TOPIC: Mass spectrometry

Submitted by Submitted to
Sayma sadia Dr. S. M. Riajul Wahab
Id: 201811231003 Senior Lecturer
Batch: 31st Department of pharmacy
Department of pharmacy Bangladesh University
Bangladesh University
CONTENTS

1) Mass spectrometry Definition & Uses

2) How is mass spectrometry used in medicine?

3) Mass spectrometer

4) Components of a mass spectrometer

5) Inlet system

6) types of mass spectrometers are used

7) principle of mass spectrometry

8) Component of mass spectrometer

9) Analysis of Biomolecules using Mass Spectrometry

Mass spectrometry definition & uses

Mass spectrometry is an analytical technique that is used to measure

the mass-to-charge ratio of ions. The results are typically presented as a
mass spectrum, a plot of intensity as a function of the mass-to-charge
ratio.

Specific applications of mass spectrometry include

 Drug testing and discovery,

 Food contamination detection,
 Pesticide residue analysis,
 Isotope ratio determination,
 Protein identification, and
 Carbon dating.

These measurements can often be used to calculate the exact

molecular weight of the sample components as well.

Mass spectrometry used in medicine

Clinical mass spectrometry uses the mass spectrometry technology for
diagnostic purposes. Employed by medical labs, clinical mass
spectrometry is used to diagnose metabolism deficiencies, to
determine whether biomarkers or enzymes are present, and for
toxicology testing.
Types of mass spectrometers are used
analyzers
- MALDI-TOF
- ICP-MS
- DART-MS
- Secondary ion mass spectrometry (SIMS)
Types of ion detector
- Electron multiplier (EM)
- Faraday cup (FC)
- Photomultiplier conversion dynode
- Array detectors
Combining mass spectrometers with other techniques
- Gas chromatography mass spectrometry (GC-MS)
- Liquid chromatography mass spectrometry (LC-MS)
- Crosslinking mass spectrometry (XL-MS)
- Hydrogen-exchange mass spectrometry (HX-MS)
- Matrix-assisted laser desorption/ionization mass spectrometry
imaging (MALDI-MSI)
MASS SPECTROMETER
It is an instrument in which the substances in gaseous or vapor state is
bombarded with a beam of electrons, to form positively charged ions
(cations) which are further sorted according to their mass to charge
ratio to record their masses and relative abundances.

It is a sorted collection of the masses of all the charged molecular

fragments produced, the relative abundance of each is the
characteristics of every compound.

The mass spectrum can give detail information about composition of an

organic compound and the position of functional groups and is also
used for the determination of molecular weight.
COMPONENTS OF A MASS SPECTROMETER
The essential components of a mass spectrometer consist of:

 A sample inlet
 An ionization source
 A mass analyzer
 An ion detector
 Vacuum system
INLET SYSTEM:
The selection of a sample inlet depends upon the sample and the
sample matrix. Most ionization techniques are designed for gas phase
molecules so the inlet must transfer the analyte into the source as a gas
phase molecule. If the analyte is sufficiently volatile and thermally
stable, a variety of inlets are available. Gases and samples with high
vapor pressure are leaked directly into the source region by the help of
mercury manometer. Liquids and solids are usually heated to increase
the vapor pressure for analysis. Liquid samples are handled by
hypodermic needles injection through a silicon rubber dam If the
analyte is thermally labile (it decomposes at high temperatures) or if it
does not have a sufficient vapor pressure, the sample must be directly
ionized from the condensed phase.

SAMPLE INTRODUCTION METHODS:

1) Direct Vapor Inlet
2) Gas Chromatography
3) Liquid Chromatography
4) Direct Insertion Probe
5) Direct Ionization of Sample
PRINCIPLE OF MASS SPECTROMETRY
A mass spectrometer generates multiple ions from the sample under
investigation, it then separates them according to their specific mass-
to-charge ratio (m/z), and then records the relative abundance of each
ion type.
The first step in the mass spectrometric analysis of compounds is the
production of gas phase ions of the compound, basically by electron
ionization. This molecular ion undergoes fragmentation. Each primary
product ion derived from the molecular ion, in turn, undergoes
fragmentation, and so on. The ions are separated in the mass
spectrometer according to their mass-to-charge ratio, and are detected
in proportion to their abundance. A mass spectrum of the molecule is
thus produced. It displays the result in the form of a plot of ion
abundance versus mass-to-charge ratio. Ions provide information
concerning the nature and the structure of their precursor molecule. In
the spectrum of a pure compound, the molecular ion, if present,
appears at the highest value of m/z (followed by ions containing
heavier isotopes) and gives the molecular mass of the compound.
Components
The instrument consists of three major components:

1. Ion Source: For producing gaseous ions from the substance being
studied.
2. Analyzer: For resolving the ions into their characteristics mass
components according to their mass-to-charge ratio.
3. Detector System: For detecting the ions and recording the
relative abundance of each of the resolved ionic species.

In addition, a sample introduction system is necessary to admit the

samples to be studied to the ion source while maintaining the high
vacuum requirements (~10-6 to 10-8 mm of mercury) of the technique;
and a computer is required to control the instrument, acquire and
manipulate data, and compare spectra to reference libraries.

Figure: Components of a Mass Spectrometer

With all the above components, a mass spectrometer should always
perform the following processes:

1. Produce ions from the sample in the ionization source.

2. Separate these ions according to their mass-to-charge ratio in the
mass analyzer.
3. Eventually, fragment the selected ions and analyze the fragments
in a second analyzer.
4. Detect the ions emerging from the last analyzer and measure
their abundance with the detector that converts the ions into
electrical signals.
5. Process the signals from the detector that are transmitted to the
computer and control the instrument using feedback.

Analysis of Biomolecules using Mass

Spectrometry
Mass spectrometry is fast becoming an indispensable field for analyzing
biomolecules. Till the1970s, the only analytical techniques which
provided similar information were electrophoretic, chromatographic or
ultracentrifugation methods. The results were not absolute as they
were based on characteristics other than the molecular weight. Thus
the only possibility of knowing the exact molecular weight of a
macromolecule remained its calculation based on its chemical
structure.

The development of desorption ionization methods based on the

emission of pre-existing ions such as plasma desorption (PD), fast atom
bombardment (FAB) or laser desorption (LD), allowed the application of
mass spectrometry for analyzing complex biomolecules.