ASSIGNMENT

On

HYPHENATED TECHNIQUES
By

KHAWAR KHALID
(Roll Number: 42)

Pharm-D (Final Year)

Federal Urdu University of Arts, Science &

Technology, Karachi

Assignment Submitted To

Sir Zahid
 HYPHENATED TECHNIQUES

DEFINITION AND BACKGROUND

Definition
“The term “Hyphenated” is basically a technique which is the combination of Chromatographic
and Spectroscopic methods to exploit the advantages of both.”

The term “Hyphenated” technique was first introduced by “Hirschfeld” refers to an online
combination of a chromatographic separation technique with a sensitive and element-specific
detector (usually an atomic absorption, emission or mass spectrometer).

“Hyphenation” terminology refers to integrated analytical systems that enable improved

performance and provide access to new applications. One of the most powerful hyphenated
techniques is LC-MS which is based on the coupling of a HPLC system with a Mass
spectrometer.

Examples:

Gas chromatography-mass spectrometry Gas chromatography-infrared spectroscopy

Liquid chromatography-mass spectrometry Liquid chromatography-NMR spectroscopy
Liquid chromatography-infrared Spectroscopy Capillary electrophoresis-mass spectrometry

HYPHENATED HPLC TECHNIQUES

Analytical HPLC systems are increasingly used in “hyphenated techniques” such as LC/NMR or
LC/MS where the substance is first separated with LC, and then introduced into the NMR or
mass spectrometer for more exquisite determination of molecular identification, structure,
function and dynamics.

LC/MS or LC/Mass Spectrometry: These systems combine the chemical separating power of
liquid chromatography with the ability of a mass spectrometer to selectively detect and confirm
molecular identity. MS is a highly selective technique that can literally find the "needle"
compound in a "haystack" sample. The technique also provides confirmatory information about
analyte identities. This qualitative analysis makes it possible to reconstruct an unknown
compound from MS data.

The Process: With LC/MS, liquid from the chromatograph is nebulized into
micro-droplets in the atmospheric interface between the LC and the mass
spectrometer. These droplets evaporate rapidly and release ionized analyte
molecules into the orifice that leads to the vacuum side of the interface.
Passing through additional stages of vacuum, ions are ultimately transferred
into the high vacuum region of the MS for separation based upon the ion's
mass and charge (degree of ionization). Further fragmentation of these ions
by MS/MS significantly increases the information about the molecule and the
selectivity of the detection.