Translator for HPLC HINTS and TIPS for Chromatographers

Saturday, April 21, 2018

The HPLC Restriction Capillary; Troubleshooting, Qualification and Running Without A Column:

When we want to closely replicate the operation of an HPLC system under "normal" conditions and do not want to use an HPLC column in-line, we install a "restrictor" in its place. A restriction capillary is often a very narrow ID section of long tubing (capillary) which will restrict the flow of mobile phase through it. For most HPLC systems, a restrictor which provides about 1,000 psi (~ 70 Bars) of back-pressure will closely replicate normal operating conditions. The restrictor can be chosen based on length, ID, volume and your flow rate to create this level of back-pressure. You could place a high pressure rated, zero-dead-volume union its place, but in doing so, the system back-pressure may be extremely low and show poor pump performance. An HPLC column, with its densely packed small particles inside adds a great deal of back-pressure to the HPLC system and that back-pressure greatly improves the stability of the baseline. HPLC Columns prevents pulsations by acting as a dampener and/or system buffer. Most types of HPLC pumps will not operate properly without 20 or more bars of back-pressure on their outlets to prevent cavitation and excessive pulsation. Columns play a vital role in stabilizing the baseline during an analylsis.

There will be times when you need to operate the HPLC system without an HPLC column installed.

For Example: 
  • Troubleshooting sources of contamination, carryover or artifact peaks on a column;
  • Measuring the HPLC system delay volume (gradient delay);
  • Testing the performance of the injector;
  • Testing the performance of the pump; 
  • Testing the performance of a detector module;
  • Running HPLC Operational Qualification Tests (OQ);
  • Running HPLC Installation Qualification Tests (IQ);
  • Running Performance Verification Tests on a Module (PV);
  • Running many of the ASTM Tests (e.g. "Baseline Noise & Drift Test").
Example of a commercially available Restriction Capillary (Agilent P/N G1312-67500). You will want to include any needed details of the restriction capillary chosen for your work in the SOP's that you write which utilize it as part of a test (P/N, source, volume...).

2 comments:

  1. Great Article!! Now I understand why we need to create instrument test methods which isolate the module by removing the column from the system. Tried to find answer on chromacademy site and received lots of terrible advice from what appeared to be sales people and novice users (they have really bad authors), but at this site, knew we would find the best information.

    PLEAS KEEP up these posts. Our lab in the UK has learned SO MUCH from you.

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    1. Happy to learn you appreciate these scientific posts. Thank you.

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