2010 Waters Corporation. Xevo TQ-S for Bioanalysis
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1 Xevo TQ-S for Bioanalysis
2 Key innovations High Performance Stepwave RADAR Scan Wave Versatility Ion Sources Simplicity of Operation Engineered Simplicity
3 The Xevo TQ family Built on the same foundations Xevo TQD Fast data acquisition Routine UPLC-MS/MS RADAR acquisition mode Universal Xevo ion TQD source architecture Routine targeted quantification with information rich qualitative supporting data Xevo TQ MS More sensitive than TQD ScanWave Xevo TQ Enhanced MS product ion scanning Fast data acquisition RADAR acquisition mode Universal ion source architecture High performance targeted quantification with information rich qualitative supporting data Best new product, ASMS 2008 Xevo TQ-S Highest sensitivity Fastest data acquisition ScanWave Enhanced product ion scanning RADAR acquisition mode Universal ion source architecture Ultra high performance targeted Frost quantification and Sullivan with information Product rich Quality qualitative supporting data Leadership Award 2011
4 Xevo TQ-S Ultra high sensitivity for targeted quantification
5 An increase in sensitivity? 100 Xevo TQ-S Enhanced sensitivity Desompressin (Peptide) UPLC/MRM, ESI + Relative ion abundance 129X increase in peak area 25X increase in signal:noise Xevo TQ 0 Time Time
6 An increase in sensitivity? Compound Name Ionisation Mode Relative Peak Area Relative S:N Fenuron ESI Metamitron ESI Acephate ESI Chlortoluron ESI Aldicarb ESI Demeton S Methyl ESI Phoxim ESI Kresoxim Methyl ESI Azinphos Methyl ESI Azoxystrobin ESI Dimethoate ESI Acetamiprid ESI Fluticasone ESI Formoterol ESI Nefadazone ESI Desmopressin ESI Salmeterol ESI Alprazolam ESI Reserpine ESI Ibuprofen ESI Prostaglandin E2 ESI Mean Difference 38 11
7 How did we increase sensitivity? Xevo TQ-S Larger sampling orifice Problems associated with a larger sampling orifice: 1)A disperse ion cloud 2) Higher levels of matrix contamination
8 Ion Block Design Sampling cone aperture Increased to 0.8mm diameter o Approx 5x increase in gas/ion flow New ion block design No supplemental pumping All ions (and gas) enter StepWave guide o Approx 200x increase in gas flow o Up to 200x increase in ion flux
9 Designed to deal with problems associated with a larger sampling orifice Electric Field Diffuse Ion Cloud Problems associated with a larger sampling orifice: 1)A disperse ion cloud 2) Higher levels of matrix contamination
10 Designed to deal with problems associated with a larger sampling orifice Electric Field Diffuse Ion Cloud Maximising signal Maximising robustness
11 SIMION Model No Electric Field 1 mbar N 2 N 2 Flow
12 SIMION Model With Electric Field (25 V between guides) 1 mbar N 2 T-Wave E T-Wave N 2 Flow
13 Ultra High Sensitivity? UPLC/MRM of Verapamil (solvent standard) using an ACQUITY prepared for trace analysis replicates RSD< Zeptomoles fg fg 0.83 of Verapamil Solvent 0 Time Blank 0 Time Time
14 Linearity of Response? Compound name: OH Prog Correlation coefficient: r = , r^2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None APCI+, Hydroxprogesterone Triplicate injections of Standards 50fg to 5,000pg on column Response orders of magnitude Correlation coefficient>0.995 Deviation<15 0 Conc
15 Assay Robustness Verapamil, 10pg/µL spiked into supernatant from 2:1 ACN:Plasma protein precipitation. ACQUITY BEH 2.1x 50mm 1000 on column injections RSD of peak areas < 5
16 Assay Robustness 10ul loop injection 0.5pg/ul Verapamil in human plasma/acn_2 TC8_100721_ MRM of 1 Channel ES+ 455 > 165 (Verapamil) 2.66e7 Plasma injection 2000 Plasma injection 1000 Plasma injection 1 Verapamil, 0.5pg/µL spiked into supernatant from 2:1 ACN:Plasma protein precipitation. 10µL injections ACQUITY BDH 2.1x 50mm >2000 on column injections RSD of peak areas < Time
17 Xevo TQ-S Information rich qualitative supporting data
18 Rapid MS to MRM Switching
19 Rapid MS to MRM Switching Matrix Profile MS full scan positive ESI Qualitative Matrix Profile MS full scan negative ESI m/z m/z Quantitative Targeted compound MRM positive ESI Fluticasone in plasma Time
20 Method Development: Comparison of PPT to SPE Extract Protein Precipitation Full Scan (Extracted Interference) Protein precipitation Background ms scan 2.44e8 SPE Background ms scan 2.61e7 MRM Protein precipitation Monitor Alprazolam MRM 309-> e6 SPE Monitor Alprazolam MRM 309-> e6
21 Method Development: Choosing between two gradients Gradient method MeOH in 0.70 minutes Gradient method MeOH in 2.0 minutes Fluticasone 501> Fluticasone 501> Radar Solvent Blank Radar Solvent Blank Radar Plasma Blank Radar Plasma Blank Phospholipids 184> Time Phospholipids 184> Time
22 Simultaneous MRM and Full Scan Detection of Metabolites MRM of Drug Metabolites Full Scan of Metabolites
23 RADAR & StepWave RADAR Monitor matrix complexity RADAR enables understanding of matrix complexity Intelligent decision making when assessing matrix effects Intelligent decision making when developing methods StepWave Extra sensitivity in MRM mode Reduce matrix effects by simple dilution of complex extracts Maintain ability to achieve LOQs
24 Xevo TQ-S Information rich qualitative supporting data ScanWave
25 ScanWave Technology Potential Energy Low m/z Ion DC Barrier RF Barrier Accumulation Intermediate m/z Ion High m/z Ion Storage Region ScanWave Region Transfer Travelling Wave High m/z Ejection Travelling Wave Medium m/z Ejection Travelling Wave
26 Standard Product Ion Scanning Continuous output of ions by cell Most ions filtered out by MS2 Very inefficient
27 ScanWave Enhanced Product Ion Scanning Ion accumulation Mass selective ion ejection Synchronised with scanning of MS2
28 StepWave Combined with ScanWave ScanWave enhanced Product ion spectrum 100 UPLC/MS/MS 50fg Reserpine on column S/N:RMS=60.14 ScanWave enhanced Product ion scanning chromatogram (m/z 195) m/z Time A combination providing leading-edge spectral MS/MS sensitivity
29 Rapid Mode Switching Precursor Ion Confirmation Scan PIC SCAN: MRM data is used as a specific trigger for the acquisition of a ScanWave enhanced product ion spectrum. Eliminates the need for 2 analytical runs for confirmation MRM Acquisition PIC Scan Acquisition
30 ScanWave Technology MRM ScanWave DS A product ion confirmation (PIC) scan is a useful tool for method development, or when there may be two peaks within a single MRM transition MRM
31 An Alternate Approach to Full Scan Profiling of Metabolites Screen for selected panel of metabolites Commonly reported metabolites User definable Predicts metabolite MRMs based upon Parent Compound MSMS Generates MANY MRMs 1 msec dwell With RADAR, full scan MS may be added to the experiment
32 ( ) P F3 F2 F1 P ( ) + M F3 F2+M F1 imrms : P+M > F1 P+M > F1+M P+M > F2 P+M > F2+M P+M > F3 P+M > F3+M
33 Develop Theoretical MRMs to Investigate MSMS of Parent Compound Identify Product Ions From Parent Compound Select Metabolites from list of probable transformations MS Source and Analyzer Conditions Automatic Product Ion Selection Spectra of Parent Automatically Obtained from MassLynx Calculate MRMs and Write MS Method File imrm Method List of Potential Metabolites from Text File
34 imrm method
35 Simple Metabolite Detection and Confirmation 18_4_ _4 _ ( ) 1: Product Ions of 416 ES > (AZ1_Dealkylation) 5.5 6e m/ z : MRM of 1 Channel ES+ 18_4_ > (AZ1_Dealkylation) 5.66e6100 PIC Spectrum 18_ 4_ (1.871) 2 : Product I ons of 428 ES > (AZ1_Demethylation) e PIC Spectrum m /z : MRM of 1 Channel ES > (AZ1_Demethylation) 3.58e6 [Benefits] Metabolites detected with maximum sensitivity 0 18_4_ Dealkylation _4 _ ( ) 3: Product Ions of 442 ES > ( AZ1_ Parent) 8.9 8e PIC Spectrum Time 3: MRM of 1 Channel ES+ 18_4_ > (AZ1_Parent) 1.08e8 100 Dosed Compound 0 Demethylation Time 5: MRM of 1 Channel ES > (AZ1_Hydroxylation) 3.77e5 18 _4 _ ( ) 5: Product Ions of 458 ES > (AZ1_Hydroxylation) 1.9 2e PIC Spectrum Spectra acquired in the time scale of UPLC peak Data acquired in one analytical run, no need for confirmatory experiment m/ z MRM 442.2> m/ z Hydroxylated Metabolite MRM 458.2> Waters 0.50Corporation Time Time
36 ScanWave & StepWave ScanWave Enhanced product ion spectra Confirm target compound quantitative data with PICS Indicate presence of related metabolites StepWave Enhances signals in all ionisation modes Highest quality of spectra MS/MS data for structural confirmation
37 Xevo TQ-S Versatility & Simplicity of operation
38 Simplicity of Operation IntelliStart Automated MS Resolution & Calibration Checks IntelliStart Automated MRM Method Developer Quanpedia Automated MRM Time Window & Data Rate Scheduling IntelliStart Automated LC/MS System Check Automatically ensure the system is ready to run IntelliStart Automated System Monitoring.
39 Calculating a Spiked Simple Method Matrix Factor XIC Analyte XIC Internal Standard Solvent + Analyte and Internal Standard Extracted Matrix + Analyte and Internal Standard A= 100 A= 90 A= 95 A= 85 MF = A matrix /A solvent MF = 90/100 =.90 MF = A matrix /A solvent MF = 85/95 =.89 IS Normalized MF = MF AN /MF IS IS Normalized MF =.90/.89 = 1.01
40 Alternate Approach for Calculating Matrix Factors LC Column MRM Signal 1) Determine AnalyteProfile AN and IS injected through the column 2) Inject Solvent Blank 3) Inject Extracted Matrix Blank Solvent Blank injected with post column infusion of AN and IS Extracted Blank injected with post column infusion of AN and IS AN and IS infused post-column Analyte = AN Internal Standard = IS
41 Automating Matrix Factor Calculations: Integrated Intellistart Fluidics
42 Matrix Factor Calculation Results (MassLynx 4.1 SCN 737 Time 1 1 Time
43 Source versatility Lorem Ipsum Dolor Sit Amet
44 XEVO Family Ion Sources A common ion source platform Designed to be compatible with HPLC, UPLC and GC applications Designed for performance, usability & serviceability.
45 Source versatility TRIZAIC Source with nanotile APGC ASAP DESI DART
46 Simplicity of Operation & Versatility Universal source architecture Enables the widest range of ionization techniques to be utilized ESI, APCI, ESCi Nano-flow, TRIZAIC APPI, ASAP, APGC Compatible with third party sources (DESI, DART, LDTD, etc) Simple and tool-free Allowing you to change between ion source options in minutes.
47 Xevo TQ-S Summary
48 Xevo TQ-S Key MS Technologies StepWave Large increase in sensitivity (all acquisition modes) Designed to be robust to sample matrix contamination RADAR Targeted MRM at the same time as full scan MS ScanWave Enhanced product ion spectra
49 Xevo TQ-S Designed to Meet Customer Needs Ultra high sensitivity to make: Detection of banned substances easier Monitoring of low exposure drugs easier Blood spot analysis easier It possible to reduce matrix effects by simple sample dilution It possible to work with small sample volumes Information rich supporting data
50 Thank You Xevo TQ-S Lorem Ipsum Dolor Sit Amet Consider Quantitative Assays Previously Beyond Your Reach
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