LC/MS/MS PARAMETER

MOBILE PHASE
• Aditif:
• Acetic acid at 0.1 - 1.0 % v/v
• Formic acid at 0.1% v/v
• Ammonium formate (2-10 mM optimum)
• Ammonium acetate (2-10 mM optimum)
• TFA at up to 0.1% for positive ion mode

• Note : do not used phosphates buffer

• Mobile Phase A - Methanol + 10 mM ammonium acetate
Add 10 mL 1 M ammonium acetate stock solution to 990 mL
• acetonitrile. Mobile Phase B - Water + 10 mM ammonium
acetate
Add 10 mL 1 M ammonium acetate stock solution to 990 mL water.
GRADIENT SYSTEM ISOCRATIC

• Flow rate 0.2 – 0.6 mL/min HPLC Column

• Flow rate 0.1 - 0.3 ml/min UPLC Column

%B %B

80 80

20 20

Waktu (menit) Waktu (menit)

PT. Laborindo Sarana/Theodora Rahardja/2010

SOURCE PARAMETERS
SOURCE PARAMETERS

• IonSpray Voltage (IS) [5500]

• The voltage applied between the needle and orifice plate that “ionizes” and
nebulizes the liquid flow. Polarity determines what type of ions will reach
MS. In positive mode typically 4000 and 5500V; In negative mode –3000 to
–4500V.
• Ion Source Gas 1 (GS1) [50]
• The nebulizer gas pressure. Facilitates droplet formation. Higher flow, higher
GS1.
SOURCE PARAMETERS

• Temperature (TEM) [500]

• The temperature of the heater gas (“the hairdryer”). It promotes desolvation. The setting
is optimized based on mobile phase flow rate and composition.
• Ion Source Gas 2 (GS2) [60]
• The heater gas pressure. Aids in solvent evaporation, increasing ion efficiency. Heated gas
stream intersects nebulized liquid stream at about 90o right in front of the curtain plate.
• Higher liquid flow, and/or higher aqueous mobile phase composition, higher TEM and GS2
required. Needs to be optimized.
SOURCE PARAMETERS

• Curtain Gas (CUR) [35]

• High purity N2 that flows between the orifice and the curtain plate. It
repulses large droplets and neutrals keeping the Q0 clean.

• Interface Heater (ihe) [ON]

• Orifice plate heater. I am sure it is important, but I cannot tell you why.
PARAMETER SETTING
Q0 Q1 Q2 Q3

DP EP CEP CXP
CE CEM

• Most potentials are relative to the entrance potential (EP).

• As ions move from the source to the detector they see
increasingly negative volatage.
PARAMETER SETTING

• Declustering Potential (DP) [45]*

• The voltage applied to the orifice plate.
• Used to break up ion clusters e.g.( [M+H3O+]+) and reduce chemical noise (increase sensitivity).
• HOWEVER high DP values can induce fragmentation prior to mass analysis. Generally
called “In source CID”. Great for LC/MS. Bad for LC/MS/MS.
• Entrance Potential (EP) [10]*
• The voltage between the skimmer (ground) and the entrance to Q0. Typically set to -10V in
positive mode.
• Collision Cell Entrance Potential (CEP) [10]
• The potential difference between Q0 and IQ2.
• Facilitates ion transmission to the collision cell.
PARAMETER SETTING

• Collision Energy (CE) [20 – 35 – 50 ]*

• Greater CE is usually structurally elucidating unless so high it obliterates the parent
molecule into small common mass fragments.
• Collision Cell Exit Potential (CXP) [4]
• The potential difference between Q2 and IQ3.
• Always 4V.
MASS CALIBRATION IN QUADRUPOLE MODE
PERFORM MASS CALIBRATION IN QUADRUPOLE MODE
TEST RESERPINE
CONSIDER THE AVERAGE FOOD TESTING WORKFLOW – AND ALL OF
THE QUESTIONS/CONCERNS/HEADACHES THAT COME ALONG WITH
IT…
LC
Extraction Clean-up MS analysis Identification Quantitation
separation

What is this Should I do

Am I certain of the ‘positives’?
matrix? a clean-up? Are my
Do I see
compounds
everything? Have I missed anything?
Did I extract Am I losing ionizing?
everything? compounds
of interest? Good peak Are matrix interferences
Do I have
shapes? complicating results?
Are the proper
compounds sensitivity?
stable? Is my quantitation accurate?
Matrix effects?
Are my LOQs acceptable?
BASIC LC-MS/MS
 APPLICATION LCMSMS
PROTEOMICS/PROTEIN FOOD SAFETY AND
ENVIRONMENTAL
ANALYSIS ANALYSIS

PHARMA & NATURAL CLINICAL RESEARCH AND

PRODUCT RESEARCH FORENSIC TOXICOLOGY
LCMSMS Solution for Food testing

Multi-contaminant testing
- Mycotoxins
- Polycyclic Aromatic Hydrocarbons
- Phthalates
- Food Additives
- Pesticides
- Fungicides
- Melamine
Ingredient analysis
Migrations
Halal Food Testing
Veterinary Drugs
Water Quality Testing
Vitamins Analysis
PHARMACEUTICAL APPLICATIONS

• Pharmaceutical
• BA/BE
• Impurities Profiling
• Drugs Metabolism & Pharmacokinetic (DMPK)
• ADME/Toxic

• Natural Product Research

• Natural Products profiling
• General Unknown Screening
• Biopharmaceutical
• Biosimilar
OMICS RESEARCH
• Proteomics
• Protein identification
• Protein Characterization
• Protein Quantification
• Biomarkers Discovery, verification and validation

• Metabolomics
• Metabolites profiling and quantification
• Lipidomics
• Lipid profiling and identification
• Lipid quantification
LC-MS/MS SYSTEM

Mass
HPLC
Spectrometer
Nitrogen
Generator
 Chromatographic separation with UV
detector
• LC separations occur HPLC Column

based on differential
F E D C BA
partitioning of analyte in F E D
F E D
C BA
C BA
the stationary phase
Injection UV
• Pressure-driven bulk flow Plug Detection
rates are typically at 50-
1000 mL/min AB C
DE
• Separation efficiency is F

Intensity
high

Retention time

MASS Detection
SPECTROMETER differentiates RT
UV
Detecor
THE FIRST RULE OF CHEMISTRY THE BASIS
OF MOST LIQUID CHROMATOGRAPHY
Chromatography is a method in which the components of a mixture
are separated on an adsorbent column in a flowing system.
• Components of a mixture are carried through a stationary phase by the
flow of a mobile phase.
• Separations are based on differences in migration rates among the
sample components.
The smaller the affinity a molecule has for the stationary phase, the shorter the time spent in
a column.
Stationary phase

Mobile phase
 Solvent

A+B

Packed A
B
column

A
B

detector A B

t0 t1 t2 t3 t4
Signal

A B
Time, t
WHAT’S THE MASS SPECTROMETRY & HOW DOES
IT WORK?

Ionization/ desorption Mass Spectrometer

Ion Detector
MS Analyzer
Ion Source
Form ions
(charged molecules) Sort Ions by MW to charge ratio (m/z) Detect ions

100
100

75
75

50
50
• Solid
25
• Liquid
25

Inlet
• Vapor 00
1330
1330 1340
1340 1350
1350

• HPLC
Mass Spectrum
Sample Introduction •Quantitation
Data Analysis •Structure elucidation
Method to vaporize sample
Bioinformatics
 TURBO V ION SOURCE
IONIZATION TECHNIQUES TO CONVERT MOLECULES TO IONS
WHAT IS A MASS SPECTROMETER?

A machine that measures mass

(weight of a molecule)

For example: common sugar

C12H22O11 Carbon (12 x 12)
+
Hydrogen (1 x 22)
= = + = 342.30 g/mol
Oxygen (16 x 11)

No of Carbons atoms = 12
Sucrose No of Hydrogen atoms = 22
No of Oxygen atoms = 11
MASS SPECTROMETRY
• MS measures the “molecular weight” of molecules.
• MS detects and separates ions from each other based on their mass to charge ratio
(m/z).

• For e.g.
Melamine C3H6N6
Exact/Monoisotopic Mass: 126.0654
[M+H]+ = 127.1
1
Molecular ion mass = (M + Z) / Z

where M: monoisotopic mass

Z: charge
API LC – MS/MS ION SOURCES
Two main types in use today:
1. Electro Spray Ionization (ESI)
2. Atmospheric Chemical Pressure Ionization (APCI)

optional
3. PhotoSprayTM – (APPI) atmospheric pressure photo ionization
4. MALDI "Matrix Assisted Laser Desorption/Ionization”
ELECTRO SPRAY IONIZATION (ESI)

IonSpray:
- Electrospray ionization (ESI)
- Soft ionization
- Gentlest ionization technique
- Applicable to polar and ionic substances
ATMOSPHERIC CHEMICAL PRESSURE IONIZATION
(APCI)

Heated Nebulizer: - Atmospheric Pressure Chemical Ionization (APCI)

- corona discharge
- polar to non-polar thermally stable compounds
Atmospheric Pressure Photon Ionization (APPI)

• Atmospheric Pressure Photon Ionization (APPI)

 Analytes evaporated first followed by ion-molecule reaction in the gas
phase triggered by the photon ionization
 Applicable to most non-polar compounds such as steroids, vitamins, PAHs,
etc
 Provides greater sensitivity for some compounds
ION SOURCE PROBE
 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

Fragmentation

HPLC Column Ion source Mass analyzer Detector

 MASS FILTERS MODE

m2 m1
m4 m3 m2 m1
m4 m3

mass scanning mode

m2 m1
m2 m2 m2 m2
m4 m3

single mass filtering/selected mode

MS SCAN MODES

Available Scan Modes

• Full Scan
• Provides information of all ions created during ionization
• Selected Ion Monitoring (SIM) / Q1 Multiple ion scan
• Monitors for specific ions created during ionization

m1

m3

m4

Full Scan Selected ion Scan

TRIPLE QUADRUPOLE SCAN MODES
MS Scan Modes
• Q1 or Q3 scan
• Standard MS survey scan (Provides information of all ions created during
ionization)
• SIM
• Monitors for specific ions created during ionization
MS/MS Scan Modes
• Product Ion
• Provides structural information on fragments of the original Ion
• Precursor Ion
• Provides structural information on the source Ion(s) for a specific Product Ion
• Neutral Loss
• Provides compound class specificity
• MRM
• Used for Quantitation. Since only Ions of interest are monitored this scan mode is
very specific and sensitive
PRODUCT ION SCAN
• Q1 is fixed on the precursor
ion (after identification of the
exact m/z of the precursor
using a Q1 scan)
• The precursor ion is sent into
the collision cell (Q2) and
fragmented
• Q3 scans a given mass range
to look for product ions
• Used to gather structural
information and for
identification of product ions
• Second step to developing a
Precursor Ion Fragmentation Product Ion
Selected (CAD) Scanned quantitative method
MSMS FRAGMENTATION OF MELAMINE

H
+ NH 2 NH 2 N NH 2 m/z = 85
Daughter ion
N N

NH 2 N NH 2

m/z = 127

Parent ion
 MSMS FRAGMENTATION OF MELAMINE
N

H
+ NH 2 NH 2 N NH 2 m/z = 85

N
N N
C
NH 2 N m/z = 68
NH 2 N NH 2

m/z = 127
 MSMS FRAGMENTATION OF MELAMINE
N

H
+ NH 2 NH 2 N NH 2 m/z = 85

N
N N
C
NH 2 N m/z = 68
NH 2 N NH 2

NH 2
m/z = 127
m/z = 43
N

CE (Collusion Energy) MRM 1 :127/85

MRM 2 :127/68
MRM 3 :127/43
 MASS SPECTRUM OF MELAMINE (MSMS
FRAGMENTATION)
+MS2 (127.20) CE (35): 60 MCA scans from Sample 2 (Melamine_PI_MS2) of Melamine_tune.wiff (Turbo Spray), Smoothed Max. 8.9e5 cps.

85.0
8.9e5

8.5e5
N

C
8.0e5
NH 2 N N
68.1
7.5e5
NH 2 N NH 2
7.0e5

6.5e5

6.0e5

5.5e5
In te n s ity , c p s

5.0e5

4.5e5

4.0e5
NH 2
+H
NH 2
3.5e5

3.0e5 N N N
43.1
2.5e5

2.0e5 NH 2 N NH 2

1.5e5 127.2

1.0e5
60.0
41.0 57.0
5.0e4 67.0
55.0 79.0 80.9 110.3
65.1
0.0
40 45 50 55 60 65 70 75 80 85 90 95 100 105 110 115 120 125 130 135 140 145 150
m/z, Da
IDENTIFICATION OF EURYCOMANONE IN TONGKAT ALI
EXTRACT

MRM

20eV
PI

35eV
PI

PI 50eV
CURRENT EU CRITERIA FOR TARGET CONFIRMATION
Guideline 2002/657/EG

SIM and MRM (4 identification points)

MS precursor 1.0
2 MRMs = 4 points
MSn product 1.5

Other guidelines
Full scan spectra (ions > 10% of base peak, mass spectral libraries)
High resolution MS (>10,000)
Introduction to MS-driven Quantitation
What is Multiple Reaction Monitoring (MRM) ?

Multiple Reaction Monitoring (MRM) provides the highest sensitivity and

specificity for quantitation in complex mixtures
MULTIPLE REACTION MONITORING (MRM)

In MRM mode, Q1 and

Q3 width is set to 0
Many precursor to
product ion pairs can be
monitored (A-B, A’-B’,
A”-B”, etc.)
MRM analysis is the
best way to maximize
signal intensity of
product ions
Precursor ion Fragmentation Product ion MRM used primarily for
Selected (CAD) Selected quantitation studies
 MRM CHROMATOGRAM OF MELAMINE
SAME RT FOR SAME COMPOUND
SAME PARENT WITH 3 DIFFERENT DAUGHTERS

XIC of +MRM (3 pairs): 127.000/85.300 Da from Sample 3 (40ppb std_ACN) of Std inertstil column batch .wiff (Turbo Spray) Max. 1.7e5 cps.

5.61
1.7e5

1.6e5

1.5e5 127/85
1.4e5

1.3e5

1.2e5

1.1e5

1.0e5
In te n s ity , c p s

9.0e4 127/68
8.0e4

7.0e4

6.0e4

5.0e4

4.0e4
127/43
3.0e4

2.0e4

1.0e4

0.0
3.8 4.0 4.2 4.4 4.6 4.8 5.0 5.2 5.4 5.6 5.8 6.0 6.2 6.4 6.6 6.8 7.0 7.2 7.4 7.6 7.8
Time, min
MODE OF OPERATION -QTRAP TECHNOLOGY

Triple Quadrupole Mode of Operation TripleQ

1. Q1 & Q3 mass filter for Neutral loss scan 

2. Q1 & Q3 mass filter for Precursor ion scan 

3. Q2 Collision cell for true high quality MS/MS spectra for


structural information

4. MRM scan MS/MS quantification (without library matching) 

IMPORTANCE OF DWELL TIME IN MRM
MODE
SIGNAL TO NOISE XIC of +MRM (5 pairs): 475.170/283.100 Da from Sample 12 (sildenafil 100 ppb) of presisi.wiff (Turbo Spray)

1.4e4
0.67
Max. 1.4e4 cps.

S/N = 2596.3
1.3e4
Peak Int.(Subt.)=1.4e+4
1.2e4
1xStd.Dev.(Noise)=5.4e+0
1.1e4

1.0e4

9000.0

8000.0
LOD Minimal 3 x SN
I n t e n s it y , c p s

7000.0
LOQ Minimal 10 x SN
6000.0

5000.0

4000.0

3000.0

2000.0

1000.0

! - Noise - !
0.0
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0
Time, min
 CROSS TALK
LINAC COLLISION CELL TECHNOLOGY ELIMINATES CROSS-TALK
XIC of +MRM (4 pairs): for 188.2 / 91.1 amu from Seleg 2500pg No-LINAC 1.04e6 cps XIC of +MRM (4 pairs): for 188.2 / 91.1 amu from Seleg 2500pg w LINAC 3.76e6 cps
100
25 ng Selegiline 3e6
25 ng Selegiline
80

Intensity, cps
60
% Intensity

2e6

40
1e6
20

0.5 1.0 1.5 2.0

0.5 1.0 1.5 2.0
Time, min
Time, min

XIC of +MRM (4 pairs): for 136.1 / 91.1 amu from Seleg 2500pg w LINAC 8.66e3 cps
XIC of +MRM (4 pairs): for 136.1 / 91.1 amu from Seleg 2500pg No-LINAC 3.67e4 cps
100
Dw e l l 1 0 0 m s e c 8000
No cross-talk visible with LINAC
80
False Amphetamine 6000
signal detected

Intensity, cps
60
% Intensity

40 without LINAC 4000

2000
20

0.5 1.0 1.5 2.0 0.5 1.0 1.5 2.0

Time, min Time, min

• Selegeline MRM: 188.2 amu > 91.1 amu, Amphetamine MRM: 136.1 amu > 91.1 amu

• 25 ng Selegeline injected, but NO amphetamine

 Thank you

61
CRITICAL POINTS ANALYSIS LC-
MS/MS
 CONSIDER THE AVERAGE FOOD TESTING WORKFLOW – AND ALL OF
THE QUESTIONS/CONCERNS/HEADACHES THAT COME ALONG WITH
IT…
LC
Extraction Clean-up MS analysis Identification Quantitation
separation

Matrix Sample Should I do

Am I certain of the ‘positives’?
Characteristic a clean-up? Are my
Analyt Do I see
compounds
everything? Have I missed anything?
Am I losing ionizing?
compounds
of interest? Good peak Are matrix interferences
Do I have
shapes? complicating results?
proper
sensitivity?
Is my quantitation accurate?
Matrix effects?
Are my LOQs acceptable?
Benefits of QTRAP® – Overcome matrix ambiguity

Example

Q: ARE MATRIX INTERFERENCES COMPLICATING MY RESULTS?

A. Enhanced Product Ion (EPI) scanning with QTRAP® is
basically like having more than 4 MRMs for any detected
compounds in your sample.
XIC of +MRM (202 pairs): 202.0/1... Max. 4.0e4 cps. XIC of +MRM (202 pairs): 297.0/... Max. 7300.0 cps.

6.4 9.1
4.0e4 10ppb Imazalil7000
3.5e4
6000
3.0e4 MRM ratio =
In ten sity, cp s

In ten sity, cp s
5000
2.5e4 0.63  0.093
4000
2.0e4
3000
1.5e4

1.0e4 2000

5000.0 1000

0.0 0
3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 7 8 9 10 11 12
Time, min Time, min
XIC of +MRM (202 pairs): 202.0/1... Max. 5.8e4 cps. XIC of +MRM (202 pairs): 297.0/... Max. 1000.0 cps.

6.5 9.3
5.8e4 1000
Imazalil in
5.0e4
sample?? 800

(10x diluted
?
In te nsity, cp s

In te nsity, cp s

4.0e4
600
3.0e4 extract)
400
2.0e4
MRM ratio =
200
1.0e4 0.50
0.0 0
4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 7 8 9 10 11 12
Time, min Time, min
Benefits of QTRAP® – Overcome matrix ambiguity

Example

Q: ARE MATRIX INTERFERENCES COMPLICATING MY RESULTS?

A. Enhanced Product Ion (EPI) scanning with QTRAP® is
basically like having more than 4 MRMs for any detected
compounds in your sample.
MS/MS Library fit = 90.4%
XIC of +MRM (202 pairs): 202.0/1... Max. 4.0e4 cps. XIC of +MRM (202 pairs): 297.0/... Max. 7300.0 cps.
XIC of +MRM (101 pairs): Exp 1, ... Max. 1.1e5 cps. XIC of +MRM (101 pairs): Exp 1... Max. 2500.0 cps.
6.4 9.1
4.0e4 10ppb Imazalil7000 1.11e5
6.6
2500
9.4

1.00e5
3.5e4


2000
6000 8.00e4

MRM ratio =

I n t e n s it y , c p s

I n t e n s it y , c p s
3.0e4 1500
In ten sity, cp s

In ten sity, cp s
5000 6.00e4

2.5e4 0.63  0.093 4.00e4

1000
4000
2.0e4 2.00e4 500

3000
1.5e4 0.00
4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5
0
7 8 9 10 11 12
Time, min Time, min
1.0e4 2000 +EPI (202.00) Charge (+0) CE (3... Max. 2.9e6 cps. +EPI (297.00) Charge (+0) CE (3... Max. 8.0e5 cps.

297.2
5000.0 1000 4.0e6
8.0e5
159.1
7.0e5
3.5e6
0.0 0 3.0e6 202.2 6.0e5
3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 7 8 9 10 11 12

I n t e n s it y , c p s

I n t e n s it y , c p s
5.0e5
2.5e6
Time, min Time, min 175.2
2.0e6 4.0e5
XIC of +MRM (202 pairs): 202.0/1... Max. 5.8e4 cps. XIC of +MRM (202 pairs): 297.0/... Max. 1000.0 cps.

can
1.5e6 3.0e5 69.0

I s
173.2
6.5 9.3 2.0e5

P
1.0e6 131.2
5.8e4 1000
E
109.2 176.2201.0
Imazalil in 1.0e5 81.1
5.0e5 255.2
65.0 92.2 104.2 143.2
0.0
5.0e4 60 80 100 120 140 160 180 200 220 240 50 100 150 200 250 300 350

sample?? 800 m/z, amu m/z, amu

(10x diluted
?
In te nsity, cp s

In te nsity, cp s

4.0e4
600
3.0e4 extract) • More fragments to confirm
2.0e4
400 • Ability to compare to a library spectrum
MRM ratio =
200
1.0e4 0.50 The EPI data can help to de-complicate any
0.0
4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0
0
7 8 9 10 11 12
effects that the matrix might have on the MRM
Time, min Time, min ratio alone for better confirmation of positives.
Benefits of QTRAP® – Added confidence

Q: AM I SURE OF THE POSITIVE HITS?

A: YOUR QTRAP® CAN GIVE YOU THAT EXTRA SENSE OF

SECURITY
XIC of +MRM (150 pairs): 280.2/2...
WHEN CONFIRMING
Max. 1.7e4 cps. +EPI (280.00) CE (35) CES (15):... Max. 6.6e7 cps. MRM POSITIVES.
XIC of +MRM (150 pairs):ratio
280.2/2... would
Max. 1.7e4 cps. +EPIsuggest
(280.00) CE (35) CES (15):... this
Max. 6.6e7 cps.

1.6e4
7.7
10 ppb Metalaxyl6.6e7
160.1
sample
1.6e4
was
7.7
positive for
160.1
6.6e7Library spectrum
Standard 6.0e7
Metalaxyl
6.0e7 Metalaxyl
1.4e4 1.4e4
5.0e7 5.0e7
1.2e4 1.2e4
In te n sity, cp s

In te n sity, cp s

In te n sity, cp s

In te n sity, cp s
MRM ratio = 0.744.0e7 4.0e7
1.0e4 1.0e4
8000.0 3.0e7 145.1 192.2 RT
8000.0 was slightly off
3.0e7
(but 145.1 192.2not

6000.0
2.0e7
132.1 148.1
162.2 unusual
6000.0 in food matrices)
2.0e7
132.1 148.1
162.2
4000.0 220.3 4000.0 220.3
134.0 134.0
1.0e7 105.1 1.0e7 105.1
2000.0 6.2 248.3 280.3 2000.0 6.2 248.3 280.3
0.0
2 4 6 8 10 12 14 50 100 150 200 250 300 350 400
Am
0.0 I sure??
2 4 6 8 10 12 14 50 100 150 200 250 300 350 400
Time, min m/z, amu Time, min m/z, amu
XIC of +MRM (150 pairs): 280.2/... Max. 2100.0 cps. +EPI (280.20) Charge (+0) CE (3... Max. 3.3e5 cps. XIC of +MRM (150 pairs): 280.2/... Max. 2100.0 cps. +EPI (280.20) Charge (+0) CE (3... Max. 3.3e5 cps.

7.9 280.3 7.9 280.3

?
3.3e5 3.3e5
2000 2000
Metalaxyl
Unknown sample3.0e5

3.0e5
in sample?
2.5e5 spectrum 2.5e5
1500 (10x diluted 1500
In te n sity, cp s

In te n sity, cp s

In te n sity, cp s

In te n sity, cp s
Calculated FIT for Metalaxyl =
extract) 2.0e5 2.0e5
concentration = 23.6%
1000 220.3 1000 220.3
1.5e5 13.3μg/kg 1.5e5
192.4 192.4
500 1.0e5 148.3 500 1.0e5 148.3
MRM ratio =
5.0e4 5.0e4
0.72
134.0 202.3 134.0 202.3
0 0
2 4 6 8 10 12 14 50 100 150 200 250 300 350 400 2 4 6 8 10 12 14 50 100 150 200 250 300 350 400
Time, min m/z, amu Time, min m/z, amu
Benefits of QTRAP® – Added confidence

Q: AM I SURE OF THE POSITIVE HITS?

A: YOUR QTRAP® CAN GIVE YOU THAT EXTRA SENSE OF

SECURITY
XIC of +MRM (150 pairs): 280.2/2...
WHEN CONFIRMING
Max. 1.7e4 cps. +EPI (280.00) CE (35) CES (15):... Max. 6.6e7 cps.
POSITIVES.
XIC of +MRM (150 pairs): 280.2/2... Max. 1.7e4 cps. +EPI (280.00) CE (35) CES (15):... Max. 6.6e7 cps.

7.7 160.1 7.7 160.1

6.6e7Library spectrum
1.6e4 10 ppb Metalaxyl6.6e7 1.6e4
Standard 6.0e7 6.0e7 Metalaxyl
1.4e4 1.4e4
5.0e7 5.0e7
1.2e4 1.2e4
In te n sity, cp s

In te n sity, cp s

In te n sity, cp s

In te n sity, cp s
MRM ratio = 0.744.0e7 4.0e7
1.0e4 1.0e4
8000.0 3.0e7 145.1 192.2 8000.0 3.0e7 145.1 192.2

6000.0 132.1 148.1 6000.0 132.1 148.1

2.0e7 162.2 2.0e7 162.2
4000.0 220.3 4000.0 220.3
134.0 134.0
1.0e7 105.1 1.0e7 105.1
2000.0 6.2 248.3 280.3 2000.0 6.2 248.3 280.3
0.0 0.0
2 4 6 8 10 12 14 50 100 150 200 250 300 350 400 2 4 6 8 10 12 14 50 100 150 200 250 300 350 400
Time, min m/z, amu Time, min m/z, amu
XIC of +MRM (150 pairs): 280.2/... Max. 2100.0 cps. +EPI (280.20) Charge (+0) CE (3... Max. 3.3e5 cps. XIC of +MRM (150 pairs): 280.2/... Max. 2100.0 cps. +EPI (280.20) Charge (+0) CE (3... Max. 3.3e5 cps.

7.9 280.3 7.9 280.3

?
3.3e5 3.3e5
2000 2000
Metalaxyl
Unknown sample3.0e5

3.0e5
in sample? The EPI data can help
2.5e5 spectrum 2.5e5
1500 (10x diluted 1500 to de-complicate the
In te n sity, cp s

In te n sity, cp s

In te n sity, cp s

In te n sity, cp s
Calculated FIT for Metalaxyl =
extract) 2.0e5 2.0e5 results for better
concentration = 23.6%
1000
1.5e5
220.3
13.3μg/kg
1000
1.5e5
220.3 confirmation of
1.0e5
192.4
1.0e5
192.4
positives – or, in this
500 148.3 500 148.3
MRM ratio = case, confirmation that
5.0e4 5.0e4
0.72
134.0 202.3 134.0 202.3
this peak is not
0 0
2 4 6 8 10
Time, min
12 14 50 100 150 200 250 300 350 400
m/z, amu
2 4 6 8 10
Time, min
12 14 50 100 150 200 250 300 350 400
m/z, amu
Metalaxyl.
 Thank you

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