Trinity College Dublin School of Chemistry 16 Dec 2009

WelcomeTo the

Mass Spectrometry Unit

Dr. Martin Feeney Room 0.5 Dr. J. Bernard Jean-Denis Chemistry building

Trinity College Dublin School of Chemistry p 1

1. Introduction to mass spectrometry 2

2. Sample Preparation 7 a. Choice of Solvents b. Sample Solubility c. Sample Concentration d. Sample Vials e. Things to avoid in Sample Prep.

3. Sample submission form 9 a. Sample ID b. Proposed Structure c. Sample Handling/Storage

4. Results 12

5. MS queries 13

Table of Contents

Trinity College Dublin School of Chemistry p 2

Introduction

1. Introduction to mass spectrometry

a- ESI- TOF-MS

b- GC- TOF-MS

c- LDI-Q-TOF-MS

d- ESI-LIT-MS 2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

There are a number of instruments in the mass spectrometry facility of the School of chemistry that can offer a wide range of techniques and experiment types, such as MS/MS experiments or mass accuracy determination

MS/MS experiments offers structural information of the moleculeTOF, Time-of-flightESI, Electrospray ionisationMALDI, Matrice assisted laser desorption ionisationLIT, Linear ion trap

Techniques Instrumentations Manufacturer Mass accuracy

MS/MS

LC-ESI-TOF- MS LCT Classic Waters Yes No

GC-TOF-MS GCT Premier Waters Yes No

MALDI-QTOF-MS Maldi-Qtof Premier Waters Yes Yes

LC-ESI-LIT-MS API 2000 Applied Biosystems

No Yes

Introduction

Principle

An electric field sufficiently intense to disperse analyte solution emerging from the spray needle into a fine spray required a potential difference of up to sereval kV between that tip and the orifice leading into the vacuum system.The ions will be separate according to their mass-to-charge ratio through a TOF mass analyser before the determination of the accurate mass.

1. Introduction to mass spectrometry

a- ESI- TOF-MS

b- GC- TOF-MS

c- LDI-Q-TOF-MS

d- ESI-LIT-MS 2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Trinity College Dublin School of Chemistry p 3

Sample characteristics

Charged or partially charged polar compounds.Sample might need ionisation agent, such as NaCl, Ammonium acetate,...

Solvent choice -> see page 7

ESI- TOF-MS

ElectroSpray Ionisation coupled toTime-Of-Flight mass spectrometer

References:

Dole M et al., 1968. J. Chem. Phys., 49 (5), p2240.Fenn JB et al., 1985. Anal. Chem., 57(3), pp675-679Wiley WC & McLaren IH, 1955. Rev. Scientific Instruments, 26, (12), pp1150-1157.

LCT Classic, Waters

Introduction

1. Introduction to mass spectrometry

a- ESI- TOF-MS

b- GC- TOF-MS

c- LDI-Q-TOF-MS

d- ESI-LIT-MS 2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Trinity College Dublin School of Chemistry p 4

Principle

The sample coming from GC effluent could be ionised by three ionisation methods, which are EI (Electron Impact), CI (chemical Ionisation) and FD (Field desorption).The ions will be separate according to their mass-to-charge ratio through a TOF mass analyser before the determination of the accurate mass.

Sample characteristics

Sample must be thermally volatile and stableLow mass range (typically lower than m/z 650)

Solvent choice -> see page 7

GC- TOF-MS

Gas Chromatography coupled toTime-Of-Flight mass spectrometer

References:

Lampman GM et al., 2010. Spectroscopy, International Edition, 4thedition.Wiley WC & McLaren IH, 1955. Rev. Scientific Instruments, 26, (12), pp1150-1157.

GCT Premier, Waters

Introduction

1. Introduction to mass spectrometry

a- ESI- TOF-MS

b- GC- TOF-MS

c- LDI-Q-TOF-MS

d- ESI-LIT-MS 2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Trinity College Dublin School of Chemistry p 5

Principle

The sample will be mix with a matrix*. Then, A laser will desorb the mixture and generate ions with high kinetic energy. The ions will be separate according to their mass-to-charge ratio through a TOF mass analyser before the determination of the accurate mass.

MS/MS experiments can be done.Possibility of using ESI.

* DCTB, CHCA, DIT, DHB.

Sample characteristics

Generally speaking, samples which could not be ionised by ESI and non volatile

Solvent choice -> see page 7

LDI- TOF-MS

Laser Desorption Ionisation coupled toTime-Of-Flight mass spectrometer

References:

Lampman GM et al., 2010. Spectroscopy, International Edition, 4thedition.Wiley WC & McLaren IH, 1955. Rev. Scientific Instruments, 26, (12), pp1150-1157.

MALDI Q-TOF, Waters

Trinity College Dublin School of Chemistry p 6

Introduction

1. Introduction to mass spectrometry

a- ESI- TOF-MS

b- GC- TOF-MS

c- LDI-Q-TOF-MS

d- ESI-LIT-MS 2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Principle

Quadrupole mass analysing device electric field is used to separate ions according to their mass-to-charge ratio as they pass along the central axis of four parallel equidistant metal rods that have fixed (DC) and alternating (RF) voltages.

MSn , multiple MS/MS experiments can be done.

Sample characteristics

Charged or partially charged polar compounds.Sample might need ionisation agent, such as NaCl, Ammonium acetate,...

Solvent choice -> see page 7

ESI- LIT-MS

Electrospray ionisation coupled to a linear iontrap mass spectrometer

References:

Lampman GM et al., 2010. Spectroscopy, International Edition, 4thedition.Paul W, Reinhard HP & VonZahn U, 1958., Z. Phys., 152 (2), p143-182.

API 2000, Applied Biosystems

Sample Preparation

1. Introduction to mass spectrometry

2. Sample preparation

a- Choice of solvent Sample solubility

b- Sample concentration

Sample vials

3. Sample submission form

4. Results

5. MS queries

Choice of solvent

MALDI-QTOF

GC-TOF-MS

ESI-TOF

MeOH, ACN, DCM, CHCl3,

EtOAc

MeOH, ACN, DCM, CHCl3,

EtOAcMeOH, ACN,

H2O

H2O, DMSO

Sample solubility

Sample must be fully dissolved.

In the event of partial solubility, filter or centrifuge the sample. Then, decant the supernatant to a clean vial for MS analysis.

Trinity College Dublin School of Chemistry p 7

Trinity College Dublin School of Chemistry p 8

Sample Preparation

1. Introduction to mass spectrometry

2. Sample preparation

a- Choice of solvent Sample solubility

b- Sample concentration

Sample vials

3. Sample submission form

4. Results

5. MS queries

Sample concentration

Mass Spectrometry is at least 20 times more sensitive than NMR.

Do not submit the sample for MS straight from the NMR tube, please.

MALDI-QTOF / ESI-TOF GC-TOF-MS

~ 1mg.mL-1 ~ 5mg.mL-1

Sample Vials

Use 2 mL HPLC vials with dimension (12 x 32 mm (diameter x height)). Add anything between 0.5 to 1.5 mL solvent but please do not fill to the top of the

vial.

Things to Avoid in Sample Preparation: - salt buffers such as phosphates, citrates, borates, etc. - detergents - inorganic acids (H2SO4, H2PO4, etc)

For HRMS, your sample must be pure.

Trinity College Dublin School of Chemistry p 9

Sample submission form

Fill-in where there is a star

Draw the proposed structure, (see example in red)

O

Exact Mass: 140.1201Formula : C9H16O

Do not forget to indicate the solvent of dilution

X

XXX 001

YYYY-MM-DD

XXX@tcd.ie

XXX XXXZZZ ZZZ

X

X

1. Introduction to mass spectrometry

2. Sample preparation 3. Sample submission form

a- One sample

b- More than one sample 4. Results

5. MS queries

One sample

Trinity College Dublin School of Chemistry p 10

Sample submission form

1. Introduction to mass spectrometry

2. Sample preparation 3. Sample submission form

a- One sample

b- More than one sample 4. Results

5. MS queries

See Attachment

X

4 samplesYYYY-MM-DD

XXX@tcd.ie

XXX XXXZZZ ZZZ

X

X

More than one sample

O

O O

Exact Mass: 140.1201Formula : C9H16O

Exact Mass: 140.1201Formula : C9H16O

Exact Mass: 140.1565Formula : C10H10

Exact Mass: 142.1358Formula : C9H16O

XXX 001 XXX 002

XXX 004XXX 003

Trinity College Dublin School of Chemistry p 11

Results

1. Introduction to mass spectrometry

2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Experimental Mass

Theoritical Mass

O

Δm *

Mass Accuracy **

Unsaturationdegree

IsotopicPattern

matching

Empirical formula

*, |Expected mass – Theoretical mass|

* *, [Δm/(integer value of m)]x106

Name (Supervisor), XXX 001

Trinity College Dublin School of Chemistry p 12

MS queries

1. Introduction to mass spectrometry

2. Sample preparation 3. Sample submission form

4. Results

5. MS queries

Any questions regarding the mass spectrometry service in TCD then please ask Dr. Martin Feeney or Dr. Bernard Jean-Denis.

The mass spectrometry main lab phone number is 01-896 2768.

If you would like to attend the MS course taught in the Dublin Chemistry programme, please contact Dr. Jimmy Muldoon.

The course is usually scheduled for January-March of every year. Alternatively check the website:

http://www.dublinchemistry.ie