Chromatography

Terms: Mobile phase Stationary phase Eluent Eluate Volume flow rate #ml solvent/min travel in column Linear flow rate #cm column length/ 1 min Retention time Retention volume volume of mobile phase to elute the particular solute Dead time in GC, time to elute CH4 Adjusted retention time Relative retention/selectivity factor Retention factor/ capacity factor

Types of Columns: 1. Packed 2. Open tubular Types of Chromatography based on separation type 1. Adsorption solid sp, liquid/gas mp 2. Partition high boiling sp bonded to solid phase, gas mp, based on polarity a. Normal phase b. Reverse phase 3. Ion exchange a. Cation exchanger b. Anion exchanger 4. Molecular exclusion/ size exclusion a. Exclusion limit: MW>el, sple eluted immediately b. Permeation Limit: MW<PL: sple eluted very long We want EL<MW<PL to separate sample 5. Affinity Types of Chromatography based on 1. Liquid chromatography 2. Gas Chromatography 3. Supercritical Chromatography

Efficiency of Separation High diff in Tr, narrow peak width HETP/Plate Height

-meas of column efficiency -high N (theo plate), low HETP -proportinaliy bet variance of band and distance travelled Plate Height Equation/ Van Deemter equation A: eddy diffusion (pathway travelled) B: longitudinal A, B, C not equal to 0 : packed columns A equal to 0: open tubular Quantitative Parameter: Peak area, peak height concentration Qualitative: retention time Types of Chromatography based on 1. Liquid chromatography 2. Gas Chromatography 3. Supercritical Chromatography 1. Gas Chromatography - Isothermal and temperature programming ( for broad boiling range samples) Components: 1. Carrier gas He, Ar, N2, H2 2. flow controller or pressure regulator 3. injection port 4. column conventional packed and capillary column 5. detector Detector: 1. Flame ionization detector for hydrocarbons (not to H2O, CO2, SO2, CO, NOx) 2. Thermal conductivity detector -universal 3. Electron capture detector halogenated (pesticides) 4. Thermionic P and N 5. Flame photometric S and P

2. Liquid Chromatography - isocratic and gradient elution Components: 1. Eluent reservoir 2. Pump 3. Sample injector 4. Column 5. Detector Detector: 1. Refractive index changes in RI

2. UV/Vis based on absorption 3. Electrothermal voltam/amperometric based 3. Supercritical Chromatography -nonvolatile but large -linear pressure chromatography/density programming vs. isobaric mode High P, high density, high solvating power of SCF, low elution time Components: 1. Pump 2. Injector 3. Column 4. Detector (FID/ECD)

Potentiometry
Use electrodes to measure voltages that provide chemical info Cell components: 1. Reference electrode 2. Indicator electrode a. Salt bridge Ecell = Eindicator Ereference + Ej Junction potential develop across liquid junction at each end of salt bridge Reference electrodes: 1. Calomel r.e 2. Ag/AgCl r.e Kinds of indicator electrode: 1. Metallic indicator electrodes a. Electrode of first kind Ag/Ag+ b. Electrodes of second kind Ag/AgCl c. Inert electrode Pt, Au, Pd, C 2. Membrane electrodes/ Ion selective electrodes a. Glass electrode for pH i. Boundary potential develop at 2 surface of glass membrane b. Liquid membrane electrode potl across interface bet solution containing alayte and liquid ion exchanger; e.g Ca2+ c. Crystalline membrane electrodes inorganic crystals F- selective LaF3 doped with Eu2+ d. Solid state chemical sensors/ISFETS Applications of Potentiometry: 1. Direct Potentiometry (ISE) -TISAB (total ionic strength adjustment buffer) Quantitative: Nernst equation 2. Potentiometric Titration Quantitative: Volume of titrant at equivalence point

Electroanalytical
- current measurements 1. Electrogravimetric analysis o Controlled potential electrolysis with 3-electrode cell Metal electrode polarizable Reference electrode non polarizable 3 electrodes used: Working electrodes Reference electrodes Auxiliary electrodes E= Ecathode Eanode IR - overpotential Overpotential: to overcome Ea IR (ohmic potl): overcome electrical resistance of soln Concentration polarization: conc of electroactive species near electrode not same as in bulk soln 3ways to reach electrode surface: 1. diffusion conc gradient 2. convection stirring/boiling 3. migration-attraction/repulsion 2. Coulometry based on counting electrons a. Controlled Potential Coulometry b. Controlled Current Coulometry 3. Voltammetry 4. Polarography 5. Anodic Stripping Voltam 6. Pulse Polaograph 7. Amperometry o Measure electric current bet. A pair of electrodes that are driving on electrolysis reaction o Clark O2, blood glucose monitoring o Rotating Disk Electrode w.e Convection and diffusion control flux o Basis: limiting current/diffusion proportional to rate of diffusion 8. Biamperommetry