I use an Agilent FCS on a 1200 LC system. It is controlled by the Agilent Chemstation software and works very well and provided your peakwidth is not very wide it should be easy to collect individual peaks into std LC vials, either for further LC analysis or something else. Is suspect other vendors offfer something similar. Cheers Neville

I agree with Neville. if your analytical HPLC-system can perform at high enough flow rates for semi-preparative HPLC, it will be possible to do fractionation. We also use Agilent 1200 systems with fraction collectors, but have set up several other systems with simple fraction collectors as provided by Gilson. If you plan to fractionate from analytical runs, just keep in mind that your sample loading will be very small, leading to very small fractions and long hours of fractionation. Rather invest in semi-preparative columns of the same stationary phase and brand as your analytical columns to cut down on fractionation time,

I do not have experience with FCS for analytical HPLC. What you have to pay attention is to know whether the HPLC software can monitor the FCS.

Indeed, the FCS can work either by peak recognition or by time collection.

In the first way, the software has to detect the peak (minimum slope, minimum height for instance) and send the message to the FCS to start/stop the collection. The problem is that analytical HPLC software are generally ot developped in this way.

The second possibility is to schedule an automatic collection : you will asked to fractionnate every 30 sec for instance ; then you will have to check the composition of the fractions and pool the ones containing your product of interest. By this way, the FCS don't need to be monitored by the HPLC.

So it is feasible but not very convenient.

Hi, There are FCS from Agilent but also from Waters and ISCO (Foxy), and others. The choice of FCS will depend on peakwidth as Neville said, but also on desired compound quantity (mg? g?), column dimensions, flow rate and run time. Some FCS accomodate only 2-4ml vials, other FCS have exchangeable trays with up to 100 or even 200 test tubes (10-20 ml). You can buy sophisticated software to help with peak collection. All FCS can be programed to collect fractions based on time, number of drops or by peak. In this case, the FCS is connected to the LC detector which triggers collection when the peak reaches parameters you can define (slope, height).

Of course you can collect peaks by hand, I've done it a lot in old days...but I'm sure you have better things to do with your time, not to metion that peak collection will not be as reproducible as you might wish, compromising peak purity. Hope this will help you, Luis

I believe a fcs system can be used for fractionation and collection of interested peak for the sake to get some mg amount specially when using analytical columns but the peak shape should be good (not broad) and also with a reasonable tailing facotr.

Dear Buhara, all the above answers are very much to the point. Just to make thinks clear: YES you can isolate compounds using analytical columns as long as you inject as many times as to accummulate enough mass for each fraction, and this have to be done in an automatic manner using software which knows when the peaks come out . Semipreparative columns are usually well within the possibilities of an analytical HPLC but they have higher back pressure and the same analytical conditions may bring up the pressure too much. Also your detector may be out of scale if using semiprep columns and consequently higher amounts of mass overloading the analytical cell.

As all the people above say, to get the most out of your investment go for a fraction collector of the same make of your HPLC and which can be automatically controlled by software. Also check if you have the capability of analysing by MS and NMR the identity of your fractions which will contain tiny amounts of mass (under 1 mg typically after many runs). Said that, I could isolate phenols from Bupleurum very easily and check their identity by NMR (400MHz) injecting 10 times 80 microliters of extract (40mgml) and accummulating the fractions on the same vials (be careful with calculations about how much volume you're collecting and use the proper vials so they do not overflow). The solvent from the vials was evaporated under N2 stream and its residue directly solved in deuterated solvent fro NMR and MS analyses. I was using the same system that Neville describes above, as I just published it in Nat Prod Comm. DO CHECK WITH YOUR HPLC REPRESENTATIVE VERY CAREFULLY TO AVOID SPLASHING THE MONEY IN A FC WHIC IS NOT SUITABLE FOR YOUR PROJECT. THEY USUSALLY WILL ADVISE YOU WELL IN THE BEST MODEL TO SUIT YOUR GOALS.

ithe answer is yes you can use and i would recommend the Agilent technologies system it comes with software. Have a look at this pdf document fromAgilent it may be of help

Hi, I use FCS since several years on HPLC system. I use on Waters system with the Fraction Collector, easy to manage with minimum slope and height and also with time collect fixed on all the chromatogram or by windows. And the Waters FCS can accept eppendorf tubes or 10 ml tubes so you can use 4 mm columns or largers. But for me the negative point is the software interface on Empower to see the fractions on the chromatogram and some time drops fall when the system switching tubes.

The second FCS I use is a Agilent System on a 1290 UHPLC with narrow bore column 2.1mm and I collecte in 96 microwellplates with 50µl to 200µl fraction volume per well. I collecte fractions for Maldi-Tof analysis and I don’t have problems with amount of material. But it’s not the same story with NMR studies (mg).

You can collecte also with eppendorf tubes but only 4x27 tubes. The software Chemstation show you the fraction on the chromatogram and which plate and well position it was collected, quite nice for my point of view.

Hope this will help you, Pascal

As all have stated above yes. Software control is very nice but not absolutely necessary. What is necessary is to know the volume of tubing from detector to collector, or in other words, the delay from detector to collector. You want to be sure of what you collected in the vial. Good luck.

I work for Teledyne ISCO and we manufacture fraction collectors that work with many analytical HPLC systems. They can start based in the injection trigger and collect fractions based on peaks if the HPLC provides an electrical signal.

Our information on the fraction collectors is here:

http://www.isco.com/products/products2.asp?PL=10160&image=Chromatography3.jpg

Here are links to the brochures:

http://www.isco.com/WebProductFiles/Product_Literature/101/Fraction_Collectors/Foxy_R1_and_R2.pdf

http://www.isco.com/WebProductFiles/Product_Literature/101/Fraction_Collectors/Fraction_Collector_Brochure.pdf