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HPLC: install capillaries

Basic questions from students; resources for projects and reports.

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Sorry, I'm a HPLC newbie!

- When I've put a ferrule and screw on the capillary, can I just draw it in the port where it should be used? Is the ferrule then fixed on the capillary or are there any tools necessary?

- Is it common to cut capillaries by oneself or are precut capillaries normally used?

Thanks in advance! :D
Chroma:

Unfortunately, I don't think you've provided enough information for me to answer your specific questions, but here are some general comments.

The term "capillary" simply means narrow tubing. In HPLC capillary tubing usually refers to one of several things:
  • fused silica
    1/32 inch outside diameter (OD) (or less) hard polymer tubing (usually PEEK)
    1/32 inch OD metal tubing (usually stainless steel)
    PEEK-clad fused silica
Each of these tubing materials requires its own types of fittings and cutting procedures.

Fused silica can be cut by hand in the lab using various types of inexpensive diamond scribes. (New Objective sells a very simple one http://www.newobjective.com/products/ess_tools.html.) For micro and nanoflow systems, it's a good idea to inspect the cuts with a low-power microscope or good hand lens to insure the cut is clean.

Capillary PEEK is more difficult to cut, but IDEX Health and Science (http://www.idex-hs.com/default.aspx) sells such a cutter.

Although 1/16 inch OD and bigger steel tubing can be cut by hand, the results are poor. It's hard to get a straight, square cut. I've never tried to cut 1/32 inch. Most people use pre-cut steel with polished ends.

PEEK-clad silica can not be cut at all and is only used in pre-cut lengths.

As far as fittings go, I highly recommend finding a copy of the book "All About Fittings" by John W. Batts IV. This book used to be distributed for free by Upchurch but seems to have disappeared since Upchurch was acquired by IDEX. The IDEX fittings catalog also makes excellent reading.

Most fittings consist of a ferrule and compression nut ("two piece") or a combined ferrule/compression nut ("one piece"). There are also some fittings that use two ferrules. Some capillary tubing fittings also require a "sleeve". For these fittings, the ferrule compresses on the sleeve and the sleeve compresses onto the capillary. Some compression nuts just need to be "finger tight" while others require some sort of wrench.

The best way to assemble a fitting (usually) the following:
  • Slide the nut and ferrule way up the piece of tubing.
    Press the end of the tubing into the port so that the bottom of the tubing makes contact with the bottom of the port.
    While keeping the tubing pressed into the port, slide the ferrule and nut down the tubing and tighten. This step is important because it insures that there will not be a gap between the end of your tubing and the bottom of the port.
I hope that helps!

If you are talking about cutting capillaries, then you probably mean about naked silica capillaries and in most cases you will need a sleeve (or an appropriate internal ID piece of PEEK) to do what I think you are trying to do with some exceptions...

Making connections like that is a little bit of art and it would be great if someone senior in your organization can show you how to do it...

You did say you were an "HPLC Newbie" so I assume you are talking 1/16" OD capillary tubing.

Stainless steel (SS) tubing can be cut, but you need special tools. A wire-cutter is NOT that special tool....

If I have to use SS tubing, I usually use a pre-cut length.

Mostly I like to use PEEK tubing--easier to cut, but you still need a special tool.

What nobody else has said, so I will, is that you need to pay attention to the ID of the capillary tubing.

You also need to match ferrules to their use, especially the SS ones. Some are interchangeable, but others are not and will create pockets or gaps if used incorrectly.

Once an SS ferrule is properly swaged to an SS tubing, it's there forever. PEEK ferrules and tubing can sometimes be reused, and are also more forgiving when it comes to matching.

The old Upchurch book was great. The new VICI catalog has lots of great diagrams, but doesn't show the effect of mis-matches. There is a nice diagram on p 344 of the 2007 Restek catalog showing incompatible fittings.
All standard disclaimers apply. My posts are my opinions only and do not necessarily reflect the policies of my employer.

"Baffled" Chromatographer, Kostas Petritis and MaryCarson: Thank you for your replies! :)

I meant stainless steel tubings. I believe they have 0,25mm ID. There's also a tool for cutting. Thanks for your detailed description, Baffled Chromatographer. Assembling worked out that way.

I'll try to get the book and catalog about fittings. What I'm furthermore urgently looking for: a book about troubleshooting and mainting a hplc system (for newbies).


- Chroma

A good introductory book on HPLC in general is Mike Dong's Modern HPLC for Practicing Scientists

For troubleshooting, Dolan & Snyder's Troubleshooting LC Systems is old, but still relevant; it has lots of diagnostic flow charts.
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374

I've accidentally found (and ordered) the latter yesterday. I'm glad you also recommend it. If anyone else knows any other "primers" just mention them. Thank you.

- Chroma

A good introductory book on HPLC in general is Mike Dong's Modern HPLC for the Practicing Scientist...
I've been doing HPLC for less than 2 years, so I'm a newbie too - I reviewed this book on the board here. It's in the book review section.
Good Luck!
Kind Regards,
Jade Barker

A good introductory book on HPLC in general is Mike Dong's Modern HPLC for the Practicing Scientist...
I've been doing HPLC for less than 2 years, so I'm a newbie too - I reviewed this book on the board here. It's in the book review section.
Good Luck!
The TOCs is available at amazon. Looks quite interesting, although the main part seems to be rather method development than troubleshooting. I ordered it through Interlibrary-Loan.
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