Operating an HPLC (High-Performance Liquid Chromatography) system can be overwhelming for a new user. There are numerous pitfalls or “gotchas.” Even the matter of which tubing and fittings are compatible, and which are not, can pose a serious impediment to successfully operating the HPLC.
In this discussion, we will cover:
- The significance of connections
- Considerations for the mobile phase
- Quality considerations
- Preventing microbial growth
- Compatibility of sample and solvent
- Characteristics of the column; their significance and limitations
- The importance of sample preparation
Agilent Technologies, Inc.
Rita Steed began supporting the LC column line for Agilent in 1999. As a Chromatography Specialist (first with Chromatography, Inc., then directly for Agilent), Rita worked on-site with researchers in Pharmaceutical and other industries presenting technical seminars and assisting researchers with troubleshooting and method development. Rita has over 20 years of Chromatography experience in the Biotechnology, Chemical, and Pharmaceutical industries. She has held positions in Research, Sales, and Technical Service. Rita has earned degrees in Microbiology and Life Sciences/Biochemistry. In her current position, she is an inside Application Engineer supporting LC columns.
Technology improvements in liquid chromatography/mass spectrometry have enhanced the detection and identification of metabolites and lipids from complex biological samples. As metabolomics and lipidomics measurements become increasingly valued, there is a growing need to automate sample preparation workflows.
Specifically, Agilent automation offers intuitive workflows that provide high data reproducibility and increased throughput while reducing hands-on time. In this webinar, we describe key learnings revealed during the automation of several workflows that extract metabolites and/or lipids from plasma and mammalian cell samples.
Genevieve Van de Bittner, Ph.D.
Agilent Research Laboratories
Agilent Technologies, Inc.
The success of any analytical laboratory technique, lies first and formemost with the integrity of the sample and secondly with sample Preparation. Learn the fundamentals and solutions Chemetrix provised to ensure analytical success
Chromatograpahy & Mass Spec
Since most samples encountered in a chromatography laboratory are not in a form to be directly placed into the analytical instrument, some form of preparation is required for nearly every sample. The sample preparation could be as simple as “dilute and shoot” or as complex as multistage sample handling. The aim of sample preparation is to provide a sample aliquot that (a) is relatively free of interferences, (b) will not damage the column or instrument and (c) is compatible with the intended analytical method. In chromatography, the sample solvent should dissolve in the HPLC mobile phase or be injectable into a GC column without affecting sample retention or resolution, the stationary phase itself, and without interfering with detection.
Here you can find useful resources on Sample Preparation for Chromatography Techniques
Sample Preparation Fundamentals for Chromatography
Simple Approaches to SPE Method Development
Simple Steps to Maximize Lipid Removal with EMR-Lipid