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Mass Spectrometry
-An old technique that has recently been exploited for organic analysis. -Particularly used for identification & structure determination. -Defined as a tool for the generation, measurement & interpretation of the positive ions based on their masses. Theory: -Produces positive ion spectra. -Not a band but a line spectra. -Upon bombardment of an organic compound with beam of electrons(energy10ev) M + e M+ +2e -It may lose an electron and yield a positive ion. -M+ is often unstable. -The imparted energy is distributed to various bonds. -If the accumulated energy sufficient to break the particular bonds, then -It (M +) undergoes fragmentation in specific manner. -The fragments produced are separated on the basis of their mass to charge ration, m/z. -Charge is usually unity, hence, m/z=m. -The separated fragments are recorded in proportion to their abundance. -A mass spectrum of the molecule is produced in the form of a plot of ion abundance vs. massto-charge ratio. Instrumentation:

i)Inlet system: -Introduces a very small amount of sample(a micromole or less) into spectrometer. -Contains means of volatilizing solids or liquid samples. ii)Ion source: -Converts the component of sample into ions. -Output is a stream of positive or negative ions(commonly positive). -Then accelerated into mass analyser. iv)Mass analyser: -Acts as grating in an optical spectrometer. -By applying magnetic field, ions are dispersed based on the mass-to-charge ratios of the analyte ions. - impinge upon the collector according to m/z = H2r2/2V Where, ( H=applied magnetic field, r= radious of ion path, V=acceleration voltage) -The radius of the ion path may be changed by varying either the magnetic field(H) or the accelerating voltage(V). v)Detector/amplifier -Convert beams of ions into an electrical signal. -Signals are intensified & sent to read out system. v) Collector/detector/amplifier -Convert beams of ions into an electrical signal. -Signals are intensified & sent to read out system. The ion-source, ion path & collector of the mass spectrometer must be under high vacuum (107mm Hg). Operation: -Vapour sample is introduced through inlet system. -Sp. is bombarded by electrons having an energy about 70 eV. -Positive ions are produced & accelerated between two plates. M+

Accelerating voltage Potential drop betewen two plates(1000-4000)V -Ions pass through the slit source. -On the way, deflected by a magnetic field(H) according to their m/z. -Exited through collector slit & impinge upon the collector. -The signal received is amplified & recorded. -The height or intensity of the resulting peak is proportional to the ion abundance. -The tallest peak is treated as 100% & other are expressed referring the peak.

Application: i)Qualitative a)Identification b)Molecular mass determination/structure elucidation. c)Detection of impurities ii)Quantitative

a)Identification: -Mass spectrum may be used as simply as confirmatory evidence of identity. -As a fingerprint , the mass spectra of the sample compound are compared with that of authentic sample. -The two mass spectra should be recorded under the identical condition. b)Molecular mass determination/structure -For structural elucidation, interpretation of the mass spectrum provides part of information required. -Other data of UV, IR, NMR usually be used in conjunction with MS. -In a line spectrum each line corresponds to a positive ion of specific mass. -In case of simply loss of one electron, the parent (P+) or mass(M+) corresponds to the exact molecular Wt. of the compound. -Isotopic composition of element in a molecule Method of determining element make-up of an ion or molecule is by a consideration isotope peaks. Isotope causes an additional peak in the mass spectrum of a compound. 1 H & 2H12 ,C & 13C, 16O & 18O. -Peak matching The molecular wt. of an may be determined to six decimal place comparing with known fluorocarbon peak. -Nitrogen rule An even molecular Wt. of a molecular ion(parent ion) reaquire s to contain no nitrogen or an even No. of nitrogen , odd molecular Wt. of a molecular ion requires an odd No. of nitrogen atoms. c)Detection of impurities: -Impurities can be detected up to ppm. level. If structures of the impurities are quite different from that of the main component.

-If molecular Wt. of contaminants more than that of main component. Other: -Amino acid sequence in peptides: Determination is possible by MS. -Drug metabolite study: Hydrazaline, chlorpromazine, pethidine & their metabolites.(LC/MS or GC/MS methods of choice) -Clinical, toxicological and forensic detection of trace drugs : Accidental or delibrate abuse or overdosage of drugs or toxic chemical can be detected Codeine in blood and urine. Morphine in bile & at the injection site(vein). Quinbarbitone & pentobarbitone in gastric content. (LC/MS or GC/MS methods of choice) ii)Quantitative -Currently , analysis is performed in combination with chromatographic or capillary electrophoretic column e.g. GC/MS, LC/MS etc. - Analyte concentrations are obtained directly from the heights of the mass spectral peaks. -Calibration plot of peak height vs. concentration is prepared & used for the analysis of unknown. Other techniques are: -Isotope dilution technique -selective ion monitoring technique