EXPERIMENT NO.

MASS SPECTROMETRY

Name: Chariezza Le J. Sanglad

Student No.: 2015-00925

Name: Erlindo Regala

Student No.: 2015-01947

Name: Leziel Talpis

Student No.: 2015-01076

 ABSTRACT

Mass Spectrometry is an analytical spectroscopic tool that can determine the molecular

weight, molecular formula (HRMS), structure (from fragmentation fingerprint), isotopic

incorporation / distribution and protein sequence (MS-MS).

This experiment aimed to identify the compound or structure in the given mass spectrum

and to be able to interpret the fragments of the spectrum.

Based on the spectrum, given the fragments with m/z of 46, 45, 31,29, 15, the parent ion

peak which has a m/z of 46 did not match to 1-propanol, methanol and isopropanol. Therefore, it

corresponds only either formic acid or ethanol. The peak with 46 m/z is the parent molecular ion.

From this parent molecular ion, some of the fragments were formed by cleaving its particular

bonds. At 45 m/z, this is the molecular ion less with I hydrogen. At this fragmentation, the parent

ion has lost its hydrogen as a form of ion forming the fragment of [CH 3CH2O]+. One of the

properties of alcohols is that it can easily lose a hydrogen. This cleavage of CH3 was not possible

for formic acid because it has no CH3 group in its structure. At 29, this is the cleaving of the C-O

bond or the removal of the OH group forming the fragment [CH3CH2]+.. At fragment 45, the

removal of C-H bond occurred. At 29, the fragmentation for C-O bond and at 31, the removal of

C-C bond. In general, the spectrum clearly shows that the fragments correspond to the structure of

ethanol.
 INTRODUCTION

The determination of the structures of organic compounds is a vital part of chemistry in

general and analysis in particular. Mass Spectrometry is an analytical spectroscopic tool that can

determine the molecular weight, molecular formula (HRMS), structure (from fragmentation

fingerprint), isotopic incorporation / distribution and protein sequence (MS-MS). It is primarily

concerned with the separation of molecular (and atomic) species according to their mass.

The purposes of of this experiment are, to be able to identify the compound or structure in

mass spectrum and to be able to interpret the fragments of the spectrum.

The mass spectrometer is used to measure the masses and abundances of naturally

occurring isotopes. It is an instrument that generates ions from molecules and measures their

masses. A measure of how well a mass spectrometer separates ions of different mass is known as

resolution. In low resolution, it is capable of separating only ions that differ in nominal mass; that

is ions that differ by at least 1 or more atomic mass units while in high resolution, it refers to

instruments capable of separating ions that differ in mass by as little as 0.0001 atomic mass unit.

INPUT IONISER ELECTRIC MAGNETIC ELECTRIC

FIELD FIELD FIELD

DETECTOR
Figure 1.Block Diagram for Mass Spectrometer Instrument

When an atom or molecule is hit by high-energy electron, it will ionize. The electron is

deflected but transfers much of its energy to the molecule. This energy-rich species ejects an

electron forming a positively charged, odd-electron species called the molecular ion. Molecular
ion passes between poles of a magnet and is deflected by magnetic field. If the only ion that is

present is the molecular ion, mass spectrometry provides a way to measure the molecular weight

of a compound and is often used for this purpose. However, the molecular ion often fragments to

a mixture of species of lower m/z. The molecular ion dissociates to a cation and a radical. Usually

several fragmentation pathways are available and a mixture of ions is produced. mixture of ions of

different mass gives separate peak for each m/z. Intensity of peak proportional to percentage of

each ion of different mass in mixture. The separation of peaks depends on relative mass.

A molecular ion (M) is a radical cation formed by removal of a single electron from a

parent molecule in a mass spectrometer. For some purposes, it does not matter which electron is

lost; radical cation character is delocalized throughout the molecule; therefore, the molecular

formula of the parent molecule is written in brackets with plus sign to show that it is a cation and

a dot to show that it has an odd number of electrons. Fragment ions rationalize these in terms of

the structure. It identifies as many as possible, in terms of the parent structure. Generally, simply

derived from the molecular ion or, in a simple fashion from a significant higher mw fragment.

Fragments will make chemical se nse structure of the compound. Fragmentation are governed by

product ion stability consideration, octet rule, resonance delocalization, polarizability and

hyperconjugation as well as electronegativity.

The ionising electron beam can fragment the molecule by cleaving particular bonds. The

molecular fragmentation peaks in the mass spectrum can give clues as to the sub-structure of the

molecule. The peak height is the relative abundance of each species. The most intense peak (called

the Base peak) is assigned a relative abundance of 100. The parent ion peak (i.e. the singly charged

ion of the test molecule denoted by M + located at m/e of 114 in the spectrum above) is often too

weak to be seen in the high energy spectra that are necessary to produce a fragmentation pattern.
 METHODOLOGY

In mass spectrometry, a sample was introduced in an input and then bombarded in

an ionizer with a beam of electrons. The ions were then accelerated by an electric field and

deflected in a curve by a further electric field. The various ions were linearly accelerated then

passed through a magnetic field to curve their flight path. The ions were passed again through

another electric field to accelerate into the detector. By varying the electric and magnetic fields,

ions of different mass can be made to enter the detector. Furthermore, there was a pump that keeps

the interior of an instrument under a high vacuum, this was used to allow the ions which were

moving at speed of many kilometer per second, a free path without collisions with air molecules.

In addition, less volatile sample were dissolved in a solvent such as dichloromethane which then

evaporates. The sample tube was placed in a probe which was then inserted into the instrument

with an airlock so that the vacuum in an instrument is maintained. The volatile sample was

vaporized at low pressure inside the instrument but the probe can be heated to vaporize less volatile

sample. The electron streamed out from a heated cathode towards an anode which was held at a

potential of around plus 70V relative to the cathode. The positive ions were formed in ionization

chamber, these were formed when high energy electron strike the sample molecule. The positive

ion beam was passed between two charged curve plates which were then deflected as it passes

through the magnetic field. The beam was deflected again by another set of charged plates which

directed into the detector. The amount of deflection by the magnetic field depends on the mass of

an ion wherein heavier ions was not deflected sufficiently in a magnetic field to reach the detector

while light ions was deflected too much.

 RESULTS AND DISCUSSION

The figure below shows the given spectrum for the identification of a compound.

COMPOUNDS Formic acid 1-propanol ethanol methanol Isopropyl

alcohol
MW(g/mol) 46.0262 60.0977 46.0702 32.0428 60.0977

Figure 1. Masss Spectrum of Ethanol

Molecular ion peak/fragments m/z values structure

[CH3CH2OH]+. 46 H H

[CH3CH2O]+. 45 H C C O H

[CH2OH]+. 31 H H

[CH3CH2]+. 29

The mass spectrum above may correspond to one of the compounds in the table.

There were five compounds shown in the table namely: Formic acid(46.0262g/mol), 1-

propanol(60.0977g/mol), ethanol(46.0702g/mol), methanol(32.0428g/mol), and isopropyl

alcohol(60.0977g/mol). Looking at the spectrum, it can be seen that the parent molecular ion peak

produced from the collision of the sample with a beam of electrons has a mass to charge ratio of

46. The given spectrum did not match to 1-propanol and isopropyl alcohol for the reason that their

molecular weights were too high. In the same manner the methanol compound did not match with

the given spectrum because it has too low molecular weight. Therefore, this spectrum corresponds

only either to formic acid or ethanol since these two compounds have a molecular weight which

were close enough to the m/z value of the parent molecular ion.

At high energies, the electron beam can fragment the molecule by cleaving particular

bonds. There were four major peaks in the spectrum: 46, 45, 31, and 29. The peak with 46 m/z is

the parent molecular ion. From this parent molecular ion, some of the fragments were formed by

cleaving its particular bonds. At 45 m/z, this is the molecular ion less with I hydrogen. At this

fragmentation, the parent ion has lost its hydrogen as a form of ion forming the fragment of

[CH3CH2O]+. One of the properties of alcohols is that it can easily lose a hydrogen. Formic acid

is a carboxylic acid and ethanol is an alcohol so, this is one of the clues that this spectrum belongs

to ethanol. Looking further to the highest peak which is called the base peak, it was observed that

this is when the cleaving of the C-C bond happened or the removal of CH3 from the parent ion

results in the formation of the fragment [CH2OH]+. This cleavage of CH3 was not possible for

formic acid because it has no CH3 group in its structure. At 29, this is the cleaving of the C-O bond

or the removal of the OH group forming the fragment [CH3CH2]+. The cleavage of hydrogen at 45

and the cleavage of CH3 at peak 31 validate that this spectrum is for ethanol.
 CONCLUSION

This experiment aimed to identify which among the five compounds namely: formic acid,

1-propanol, ethanol, methanol, and isopropyl alcohol corresponds to the given mass spectrum. This

aimed also to interpret the fragments of the spectrum.

The results showed the structure of ethanol matches with the fragments of the spectrum.

The cleavage of hydrogen at fragment 45 and the removal of CH3 at fragment 31 which can be

happened only for ethanol because it is an alcohol and can be deprotonated by an attack of

electrons. Formation of fragment at 31 is not possible for formic acid because it does not have a

methyl group in its structure.

 REFERENCES

a.) Mass Spectrum-Compound-PubChem.ncbi.nlm.nih.gov

b.) R. García and A. P. Báez (2012). MS, Dr. Muhammad Akhyar Farrukh (Ed.), ISBN: 978-

953-307-817-5

c.) Wiley; Mass Spectrometry: Principles and Application, 3rd Edition

d.) Mass Spectrometry- A Textbook about Book, 3rd ed.,

e.) Introduction to Mass Spectrometry- A Textbook by Edmond de Hoffman

 APPENDIX

COMPOUNDS Formic acid 1-propanol ethanol methanol Isopropyl

alcohol
MW(g/mol) 46.0262 60.0977 46.0702 32.0428 60.0977

Molecular ion peak/fragments m/z values structure

[CH3CH2OH]+. 46 H H

[CH3CH2O]+. 45 H C C O H

[CH2OH]+. 31 H H

[CH3CH2]+. 29