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Certificate in High Performance Liquid Chromatography
Training by Distance Learning
 Section 1: Theory
This section covers the fundamental principles behind the science of chromatography. It provides an understanding of what is happening in a separation along with how to assign meaningful values to some of the more important parameters, values which can be of great benefit when assessing the performance of HPLC systems. This section has ten associated assignments and covers the following topics:
- What is chromatography?
- The different modes of liquid chromatography
- Peak broadening
- Efficiency
- Retention factor
- Plate theory
- The Van Deemter equation, A-term, B-term, C-term
- H-U curve
- Resolution
- Selectivity factor
- Peak shape, measurement and implications
 Section 2: Instrumentation
Understanding the component parts of a HPLC system is vital in getting the best from any instrument, and knowing the contribution each makes to the overall analysis is invaluable in troubleshooting and method development. This section has ten associated assignments and covers the following topics:
- The basic HPLC system
- The solvent delivery system, including different types of pumps
- Sample introduction, including types of injector
- Problems associated with autosamplers
- The column oven/chiller and the influence of temperature
- The influence of temperature on efficiency
- The influence of temperature on selectivity
- The effect of temperature on back pressure and stationary phase
- Detectors and important associated specification parameters
- Types of detector including UV/Vis, RI, fluorescence and MS
- Connecting tubing and associated fittings
 Section 3: The column
How does the chromatographer know which column to choose and how to use it? This section helps to make sense of all the different types, which to use when, how to use them and how to look after them. This section has ten associated assignments and covers the following topics:
- Silica and normal phases, liquid phases, bonded phases
- End capping
- pH considerations
- Polymer based packing
- The silica surface, surface blocking and embedded groups
- Specialist packings: carbon, HILIC and chiral
- Particle size, pore size and column dimensions
- Guard columns and column washing
- Introduction to narrow-bore columns
- Solute dispersion in the column
- Sensitivity and solvent usage with narrow-bore columns
- Reducing extra-column dispersion
- Injection in narrow-bore columns
- Gradient elution with narrow-bore columns
 Section 4: The mobile phase
One of the great strengths of HPLC is the choice of mobile phases available and the changes in selectivity these can provide. But which should be used and how should they be prepared? The dos and don'ts are explained in this section. This section has twevle associated assignments and covers the following topics:
- Solvent purity and why it is so important
- Mobile phase degassing and filtration
- Determining the mobile phase for a separation
- Mobile phase pH
- Gradient elution
- Column equilibration between gradients
- Scaling gradients, extra column effects and dwell volume
- Mobile phase viscosity
- The mechanism of ion-pair chromatography
- Developing methods with ion-pair reagents
 Section 5: Integration & Quantification
Understanding integration and the limits of integrating systems is vital for generating accurate results. Quantification often forms the final part of the analysis, generating real results from the chromatography achieved. This section guides the candidate through integration and types of quantification routinely used, explaining advantages and disadvantages of each. This section has eight associated assignments and covers the following topics:
- Introduction to integration and detector contribution
- Poorly resolved peaks
- Errors created by the baseline
- Tangent skim integration
- Introduction to quantification
- % peak area quantification
- % peak area normalised quantification
- External standard quantification
- Internal standard quantification
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