The document discusses hyphenated analytical techniques, emphasizing their role in combining separation and detection methods to enhance analytical capabilities. It outlines various techniques such as GC-MS, LC-MS, ICP-MS, and MS-MS, detailing their working principles and advantages. The advantages include fast and accurate analysis, higher automation, and improved sample throughput.

1. HYPHENATED TECHNIQUES
Presented By : Biji Saro Varghese
M.Sc. II Sem-IV
1
RASHTRASANT TUKADOJI MAHARAJ NAGPUR UNIVERSITY
P. G. T. DEPARTMENT OF CHEMISTRY
7/10/2019

2. CONTENTS
• INTRODUCTION
• GC-MS
• LC-MS
• ICP-MS
• MS-MS
7/10/2019 2

3. INTRODUCTION
Hyphenated technique is a combination or coupling
of two analytical techniques with the help of proper
interface.
The hyphenated technique is developed from the
coupling of a separation technique and detection
technique.
The term “hyphenation” was first adapted by
Hirschfeld in 1980 to refer to the on-line
combination of a separation technique and one or
more spectroscopic detection techniques.
7/10/2019 3

4. List of hyphenated techniques
• GC-MS
• LC-MS
• LC-NMR
• EC-MS
• CE-MS
• ICP-MS
• GC-IR
• LC-MS-MS
• MS-MS
• GC-MS-MS
• GC-NMR
• GC-AES
• ICP-AAS7/10/2019 4

5. Fig: Block diagram of Hyphenated technique
7/10/2019 5

6. Gas
chromatography
Mass
spectrometry
GC-MS
Separates
mixture of
components
into individual
Identifies
molecules
based on their
mass
A chemical analysis
technique
combining two
instruments to
provide for powerful
separation &
identification.
GC-MS
7/10/2019 6

7. • Gas chromatography–mass spectrometry(GC-MS, or alternatively
HPLC-MS) is an advanced analytical instrumental technique
that combines the physical separation capabilities of gas
chromatography with the mass analysis capabilities of mass
spectrometer
• It is one of the most accurate and efficient tools for analyzing
volatile organic samples.
• After separation in the GC column, analyte species have to be
transported to the mass spectrometer to be ionized, mass filtered
and detected.
• The column outlet needs to be connected to the ion source of the
mass spectrometer and different strategies had been implemented
which needs to fulfill the following conditions:
o Analyte must not condense in the interface.
o Analyte must not decompose before entering the mass
spectrometer ion source.
o The gas load (dictated by the mobile phase gas flow rate) entering
the ion source must be within the pumping capacity of the mass
spectrometer.
7/10/2019 7

8. GC-MS Coupling
Direct Introduction
Jet separator
Bieman Concentrator
Fig : Diagrammatic representation of GC-MS
7/10/2019 8

9. Types of interfaces:
Direct Introduction
7/10/2019 9

10. Direct Introduction
GC-MS systems use capillary columns & fused silica tubing
permits an inert, high efficiency, direct transfer between the
2 systems.
Flow rates is 5ml/min.
sdfssdhj
fh
7/10/2019 10

11. Jet Separator
The jet separator consist of two glass tubes
aligned with a Small distance between them.
Carrier gas entering from the GC column is
pumped away by a separate vacuumed system.
The larger sample molecules maintain their
momentum &pass preferentially in to the second
capillary. Sample enrichment occurs & the initial
atmospheric pressure is reduced.
sdfssdhj
fh
7/10/2019 11

12. Watson-Biemann effusion separator
It consists of a sintered glass tube . The carrier
gas usually helium passes preferentially through
the sintered glass tube and the effluent is
concentrated . The gas flow rates in the order of
20-60ml/min.
sdfssdhj
fh
7/10/2019 12

13. LC-MS
• Liquid chromatography-mass spectrometry is the technique
which performs separation by liquid chromatography and
mass analysis with the help of the mass spectrometry.
• Electrospray needle is used as bridge to connect the liquid
chromatography with that of the mass spectrometry. LC-MS
is mainly separated into the three parts-
 Chromatography
In liquid chromatography separation is performed which is
detected with the help of Photo diode Array. These separated
components then transferred to the Interface.
 Interface
In interface the liquid is volatilized and transferred to the MS.
 Spectrometry
With the help of various ionization techniques the compound
is ionized and then it is analyzed by mass analyzer.
7/10/2019 13

14. 7/10/2019 14

15. Fig: Schematic diagram of LC-MS
7/10/2019 15

16. ICP-MS
• Inductively coupled plasma mass
spectrometry (ICP-MS) is a type of mass
spectrometry which is capable of
detecting metals and several non-metals at
concentrations as low as one part in 1015 (part
per quadrillion, ppq) .
• This is achieved by ionizing the sample
with inductively coupled plasma and then using
a mass spectrometer to separate and quantify
those ions.
7/10/2019 16

17. Fig: Schematic representation of ICP-MS7/10/2019 17

18. MS-MS Tandem
In a tandem mass spectrometer, ions are formed in
the ion source and separated by mass-to-charge in
the first stage of mass spectrometry (MS1). Ions of
a particular mass-to-charge ratio (precursor ions)
are selected and fragment ions (product ions) are
created by collision-induced dissociation, ion-
molecule reaction, photo dissociation, or other
process. The resulting ions are then separated and
detected in a second stage of mass spectrometry
(MS2). A common use is for analysis of
biomolecules such as proteins and peptides.
7/10/2019 18

19. Fig :Diagrammatic representation of tandem mass spectrometry
7/10/2019 19

20. Advantages of hyphenated techniques
 Fast and accurate analysis.
 Higher degree of automation.
 Higher sample throughput.
 Better reproducibility.
 Reduction of contamination due to its
closed system.
 Separation and quantification achieved
at same time.
7/10/2019 20

21. References
• https://en.wikipedia.org/wiki/Tandem_mass_spectrometry
• https://en.wikipedia.org/wiki/Inductively_coupled_plasma_mass_spectrometry
• https://en.wikipedia.org/wiki/Liquid_chromatography%E2%80%93mass_spectrometry
• https://www.chromacademy.com/subscribe-warning.asp
1. Trace analysis by Susanta Lahiri, Narosa Publishing House
2. Web references
7/10/2019 21

22. THANK YOU
7/10/2019 22