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time or volume
Mobile Phase
gas (GC) water (LC) organic solvent (LC) supercritical fluid (SCFC)
Types of Chromatography
Classification by mobile phase:
Gas - Gas chromatography (GC)
1951 Martin and James (fatty acids)
Gas - solid
Gas - liquid
Instrumentation for GC
Carrier gas
N2, He, H2
oven
Modes of GC Separation
Capillary (open tubular)
Inner wall modified with thin (1 m) film of liquid 0.3 - 0.5 mm ID; 10 - 50 m length
Packed
Solid particles either porous or non-porous coated with thin (1 m) film of liquid 1 - 8 mm ID; 1 - 10 m length
GC Liquid Phase
Low volatility High bp Chemically unreactive Examples:
1-squalene Tetrahydroxyethylenediamine Carbowax (polyethylene glycol)
GC - Modes of Separation
Isothermal (GC) Programmed temperature (GC)
Raising column temperature (GC)
Decreases retention time Sharpens peaks
Rapid response Linearity - concentration range over which signal proportional to concentration Stability with respect to noise (baseline noise) and time (drift)
Detectors for GC
MS Components
Ionization source
Analyzer
Detector
Ionization Methods
Electron capture (EC)
70 eV e- neutral molecule energetic molecular ion hard; fragmentation
Ionization Methods
Electrospray (ESI)
generation of ions by desolvation or desorption of charged liquid droplets
Types of MS Analyzers
Quadrupole - most common
Ion trap
Time of Flight (TOF)
tr
Retention Time
Mass Spectrum
X - axis: m/z mass - based on 12C 12.0000 Y - axis: relative abundance
usually normalized wrt largest line (base peak) 0 - 100 %
Molecular Ion
Ion whose mass equals that calculated from the molecular formula using the masses for each element which have the highest natural abundance; often tallest peak in highest m/z group Base peak - most intense peak in spectrum; not necessarily the molecular ion peak!
m/z 12 13 14 15 16 17 28 29 30 31 32 33 34
Rel. Abundance 0.33 0.72 2.4 13. 0.21 1.0 6.3 64 3.8 100. 66. 0.98 0.14
Rel. Abundance, %
15
10
13
16
19
22
25
28
31
34
37
m/z
40
Figure taken from Rubinson, K.A. Chemical Analysis Boston: Little, Brown, 1987.
Lessons:
Single charge most common so m/z usually equates to mass (EI/CI) Ions may fragment
EX: CH3OH loses H+ readily Observe: CH3O +
Isotopes
Most abundant isotope of an element is set to 100% Abundance of other isotopes are normalized with respect to it
3 Classes of Isotopes
A - only a single isotope
EX: F, P, I
A+1 - two isotopes with significant relative abundance differing by 1 mass unit
EX: H, C, N
A+2 - two isotopes with significant relative abundance differing by 2 mass units
EX: Cl, O, S
12
100
13
1.08
A+1
N 14 100 15 0.36
Cl
35
100
37
32.5
A+2
Cl2 Revisited
Two isotopes: 35Cl and 37Cl Three possible species formed: and 37Cl37Cl Relative abundance:
35Cl35Cl:
35Cl35Cl, 37Cl35Cl,
1.0 x 1.0 = 1.0 37Cl35Cl and 35Cl37Cl: 1.0 x 0.325 = 0.325 each or 0.66 37Cl37Cl: 0.325 x 0.325 = 0.106
So, answer: 3 peaks at 70, 72, and 74 with relative intensities of 100, 32.5, and 10.6 %
32
100
33
0.80
34
4.40
A+2
Problem 1:
m/z Rel. Abundance m/z Rel. Abundance 35 36 43 47 48 49 50 51 1.8 1.3 2.1 13.7 5.8 100 3.5 32 57 83 84 85 86 87 88 1.5 2.1 64.2 2.3 42.8 0.8 7.1
A +2 A +2 A +2