latest news from agilent liquid phase mass spectrometry solutions · 2017-03-28 · lc/ms portfolio...

Latest News from Agilent Liquid Phase Mass Spectrometry Solutions

Shaun Bilsborough PhD

EMEAI LSAG

Latest News from Agilent Liquid Phase Mass Spectrometry Solutions

Shaun Bilsborough PhD

EMEAI LSAG

LC/MS Portfolio

• 6100 Single Quadrupoles

• 6200 TOF

• 6400 Triple Quadrupoles

• 6500 QTOF

• MassHunter Software

EMEAI LSAG

Key Features

• A mass detector for chromatographers

• Unified LC and MSD design

• Mass detection within existing footprint

• OpenLAB CDS, including QA Compliance &

OpenLAB CDS ChemStation Edition

• Highest robustness, reliability and

performance

The New Mass Selective Detector (MSD)

RoHS Compliant

EMEAI LSAG

New Agilent LC/MSD Series2 models

Best Value Configuration - 6125

• Quick and easy move from UV-

based methods to MS data.

• Benefits of mass detection for the

chromatographer

• Great value for the pharma,

chemistry, start-up and contract

labs for small molecule analysis

• MassHunter Walkup software for non-experts

• ESI source exceeds many detection limits

Most Flexible Configuration - 6135

• Extended mass range (3000) for

Biopharma & life science

applications

• Fastest polarity switching for high-

throughput pharma

• Best choice for trace level detection (w/

optional AJS source)

Available with OpenLAB CDS and OpenLAB Chemstation Edition

G6125B/C(A) G6135B/C(A)

EMEAI LSAG

LC/MSD SpecificationsImproved S/N

*1pg reserpine on-column

**30 ms requires upgrade, without upgrade 300 ms

LC/MSD

Model

ESI+

SIM S/N

RMS*

Jet Stream

ESI

S/N RMS*

APCI+

SIM S/N

RMS*

m/z Max

scan

speed

(u/s)

Polarity

Switching

(ms)

6135 >300:1 >650:1 >100:1 10- 3000 10,400 30

6125 >100:1 NA >100:1 2-2000 10,400 30**

LC/MSD Model Multi-mode

source

APPI Mass accuracy (Da) Dynamic

range

Min SIM

dwell (ms)

6135 Yes Yes +/-0.13 >6x106 5

6125 Yes Yes +/-0.13 >6x106 5

w/ MassWorks

Software

+/- .005 Da Spectral accuracy 98.5

EMEAI LSAG

1260 Infinity II Series LC/MSD Bundle –G6125BW

1260 Infinity II Modules

G6125BA MSD

Software OpenLAB CDS ChemStation Edition

G7111B 1260 Infinity II Quaternary Pump

G7114A 1260 Infinity II Variable Wavelength Detector

G7129A 1260 Infinity II Vialsampler (#63 ICC)

Option #001 Replace quat pump with binary pump

G7112B 1260 Infinity II Binary Pump

Option #002 Replace VWD with DAD

G7117C 1260 Infinity II Diode Array Detector

Option #004 Delete VWD

EMEAI LSAG

LC/MS Portfolio

• 6100 Single Quadrupoles

• 6200 TOF

• 6400 Triple Quadrupoles

• 6500 QTOF

• MassHunter Software

EMEAI LSAG

• Support for all Agilent LCs, GCs, and mass spectrometers

• Superior compound detection for LC/MS with techniques such as tMRM, All Ions MS/MS, and Ion Mobility provides flexibility and confidence

Acquisition

• Key software program for the identification and characterization of compounds in all markets

• Innovative and patented software algorithms extract compounds from data and determine formulas and structure

Qualitative Analysis

• High-throughput quantitation of target compounds

• Easily visualize results using the Batch Table or Compounds-at-a-Glance, then quickly generate reports or upload to a LIMS

Quantitative Analysis

Agilent MassHunter Core Programs

EMEAI LSAG

MassHunter Software for your Key Applications

• Qualitative Analysis for confident identifications and to set up methods

Food

• Quantitative Analysis for fast and accurate results

Environmental

• Acquisition for innovative techniques such as tMRM to confirm compounds

Veterinary Drugs

• Personal Compound Database and Libraries (PCDLs) for rapid identification

Forensic Toxicology

• Spectrum Mill for accurate identification of proteins

Proteomics

• BioConfirm for characterization of intact proteins, peptides, and monoclonal antibodies

Biopharma

• Mass Profiler Professional (MPP) for differential profiling

Metabolomics

• Pathway Architect for bringing together genomics, proteomics, and metabolomics

Integrated Biology

EMEAI LSAG

• Ultra-Trace Analysis

• Agilent iFunnel Technology – Attogram zeptomole sensitivity!

• Ideal for biomarker, steroids, and most demanding bioanalytical/clinical assays

6495Ultra High-

End

• Trace Analysis

• Agilent Jet Stream Technology – Sub femtogram sensitivity!

• Ideal for multi-residue pesticide analysis, clinical assay. Upgradeable to 6495!

6470High-End

• Routine Trace Analysis

• Agilent Jet Stream Technology – Sub femtogram sensitivity!

• Ideal for drugs of abuse, chemical and forensics

6460 Mid-High

End

• Routine Analysis

• Lowest cost of ownership, Robust and Reliable

• Excellent sensitivity and productivity

• Ideal for clinical research & drug analysis

6420Entry-Level

Perf

orm

an

ce

6400 Triple Quad Family

EMEAI LSAG

Agilent 6400 QQQ LC/MS

6495 QQQ

6470 QQQ

6420 QQQ

Relative Price

Rela

tive

Pe

rfo

rma

nc

e

6460 QQQ

Ultra-Trace Analysis• Pesticide, Hormones

• Emerging contaminants

• Biomarker, Steroids

• Ultra-low Assay Requirements

• Complex matrices

Trace Analysis• Pesticide, Mycotoxins

• Vet drugs, Designer drugs

• Drug of abuse

• Vitamin D

• Bioanalysis

Routine Analysis• PPCPs, PFCs

• Opiates

• Pain management

• Chemical & Forensics

EMEAI LSAG

6470 QQQ TechnologiesEnhanced Performance in a Smaller Space

Agilent Jet Stream Technology

• Thermal gradient focusing

• Efficient desolvation

• Creates an ion rich zone

• Up to 10x gains in sensitivity

• An Ion Detector with High Energy Conversion Dynode and Low Noise

• Improved ion detection3

• A Curved and Tapered Hexapole Collision Cell

• Effective ion collection and transmission

2

• Enhanced Q1 Ion Optics

• Improved ion transmission1

EMEAI LSAG

High Sensitivity for Benzodiazepines in Plasma LLOQs and MDLs at Low-pg/mL level on the 6470

• Excellent peak area precision (%RSD) and accuracy are observed at the LLOQ level

-HydroxymidazolamLLOQ = 5 pg/mLS/N = 13.6

MidazolamLLOQ = 5 pg/mLS/N = 35.2

-HydroxyalprazolamLLOQ = 5 pg/mL

S/N = 5.4

AlprazolamLLOQ = 5 pg/mL

S/N = 9.2

CompoundLLOQ

(pg/mL)%Accuracy

%RSD(n = 7)

MDL(pg/mL)

-Hydroxymidazolam 5 100.7 6.9 1.1

Midazolam 5 114.6 5.3 0.84

-Hydroxyalprazolam 5 99.4 9.9 1.6

Alprazolam 5 96.1 7.5 1.2

EMEAI LSAG

Confident Quantitation over 5 Orders of Dynamic Range

• Excellent linearity (R2 > 0.995), accuracy and precision are achieved at all levels including LLOQ

Levels %RSD (n = 7) %Accuracy

5 pg/mL 5.32 114.6

25 pg/mL 2.76 96.6

50 pg/mL 2.53 102.8

250 pg/mL 1.22 98.6

500 pg/mL 0.73 98.7

2.5 ng/mL 0.51 97.6

5 ng/mL 0.34 99.1

25 ng/mL 0.56 96.5

50 ng/mL 0.79 96.4

250 ng/mL 0.75 97.3

500 ng/mL 0.47 101.9

Midazolam, 5 pg/mL – 500 ng/mL

5 Orders of Linear Dynamic Range

R2 = 0.9994

Concentration (ng/ml)0.005 0.01 0.05 0.1 0.5 1 5 10 50 100 500 1000

Rela

tive

Res p

ons e

s 2x10

5E-06

1E-05

5E-05

0.0001

0.0005

0.001

0.005

0.01

0.05

0.1

0.5

Levels %RSD (n = 7) %Accuracy

5 pg/mL 7.54 96.1

25 pg/mL 2.77 96.9

50 pg/mL 2.95 101.9

250 pg/mL 1.62 101.3

500 pg/mL 1.20 101.8

2.5 ng/mL 0.42 99.8

5 ng/mL 1.24 102.3

25 ng/mL 2.15 100.1

50 ng/mL 0.77 99.7

250 ng/mL 2.91 100.0

500 ng/mL 0.62 86.6

Alprazolam, 5 pg/mL – 500 ng/mL

5 Orders of Linear Dynamic Range

R2 = 0.9970

Concentration (ng/ml)0.005 0.01 0.05 0.1 0.5 1 5 10 50 100 500 1000

Rela

tive

Res p

ons e

s 2x10

5E-06

1E-05

5E-05

0.0001

0.0005

0.001

0.005

0.01

0.05

0.1

0.5

EMEAI LSAG

Proven System Robustness in Human Plasma

• Stable peak area response was observed over 1,300 injections of plasma extract and 1 week

of continuous operation

• Average area ratio %RSD = 1.8 was observed with use of internal standard (ISTD)

• Average peak area %RSD = 6.6 was observed for benzodiazepines and metabolites

0

0.2

0.4

0.6

0.8

1

1.2

0 200 400 600 800 1000 1200 1400

Peak A

rea R

ati

o

Injection number

-Hydroxymidazolam

0

0.2

0.4

0.6

0.8

1

1.2

0 200 400 600 800 1000 1200 1400

Peak A

rea R

ati

o

Injection number

Midazolam

0

0.2

0.4

0.6

0.8

1

1.2

0 200 400 600 800 1000 1200 1400

Peak A

rea R

ati

o

Injection number

-Hydroxyalprazolam

0

0.2

0.4

0.6

0.8

1

1.2

0 200 400 600 800 1000 1200 1400P

eak A

rea R

ati

oInjection number

Alprazolam

Area Ratio %RSD=1.3

Peak area %RSD=5.7

Area Ratio %RSD=3.3

Peak area %RSD=7.0

Area Ratio %RSD=1.4

Peak area %RSD=7.2

Area Ratio %RSD=1.3

Peak area %RSD=6.3

EMEAI LSAG

Proven iFunnel Technology: Upgrade 6470!

Agilent Jet Stream Hexabore Capillary Dual Ion Funnel

• Thermal gradient focusing

• Efficient desolvation

• Creates an ion rich zone

• Six capillary inlets

• Samples x10 times more

ion rich gas

• Removes the gas but

captures the ions

• Removes neutral noise

EMEAI LSAG

Scan Speed, dMRM and tMRM…..

Dynamic MRM, Fast MRM Speed and Polarity Switch

• Group MRMs in RT windows

instead of time segments

• 2x shorter cycle time supports

narrow UHPLC peaks

• Supported by very fast MRM

speed (1 ms dwell) and

polarity switch (30 ms)

• tMRM database accelerates

acquisition method setup

• tMRM library – Match Score

for confirmation

• tMRM is Fast and Sensitive

• UHPLC compatible

• Simultaneous Quantitation,

Screening and Confirmation

Triggered MRM, tMRM Database and Library

Match

Score

95.1Trigger

Threshold

tMRM Product

Ion Spectrum

Apex

EMEAI LSAG

6500 QTOF Family

• Agilent iFunnel Technology – Low femtogram sensitivity in complex samples

• Ultra trace analyses for proteomics, metabolomics and glycomics

6550High-End

• Agilent Jet Stream Technology – mid femtogram sensitivity

• Enhanced resolution and mass accuracy for complex samples

• Ideal for impurity analysis, metabolite ID, food safety and environmental screening

6545Mid-Range

• Ease of use and robust operation

• Excellent value for everyday routine use

• Ideal for chemical and forensic applications

6530Entry Level

Maximum sensitivity

Performance with

robustness

Advantages of QTOF

at lower cost

EMEAI LSAG

Ultra-Complex Analysis• Lipidomics

• Proteomics

• Glycomics

• Biophysics studies

• Polymers

• Biopharm structural studies

6500 QTOF Portfolio

6560 IM QTOF

6550 QTOF

6530 QTOF

Relative Price

Rela

tive

“P

erf

orm

an

ce

”

Complex, Trace Analysis• Pesticides, Mycotoxins

• Protein ID/ Biomarkers

• Metabolomics

• Qual/ Quan

• Biopharm characterization

Routine Analysis• PPCPs, PFCs

• Impurity analysis

• Chemical & Forensics

• Quality control

6545 QTOF

Performance based upon

sensitivity and resolving power

EMEAI LSAG

Why Accurate Mass?

m/z

MS

Low Mass Accuracy

m/z

MS

High Mass Accuracy

C12 H14 N9 Cl

C13 H23 N3 P S Cl

C14 H24 N2 S2 Cl

C14 H22 N O3 S Cl

C14 H16 N6 O Cl

C15 H25 O P S Cl

C16 H19 N3 P Cl

C17 H20 N2 S Cl

C18 H21 O P Cl

C13 H23 N3 P S Cl

C14 H24 N2 S2 Cl

C14 H22 N O3 S Cl

C14 H16 N6 O Cl

C15 H25 O P S Cl

C16 H19 N3 P Cl

C17 H20 N2 S Cl

C18 H21 O P Cl

Unit accuracy instrument

e.g. ion trap, QQQ, Q

High mass accuracy

instrument

e.g. QTOF, TOF

EMEAI LSAG

Why Buy an LC-Q-TOF?

To identify compounds with the strongest confidence

Up to 5 degrees of confirmation! Accurate mass, isotopic distribution, product ion spectrum, retention time, IM drift time

Spectrum with theoretical

isotope boxesMirror plot?

Is this safe to eat? Confirm with product ion

spectral matchConfirm with accurate

mass and isotopes

EMEAI LSAG


Analysis of very complex mixtures (cellular digest, biological fluids, environmental)

samples)

Co-eluting compounds with similar masses can be resolved for identification

Picture of complex TIC Extracted complex spectra

Complex TIC Complex Spectra Resolution of

VERY similar MW

ions

EMEAI LSAG


To screen for thousands of targets including unknowns

Screen or profile for thousands of compounds, with high confidence from accurate mass + MS/MS

Full mass range data is collected, so data can be retrospectively queried (doping control, forensics)

EMEAI LSAG

How do we use LC/MS?

United States Secretary of Defense 2001-2006 Donald Rumsfeld

EMEAI LSAG

The Rumsfeld Conundrum

… “as we know, there are known knowns; there

are things we know that we know.

There are known unknowns; that is to say, there

are things that we now know we don't know.

But there are also unknown unknowns – there

are things we do not know we don't know.”

United States Secretary of Defense 2001-2006 Donald Rumsfeld

EMEAI LSAG

Types of Screening and Definitions

“Known Knowns” a.k.a. Target Screening

All Ions

PCDL

Qual/Quant

Customised Reporting.

“Known UnKnowns” a.k.a. Suspect ScreeningAll Ions

PCDL

Find by Formula

Auto or Target MS/MS

PCDL MS/MS Libraries

Molecular Structure Correlator

“UnKnown UnKnowns” a.k.a. Discovery ProfilingMS

Profinder

Mass Profiler Professional

Mass Profiler

Auto or Target MS/MS

PCDL MS/MS Libraries

Molecular Structure Correlator

EMEAI LSAG

Screening Workflow SolutionsScreening Target Suspect Discovery

QQQ Using MRM and tMRM

databases, method can be

quickly developed for

implementation

TOF Using PCDLs and an All-Ions

workflow Quantitation and

Qualification is possible using

four points for confirmation

(Rt, AccMass, Fragments, IP).

Suspect Screening requires

the use of Find by Formula

referring to a suspect PCDL

where MS/MS spectra are

available.

QTOF Using PCDLs and an All-Ions

workflow Quantitation and

Qualification is possible using

four points for confirmation

(Rt, AccMass, Fragments, IP).

Using a Suspect PCDL via All

Ions and or Find-by-Formula

to find proposed compounds.

MS/MS of unqualified

compounds for further

identification via Molecular

Formula Generation or

Structural Correlation.

Samples analysis using Mass

Profiler or Mass Profiler

Professional.

EMEAI LSAG

Databases and Libraries for LC/MSIntegrated with Agilent’s Workflows

Separate & Screen Untargeted w/ All Ions

LC/Q-TOF

Separate & Detect

LC/QQQ

Match spectra

Build Targeted Acquisition Methods

tMRM DatabaseSam

ple

Pre

p,

Colu

mns &

Supplie

sA

pplic

atio

n C

onsultin

g

PCDL

Quantify & Report

MassHunter Quant

Confirm Hits & Report

MassHunter Qual & Quant

EMEAI LSAG

Forensic/ToxicologyLC/QTOF Forensic Toxicology PCDL

LC/QQQ Forensic Toxicology tMRM DB

Environmental

LC/QTOF Water Screening PCDL

LC/QTOF Pesticide PCDL

LC/QQQ Pesticide tMRM DB

Food/Feed Analysis

LC/QTOF Mycotoxins PCDL

LC/QTOF Pesticide PCDL

LC/QTOF Veterinary Drugs PCDL

LC/QTOF Extractables and Leachables PCDL

LC/QQQ Pesticide tMRM DB

LC/QQQ Veterinary Drugs tMRM DB

Portfolio – Databases and Libraries

Life Sciences

LC/QTOF METLIN Personal Metabolite PCD and PCDL

LC/QTOF NIST Library PCDL

LC/QQQ Metabolomics dMRM DB & Method

Pharma LC/QTOF Extractables and Leachables PCDL

EMEAI LSAG

Agilent LC/MS Databases and Libraries

Trust Agilent’s long history in database & library development - Let our mass spectrometry experts work for you!

We did the research - you can save that time!

Use the MassHunter Software suite - streamline your analysis workflow!

Keep your testing current - free upgrades for 3 years!

Rely on Agilent application consulting and our large portfolio of sample prep products and columns – Let our application experts help you!

Instant Productivity

Relevant Content

Help you focus on

what you do best

Ease of Use

Confidence

EMEAI LSAG

Sa 6545XT AdvanceBio LC/QTOF

Introducing the…

6545XT AdvanceBio LC/QTOF

Builds upon 6545A model eXTended mass range and sensitivity

for eXTra large biomolecules

Developed for large

molecules such as intact or

digested proteins

New 6545XT QTOF

EMEAI LSAG

6545XT Development Concept

A single

Q-TOF to

drill down

through

each level

of analysis

for

BioPharma

mAB

Subunit

Peptide

Map

PTM

Intact Protein

EMEAI LSAG

AssayMAP

Bravo

Liquid Chromatography

NEW 6545XT

AdvanceBio QTOF

BioConfirm

Software

Only Agilent has the Complete Solution

Advance

BioColumns

EMEAI LSAG

What is the Advantage of Using Automation?

1. Very high reproducibility / very low variation

2. Robust method progress, no user mistakes

3. Higher quality results with more information

4. More walk-away time – high efficiency

5. Higher throughput

Automation provides major advantages even with low

sample numbers!

EMEAI LSAG

Summary

• New Single Quadrupole MS

• New BioPharma/Large Molecule QTOF

• Application-specific method databases and libraries to confirm identity

of compounds in complex matrices

• Application-specific software to support various applications

• Target analysis of knowns

• Suspect analysis on known unknowns

• Discovery profiling of unknown unknowns

EMEAI LSAG

latest news from agilent liquid phase mass spectrometry solutions · 2017-03-28 · lc/ms portfolio...

Documents