you’re doing it wrong: rethinking the application of
TRANSCRIPT
Frederick G. Strathmann, PhD, MBA, DABCC (CC, TC)VP of QA | Assistant Laboratory Director | Director of New Technology & Innovation
NMS Labs, Willow Grove PAemail | LinkedIn | Twitter
You’re Doing it Wrong: Rethinking the Application of Available Mass Spectrometry Technologies for Toxicology
Outline
• Operational Efficiency and How We Measure• High Throughput Mass Spectrometry• Challenges to Conventional Thinking
1. Using an Example of Volume and Menu Growth
2. Marrying a High Throughput System with Mass Spectrometry Capable of “Keeping Up”
The Origin Story
Where Does Mass Spectrometry Fall?
• Continuous Flow
• Centrifugal Analysis
• Discrete Analysis
Theory of Constraints | “The Goal”
• Set in a factory• Main character learns “Socratically” how to manage his
operations from an unlikely source – his old physics professor
• Major epiphany comes form a Boy Scout hike with his son and the infamous “Herby”.
Yale Weis ©123RF
Example Test Process Workflow | 20hrs
Matching Supply with Demand: An Introduction to Operations Management (2013)
Job Shop or Flow Shop?
Matching Supply with Demand: An Introduction to Operations Management (2013)
(c) Robotiq
(c) Robotiq
Flow Shop Mass Spectrometry
Easy to automateEasy to measureEasy to optimize
InflexibleResource intensive (initially)Capacity slow to respond
PRO CON
Easy to startFlexibleCapacity quick to respond
Hard to automateHard to scheduleHard to measure/improve
PRO CON
(c) Robotiq
(c) Robotiq
Increasing Throughput | Example 1
• TDM Assay• Tacrolimus• Sirolimus• Everolimus
• 3 minute inj to inj• 96 well plate• 4 Stds, 4 QC (bookend)• 88 patients max per batch• 5 hours per batch• 2 batches per day or 176 results per day spread
across all three compounds
Adapted from Kocak et al. 2015 PMID: 26526462
Initial Capacity Assessment
Initial Capacity Assessment
Initial Capacity Assessment
A Closer Look at Utilization
Increasing Throughput | Example 1
New Capacity Assessment
Increasing Throughput | Example 2
• Opiate Assay• Morphine, Codeine, 6AM• Hydromorphone, Norhydromorphone• Hydrocodone, Norhydrocodone• Oxycodone, Noroxycodone• Oxymorphone, Noroxymorphone
• 3 minute inj to inj• 96 well plate• 4 Stds, 4 QC (bookend)• 88 patients max per batch• 5 hours per batch• 528 results per day
• 50%, 25%, 10%, 10%, 5%
1
2
3
4
5
Initial Capacity Assessment
Increasing Throughput | Example 2
1
2
3
4
5
New Capacity Assessment
Increasing Throughput | Example 2B
1 2
34
5 Panel 1
Matching Supply with Demand: An Introduction to Operations Management (2013)
New Capacity Assessment B
Increasing Throughput | Panels
Reference Mass: 121.0509922.0098
Agilent 1260 HPLC + 6230 TOF-LC/MSC18 based LC columnsPositive mode: 104 basic drugs
Courtesy of Dr. Kathryn Smith, ARUP Laboratories
Increasing Throughput | Panels
Column: Waters XSelect CSH C18 3x50 mm, 2.5 µmMP-A/Wash 1: 0.1% acetic acid in waterMP-B/Wash 2: 100% acetonitrileFlow rate: 500 µL/minInjection volume: 5 µL Column temperature: 30 °C
Positive Mode
Negative Mode
Courtesy of Dr. Zlatuse Clark, ARUP Laboratories
Downside to Panels
• Multi Calibrators and Multi QCs may tax instrument limitations while patients will not
• Reagent preparation may become problematic• Adding new compounds requires rework of existing
reagents• Statistical QC issues with low volume, unordered tests• Large size may inhibit modifications (too many moving
parts)• Concessions in compounds added together
Job Shop Mass Spectrometry
(c) Robotiq
Increasing Throughput | No LC
Displaces ions Restores ion path
Example Data | LDTD-DMS-MS/MS
XIC of -MRM (4 pairs): 319.300/191.100 Da from Sample 1 (TuneSampleID) of MT20180322105746.wiff (Turbo Spray IonDrive) Max. 5.0e6 cps.
-28 -26 -24 -22 -20 -18 -16 -14 -12 -10 -8 -6 -4 -2 0 2 4 6 8COV, Volts
0.0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
3.5e6
4.0e6
4.5e6
5.0e6
5.5e6
6.0e6
6.5e6
7.0e6
7.5e6
Intensi
ty, cps
-10.32
Displaces ions Restores ion path
Mycophenolic acid
Mycophenolic acid glucuronide
Courtesy of Joseph Homan, NMS Labs
Capacity Assessment
Job Shop Mass Spectrometry
(c) Robotiq
LDTD + Mobility | Easing Throughput
What Else? | Screen with Reflex
Sample Collected
Screen w/ reflex ordered
Positive Negative
StopConfirmation
Report out NegativePositive Negative
Report out Concentration Report out Negative
Non-Specific Detection Methods
Johnson-Davis et al. (2016). Journal of Analytical Toxicology 40(2), 97-107.
Screen Only – Clinical Toxicology
Doyle, K. and F. G. Strathmann (2016). "Cost and Efficacy Assessment of an Alternative Medication Compliance Urine Drug Testing Strategy." Pain Medicine.
Screen Only | Cost as a Driver
Doyle, K. and F. G. Strathmann (2016). "Cost and Efficacy Assessment of an Alternative Medication Compliance Urine Drug Testing Strategy." Pain Medicine.
Screen Only | TAT as a Driver
McMillin, G. A., et al. (2015). "A hybrid approach to urine drug testing using high-resolution mass spectrometry and select immunoassays." Am J Clin Pathol 143(2): 234-240.
Screen with Reflex to Confirmation
• Ensure sample switches did not occur• Ensure interferences were not present
• Higher confidence in the final result• Useful with low pretest probability
• interpreting the results of a diagnostic test*• selecting one or more diagnostic tests*• choosing whether to act*
a) without further testing (treatment threshold)b) while awaiting further testing
• deciding whether it’s worth testing at all (test threshold)• The vast majority of clinical results are singlet analyses
and will likely be acted upon.
*https://www.cebm.net/2014/03/pre-test-probability/
Diagnostic Algorithms | Pretest Probabilities in Use
Pharmacogenetic Testing
Pain Management
Source: ARUP Laboratories; www.aruplab.com. Accessed July 17, 2018
Panel differences from Labs
Increasing Throughput | The Mega Screen
Reference Mass: 121.0509922.0098
Agilent 1260 HPLC + 6230 TOF-LC/MSC18 based LC columnsPositive mode: 104 basic drugs
Courtesy of Dr. Kathryn Smith, ARUP Laboratories
Unwieldy and Slow Targeted Panels
Reference Mass: 121.0509922.0098
Agilent 1260 HPLC + 6230 TOF-LC/MSC18 based LC columnsPositive mode: 104 basic drugs
Courtesy of Dr. Kathryn Smith, ARUP Laboratories
Increasing Intelligence | Panels
Courtesy of Natalie Rasmussen, Agilent Technologies
Targeted Screen by Rapid LC-TOF for enhanced specificity for Directed Confirmation including creatinine quantitation
Sensitive & Specific Rapid Screening
Rasmussen, N. N., et al. (2017). "Lowering the Bar for Mass Spectrometry: A Comparison between Immunoassay and Rapid Time-of-Flight for Presumptive Screening of Drugs in Urine." The Journal of Applied Laboratory Medicine: An AACC Publication.
Flexible Panels | Smaller Chunks
Courtesy of Brandon Nelson, NMS Labs
Rapid Injection | Trap & Elute
© Agilent Technologies
TOF and QTOF | Striking a Balance
Unknown Unknowns | Retrospective Data Analysis
https://www.forensicscienceeducation.org/resources/nps-discovery/
Traditional ApproachTest the sample one time.Compare the data to the database.Is one of the targeted drugs present? Report positive or negative.
What is Retrospective Analysis?Once a new drug is discoveredRetrospectively interrogate the data filesDetect drugs that were not known at the time of the physical testing.
Summary Points
• Efficiency is different depending upon the context• Capacity assessments at subsets of the analytical testing
system allow greater insight into process bottlenecks• Use of technology can allow us to be more flexible
convisum © 123RF
Acknowledgements
• NMS Labs
• Joseph Homan• Brandon Nelson
• AACC• MSSS
Zdenek Sasek © 123RF