Mass Spectrometry
Simplest form of mass spectrometer performs 4 essential functions: (under vacuum 10-6 mm Hg)
It subjects vaporized molecules to bombardment by a stream of high-energy electrons, converting these molecules to ions These ions are then accelerated in an electric field The accelerated ions are then separated according to their mass-to-charge ratio in a magnetic or electric field The ions that have a particular mass-to-charge ratio are then detected by a device that counts the number of ions striking it
MS uses magnetic and electric fields to exert forces on charged particles (ions) in a vacuum. Therefore, a compound must be charged or ionized to be analyzed by a mass spectrometer.
Mass Spectrometry
Atomic and molecular weight are expressed in Atomic Mass Unit (amu). The amu is based on a relative scale in which
12C
Each isotope has specific exact mass. MS is interested in exact mass up to 3-4 figures after decimal point (high resolution HR). Chemical atomic weight is the average weight taking into account the mass of each isotope and their natural abundance.
Vacuum System
Signal processor
m/z
MS : Ionization
1. Gas Phase:
1. Electron ionization (EI) 2. Chemical ionization (CI) 3. Field Ionization (FI)
2. Desorption
1. 2. 3. 4. 5. 6. 7. 8. Field Desorption (FD) Fast Atom Bombardment (FAB) Secondary Ion Mass Spectrometry (SIMS) Laser Desorption (LD) Plasma Desorption (PD) Thermal Desorption Thermospray ionization (TS) Electrospray (ES)
Mass Spectrometer
If a molecule absorb an electron (creating a Negative Ion), It will be absorbed by the repeller plate The repeller plate (Positively charged) directs the ions through a series of accelerating plates Kinetic Energy : 1 mv2 = eV 2
m = H2 r2 e 2v
1. 2. 3. 4. 5. 6.
Magnetic sector Electrostatic and Magnetic sector Quadrupole MS filter Ion trap analyzers TOF Time-of-Flight FT-MS
Vacuum System
Amplifying signal
Continuous Dynode electron multiplier Provide high gain and nanosecond response time.
Mass Spectrum
Ionization: CH3OH + eCH3OH+ + 2em/z 32 Molecular ion Fragmentation: [CH3OH]+ [CH2OH] + + H
m/z 31
CH3+ + HO m/z 15 B Base Peak 100%
CHO+ + H2
m/z 29
B+
Benzamide: EI
M+
77
44 - NH2
C O
NH2
C6H5+ m/z 77
105
3-methyl-6-i-PropylCyclohex-2-ene-1-one
MW = 152
12.011
12.0000 (std) 98.89 13.00336 1.11 14.0031 15.0001 15.9949 16.9991 17.9992 99.64 0.36 99.76 0.04 0.20
14.0067
15.9994
Mass 18.9984 27.9769 28.9765 29.9738 31.9721 32.9715 33.9679 34.9689 36.9659
% Abundance 100.0 92.17 4.71 3.12 99.64 0.76 4.20 75.77 24.23
32.066
35.4527
126I
126.9045
126.9045
100.00
Atomic weight Br: [% 79Br x 78.9183] + [% 81Br x 80.9163] [50.52 x 78.9183] + [49.48 x 80.9163] = 79.9094
Molecular formula
e.g C7H7NO :
M+ (molecular ion) m/z M mass of the most abundant isotope 7 x 12C = 84 7 x 1H = 7 1 x 14N = 14 1 x 16O = 16 m/z 121
%(M+1) = (1.1 * #C) + (.38 * #N) = 8.08 % %(M+2) = (1.1 * #C)2/200 + (.2 * #O) = 0.5% For C, H, N,O, F, P composition
Even Mass : From even mass M+ comes from rearrangement or 2 bond breaking Odd Mass : From even mass M+ comes from single bond breaking
even Mass : from Odd Mass M+ comes from single bond breaking
C11 H20 CH4 + O = C10 H16 O C10 H16 O CH4 + O = C9 H12 O2 C9 H12 O2 CH4 + O = C8 H8 O3
=> =>
CH2 CH7
1H
28Si
31P 32S 35Cl 79Br 127I
=>
=> => => => =>
C2H4
C2H7 C2H8 C2H11 C6H7 C10H7
12
=>
Nitrogen rule
Molecular ion => 27 odd! 27 / 13 = # carbons = 2 Basic formula C2 H3 One Nitrogen C2 H3 - CH2 + N
H C N
Basic formula C2 H2
H C C H
Nitrogen rule
OH NH2
Isotope peaks
Usually [M+2] peak is very small
Except for:
M Sulfur :
32S
M+1
M+2
34S
: 100
: 4.4
Silicon :
Chlorine : Bromine :
28Si:
100
29Si:
5.2
30Si
: 3.35
: 32.5 : 98
35Cl
: 100 : 100
37Cl
79Br
81Br
M-H
[12CH279Br]+
[13CH379Br]+
[12CH281Br]+ [13CH381Br]+
Bromine
79Br 81Br
79Br 81Br
and 81Br
Chlorine
35Cl
2 Cl
P (2 35Cl) = (0.75 )2 = 0.563 P (35Cl 37Cl) + P (37Cl 35Cl ) = (0.75 ) (0.25 ) + (0.25 ) (0.75 ) = 0.375 P (2 37Cl) = (0.25 )2 = 0.063 [M+] / [M + 2] / [M + 4] = 100 / 66 / 11
37Cl
=> .25
81Br
79Br
=> 49%
37Cl 81Br
At very high resolution, the 2 (M + 2) peaks can be distinguished (separated by 0.001 Dalton)
In percentage: n x 1.1 %
Generally speaking, The size of the [M+1]+ ion can be used to figure out the number of carbons in a molecule
For example, if molecular ion is observed at m/z 118
When there are many carbons, isotopic pattern for 13C adds up to chlorine pattern
Solving problems in MS
Try to identify the Molecular Ion or decide if it is present (most critical step in solving a structure)
Check if [M+1]+ ion is too large to accommodate reasonable number of carbons. (the [M+1]+ ion might be the very small M+ instead!) Determine the first loss from proposed molecular ion. Some loss are impossible (e.g 12, 14, 23 daltons) Does the spectrum appear dirty? (lots of small peaks even at high mass) If GC of the comopund Is available, compare retention time
Examine ion cluster for isotopic natural abundance (look for special heteroatom pattern). Try to calculate number of carbons
Solving problems in MS
From the overall appearance: is it a fragile compound? Is it likely to be aromatic or aliphatic? Look in the low mass ions. Do you see any clues of the family of compounds that you might be dealing with? Make a list of suggested losses from the molecular ion and try to make a pattellsrn from them. Look for intense odd-electron ions in the spectrum: this is almost impossible in compounds containing Nitrogen! These provides clues for rearrangements (retro Diels Alder, McLafferty) Speculate on the structure using all that information Index of Hydrogen deficiency
CxHyNzOn
Index = x y + z +1
M-31 CH3O
M-32 CH3OH M-35 Cl
M-36 HCl
M-42 CH2=C=O, CH2=CH-CH3 M-43 CH3CO , C3H7 M-44 CO2 M-45 CH3CH2O , CO2H
M-27 HCN
M-28 H2C=CH2 , CO M-29 CH3CH2 , HCO
Resolution
Mass Defects
By definition the atomic weight of 12C is 12.0000 Daltons All other elements are determined relative to 12C Their difference to nearest integral is called Mass Defect Light element has small positive mass defect Vast majority of elements have substantial negative mass defect
Mass Defects
The Mass Defect of a C23H35O2Si is shown here on a low resolution MS (+/- 0.1)
Mass Defects
Mass defect brings the maximum of the C23H35O2Si peak not at 371 but at 371.25 If the Mass spectrometer measure the mass with a resolution of 1, the sensitivity is reduced.
Greatest application of Mass Defect is surely very high resolution, which provide mass with .001 to .0001 accuracy
High resolution:
Exact Mass can provide Molecular Formula With sufficient accuracy, unique molecular formula can be determine
e.g. Distinguish CO, N2, CH2N and C2H4 (all having m/z 28)
CO
12C
16O
N2
14N 14N
CH2N
12C
C2H4
12C
C14H8N2O3Cl
Mass Defect of C14H8N2O3Cl: (8 x .0078) + (2 x .0031) + (3 x -.0051 ) + (-.0311 ) = .0202 With sufficiently high resolution MS, it is possible to propose a unique empirical formula for an ion.
Nitrogen rule M+ even even # of N 0, 2, 4, M+ odd odd # of N 1, 3, 5, Index of Hydrogen deficiency Index = C H/2 X/2 + N/2 + 1 e.g. C7H7NO Index = 7 - 3.5 + 1 = 5 Hydrogen deficiency can be unsaturation (multiple bonds) or cyclic structure R-CN
I=2
I=1 (1 DB + cycle) I=4 (3 DB + cycle)
Index of hydrogen deficiency : General Index = IV (I/2) + (III/2) +1 IV any tetravalent atom (e.g. C, Si..)
+ CH3-S-CH3
OI=0
+ CH3-N
I=1
O O-
CH3 + CH -P-CH
3
phosphine
OI=0
Ionization Techniques
Electron Impact (EI) Under these conditions, very energetic Ions are produced => Fragmentation M.+ is often weak e.g. ROH often loose H2O
2- SIMS : other source of energy is a beam of ions (Secondary Ion Mass Spectroscopy): Typically Argon (FAB)
Field Desorption methods give a high concentration of Molecular ion => Particularly useful for identification of unknown
The target is bombarded with a fast atom beam (for example, 6 keV xenon atoms) that desorb molecular-like ions and fragments from the analyte.
Xe
e-
Xe+ + Xe
Xe + Xe+
Xe
Cluster ions from the liquid matrix are also desorbed and produce a chemical background that varies with the matrix used.
FAB: Benefits
Sample introduction can be through direct insertion probe or LC/MS (continuous-flow FAB).
Benefits
rapid, simple relatively tolerant of variations in sampling
FAB: Limitations
Limitations high chemical background defines detection limits may be difficult to distinguish low-molecular-weight compounds from chemical background analyte must be soluble in the liquid matrix no good for multiply charged compounds with more than 2 charges requires a high concentration of the organic liquid matrix (typically 80 to 95% glycerol) which limits sensitivity Mass range Moderate Typically ~300 Da to about 6000 Da.
3. Time of Flight
secondary ions or
laser generated photons
These pulses have typically a frequency of 10-50 kHz and lifetime 0.25ms The ions produced are then accelerated by electric field pulse (103-104 V) that has the same frequency as ionization pulse but lags behind. The accelerated ions then pass into a field-free drift tube (about 1 m long)
Tandem-MS
Time of Flight
105
MW = 164
Organometallic compounds in MS
It is often possible to determine molecular weight of a compound by MS
Molecular ion is usually present with Laser desorption. However, ions produced That way comes from condensed phase (solid/liquid) and structure in these phase Might be very different from the one in gaz phase. With FAB most intense cation peak is the protonation ion (M+1) peak. Anion (M-1) can also be formed.
=> 51%
81Br
0.37 x 0.51 = 19% 0.37 x 0.49 = 18.1% 0.63 x 0.51 = 32.1% 0.63 x 0.49 = 30.9%
50.2%
268 m/z
Metastable ions
Some ions have so short lifetimes that they dissociate while moving through the spectrometer.
An ion of mass m1 is accelerated after initial ionization But a different ion m2 (daughter ion) passes the magnetic analyzer The resulting peak is neither m1 or m2 but appear at m* (metastable) m* = (m2)2/m1
These metastable ions are formed during 10-5 s (time spent between electrostatic and magnetic analyzer) are quite broad but provide direct information about ion reactions
Platinum
Tungsten
Index
MS-fragmentation