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Mass Spectrometry as the Premier Analytical Tool in Drug Discovery and Drug Development. Walter Korfmacher Exploratory Drug Metabolism Merck Research Laboratories Kenilworth, NJ USA. Outline. New Drug Discovery Challenges Mass Spectrometry Basics

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mass spectrometry as the premier analytical tool in drug discovery and drug development

Mass Spectrometry as the Premier Analytical Tool in Drug Discovery and Drug Development

Walter Korfmacher

Exploratory Drug Metabolism

Merck Research Laboratories

Kenilworth, NJ USA

GBMSDG Talk

outline
Outline
  • New Drug Discovery Challenges
  • Mass Spectrometry Basics
  • Selected In vitro Drug Metabolism Applications
  • Selected In vivo Drug Metabolism Applications
  • Metabolite ID Applications
  • MS Imaging Applications
  • Conclusions

GBMSDG Talk

topics not covered
Topics Not Covered
  • Proteomics
  • Biomarker discovery or assay
  • Metabolomics
  • High Throughput Screening

October 14, 2010

GBMSDG Talk

drug discovery from library to market
Drug Discovery:From Library to Market

Lead Optimization

FDA

Approval

Clinical Testing

Early Discovery

Safety Testing

Compound Libraries

Lead Selection and Optimization

Clinical

Approved Drug

Development

Number of compounds

Stages of Discovery and Development

GBMSDG Talk

October 14, 2010

slide5

NEW DRUG DISCOVERY PIPELINE

Chemistry

Biology--HTS for Receptor Activity

In-vitro Stability Screen

DMPK

Lead Optimization

In-vitro Absorption Screen

P450 Enzyme Inhibition Screen

CARRS Oral PK Screen

Rat IV / PO PK

Dog and Monkey IV / PO PK

Metabolite ID

Rising Dose and Multiple Dose Studies and Safety Screens

Drugs into Development

GBMSDG Talk

the challenge
The Challenge
  • How to deal with the multiple compounds at multiple stages in the drug discovery/drug development pipeline.

GBMSDG Talk

the solution
The Solution
  • LC-MS and LC-MS/MS

GBMSDG Talk

why use mass spectrometry
Why use Mass Spectrometry?
  • Specificity! Non-specific techniques, such as UV and fluorescence, are unable to provide proof of the analyte identity
  • Ease of Use! Modern mass spectrometry software interfaces are easy to use
  • Versatility! MS is both a qualitative and quantitative technique

GBMSDG Talk

the challenge1
The Challenge
  • Choosing the right tool for the task :

OR

Wrench?

MS Toolbox

Hammer?

GBMSDG Talk

slide10

Common MS Tools

GBMSDG Talk

what is lc ms
What is LC-MS?

Liquid Chromatography Coupled to a Mass Spectrometer

(In this case the Mass Spectrometer is a Single Quadrupole instrument)

HPLC Column Ion source Mass analyzer Detector

APCI or ESI

GBMSDG Talk

the challenge compound synthesis
The Challenge—Compound Synthesis
  • At a big Pharma site, one might find hundreds of medicinal chemists who might produce 200-1000 new compounds each week. These have to be assayed.

GBMSDG Talk

the solution lc ms
The Solution—LC-MS
  • The LC-MS system based on a single quadrupole MS is a very useful tool for medicinal chemists who want to know if their synthesis is working correctly—did they make the right compound?

Often this is set up as an open access tool. The chemists set up the run and get results within 24 hours.

GBMSDG Talk

the challenge lead optimization in vitro screening
The Challenge—Lead Optimization In Vitro Screening
  • At a big Pharma site, one might get 100-200 new compounds each week that have to be screened in various in vitro assays. These have to be assayed separately for each screen.

GBMSDG Talk

the solution lc ms ms
The Solution—LC-MS/MS
  • The LC-MS/MS system based on a triple quadrupole MS system is the tool of choice for most quantitative discovery bioanalytical applications.

The application of this tool varies with the screen.

GBMSDG Talk

what is lc ms ms aka triple quadrupole technology
What is LC-MS/MS aka Triple Quadrupole Technology?

Liquid Chromatography Coupled to a Tandem Mass Spectrometer (In this case the Mass Spectrometer is a Triple Quadrupole instrument)

Q1 Q2 Q3

HPLC Column Ion source Mass analyzer Detector

GBMSDG Talk

quantitation gold standard ms ms selected reaction monitoring srm
Quantitation Gold Standard:MS/MS – Selected Reaction Monitoring (SRM)*

Select Fragment Select

precursor ion in Q1 precursor ion in Q2 product ion in Q3

e.g. m/z 216 (Collision Cell) e.g. m/z 174

* Often referred to as Multiple Reaction Monitoring (MRM)

GBMSDG Talk

effects of stages of analysis on signal noise and signal to noise
Effects of Stages of Analysis on Signal, Noise, and Signal-to-Noise

Important Concept!!

S/N

Signal

Noise

1

2

3

4

LC

LC-MS

LC-MS/MS

?

GBMSDG Talk

Stages of Analysis

slide19

LC-TIC

LC-MS

GBMSDG Talk

slide20

LC-MS/MS

10 ng/ml

GBMSDG Talk

discovery in vitro screening
Discovery In Vitro Screening
  • P450 Assay--Enzyme Inhibition Screen
  • Caco-2 cells—Absorption Screen
  • Liver Microsomes/Hepatocytes--Metabolic Stability Screen
  • Plasma Protein Binding

Each In Vitro Assay Uses LC-MS/MS for the analytical step (typically a triple quadrupole MS system).

GBMSDG Talk

high throughput cyp inhibition assay example
High Throughput CYP Inhibition Assay Example
  • Generic LC-MS/MS method
    • 1 minute gradient
    • Monitors 3 substrates in a single LC-MS/MS run
  • Template is used for creating sample list
  • Automatic results calculation and import into Activity Base

GBMSDG Talk

slide23

Method:

Method:

P450 source

P450 source

P450 source

human liver microsomes

human liver

human liver

incubation

incubation

incubation

cocktail (

cocktail

3A4, 2D6, 2C9)

substrates

substrates

substrates

testosterone

testosterone

testosterone

Dextromethorphan

Dextromethorphan

Dextromethorphan

Tolbutamine

Tolbutamine

Tolbutamine

3A4

3A4

3A4

2C9

2C9

2C9

2D6

2D6

2D6

b

b

b

products

products

products

6

6

6

-

-

-

hydroxytestosterone

hydroxytestosterone

hydroxytestosterone

dextrophan

dextrophan

dextrophan

4

4

4

-

-

-

hydroxytobutamide

hydroxytobutamide

hydroxytobutamide

detection

detection

detection

LC

LC

LC

-

-

-

MS

MS

MS

-

-

-

MS

MS

MS

In Vitro Evaluation of CYP Inhibition

Purpose:

Evaluate direct and mechanism-based inhibitors for P450 enzymes

(3A4, 2D6, 2C9) to assess potential for drug-drug interactions.

GBMSDG Talk

in vitro evaluation of cyp inhibition
In Vitro Evaluation of CYP Inhibition

incubation/pre-incubation

Stock solution

from CDC

Serial dilution

1. Coincunation

2. Coincubation

  • three concentrations/cpd: 20 mM, 2 mM and 0.2 mM
  • duplicates/each conc.
  • 30 compounds/set

3. Preincubation

4. Preincubation

GBMSDG Talk

lc ms ms for p450 inhibition screen run time is less than 1 minute
LC-MS/MS for p450 Inhibition Screen-Run time is less than 1 minute

Substrate for CYP3A4, 6-hydroxytestosterone

m/z 305  269

Substrate for CYP2D6, dextrophan

m/z 258  157

Internal Standard

m/z 347  121

Substrate for CYP2C9, 4-hydroxytolbutamide

m/z 287  171

GBMSDG Talk

cyp inhibition screen throughput
CYP Inhibition Screen Throughput
  •  Throughput: ~ 150 compounds /week, ~ 7000 samples/week
  •  Analytical cycle-time: 48 hr from delivery to results
  • NOTE: The advantage for this assay is that it is the same regardless of the test compound—no compound method development needed.

GBMSDG Talk

the challenge metabolic stability screening
The Challenge— Metabolic Stability Screening
  • At a big Pharma site, one might get 100-200 new compounds each week that have to be screened for metabolic stability.
  • The challenge is that LC-MS/MS methods have to be developed for each compound.

GBMSDG Talk

the solution lc ms ms software hardware
The Solution—LC-MS/MS + Software + Hardware
  • Use a generic HPLC method. This works for 80-90% of the compounds.
  • Use a vendor-supplied software tool for automated MS/MS method development, e.g.:
      • QuickQuan™ (Thermo-Fisher)
      • QuanOptimise™ (Waters-Micromass)
      • DiscoveryQuant™ (AB-Sciex)
      • OptimizerTM (Agilent)
  • Use robots for automated sample handling

GBMSDG Talk

metabolic stability assay
Metabolic Stability Assay

Layout shows how a robotic liquid handler can be used to perform the incubation and sample preparation steps in a metabolic stability assay.

GBMSDG Talk

metabolic stability assay1
Metabolic Stability Assay

Scheme shows how a well organized system is needed to provide high throughput metabolic stability data.

GBMSDG Talk

in vivo assays
In Vivo Assays
  • Various types of in vivo assays are performed as part of new drug discovery and development
  • The goal may be to understand absorption (A), distribution (D), metabolism (M), or excretion (E) properties of a compound
  • The goal may be to get pharmacokinetic (PK) information on a compound

GBMSDG Talk

adme pk studies
ADME-PK Studies

Brain-- D

Drug Levels—

LC-MS/MS

MS Image—

MALDI-MS/MS

Plasma— A

Drug Levels—

LC-MS/MS

PK Parameters

Dose NCE (Drug) PO/IV

Liver— D

Drug Levels—

LC-MS/MS

Ref: “Using Mass Spectrometry for Drug Metabolism Studies”

W. Korfmacher, ed., CRC Press, 2005.

ASMS 2010

the challenge in vivo pk screening
The Challenge— In Vivo PK Screening
  • At a big Pharma site, one might get 50 - 100 new compounds each week that have to be screened for in vivo PK.
  • The challenge is that LC-MS/MS methods have to be developed for each compound.

GBMSDG Talk

the solution lc ms ms software planning
The Solution—LC-MS/MS + Software + Planning
  • Use a generic HPLC method. This works for 80-90% of the compounds.
  • Use a vendor-supplied software tool for automated MS/MS method development.
  • Use robots for automated sample handling.
  • Develop a standard PK screening assay.

GBMSDG Talk

in vivo pk screening
In Vivo PK Screening

Source: Drug Discovery Today Volume 13, Numbers 7/8 April 2008

Authors: Bo Liu, Jonathan Chang, William P. Gordon, John Isbell, Yingyao Zhou and Tove Tuntland, Department of Pharmacology, Genomics Institute of the Novartis Research Foundation (GNF), San Diego, USA

GBMSDG Talk

pk screening example carrs
PK Screening Example: CARRS
  • Provide basic pharmacokinetic information for all rapid rat compounds.
    • AUC (0-6hr)
    • Concentration vs Time Profile (0-6 hr)

Throughput: Up to 96 compounds per week

GBMSDG Talk

carrs assay
CARRS ASSAY
  • Protein Precipitation Sample Preparation
  • Generic UPLC conditions (1-2 min run time)
  • Triple Quadrupole MS for assay (Two-point standard curve)
  • Automated MS method development (QuanOptimize)

GBMSDG Talk

preclinical pk studies
Preclinical PK Studies
  • Typical study is one compound dosed oral (PO) and IV (Intravenous) in a laboratory animal. The goal is to get PK parameters in various preclinical species.
  • Typically this produces 50-60 plasma samples.
  • Sample preparation is protein precipitation.
  • A multipoint standard curve is prepared for the assay
  • Analysis is by LC-MS/MS on a triple quadrupole MS/MS system. Usually a generic internal standard is used for the assay.

GBMSDG Talk

discovery pk analysis flowchart
Discovery PK Analysis Flowchart

The MS/MS instrument is normally a triple quadrupole system

GBMSDG Talk

slide42

Samples

SRM

Fail

OK

LC-MS/MS test run (PP Sample prep.)

#1: Matrix effect

Revised Chromatography

#2: Interference

:

LLE

Enhanced mass resolution

SPE

#3: standard curve linearity

Non-routine options

Assay

Rapid MS Method Development ion a Discovery Environment

Xu et al. Anal. Chem.--2005

GBMSDG Talk

October 14, 2010

typical discovery pk assay
Typical Discovery PK Assay

Response ratio

1 – 10,000 ng/mL

GBMSDG Talk

discovery metabolite id
Discovery Metabolite ID
  • Generally this has two components:
    • Lead Optimization Phase--would use unlabelled compounds and in vitro samples to look for major routes of metabolism.
    • Pre-Recommendation Phase—Look for problem metabolites plus in vitro comparison of human to animal metabolism.

GBMSDG Talk

discovery metabolite id1
Discovery Metabolite ID
  • Mass Spectrometry serves two purposes:
    • Finding metabolites-MS systems can be used in various ways to find metabolites in multilple biological matrices (e.g., plasma, bile, urine).
    • Structure elucidation—MS systems can be used to to obtain partial or complete structural identification for the metabolites

GBMSDG Talk

triple quad scan functions to find metabolites
Triple Quad Scan Functions:to find metabolites
  • Neutral Loss Scan
    • no prior knowledge of the parent is required—this is used to look for certain classes of metabolites (e.g., glucuronide, sulfate or glutathione conjugates)
  • Precursor Ion Scan
    • only fragmentation pattern of parent is required—may find unexpected metabolites
  • SRM/MRM
    • the fragmentation pattern of parent is used to predict the fragment ions for likely metabolites—some vendors have software tools that make it easy to build a scan set

GBMSDG Talk

slide47

MS Tools

  • Triple Quadrupole MS systems are the premier analytical tool for LC-MS quantitative assays. They are also useful for metabolite ID applications
  • Q-TOF MS systems are best used for metabolite ID applications and for Imaging MS applications
  • QTrap MS systems are excellent tools for quantitative analyses as well as for metabolite ID applications

GBMSDG Talk

qtrap ms publication in vivo pk samples
QTrap MS Publication In Vivo PK Samples
  • Simultaneously quantifying parent drugs and screening for metabolites in plasma pharmacokinetic samples using selected reaction monitoring information-dependent acquisition on a QTrap instrument:

Li et al. (Covance), RCMS, 1943, 2005.

GBMSDG Talk

slide49

Mass Spectrometer

Sample from in vivo

or in vitro studies

10-50%

Injector

HPLC

50-90%

Radiometric Flow detector

100

100

80

80

60

60

40

40

20

20

0

0

0

0

5

5

10

10

15

15

20

20

25

25

30

30

35

35

40

40

45

45

Time (min)

Time (min)

Radiocarbon Chromatogram

LC/MS Chromatogram

Typical Metabolite Profiling Experiments and Instrumentation

Radioactivity helps to locate the metabolites in the samples

October 14, 2010

GBMSDG Talk

product ion scan

Product ion spectrum of a particular compound

m1+

m2+

m2+

m2+

Product Ion Scan

Select Precursor

Ion

Scan Products

Fragmentation

A key technique for obtaining structural information.

GBMSDG Talk

discovery metabolite id2
Discovery Metabolite ID

(A) HPLC Radiochromatogram of 14C-Gemfibrozil at 25 mm Incubated in Human Liver Microsomes Fortified with NADPH and UDPGA;

(B) Reconstructed Ion Chromatogram;

(C) Full Scan Mass Spectrum of M1 [M-H]- .

Xia, Y.Q. et al., Use of a quadrupole linear ion trap mass spectrometer in metabolite identification and bioanalysis, Rapid Commun. Mass Spectrom., 17(11), 1137, 2003.

GBMSDG Talk

MS/MS spectrum of M1

what is mist
What is MIST?
  • Mass Spectrometry
  • Investigators
  • Security
  • Trust
  • MIST will ensure job security for MS metabolite ID experts

GBMSDG Talk

what is mist1
What is MIST?
  • Metabolites
  • In
  • Safety
  • Testing
  • MIST will ensure job security for MS metabolite ID experts

GBMSDG Talk

key mist points
Key MIST Points

1. Human metabolites that can raise a safety concern are those formed at

    • greater than 10 percent
    • of parent drug’s systemic exposure
    • at steady state.

2. Metabolites identified only in human plasma or

Metabolites present at disproportionately higher levels in humans than in any of the animal test species should be considered for safety assessment.

  • Bottom line: Find human metabolites and then be sure they are “covered” in the tox species.

GBMSDG Talk

new ms tool for finding metabolites hrms
New MS Tool for Finding Metabolites: HRMS
  • High Resolution Mass Spectrometry (HRMS) has become the tool of choice for finding metabolites in complex biological matrices.
    • The improved mass resolution can be used to differentiate metabolites from endogenous background
    • Software tools can use HRMS to find metabolites
    • The accurate mass of a detected metabolite can help to confirm its identity by leading to its empirical formula

GBMSDG Talk

two hrms systems
Two HRMS Systems

LTQ-Orbitrap

The Orbitrap MS provides

mass resolution of 10,000-100,000

The TOF MS provides mass resolution of 10,000-50,000

GBMSDG Talk

why high mass resolution tof ms of sidenafil example
Why High Mass Resolution? TOF-MS of Sidenafil Example

MS window: 0.001 Da

MS window: 0.01Da

MS window: 0.1 Da

MS window: 1 Da

Scan: 200 - 800

GBMSDG Talk

slide58

Use of Mass Defect Filter for Post-Acquisition Processing of Accurate Mass (High Resolution) LC-MS Data.

  • “Fish-out” drug-derived peaks from endogenous peaks in a complex biological matrix
  • Key utility in non-radio-labeled drug-administration
  • An alternative to triple quadrupole tools: neutral loss scan (NLS) and precursor ion scan (PIS)

M. Zhu et al., Drug Metab. Dispos. 2006, 34, 1722-1733

GBMSDG Talk

mass defect filter reference
Mass Defect Filter Reference

J. Mass Spectrom., 2009, 44, 999-1016.

GBMSDG Talk

slide61

Mass Defect Filter Example

(B)

TIC

(C)

Processed MS

14C

Source: JMS 2003, 38, 1110-1112

GBMSDG Talk

slide62

Mass Defect Filter Example

(B)

(C)

Mass spectra of metabolite x at retention time 35.5 min

(A) Full scan spectrum of metabolite x form the unprocessed total ion chromatogram . (B) Detail of (A) in mass range 450-550 Da. (C) Full scan spectrum of metabolite x (the molecular ion was at m/z 503.0737 from the MDF processed total ion chromatogram.

GBMSDG Talk

Source: JMS 2003, 38, 1110-1112

metabolite id software tool bgs nora
Metabolite ID Software Tool: BgS-NoRA

Published in RCMS, 23, 1563 (2009).

GBMSDG Talk

metabolite id software tool bgs nora1
Metabolite ID Software Tool: BgS-NoRA

Mouse urine spiked with diclofenac microsomal incubation sample

TIC

TIC after Background Subtraction

TIC after BgS-NoRA

P = parent

M1, M2, M3 = metabolites

GBMSDG Talk

metabolite id software tool bgs nora2
Metabolite ID Software Tool: BgS-NoRA

Mouse urine spiked with diclofenac microsomal incubation sample—peak at 7.8 min

Unprocessed data

Mass spectrum after data processed with BgS-NoRA

GBMSDG Talk

which tool is best for metabolite id
Which tool is Best for Metabolite ID?

Published in RCMS, 24, 939 (2010).

GBMSDG Talk

tk study support
TK Study Support
  • Typical study is one compound dosed oral (PO) in a laboratory animal. The goal is to get TK (toxicokinetic) parameters.
  • This is a GLP (Good Laboratory Practices) study.
  • Sample preparation is typically SPE.
  • A multipoint standard curve is prepared for the assay.
  • Analysis is by LC-MS/MS on a triple quadrupole MS/MS system. Usually a SIL (stable isotope label) internal standard is used for the assay.

GBMSDG Talk

clinical pk study support
Clinical PK Study Support
  • Typical study is one compound dosed oral (PO) in humans. The goal is to get PK parameters.
  • This is a treated as a GLP (Good Laboratory Practices) study.
  • Sample preparation is typically SPE.
  • A multipoint standard curve is prepared for the assay.
  • Analysis is by LC-MS/MS on a triple quadrupole MS/MS system. Usually a SIL (stable isotope label) internal standard is used for the assay.

GBMSDG Talk

impurities and degradants
Impurities and Degradants
  • These studies are performed to support safety studies or clinical studies.
  • The goal is to measure any significant impurities or degradants that are in the pharmaceutical test compound batch used for these studies.
  • Generally, one would use a combination of triple quadrupoles as well as QTOF MS systems as well as the Orbitrap MS system to characterize these compounds.

GBMSDG Talk

slide72

Detector

Quadrupole

Mass Filter

Collision

Cell

Time-of-

Flight

Analyzer

h

+

MS Imaging using the MALDI QqTOF

GBMSDG Talk

slide73

Rat Brain Tissue Slice

--Rat dosed with clozapine

Radioautographic Image

MALDI-MS/MS Image

1000 µm

Optical Image

  • MALDI-MS/MS Image is in good agreement with the radioautographic image

Hsieh Y, et al., Rapid Commun Mass Spectrom. 2006;20(6):965-72.

GBMSDG Talk

slide76

Mouse Whole Body Image—Mouse Dosed with Terfenadine

Optical image of whole-body mouse

slice

  • 100 mpk p.o.
  • and sacrificed
  • 4 h postdosing

Source: Chen et al.,

Drug Metab. Lett., 2,

1-4 (2008)

Ion image of terfenadine (parent).

stomach

GI tract

liver

Ion image of fexofenadine (metabolite).

Result: These MS images allow us to “visualize” first-pass metabolism.

GBMSDG Talk

advion nanomate lesa liquid extraction surface analysis system
Advion Nanomate LESA (Liquid Extraction Surface Analysis)System

Description of Technology

Automated liquid extraction-based surface sampling technique utilizing the robotic Advion Nanomate chip-based nanoelectrospray platform. Method invented at Oakridge National Laboratory (ORNL). Developed and commercialized at Advion.

Liquid microjunction created between the robotically controlled pipette tip dispensing solvent and surface (e.g. tissue section, blood spots on paper).

The microfluidics chip contains an array of nanoelectrospray nozzles etched in a silicon wafer eliminating carryover as one tip and one nozzle is used per sample.

Can be used for a variety of samples including tissue sections, dried blood spots on paper, samples on MALDI plates for complimentary information by electrospray ionization (ESI), TLC plates, and other planar separation media.

Advion Nanomate

ESI Chip

Liquid microjunction between pipette tip and tissue surface

http://www.advion.com/biosystems/triversa-nanomate/LESA/index.php

Schematic of chip based nano-ESI infusion

October 14, 2010

GBMSDG Talk

slide78

Whole Body Distribution of a Drug and itsMetabolites by QWBA vs LESA

7.5 mg/kg IV [3H] Propranolol

T

QWBA (Quantitative Whole Body Autoradiography):QWBA study with metabolite ID by radioprofiling in conjunction with nanospray MS or accurate mass MS at Merck

T

T

Male CD-1 Mice

7.5 mg/kg IV ‘Cold’ Propranolol

MS Tissue Imaging/Sampling:Sections transferred to glass slides with UV-activated adhesive or collected on adhesive tape and sent to ORNL for MS tissue imaging/profiling

Male CD-1 Mice

slide79

QWBA Results Confirmed High Levels of [3H] Propranolol Related Material in Brain, Lung, Liver, and Kidney

40 µm Mouse Whole Body Sagittal Section: 60 min post [3H]Propranolol IV Dose

QWBA

Brain Lung Liver Stomach Kidney20 µM-eq 40 µM-eq 21 µM-eq 31µM-eq 45 µM-eq

0 100

Autoradioluminograph [3H]Propranolol Drug Related Material

October 14, 2010

GBMSDG Talk

identification of 3 h drug related material drm in tissues
Identification of [3H] Drug Related Material (DRM) in Tissues

Unchanged parent detected in lung and brain.

Major metabolites in liver and kidney identified as hydroxypropranolol glucuronide metabolites by LC-MS/MS-rad.

October 14, 2010

GBMSDG Talk

normal operation using the advion nanomate system
NormalOperationusing the Advion Nanomate System

Mass spectrometer

Sampling tip

Nozzle

Aspirate sample

Transfer sample

Apply HV, spray voltage

Sample

October 14, 2010

GBMSDG Talk

operation using the nano m ate system for surface sampling ornl invention advion lesa
Operation using the Nanomate Systemfor Surface Sampling – ORNL Invention (Advion LESA)

Mass spectrometer

Sampling tip

Nozzle

~ 1 mm spot size

Aspirate solventDispense solvent on sampleAspirate sample solutionTransfer sampleSpray sample

Sample

On

Surface

Solvent

Vilmos Kertesz, Gary J. Van Berkel „Fully Automated Liquid Extraction-Based Surface Sampling and Ionization Using a Chip-Based Robotic Nanoelectrospray Platform” J. Mass Spectrom., 2010 Mar;45(3):252-60.

slide83

LESA: MS for Detection of Propranolol (7.5 mg/kg IV) and a Major Metabolite in Tissues

  • Rapid and automated technique to sample tissues including ones on tape used for QWBA.
  • Successful detection of propranolol and its major glucuronide metabolite not seen by DESI-MS.

Lung

Lung

Kidney

Brain

Brain

Liver

Kidney

Stomach

Muscle

Muscle

Liver

Stomach

Dosed tissue

Control tissue

Propranolol

Propranolol

stomach

brain

kidney

lung

muscle

liver

brain

liver

kidney

lung

stomach

muscle

Hydroxypropranolol glucuronide

Hydroxypropranolol glucuronide

t,min

t,min

conclusions
Conclusions
  • Mass spectrometry is used at multiple stages of new drug discovery and development. Various types of mass spectrometers are utilized including single quadrupoles, triple quadrupoles, Q-Traps, Q-TOFs and Orbitrap MS systems. The need for the multiple types of MS system is due to the variety of assays that are mandated at the different stages in the new drug discovery process.

GBMSDG Talk

slide85

MS Reference Books

2009

2008

2009

2005

Available at Amazon.com

acknowledgments thanks to the following for one or more slides
Waters-MicroMass

Thermo-Fisher

AB-Sciex

Agilent

Marissa Vavrek

Rick King

Swapan Chowdhury

Joanna Zgoda-Pols

Michelle Reyzer

Yunsheng Hsieh

Fangbiao Li

Acknowledgments (Thanks to the following for one or more slides)

GBMSDG Talk

acknowledgements
Acknowledgements

GBMSDG Talk

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