Page 1
©2011 Waters Corporation 1
Beyond Sensitivity
Improving the Performance, Productivity
and Compliance of the Bioanalytical
Assay Process
EBF, November 17th 2011, Hesperia Towel Hotel
Barcelona
Dr Diego Rodriguez Cabaleiro
Bioanalysis Solution Manager
Waters Marketing Europe
Page 2
©2011 Waters Corporation 2
Ultra high sensitivity for
targeted quantification
Information rich qualitative
supporting data
EBF 2010 Xevo TQ-S
ScanWave
Versatility &
Simplicity of operation
Page 3
©2011 Waters Corporation 3
Xevo TQ Xevo TQ-S
Larger sampling orifice
How did we increase sensitivity?
Problems associated with a larger sampling orifice:
1) A disperse ion cloud
2) Higher levels of matrix contamination
Page 4
©2011 Waters Corporation 4
Ele
ctr
ic F
ield
Diffuse Ion
Cloud
Maximising signal
Maximising robustness
Dealing with a Larger Orifice
Page 5
©2011 Waters Corporation 5
Increase in sensitivity!
Fenuron ESI+ 30 7
Metamitron ESI+ 32 15
Acephate ESI+ 27 7
Chlortoluron ESI+ 27 8
Aldicarb ESI+ 27 6
Demeton S Methyl ESI+ 26 9
Phoxim ESI+ 64 19
Kresoxim Methyl ESI+ 64 4
Azinphos Methyl ESI+ 42 6
Azoxystrobin ESI+ 45 4
Dimethoate ESI+ 23 10
Acetamiprid ESI+ 30 28
Fluticasone ESI+ 30 3
Formoterol ESI+ 39 4
Nefadazone ESI+ 28 3
Desmopressin ESI+ 129 25
Salmeterol ESI+ 41 8
Alprazolam ESI+ 21 13
Reserpine ESI+ 25 5
Ibuprofen ESI- 13 16
Prostaglandin E2 ESI- 30 37
Mean Difference 38 11
Relative
Peak Area
Relative
S:NCompound Name
Ionisation
Mode
Page 6
©2011 Waters Corporation 6
min
2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40
%
0
100
MRM of 5 channels,ES+
504.287 > 293.188
180511_spe_jm_B_V_022 Smooth(SG,2x1)
0.75 0.375
7.162e+0053.68
Fluticasone D3 (1)
3.06
11861.36
623557
2.42 2.772.52
2.82
3.35
min
%
0
100
MRM of 5 channels,ES+
501.28 > 293.185
180511_spe_jm_B_V_022 Smooth(SG,2x1)
0.75 0.375
6.864e+0042.83
2.16
2.07
2.502.432.33
2.25
2.54
2.692.57
Flut 293
3.07
1002.32
44254
2.91
2.95 3.16 3.19
3.283.40
3.49 3.61
4.334.053.94
Compound name: fluticasone
Correlation coefficient: r = 0.999364, r̂ 2 = 0.998729
Calibration curve: 0.663497 * x + 2.14527
Response type: Internal Std ( Ref 2 ), Area * ( IS Conc. / IS Area )
Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None
pg/mL0 100 200 300 400 500 600 700 800 900 1000
Response
0
100
200
300
400
500
600
Xevo TQ-S Fluticasone (0.37 pg/mL in Plasma)
Standard
Internal Standard
4 orders of magnitude linear dynamic range
Page 7
©2011 Waters Corporation 7
Great success and great feedback for more improvements!
Page 8
©2011 Waters Corporation 8
Key Customer Challenges in Regulated Bioanalysis
Regulated Bioanalysis
Quality and Consistency
of results
Evolving analytical demands
Sensitive, Robust, meets
assay reqs
Increasing regulatory demands
->Up time
<-Cost per sample
Page 9
©2011 Waters Corporation 9
Chemistry
ACQUITY Family
Xevo Family
Informatics
Current Individual Offerings
Page 10
©2011 Waters Corporation 10
More Innovation Required
Sensitivity potential of the system not realized
Plenty of room to maximize
Existing platforms in place but with compromise
System Performance
Productivity
Validation & Security
System solution
Page 11
©2011 Waters Corporation 11
2011 Introducing Next Generation Regulated Bioanalysis System Solution
• Best in class
Sample Preparation Solutions
• The fastest and most resolving LC without compromise in robustness
ACQUITY UPLC® I-Class
• Different choices for best performance
ACQUITY UPLC Columns
• Most sensitive MS
® TQ-S
• Compliance ready, Interactive workflow-driven data platform
UNIFI
• Installation, maintenance & training
• Compliance services
Service
Page 12
©2011 Waters Corporation 12
Sample Preparation Solution
•Improves reproducibility & throughput vs LLE and PPT
•Combined PPT and superior phospholipid removal
•Highest sensitivity, selectivity and reproducibility
•Minimal matrix effects
•HLB and mix-mode & all formats available
•Straight forward method development strategy
•Specific biomolecules application kits
•Reduce sample handling vs classic PPT (no centrifugation…)
•Particulate free filtrate
•Automated, simple method development, easy workflow and high troughput (96 well plates)
Page 13
©2011 Waters Corporation 13
Comparison of Phospholipids Remaining after Various Sample Preparation Techniques
MRM of m/z 184-184
Time 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80
%
0
100
0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80
%
0
100
0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80
%
0
100
0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80
%
0
100 184.4 > 184.4 (Lipid 184) 2.00e8
2.88 2.29 2.21
2.10
1.90
2.60 2.78 2.72
184.4 > 184.4 (Lipid 184) 2.00e8
2.80 2.27
1.90
2.62 2.56 2.68
184.4 > 184.4 (Lipid 184) 2.00e8
1.96 1.90 1.77
184.4 > 184.4 (Lipid 184) 2.00e8
2.84 2.21 1.96 1.75 1.42 1.38
1.32
1.63 1.51
PPT
Ostro™
LLE with MTBE
LLE with 5% NH4OH in MTBE
Page 14
©2011 Waters Corporation 14
Has minimized dispersion to enhance MS and UV performance
Lowest carryover complementing MS sensitivity and extending MS linear dynamic range
The system’s low dispersion and higher pressure limits, faster cycle-time allows complex separations can be accelerated without compromising chromatographic fidelity
Seamless method trasfer
Robust, proven technology
ACQUITY UPLC I Class
Page 15
©2011 Waters Corporation 15
Fastest Possible Scenario (1 mm Column) ACQUITY vs ACQUITY IClass
Instrument Method Name: 30sec_10_95p600mL_90C ACQUITY
0.1
58
0.2
05
0.3
51
0.4
19
0.4
57
0.4
83
0.5
03
AU
0.000
0.022
0.044
0.066
0.088
0.110
Instrument Method Name: 20sec_NO hold 10_95_p904mL_90C
0.0
98
0.1
17
0.1
70
0.2
13
0.2
41 0.2
61
0.2
76
AU
0.000
0.022
0.044
0.066
0.088
0.110
Minutes0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70
Average peak capacity at 5σ = 57
Average peak capacity at 5σ = 70
ACQUITY UPLC
ACQUITY UPLC – I CLASS
Ballistic gradient on 1mm x 50mm BEH C18
Due to the 15k psi limit, with the ACQUITY, the flow rate is 0.6 ml/mn
With the ACQUITY I-Class the flow rate is 0.9 ml/mn
Page 16
©2011 Waters Corporation 16
ACQUITY UPLC and Column Choices
Highest resolution and sensitivity (sub 2µm)
Straigh forward method development and transfer
Broad selection of selectivities
Multiple dimensions
Broad range of pH and temperatures
High batch to batch reproducibility
Generic
Polar compounds
Basic compounds
Very polar compounds
Page 17
©2011 Waters Corporation 17
Xevo TQ-S
High Performance and information rich data
Stepwave Large increase in sensitivity (all acquisition modes) Designed to be robust to sample matrix contamination
Scanwave Enhanced product ion spectra
RADAR Targeted MRM at the same time as full scan MS
Versatility Ion Sources
Simplicity of Operation
Engineered Simplicity
Page 18
©2011 Waters Corporation 18
Rapid MS to MRM Switching
Page 19
©2011 Waters Corporation 19
m/z100 200 300 400 500 600 700 800 900 1000
%
0
100
m/z100 200 300 400 500 600 700 800 900 1000
%
0
100253.3
123.0
293.3
397.3
441.4619.6
485.5
329.3
214.1
100.1149.3279.3
251.1
415.3
363.4
546.4
468.6
600.7
Time0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
%
0
100
0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
%
9
0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
%
2
Targeted compound MRM positive ESI Fluticasone
in plasma
Matrix Profile
MS full scan negative ESI
Matrix Profile
MS full scan positive ESI
Qu
alita
tive
Qu
an
tita
tive
Rapid MS to MRM Switching
Page 20
©2011 Waters Corporation 20
Using Dual Scan MRM to Evaluate DBS Card Type To Be Used
DMPK ADMPK A
DMPK BDMPK B
DMPK CDMPK C
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
010210_JoM_DBS_cards_013 2: MS2 ES+ TIC
5.98e91.18
1.030.22
0.32
0.94
1.55
1.421.29
2.411.911.76 2.08 2.24
010210_JoM_DBS_cards_017 2: MS2 ES+ TIC
5.98e9
1.54
0.98
0.22 0.63
1.28
1.40
1.63
2.341.78
1.88 2.052.24
2.41
010210_JoM_DBS_cards_025 2: MS2 ES+ TIC
5.98e91.54
0.23 1.45
1.281.030.90
0.58
2.34
1.73
1.872.07
DMPK ADMPK A
DMPK BDMPK B
DMPK CDMPK C
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
010210_JoM_DBS_cards_013 2: MS2 ES+ TIC
5.98e91.18
1.030.22
0.32
0.94
1.55
1.421.29
2.411.911.76 2.08 2.24
010210_JoM_DBS_cards_017 2: MS2 ES+ TIC
5.98e9
1.54
0.98
0.22 0.63
1.28
1.40
1.63
2.341.78
1.88 2.052.24
2.41
010210_JoM_DBS_cards_025 2: MS2 ES+ TIC
5.98e91.54
0.23 1.45
1.281.030.90
0.58
2.34
1.73
1.872.07
Full scan background
signal
DMPK ADMPK A
DMPK BDMPK B
DMPK CDMPK C
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
020210_JoM_DBS_cards_010 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.10
020210_JoM_DBS_cards_020 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.09
020210_JoM_DBS_cards_033 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.09
DMPK ADMPK A
DMPK BDMPK B
DMPK CDMPK C
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
100
020210_JoM_DBS_cards_010 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.10
020210_JoM_DBS_cards_020 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.09
020210_JoM_DBS_cards_033 1: MRM of 3 Channels ES+ 309.2 > 281 (Alprazolam)
9.61e5
1.09
Analyte Response
Page 21
©2011 Waters Corporation 21
UNIFI 1.5: Scientific Information System for Regulated Bioanalysis
Unites our end-to-end system
solution
One software for Chromatography,
MS & Data Management
Workflow based design adjusts to
roles and capabilities of users
Compliance-ready: Designed for
Annex 11 & Part 11 compliance
Scalable, open architecture
Page 22
©2011 Waters Corporation 22
Waters Service and Support
Project Management
System Design & Configuration
Installation Services
Training
Instrument Qualification
Software Validation
Repair & Maintenance
Waters Quality parts
Method validation
Method translation
Page 23
©2011 Waters Corporation 23
Evolving Analytical Demands - Bioanalysis of Peptides
Regulated Bioanalysis
Quality and Consistency
of results
Evolving analytical demands
Sensitive, Robust, meets
assay reqs
Increasing regulatory demands
->Up time
<-Cost per
sample
Page 24
©2011 Waters Corporation 24
Therapeutic Peptide Method Development Kit
*= data being generated
Maximum recovery = enhanced sensitivity Minimum matrix effects = selectivity and sensitivity
Peptide pI MW % SPE
Recovery % Matrix
Effects
Octreotide 9.3 1019 88 <10%
Angiotensin II 7.35 1046 82 8%
Desmopressin 8.6 1069 104 <11%
Vasopressin 9.1 1084 100 -3%
Goserelin 7.3 1270 100 -2%
Angiotensin I 7.51 1296 109 *
Somatostatin 10.4 1638 94 *
Neurotensin 8.93 1673 114 6%
Bivalirudin 3.87 2180 100 10%
BNP 12 3464 84 *
Teriparatide 9.1 4118 97 9%
Enfuvirtide 4.06 4492 102 *
Page 25
©2011 Waters Corporation 25
MP35N offers a more inert
surface that avoid
adsorptions
Smother internal surface
Updated Tubing Material in I-Class
50x 100x
Conventional SS New MP35N
Page 26
©2011 Waters Corporation 26
m/z 400 600 800 1000 1200 1400 1600 1800
%
0
100 Scan ES+
m/z range of Quadrupoles (2048 amu)
Enfuvirtide
MW 4492
Parent ions
— 3+ at m/z 1498
— 4+ at m/z 1124 Requires at least
1500 amu on first quad
1498
1124
m/z 400 600 800 1000 1200 1400 1600
%
0
100
Bivalirudin
— 2 MRM Transitions
o 1091->650
o 1091->1530 • Requires at least 1500 amu on second quad
Singly charged fragments
650
1091 153
0
Page 27
©2011 Waters Corporation 27
Increasing Regulatory Demands
Regulated Bioanalysis
Quality and Consistency
of results
Evolving analytical demands
Sensitive, Robust, meets
assay reqs
Increasing regulatory demands
->Up time
<-Cost per
sample
Page 28
©2011 Waters Corporation 28
Increasing Regulatory Demands: GLP and 21 CFR 11 Compliance
UNIFI
Embedded relational database
Secure server based workgroup architecture
Comprehensive Audit trail
Electronic signature
Built-in Qualification Center
Central repository of work-flow based SOP’s
Fileless connection to Watson LIMS
Structurally Validated calculation of ISR
Automatic calculation and comparison of Matrix Effect
Factor
WATERS SERVICES
Compliance service and consultancy
Instrument Qualification & Software Validation
Page 29
©2011 Waters Corporation 29
UNIFI Scientific Information System
Page 30
©2011 Waters Corporation 30
Workgroup architecture
<< LABORATORY NETWORK >>
Laboratory Network Appliance
Instrument Control
Server
Presentation Layer
Presentation Layer
Client PC Client PC
Business Layer Database Layer
Instrument Control
Laboratory Network Appliance
Presentation Layer Capture Agent
Client PC
Secure Data Center
Enables Managed Information Sharing & Simplifies Compliance Efforts
Page 31
©2011 Waters Corporation 31
N-tier architecture
DATABASE
APPLICATION LAYER ACQUISITION CONTROL
Reporting Data Processing
Event Center Audit Trails
Document Approval Security
Xevo TQ-S
PORTAL CLIENT
Page 32
©2011 Waters Corporation 32
Reducing Software Validation Barrier
Present Future Desire
Barrie
rs
SO
FTW
AR
E
VA
LID
ATIO
N
GAMP 5 for COTS Leverage Vendor testing Test Summary document Vendor Quality systems Supplier Assessment
Built in Tools for Qualification Qualification Center to manage IQ and OQ of Software and Instruments Portal installation on Clients Built in Tools for
Verification Development of common Validation Scripts Built in tools to automate running and documenting scripts
Validation Consulting Integrate common scripts into a complete validation package customised to Customer protocols
?
Waters Quality System and Professional Services can significantly reduce the Validation effort
Page 33
©2011 Waters Corporation 33
What Will YOU Do With
No compromises. More Choices
Focus on Science
Partnering for the complete solution
Invest for today
and future
Leverage Technology in real time
Page 34
©2011 Waters Corporation 34
Questions
Please visit us at the booth
Product Managers
Senior Application Scientists
Software Specialists
Informatics Solution Consultants
Separation Experts