Hyphenated technique is a combination or coupling
of two analytical techniques with the help of proper
The hyphenated technique is developed from the
coupling of a separation technique and detection
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. • 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
• 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
• 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
10. Direct Introduction
GC-MS systems use capillary columns & fused silica tubing
permits an inert, high efficiency, direct transfer between the
Flow rates is 5ml/min.
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.
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
• 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-
In liquid chromatography separation is performed which is
detected with the help of Photo diode Array. These separated
components then transferred to the Interface.
In interface the liquid is volatilized and transferred to the MS.
With the help of various ionization techniques the compound
is ionized and then it is analyzed by mass analyzer.
• 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
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.
20. Advantages of hyphenated techniques
Fast and accurate analysis.
Higher degree of automation.
Higher sample throughput.
Reduction of contamination due to its
Separation and quantification achieved
at same time.