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Introduction to DART MS
Robert B. Cody JEOL USA, Inc.

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Outline

Definition of terms DART operating principle TOF mass spectrometer overview The information we obtain

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Definitions of MS terms and general concepts

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High Resolution Mass Spectrometry

We will be using exact-mass measurements to to confirm knowns and to determine elemental compositions for unknowns Resolving power defines how well the mass spectrometer can separate close peaks (interferences) The elemental composition software gives us other information for each candidate composition (e.g. unsaturation)

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Resolving Power

R=M/ M
R = Resolving Power M = m/z M = difference in mass that can be separated

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Resolving Power Defined as: FWHM (Full width at half maximum)

R=M/ M R = 5000 m/z 500 M = Peak width at half-height = 0.1

0.1

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Resolving Power Defined as: 10% Valley Definition

R=M/ M R = 500 m/z 500 and 501 can be separated at a 10% Valley M=1
500

501

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Examples for C36 H74 (m/z 506.579)

R = 500 (10% valley) Separate m/z 500 from 501

R = 5000 (10% valley) Separate m/z 500 from 500.1

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Why the definition matters

R = 500 (10% valley)

R = 500 (FWHM)

R = 5000 (FWHM)

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Mass accuracy

millimass units (0.001) or mmu

ppm = 106 * ( M / M)
parts-per-million (ppm)
Resolution (reciprocal of resolving power) Note: ppm is a m/z dependent value

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Unsaturation
(aka rings and double bonds aka double bond equivalents)
O

H+

CH3COOD = 1.5, subtract 0.5

CH3

H3C

C6H6+. D = 4.0

C3H7O+. D = 0.5, add 0.5

H3O+ D = -0.5, add 0.5

Value is calculated from elemental composition Indicates total rings, double bonds, triple bonds Exact integer (e.g. 4.0) or half-integer (3.5)

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Examples of Even-electron ions and Odd-electron ions

Protonated molecule [M+H]+ Deprotonated molecule [M-H]Chloride adduct [M+Cl]Ammoniated molecule [M+NH4]+ Fragment F+

Even-electron ions (half integer unsaturation) :

Odd-electron ions (exact integer unsaturation) :

Molecular radical cation M+. Molecular radical anion M-. Fragment F +.

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On-line Resources
DART Users Google Newsgroup
http://groups.google.com/group/dart-mass-spectrometer-users?hl=en

JEOL USA, Inc. Web Pages

http://www.jeolusa.com

IonSense Web Page

http://www.ionsense.com

Wikipedia article on DART

http://en.wikipedia.org/wiki/DART_ion_source

Proton affinities, ionization energies (NIST)

http://webbook.nist.gov/chemistry/

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DART Basic Principles

See the JEOL News Article on the AccuTOF-DART product page on www.jeolusa.com

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DART: Direct Analysis in Real Time

Operational in Jan. 2003 Patent filed in April 2003 Public disclosure, Jan. 2005 Commercial product introduced March 2005 First open-air, ambient ion source for MS

1. Cody, R. B.; Laramee, J. A. Method for atmospheric pressure ionization US Patent Number 6,949,741 issued September 27, 2005. 2. Laramee, J. A.; Cody, R. B. Method for Atmospheric Pressure Analyte Ionization US Patent Number 7,112,785 issued September 26, 2006.

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Prototype DART sources

Original prototype DART source (mid-2002)

Second DART prototype (Early 2003)

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The Whole Package:AccuTOF-DART

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Why DART?
Fast and easy way to introduce samples Minimal sample preparation for most samples Can tolerate dirty or high-concentration samples and without contamination Fast fingerprinting of materials

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Nothing comes without a price

Chromatography/MS still has advantages over DART in detection limits, selectivity and sensitivity for certain samples Not useful for large biomolecules (no good for DNA analysis, proteins) DART does not ionize metals, minerals, etc.

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DART Schematic

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DART Ionization
Penning ionization Sample ionized directly by energy transfer from metastables (M*) Proton transfer (positive ions)

M*
DART Source

1. He* ionizes atmospheric water 2. Ionized water clusters transfer proton to sample Electron capture (negative ions) 1. Penning electrons rapidly thermalized 2. Oxygen captures electrons 3. O2- ionizes sample

MS API Interface

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Penning Ionization
Metastable atoms or molecules react with analytes that posses ionization potentials less than the metastable energy, M* + S S+. + M + electron The helium 23S state has 19.8 eV of internal energy and lasts up to 8 minutes in vacuum.
Most molecules have ionization energies much lower than 19.8 eV

Proton Transfer

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He(23S) + H2O H2O+ + He(11S) + electron H2O+ + H2O H3O+ + OH H3O+ + nH2O [(H2O)n+1 H]+ [(H2O)nH]+ + M MH+ + nH2O
Metastable atoms react with atmospheric water to produce ionized water clusters Dominant reaction mechanism when helium carrier used: He(23S) energy = 19.8 eV Huge reaction cross section: 100 A2

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Typical DART Low-Mass Background

Normal DART Parameters

100
80

[(H2O)2+H]+

Rel. Abund.

60 40 20 0

NH4+

H3O+ NO+

[(H2O)3+H]+

15

20

25

30

35

40

45

50

55

m/z

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Negative Ion Formation

Electrons produced by direct or surface Penning ionization are rapidly thermalized Thermal electrons react with atmospheric oxygen and water to produce ionized clusters Oxygen/water cluster ions react with analyte molecules to produce analyte ions e-* + G e- + G* e- + O2 O2-. O2-. + S [S-H]- + OOH. O2-. + S S-. + O2 O2-. + S [S+O2]-.* + G [S+O2]-. + G*

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Typical DART Negative-Ion Low-Mass Background

O2[H2O3][H2O4][H C O]3
20 40 m /z 60

Rel . abundance

[H C O]4
80 10 0

Note the absence of nitrogen oxide ions that would be produced by electrical discharge in air. NO2- and NO3- are problematic for detection of nitro explosives and reduce anion detection sensitivity

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9 5

Example
Ascorbic acid, C6H8O6
5 2 HO

O H HO

[M+H]+
O

Rel. Abund.

100
50 0

HO

177.0410

Positive ions

9 (m a

1 2 7 1 3 0 1 3 3 1 3 6 1 3 9 1 4 2 1 4+ 1 4 8 1 5 1 1 5 4 1 5 7 1 6 0 1 6 3 1 6 6 1 6 9 1 7 2 5 2 i )n A l i s c o r b i c b A c id

[M+H-2H O]

[M+H-H2O]+

100

150

m/z
Rel. Abund.

[M-H]175.0232

100
50 0

Negative ions

100

150

m/z
Sampled directly from a melting point tube

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Notes on the AccuTOF Design and Operation

See the JEOL News Article on the AccuTOF-LC product page on www.jeolusa.com

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Types of mass spectrometers

Scanning:
magnetic sector, quadrupole and triple quadrupole

Trapped-ion:
Fourier transform, 3D ion trap, Orbitrap linear trap (used in triple quadupole MS)

Time-of-flight Hybrids

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DART can be fit on most mass spectrometer types

DART signals can be transient, so,

scanning mass specs work best with selected ion monitoring or fast scanning Selected reaction monitoring on triple quadrupole MS is good for target compound quantitation. Ion traps work, but are not a good choice for quantitative analysis Time-of-flight is fastest MS for transient signals, and gives high-resolution data for the entire mass spectrum with no sensitivity loss.

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Time of flight principle

If everyone starts at the same time and has the same kinetic energy, lighter riders will move faster
Light ions moving quickly Detector

Heavy ions moving slowly

LAlpe DHuez de Spectrometrie de Masse

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A more realistic TOF mass spectrometer

Ion source: Short burst of ions

Flight tube
High voltage to accelerate ions

Ion detector

Kinetic Energy = qE = mv2/2

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What if ions that have the same mass have slightly different energies?

Reflectron: make the more energetic ions travel further

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Reflectron Time of flight mass analyzer principle

1. Fast riders miss the turn

Me

Lance

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Reflectron Time of flight mass analyzer princip

2. Fast riders turn around; have to travel further

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Reflectron TOF

3. Fast riders start to catch up

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Reflectron TOF

Focal point

4. Fast riders catch up, will eventually pass

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Time-of-flight math

All ions fly with the same kinetic energy.

1 ( M mu ) v 2 = q e V 2
M: mass of ion [u] mu: Atom mass unit (1.6605 x 10-27 [kg/u]) q: charge number of ion V: Accelerating voltage [V] v: flight speed of ion [m/s]

e: unit electric charge (1.602 x 10-19 [C])

Flight time is inversely proportional to the square root of the mass/charge ratio.

L q tof M V

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JMS-T100LC AccuTOFTM
Ion Source Ion Transportation Analyser

Detection system To the data collection system

TMP2 RP TMP1 RP

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AccuTOFTM Ion Source

Ion Source Ion Transportation Analyser

Detection system To the data collection system

TMP2 RP TMP1 RP

Technology Transition Workshop Orthogonal ESI ion source and API interface LC Eluent Nebulizer Gas Desolvating Chamber

Desolvating Gas

Orifice2 Ion Guide

Ring Lens Orifice1 RP TMP

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Ion Source and Atmospheric Pressure Ionization (API) Interface

Orthogonal ESI
Minimize contamination into API interface

Simple API interface

Robust, few parameters, minimal maintenance

Off-axis skimmers and ring lens, bent ion guide

Keep contamination out of high-vacuum region

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AccuTOFTM Ion Transport

Ion Source Ion Transportation Analyser

Detection system To the data collection system

TMP2 RP TMP1 RP

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Ion transport region

Strong acceleration of ions only occurs in high-vacuum region

Minimize CID and scattering

Quadrupole RF ion guide focuses ions to a small spot size

Spatial focus for good resolution High-pass filter (ions greater than given m/z)

Multi-function focusing and steering lenses

Beam should be perpendicular

AccuTOFTM Analyzer
Ion Source Ion Transportation Analyser

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Detection system To the data collection system

TMP2 RP TMP1 RP
z y x

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AccuTOFTM Analyzer
z y (injection) x (reflectron)

Two-step acceleration
Spatial focusing of ion beam

Single reflectron

Energy focusing of ion beam in the x-direction Minimize ion loss

oa(Orthogonal-Acceleration)-TOF MS
Kinetic energy spread in y-direction has no effect on resolution The ions produced by the ESI ion source are used efficiently.

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Flight cycle of oa-TOF MS

1. Introduction of ion
Two kinds of ions are introduced at the same time.

Ion Source

Low mass ion High mass ion Mixture of both ions

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Flight cycle of oa-TOF MS

2. Turn on the pulser voltage
Mixture of ions at the start of flight

+ + + + + +

Ion Source

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Flight cycle of oa-TOF MS

3. Turn off the pulser voltage
continuing flight mass separation

Ion Source

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Flight cycle of oa-TOF MS

New ions are introduced in the ion acceleration part.

4. Continuing flight

Ion Source

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Flight cycle of oa-TOF MS

The ion acceleration region is filled with the new ions.

5. Low mass ion reaches detector

Ion Source

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Flight cycle of oa-TOF MS

6. High mass ion reaches detector

Ion Source

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Flight cycle of oa-TOF MS

7. The detection of all ions is completed

Ion Source

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AccuTOFTM Detection system Detection

Ion Source Ion Transportation system Analyser To the data collection system

TMP2 RP TMP1 RP

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Detector
in the atmosphere in the vacuum To impedance converter

Anode Dual MCP

a.

e-

a.

Micro-channel plate (MCP) 40mm Dual MCP Anode Combined with high voltage capacitor Patent pending

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MCP

Diameter 40mm Thickness 0.6mm I.D. of channel 10m Gap of each channel 12m

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Data collection system for oa-TOF MS

- Requirements High time resolution

m/z 609, R=6,000 Peak width: 3.5ns

Continuous data collection

High duty cycle

Real-time accumulation of mass spectrum

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Data collection system for oa-TOF MS

TDC
Super-high speed digital stop watch Measures the arrival time of ions A premise is that there are a few ions
Each ion arrives separately. Ion counting detection: signal is 0 or 1.

Continuous Averager
A signal from the detector is converted digital value by a highspeed ADC (Analog-toDigital Converter). Spectrum can be accumulated continuously in real time.

TDC (Time-to-Digital Converter)

59us Start Input High Voltage Pulser Amp Discriminator

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Time-to-Digital Converter
Stop Input

No. TOF [us] 1 29.4235 2 46.2890 ....

No. of Ions Detected in a Cycle

Histogram Memory
To Data System

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Output fromAm plifier Cycle 1 20 15 m V 10 5 0 1 5 9 13 17 21 25 29 33 37 41 45

Histgramm ory em Cycle 1 3 2 1 0 1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46

No. of Ions

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Output fromAm plifier Cycle 2 20 15 m V 10 5 0 1 5 9 13 17 21 25 29 33 37 41 45

Histg ramm ory em Cycle 2 3 2 1 0 1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46

No. of Ions

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Output from Am plifier Cycle 3 20 15 10 5 0 1 5 9 13 17 21 25 29 33 37 41 45 Histgram m ory em Cycle 3 No. of Ions 3 2 1 0 1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46

m V

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Output fromAm plifier Cycle 4 20 15 10 5 0 1 5

The ion which had about two times higher intensity was detected. It is counted only once (not twice) with TDC.

m V

9 13 17 21 25 29 33 37 41 45

Histgram m ory em Cycle 4 No. of Ions 3 2 1 0 1 5 9 13 17 21 25 29 33 37 41 45

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Output from Am plifier Cycle 5 20 15 10 5 0 m V

Two ions detected in succession!

17

25

45

13

21

29

33

37

41

Histgram m ory Cycle 5 em No. of Ions 3 2 1 29 45 1 5 9 13 17 21 25 33 37 41 0

The second ion can't be counted during dead time.

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Result of spectrum accumulation by TDC

m el sp od ectrum: No. of Ions 25 20 15 10 5 0

21

29

41

13

17

25

33

37

Histgram m ory Cycle 5 em No. of Ions 3 2 1 29 45 1 5 9 13 17 21 25 33 37 41 0

45

The ratio of the peak intensity isn't correct. A high intense peak shifts to low mass side.

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Continuous Averager

Continuous Averager
59us Timing Control Circuit High Voltage Pulser Amp ADC (8bit) Adder

Summing Memory

To Data System Intensity 15 28 .... No. of Data Points on a Spectrum (up to 256K points)

Simulation of spectrum accumulation by continuous averager

Output fromAm plifier Cycle 1 20 15 m V 10 5 0 1
50 40 30 20 10 0

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9 13 17 21 25 29 33 37 41 45
Cycle 1

mV

13

17

21

25

29

33

37

41

45

Simulation of spectrum accumulation by continuous averager

Output fromAm plifier Cycle 2 20 15 m V 10 5 0 1
50 40 30 20 10 0

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9 13 17 21 25 29 33 37 41 45
Cycle 2

m V

21

29

41

13

17

25

33

37

45

Simulation of spectrum accumulation by continuous averager

Output from Am plifier Cycle 3 20 15 10 5 0 1
50 40 30 20 10 0

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m V

9 13 17 21 25 29 33 37 41 45
Cycle 3

m V

17

25

45

13

21

29

33

37

41

Simulation of spectrum accumulation by continuous averager

Output from Am plifier Cycle 4 20 15 10 5 0 1
50 40 30 20 10 0

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m V

9 13 17 21 25 29 33 37 41 45
Cycle 4

m V

17

25

45

13

21

29

33

37

41

Simulation of spectrum accumulation by continuous averager

Output from Amplifier Cycle 5 20 15 10 5 0 1 5 9 13 17 21 25 29 33 37 41 45
Cycle 5 50 40 30 20 10 0 m V 17 25 45 1 5 9 13 21 29 33 37 41

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m V

Result of spectrum accumulation by continuous averager

No. of Ions

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The ratio of the peak intensity is correct. There is no shift of the ion peak.

m el sp od ectrum: 25 20 15 10 5 0

21

29

41

13

17

25

33

37

Cycle 5 50 40 30 20 10 0

m V

17

25

45

13

21

29

33

37

41

45

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Specifications
Mass resolution 6,000
FWHM, Reserpine m/z 609

Sensitivity Reserpine 10pg S/N>10

LC-ESI Flow rate: 0.2mL/min Mass chromatogram of m/z 609, RMS

Mass accuracy 5ppm RMS

With internal reference (Typically better than that!)

Only 3 analyzer parameters are critical Detection for routine DART analysis system
Ion Source Ion Transportation Analyser To the data collection system

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3 RP TMP1 RP

TMP2

1: Orifice 1 2: Peaks voltage 3. Multiplier V

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The 3 important parameters

1: Orifice 1: Typically 20V
Increase O1 to increase fragmentation

2: Peaks voltage (RF ion guide voltage)

Divide by 10 to estimate lowest detected m/z

3. Multiplier V: Typically 2200V to 2600V

Increase multiplier to increase signal (and noise)

Information from the TOF mass spectrum

Exact mass + isotope peaks: elemental composition Fragmentation: distinguish isomers Fingerprint pattern: material ID Ion abundance: quantitative analysis Other experiments: H/D exchange, derivatization, etc.

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Example: DART mass spectrum of a leaf

What is this?

100
80

304.154 290.174

Rel. Abund.

60 40 20 0

100

150

200

250

300

350

m/z

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We can treat this as an unknown

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Elemental compositions
Measured Exact Mass
Constraints

Candidate compositions
Isotope pattern matching

Ranked compositions
Elemental Composition Program

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We have a composition. Now what? m/z 304.1548 is C17H22NO4

Scopolamine
Fragments at m/z 138, 156 9 9
N

Cocaine
Fragments at m/z 182, 82
N O

138 C8H12 NO+

5 6 O O O H

O O

156 C8H14 NO2+

1 3

182 C10 H16 NO2+

2 4 2 8 o2 x y4 , l3 i - c (b a e 2 5 2 n zi do ) y- - m o c 8 l 2 6 2 xe y t- , hm y e l

2 0 4 e

2 1 4

2 2 4

2 0 2 2 1 2 2 2 2 2 3 2 ( m 2 3a 4 i )n 8 -l2i b 4 z 4a b 2 i [53 . 2y. 1 c o 2l c 6 t 4 a- 2 - n 2 ea7 4 b A c 4 ] o c r

2 7 2 t h y , l [ 1 eR

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API interface change potentials to LC Eluent fragmentation induce

Nebulizer Gas Desolvating Chamber

Desolvating Gas

Orifice2 Ion Guide Ring Lens Orifice1

Control fragmentation with Orifice 1 and Ring Lens potentials

RP

TMP

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Fragment spectrum increase cone voltage from 20 V to 60 V

Atropine 100
80

Scopolamine C8H12 NO+

138.089

290.174

C8H14 NO2+
156.099

Rel. Abund.

Scopolamine
304.154

60 40 20 0

C8H14 N+ 100 150 200 250 300

m/z

For comparison, m/z 305.1548 fragments from a dollar bill

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Cocaine
C10 H16 NO2+
100
80 Rel. Abund. 60 40 20 0
182.118

C17 H22 NO4+ C5H8N+

82.065

100

150

200

250

300

350

m/z

orwe can search for candidates from a list of target compounds.

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Components in a smokeless powder

SearchFromList Program

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Whew! Confused? Itll make more sense when you see it in the lab.