LC-MS/MS Fundamentals

SCIEX Triple Quad LC-MS/MS Systems for

quantitative workflows
Tan Shin Jowl, PhD | Field Application and Market Development Specialist, SEA
© 2019 AB Sciex
22 Feb 2022
SCIEX LC-MS/MS systems

• SCIEX Triple Quad™ Systems (QQQ)

• Quadrupole time-of-flight (QTOF) Systems
‒ SCIEX Triple Quad 3500 LC-MS/MS System
‒ SCIEX Triple Quad 4500 LC-MS/MS System ‒ SCIEX X500R QTOF System

‒ SCIEX Triple Quad 5500+ LC-MS/MS System ‒ SCIEX X500B QTOF System
‒ SCIEX Triple Quad 6500+ LC-MS/MS System ‒ TripleTOF® 5600+ LC-MS/MS System
‒ SCIEX Triple Quad 7500 LC-MS/MS System ‒ TripleTOF 6600+ LC-MS/MS System
‒ ZenoTOF® 7600 QTOF System
• SCIEX QTRAP® Systems (linear ion trap)
‒ SCIEX QTRAP 4500 LC-MS/MS System
‒ SCIEX QTRAP 5500+ LC-MS/MS System
‒ SCIEX QTRAP 6500+ LC-MS/MS System
‒ SCIEX QTRAP 7500 LC-MS/MS System

2 © 2020 DH Tech. Dev. Pte. Ltd.

INTRODUCING the workhorse in high flow LC

• ExionLC™ 2.0 Systems - enhanced, flexible and

scalable HPLC/UHPLC systems, designed for
analytical flow LC applications

• ExionLC 2.0 System

‒ 12,500 psi HPLC/UHPLC system
‒ For users that don’t need 18,000 psi but still want
UHPLC capability
• ExionLC 2.0+ System
‒ 18,000 psi UHPLC system

3 © 2020 DH Tech. Dev. Pte. Ltd.

How to choose between Triple Quad, QTRAP® or QTOF system?
Radial Trapping

Axial Axial
Trapping Trapping

Radial Trapping Exit Lens

with Grid

Triple Quad™ QTRAP® QTOF X500R

− Quantitation
− Quantitation − Quantitation
− ID with MRM ratio
− ID with MRM ratio − ID with accurate mass
− ID with MS/MS
library searching − ID with MS/MS library
searching
− ID true unknowns
− Retrospective data
processing

Increased confidence in compound ID. © 2020 DH Tech. Dev. Pte. Ltd.

LC/MS Fundamentals

Presentation Objectives
• Why LC-MS
• Sample Introduction with Liquid Chromatography
• Ionization Basics
• How to Calculate Mass
• Isotopic Abundance
• Mass Spectra
• What is a TIC, XIC and BPC
• Quadrupole theory
• LC-MS workflow

© 2019 AB Sciex
Why LC-MS?

What is in My Sample and How Much?

LCMS Detection of B Vitamins

XIC of +MRM (39 pairs): 220.106/202.100 amu Expected RT: 4.6 ID: VITAMIN B5 1 from Sample 3 (DOOG FOOD) of WATER SOL.wi... Max. 8.1e5 cps.

1.29e6
1.25e6
1.20e6
1.15e6
1.10e6
1.05e6
1.00e6
9.50e5
9.00e5
8.50e5
4.38
8.00e5
7.50e5

Intensity, cps
7.00e5
6.50e5
6.00e5
5.50e5
5.00e5
4.50e5
4.00e5
3.50e5
3.00e5
2.50e5
2.00e5
1.50e5
1.00e5
5.00e4
0.00
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0
Time, min

LCMS Quantitation Results

Vitamin DV (mg)
Thiamine Vitamin B1 1.5
Riboflavin Vitamin B2 1.7
Niacin Vitamin B3 20
Pyridoxal HCl Vitamin B6 2
Folic acid Vitamin B9 0.4
Cyanocobalamin Vitamin B12 0.006
Biotin Vitamin B7 0.3
Pantothenic acid Vitamin B5 10

© 2019 AB Sciex
Why LC-MS?

Some Applications Examples

• Food Testing
‒ Pesticides, Nitrofuranes, Mycotoxins, other contaminants
• Environmental
‒ Pesticides, Perchlorate, HAAs, PPCPs, other contaminants
• Forensics
‒ Opiates, Benzodiazepines, THC, other drugs of abuse
• Pharma Applications (Discovery, Development, Production)
‒ DMPK, ADME, Microdosing, Bioavailability

© 2019 AB Sciex
LC-MS Fundamentals

What is a mass spectrometer?

• By measuring the mass of the detected analytes, mass spectrometers
provide enhanced selectivity over other types of detectors.

A mass spectrometer is
essentially a “molecular scale”

• Technically, a mass spectrometer measures the ratio of mass-to-charge

(m/z) of an ionized molecule
‒ For most ‘small molecules’, the charge (z) equals 1
‒ Therefore, for most ‘small molecules’ m/z = mass

© 2019 AB Sciex
LC-MS Fundamentals

Basic LC-MS process

intensity
+
+ + + +
+ + + + + +
m/z

Ionized Ions are sorted The signal is The results are

compounds are by m/z detected and displayed
introduced counted

© 2019 AB Sciex
LC-MS Fundamentals

The LC-MS workflow

Liquid sample is
introduced to ion
source.

© 2019 AB Sciex
Liquid Chromatography

© 2019 AB Sciex
LC-MS Fundamentals

Introduction to liquid chromatography (LC)

• Technique for separation of chemical components of a sample
• The sample is carried by the mobile phase (a flowing solvent) through the
stationary phase, resulting in separation of the components of the sample
• Components elute at different times
• Elution time is based on differential affinity for the stationary phase

Stationary
Mobile phase
phase
Separate components in mixtures

Interaction
Injection Elution

© 2019 AB Sciex
LC-MS Fundamentals

The column
• Main parts: body, end fittings, frits, stationary phase
• Columns dimensions are based on length, internal diameter and particle
size

250 x 4.6 mm

150 x 2.1 mm

30 x 4.6 mm

© 2019 AB Sciex
LC-MS Fundamentals

Simplified Column Selection Guide

© 2019 AB Sciex
LC-MS Fundamentals

The mobile phases

• The mobile phase is
responsible for eluting
compounds from the stationary
phase
• The mobile phase serves two
purposes.
‒ Displace compounds from the

%B
stationary phase causing
elution.
‒ Transport unbound materials
through the column and the
rest of the LC system
Gradient

© 2019 AB Sciex
LC-MS Fundamentals

The mobile phases

Mobile phase A “aqueous”
Mobile phase B “organic”

90% A 90% B 90% A

Compounds elute as % Hold at high % organic to Column

organic increases “clean off” the column Equilibration

© 2019 AB Sciex
LC-MS Fundamentals

Mobile Phase pH and pH Buffers

• pH affects ionization
‒ Silica surface of column Buffers pH Range
‒ Sample components of interest
Formic Acid 2.8 – 4.8

• Buffers Ammonium
2.8 – 4.8
‒ Resist changes in pH and maintain Formate
retention
‒ Improve peak shape for ionizable Acetic Acid 3.8 – 5.8
compounds Ammonium
3.8 - 5.8
Acetate
• Affects Column Life Ammonium
‒ Low pH strips bonded phase 6.6 - 8.6
Bicarbonate
‒ High pH dissolves silica

© 2019 AB Sciex
LC-MS Fundamentals

Why bother with LC if you have an MS/MS?

‒ LC separation provides enhanced specificity, based on retention times

‒ LC separation reduces the number of molecules entering the MS ionization

source at a given time
‒ Reduces the competition for charge (a.k.a. “ion suppression”)

© 2019 AB Sciex
Mass Spectrometry:
Measuring Mass

© 2019 AB Sciex
Measuring Mass - Isotopes

Most Elements Have More than One Isotope

Natural
Element Mass
Abundance
1.0078 99.985%
H
2.0141 0.015
12.0000 98.89
C
13.0034 1.11
14.0031 99.64
N
15.0001 0.36
15.9949 99.76
O 16.9991 0.04
17.9992 0.20
F 18.9984 100
31.9721 95.00
32.9715 0.76
S
33.9679 4.22
35.9671 0.02

© 2019 AB Sciex
LC-MS Workflow

Liquid sample is
introduced to ion
source.

Molecules of interest
get converted to
ions

© 2019 AB Sciex
LC-MS Workflow

Liquid sample is
introduced to ion
source.

Molecules of interest Under an electrical field and

get converted to pressure gradient, ions move
ions through the interface

© 2019 AB Sciex
LC-MS Fundamentals

Measuring Mass-to-charge
• Mass Spectrometers control, sort and detect ions, not uncharged
molecules
‒ Uncharged molecules are called neutrals

• Ions can be positively or negatively charged molecules

‒ Positively charged: gain one or more protons;
‒ Negatively charged: lose one or more protons.
‒ Since ions add or lose a H they will have a different mass than the neutral or
molecule weight of the molecule.

• The Mass Spectrometer displays data two ways:

‒ Spectrum
‒ Ion Chromatogram (x-axis is TIME)

© 2019 AB Sciex
LC-MS Fundamentals

What are ions?

• An ion is a charged form of a • Ions exist in positive and
compound negative forms
‒ Adding or subtracting a proton. ‒ Depending on the compound’s
‒ The proton comes from polarity and chemical structure.
Hydrogen.

H+ attraction OH-
attraction
e- +
e-

Neutral Molecule: M

© 2019 AB Sciex
 MS: Sample Ionisation

How do ions travel?

Ions travel down the ion path due to a pressure and DC voltage gradient.
Q3

Positive
QJet® II
Ion Guide
Q2
LINAC®
Ions
Collision Cell

Orifice Q0 Q1

Orifice (DP)
= 30 V Q0 =
R01 =
-10 V R02 =
-11 V
-30 V RO3 = -33 V
© 2019 AB Sciex
Measuring Mass - Mass-to-Charge

Ions and Calculating Mass

• To calculate the mass of an ion use the following mass-to-charge ratio
formulas.
‒ [M+H]+ /z for Positive ions
‒ [M-H]- /z for Negative Ions
Where
M = molecular mass
z = number of charges or charge state
H± = number of protons involved
i.e. A singly charged positive ion has a mass of [M+H]+

Example: Haloperidol
M = 375; z = 1
Therefore the mass of the positive ([M+H]+)
haloperidol ion is:
(375+1)/1 = 376 C21H23ClFNO2

© 2019 AB Sciex
Measuring Mass - Mass-to-Charge

Calculating m/z of Tryptophan

H O H

N CH C O

H CH2

HN

Neutral Molecule: M

M = 204
Charge (n) = +1 Charge (n) = -1
m/z = [m+n] / n m/z = [m-n] / n
= (204+1) / 1 = (204-1) / 1
= 205 = 203

© 2019 AB Sciex
Measuring Mass - Charge State

Determining Charge State - 1 Proton

Monoisotopic 1 13C 2 13C

1000 + 1 1001 + 1 1002 + 1
1 1 1

= 1001.00 = 1002.00 = 1003.00

[M + n(H+)]
1001
1002
n

M = Neutral mass
n = number of protons
1003
H+ = mass of a proton

• Note that the D mass between

isotopes is 1 Da

© 2019 AB Sciex
Measuring Mass - Charge State

Determining Charge State - 2 Proton

Monoisotopic 1 13C 2 13C

1000 + 2 1001 + 2 1002 + 2
2 2 2

= 501.00 = 501.5 = 502.0

[M + n(H+)]
501.1
501.5
n

M = Neutral mass
n = number of protons
502
H+ = mass of a proton

• Note that the D mass between

isotopes is 0.5 Da

© 2019 AB Sciex
Measuring Mass - Isotopes

Most Elements Have More than One Isotope

• Most carbon atoms have a mass of 12 Da, but in nature 1.1% of C
atoms have an extra neutron making their mass 13 Da.

Monoisotopic mass
No 13C atoms (all 12C)
1981.84
1982.84 One 13C atom

1983.84
Two 13C atoms

Mass spectrum of a peptide with 94 carbon atoms

© 2019 AB Sciex
Mass Spectrometry Data

© 2019 AB Sciex
LC-MS Workflow

Liquid sample is Ions are detected

introduced to ion and the signal is
source. communicated to
the workstation
1 which displays
5
data as a mass
spectrum

3 4

Molecules of interest Under an electrical field and Ions can be

get converted to pressure gradient, ions move transmitted, selected,
ions through the interface or fragmented in the
quadrupoles

© 2019 AB Sciex
MS: Data

Data Types
The Mass Spectrometer displays data in two ways:
• Ion Chromatogram: signal intensity change along analysis time.
• Mass Spectrum: signal intensity of m/z ratios in a mass range at a particular
time point.

Intensity
Intensity

Time (min) m/z

© 2019 AB Sciex
MS: Data

TIC Chromatogram and Mass Spectrum

TIC
• In Analyst® software if you click
on the Chromatogram (TIC) at
any time point you will open a
Spectrum.

Spectrum Spectrum
MS at 41.8 min
MS at 33.2 min

© 2019 AB Sciex
How to we measure
a specific ion?

© 2019 AB Sciex
LC-MS Workflow

Liquid sample is
introduced to ion
source.

3 4

Molecules of interest Under an electrical field and Ions can be

get converted to pressure gradient, ions move transmitted, selected,
ions through the interface or fragmented in the
quadrupoles

© 2019 AB Sciex
MS: Quadrupole Theory

What is a quadrupole?
• Quadrupole MS systems are mass filters
• Only the ion of interest is allowed to reach the detector
• Ions are separated based on m/z ratio

© 2019 AB Sciex
MS: Quadrupole Theory

What is a quadrupole?

Q0

Ions

Q1 Q2 Q3

Orifice

© 2019 AB Sciex
MS: Quadrupole Theory

How Ions travel down the Ion Path

• RF causes ions to spiral* through quadrupole.
To Detector

B A

A
A B B
B
A

*Ions travel a more complex pathway than shown. The

spiral is shown to show the simplest theoretical ion path
RF only-A rods & B rods
are 180° out of phase
© 2019 AB Sciex
Sample Ionization

© 2019 AB Sciex
LC-MS Workflow

Liquid sample is Ions are detected

introduced to ion and the signal is
source. communicated to
the workstation
1 which displays
5
data as a mass
spectrum

3 4

Molecules of interest Under an electrical field and Ions can be

get converted to pressure gradient, ions move transmitted, selected,
ions through the interface or fragmented in the
quadrupoles

© 2019 AB Sciex
MS: The Source

The IonDrive™ Turbo V Source

Ionization probe

Heated gas

© 2019 AB Sciex
Soft Ionization techniques for LC-MS

Atmospheric pressure ionization

ESI
(Electrospray Ionization)
• Turbo V™ Ion Source
• Ionizes compounds in the liquid phase

APCI
(Atmospheric Pressure Chemical Ionization)
• Turbo V™ Ion Source
• Uses a corona discharge needle to ionize compounds in the gas
phase

APPI*
(Atmospheric Pressure Photo-Ionization)
• Separate Photospray® Ion Source
• Uses a dopant and UV lamp to ionize compounds in the gas phase

© 2019 AB Sciex
General Source Usage
Which ionization technique to use?

Electrospray
(ESI)

APPI

© 2019 AB Sciex
MS: The Source

APCI setup ESI setup

• Turbo VTM source has capability of

ESI and APCI by switching probes
without the use of tools

APCI Probe • Simple hand or screwdriver

adjustable corona discharge needle
knob
ESI Probe • For APCI, corona discharge needle
is rotated in line with ceramic heater

© 2019 AB Sciex
Sample Ionization - Electrospray Ionization
LC Flow

Ion Spray Gas 1 Droplet

Voltage (IS) (GS1)
Solvent
+ Ions of Interest
Gas 2 (GS2)
& Heat (TEM) Curtain Plate
Declustering
Potential (DP)

+ +
+ - + + + + +
+ + +++-+++ ++
-+
+
+ ++ ++
+
+
Orifice

Curtain Gas
3. Ionic (CUR)
1. Formation 2. 4. Ions Enter
of Charged Evaporation Repulsion Ions Mass
Droplets Leave Droplet Analyzer

© 2019 AB Sciex
SCIEX Curtain Gas™ Interface

Curtain Plate Orifice Plate

• Robust operation
‒ Protects against dirty
matrix
Matrix
+ + ‒ Less cleaning
+ + + ++ + +++
Matrix
++++ QJet® Ion
+ +Matrix ‒ All parts removable
+ Guide
Matrix without tools
Matrix

LC
Curtain Plate
Orifice Assembly

Source ESI
Curtain
Gas
Exhaust Curtain Gas Q0 (Vacuum)

51 © 2020 DH Tech. Dev. Pte. Ltd.

Technology At a Glance - Turbo V™ Source

FOR ROBUSTNESS AND RUGGEDNESS

‒ Supports heated-ESI and APCI

‒ Sprayer has fixed distance
relative to the orifice
‒ Drying gas heaters (x2)
orthogonal to electrode

Achieve consistent results over long runs of dirty samples

MS: Sample Ionisation

Curtain Plate and Orifice Plate

• Ions are focused into skimmer by shape design of interface

• Durable, chemically inert ceramic interface
• Interface is heated and has integrated gas connections
QJet™ Ion Guide

Curtain
Plate

Interface

© 2019 AB Sciex
The IonDrive™ TurboV™ Source

Atmospheric Pressure Chemical Ionisation (APCI)

• Turbo V™ Source can perform ESI and APCI
by switching probes without tools
• Simple, adjustable corona discharge needle knob APCI Probe

ESI Probe

APCI Knob

APCI Needle
© 2019 AB Sciex
 The IonDrive™ TurboV™ Source

Atmospheric Pressure Chemical Ionisation (APCI)

Corona discharge
needle LC Flow O2 or N2
Solvent molecule
Heated
Nebulizer Ion of interest

Gas 1 Curtain Plate Curtain Gas

(GS1) (CUR)

+.
+
+ + + + +
+. +. +. +. +
+ +
+ + +
+. +. +.

Orifice
(DP)
1.Molecules 2. Corona 3. N2 or O2 4. Vaporized,
converted to discharge pass charge charged solvent 5. Ions enter
gas needle ionizes to vaporized passes charge mass analyzer
N2 or O2 in solvent to analyte aided by DP
source
© 2019 AB Sciex
The IonDrive™ TurboV™ Source

Atmospheric Pressure Photoionisation

LC Gas 1
Flow (GS1)
+ Dopant
+ Compound
Heated Curtain Plate
Nebulizer Curtain Gas
(CUR)

Krypton Lamp
+
+
+
+ . ..
+ ++ + +
+. + +
1. Molecules in
gas phase +

2. UV lamp Orifice
reacts with (DP)
dopant
3. Dopant 4. Ions enter
passes charge Mass Analyzer
to compound Aided by DP
© 2019 AB Sciex
APPI Source with Heated Nebulizer Guide

Overview
• Suitable for non-ionic, non-polar, thermally stable
compounds

Probe Temperature
• The process of distillation limits the actual temperature
experienced by analyte (150-200 ºC)
• Heating promotes vaporization

Dopant
• Requires dopant (usually toluene or acetone) post-column
and UV photons to induce ionization

Additional Parameters
• Operating range similar to that of APCI

© 2019 AB Sciex
Source Comparison

Electrospray APCI & APPI

• Applicable to a wide range of • Low polarity molecules can be ionized
analytes
• Wider dynamic range than ESI
ADVANTAGES

• Minimal fragmentation of
molecular ion • Mass sensitive detection dependency

• High molecular weights can be • Higher buffer concentration allowed

analyzed (multiple charging)

• High sensitivity
• Ionization Suppression: • Analytes must be thermally stable
DISADVANTAGES

Competition for the ions to leave

the droplet • Analytes must be volatile

• Mobile phase composition can • Dopant needed (APPI only)

affect sensitivity

© 2019 AB Sciex
 MS: Sample Ionisation

How do ions travel?

Ions travel down the ion path due to a pressure and DC voltage gradient.
Q3

Positive
QJet® II
Ion Guide
Q2
LINAC®
Ions
Collision Cell

Orifice Q0 Q1

Orifice (DP)
= 30 V Q0 =
R01 =
-10 V R02 =
-11 V
-30 V RO3 = -33 V
© 2019 AB Sciex
Mass Spectrometry:
Scan Types

© 2019 AB Sciex
Inside the MS: Tandem Mass Spectrometry

Tandem Mass Spectrometry (MS/MS)

1st set of resolving Collision Cell (q2) 2nd set of resolving

quadrupole rods (Q1) quadrupole rods (Q3)

© 2019 AB Sciex
MS: Scan Types

MS Scans MS/MS Scans

• Q1 and Q3 full range scans
• Q1 and Q3 multiple ion scans
• Product ion scan
• Precursor ion scan
• Neutral loss scan
• Multiple Reaction Monitoring
(MRM)

Q1 q2 Q3 Q1 q2 Q3

A
A X
B
B Y
C
C

© 2019 AB Sciex
MS Scan Types

MS Scan Types
• MS Scan Range (Q1 or Q3):
‒ Full range - Provides information for

intensity
every requested mass in the scan range
‒ Center Width - A narrow range scan to
obtain fast results for specific masses.
This is used for calibration and to
confirm compound characteristics 200 600 m/z

• Multiple Ions (Q1 or Q3 MI):

‒ A scan using one quadrupole as the

intensity
mass filter for specific masses
‒ Can be used for MS quantitation in
addition to MS compound parameter
optimization
200 600 m/z

© 2019 AB Sciex
MS Scan Types

The Ion Path - Reference Picture

Q1 q2 Q3

Q1 Q2 Q3
Q3

q2

Q1

© 2019 AB Sciex
MS Scan Types

Q1 and Q3 MS Scans - Start/Stop Mode

325 471 800

intensity
Q3 Q3 50 1250

q2 q2

Q1 Q1

© 2019 AB Sciex
MS Scan Types

Q1 and Q3 MS Scans - Multiple Ion Mode

intensity
Q3 Q3
325 471

q2 q2

Q1 Q1

© 2019 AB Sciex
MS/MS Quadrupole
Scan Types

© 2019 AB Sciex
MS/MS Scan Types

Fragmentation
When ions fragment:
• One or more fragment
ion(s) retain a charge
(Product Ions).
• Part of the fragment has
no charge (Neutral).
• The mass spectrometer
can only control charged
species.
Positive mode: [Precursor]+ Product+ & Neutral

Negative mode: [Precursor]- Product- & Neutral

© 2019 AB Sciex
MS/MS Scan Types

MS Scans MS/MS Scans

• Q1 and Q3 full range scans
• Q1 and Q3 multiple ion scans
• Product ion scan
• Precursor ion scan
• Neutral loss scan
• Multiple Reaction Monitoring
(MRM)

Q1 q2 Q3 Q1 q2 Q3

A
A X
B
B Y
C
C

© 2019 AB Sciex
MS: MS/MS Scan Types

Which scan type to use?

© 2019 AB Sciex
MS: MS/MS Scan Types

Product Ion Scan

• Utilized for developing MRM

transitions.
• Need to know the Q1 m/z for the
compound of interest.

© 2019 AB Sciex
MS: MS/MS Scan Types

Product Ion Scan

• Q1: Precursor ion Q3
selected
Q3

156.1
• q2: Fragmentation of 108.1
precursor occurs 92.1

• Q3: Product Ions q2

scanned

311.1

Q1
© 2019 AB Sciex
Product Ion Scan (MS2)
Reserpine (26 MCA @ different CES)

© 2019 AB Sciex
MS: MS/MS Scan Types

Precursor Ion Scan

• To determine the “origin” of a

particular product ion created in
the collision cell.
• This scan type is often used for
drug metabolite identification
(common product ion observed
in the metabolites) along with
identification of post translational
modifications in proteomics and
protein chemistry.
• Must know the Q3 m/z for the
fragment of interest.

© 2019 AB Sciex
MS/MS Scan Types

Precursor Ion Scan

• Precursor ions scanned in Q1.
• Fragmentation of precursor
Q3
occurs in q2.

156.1
• Product ion selected in Q3.

311.1
279.1
251.1

© 2019 AB Sciex
MS: MS/MS Scan Types

Neutral Loss Scan

• To determine classes of
compounds with common
fragment ions created in the
collision cell.
• This scan type is often used for
drug metabolite identification
(common fragmentation pattern
observed in the metabolites) along
with identification of post
translational modifications in
proteomics and protein chemistry.
• Must know the Q1/Q3 offset for the
neutral loss fragment of interest.

© 2019 AB Sciex
MS/MS Scan Types

Neutral Loss Scan

• Precursor ions scanned in Q1
• Fragmentation of precursor
Q3 occurs in q2
311.1-155=
156.1
• Product ion scanned in Q3
279.1-155=
124.1

311.1
279.1

© 2019 AB Sciex
MS: MS/MS Scan Types

MRM (Multiple Reaction Monitoring)

Utilizing a MRM ion pair to quantify the analyte concentration of an unknown
sample against a known concentration calibration curve through regression
analysis.

Q1 Q3
251.1 156.1

© 2019 AB Sciex
MS: MS/MS Scan Types

Multiple Reaction Monitoring (MRM)

Q3
Q3

• Q1: Precursor ion 279.1/ 156.1

selected
251.1/ 108.1

• q2: Fragmentation of
precursor occurs

• Q3: Product Ions 279.1 q2

selected 251.1

Q1

© 2019 AB Sciex
MS: MS/MS Scan Types

Multiple Reaction Monitoring (MRM)

Q1: Precursor ion q2: Fragmentation of Q3: Product Ions

selected precursor occurs selected

© 2019 AB Sciex
MS: MS/MS Scan Types

Multiple Reaction Monitoring (MRM)

© 2019 AB Sciex
Quadrupole Scan Types
Summary
 Q1/Q3 MS: Information about a predefined masse range

 SIM (Q1/Q3 MI): Identification of molecular ions

 MS2: Structural information, library search

 MRM: Increased selectivity by “SIM²”, quantitation

 Prec/NL: increased selectivity, screening for defined analytes

(e.g. metabolism studies)

© 2019 AB Sciex
QTRAP® Ready System: Two
Instruments in One

SCIEX 7500 System QTRAP® System Radial Trapping

Axial Axial
Trapping Trapping

Radial Trapping Exit Lens

with Grid

• MRM with Scheduled MRM™ Algorithm
• Q1 MS or Q3 MS scan
• Product Ion (MS/MS) scan
• Precursor Ion scan
• Neutral Loss scan
• MRM with Scheduled MRM™ Algorithm
• Q1 MS or Q3 MS scan
• Product Ion (MS/MS) scan
• Precursor Ion scan
• Neutral Loss scan
• Enhanced MS (EMS) scan
Enhanced
• Enhanced Product Ion (EPI) scan
functionality on
• Enhanced Resolution (ER) scan QTRAP system
• MS3 scan
• MRM3 scan

97 © 2021 DH Tech. Dev. Pte. Ltd.

For Research Use Only. Not for use in diagnostic procedures.
AB Sciex is doing business as SCIEX.
© 2019 AB Sciex. The trademarks mentioned herein are
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© 2019 AB Sciex