubt, beneficial and advantageous. This is why HPLC systems should be a part of your institutions’ laboratory. Also, HPLC as a method is something that you should learn because having this knowledge in your arsenal will make your work more effective and productive.

Author: Ester Adams