ITSP Dual Rail Project
- 1. ITSP/LC/MS/MS Opiates Method Customized for High Throughput on a Dual Rail PAL ITSP Solutions
- 2. AGENDA • Introductions • ITSP Overview • Review Proposed Modifications • Open Discussion • Implement Proposed Modifications • Test-run ITSP/LC/MS/MS Opiates Method • Training • Discuss Next Steps
- 3. ITSP Team • Kim Gamble – President of ITSP Solutions • Thurmond Allsup – Applications Chemist at OpAns • Rick Youngblood – Technical Director at ITSP Solutions
- 4. Project Objectives • To propose a solution for a completely automated opiates method which: • has a low capital cost, • reduces sample cycle time to increase throughput capacity without adding workstations, • improves chromatography results through better sample preparation over dilute and shoot – SPE concentrates analytes of interest – SPE eliminates interference compounds, • greatly reduces maintenance on the LC/MSMS (5 to 20 times), • extends the useful life of existing capital equipment (legacy PAL and Quantum Access), • allows reallocation of staff performing manual sample prep, and • is fully automated in a robust manner.
- 5. ITSP Cartridge Autosampler syringe needle Crimp seal (ITSP “crown”) Rubber/Teflon septum Needle Guide Component SPE sorbent Media Cup Component PAL Lower Needle Guide (modified for ITSP)
- 6. ITSP PAL Lower Needle Guide The nipple on the bottom of the ITSP LNG mates with the crown on ITSP cartridge seal
- 7. ITSP Prep Station 96 well custom plate to hold and prep ITSP cartridges Custom waste receptacle plumbed through MT Holder
- 8. 07-UBCXP10-20A • ITSP SPE cartridge • United Chemical Technologies (UCT) Styre Screen® BCX sorbent • Polymeric support functionalized with Benzenesulfonic Acid (Strong Cation eXchange) • 40-63 micron particle size • 100Å pore size • 10mg bedmass • Filter membrane above sorbent bed • 10micron PE frit below sorbent bed
- 9. Proposed Hardware Modifications • Move the Cold Stack 40mm to the right. • Remove the valve assembly from the left rail to make room for ITSP tray holders. • Add a second Fast Wash Station on the left rail for SPE solvents. • Add two MT Holders on the left rail for ITSP prep stations. • Add ITSP drawer pulls on the left side of the Cold Stack. • Replace the left injection unit lower needle guide with the ITSP Lower Needle Guide. • Connect a custom synchronization cable between the rails. • Connect 2 of the valves on the right rail to alternate between 2 LC’s feeding the MS.
- 10. Dual Rail PAL Synchronization Cable Interface 2 Interface 2 DB25M DB25M SYNC_SIG: LCINStrt Active state: High TTL-In1 1 4 TTL-Out1 Active State: High OUT_SIG: LCINStrt SYNC_SIG: LCINStop Active state: High TTL-In2 2 5 TTL-Out2 Active State: High OUT_SIG: LCINStop SYNC_SIG: StkClear Active state: High TTL-In3 3 6 TTL-Out3 Active State: Low OUT_SIG: StkBusy OUT_SIG: ITSPStrt Active state: High TTL-Out1 4 1 TTL-In1 Active State: High SYNC_SIG: ITSPStrt OUT_SIG: ITSPStop Active state: High TTL-Out2 5 2 TTL-In2 Active State: High SYNC_SIG: ITSPStop OUT_SIG: StkBusy Active state: Low TTL-Out3 6 3 TTL-In3 Active State: High SYNC_SIG: StkClear DB25F GND 11 11 GND DB25F Wash 1 Wash1 + 12 12 Pwr-Out1 + Pwr-Out1 + 12 12 Wash1 + Wash 1 Wash 2 Wash2 + 14 14 Pwr-Out2 + Pwr-Out2 + 14 14 Wash2 + Wash 2 Wash1 - 24 24 Pwr-Out1 - Pwr-Out1 - 24 24 Wash1 - Wash2 - 25 25 Pwr-Out2 - Pwr-Out2 - 25 25 Wash2 - DB25F Wash 3 Wash1 + 12 Wash 4 Wash2 + 14 Wash1 - 24 Wash2 - 25 Top Left Rail: ITSP Bottom Right Rail: LC-Inject
- 11. Proposed Software Modifications • Add the associated PAL objects that support the added PAL accessories. • Add the ITSP PAL objects. • Add the PAL objects that support direct synchronization. • Remove the PAL objects for the removed PAL accessories. • Modify some PAL objects needing customization for ITSP and direct synchronization. • Add CTC PAL Loader to the supervisory computer, if needed. • Add CTC PAL Object Manager (in Service mode) to the supervisory computer, if needed. • Add CTC PAL Cycle Composer to the supervisory computer, if needed. • Load ITSP Opiates Method macros, methods, and sample lists on the supervisory computer. • Setup Cycle Composer to control the ITSP rail. • Setup existing PAL control software to control the LC-Inject rail (if possible).
- 12. Supervisory Control Software Options • Native mode dual rail PAL control software available – Thermo Aria OS (not compatible with ITSP) – Axel Semrau Chronos (not as flexible nor as fast as Cycle Composer) – Gerstel Maestro (not as flexible as Chronos nor as flexible or fast as Cycle Composer) • ITSP compatible PAL control software – CTC Analytics Cycle Composer – Axel Semrau Chronos – Gerstel Maestro – Thermo Xcalibur – Agilent MassHunter – Agilent ChemStation – AB Sciex Analyst – Any other software that can utilize ICC-CE custom cycles We selected Cycle Composer combined with hardwired / hardcoded direct synchronization for this project because of flexibility, speed, and robust behavior.
- 13. ITSP SPE Method • Load 200µL hydrolyzed sample • Wash with 100µL H2O • Wash with 100µL MeOH • Elute with 100µL H2O:ACN:NH2OH(7:3:0.2) • Air Flush • Inject 20µL 4 minute cycle time
- 14. Start Synchronization ITSP Rail LC-Inj Rail SET_OUT_SIG : ITSPStop (Off) SET_OUT_SIG : StkBusy (On) SET_OUT_SIG: ITSPStrt (On) WAIT_SYNC_SIG: LCInStrt SET_OUT_SIG : LCInStop (Off) SET_OUT_SIG : StkBusy (Off) SET_OUT_SIG: LCInStrt (On) WAIT_SYNC_SIG: ITSPStrt
- 15. Cold Stack Synchronization ITSP Rail LC-Inj Rail WAIT_SYNC_SIG : StkClear SET_OUT_SIG : StkBusy (On) Perform ITSP SPE SET_OUT_SIG: StkBusy (Off) SET_OUT_SIG : StkBusy (Off) WAIT_SYNC_SIG : StkClear SET_OUT_SIG: StkBusy (On) Get sample Clean syringe Pre clean syringe and wait at home SET_OUT_SIG: StkBusy (Off) LC Inject and post clean syringe Direct synchronization between the rails allows for overlapping actions, significantly increasing throughput.
- 16. Stop Synchronization ITSP Rail LC-Inj Rail SET_OUT_SIG : ITSPStop (On) WAIT_SYNC_SIG: LCInStop SET_OUT_SIG: ITSPStrt (Off) SET_OUT_SIG : LCInStop (On) WAIT_SYNC_SIG: ITSPStop SET_OUT_SIG: LCInStrt (Off)
- 17. Opiates ITSP SPE Method
- 18. Opiates ITSP LC-Inj Method