|To find out
more about customizing this course to meet your specific needs, give us
a call at
or email us
||Virtual On-Site: Practical HPLC for Biopharmaceuticals
should take this course?
If you are a biochemist or biologist with limited experience using
HPLC, this course will acquaint you with instrumental techniques for
analysis and lab-scale purification of both biopolymers and small molecules. If you are an
analytical chemist with experience in the use of HPLC separations for
small molecules, this course will help you apply these techniques to
peptides, proteins and nucleic acids.
does it cover?
Although “small molecule” and “biopolymer” separations have
traditionally been considered as separate activities, analysts in the
biopharmaceutical industry regularly have to deal with both.
Fortunately, the underlying principles of chromatography apply equally
well in both situations when interpreted appropriately.
In this course you will learn:
You will learn how to chose the appropriate separation technique for
your application and how to select the best chromatographic mode to meet your separation goals. You will
learn how to develop and optimize an LC method and how to troubleshoot
it when problems occur. You will learn how to handle columns to achieve
good performance and satisfactory lifetime. The focus of this course
will be on practical guides and tips to achieve successful biomolecule
of HPLC mechanisms of band broadening for both large and small molecules
overview of HPLC instrumentation with an emphasis on preventive
maintenance and troubleshooting
and mobile phase selection for reversed-phase, ion-exchange and
for developing and optimizing HPLC methods
column maintenance and troubleshooting
When is the course available?
course is presented for groups of 5 - 30 people using Citrix GoToMeeting service. Virtual On-Site is six 2.5-hour
sessions (typically 2 or 3 sessions per week). The usual lead time to
schedule a course is approximately 4-6 weeks.
us at (925) 297-5374
or email to firstname.lastname@example.org to arrange timing,
pricing, and content.
What topics are covered?
course content can be tailored to your specific needs. A typical
schedule looks like this:
Section 1. Isocratic HPLC Basics
- What is HPLC?
- Separation Chemistry
- Reversed phase
- HILIC (normal phase)
- Ion exchange
- Ion pair
- Size Exclusion (GFC & GPC)
- Key Measurements: Retention
- Key Measurements: Selectivity, Efficiency, Resolution, & Tailing
- Controlling Resolution
Section 2. Gradient HPLC Basics
- Why use a gradient?
- Gradient is Just Like Isocratic
- Gradient is Different From Isocratic
- Gradient Hardware Issues
Section 3. Reversed-phase HPLC
- Small Molecules: Solvent Selection
- Small Molecules: Column Selection
- Small Molecules: pH, Additives, & Temperature
- Biopolymers: Solvent & Column Selection
- Biopolymers: Temperature, Denaturation, & Additives
Section 4. Ion-exchange HPLC
- General Mechanism & Packings
- Proteins: Gradients, pH, & Particle Size
- Oligos, Peptides, & Sugars (Oh My!)
Section 5. HILIC (Hydrophilic Intraction Chromatography)
- What is HILIC (and what is it good for)?
- How Does HILIC Work?
Section 6. Size-Exclusion Chromatography (GFC and GPC)
- How Does SEC Work?
- Controlling SEC
Section 7. HIC (Hydrophobic Interaction Chromatography)
Section 8. HPLC Equipment: Reservoirs & Pumps
- Reservoirs and Degassing
Section 9. HPLC Equipment: Tubing & Injectors
Section 10. HPLC Equipment: Columns
- Overview & Physical Problems
- Non-Column Problems (I)
- Non-Column Problems (II)
- Chemical Problems
Section 11. HPLC Equipment: Detectors
- UV Detectors
- Gradient Baseline Issues
- Other Detectors
Section 12. Quantitation
- Measuring Peak Area
- Calibration Issues
- Reproducibility Issues
Section 13. Performance Qualification
- Pump & Detector Checks
- On-line Mixing Checks
- Chromatographic Checks
Section 14. Troubleshooting & Diagnostics
- Preventive Maintenance, Documentation, and Troubleshooting
- A Troubleshooting Checklist