lc-ms ppt

Category: Education

Presentation Description

Introduction, Application&Principal Of LC-MS.


Presentation Transcript

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Prepared by: PATEL PRATIK L. (M.Pharm, P.C. 1st semester) Guided by: (1) Prof. Dr. G. Vidhyasagar (M.Pharm, Ph.D., PGDMM., DIH., FIC., FES., FABAP) (2) Zarna R. Dedania (M.Pharm, Q.A.) Department of P’ceutical Analysis, Veerayatan Institute of Pharmacy Mandvi, Kutch.

What is LC-MS?:

What is LC-MS? It is the combination of liquid chromatography and the mass spectrometry. In LC-MS we are removing the detector from the column of LC and fitting the column to interface of MS. In the most of the cases the interface used in LC-MS are ionization source.


PROBLEMS IN COMBINING HPLC AND MS HPLC Liquid phase operation 25 - 50 deg. C No mass range limitations Inorganic buffers 1 ml/min eluent flow is equivalent to 500 ml/min of gas MS Vacuum operation 200 - 300 deg. C Up to 4000 Da for quadrupole MS Requires volatile buffers Accepts 10 ml/min gas flow

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MOBILE PHASE :-The mobile phase is the solvent that moves the solute through out column. General requirements:- (1)low cost, uv transperancy,high purity. (2)low viscosity, low toxicity, non flammability. (3)non corrosive to LC system component. Solvent strength and selectivity:- it is the ability of solvent to elute solutes from a column.

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COlUMN :- Column type:- Specialized mode:- The use of di-functional or tri-functional silanes to create bonded groups with two or three attachement points leading to phases with higher stability in low or higher pH and lower bleed for LCMS Most widely used columns for LCMS are:- (1) fast LC column. the use of short column. (15-50mm) (2) Micro LC column. the use of large column. ( 20-150mm)

Sample preparation:-:

Sample preparation:- Sample preparation generally consists of concentrating the analyte and removing compounds that can cause background ion or suppress ionization. Example of sample preparation include:- (1) on –column concentration to increase analyte concentration. (2) desalting to reduce the sodium and potassium adduct formation that commonly occurs in electro spray. (3) filtration to separate a low molecular-weight drug from proteins in plasma, milk, or tissue.


Interface:- It is difficult to interface a liquid chromatography to a mass-spectrometer cause of the necessity to remove the solvent. The commnly used interface are:- (1) Electrospray ionization (ESI) (2) Thermospray ionization (TSI) (3) Atmospheric pressure chemical ionization (APCI) (4) Atmospheric pressure photoionization(APPI) (5) Partical beam ionization.

Electron spray ionization.:

Electron spray ionization.

Atmospheric pressure chemical ionization (APCI) :

Atmospheric pressure chemical ionization (APCI)

Particle beam ionization. :

Particle beam ionization.

Mass Analyser:-:

Mass Analyser:- They deflects ions down a curved tubes in a magnetic fields based on their kinetic energy determined by the mass, charge and velocity. The magnetic field is scanned to measure different ions. Types of mass analyzer:- (1) Quadrapole mass filter. (2) time of flight (3) Ion trap (4) Fourier transform ion cyclotron resonance (FT-ICR or FT-MS)

Ion trap:

Ion trap

Fourier transform ion cyclotron resonance (FT-ICR or FT-MS):

Fourier transform ion cyclotron resonance (FT-ICR or FT-MS)

Appplication of LC-MS:-:

Appplication of LC-MS:- Molecular weight determination Determining the molecular weight of green fluorescent proteins Structural determination e.g. structural determination of ginsenoside. Pharmaceutical application e.g. identification of bile acids metabolites. Biochemical application e.g. rapid protein identification using capillary lc/ms/ms.

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Food application e.g. identification of aflatoxin in food determination of vitamin D 3 in poultry feed supplement using MS 3 Environmental application e.g. detection of phenyl urea herbicides, detection of low level of carbaryl in food.


Wikimedia Foundation . 2010 . (1)   W. Paul & H. Steinwedel; Zeitschrift für Naturforschung , 8A; 1953 , p448. (2)   W. Paul; Agewandte Chemie - International Edition , 29; 1990 , p739. (3)   G. C. Stafford et al.; International Journal of Mass Spectrometry and Ion Processes , 60; 1984 , p85 and Analytical Chemistry , 59; 1987 , p1677. Referance:-

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