How can I improve my HPLC?

How to Improve Resolution in HPLC

Increasing column length.
Decreasing particle size.
Reducing peak tailing.
Increasing temperature.
Reducing system extra-column volume.

How do I make my HPLC peaks sharper?

If this is the case, injecting a smaller amount of sample should result in sharper peaks. Reducing the sample size by a factor of 10 and observing the peak shape is a quick way of determining whether you are operating in the linear range of the detector.

What makes HPLC high performance?

HPLC allows for much smaller particles for column packing materials. This means there is a greater surface area for the stationary phase and the components flowing past it, creating better separation. Better separation means better results and easier analysis, making HPLC a highly recommended separation technique.

What is HPLC troubleshooting?

In an HPLC system, problems can arise from many sources. First define the problem, then isolate the source. Use Table 1 to determine which component(s) may be causing the trouble. A process of elimination will usually enable you to pinpoint the specific cause and correct the problem.

How do I stop fronting in HPLC?

Volume overloading-Injecting too large of a volume can result in fronting, since it broadens the peak. You can eliminate this possibility by injecting a smaller volume.

How do I start HPLC?

For setting up the HPLC machine:

Make sure you have all your buffers set up.
Open the purge valve and purge the system for 5 minutes.
Add your samples into the autosampler tray.
Stop the purge.
Close the purge valve.
Run the system at a normal flow rate (1 ml/min) with your buffer to equilibrate the column for 10 minutes.

What are the disadvantages of HPLC?

What Are the Disadvantages of HPLC?

Coelution. Because of the speed of HPLC and it reliance on different polarities of compounds, two compounds with similar structure and polarities can exit the chromatography apparatus at the same time or nearly the same time.
Adsorbed Compounds.
Cost.
Complexity.

What can HPLC detect?

HPLC has the ability to separate, and identify compounds that are present in any sample that can be dissolved in a liquid in trace concentrations as low as parts per trillion.

What are the tips and tricks for HPLC and UHPLC?

HPLC Tips and Tricks Tiziana Ladisa Sales Support Specialist for Chromatography Italy Thermo Fisher Scientific, Rodano (MI) 2 Overview Thermo Scientific™ Vanquish™ UHPLC system Thermo Scientific™ UltiMate™ 3000 UHPLC system 3 Common Recommendations Mobile Phase 4 Common Problems: Mobile Phase •Solvent compatibility

What should I do before using a HPLC pump?

Before using an HPLC system, you should prime all of the lines in your HPLC pump. This is needed to purge any air from the tubing, introduce fresh mobile phase to each line and then to VERIFY that each channel delivers the reported amount of fluid (measure it). This is a LONG, detailed article with lots of information, Hints and Tips.

How to avoid ” avoiding beginner pitfalls ” in HPLC?

4/24/2013 Avoiding Beginner Pitfalls Agilent Restricted Extra Column Volume Default 1290, 8.6 µL Extra-column Volume Default 1290, 8.6 µL Extra-column Volume Optimized 1290, 3.0 µL Extra-column Volume P=227 bar Rs 5,6 =2.38 N 4 =5529 N 8 =9697 N 9 =9947 P=230 bar Rs 5,6 =2.77 16% increase N 4 =8864 60% increase N 8 =11251 16% increase N

What should the draw speed be for HPLC?

(normal HPLC eluents and samples dissolved in starting eluent) => 10 µL injection volume -> recommended draw speed: 2 – 3 µL/sec => 2 µL injection volume -> recommended draw speed: 0.2 – 0.3 µL/sec ● Draw speed has to be adapted in case of samples and eluents with higher viscosity.

How can I improve my HPLC? How to Improve Resolution in HPLC Increasing column length. Decreasing particle size. Reducing peak tailing. Increasing temperature. Reducing system extra-column volume. How do I make my HPLC peaks sharper? If this is the case, injecting a smaller amount of sample should result in sharper peaks. Reducing the sample size…