Description
Live-On-The-Web: Systematic LC Method Development; Best Practices for HPLC and UHPLC
Who should take this course?
If you’ve ever wished that HPLC method development were more logical and methodical, this is the course for you. Attendees should have at least one year of HPLC experience in the laboratory and at least some involvement in developing new methods or troubleshooting older ones. This course is for chromatographers who want to develop better methods faster and less expensively.
This course is intended for chromatographers with 2-5 years of hands-on experience. Originally developed by Lloyd Snyder and now taught by chromatography experts John Dolan and Tom Jupille, this course sets out a systematic approach to developing robust HPLC methods quickly and efficiently.
What does it cover?
The course begins with a thorough review of basic chromatography parameters and their relationship as the basis for a rational approach to extract as much useful information as possible from a minimum investment in experimental time and effort. It answers questions such as:
- what is the best packing particle size and column geometry based on user needs?
- how do I transfer legacy HPLC methods to UHPLC?
- how do I make UHPLC methods backward compatible with older HPLC systems?
- can I do the separation isocratically or will I need a gradient?
- how do I decide what type of column to use?
- what’s the best solvent?
- how important is pH?
- how do I optimize a gradient profile?
- when to I need to use ion-pair chromatography?
- should I use HILIC or normal-phase instead of reversed-phase?
and much, much more!
How much does it cost?
The standard “Live on the Web” course fee is $947/person. This includes the course and a download of the course notes. Payment via credit card (MC, VISA, or AMEX only).
Buy additional course registrations now and qualify for a 15% discount. Call us at (925) 297-5374 to arrange multiple-registrant pricing.
What do I need in order to take the course?
- A personal computer or tablet (Windows, Mac,iOS,or Android)
- A high-speed (broadband) internet connection
- A phone connection with headset or speakerphone
Here’s how it works:
- After you register we’ll e-mail your course log-in information and password.
- Download and print a hard copy of the course handout (approximately 200 pages).
- About five minutes before the start of the course, log in to www.gotomeeting.com to download the gotomeeting client and check in for the course. Dial the phone number* for the conference call connection that you were given in your confirmation e-mail.
- The course runs a total of six hours in three 2.5-hour sessions.
*Standard long-distance phone rates will apply (approximately $10.00).
What if I can’t make it?
All sessions are recorded and will be available for review for a month after the end of the course; if you miss a session, you can catch up at your convenience. If you must cancel, please call at least 48 hours prior to the start of the course in order to receive a refund. No refund will be made for cancellation within 48 hours of the course.
About the instructors
The course was developed by Lloyd Snyder, John Dolan, and Tom Jupille. These sessions will be presented by Tom Jupille.
What topics are covered?
The course is presented in three 2.5-hour sessions “live on the web”. This course is interactive, so the exact timing will vary somewhat depending on participants’ interests and questions. A typical schedule looks like this:
Section 1. Getting Started
- Setting analytical goals
- Method Development strategies: OFAT vs. QbD
- Selecting a detector
- Estimating LOD/LOQ
- UHPLC vs. HPLC
Section 2. Review of HPLC & UHPLC Basics
- Basic measurements: k’, alpha, N, Rs, TF
- Overall method development strategy: k’, alpha, N (in that order!)
Section 3. Columns
- Evolution of HPLC/UHPLC
- Parameters affecting N
- Particle size effects
- Impact of particle size on hardware
- Totally porous vs. solid-core particles
- Impact of silica purity
- Bonded-phase column chemistry
Section 4. Strategy: reversed-phase of neutral molecules
- Mechanism of reversed phase and its impact on selectivity
- Initial column and mobile phase selection
- Controlling k’
- Controlling alpha: mobile phase strength
- Controlling alpha: temperature
- Controlling alpha: mobile phase type
- Controlling alpha: column type
- Developing orthogonal methods
Section 5. Strategy: reversed-phase of ionizable molecules (acids or bases)
- Effect of pH on retention & selectivity
- Choice of buffer: identity & concentration
- Dealing with tailing problems
- pH – temperature interaction
Section 6. Strategy: Ion-pair & mixed-mode chromatography
- Mechanism of ion pair
- Choice of ion-pair reagent type and concentration
- Problems with ion-pair
- Alternatives to ion-pair: mixed mode columns
Section 7: Strategy: HILIC
- What is HILIC and what is it good for
- Initial column and mobile phase selection
- Controlling k’ (mobile phase water content)
- Controlling alpha: mobile phase strength
- Controlling alpha: mobile phase type
- Controlling alpha: pH
- Controlling alpha: column type
- “Gotchas” (differences) between HILIC and reversed-phase
Section 8. Gradient separations
- Why/when to use gradients
- Similarities between gradient and isocratic
- Controlling gradients: steepness
- Controlling gradients: range
- Controlling gradients: shape
- Differences between gradient and isocratic: the linear solvent strength model
- Selecting initial gradient conditions
- Dwell volume issues
- Baseline drift & noise issues
Section 9. Quality by Design
- What is QbD
- Combining selectivity parameters
- Semi-automated method development
Section 10: UHPLC-HPLC method transfer
- Equipment considerations: pressure, extra-column volume, detector sensitivity
- Gradient equipment issues: dwell volume, mixing volume
- Scaling parameters
- Method adjustment vs. modification
- “Gotchas”: stationary phase chemistry, temperature control, selectivity/pressure effects, “good column hygiene”
