1. ITSP Setup on a
PAL-xt PAL System
Autosampler
SmartSPE™
2. ITSP Hardware Kit
2
ITSP Prep / Waste Station
(ITSP Cartridge Tray) ITSP PAL Lower Needle Guide
The crown of the ITSP cartridge seal mates
with the nipple on the ITSP PAL Lower Needle
Guide to center the needle in the cartridge
ITSP Setup on PAL-xt
SmartSPE™
3. ITSP Cartridge Trays
and Waste Stations
ITSP Hardware Kit
3ITSP Setup on PAL-xt
ITSP-modified PAL
Lower Needle Guide
DLW2 option
preferred
SmartSPE™
8. ITSP Cartridge Disposal Location
Vial “ITSPdisp” - the location to dispose of
spent ITSP cartridges – is most often
programmed to be located off the front end
of the wash station.
8ITSP Setup on PAL-xt
ITSPdisp
vial location
SmartSPE™
9. ITSP is a Consumable Solution, Not an Instrument
ITSP Setup on PAL-xt 9
ITSP (Instrument Top Sample Prep)
is chromatographic SPE
The heart of the ITSP system is a patented cartridge containing
customer-defined filter and/or packed chromatographic media.
Using a PAL System autosampler, the ITSP cartridge can be
moved over trays or vials to totally automate sample clean-up.
With ITSP, you get LC performance in a disposable SPE cartridge!
SmartSPE™
10. ITSP Cartridge Components
10
PAL Lower Needle Guide
(modified for ITSP)
PAL Syringe Needle
SPE Sorbent or Filter Media
Small inner diameter of ITSP needle guide
- reduces volume to improve efficiency
- maintains vertical position of ITSP
cartridge during transport.
Needle Guide
Septum grips needle
to allow the PAL to transport
the ITSP cartridge.
Septum
Media Cup
8mm Crimp Seal
The crown of the ITSP cartridge seal
mates with the nipple on the ITSP
Lower Needle Guide to center the
needle in the cartridge.
ITSP Setup on PAL-xt
SmartSPE™
11. A funnel cone beneath
the septa guides the
syringe needle into the
bore of the cartridge.
A dispersion cone
beneath the syringe
needle tip disperses
the solvent/sample
over the sorbent bed.
The needle penetration for
ITSP tray types is set by
default to 20mm.
• This allows the tip of the needle
to be positioned at the top of the
dispersion cone above the sorbent
bed.
• Longer penetration may cause the
needle to core the sorbent bed.
• Shorter penetration may not keep
the cartridge aligned properly in a
vertical orientation during
transport.
20mm
ITSP Cartridge Details
11ITSP Setup on PAL-xt
SmartSPE™
12. The analytical syringe replaces the
column sample reservoir above the
sorbent bed as found in manual SPE
cartridge formats
The needle penetrates the septum and
creates a seal so that the sample or
solvent is forced through the sorbent
bed by positive liquid pressure when
the syringe plunger is depressed
Sample loading and elution is
performed at the optimum velocity
(flow) for the chromatographic
separation type
The sample is eluted into a collection
plate or vial and then injected into the
LC injection valve for online SPE-
LC/MS/MS
Unlike other cartridge, plate, tube, or
pipette tip SPE formats which require a
vacuum manifold or pneumatic pressure
(at typically 1 drop/s or 50 L/s +/-20%),
ITSP solvent and sample flow rates can
be precisely controlled at a resolution of
10 nL/s with the PAL System syringe.
(ITSP flow rate is over 1,000 times more
precise).
How ITSP Performs SPE
12ITSP Setup on PAL-xt
SmartSPE™
13. Important ITSP Concepts
• It is very important to the successful performance of an ITSP PAL that tray objects, especially the ITSP
cartridge tray (ITSPprp#), be squarely and securely (to the point that the waste block cannot move at all)
installed and then also very carefully trained after installation. While this is an important point, minor
errors in setup and position training, or minor slippage of trays during operation, can be compensated
for by the ITSP crown-nipple mating action. The mating of the crown on the top of the cartridge seal
with the nipple on the bottom of the ITSP PAL Lower Needle Guide as the needle approaches cartridge
compensates for minor misalignment factors.
• However, the ITSP crown-nipple mating action can only compensate for one or two millimeters of error
in the X or Y axes. In addition, for ITSP transport, the PAL Lower Needle Guide is blocked; so the PAL
cannot use its Z-axis ‘seek object’ function when moving to a tray position while transporting a cartridge.
• When training Reference Positions for the ITSP trays adjust the Z-position such that the lower needle
guide just touches the top of the ITSP cartridge, and then back off (up) one half millimeter (0.5mm).
• An ITSP cartridge should be held in position above the tray when training the Reference Position for the
ITSP elution tray (ITSPelt#).
• Septa in the seal of the ITSP cartridge are thick in order to provide a strong seal against the needle for
reliable transport and for pressurization. The needle choice for the PAL system must consider the septa,
solvents being used, and desired flow rate range, as any choice in one of these can affect the others. We
are happy to make suggestions on ways to achieve balance in each/all of these choices based on your
goals.
• ITSP is chromatographic SPE. Optimal method outcomes require flow rate optimization experiments
(flow vs. dispersion [recovery]) to be performed (just like LC) to achieve full chromatographic
performance.
13ITSP Setup on PAL-xt
SmartSPE™
