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ANÁLISIS QUÍMICO III

SECCIÓN ESPECTROMETRÍA DE MASAS

INSTRUCTOR: MARIANNY Y COMBARIZA

UNIVERSIDAD INDUSTRIAL DE SANTANDER

MMXX

INTRODUCTION TO MS

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Scientists Shrink Chemistry Lab to Seek Evidence of Life on Mars

https://www.nasa.gov/feature/goddard/2018/moma
https://www.youtube.com/watch?time_continue=90&v=ZSAzWBz6rNY&feature=emb_logo

Mass spectrometry
Foundations of mass spectrometry: end of the nineteenth century.

The first analyses of small molecules were reported in 1912 by

Joseph J. Thompson.

• “I have described at some length the application of Positive

Rays to chemical analysis; one of the main reasons for writing
this book was the hope that it might induce others, and
especially chemists, to try this method of analysis. I feel sure
that there are many problems in chemistry, which could be
solved with far greater ease by this than any other method. The
method is surprisingly sensitive -- more so than even that of
spectrum analysis, requires an infinitesimal amount of material,
and does not require this to be specially purified; the technique
is not difficult if appliances for producing high vacua are
available.”

J. J. Thomson, October 4, 1913

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Thomson´s cathode ray tube

The Nobel Prize in Physics 1906 was awarded to Joseph John Thomson "in recognition of the great
merits of his theoretical and experimental investigations on the conduction of electricity by gases.”

The Nobel Prize in Physics 1906. NobelPrize.org. Nobel Media AB 2020. Sat. 16 May 2020. <https://www.nobelprize.org/prizes/physics/1906/summary/>
https://edu.rsc.org/feature/mass-spectrometry-the-early-days/2020189.article

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Available via license: CC BY-NC 4.0

https://www.lindau-nobel.org/blog-how-to-weigh-an-atom-francis-w-astons-mass-spectrograph/

The Nobel Prize in Chemistry 1922 was awarded to Francis

William Aston "for his discovery, by means of his mass
spectrograph, of isotopes, in a large number of non-radioactive
elements, and for his enunciation of the whole-number rule.”
The Nobel Prize in Chemistry 1922. NobelPrize.org. Nobel Media AB 2020. Sat. 16 May 2020.
<https://www.nobelprize.org/prizes/chemistry/1922/summary/>

Work
Harold Clayton Urey "for his
discovery of heavy
hydrogen."
Harold C. Urey – Facts. NobelPrize.org.
Nobel Media AB 2020. Sat. 16 May 2020.

Work
Prof Robert F. Curl, Jr., Rice University, Houston, USA,
Prof Sir Harold W. Kroto, University of Sussex,
Brighton, U.K., and Prof Richard E. Smalley, Rice
University, Houston, USA, for their discovery of
fullerenes (carbon footballs).

Press release. NobelPrize.org. Nobel Media AB 2020. Sat. 16 May 2020. <https://www.nobelprize.org/prizes/chemistry/1996/press-release/

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1970s, transferring macromolecules to ions in the gas phase,

termed desorption technology. Basis for the revolution in this
field during the past twenty years.

Two principles for causing

proteins to transform into the
gas phase without losing
their structure and form.

John B. Fenn. Sample is sprayed using a strong electrical field

to produce small, charged, freely hovering ions.

Koichi Tanaka. Intense laser pulse. Molecules take up some of

the energy of the laser pulse and become released as free
ions.

M I L E S TO N E S T I M E L I N E
1910 The beginnings (Milestone 1)

1929 Development of ionization methods (Milestone 2)

1939 Environmental analysis (Milestone 3)

1946 Time of flight (Milestone 4)

1949 Trapping mass analyzers (Milestone 5)

1953 Quadrupole and triple-stage quadrupole mass filters (Milestone 6)

1956 Small-molecule analysis (Milestone 7)

1959 Separations (Milestone 8)

1959 Peptide sequencing (Milestone 9)

1961 Inductively coupled plasma mass spectrometry (Milestone 10)

1962 Imaging mass spectrometry (Milestone 11) https://www.nature.com/milestones/milemassspec/pdf/milemassspec_all.pdf

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1963 Carbohydrate analysis (Milestone 12)

1966 Tandem mass spectrometry (Milestone 13)

1966 Metabolomics (Milestone 14)

1968 Electrospray ionization (Milestone 15)

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1975 Medical applications (Milestone 16)
 1953 Quadrupole and triple-stage quadrupole mass filters (Milestone 6)

1956 Small-molecule analysis (Milestone 7) 1/07/20

1959 Separations (Milestone 8)

1959 Peptide sequencing (Milestone 9)

1961 Inductively coupled plasma mass spectrometry (Milestone 10)

1962 Imaging mass spectrometry (Milestone 11)

1963 Carbohydrate analysis (Milestone 12)

1966 Tandem mass spectrometry (Milestone 13)

1966 Metabolomics (Milestone 14)

1968 Electrospray ionization (Milestone 15)

1975 Medical applications (Milestone 16)

1978 Selected reaction monitoring (Milestone 17)

1985 Matrix-assisted laser desorption/ionization (Milestone 18)

1991 Structural biology applications (Milestone 19)

1993 Proteomics (Milestone 20)

1995 Post-translational modification analysis (Milestone 21)

1999 Interactome analysis (Milestone 22)

https://www.nature.com/milestones/milemassspec/pdf/milemassspec_all.pdf
4 | O CTOBER 2015 www.nature.com/milestones/mass-spec

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WHAT IS MASS SPECTROMETRY?

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Zeroth law of mass spectrometry

“Never make the mistake of calling it ‘mass

spectroscopy’. Spectroscopy involves the

absorption of electromagnetic radiation, and

mass spectrometry is different. The mass

spectrometrists sometimes get upset if you

confuse this issue”

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WHAT IS MASS SPECTROMETRY?

WHAT? HOW?
Generate ions from organic or
inorganic compounds. Single Thermally. Electric fields.
ionized atoms, clusters, Impact (electrons, ions,
molecules, fragments or photons, energetic neutral
associates. atoms, heavy cluster ions).

Separate ions according to Static or dynamic E or B.

mass-to-charge ratio (m/z) Time (field free regions).
and detect ions quali- and
Electron multipliers/
quantitatively by their m/z and
microchannels
abundance

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MASS SPECTROMETER

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MASS SPECTRUM
2D Representation of ION ABUNDANCE vs m/z

m= mass number
z= elementary charge
Rel. Intensity

If z=1 then m/z ~ m

Th (Thomson, not SI)

Da (Daltons, not SI)
u (unified atomic mass)
https://www.bruker.com/fileadmin/user_upload/8-PDF-Docs/Separations_MassSpectrometry/Literature/literature/ApplicationNotes/MT-112_Intact_Protein_2012_6-seitig_eBook.pdf

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M+.= Molecular Ion

Base peak = Most abundant
.
Fragment ions = m/z 12-15 loss of H and H2
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BASIC MS TERMINOLOGY
•Background signal–The signal output when a blank is measured.

•Noise (N)– Fluctuation in the instrument background signal (𝛔 of background

signal)

•Signal-to-noise ratio (S/N)–The ratio of the analyte signal to the noise measured
on a blank.
S/N > 10
GOOD SIGNAL ON A MS: S/N > 104
S/N > 1000 (invisible noise)

•Sensitivity–The signal response of analyte normalized to the amount of analyte

giving rise to the response. Slope of the calibration curve.

SEE PAPERS ON MOODLE!

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m/z Theor ratio

92 100
93 7.7
94 0.3

m/z 92 ∼ 13 times m/z 93 ∴ S/N m/z 92 ∼ 13 times S/N m/z 93

m/z 92 ∼ 333 times m/z 94 ∴ S/N m/z 92 ∼ 333 times S/N m/z 94

m/z 93 ∼ 25.6 times m/z 94 ∴ S/N m/z 93 ∼ 25.6 times S/N m/z 94
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BASIC MS TERMINOLOGY

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GAS PHASE ION CHEMISTRY

CHEMISTRY LABORATORY TO
MASS SPECTROMETER
STUDY IONS IN THE GAS PHASE

Creation of Mass Spectra

Fragmentation pathways
Ion-neutral reactions
Reaction rates
Determination of ionization
and appearance energies
Gas-phase basicities
Proton affinities

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GAS PHASE ION CHEMISTRY

Mass spectrometry aims to identify a compound from the molecular or
atomic mass(es) of its constituents.

Mass alone can be sufficient for the identification of elements and the
determination of the molecular formula of an analyte.

Relative abundance of isotopologs helps to decide which elements

contribute to such a formula and to estimate the number of atoms of a
contributing element.

Under the conditions of certain mass spectrometric experiments,

fragmentation of ions can deliver information on ionic structure. Thus, MS
elucidates the connectivity of atoms within smaller molecules, identifies
functional groups, determines the number and eventually the sequence of
constituents of macromolecules, and in some cases even yields their three-
dimensional structure
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ISOTOPES AND ISOTOPIC

DISTRIBUTIONS

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“Two very significant discoveries were

due to MS studies. First, J.J. Thomson
found that neon consisted of a
mixture of two different isotopes
(masses 20 and 22). This observation of
the existence of stable isotopes is
perhaps the greatest achievement
that can be claimed by MS.

Second, F.W. Aston observed that the

masses of all isotopes are not simple
multiples of a fundamental unit, but
rather they are characterized by a
mass defect; i.e., isotopes do not have
integral masses.”

Robert W. Kiser, The Introduction to Mass Spectrometry

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ISOTOPES IN MS

• Separation of charged species

by m/z ratio

• Molecular mass, calculated

using relative atomic masses, A= p+ + neutrons
is not observed in MS.

• MS does not separate by Z=Number of protons

elements but by isotopes.

Isotopes: = Z ≠ A
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Harmon R.S. (2018) Atomic Number, Mass Number, and Isotopes. In: White W.M. (eds) Encyclopedia of Geochemistry. Encyclopedia of Earth Sciences Series. Springer.

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Monoisotopic elements (20)

(A or X): Only one naturally occurring stable isotope (19F,23Na,31P,127I)

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Di-isotopic elements
(A+1 or X+1): H (1H:2H), C (12C:13C), N (14N:15N)
(A+2 or X+2): Cl (35Cl:37Cl), Br (79Br:81Br), Cu (63Cu: 65Cu), O, S, Si

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Di-isotopic elements
(A-1 or X-1): Li (6Li:7Li), B (10B:11B) and V (50V:51V)

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Poly-isotopic elements
Most elements consist of three or more isotopes with wide variety of
isotopic distributions.

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IUPAC Periodic Table

IUPAC of the Table
Periodic Isotopes
of the Isotopes
isotope mass number
Element Background Color Key element name cadmium (number of protons +
106 108 neutrons)

Cd
Standard atomic weights are the best estimates by IUPAC of atomic weights that are found in normal materials, which are terrestrial
materials that are reasonably possible sources for elements and their compounds in commerce, industry, or science. They are element symbol 113
113
116
116 .
, black indicates
114 isotope is stable
determined using all stable isotopes and selected radioactive isotopes (having relatively long half-lives and characteristic terrestrial
atomic number . red indicates
48 110
,

isotopic compositions). Isotopes are considered stable (non-radioactive) if evidence for radioactive decay has not been detected (number of protons) isotope is radioactive
experimentally.
111 isotopic abundance
1 standard 112 (mole fraction 18
Element has two or more isotopes that are used to determine its standard atomic weight. The isotopic abundances and of isotope)
hydrogen atomic weight 112.414(4) helium
atomic weights vary in normal materials. These variations are well known, and the standard atomic weight is given as lower
H He
{

and upper bounds within square brackets, [ ]. uncertainty in last digit (112.414 ± 0.004)
2 3
1 2
Element has two or more isotopes that are used to determine its standard atomic weight. The isotopic abundances and
4
1 atomic weights vary in normal materials, but upper and lower bounds of the standard atomic weight have not been assigned
[1.007 84, 1.008 11] 2 by IUPAC or the variations may be too small to affect the standard atomic weight value significantly. Thus, the standard 13 14 15 16 17 4.002 602(2)
lithium beryllium atomic weight is given as a single value with an uncertainty that includes both measurement uncertainty and uncertainty due boron carbon nitrogen oxygen fluorine neon
Li 6 Be 9
to isotopic abundance variations.
B 10 C 13 N 15 O 17
18 F 19 Ne 22
21

3 4 5 6 7 8 9 10
Element has only one isotope that is used to determine its standard atomic weight. Thus, the standard atomic weight is
7
invariant and is given as a single value with an IUPAC evaluated uncertainty. 11 12 14 16
20
[6.938, 6.997] 9.012 1831(5) [10.806, 10.821] [12.0096, 12.0116] [14.006 43, 14.007 28] [15.999 03, 15.999 77] 18.998 403 163(6) 20.1797(6)
Element has no standard atomic weight because all of its isotopes are radioactive and, in normal materials, no isotope
sodium magnesium occurs with a characteristic isotopic abundance from which a standard atomic weight can be determined.
aluminium silicon phosphorus sulfur chlorine argon
Mg
33 36

Na 23 26
(aluminum)
27 Si 29 P 31 S 34
Cl Ar 36
38

Al
25 30 37
11 12 14 15 16 17 18
13 35
24 28 32 40
22.989 769 28(2) [24.304, 24.307] 3 4 5 6 7 8 9 10 11 12 26.981 5385(7) [28.084, 28.086] 30.973 761 998(5) [32.059, 32.076] [35.446, 35.457] 39.948(1)
potassium calcium scandium titanium vanadium chromium manganese iron cobalt nickel copper zinc gallium germanium arsenic selenium bromine krypton
K Kr
42 48

Ca Sc Ti V Cr Mn Fe Co Ni Cu Zn Ga Ge As Se Br
61 70 74 78
40 44 43 57 64
46 47 49 53 50 54 58 62 67 76 82 77 80
41 46 45 55 59 60 68 70 75 83
50 50 54 65 71 73 76 81
19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 82

78
66 72 86
40 48 51 52 56 58 63 64 69 74 80 79
39 84

39.0983(1) 40.078(4) 44.955 908(5) 47.867(1) 50.9415(1) 51.9961(6) 54.938 044(3) 55.845(2) 58.933 194(4) 58.6934(4) 63.546(3) 65.38(2) 69.723(1) 72.630(8) 74.921 595(6) 78.971(8) [79.901, 79.907] 83.798(2)
rubidium strontium yttrium zirconium niobium molybdenum technetium ruthenium rhodium palladium silver cadmium indium tin antimony tellurium iodine xenon 124

Pd Sn Te Xe
106 108 122 123

Rb Sr Y Zr Nb Mo Tc Ru Rh Ag Cd In Sb I
84 102 112 114 130 128 126
94 100 96 109 116 113 122 115 123 124 120
86 87 89 92 91 93 97 99 103 104 113 125 127
87 96 98 114 124 136
98 106 120 130 132
37 38 39 40 41 42
100
43 44 45 46 110
47 48 110 49 50 117
51 52 126
53 54 134
94 119
92 101
131
85 88 105 111
90 96 102 116 128
95 104 108 107 112 115 118 121 129
85.4678(3) 87.62(1) 88.905 84(2) 91.224(2) 92.906 37(2) 95.95(1) 101.07(2) 102.905 50(2) 106.42(1) 107.8682(2) 112.414(4) 114.818(1) 118.710(7) 121.760(1) 127.60(3) 126.904 47(3) 131.293(6)
caesium barium 134 hafnium tantalum tungsten rhenium osmium iridium platinum gold mercury thallium lead bismuth polonium astatine radon
57 - 71
Ba Hf Ta W Re Os Ir Hg
130 174 186 184

Au Tl Pb Bi Po At Rn
180
(cesium)
Pt
135 132 187 192 190 196
176 207
133 183 198 197 198 204 203 204 209

Cs
136 179 180 180 185 188 191 202
184 192 195
56 137
lanthanoids 72 73 74 75 76 77 196
79 80 201 81 82 83 84 85 86
177 182
189
78 206
55 181
187 193 199 205 208
138 190 194
178 186 200
132.905 451 96(6) 137.327(7) 178.49(2) 180.947 88(2) 183.84(1) 186.207(1) 190.23(3) 192.217(3) 195.084(9) 196.966 569(5) 200.592(3) [204.382, 204.385] 208.980 40(1)
francium radium 89 - 103 rutherfordium dubnium seaborgium bohrium hassium meitnerium darmstadtium roentgenium copernicium ununtrium flerovium ununpentium livermorium ununseptium ununoctium
Fr Ra Rf Db Sg Bh Hs Mt Ds Rg Cn Uut Fl Uup Lv Uus Uuo
87 88 actinoids 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118

lanthanum cerium praseodymium neodymium promethium samarium europium gadolinium terbium dysprosium holmium erbium thulium ytterbium lutetium
La Ce Nd Sm Eu Gd Dy Er Yb Lu
136 138 158 156

Pr Pm Tb Ho Tm
144 154 152 164 162 168
160 170
142 141 148 150 150 155 159
161
165 170 169 176 174 176
138 142 152 151 158 164 166
57 58 59 60 145
61 62 148
63 64 65 66 67 68 69 70 171
71
143 157 167
149
163
139 153 173 175

https://applets.kcvs.ca/IPTEI/IPTEI.html
146 154 156 160
140 144 147 162 172
168
138.905 47(7) 140.116(1) 140.907 66(2) 144.242(3) 150.36(2) 151.964(1) 157.25(3) 158.925 35(2) 162.500(1) 164.930 33(2) 167.259(3) 168.934 22(2) 173.054(5) 174.9668(1)
actinium thorium protactinium uranium neptunium plutonium americium curium berkelium californium einsteinium fermium mendelevium nobelium lawrencium
Ac Th Pa U Np Pu Am Cm Bk Cf Es Fm Md No Lr
234
235
231
230
89 90 91 92 93 94 95 96 97 98 99 100 101 102 103

https://www.ciaaw.org/pubs/Periodic_Table_Isotopes.pdf 232

232.0377(4) 231.035 88(2) 238.028 91(3)

238

International Union of Pure and Applied Chemistry (IUPAC) Project 2007-038-3-200, "Development of an isotopic periodic table for the educational community" October 1, 2013 ww w.ciaaw.org

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https://phet.colorado.edu/sims/html/isotopes-and-atomic-mass/latest/isotopes-and-atomic-mass_en.html

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Atomic, Molecular, and Ionic Mass

Nominal mass: Approximate mass, sum up integer masses of the

elements.
CO2 = 12u + 2x(16u)= 44 u
Integer mass of most abundant naturally occurring isotope.

Isotopic mass:
Exact mass of the isotope (very close but not equal to the nominal
mass). C is the only exception (12C, isotopic mass 12.000000u)

Unified atomic mass [u] 1/12 of 12C nuclide

1[u]=1.66055 x 10-27 kg = 1 Da (1 dalton)
1 g = NA x u (or Da)

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Atomic, Molecular, and Ionic Mass

Relative Atomic Mass (atomic weight): Weighted average of the
naturally occurred isotopes of an element (Periodic Table).
𝒊 𝒊
35Cl (34.968853u / 75.78%)
𝑴𝒓 = $ 𝑨𝒊 𝒙 𝒎𝒊 #$ 𝑨𝒊 37Cl (36.965903u / 24.22%)
Mr Cl = 35.4528 u
𝒊%𝟏 𝒊%𝟏
Relative Molecular Mass (atomic weight): relative atomic masses of
the elements contributing to the empirical formula (not useful in MS)
Exact Ionic Mass:
M+. : Monoisotopic mass minus the mass of the electron
M-.: Monoisotopic mass plus the mass of the electron (s)

e.g. CO2+. = 12.000000u + 2x15.994915 u – 0.0000548 u = 43.989282 u

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Atomic, Molecular, and Ionic Mass

Sucrose: C12H22O11

Monoisotopic mass: 12C121H2216O11

(12 x12.0)+(22x1.007825)+(11x15.994915)= 342.116215 u
Nominal mass: 12C121H2216O11
(12 x12.0)+(22x1.0)+(11x16)= 342 u
Average mass: C12H22O11
(12x12.0107)+(22x1.00794)+(11x15.9994) = 342.29648 u
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I Patterns
ERPRINTS

Sucrose C12H22O11
12C 13C 1H 16O 18O Isotopolog Nom Isotop mass Abund%
mass
12 0 22 11 0 12C 13C 1H 16O 18O 342 342.116215
12 0 22 11 0 100,00
11 1 22 11 0 12C 13C 1H 16O 18O 343 343.119570
11 1 22 11 0 13,39
12 0 22 10 1 12C 13C 1H 16O 18O 344 344.120460
12 0 22 10 1 2,26
10 2 22 11 0 12C 13C 1H 16O 18O 344 344.122925
10 2 22 11 0 0,827
11 1 22 10 1 12C 13C 1H 16O 18O 345 345,123815 0,302
11 1 22 11 1

9 3 22 11 0 12C 13C 1H 16O 18O 345 345,126280 0,037

9 3 22 11 0

10 2 22 10 1 12C 13C 1H 16O 18O 346 346,127170 0,019

10 2 22 10 1

12 0 22 9 2 12C 13C 1H 16O 18O 346 346,124705 0,023

12 0 22 9 2

8 4 22 11 0 12C 13C 1H 16O 18O 346 346,129635 0,002

8 4 22 11 0

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I Patterns
ERPRINTS
Sucrose C12H22O11

Monoisotopic mass

A mass spectrum
superimposes the mass Isotopic ions
spectra of all isotopic (isotopologs)
species involved
M+1
M+2
Isotopic patterns are ideal M+3
sources of analytical
information

https://www.sisweb.com/mstools/isotope.htm
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I Patterns Calculation
ERPRINTS
1000 CH4 molecules.
11 molecules 13C instead of 12C
989 molecules 12CH4
Ratio (r) m/z 17 to m/z 16 = 1.1/98.9
𝒄
𝒓 = (𝟏𝟎𝟎%𝒄) c= 13C abundance

𝟏𝟎𝟎 − 𝒄 𝒘
Probability to have only 12C in a molecular 𝑷𝑴 =
ion M consisting of w carbons 𝟏𝟎𝟎
𝒘
𝒄 𝟏𝟎𝟎 − 𝒄
Probability of having exactly one 13C atom 𝑷𝑴)𝟏 = 𝒘
in an ion with w carbons 𝟏𝟎𝟎 − 𝒄 𝟏𝟎𝟎
𝑷𝑴)𝟏 𝒄
Ratio PM+1/PM =𝒘
𝑷𝑴 𝟏𝟎𝟎 − 𝒄
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I Patterns Calculation
ERPRINTS

Ratio probability of two 13C atoms in 𝑷𝑴)𝟐 𝒘 (𝒘 − 𝟏)𝒄𝟐

an ion with w carbons =
𝑷𝑴 𝟐(𝟏𝟎𝟎 − 𝒄)𝟐

Monoisotopic Mass: 720 Monoisotopic Mass: 1080 Monoisotopic Mass: 2160

Chemical Formula: C60 Chemical Formula: C90 Chemical Formula: C180
Average Mass: 720.6439 Average Mass: 1080.9662 Average Mass: 2161.9322

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I Patterns Calculation
ERPRINTS

The isotopic pattern of a complex

molecule is calculated by

1. Dividing the molecule into submolecules for each element

2. Deriving the isotopologues within these submolecules
3. Combining these subisotopologues to the exact isotopologue
probabilities and masses of the full molecule
4. Convoluting to measurable spectra at a given resolution by use
of peak shape functions
Accelerated Isotope Fine Structure Calculation Using Pruned Transition Trees, Anal. Chem. 2015, 87, 5738−5744 , DOI: 10.1021/acs.analchem.5b00941

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I Patterns Calculation
ERPRINTS

Melittin is the main component (40–60% of the dry

weight) and the major pain producing substance of
honeybee (Apis mellifera) venom . Melittin is a
basic peptide consisting of 26 amino acids.

C131H230N39O31
C131 O31
H230 N39

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I Patterns Calculation C131H230N39O31

ERPRINTS

C131 O31
H230 N39

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I Patterns Calculation
ERPRINTS
Binomial approach for di-isotopic elements
𝒏(𝒏 − 𝟏)𝒂𝒏%𝟐 𝒃𝟐 𝒏(𝒏 − 𝟏)(𝒏 − 𝟐)𝒂𝒏%𝟑 𝒃𝟑
(𝒂 + 𝒃)𝒏 = 𝒂𝒏 + 𝒏𝒂𝒏%𝟏 𝒃 + + + …
𝟐! 𝟑!
a,b = isotopic abundances of both isotopes.
n = number of this species in the molecule.
Example: C9N3Cl3
Cl (35: 0.7578; 37: 0.2422)
(𝟎. 𝟕𝟓𝟕𝟖 + 𝟎. 𝟐𝟒𝟐𝟐)𝟑 = 𝟎. 𝟕𝟓𝟕𝟖𝟑 + 𝟑(𝟎. 𝟕𝟓𝟕𝟖)𝟐 𝟎. 𝟐𝟒𝟐𝟐 + 𝟑 (𝟎. 𝟕𝟓𝟕𝟖)𝟏 (𝟎. 𝟐𝟒𝟐𝟐)𝟐 + 𝟎. 𝟐𝟒𝟐𝟐𝟑
(𝟎. 𝟕𝟓𝟕𝟖 + 𝟎. 𝟐𝟒𝟐𝟐)𝟑 = 𝟎. 𝟒𝟑𝟓 ∶ 𝟎. 𝟒𝟏𝟕 ∶ 𝟎. 𝟏𝟑𝟑 ∶ 𝟎. 𝟎𝟏𝟒 = 𝟏𝟎𝟎 ∶ 𝟗𝟓. 𝟗 ∶ 𝟑𝟎. 𝟕 ∶ 𝟑. 𝟑

C (12: 0.989; 13: 0.11)

(𝟎. 𝟗𝟖𝟗 + 𝟎. 𝟏𝟏)𝟗 = 𝟎. 𝟗𝟖𝟗𝟗 + 𝟗(𝟎. 𝟗𝟖𝟗)𝟖 𝟎. 𝟎𝟏𝟏 + 𝟑𝟔 (𝟎. 𝟗𝟖𝟗)𝟕 (𝟎. 𝟎𝟏𝟏)𝟐 + 𝟖𝟒(𝟎. 𝟗𝟖𝟗)𝟔 𝟎. 𝟎𝟏𝟏 𝟑

(𝟎. 𝟗𝟖𝟗 + 𝟎. 𝟎𝟏𝟏)𝟗 = 𝟎. 𝟗𝟎𝟓: 𝟎. 𝟎𝟗𝟎𝟔: 𝟎. 𝟎𝟎𝟒𝟎𝟑: 𝟎. 𝟎𝟎𝟎𝟏𝟎𝟒 = 𝟏𝟎𝟎 ∶ 𝟏𝟎. 𝟎𝟏: 𝟎. 𝟒𝟒𝟓: 𝟎. 𝟎𝟏𝟏𝟓

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I Patterns Calculation
Example: C9N3Cl3
C (12: 0.989; 13: 0.11) Cl (35: 0.7578; 37: 0.2422)
(𝟎. 𝟗𝟖𝟗 + 𝟎. 𝟎𝟏𝟏)𝟗 = 𝟏𝟎𝟎 ∶ 𝟏𝟎. 𝟎𝟏: 𝟎. 𝟒𝟒𝟓: 𝟎. 𝟎𝟏𝟏𝟓 (𝟎. 𝟕𝟓𝟕𝟖 + 𝟎. 𝟐𝟒𝟐𝟐)𝟑 = 𝟏𝟎𝟎 ∶ 𝟗𝟓. 𝟗 ∶ 𝟑𝟎. 𝟕 ∶ 𝟑. 𝟑

M M+1 M+2 M+3 M M+1 M+2 M+3 M+4 M+5 M+6

12C 1x 13C 2x 13C 3x 13C 35Cl 1x 37Cl 2x 37Cl 3 x 37Cl

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Isotopic Pattern Calculation

12C35Cl 1 x 37Cl
2 x 13C

E M M+1 M+2 M+3 M+4 M+5 M+6

m/z 255 256 257 257 259 260 261
C9 100 10 0.445 0.0115 - - -
Cl3 100 - 95.9 - 30.7 - 3.3
C9Cl3 100 10 96 0.0115 30.7 - 3.3
Teor 100 10.8 97.7 10.5 32 3.4 3.5

2 x 37Cl
4 x 13C

1 x 13C

3 x 37Cl

M M+1 M+2 M+3 M+4 M+5 M+6

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I Patterns Calculation
ERPRINTS

Polynomial approach for poly-isotopic elements

(a1 + a2 + a3 + …)m + (b1 + b2 + b3 + …)n + (c1 + c2 + c3 + …)o + …

ai : Isotopes of elements a (two or more).

m:n:o = number of this species in the molecule.

Stearic acid trichloromethylester = C19H35O2Cl3

𝐴23! + 𝐴24! 19 𝐵2" + 𝐵3" 35 𝐶25# + 𝐶26# + 𝐶27# 2 𝐷48!$ + 𝐷46!$ 3

19 35 2 3
2 2 3 2 = 1.297 x 1018

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NIST Chemistry WebBook

Isotope Calculator

CHEMCALC

ENVIPAT

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Isotopic Pattern Calculation

51

Measuring Mass
High Resolution and Accurate Mass
Mass accuracy depends on sufficiently resolved peaks
High resolution does not imply accurate mass

Exact Mass
Isotopic mass = exact mass of the isotope
Monoisotopic mass of a molecule or ion = exact mass
“No combination of elements in an empirical formula has the same CALCULATED
EXACT MASS as another one”

“At infinite mass accuracy it is possible to identify the empirical formula by mass
spectrometry alone”

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Measuring Mass
RESOLUTION Ability of an instrument to separate neighboring peaks

Isobaric ions Molecular Nominal Mass Monoisotopic

Ions mass
Ions having the N2+. 28u 28.00559 u
same nominal CO+. 28u 27.99437 u
mass C2H4 +.
28u 28.03075 u

RESOLUTION TO SEPARATE N2+. FROM CO+.

28
𝑅= = 2496
(28.00559 − 27.99437) Full width at
half maximum
RESOLUTION TO SEPARATE C2H4+. FROM CO+.
(FWHM)
28
𝑅= = 679
(28.03075 − 27.99437)

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Measuring Mass
Mass Accuracy
Difference between measured accurate mass and
calculated exact mass
Absolute units: u or mmu (10-3 u)
Relative units: ppm (absolute mass accuracy/mass)

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RINGS PLUS DOUBLE BONDS

Ni Number of atoms
Vi Valence of atoms. Monovalent (H,
F, Cl, Br, I). Divalent (O, S, Se).
Trivalent (N, P). Tetravalent (C, Si)

Restriction to formulas of the type

CcHhNnOo reduces the formula to:

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