©2011 Waters Corporation – COMPANY CONFIDENTIAL 1
Anders Feldthus
Enhancing Laboratory Productivity Through Automated Online SPE UPLC-MS/MS
©2011 Waters Corporation – COMPANY CONFIDENTIAL 2
Outline
Benefits of Online SPE
The Waters MassTrak™ Online SPE Analyzer — The System — Principles of Operation
Method Development For Online SPE — Use of Design of Experiment Techniques
©2011 Waters Corporation – COMPANY CONFIDENTIAL 3
What Are The Benefits Of Online SPE?
Automation — Removing human factor
Improved Sample Efficiency — No sample loss, all of the analyte eluted onto the column
Improved Sample Clean-Up — Sample concentration — Improved sensitivity
Parallel Processing Of Samples — Reduce time-to-first result
©2011 Waters Corporation – COMPANY CONFIDENTIAL 4
What Is The Waters MassTrak Online SPE Analyzer?
The Waters MassTrak Online SPE Analyzer — A device to provide UPLC-enabled online SPE — Part of the ACQUITY UPLC® Online SPE System
o ACQUITY UPLC Sample Manager o ACQUITY UPLC Binary Solvent Manager o ACQUITY UPLC Column Manager – A o ACQUITY TQ Detector
©2011 Waters Corporation – COMPANY CONFIDENTIAL 5
OSM Cartridges
©2011 Waters Corporation – COMPANY CONFIDENTIAL 6
A Closer Look At The MassTrak Online SPE Analyzer
Left Clamp (Elution)
Right Clamp (Loading)
High Pressure Dispenser (HPD) SPE Solvent
Inlets
Valving to allow switching of
modes
Gripper to move cartridges to & form clamps
2 x 96-well plates/trays of cartridges
©2011 Waters Corporation – COMPANY CONFIDENTIAL 7
Full control in MassLynx
©2011 Waters Corporation – COMPANY CONFIDENTIAL 8
Modes Of Operation
UPLC Mode — Online SPE can be bypassed to allow UPLC operation
Sample Extraction Mode (Online SPE Mode) — Processing of samples in matrix — Primary use of system
AMD Mode (Advanced Method Development Mode) — Method characterization (breakthrough, recovery & tubing
elution)
2D SPE Mode
©2011 Waters Corporation – COMPANY CONFIDENTIAL 9
Principle Of Operation: Sample Extraction Mode
Gripper Places Cartridge In Right Clamp — Specified in sample list
HPD Applies Conditioning And Equilibration Solvents To Cartridge
Typically Use 0.1-0.5 mL Volumes At 3-5 mL/min
©2011 Waters Corporation – COMPANY CONFIDENTIAL 10
Principle Of Operation: Sample Load
Sample Manager Draws Sample During Conditioning And Equilibration Steps
Loop Brought In Line With Loading Solvent From HPD
Sample Pushed Through Cartridge, Analyte Trapped And Stream Diverted To Waste
©2011 Waters Corporation – COMPANY CONFIDENTIAL 11
Principle Of Operation: Sample Wash
HPD Supplies Wash Solvents Across The Cartridges
Up To 23 SPE Solvents Can Be Used — Mix ratios of 2 solvents
can be used in steps of 10% for method development
— Applies to all HPD operations
©2011 Waters Corporation – COMPANY CONFIDENTIAL 12
Principle Of Operation: Sample Elution
Gripper Moves Cartridge To Left Clamp For Elution Step
Elution Time Is Defined In The Inlet Editor — Selectivity of extraction
HDP Supplies Clamp
Flush Solvent To Left Clamp Tubing — Cf. weak wash solvent — Numerous steps can be
applied
©2011 Waters Corporation – COMPANY CONFIDENTIAL 13
Inlet Editor View
Multiple Operations
Performed In One Step
Solvent
Selection, Flow Rates And Volumes
©2011 Waters Corporation – COMPANY CONFIDENTIAL 14
Method Overview
An Estimate Of The Extraction Time Required
©2011 Waters Corporation – COMPANY CONFIDENTIAL 15
Improved Sample Clean-up: Removal Of Phospholipids
©2011 Waters Corporation – COMPANY CONFIDENTIAL 16
Improved Sample Clean-up: Ion Suppression
©2011 Waters Corporation – COMPANY CONFIDENTIAL 17
Improved Sensitivity: Online SPE vs Protein Crash
Sirolimus 1ng/mL
©2011 Waters Corporation – COMPANY CONFIDENTIAL 18
Advanced Method Development Mode
Used To Develop & Characterise
Methods
Two Cartridges Connected In
Series And Eluted Separately
Breakthrough Recovery &
Tubing Elution
©2011 Waters Corporation – COMPANY CONFIDENTIAL 19
Method Development Table
©2011 Waters Corporation – COMPANY CONFIDENTIAL 20
Secondary Agent In Renal Transplant Management And As A Primary Agent In The Treatment Of Lupus Patients And IgA Nephropathy
Increasing Demand For TDM Due To Hematological And
Gastrointestinal Toxicity
Comparison of manual versus DOE method development
DOE Optimization Example: Mycophenolic Acid
©2011 Waters Corporation – COMPANY CONFIDENTIAL 21
25% increase in sensitivity using Design
Of Experiment
Manual Method Development Vs DOE: Sensitivity
Mycophenolic Acid 100ng/mL
©2011 Waters Corporation – COMPANY CONFIDENTIAL 22
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Manual Design of Experiment
Volu
me
(mL)
MethanolWater
Manual Method Development Vs DOE: Solvent Use
©2011 Waters Corporation – COMPANY CONFIDENTIAL 23
IVD-Labeled Products Value to Clinical Customers
What are the implications of a medical device (IVD label) designation on Waters products?
• Designates that Waters manufactures medical devices (not general laboratory or research use only devices)
• Signifies that the products have been designed, developed, and manufactured in accordance to and compliance with the Quality System Requirements (US) and In Vitro Diagnostics Directive (EU) ISO 13485 EU IVDD 98/79/EC
• The regulations are intended to provide the market with safe and effective products
• Demonstrates that the vendor (Waters) is committed to ensuring high quality products for clinical laboratories intended to generate high quality patient results
• Waters manufacture to IVD CE certificated Standards.
©2009 Waters Corporation | COMPANY CONFIDENTIAL ©2011 Waters Corporation – COMPANY CONFIDENTIAL
Acquity Online SPE Manager
Official launch April 2012
Latest Advances : on line sample preparation
©2011 Waters Corporation – COMPANY CONFIDENTIAL 25
Conclusions
MassTrak Online SPE Analyzer — Automated online SPE – all of the benefits of SPE and more — Removing the human factor — Parallel processing & improved time-to-first result — Sample Clean-up — Sample Concentration
Simple Method Development Process — Quick and easy to develop new SPE methods
o Method development table facilitates DOE analysis — Optimize methods for both response and robustness
*NOTE: The MassTrak Online SPE Analyzer is under development
©2011 Waters Corporation – COMPANY CONFIDENTIAL 26
Questions ?
“A UPLC®-enabled online solid phase
extraction system” THANKS FOR YOUR ATTENTION !