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INTRODUCTION “Hyphenated Techniques” is “the combination of two separate analytical techniques via appropriate interface, usually with the backup of computer tying everything together”. The term hyphenated techniques ranges from the combination of spectroscopic techniques with chromatographic techniques or the coupling of separation techniques with spectroscopic detection techniques. Hyphenated techniques is used to analyzing complex mixture of peptides, separation of compound which contain ester, fatty acid, alcohol, terpenes. To Study the Principle, Instrumentation and Application of following:- Gas Chromatography-Mass spectrometry Ion Mobility Spectrometry-Mass spectrometry Capillary Electrophoresis-Mass spectrometry High Performance Liquid Chromatography –Mass spectrometry Liquid Chromatography-Mass spectrometry


AIM & OBJECTIVE To study theory and instrumentation of hyphenated Techniques. This helps in Biomarker identifications both for pharmacodynamic analysis and for mechanistic investigations of disease and processes. Detection of explosives analysis of nano particles. It suitable for separating nonvolatile species.

Gas Chromatography-Mass Spectroscopy: 

Gas Chromatography-Mass Spectroscopy

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Gas chromatography-mass spectroscopy (GC-MS):- It is method that combines the features of gas-liquid chromatography and mass spectroscopy to identify different substance within a test sample. Applications of GC-MS include drug detection, fire investigation, environmental analysis, explosives investigations, and identification of unknown samples. GC/MS can also be used in airport security to detect substance in luggage or on human beings. Additionally, it can identify trace elements in material that were previously thought to have disintegrated beyond identification. The component of gas chromatography as follows- 1. Carrier gas 2. Sample injection system 3. Columns 4. Detectors Detectors There are many detector which can be used in gas chromatography. Different detector will give different types of selectivity. A non-selective detector responds to all compounds expects the carrier gas, a selective detector responds to range of compounds except the carrier gas, a selectivity detectors responds to a range of compounds with a common physical or chemical property and a specific detectors responds to a single compound.

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Types Of Detector:- 1.Thermal Conductivity Detector (TCD) 2. Flame – Ionization Detector (FID) 3.Eelectron Capture Detector (ECD) Application:- 1.Enviourmental Monitoring and Cleanup- The cost of GC-MS equipment has decreased significantly, and the reliability has increased at the same time, which has contributed to have increased absorption in enviourmental studies. 2. Law Enforcement- GC-MS is increasingly used for detection of illegal narcotics, and may eventually supplant drug-sniffing dogs. 3. Medicine- In combination with isotopic labeling of metabolic compounds, the GC-MS is used for determining metabolic activity. 4. Food, Beverage and Perfume Analysis Foods and beverages contain numerous aromatic compounds, some naturally present in the raw material and some forming during processing.

Ion Mobility Spectroscopy-Mass Spectrometry: 

Ion Mobility Spectroscopy-Mass Spectrometry

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Ion Mobility Spectrometry –Mass Spectrometry :- The IMS-MS is composed of two major building blocks: the ion mobility spectrometer and the mass spectrometer. Whereas the ion mobility spectrometer is usually made of a drift region at atmospheric pressure or lower, the mass under a high vacuum. The component of IMS Ionization Analyzer Drift gas pressure Ion traps Applications :- The IMS-MS techniques can be used in proteonomics, for analyzing complex mixtures of peptides. Proteonomics for the analysis of peptides. Detection of chemical warfare agents. Detection of explosives analysis of nano particles.

Capillary Electrophoresis-Mass Spectrometry: 

Capillary Electrophoresis-Mass Spectrometry

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Capillary electrophoresis mass spectrometry (CEMS) :- It is an analytical chemistry technique formed by the combination of the liquid separation process of capillary electrophoresis with mass spectrometry. Ions are typically formed by electro spray ionization, but they can also be formed by matrix-assisted laser desorption ionization or other ionization techniques. It has applications in basic research in proteonomics and quantitative analysis of biomolecules as well as in clinical medicine. The component of capillary as follows :- Sample Introduction Detection:-a)Direct UV Detection b) Fluroscence Detection Detector:-a) Direct UV Vis Detector b) Photodiode Detector

High Performance liquid Chromatography-Mass Spectrometry: 

High Performance liquid Chromatography-Mass Spectrometry

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HIGH PERFORMANCE LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY:- One of the important analytical sy s tems is used in laboratories today is the coupling of HPLC (High performance liquid chromatography) with MS (Mass spectrometry).This systems is used in a wide range of situations which include the pharmaceutical, enviourmental, biological and food industries. The HPLC systems separates the components mixtures and The MS measures their molecular weights (relative molecular masses). This powerful technique is sometimes referred to by yet another abbreviation, LCMS-Liquid Chromatography Mass Spectrometry. The Components of HPLC systems:- Solvent Pump Detectors(Diode Array) Solvent Delivery System (Pump) Injector Column Sample

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Applications:- HPLC used to analytical separation. It is suitable for separating nonvolatile species. The wide variety of packing material allow the separation of most chemical species. The different types of detectors available permit the sensitive detection of most chemical types, and the accuracy and precision with which eluted substance may be quantified give analytical data of the highest caliber. The short column in routine use allow fast separation to take place, and often a complete separation of a complex mixture can be achieved within a few minutes. Microcrystalline packing material give excellent separation of similar substance.

Liquid Chromatography-Mass Spectrometry: 

Liquid Chromatography-Mass Spectrometry

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Liquid Chromatography-Mass Spectrometry:- Liquid chromatography-mass spectrometry(LC-MS, or alternatively HPLC-MS) is an analytical chemistry technique that combines the physical separation capabilities of liquid chromatography (or HPLC) with the mass analysis capabilities of mass spectrometry.LC-MS is a powerful technique used for many applications which has very high sensitivity and specificity. Generally its applications is oriented towards the specific detection and potential identification of chemicals in the presence of other chemicals (in a complex mixture). The component of mass spectrometer Inlet system Ion source Mass analyzer Ion traps Detector


Applications:- Pharmacokinetics LC-MS is commonly used in pharmacokinetics studies of pharmaceuticals. These studies give information about how quickly a drug will be cleared from the hepatic Blood flow, and organs of the body.MS is used for this due to high sensitivity and exceptional compared to UV (as long as the analyte can be ionized), and short analysis time. Drug development LC-MS is frequently used in drug development at many different stages including peptide Mapping, Glycoprotein Mapping, Natural Products Dereplication, Bioaffinity Screening, In vivo Drug Screening, Metabolite Stability Screening, Metabolite Identification, Impurity Identification, Degradant Identification, Quantitative Bioanalysis , and Quality control. Proteomics LC-MS is also used in the study of proteonomics where again components of a complex mixture must be detected and identified in some manner. Atom Probe An atom probe is an instrument that combines time-of –flight mass spectrometry and field ion microscopy (FIM) to map the location of individual atoms. Trace gas analysis Several technique use ions created in a dedicated ion source injected into s flow tube or a drift tube; selected ion flow tube (SIFT-MS), and proton transfer reaction (PTR-MS).


Conclusion The study of hyphenated technique based on separation of mixtures of compound. This technique is used to identify different substance within a test sample and also to detect and separate chemicals from solutions. The hyphenated techniques are GC-MS,IMS-MS,CE-MS,LC-MS. By using this technique, we identify drug at many different stages including Peptides mapping, Glycoprotein Mapping, Natural Products Dereplication, Bioafffinity Screening.


Reference A.H.Beckett , J. B. Stenlake,”Practical pharmaceutical chemistry”, CBS publications 4 th Edition, volume-2 1988, page no.157-166. A.P. Bruins. “Mass spectroscopy with ion sources operating at atmospheric pressure”. 1991, page no.53-77. Douglas A. Skoog , F. James Holler, Timothy A. Nieman . “ Principles of Instrumental Analysis”. 5 th edition, 2004 page no,725-739,781-787. Gurudeep R. Chatwal , Sham K. Anand . “ Instrumentation methods of chemical analysis”,Himalaya publication house, 5 th edition, 2002, page no.2.673-2.700,2.272-2.302 M.A Park, J.H Callahan and A.Vertes .” An inductive detector for time-of flight mass spectrometry”.1994,page no 317-322.

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