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General Description
FPLC® - fast protein liquid chromatography - is a term created by Pharmacia Biotech (now Amersham Pharmacia Biotech) to describe analyses on their automated liquid chromatography systems that are specially designed to accept Pharmacia columns. An important feature of an Amersham Pharmacia Biotech FPLC automated system is the design of the fittings. Pharmacia® columns and accessories use M6 fittings. These are metric fittings, incompatible with the fittings used in many HPLC or low pressure systems.
Some FPLC columns are shipped with tubing already connected into the ends. Each piece of tubing has a male M6 fitting at its open end, and both of these are screwed into a female-female union to form a closed loop for shipping. You do not want to cut these fittings off the tubing! You must use an adaptor or union to connect the column to alternative systems. Other FPLC columns may be supplied with tubing adaptors that assist in fitting the column to an alternative system.
The following describes the most common scenarios that we encounter in adapting FPLC columns to non-Pharmacia (non-metric) instruments:
Adapting FPLC columns to most HPLC systems:
Option 1: female M6 to male 10-32 adaptor (Cat. No. 55069*)
Most HPLC instruments use 10-32 fittings with 1/16" tubing, so you can use an adaptor (one-piece fitting) to connect the FPLC column to the instrument. The inexpensive approach is an adaptor such as Supelco Cat. No. 55069, a one-piece fitting with female M6 threading at one end, to accept the male M6 fitting on the tubing connected to the FPLC column, and 10-32 male threading on the other end, to fit into your detector or injector. Use of this adaptor will require you to remove the fitting on the injector or detector.
Option 2: union: female M6/male 10-32 fitting to female 10-32/female 10-32 fitting (Cat.No. 55068*)
If you do not want to change the detector or injector fittings on your HPLC, you can use a two-piece union. Again, the inexpensive approach is an Upchurch item, Supelco Cat. No. 55068. The female M6 end of the female/male fitting accepts the male M6 fitting from the FPLC column, the male 10-32 end fits into one end of the female/female 10-32 fitting. The other end of the female/female fitting accepts the 10-32 fitting on your HPLC tubing.
Adapting FPLC columns to non-metric medium/low pressure liquid chromatography systems:
Option 1: female 1/4-28 to male 10-32 adaptor (Cat. No. 55071*) plus 10-32 to female M6 two-piece union (Cat. No. 55068*)
Many medium/low pressure systems use 1/4-28 fittings. You can use an adaptor with a union to connect an FPLC column with M6 fittings to these instruments. This method requires that you currently have tubing with 1/4-28 fittings on your injector and detector. These fittings will thread into the female 1/4-28 to male 10-32 fitting (Cat. No. 55071). The male 10-32 will tread into the two-piece union (Cat. No. 55068) that will then join to the M6 fitting on the Pharmacia tubing.
Option 2: union: female 1/4-28 to female M6 (Cat. No.59259-U*)
Like Option 1, this option requires that the tubing on your injector and detector already have male 1/4-28 fittings. The male 1/4-28 fitting will thread into the union that will then join it to the male M6 fitting on the Pharmacia tubing.
In addition to fittings and empty columns for FPLC and other medium pressure applications, we offer many Amersham Pharmacia Biotech and other ready-to-use columns. For information about these columns, refer to the Products section of our website.
*Be sure to order two of these parts, one for each end of the column.
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