UNDERSTANDING UHPLC TRAINING COURSE
This course explores the key principles and practical considerations of Ultra High Performance Liquid Chromatography (UHPLC) to facilitate robust methods suitable for routine analysis. Covering essential topics such as system hardware requirements and differences, column design and chemistry, flow paths, and performance-critical parameters like resolution, efficiency, selectivity, and backpressure.
A strong emphasis is placed on practical considerations, unique to UHPLC, that cause issues with method robustness in routine applications. These include considerations during method development, such as mobile phase composition, column chemistry and dimensions, to day-to-day considerations such as differences in hardware and preparation of reagents and samples.
With a clear focus on practical applications, this training enhances understanding and confidence in UHPLC, helping participants troubleshoot effectively, optimize methods, and make better-informed decisions across the entire chromatographic workflow. By building both theoretical insight and practical skills, the course promotes greater independence, reduced errors, and improved return on analytical investment.
UNDERSTANDING UHPLC COURSE DETAILS
- Chromatography fundamentals and system components
- Key performance metrics -resolution, efficiency, and selectivity - what they mean and how to optimize them.
- Mobile phase optimization strategies including flow rate, additives, gradients, pH control, and preparation best practices.
- Column hardware. Impact of tubing, dimensions, and fittings on performance and pressure.
- Column chemistry. Packing types, particle size, temperature effects, and efficiency factors.
- UHPLC system hardware. Pump and autosampler designs, detector options, and flow cell considerations.
- System performance factors. How hardware choices influence reproducibility, dwell volume, and transferability.
- Practical insights into optimizing and maintaining your setup
We offer flexible delivery options for this instructor-led course:
- On-site: one day at your lab or a location of your choice.
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Online: two half days.
View our online training calendar for upcoming dates.
Previous experience is not essential however delegates would benefit from a basic understanding of HPLC and some experience in routine HPLC analysis.
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Introduction
- Chromatography foundation
- Polarity and basic molecule properties
- Modular HPLC system
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Describing Chromatography
- Resolution
- Capacity factor
- Efficiency
- Selectivity
- UHPLC practical considerations
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Mobile Phases
- Flowrate
- Organic additives
- Gradient elution
- Ionisation, pH control and buffering
- Mobile phase preparation for UHPLC
- Ion pairing reagents
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HPLC Column Hardware
- Tubing and fittings
- Column length
- Internal diameter
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HPLC Column Packing
- Silica packing
- Porosity
- Particle size
- Advantages and disadvantages of sub 2 µm column packing
- Factors other than column affecting efficiency
- Back pressure
- Superficially porous particles
- Column temperature
- Bonded silica – endcapping, self-association, pH restrictions
- Chemically bonded ligands
- Column protection and repair
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HPLC System Hardware
- Pump designs – pump designs, components, high pressure mixing and low pressure mixing
- Autosampler designs – designs, switching valve, components
- UV / PDA detection, flow cell requirements for UHPLC
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Hardware Differences
- Pump
- Gradient dwell
- Extra column volume
- Detector speed
- Gradient elution – worked example
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Is UHPLC Ideal For Your Application?
- Particle size
- Sample preparation costs
- Solid core
- Reduced solvent usage
