Liquid Chromatography
Separation Science offers free learning from the experts covering methods, applications, eBooks, webinars, eSeminars, expert forums, videos, tutorials and training for liquid chromatography (LC) analysis users.
Featured Educational Resources
Fast and Selective Purification of Oligonucleotides
Video & Application Note
Florian Rieck (Agilent Technologies)
Join Dr Florian Rieck in a short presentation as he introduces a method for the fast and selective purification of oligonucleotides. The complementary application note, which is available for download, describes a purification method with MSD that uses alternative, less costly reagents. Target compounds were collected faster and with higher purity compared to conventional methods.
LC-MS/MS Fundamentals: The Overview
On-demand
by Jack Steed and Jessica Smith (SCIEX)
This presentation discusses the fundamentals of LC-MS technology and how this can be utilized in your everyday workflows. The discussion ranges from the basics of LC separations to the implementation of high-resolution mass spectrometry, including data acquisition and review.
Biphenyl, the Go To Phase for LC-MS Method Development
On-demand
Paul Connolly (Senior Product Manager for Liquid Chromatography, Restek)
Learn about the mechanisms of retention of the Biphenyl phase; best practices for successful Biphenyl utilization in liquid chromatography methods; and see examples of Biphenyl chromatography with applied solutions for multiple market needs. In addition, get an introduction to the ProEZLC Modeller for method development.
Strategies to Enable and Simplify HPLC Polar Compound Separation
On-demand webinar + application and technical notes
Matthew Przybyciel, PhD, R&D Leader (PerkinElmer)
Separation Science, in collaboration with PerkinElmer, offers an on-demand webinar that provides solutions for tackling the most difficult HPLC polar compound analyses using a variety of food and pharma applications. Matthew Przybyciel, PhD, R&D Leader at PerkinElmer will present how, even the most difficult analyses, can be simplified and demonstrate how the latest in column developments can enable easier and faster HPLC analysis of polar compounds for an end-to-end workflow solution that can improve your lab productivity. In addition, the application and technical are also available to download.
[LC Tutorials] How to Improve Method Robustness through Control of Unwanted Interactions
On-demand Tutorial
Kim Haynes (Waters)
This course is designed to take you from beginner to advanced in controlling the interactions within your chromatographic separations. It will cover how to improve method robustness through the prevention of analyte loss, with improved confidence in the quality of your data.
[LC Tutorials] Understanding LC Column Attributes - What's Important for Your Chromatography?
On-demand Tutorial
Jonathan Turner (Waters)
This course is designed to take you from beginner to advanced in understanding column selection and method development. It will cover the key aspects of the LC column from the base particle to the bonded phase, and even the hardware.
HPLC 2022: International Chromatographic Conference
18-23 June 2022
HPLC 2022
Join scientists from around the world at the 50th sequel of the longest-running, largest, and most recognized conference in the world covering all aspects of Separations and Analyses carried out in Liquid Phase. HPLC 2022 will take place on June 18-23, 2022 in San Diego, California, USA. Information about the program, speakers, posters, courses, tutorials, competitions, sponsoring organizations, travel grants, exhibition and more may be found at https://hplc2022.org/
Super Charge your Method Development with a Quick, Easy Universally Compatible LC and LC/MS Method
On-demand webinar
Anne Mack (Application Scientist, Agilent)
This webinar demonstrates how you can achieve enhanced performance for basic analytes under weak ionic strength mobile phase conditions, such as formic acid. A new method development tool is featured: a unique C18 bonded phase chemistry with a charged surface on 2.7 μm superficially porous particles.
Getting your LC/GC Lab Up and Running Again
On-demand webinar
Paul Altiero and Mark Sinnott (Agilent)
Regardless of why your lab may have been shut down, you need to get your LC and GC instruments back up and running quickly and safely. This presentation covers the information you need to get back to your important research including restart procedures, helpful service, items, support topics, and discussions on improving operational efficiency and capacity.
Trials and Tribulations of Fat Soluble Vitamin Analysis by UHPLC-MS/MS
On-demand webinar
Alan Edgington (Biotage, UK)
This webinar summarizes the work performed to develop a method for the analysis of fat-soluble vitamins including vitamins A (Retinol and Beta Carotene), D (25 Hydroxy D3 and 25 Hydroxy D2), E (Alpha Tocopherol) and K (Phylloquinone and Menaquinone) in human serum.
Introduction to and Practical Applications of the SCIEX Triple Quad 7500 LC-MS/MS System – QTRAP Ready
On-demand webinar
Ian Moore and Jianru Stahl-Zeng (SCIEX)
Learn how the SCIEX 7500 System enables you to attain up to 7X greater sensitivity to quantify more trace analytes at lower levels across a broad suite of sample types and workflows.
Making the Most of your LC System
On-demand webinar
Mark Powell (Applications Engineer, Agilent)
This on-demand webinar will cover the LC system overview, from solvent bottles to detector, with a focus on preventive maintenance and what to look out for to keep your systems running at optimal performance.
Ion Chromatography Solutions in the Petrochemical Industry
On-demand webinar
Iris Reber (Metrohm, Herisau, Switzerland) and Ricardo Sales Tena (BP, Castellón, Spain)
In this webinar, you will learn more about IC and discover solutions to some of the challenges petrochemical labs are faced with. Further, you will get to know various inline sample preparation techniques that will allow you to save valuable time and resources.
Comprehensive Two-Dimensional Liquid Chromatography (LC×LC) for the Analysis of Bioactive Compounds in Foodstuffs
On-demand webinar
Luigi Mondello (University of Messina, Italy)
Learn about the benefits provided by comprehensive two-dimensional LC techniques, for the separation/identification of complex food matrices, as well as how to implement LC×LC using different column combinations (e.g. NP×RP, RP×RP and HILIC×RP) and instrumental set-ups.
HPLC Column Selection Guide to Help You Achieve the Best Separation
eBook, Application Notes and Online Tutorials
SHODEX
Learn from the expert, Dr Christian Hirsch from Shodex, who has compiled a comprehensive HPLC column selection guide to help you select the right HPLC column for your application and achieve the best separation out of your HPLC measurements. Apart from the eBook, do check out Shodex's latest application notes and educational online tutorials: Reversed Phase RP, HILIC, Size Exclusion chromatography SEC, Ligand Exchange LEX and Ion Chromatography IC.
Looking for Cost-effective Online Learning in LC?
HPLC Training Courses
Fundamentals of HPLC - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
‘Fundamentals of HPLC’ covers the essentials that every scientist needs in order to make effective use of liquid chromatography instrumentation, including: The basics of the chromatographic process, Terms and definitions of chromatography, How HPLC systems work, Practical tips on preventive maintenance, Principles of separation chemistry, Why “reversed-phase” is the standard separation technique, The meaning of “gradients” and why they are used, How to get the best quantitation and much more. By taking this online course you will acquire a good understanding of the fundamentals of HPLC and get a better knowledge of the key concepts involved. The course is aimed at managers, supervisors, auditors, reviewers, etc. who use information generated by HPLC but do not necessarily do chromatography themselves.
HPLC & UHPLC Troubleshooting - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
'HPLC & UHPLC Troubleshooting: A Performance Qualification Approach' is an intensive online course that teaches you the ins and outs of solving problems that occur with your high-performance liquid chromatography (HPLC) and ultra-high pressure liquid chromatography (UHPLC) methods. You will learn how to qualify the performance of your HPLC or UHPLC using specific tests that also can be used for isolating existing problems. More importantly, you’ll learn how to prevent many of these problems from happening in the first place. This course is designed for anyone who works with HPLC or UHPLC. No previous experience with HPLC or UHPLC systems is assumed; however, much of the course will appeal to the more experienced worker.
Advanced HPLC Method Development using QbD - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
'Advanced HPLC Method Development Using Quality by Design' is an intensive online course that teaches you strategy and technique to develop high-quality HPLC and UHPLC methods quickly and with confidence. You will learn a time-proven technique that walks you through the method development process a step at a time using sound chromatographic principles to help make the process more efficient. The techniques can be used as a stand-alone strategy or added to existing development procedures to help streamline the process. Because the same principles apply to HPLC and UHPLC, you can apply the same information for the development of UHPLC methods. Here’s what the course covers: What Quality by Design (QbD) is and how it applies to HPLC methods?, A review of basic HPLC theory as it applies to method development?, The makeup of HPLC columns including the support material and the bonded phase?, The role of solvents in the separation?, How the column plate number, selectivity, and retention can be used in a logical way to solve even the toughest separation problems?, How to use mobile phase solvents, pH, temperature, chemistry, and other variables to pull apart difficult-to-separate peak pairs?, How gradient elution works and how to remove the mystery of optimizing gradients?, How a scouting gradient can be used to speed the development of both isocratic and gradient methods? How to use a simple spreadsheet calculator to move from the scouting run to the next step in method development?, How computer-assisted method development tools can leverage a small amount of data into a large amount of information about a separation?, How to take advantage of a publicly available internet database to identify equivalent and orthogonal columns?, When standard reversed-phase techniques don’t work, how to choose mixed-mode, ion-pairing, normal phase, HILIC, chiral, or other chromatographic modes?, What’s the same and different when comparing HPLC and UHPLC?, What to look for when validating a method?
Principles of HPLC Validation - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
This class focuses on validating high-performance liquid chromatographic (HPLC) methods developed for the analysis of pharmaceutical drug substance (pure drugs), drug product (formulated drugs), impurities, and degradation products. Although the focus is on pharmaceuticals, the principles apply for most quantitative HPLC methods, such as environmental or general chemical analyses. Here’s what the course covers: What steps are that need to be undertaken during validation?, Why quality is so important?, What basic chromatographic measurements will be used?, Why to approach validation as part of a larger process?, Which regulatory documents are important?, What parameters of the method need to be tested?, What are the different standardization techniques, and when are they used?, What is Quality by Design, and why is it important?, What software tools are available to simplify the process?, What pitfalls should be avoided in validation?, What is pre-validation, and why is it worth my time?, How do I choose a system suitability sample?, How can I tell if the method is out of control?, How much can I adjust a method before I have to re-validate?
HPLC for Biopharmaceuticals - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
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. 'Practical HPLC for Biopharmaceuticals' explains chromatography in practical terms from the ground up. Here’s what you’ll learn in this online course: The basics of isocratic and gradient HPLC, Reversed-phase and ion exchange HPLC, HPLC equipment – from reservoirs/pumps to tubing/injectors to columns and detectors, Understanding and overcoming common column problems, Alternative modes: normal phase, HILIC and size exclusion, Quantitation, UHPLC, Performance qualification, Troubleshooting and diagnostics.
LC-MS/MS for Chromatographers - Training Course
Self-paced online course with validated learning and certificate of completion
Analytical Training Solutions | Separation Science
With the introduction of the mass spectrometer (MS) as a practical detector for high-performance liquid chromatography (LC or HPLC), LC-MS began to be used for routine applications. One form of LC-MS uses a tandem MS (LC-MS/MS), that has become the go-to instrument for quantitative analysis of drugs in biological matrices in the pharmaceutical laboratory. As they have become more widely used and understood, LC-MS and LC-MS/MS have found their way into environmental applications, synthetic chemistry, protein analysis, and many other areas. LC-MS/MS has moved from a technique available only to skilled experts in both HPLC and MS to a routine tool usable much as any other HPLC detector. You will find that this introductory course in LC-MS/MS will give you a good foundation of understanding the technique and how to use it in your daily work. We start out with an overview of different MS techniques and focus the majority of the course on the quadrupole mass spectrometer. You will learn how MS works, how ions are produced and fragmented. Sections focus on qualitative, as well as quantitative analysis. Each part of the process, calibration, tuning, sample preparation, HPLC method development, and MS method development are covered. Many tricks and techniques of practical use are presented.
HPLC Solutions
LC-MS Calibration vs Tuning
Fundamentals of HPLC
HPLC Solutions
There are two procedures that are used to make sure that an HPLC equipped with a mass spectral detector (MS) is reporting proper mass information and that the system is properly set to detect the analyte(s) of interest.
Does a Guard Column Make Sense?
Fundamentals of HPLC
HPLC Solutions
A guard column is one of those HPLC accessories that is a bit of a mixed grill – it can be a good thing or a not-so-good thing. Let’s look at some of the different aspects of guard column use to see if they make sense for you.
Does Air Damage My Column?
Fundamentals of HPLC
HPLC Solutions
I accidentally ran out of solvent in an overnight run on my HPLC system. I had not activated the low-pressure limit on the pump, so the pump just pumped air for half the night. Will this ruin my column? If not, how can I get all the air out?
Pore Size vs Particle Size
Fundamentals of HPLC
HPLC Solutions
What is the relationship between pore size and particle size for an HPLC packing material?
Internal Standards: How Does It Work?
Fundamentals of HPLC
HPLC Solutions
In this article, we’ll look at the use of internal standards in HPLC and LC-MS applications, and in particular the topic of this discussion is how an internal standard is used in quantitative analysis.
What Makes a Good Internal Standard?
Fundamentals of HPLC
HPLC Solutions
In a earlier article we took a brief look at why we might decide to use an internal standard and how a calibration plot is constructed and used in HPLC analysis. This time, we’ll consider some properties that are necessary when choosing a compound to use as an internal standard.