Spekroskopi Massa

(Part 1)
 Mass Spectrometry
A.) Introduction:
Mass Spectrometry (MS) measures the atomic or molecular weight of a ion from the
separation based on its mass to charge ratio (m/z)
- elemental composition of matter
- structures of inorganic, organic and biological molecules
- qualitative and quantitative composition of complex mixtures
- isotopic ratios of atoms in the sample

One of the MOST Routinely used Analytical Techniques

 Mass Spectrometry
Nobel Laureates:

Joseph John Thomson Francis William Aston

Physics 1906 Chemistry 1922
first mass spectrometer mass spectrometry of isotopes

John B. Fenn
Wolfgang Paul
Chemistry 2002
Physics 1989
electrospray ionization of
quadrupole and
biomolecules
quadrupole ion trap MS

Koichi Tanaka
Chemistry 2002
Matrix-assisted laser
Desoprtion/ionization (MALDI)

A Long and Continuing History of Achievements

 Mass Spectrometry • Qualitative Analysis
- Molecular Weight Determination
Mass Spectrometry Data - Structure Determination
• Quantitative Analysis
Quantitative Information

- Biotechnology
 analysis of proteins & peptides
Abundance

 analysis of oligonucleotides
- Pharmaceutical
 drug discovery, combinatorial chemistry
 pharmokinetics, drug metabolism
- Clinical
 neonatal screening, hemoglobin analysis
 drug testing
- Environmental
 water, food, air quality (PCDs etcs)
Qualitative Information - Geological
 oil composition
- Toxicology
- Forensics
 Mass Spectrometry
Advantages Over Atomic Optical
Spectrometric
• Detection limits three orders of
magnitude better
• Remarkably simple spectra that
are unique and easily interpreted
• Ability to measure isotopic ratios

Disadvantages
• Instrument costs are two to three
times higher
• Instrument drift that can be as
high as 5-10% per hour
• Interference effects
Atomic Weights in MS
• Discriminates among the masses of isotopes
- differs from other analytical techniques
• Atomic mass units (amu) or daltons (Da)
- Relative scale: 126 C  exactly 12 amu
- amu or Da equals 1/12 mass of 12 C
6
 1.66054x10 -24
g/atom
• All measured masses relative to 12 C
6
- 35
C has a mass 2.91407 C times 12
17 6
- C 12.0000 Da x 2.91407 = 34.9688 Da  atomic weight = 34.9688 g/mol
35
17
• Exact Mass (m)
- Sum of specific set of isotopes within compounds
 12
C1H4: m = 12.00000 x 1 + 1.007825 x 4 = 16.031 Da
 13
C1H4: m = 13.00335 x 1 + 1.007825 x 4 = 17.035 Da
 12
C1H32H1: m = 12.00000 x 1 + 1.007825 x 3 + 2.0140 x 1 = 17.037 Da
 3-4 significant figures to right of decimal
• Nominal Mass (m)
- Whole number precision in mass measurement
 12
C1H4: m = 12 x 1 + 1 x 4 = 16 Da
 13
C1H4: m = 13 x 1 + 1 x 4 = 17 Da
 12
C1H32H1: m = 12 x 1 + 1 x 3 + 2 x 1 = 17 Da
Atomic Weights in MS
• Chemical atomic weight or average atomic weight
n
A  A1 p1  A2 p2    An pn   An pn
i 1
 A1, A2, An :atomic masses in Da
 p1, p2, pn :fractional abundance of each isotope
 n :number of isotopes
• Weight of interest for most purposes
• Sum of chemical atomic weights for the atoms in the compound formula
- CH4 (m) =12.01115 + 4 x 1.00797 = 16.0434 Da
- typical atomic masses in periodic table

Boron: 10B 23% Zirconium: 90Zr 51.5% 91Zr 11.2%

11
B 100% 92
Zr 17.1 % 94Zr 17.4% 96Zr 2.8%
B (m) = (23x10 + 100x11)/123 Zr (m) = (51.5x90 + 11.2x91 + 17.1x92 + 2.8x96)/100
= 10.81 = 91.22
Molecular Formulas from Exact Molecular Weights
• Requires identification of molecular ion peak
• Exact mass needs to be determined
 High resolution instruments
 detect mass differences of a few thousands of amu
- Purine, C5H4N4 (m = 120.044)
- Benzamidine, C7H8N2 (m = 120.069)
- Ethyltolune, C9H12 (m = 120.096)
- Acetophenone, C8H8O (m = 120.058)
 Molecular ion peak is 120.070 ± 0.005  only C7H8N2 is possible formula
 Precision of a few parts per million is routinely possible
Molecular Formulas from Isotopic Ratios
• Low-resolution instrument  only differentiate whole
number masses
• Requires sufficiently intense molecular ion peak
• Requires accurate heights for (M+1)+ and (M+2)+

C6H4N2O4 C12H24

13
C 6 x 1.08 = 6.48
13
C 12 x 1.08 = 12.96%
2
H 4 x 0.0015 = 0.060%
2
H 24 x 0.0015 = 0.36%
15
N 2 x 0.37 = 0.74%

17
O 4 x 0.04 = 0.16% (M+1)+/M+ = 13.32%

(M+1)+/M+ = 7.44%
Resolution in MS (R)
• differentiate between masses
R = m/Dm
or
( m1  m2 )
R 2
m

• Higher the number the better the resolution

 500,000 is better than 500
 Resolution of 4000 would resolve peaks occurring at m/z values of:
 400.0 and 400.1
 40.00 and 40.01
Identification of odd electron ions – The Nitrogen Rule
• Based on an anomaly in the relationship between the atomic weights and
valences of the common elements
• An organic molecule containing the elements C, H, O, S, P or halogen
- odd nominal mass if it contains an odd number of nitrogen atoms
- even nominal mass if it contains an even number of nitrogen atoms
(including 0)

Elemen Compound MW
At Wt Valence
t
CO2 44
H 1 1
CO 28
C 12 4
CH4 16
N 14 3
O 16 2 HCO2H 46
F 19 1 HCl 36/38
S 32 2 HCN 27
Cl 35/37 1 N2 28
NH3 17
Even nominal mass  even valence
Odd nominal mass  odd valence

Nitrogen is the exception

Double Bond Equivalent (D)
• number of rings or double bonds that an ion contains
• Calculated from the elemental formula as follows:

where:
 D is unsaturation
 imax is the total number of different elements
 Ni the number of atoms of element i and
 Vi the valence of atom i
• valence of 1 (H, F, Cl), valence of 2 (O, S).
• valence of 3 (N, P), valence of 4 (C, Si).

Hexane: C6H14 Hexene: C6H12

Mass of molecular ion: 86 Mass of molecular ion: 84
D = 1+1/2((6x(4-2) + 14x(1-2)) = 0 D = 1+1/2((6x(4-2) + 12x(1-2)) = 1
Basic MS Instrument Design
• Atomic Mass Spectrometry involves the following steps:
- atomization
- conversion of atoms to ions
 most ions are single charge

- separation of ions based on mass-to-charge ratio (m/z)

- Counting the number of ions of each type or measuring the ion current
• Principal Components
- Vacuum system  maintain low pressure(10-5 to 10-8 torr)
- Inlet: introduce m-amount of sample into ion source
- Ion source: sample converted into gaseous ion by bombardment with:
 Electrons

 Photons

 Ions

 Molecules

 Thermal/electric energy

- Positive/negative ions accelerated

into mass analyzer
- Mass analyzer: disperse ions (m/z)
- Transducer: convert beam of ions to
electrical signal
Types of Atomic and Molecular MS
• Thermal ionization & Spark source  first MS
• Inductively coupled plasma (ICP)  current common approach
- Differ by types ion sources and mass analyzer
MS Theory:

1.) Mass analyzers use electric and magnetic fields to apply a force on charged
particles
F = ma (Newton's second law)

F = e(E+ v x B) (Lorentz force law)

where:
F - force applied to the ion m - mass of the ion
a - acceleration e - ionic charge
v x B - vector cross product E - electric field
of the ion velocity and the applied
magnetic field

2.) Force is therefore dependent on both mass and charge

- spectrometers separate ions according to their mass-to-charge ratio
(m/z)

- not by mass alone

Inlet Systems:

• Introduce sample into ion source with minimum loss of vacuum

• Spectrometer equipped with multiple inlets for different sample types
 Batch Inlet
 Direct Probe Inlet
 Gas Chromatography
 Liquid Chromatography
Inlet Systems:
External (Batch) Inlet Systems:
• Simplest
• Gas & liquid samples (bp < 500oC)
• Sample heated (<400 °C) in small external oven
• Sample pressure 10-4 to 10-5 torr
• Vapor admitted to ionizer through valve
• Gas stream added to analyte
Inlet Systems: Direct probe inlet system for solids

The direct probe is only ¼" in diameter.

Probe moves through various lock system stages permits
for a step-wise increase in the vacuum

Direct Probe
• Solids and non-volatile samples
- Less sample is required & wasted (few ng)
- sample held on the surface of glass or aluminum capillary tube, fine wire or small cup

• Sample vial inserted through air-lock into ionizer chamber

- Lock system minimizes amount of air that must be pumped from system

• Vial heated to vaporize sample

• Vial can be reduced to capillary or surface plate for small quantities

Inlet Systems:
• LC & GC coupled to mass spectrometer

• Permits separation and determination of components for complex mixtures

 Requires specialized inlet systems
 Major interface problem – carrier gas dilution
 Jet separator (separates analyte from carrier gas)
- Lighter carrier gas deflected by volume
- Heavier sample travels in straight line