When should I replace my LC column?

When should I replace my LC column?

If the inlet is irreversibly plugged or the stationary phase fouled with contaminants you will need to replace column.

How do you regenerate HPLC columns?

How to regenerate the C18/C8 HPLC columns?

  1. Disconnect the column from the detector.
  2. Rinse with a mobile phase without buffer salts (water/organic phase) firstly.
  3. Then rinse with 100% organic phase (methanol and acetonitrile).
  4. Check that the pressure returns to normal, and if not, proceed to the next step.

What is regeneration of column?

Column regeneration. Exposure of a column to samples or solvents containing highly adsorptive components will result in increased back- pressure and a change in selectivity. Often the column can be restored to original performance by suitable wash protocols.

How long does a LC column last?

AB: A LC column can be fine for a long period of time and has no defined shelf life, but it is dependent on what storage solvent was used for column storage. If the column was properly flushed after use and stored in 100% organic solvent such as acetonitrile, the column will survive for many years.

How long do chromatography columns last?

A related question that we hear quite often is “what is the shelf life of a capillary GC column.” Once again, there are no right or wrong answers and GC columns usually don’t ship with an expiration date on them – but in general, a pretty good guideline guide line is to use the column within about two years to ensure …

How do you regenerate HPLC C18 column?

How To Regenerate a C18 HPLC Column

  1. Disconnect column from detector and run wash solvents into a beaker.
  2. Start with your mobile phase without buffer salts (water/organic).
  3. Next, use 100% organic (methanol or acetonitrile).
  4. Check pressure to see if it has returned to normal; if not, then:

How do I clean my new HPLC column?


  1. Change the mobile phase to filtered distilled water.
  2. Allow the water to flow through the column at the rate of 1ml / min.
  3. Change the mobile phase from water to HPLC grade 80% Acetonitrile.
  4. Set the instrument to flow rate 1 ml/ minute and wash the column for 30 minutes.

Do HPLC columns expire?

It should also be noted that good chemical practice dictates that chemicals (including columns) stored beyond three to five years should be re-evaluated for use or replacement before continued use.

How long is an HPLC column good for?

Normal phase column have slightly shorter life spans, but should accept about 1000 injections. When subjected to extreme conditions, most columns will be stable for only a few hundred injections or less.

How often should you change a GC column?

Getting Started

Topic Frequency
GC Septa, GC (non-MS) Every 100 injections or every 3 days
Inlet Liners Inspect daily
Liner O-Rings Every 5 liners
Inlet Seals Every column change or every 6 months

How do I remove air from HPLC column?

Place a thumb over the sealed column top and invert the column until the bubble is in the exit tip. 4. With your thumb, apply gentle pressure to the “diaphragm” created by the laboratory film until the trapped air is expelled from the tip.

Can we wash HPLC column with hot water?

If you then remove your column from the system containing 100% organic, this means you already washed the rest of the buffer but you can use hot water e.g. 60C (but with gradual switching from 100% organic) without problem.

How do you clean a BEH c18 column?

Purge the pumping system of any buffer-containing mobile phases and connect the inlet end of the column to the injector outlet. 2. Flush column with 100% organic mobile phase (methanol or acetonitrile) by setting the pump flow rate to 0.1 mL/min and increase the flow rate to 0.5 mL/min over 5 minutes.

How do you unclog an LC column?

First, wash the column with varying solvents at a low flow rate. However, if your column is so clogged that it can’t flow at all, flip the column and run it in the opposite direction. If all else fails, then you might need a new column.

How do you prolong the life of an HPLC column?

How can I prolong column lifetime?

  1. Sample preparation. Prefiltering samples prior to injection onto the column can help to limit the addition of particulates to the system.
  2. Mobile phase. The mobile phase is another factor that influences column lifetime.
  3. In-line filters and guard columns.
  4. Maintenance.

What is a buffer mobile phase?

As many of you know, buffers are usually used in mobile phases to hold the pH constant. They are composed of a weak acid or weak base, combined with its conjugate base or acid in solution that is at least partially aqueous (at least 50%).

What should I do with my LC columns after column regeneration?

After column regeneration, it is important to store your LC columns with end plugs securely fastened and with your column manufacturer’s recommended solvent for either short- or long-term storage. If you enjoy these videos, please like, share, and subscribe.

What happens to the column after a separation?

We usually make the assumption that, after a separation, all the material initially present in the column or cartridge has been eluted. After a run, the column is simply washed with 2-3 column volumes of the initial solvent mixture before starting a new separation.

What is siliachromcleaning and regeneration?

SiliaChromCleaning and Regeneration Procedures If adequate care is taken, it is possible to maintain column efficiency and reliability over an extended period of time. This section is intended to give information on the different procedures to help extend HPLC column lifetime. Difference between cleaning and regeneration

Should I use a reversed flow during column cleaning?

Strongly adsorbed species are collected at the solvent-inlet end of the column and in many cases it is a benefit to use a reversed flow during washing. Contrary to popular belief reversing the flow will never harm a column that is well packed.