1. Introduction to Liquid Phase Mass Spectrometry
Andrew Baca
Business Development Specialist Environmental and Food Markets

2. Topics
Detectors
Liquid Phase
Example Applications

3. LCMSMS Detectors Triple Quadrupoles Ion Traps
HRAM Benchtop Orbitraps ™
Hybrid Orbitraps ™

4. Quadrupole vs. Ion Traps
Single Quadrupole
Ion Traps VS
Filter
Bucket

5. Single Quadrupole Principles of Operation
Full Scan (FS)
Ion Source
Detector
Diagonal line represents
the energies that are
scanned to allow one ion
at a specific mass to
charge ratio to go through
the quadrupoles at a time.
Single Ion Monitoring (SIM)
Ion Source
Detector
Straight line represents
the energy that is applied
to allow only one ion
at a specific mass to
charge ratio to go through
the quadrupoles.

6. Triple Quad MS – Principle of Operation
Quantitation of target compounds in matrix samples
Select
Fragment
Detect
Argon Collision Gas
Ion Source
Detector
Q1 selects the precursor ion
DDT ion: m/z
Q3 selects the product ion
fragments to m/z

7. Ion Trap Principle of Operation
Trap all m/z ions simultaneously.
Mass-selectively eject them.
Ion
Injection
Ion
Trapping
Mass-selective
Ejection

8. Orbitrap – Principle of Operation
Kingdon Trap 1923 z φ
Hyper-logarithmic potential distribution:
“ideal Kingdon trap” r
Characteristic frequencies:
Frequency of rotation ωφ
Frequency of radial oscillations ωr
Frequency of axial oscillations ωz
Makarov A. Anal. Chem. 2000, 72,

9. Triple Stage Quadrupole Systems
Target market:
Most sensitive system
Most demanding quantitative requirements.
TSQ Vantage
3,000 timed SRMs per LC run
EMR for peptide Quan
> 4 orders linear dynamic range
Sensitivity: ESI/APCI 500 fg injected on column s/n 500:1
TSQ Quantum
Ultra
Performance
Target market:
Routine quantitation
highly selective reaction monitoring (H-SRM) for greater analytical selectivity
Sensitivity: ESI/APCI 500 fg injected on column s/n 150:1
TSQ Quantum
Access Max
Target market:
Environmental and food safety
Entry level quantitation, best in Class, extremely good value
Customers with limited budgets
Mass range m/z – Small molecules to intact proteins
Fast +/- switching for high-speed
Sensitivity: ESI/APCI1 pg injected on column s/n 200:1
Value

10. HSRM HSRM S/N 3918 Unit Resolution S/N 289 Matrix background peak!
Chromatogram on the right (HSRM) shows improved S/N over left (unit resolution). By increasing the resolution on Q1 to 0.1 FWHM the TSQ Vantage can eliminate matrix background noise as well as interference peaks. HSRM provides enhanced selectivity for the ion of interest resulting in fewer false positives!
S/N 3918
Unit Resolution
S/N 289
Matrix peak removed by HSRM

11. Ion Trap Systems Performance Value Target market:
Velos Pro
Target market:
Peptide ID and Quan in one system
Metabolite ID and Quan in one system
Premium Ion Trap System
Mass range m/z
Worlds first quantitative Ion Trap
Worlds fastest most sensitive Ion Trap
CID, PQD, ETD, HCD for MSN
LTQ XL
Target market:
Peptide ID and Metabolite ID
Small molecule structural elucidation
Ultimate flexibility – upgrade to MALDI, ETD, Orbitrap
Performance
Mass range m/z
CID, PQD, ETD for MSN
Upgrade to Orbitrap
LCQ Fleet
Target market:
Ion trap of choice for routine analysis of complex matrices & multi-component samples
Single Quad Replacement
Ease of use for non-MS operators
Mass range m/z – Small molecules to intact proteins
CID, PQD for MSN
Value

12. Ion Trap: Data Dependent MSN
At each point in the chromatogram we obtain a unique MSN spectrum.
The high intensity ion in MS1 triggers data dependent acquisition to any desired level of MSN.
In this example, we have MS2, MS3 and up to MS4 (not shown)
The full collection of MSN spectra is referred to as a Spectral Tree.
The Spectral Tree uniquely defines the structure of a molecule – just like a fingerprint. 50
100
150
m/z 10 20 30 40 60 70 80 90
Relative Abundance
121.25
147.29
57.08
165.17
93.06
MS3
100
150
200
250
300
m/z 10 20 30 40 50 60 70 80 90
Relative Abundance
177.31
286.62
110.23
195.38
132.42
93.26
MS3
200
300
400
500
m/z 10 20 30 40 50 60 70 80 90
100
Relative Abundance
177.26
303.69
272.58
391.81
MS2
300
400
500
600
700
m/z 10 20 30 40 50 60 70 80 90
100
Relative Abundance
485.93
MS1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Time (min)

13. HRAM Benchtop Orbitrap ™ Systems
Q Exactive
Target market:
Q-TOF competitor
Little Method Development
Environmental and Food Safety
Mass Range m/z
Mass Accuracy: <1ppm
MS/MS for confirmation and ID
Mass resolution: 140,000
Performance
Exactive Plus
Target market:
TOF competitor
Multiple component screening and quantitation
Ease of use for non-MS operators – no method development
Mass Range m/z
Mass Accuracy: <1ppm
Mass Resolution: 140,000 (1.5 scan/sec); 17,500 (12 scans/sec)
Value

14. Resolution and Mass Accuracy
FWHM
10%
Mass accuracy
Before getting details in applications of Exactive for non-target screening, let’s understand two important concepts in Exactive.
Click
What is resolution? Understanding resolution in MS is just as simple as in picture.
When the resolution of a picture is low, you can not see clearly what is in the picture.
When the resolution is improved, the picture is clear, see bridge, sky.
Resolution for MS is calculated as R=m/delta m, defined as FWHM (full width at half maximum) for TOF/Orbitrap MS, or resolution at 10% valley for magnetic sector MS.
Mass accuracy is calculated as the formula. It tells how accurate the MS can measure the mass of a compound.

15. 510 Pesticides analysis in 12 Minutes

16. Hybrid Orbitrap Systems
Orbitrap Elite
Target market:
Complete Proteomics Solution
Metabolite ID
Worlds Best mass spectrometer
Mass range m/z
Velos Pro Ion Trap
CID, PQD, ETD, HCD for MSN
HR/AM >240,000 FWHM
Orbitrap Velos Pro
Performance
Target market:
Peptide ID and Metabolite ID
Small molecule structural elucidation
Ultimate flexibility – upgrade to MALDI, ETD, Orbitrap
Mass range m/z
Velos Pro Ion Trap
CID, PQD, ETD, HCD for MSN
HR/AM > FWHM
LTQ Orbitrap XL
Target market:
Amazing analytical performance at this price point
Peptide ID and Small Molecule structural elucidation
MALDI option
Mass range m/z
LTQ XL Ion Trap
CID, PQD, ETD, HCD for MSN
HR/AM > FWHM
Price

17. Example Application LC Triple Quad Vitamins
Istds
Vitamins
Niacinamide
Pyridoxine
Pantothenic Acid
Thiamine
Folic Acid
Biotin
Riboflavin

18. Example Application HRAM Benchtop Orbitrap ™ Systems
Mass Accuracy ppm
Detection from Blank
Simulation

19. After Melamine DEHP, what is the next one? ???
Is an industrial chemical
Not approved for use in food
DEHP
Plasticizer
Not approved for use in food
Most food testing labs are doing target analysis for food contaminants. Even some scientist think non-target screening methods are unnecessary. It is true that food safety testing is regulatory driven. When food products do not contain regulated contaminants higher than the MRLs, they can be distributed into market. We do need to ask: does target analysis guarantee our food safe?
Click
I believe we still have fresh memory for the melamine tainted milk scandal in 2008 that began with Sanlu Dairy”s products. This scandal made almost 300,000 children sick and caused 4 deaths. However, melamine is an industry chemical, not approved for use in food. Thus melamine was undetectable in target analysis. We did not catch the misuse of melamine until severe consequences came.