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17
How to Use the System
6. Pour the liquid sample or the silica with the adsorbed
sample into the SIM. See Table 4 for maximum volumes.
Note:
When working with oils, dilute the sample with elution
solvent until it will flow freely down a glass rod.
SIM
Max Volume
(mL)
Max Silica Mass
(gram/SIM Unit
*
)
500
350
175
1000
900
450
2000
1800
900
Table 4. Maximum volume and nominal mass of silica for the SIM modules.
*
This is the maximum amount of silica that fits into the SIM; the actual
amount of silica used will depend on the sample being loaded.
7. If loading a solid sample:
a. Tap the silica down using the frit insertion tool.
b. Insert a second top frit into the SIM and push it down
using the frit insertion tool.
Note:
The top frit must be installed or the separation
efficiency will be low.
8. Ensure that the gasket grooves of the lid and SIM body are
free from any particulates.
9. Place the top assembly back in place and tighten the clamp.
Ensure that the sanitary gasket is captured in the grooves of
the lid and SIM body; see Figure 24.
Note:
Use a sanitary gasket that compatible with the used
solvent; see “Sanitary Gaskets” on page 4.
Note:
If the gasket shows any signs of wear or damage,
replace it.
Note:
It is sometimes useful to apply a small amount of
solvent to the groove and gasket to ensure a good seal.
10. To load the sample onto the equilibrated cartridge and elute
fractions, see “Load the Sample onto the Cartridge and
Collect Fractions” below.
Figure 24. The sanitary gasket must be captured in the grooves of the lid
and the SIM body.
Load the Sample onto the Cartridge and Collect Fractions
Collect your fractions in flasks that are grounded or made of glass.
The following are guidelines for vessel types by cartridge volume:
»
Flash 75M (400g): 125-mL flasks
»
Flash 75L (800g): 250-mL flasks
»
Flash 150M (2.5 kg): 1-liter flasks
»
Flash 150L (5.0 kg): 2-liter flasks
Since your compound will not elute in less than one column volume
(CV), you may wish to collect the first 0.5, 1, or 3 to 6 liters in
a larger flask or carboy. As you approach the point where your
compound will elute, use the smaller fraction collection vessels
(see the bullet list above) to maximize your yield and purity.
Note:
Based on the predicted retention volume (CP) of the first
compound, the initial solvent volume could be collected in one
large flask to minimize glassware usage.
Warning
»
Only use waste/fraction collection vessels that are grounded or
made of glass.
11. If the SIM contains a liquid sample:
a. Ensure that the fraction collection nozzle is inserted into
a collection vessel that are grounded or made of glass.
b. Turn the three-way valve on the radial compression
module toward the tube connected to the SIM
c. Turn the
MV-02
valve on the AM-190 manifold to its
on/pressurize position (see Table 2 on page 3) to
drive your sample onto the cartridge.
d. When the sample has been loaded onto the cartridge,
i.e. when the SIM outlet tubing is dry, turn the
MV-02
valve on the AM-190 manifold to its off/vent position
(see Table 2 on page 3).
12. Turn the three-way valve on the SIM toward the tube
connected to the solvent reservoir.
13. Ensure that the solvent reservoir is pressurized, i.e. the
MV-03
valve on the AM-190 manifold is turned to its
on/pressurize position (see Table 2 on page 3).
14. Set the solvent flow rate/solvent pressure by adjusting the
PRR-02
regulator on the AM-190 manifold as follows:
»
Flash 75M: approx. 40 psi (2.8 bar)
»
Flash 75L and Flash 150M: approx. 60 psi (4.1 bar)
»
Flash 150L: approx. 60 to 80 psi (4.1 to 5.5 bar)
Note:
These solvent pressure settings are approximate
guidelines. Your particular mobile phase, depending on its
viscosity, may require a higher or lower pressure. For flash
cartridges packed with KP-Sil silica, Biotage recommends a
solvent flow rate between 100 and 250 mL/min for Flash 75
and between 400 and 1000 mL/min for Flash 150.
Note:
Ensure that the solvent pressure is at least 20 psi
(1.4 bar) lower than the radial compression pressure.
Summary of Contents for Flash 150
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Page 34: ...32 Biotage Flash 75 150 User Manual Biotage 2018 Notes Notes ...
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