HPLC is applied to separation and quantitation of components of mixture of compounds that may not be stable at higher temperatures encountered in Gas chromatographic separations. The technique has gained wide acceptance in the field of pharmaceuticals, foods & beverages, agricultural products, cosmetics, polymers and synthetic materials and environmental studies.

The programme will be beneficial to

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Introduction to HPLC Course and its Objectives

We thank you for visiting our training site lab-training.com and registering for the free e-learning course on HPLC. We understand that everyone has busy work schedules and today’s hectic life style leaves you little or no time to refer voluminous books to learn any technique. However, for sustained growth learning has to be adopted as a lifelong habit……

Module 1 : General Introduction to Chromatography

Chromatography is the most widely used laboratory technique for separation, identification and quantification of components of liquid and gaseous mixtures. Solid mixtures are also analyzed by first converting them to a liquid or gaseous state, using suitable sample preparation techniques. Differential affinity of components between the carrier and stationary phases forms the basis of separation. Components retained by stationary phase have….

Module 2 : Liquid Chromatography Evolution

Liquid Chromatography originated in early 1900 when Russian botanist Mikhail S. Tswett separated plant pigments using calcium carbonate packed glass columns. It was not until mid century that the technique was applied to develop Paper Chromatography, HPLC and GC.

HPLC was originally referred to as High Pressure Liquid Chromatography as high pressure was required to allow liquid to flow through packed columns. However, with continued advances in instrumentation and packing materials ….

Module 3 : Introduction to High Performance Liquid Chromatography and its parts

Chromatography equipment look rather intimidating to anyone who has not handled them before, but on a closer look and as you get familiar with the equipment you realize that behind the network of wires, complex plumbing and circuitry is a simple machine with only a few major parts. Different combinations of these parts namely pumps, detectors and injectors yield a infinite number of configurations based on the application…..

Module 4 : Types of Stationary Phases

 In Module 1 an analogy was given comparing molecular mixture to a family passing by a candy store which is like the stationary phase. Children get retained because of their affinity for candy while the parents keep on moving like un- retained molecules leading to separation between them. Partitioning of sample molecules between a mobile phase and the stationary phase in the HPLC column is based on affinities which tend to hold back some molecules while allowing others to pass through freely….

Module 5 : Types of Mobile Phases

In the previous module you were introduced to the stationary phases which serve to retain selectively the sample components. After separation these components need to be transported to the detector for detection and quantification. Mobile phase is the life line of HPLC system as it transports the sample from the injector to the detector and its characteristics such as composition, pH etc, have a profound effect on separation of sample components. The mobile phase should have the following desirable characteristics to carry out this important function…..

Module 6: Types of Detectors

A detector can be compared to a gate watchman who verifies the visitors before permitting them entry inside a building. The chromatographic detector is capable of establishing both the identity and concentration of eluting components in the mobile phase stream. A broad range of detectors is available to meet different sample requirements. Specific detectors respond to a particular compound only and the response is independent of mobile phase composition. On the other hand the response of bulk property detectors is dependent on collective………….

Module 7 :Types of Pumps

In the last module you appreciated the crucial role played by a detector in the HPLC system. In this module you will be introduced to the pump which provides continuous and consistent flow of mobile phase through the HPLC system. A pump can be compared to the human heart which continuously pumps blood throughout the body but though the human heart can withstand changes in blood pressure within specified limit due to stress and strain the HPLC pump is required to deliver flow of mobile phase at constant pressure and flow rate……

 Module 8 : Types of High Performance Liquid Chromatography Injectors

Understanding of injectors is as important as that of pumps in the last session. Injectors serve to introduce required sample volume accurately into the HPLC system.

Sample injection into the moving mobile phase stream in HPLC is quite different from injection into a gas stream in Gas Chromatography as precise injection is required against high back pressure. In such a situation it is not possible to simply inject using a syringe alone…..  

Module 9 : Applications of High Performance Liquid Chromatography 

After having gained exposure to High Performance Liquid Chromatography systems and their components we now introduce to typical applications.

HPLC has contributed to analytical solutions in diverse fields such as pharmaceuticals, foods, life sciences, environment, forensics, etc. In the present module we shall discuss some application areas in pharmaceuticals and foods…….

Module 10  : Top 10 Interview questions on High Performance Liquid Chromatography

Your understanding and pickup of a topic is gauged by your ability to answer questions related to basics and practical aspects. We now provide you an opportunity to familiarize yourself with some typical questions that you may face in job interviews involving extensive use of High Performance Liquid Chromatography systems….