Teledyne CombiFlash Torrent Manual de usuario

CombiFlash

Torrent™

Quick Start Guide

Overview

These instructions supplement the

CombiFlash Torrent Module Installation Guide (part

number 60-5243-135) and briefly describe the steps nec-

essary to perform a purification run. These quick start

instructions assume that:

●the system has been installed according to sec-

tion 2 of the Installation Guide,

●the operator is familiar with the safety warnings

in section 1 of the Installation Guide, and

●the system will be operated from the touch panel

display.

Quick Start Instructions

1. Perform Thin Layer Chromatography (TLC) on

the sample. From the TLC you'll learn of a suit-

able column media and solvent system, and the

difficulty of the compound separation.

2. With the column media known, select a

RediSep®Rf column size appropriate for your

intended sample load. Refer to the Help topic

“RediSep Rf Column Selection Guide” for recom-

mended sample load ranges. If the separation is

difficult (Rf<0.2), your sample load should be

near the low end of the selected column’s range.

3. Insert the RediSep Rf column on the

CombiFlash Torrent module. PeakTrak software

loads a method for that column. You can manu-

ally select the column media and size if the sys-

tem does not detect the column.

Note

Raising the upper column mount assembly from its

top- or bottom-most point of travel triggers the RFID

column detection. If you exchange a column without

fully lowering or raising the upper column mount,

select the File>New menu command instead to detect

the RediSep Rf column.

4. Ensure that the Solvent A and Solvent B lines are

routed to suitable solvents.

5. Select Tools>Auto Prime to open the Auto Prime

Window (Figure 1). Click the Play button. The

system primes the internal tubing and then

closes the Auto Prime window.

Figure 1: PeakTrak’s Auto Prime window

6. On the main window, click the Play button.

7. From the Run Requirements window, select a

Loading Type.

❍If the sample is soluble in the starting mobile

phase (Solvent A), select Liquid.

❍If the sample is not soluble in the starting

mobile phase, select Solid (Pause).

❍If the sample is loaded directly on the column

media, select None. The separation will begin

immediately without equilibration when you

click the OK button.

Note

Use the Solid option only when the solid sample

cartridge is already prepared and loaded on the

module. The module will equilibrate the column and

then automatically begin the run without user

intervention.

Instruction Sheet 69-5243-134

February 12, 2010

Instruction Sheet 69-5243-134 February 12, 2010

Teledyne Isco, Inc.

P.O. Box 82531, Lincoln, Nebraska, 68501 USA

Toll-free: (800) 775-2965 ˘ Phone: (402) 464-0231 ˘ Fax: (402) 465-3001

E-mail: [email protected]

8. Click OK to start the run.

If you selected Liquid or Solid (Pause), continue with

step 9.

If you selected None, skip steps 9 through 11 and

continue with step 12.

9. The system will equilibrate the column. During

this time, prepare the sample.

❍If you selected Liquid, dissolve the sample in a

minimal volume of Solvent A (or other misci-

ble solvent) and draw the liquid into a

syringe. Alternatively, prepare the optional

Sample Load Pump.

❍If you selected Solid (Pause), prepare the sample

using a solid sample cartridge. Refer to the

Help topic “Preparing Solid Samples” for

instructions.

Figure 2: Pause messages

10. The CombiFlash Torrent module will pause after

performing the column equilibration (Figure 2).

It will ask you to inject the sample or load the

solid sample cartridge.

If Liquid – inject or pump the liquid into the

sample injection port. To force the sample

through the valve, follow this injection with

another 5 mL of the starting mobile phase

(chase solvent).

If Solid – load the cartridge on the

CombiFlash Torrent module. To load the car-

tridge:

a. The solid load cartridge cap must match the

size of your selected cartridge. If it does not

match, install the correct cap now.

b. Press the lever on the side of the cap and fully

extend the plunger.

c. Slide the plunger into the cartridge until it

reaches the top frit.

d. Press the lever and push the cartridge into

the cap. Align the cartridge so that it fits fully

into the recess in the cap.

e. Rotate the cartridge 1/4-turn to secure it in the

cap. For 25 gram cartridges, rotate the car-

tridge clockwise. For cartridge sizes 65 gram

and larger, rotate the cartridge counterclock-

wise.

Note

Refer to the instruction sheet titled “Adjustable Rf

Solid Load Cartridge Cap for RediSep Rf” (available at

www.isco.com) for complete instructions.

11. Click OK to begin the run.

12. Monitor the red absorbance trace.

The CombiFlash Torrent module will proceed

through the run and collect fluids in the fraction

tubes. It isolates peak fluids from non-peak flu-

ids. Tube change marks on the screen and the

tube map help you locate your compounds.

For detailed operating instructions, refer to the

on-line help.

Operation Tip: Difficult Separations —

If the TLC indi-

cates a difficult separation (Rf<0.2), click the Pause

button at onset of each peak in the absorbance trace.

This inserts an isocratic hold and extends the run. The

isocratic hold can increase the resolution by delaying

the release of the next closely eluting peak. When the

absorbance of the peak is close to the baseline again,

click the Pause button to resume the linear gradient

which will help to maintain sharp peaks.

Assistance Available

Should you need assistance with these instructions,

please contact Teledyne Isco.

Liquid Samples

Solid Samples

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