enhancing laboratory productivity through automated online ... · ©2011 waters corporation –...

©2011 Waters Corporation – COMPANY CONFIDENTIAL 1

Anders Feldthus

Enhancing Laboratory Productivity Through Automated Online SPE UPLC-MS/MS


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


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


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


OSM Cartridges


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


Full control in MassLynx


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


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


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


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


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


Inlet Editor View

Multiple Operations

Performed In One Step

Solvent

Selection, Flow Rates And Volumes


Method Overview

An Estimate Of The Extraction Time Required


Improved Sample Clean-up: Removal Of Phospholipids


Improved Sample Clean-up: Ion Suppression


Improved Sensitivity: Online SPE vs Protein Crash

Sirolimus 1ng/mL


Advanced Method Development Mode

Used To Develop & Characterise

Methods

Two Cartridges Connected In

Series And Eluted Separately

Breakthrough Recovery &

Tubing Elution


Method Development Table


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


25% increase in sensitivity using Design

Of Experiment

Manual Method Development Vs DOE: Sensitivity

Mycophenolic Acid 100ng/mL


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


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


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


Questions ?

“A UPLC®-enabled online solid phase

extraction system” THANKS FOR YOUR ATTENTION !

enhancing laboratory productivity through automated online ... · ©2011 waters corporation –...

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