ITSP Setup on PAL-xt

ITSP Setup on aPAL-xt PAL

System Autosampler

SmartSPE™

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-xtSmartSPE™

ITSP Cartridge Traysand Waste Stations

ITSP Hardware Kit

3ITSP Setup on PAL-xt

ITSP-modified PALLower Needle Guide

DLW2 option preferred

SmartSPE™

Solvent Reservoirs

ITSP PAL Accessories

4ITSP Setup on PAL-xtSmartSPE™

Tray HoldersCTC# PAL MTHolder



SmartSPE™

ITSP Elution Trays



SmartSPE™

Sample Trays



SmartSPE™

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.


ITSPdispvial location

SmartSPE™

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™

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 needleto allow the PAL to transport the ITSP cartridge.

Septum

Media Cup

8mm Crimp SealThe 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-xtSmartSPE™

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


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


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.


Documents

ITSP Setup on PAL-xt