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Frequently Asked Questions

What type of tubing/capillaries and connectors should be used?

Capillary tubing

When working with analytical HPLC instruments, capillaries with external diameter of 1/16" (1.6 mm) are generally used to connect the different parts of the instrument. Those parts that do not affect the extra column volume should have internal diameters (I.D.) of 1 mm. They bring along little risk for clogging and show minimum flow resistance, which contributes to the extra column pressure drop. For those connections critical for the extra column volume, internal diameter of 0.17 mm is recommended. Such capillaries add only 23 μL extra volume per meter of tubing (1.0 mm ID tubing: 800 μL/m). In order to avoid clogging of such fine capillaries, it is of uppermost importance to filter both, mobile phases and sample solutions.

For preparative columns the tubing has to be scaled up in order to reduce the flow resistance caused by the higher flow rates (δPcap≈dcap-4):

Tubing sizes for column sizes
Column I.D.
Tubing I.D.
20 0.25
50 0.5
100 1.0

Fittings are connecting the various parts of the HPLC instrument. Steel fitting usually have a separate ferrule, some designs even use two ferrules, a ring and a conical piece. When tightened the first time, the ferrule is pinched permanently and the surrounded capillary is slightly compressed. The ferrule can no longer be removed from capillary. Plastic (PEEK) fittings are one single piece (nut, ferrule), and are installed by finger tightening only. Over tightening (common with steel fittings) must be avoided, since excessive force is likely to damage the threading and the capillary. It is recommended to tighten the screw only slightly with a wrench and to test if the connection is pressure resistance, meaning, no leakage occurs once mobile phase is pumped through the system.

The distance from end of tubing to ferrule lowermost end is depends upon the column and/or instrument design. Some examples are given below:

  • 0.130" in case of Waters columns
  • 0.80" for Valco design
  • 0.170" for Rheodyne injector design