the q exactive – a benchtop orbitrap mass spectrometer · 2.04 2.68 2.73 1.94 2.77 0.31 1.80 1.75...

The Q Exactive – A Benchtop Orbitrap Mass Spectrometer for DMPK

Presented to:5th EBF Open Meeting, Barcelona.

Simon Szwandt Ph.D.Market Development Manager– Pharma & CRO6th November, 2012.

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Agenda

• Triple vs HR/AM• Instrumentation and HR/AM• Q Exactive Bioanalytical Data – Small Molecule and Biologics• Hepcidin Quantitation

3

The Exactive Family

• The Exactive Plus• HCD cell optional• Upgradeable to Q Exactive• Mass range to m/z 6000• Faster polarity switching• Rotated flatapole• New scan modes

• The Q Exactive• Resolving power >140,000• Parent ion selection capability• Spectral multiplexing• UHPLC and nanoflow capable

Superior selectivity & specificity due to HR/AM-MS and MS/MS

Why HR/AM in Bioanalysis?

5

Full Scan – The Most InformationC:\Xcalibur\...\Paroxetine-FS-002 8/3/2012 1:09:58 PMStd 1ng/mL - FS

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2.14 2.170.25

2.45

2.40

2.60 2.62

1.97

0.34

1.60

1.401.32

1.150.48 1.040.18 0.62

NL:3.37E6TIC F: + c ESI Q1MS [150.000-400.000] MS Paroxetine-FS-002

C:\Xcalibur\...\Paroxetine-FS-002 8/3/2012 1:09:58 PMStd 1ng/mL - FS

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2.15

2.602.552.400.25

2.352.102.69

1.96

2.751.891.85 2.79

1.791.73 2.830.30 1.49

1.390.33 1.290.18 0.49 1.161.05

NL: 2.08E4m/z= 329.58-330.58 F: + c ESI Q1MS [150.000-400.000] MS Paroxetine-FS-002

Triple Quadrupole Q ExactiveC:\Xcalibur\...\Paroxetine-QE-FS-03 8/13/2012 2:33:00 PM1

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0.24

2.632.622.50

2.642.49

2.652.342.33

2.30

2.162.15

0.340.34 2.061.981.87

1.841.792.751.74 2.811.701.56 2.871.491.351.271.140.850.37 0.51 0.59 0.730.11

NL:8.46E8TIC F: FTMS + p ESI Full ms [150.00-400.00] MS Paroxetine-QE-FS-03

C:\Xcalibur\...\Paroxetine-QE-FS-03 8/13/2012 2:33:00 PM1

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2.02 2.311.93

NL: 3.05E6m/z= 330.14835-330.15165 F: FTMS + p ESI Full ms [150.00-400.00] MS Paroxetine-QE-FS-03

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SIM – The Highest Possible Signal

C:\Xcalibur\...\Paroxetine-SIM-002 8/3/2012 1:04:52 PMStd 1ng/mL - SIM

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RT: 2.15AA: 36614 2.58

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2.251.991.98

1.82

0.30 1.631.491.351.191.000.43

NL: 2.08E4m/z= 329.58-330.58 F: + c ESI Q1MS [330.079-330.081] MS ICIS Paroxetine-SIM-002

Paroxetine-QE-SIM-05_120813142428 8/13/2012 2:24:28 PM1

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RT: 1.96AA: 846873

NL: 5.56E5m/z= 330.14835-330.15165 F: FTMS + p ESI SIM ms [328.65-331.65] MS ICIS Paroxetine-QE-SIM-05_120813142428

Triple Quadrupole Q Exactive

7

Product Ion Scan

C:\Xcalibur\...\Paroxetine-PI-005 8/3/2012 1:35:31 PMStd 1ng/mL - PI

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2.15

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2.582.56

2.392.23 2.612.04

2.68 2.731.94 2.77

1.800.31 1.751.57 2.901.541.14 1.320.990.07 0.46 0.72 0.950.61

NL:4.97E4TIC F: + c ESI Full ms2 330.081 [50.000-400.000] MS Paroxetine-PI-005

Paroxetine-PI-005 #270-281 RT: 2.11-2.19 AV: 12 NL: 3.91E3F: + c ESI Full ms2 330.081 [50.000-400.000]

50 100 150 200 250 300 350 400m/z

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192.05

70.35 330.17

134.81123.07 151.17212.59108.98 178.02

311.73223.1097.15 163.15 237.33 365.9884.38 279.58 301.86 344.7657.12 383.96

Paroxetine-QE-MSMS-NoIS-001 8/13/2012 2:37:20 PM1

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2.51

2.27 2.292.262.24 2.300.24

0.23 2.572.322.232.63

2.21 2.342.642.342.20

2.652.18

1.88 1.961.88 2.16

1.86 2.151.84 2.121.82 2.11

1.801.77

2.701.731.681.550.28 2.761.381.03 1.280.940.340.06 0.69 0.850.60

NL: 3.68E6TIC F: FTMS + p ESI Full ms2 [email protected] [50.00-355.00] MS Paroxetine-QE-MSMS-NoIS-001

Paroxetine-QE-MSMS-NoIS-001 8/13/2012 2:37:20 PM1

Paroxetine-QE-MSMS-NoIS-001 #1317-1332 RT: 1.94-1.97 AV: 16 NL: 1.29E5F: FTMS + p ESI Full ms2 [email protected] [50.00-355.00]

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330.15046

192.11848

70.06580

57.07061 84.96029151.03907 297.0653098.98461

117.07027 279.05448178.10293163.09186

247.12213139.11194

222.99176

335.11606

271.08592 353.12676303.18369

208.11350 320.20065

Triple Quadrupole Q Exactive

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MSMS (SRM Vs. MSMS)

C:\Xcalibur\...\Paroxetine-SRM-001 8/3/2012 12:09:43 PMStd 1ng/mL - SRM

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RT: 2.18AA: 19656AH: 8804

NL: 8.81E3TIC F: + c ESI SRM ms2 330.081 [192.110-192.112] MS Genesis Paroxetine-SRM-001

Triple Quadrupole Q ExactiveParoxetine-QE-MSMS-NoIS-001 8/13/2012 2:37:20 PM1

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RT: 1.96AA: 237725

NL: 1.58E5m/z= 192.11752-192.11944 F: FTMS + p ESI Full ms2 [email protected] [50.00-355.00] MS ICIS Paroxetine-QE-MSMS-NoIS-001

9

Specificity = Resolution + Mass Accuracy

Resolution: 10k, 30k, 50k, 100k

279.12 279.14 279.16 279.18 279.20m/z

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Ethinyl-Estradiol, 279.17434

Butyl-Phthalate, 279.15909 (ubiquitous background ion)

54 ppmapart

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HRAM or MS/MS

This permitted a direct comparison between the LC-HRMS and LC-MS/MS data. The data indicated that the selectivity of LC-HRMS exceeds LC-MS/MS, if high resolution mass spectrometry (HRMS) data is recorded with a resolution of 50,000 full width at half maximum (FWHM) and a corresponding mass window. This conclusion was further supported by experimental data (MS/MS based trace analysis).

This is to demonstrate that Exactive and HRAM with 50K resolution is equivalent or

better than QQQ.

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Why HR/AM Q Exactive Large Molecule

12

Small vs Large Chemical Characteristics

Characteristic Small LargeEndogenous No Often YesSolubility Hydrophobic HydrophilicPurity Homogeneous HeterogeneousStability Chemical/enzymatic Immunologic/enzymaticStability Drug Drug + ReagentsBasis of Measurement Analyte (Direct) Antigen-Ab reaction (Indirect)Reagents Common and

availableUnique, not commercial or available

Calibration Curves Linear Non-LinearExpected/Common interferences

Yes/knowledge Unknown

Technology LC/MS LBA: ELISA, RIA, ECL, Multiplexing

Development Time Weeks Months

B. De Silva, Bristol-Myers Squibb

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Real High Resolution to Increased Selectivity

Dilution curves

Multiplex SIM Mode:

> Separation of interfering signals: improvement of LOQ (200 x)> Increase in cycle time (~ 2 x)

Resolution : 70 000

Resolution : 140 000

18 amol

4.4 fmol

SDLAVPSELALLK Res : 70 000

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682 682.5 683 683.5

682.371

682.397

682.705

682.899

683.039

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682 682.5 683 683.5

682.232

682.369

682.395

682.703

682.898

683.035

683.370

Zoom in

682.397 682.395682.39 682.39682.397 682.395

Res : 140 000

S. Gallien and B. Domon – Luxembourg Clinical Research, Strassen, Luxembourg

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XIC for DIKCSNILLNNSGQIK at m/z 1007.0435(2+)

Not detected using Full MS

Detected and quantified using SIM MS

Sensitivity Gain by Selected Ion Monitoring (SIM)

• Spectrum noise is dependent on the ratio of compound within a certain ion population

• Spectrum noise is much less in SIM mode

• Sensitivity gain 5 – 10 x with SIM mode

• The gain will be higher in more complex matrices

15

Alprazolam, Full Scan ExperimentAlprazolam

Y = 6366.31+514.015*X R^2 = 0.9967 W: 1/X

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10000001500000

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020000400006000080000

100000120000140000160000180000200000

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Zoom in 50 ppt- 100ppt

50 ppt – 10 ppb250 fg oc - 50 pg oc

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Alprazolam SIM ExperimentAlprazolam

Y = -3135.8+552.216*X R^2 = 0.9982 W: 1/X


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0100002000030000400005000060000700008000090000

100000110000120000

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Q Exactive

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High Throughput HR/AM Quantitation

QQ-Orbi

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The Q Exactive: Hardware Innovations

HCD Cell

OrbitrapMassAnalyzer

S-lensIon Source

Enhanced FT

QuadrupoleMass Filter

C-Trap

20

Benchtop Orbitraps vs QToFs

• All high-resolution, accurate mass instruments trade in three “currencies”

• Scan speed (Hz) – also called “dwell time” which is familiar to triple quad users

• Resolution• Sensitivity

Speed

SensitivityResolutio

n

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• ToF based instruments trade speed for sensitivity• Resolution is based on flight path so it is “fixed” – with a

mass dependence• You can scan at 100 Hz… but data compromised• Typical recommended full scan speed – 150-250 ms• This means ToF scan speeds are 4-10 Hz.

Speed


n

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• Orbitrap based instruments trade speed and resolution• Resolution comes from more time spent measuring the

transient• Slow down to get more resolution• Sensitivity is “fixed” since we pre-fill the C-trap for up to

the Orbitrap scan time (it doesn’t “miss” any ions)

Speed


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Speed


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Plecanatide Peptide Analysis – Pyxant Laboratories

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Plecanatide at 35K Resolution

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Plecanatide at 70K Resolution

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Plecanatide at 35K Resolution Chromatograms

Standard-1-01 - m/z= 841.32 + m/z= 841.82 RT: 3.04 - 3.79 NL: 7.97E4F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.42AA: 131404

3.12

3.21

3.453.32 3.69 3.743.27 3.60 3.653.50

Standard-8-01 - m/z= 841.32 + m/z= 841.82 RT: 3.04 - 3.79 NL: 1.89E7F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.41AA: 35229922

3.50 3.543.13 3.58 3.763.653.20 3.713.343.253.06 3.29

QC-0-01 - m/z= 841.32 + m/z= 841.82 RT: 2.76 - 3.51 NL: 7.33E4F: FTMS + p ESI SIM ms [840.50-842.50]

2.8 2.9 3.0 3.1 3.2 3.3 3.4 3.5Time (min)

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RT: 3.13AA: 91151

3.43

3.453.16

3.33 3.393.183.09 3.283.48

2.99 3.363.313.212.822.78

QC-Low-01 - m/z= 841.32 + m/z= 841.82 RT: 3.05 - 3.80 NL: 1.36E5F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.42AA: 250387

3.113.14

3.17 3.23 3.303.20 3.633.523.273.08 3.69

Blank-02 - m/z= 841.32 + m/z= 841.82 RT: 3.10 - 3.85 NL: 1.33E4F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8Time (min)

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RT: 3.47AA: 8199

3.74

3.57 3.77

3.513.32

3.423.35 3.63

3.213.403.12

3.703.17 3.26 3.30

3.60

QC-High-01 - m/z= 841.32 + m/z= 841.82 RT: 3.07 - 3.82 NL: 2.00E7F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8Time (min)

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RT: 3.45AA: 33238161

3.533.15 3.57 3.62 3.65 3.743.19 3.703.27 3.31 3.803.373.233.12

QC 0 QC Low QC High

Blank 1 ng/mL 250 ng/mL

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Plecanatide at 70K Resolution Chromatograms

Standard-1-01 - m/z= 841.32 SM: 3 RT: 2.95 - 3.70 NL: 8.10E4F: FTMS + p ESI SIM ms [840.50-842.50]

3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.33AA: 113704

3.142.99 3.203.02 3.463.08 3.43

3.383.18 3.583.04 3.633.533.483.11 3.26

Standard-8-01 - m/z= 841.32 SM: 3 RT: 2.99 - 3.74 NL: 1.05E7F: FTMS + p ESI SIM ms [840.50-842.50]

3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.36AA: 19937890

3.50 3.56 3.60 3.713.64 3.683.15 3.303.05 3.193.02 3.09 3.26

Blank-02 - m/z= 841.32 SM: 3 RT: 2.94 - 3.67 NL: 3.29E4F: FTMS + p ESI SIM ms [840.50-842.50]

3.0 3.1 3.2 3.3 3.4 3.5 3.6Time (min)

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3.33

3.113.193.143.08 3.603.03 3.553.22 3.412.97 3.37 3.613.01 3.29 3.50

3.513.473.26

QC-0-01 - m/z= 841.32 SM: 3 RT: 2.96 - 3.63 NL: 3.77E4F: FTMS + p ESI SIM ms [840.50-842.50]

3.0 3.1 3.2 3.3 3.4 3.5 3.6Time (min)

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3.37

3.413.17

3.273.553.23 3.33 3.60

3.46

3.10 3.193.15 3.583.43

3.02

QC-Low-01 - m/z= 841.32 SM: 3 RT: 3.00 - 3.75 NL: 1.82E5F: FTMS + p ESI SIM ms [840.50-842.50]

3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.37AA: 294038

3.423.233.11 3.293.03 3.733.44 3.603.20 3.683.253.163.07 3.51 3.63

QC-High-01 - m/z= 841.32 SM: 3 RT: 3.02 - 3.77 NL: 9.93E6F: FTMS + p ESI SIM ms [840.50-842.50]

3.1 3.2 3.3 3.4 3.5 3.6 3.7Time (min)

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RT: 3.39AA: 19027842

3.49 3.54 3.693.62 3.753.203.10 3.313.04 3.243.15

QC High

Blank 1 ng/mL 250 ng/mL

QC 0 QC Low

28

841.0 841.5 842.0 842.5m/z

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841.31789

841.81989

842.31868

841.68281842.47785

842.02495

841.12388

841.31988

841.82125

842.32102

High Resolution Ensures Accurate Quantification Using SIM

Theoretical Mass [M+2H]+= 841.31987

841.5 842.0 842.5m/z

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841.36617

841.81920

842.31919

841.44544

841.94947

841.82125

842.32102

ΔPPM = 1.2

Resolution = 35 K Resolution = 70 K

ΔPPM = 2.35

Actual

Theoretical

29

• Validated Quantitation of Hepcidin

A Validated Method for the Definitive Quantitation of Hepcidin-25 in Human

Serum by LC/MS using high resolution, accurate mass MS

• John E. Buckholz, Gary A. Schultz, Barry R. Jones, Kristen M. Bearup, Kathlyn M. Porter, Danielle J. Strong, Johnson Zhang

• Advion BioServices, Inc., Ithaca, NY

Hepcidin

AJP - GI February 2006, vol. 290 no. 2 G199-G203

•Hepcidin is a 25-amino acid peptide hormone and is the central regulator of iron metabolism making it an interesting biomarker for many applications.

•25 amino acids, folded over in a hairpin shape with 4 disulfide bridges at Cys7-Cys23, Cys10-Cys22, Cys11-Cys19, Cys13-Cys14

Internal Standard

•Mouse hepcidin molecular weight = 2752.02 g/mol

•Mouse Hepcidin Sequence = DTNFPICIFCCKCCNNSQCGICCKT

human hepcidin

MethodsPreparation and Extraction Procedure

• Protein precipitation extraction• Ostro block for phospholipid removal• 96 well format• Reversed phase chromatography• Q-Exactive • Human Hepcidin Sequence =

DTHFPICIFCCGCCHRSKCGMCCKT

Hepcidin Product Ion Mass Spectrum

33

200 400 600 800 1000 1200 1400m/z

0

5

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25

30

35

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45

50

55

60

65

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75

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85

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95

100

Rel

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ance

698.2

644.0

1040.1

763.7 981.1501.2109.8 858.9 1145.0353.8

696 697 698 699 700 701m/z

Q1 Scan[M+4H]4+

Isotope pattern confirmationhuman hepcidin

EIIHEP-BM_PSAE_067 10/13/2010 10:57:26 PM 20 67 EIIHEP PSAE STD8 2 1

559 560m/z

0

10

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40

50

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100

0

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ance

558.8130z=?

559.0131z=?

559.2131z=?

559.4131z=?

559.6130z=?

560.0130z=?

558.8123z=5

559.0122z=5

559.2120z=5

559.4119z=5

559.6117z=5

560.0107z=5

NL:2.62E2

C113 H170 N34 O31S 9 +H: C113 H173 N34 O31S 9

p (gss, s /p:40) Chrg4,1R: 40000 Res.Pwr . @FWHM

NL:2.68E5EIIHEP-BM_PSAE_067#240-249 RT: 3.32-3.41 AV: 10 T: FTMS {1,1} + p ESI Full lock ms [400.00-1800.00]

698 700m/z

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100

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698.2644z=?

698.5145z=?

698.7645z=?

699.0145z=?

699.2645z=?

699.7644z=?

698.2641z=4

698.5137z=4

698.7634z=4

699.0133z=4

699.2635z=4

699.7630z=4

NL:5.23E3

C113 H170 N34 O31S 9 +H: C113 H173 N34 O31S 9



930 931 932 933m/z

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930.6834z=?

931.0169z=?

931.3502z=?

931.6836z=?

932.0169z=?

930.6846z=3

931.0180z=3

931.3512z=3

931.6843z=3

932.0175z=3

NL:2.62E2

C113 H170 N34 O 31S 9 +H: C113 H173 N34 O 31S 9



Theoretical mass spectrum @ 40,000 FWHM resolution parameter

Experimental mass spectrum at 256 ng/mL

[M+5H]5+

[M+5H]5+

[M+4H]4+

[M+4H]4+

[M+3H]3+

[M+3H]3+

34

HRAM hepcidin @ 30,000 FWHMTop 6 isotopes summed using +/- 5 ppm mass tolerance

35

0.0 1.0 2.0 3.0 4.0 5.0

Time (min)

4.09E5

8.05E5

1.67E5

[M+5H]5+

[M+4H]4+

[M+3H]3+

Human Hepcidin extracts from serum

QQQ m/z 698.2 > m/z 1040.0

Q-E SIM – [M+4H]4+ Top 6 Sum

3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4.0Time (min)

0

20

40

60

80

100

Rel

ativ

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bund

ance

0

20

40

60

80

100

Rel

ativ

e A

bund

ance

0

20

40

60

80

100

Rel

ativ

e A

bund

ance

7.28E1

4.21E2

1.20E3

3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4.0Time (min)

0

20

40

60

80

100R

elat

ive

Abu

ndan

ce0

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Rel

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0

20

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60

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Rel

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1.89E5

1.80E5

1.77E5

Blank

Std 12ng/mL

Std 24ng/mL

Blank

Std 12ng/mL

Std 24ng/mL

0123456789

1011

0 200 400 600 800 1000 1200 1400 1600 1800 2000

Inst

rum

ent R

espo

nse

Nominal Conc.

Linear Dynamic Range HRAM-MS

human hepcidin using 140,000 FWHM resolution

0.000.020.040.060.080.100.120.140.160.180.20

0 5 10 15 20 25 30 35

Inst

rum

ent R

espo

nse

Nominal Conc.

Concentration, ng/mL

Concentration, ng/mL

LINEAR - Weighting Factor = 1/X**2 Slope = 0.005439Intercept = -0.003694r-Squared = 0.9927 LLOQ = 2.0 ULOQ = 2048.0 Mean bias = 5.3Standard deviation = 4.9

37

Accuracy and Precision HRAM-MS human hepcidin using 140,000 FWHM acquisition parameter

LLOQQC LQC MQC HQC ULOQQC 50xDilQCTheor. Conc., ng/mL 2 6 1024 1564 2048 50000

Mean 2.2 6.7 1067.5 1589.3 2160.2 52576.7

S.D. 0.2 0.4 38.7 32.9 55.8 1828.9

%CV 9.1 6 3.6 2.1 2.6 3.5

%RE 110 111.7 104.2 101.6 105.5 105.2

Replicates, n 6 6 6 6 6 6

38

• Same instrument for qualitative and quantitative work. • Operated under multiple modes, i.e., full scan, SIM and MS/MS.• MS tuning simpler than triple quadrupoles. • Enhanced selectivity by exact mass rather than fragment ions. • Increased sensitivity by noise-free baseline and no signal loss from

fragmentation.• Increased mass range good for large molecule quantitation. • Q Exactive can be used for quantitative bioanalysis• Sensitivity (S/N) can be improved• Easy to operate and proven to be potentially beneficial for assays

including: – Difficult to fragment– High background– Multiple charges– High mass

40

Industry Trends

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TP36, Van Natta, Clark

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PK drug discovery

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Thermo Posters at ASMS 2012

42

ASMS – Q Exactive Pharma/Biopharma

• Oral: WOC pm - Regulated Bioanalysis using High Resolution LC/MS: Headache or Opportunity?• Quantitative Pharmacokinetic Sample Analysis and Metabolite Identification of Buspirone

using High Resolution Accurate Mass Spectrometry, Kevin Cook1; Rose Herbold1; Keeley Murphy 1; Panos Hatsis 2, 1Thermo Fisher Scientific, San Jose, CA; 2Novartis, Cambridge, MA

• Evaluation of Resolving Power and Extraction Window with Comparison of Profile and Centroid Modes for High-Resolution Mass Spectrometry Phospholipid Quantitation, Mingkun Fu ; Qing Lu, Millennium: the Takeda Oncology Company, Cambridge, MA

• Oral: MOC pm - Integrated Qualitative and Quantitative LC-MS for Small Molecule Analysis• Development of a Higher Throughput Metabolite Screening Assay in Early ADME Profiling

Using Generic HRMS Acquisition and Automated Data Processing, Anthony Paiva1; Cheryl Klakouski1; Tatyana Zvyaga1; Dieter Drexler1; Jonathan Josephs 1; Harold Weller1; Wilson Shou 1; Ismael Zamora2, 1Bristol-Myers Squibb Company, Wallingford, CT; 2Lead Molecular Design, San Cugat del Valles, Spain

43


• Drug discovery PK analysis using a quadrupole high resolution orbitrap mass spectrometer (Q Exactive): an alternative approach to triple quadrupole , Hongying Gao; Andre Negahban, Pfizer, Inc, Groton, CT

• The Opening of Bioanalytical Space Through Benchtop Accurate Mass Spectrometry: The Analysis of Propofol and its Four Metabolites, Beijing Tan1; Yizhong Zhang2; Andre Negahban3; Hongying Gao4; Christopher Holliman5, 1Pfizer, Inc., Groton, CT; 2Pfizer Inc., Groton, CT; 3Pfizer, Groton, CT; 4Pfizer, Inc, Groton, CT; 5Pfizer Inc, Groton, CT

• High Throughput Bioanalysis of Bile Acids and Bile Salts Using UHPLC Coupled with High Resolution Mass Spectrometers (HR-MS), Jie Ding; Eric Lund; Donald Mckenzie; John Lindsay, Covance Laboratories Inc., Madison, WI

• Utility of High-Resolution-Accurate-Mass to Minimize Interference in Simultaneous Determination of Ribavirin and its Phosphate Metabolites in an In Vitro Assay, Cong Wei; James E. Grace Jr.; Tatyana Zvyaga; Dieter M. Drexler, Bristol-Myers Squibb Company, Research&Development, Wallingford, CT

44


• Utilizing a Non-Targeted HR/AM-MS Method to Accelerate QuantitativeThroughput for In-Vitro Metabolic Profiling, Keeley Murphy ; Kevin Cook; Tim Stratton; Patrick Bennett, Thermo Fisher Scientific, San Jose, CA

• Ultra High-throughput MS Methods for the Discovery of Histone Demethylase Inhibitors, John M. Peltier1; Nicole White1; Priti Gaitonde1; Wendy Broom1; Zhao Bin Kang1; Ji-Hu Zhang1; Michael Acker1; Keeley Murphy2; David Farley1; W. Adam Hill1, 1Novartis Institutes for Biomedical Research, Cambridge, MA; 2Thermo-Fisher Scientific, San Jose, US

• Quantitation of Oligonucleotides In Human Plasma Using Q-Exactive Orbitrap High Resolution MS, Weiwei Yuan1; Laixin Wang1; Min Meng1; Jessica Wang2; Kevin Cook 2; Patrick Bennett2, 1Tandem Labs, Salt Lake City, UT; 2Thermo Fisher Scientific, San Jose, CA

• Robust Ultra High Sensitivity Quantitative Analysis using Low Flow LC/MS/MS and Reduced Phospholipid Matrix Effects. Myth or Reality?, David Humphries1; Roger N. Hayes1; Kevin Cook2; Mark Dreyer2; Xiang He2; Subodh Nimkar2; Patrick Bennett2, 1MPI Research, Mattawan, MI; 2Thermo Fisher Scientific, San Jose, CA

45

ASMS – Q Exactive Pharma/Biopharma• Evaluation of Resolution/Scan Speed Limitations for Quantitation of Small

Molecules by UHPLC-HRMS on the Q-Exactive Platform, Allysen Meymaris1; Richard Lelacheur1; Patrick Bennett2; Kevin Cook2; Xin Zhang1, 1Agilux Labs, Worcester, MA; 2Thermo Fisher, San Jose, CA

• Quantification of Plant Expressed Proteins Using High Resolution LC/MS, Jeffrey Gilbert; Trent Oman; Debbie Schwedler; John Lawry; Jesse Balcer; Suresh Babu Annangudi Palani; Yelena A. Adelfinskaya; Brian Wendelburg; Barry Schafer, Dow AgroSciences, Indianapolis, IN

• Quantitative Bioanalysis using the Q Exactive HRMS: Factors in choosing Resolution and Scan Type, Jack Cunniff1; Chris Yang2; Yujin Wang2; Kevin Cook1; Patrick Bennett1, 1Thermo Fisher Scientific, San Jose, CA; 2Gilead Sciences, Foster City, CA

• Improving the Bioanalysis of Endogenous Bile Acids as Biomarkers for Hepatobiliary Toxicity, Troy Voelker1; Kevin Cook2; Min Meng 1; Patrick Bennett2, 1Tandem Labs, Salt Lake City, UT; 2Thermo Fisher Scientific, San Jose, CA

46


• UHPLC-ESI-HRMS quantitation of metabolites without using reference standards-Impact of mobile phase composition on MS response, Sumithra Katragadda; Dil Ramanthan, Kean University, Iselin, NJ

• LC/SRM Has Come of Age to Quantitate Proteins: Amyloid β isoforms measured in Human Cerebrospinal Fluid and Compared to ELISA, Kwasi Mawuenyega; Tom Kasten; Vitaliy Ovod; Kelly Moor; Ling Munsell; Rose Connors; Wendy Sigurdson; Randall Bateman, Washington University School of Medicine, Saint Louis, MO

• Quantitation of Melatonin and N-Acetylserotonin in Human Plasma by Nanoflow LC-MS/MS and Electrospray LC-MS/MS, Melissa D. Carter; M. Wade Calcutt; Beth A. Malow; Kristie Lindsey Rose; David L. Hachey, Vanderbilt University, Nashville, TN

47

Acknowledgements

• Thermo Fisher Scientific• Patrick Bennett• Kevin Cook• Zhiqi Hao• Keeley Murphy• Jessica Wang• TFS Demo Labs

• Advion Bioservices Inc.• John Buckholz• Gary Schultz• Barry Jones• Kristen Bearup• Kathlyn Porter• Danielle Strong• Johnson Zhang

Visit the site at http://www.planetorbitrap.com

the q exactive – a benchtop orbitrap mass spectrometer · 2.04 2.68 2.73 1.94 2.77 0.31 1.80 1.75...

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