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MCAT lab techniques

lab techniques OC, Biochem, DNA

liquid-liquid extraction shaken and separated like like (polar-polar eg); salt from base+acid in aqueous solution;WB&COOH=anionic salt; phenols w/ NaOH diluted; NaHCO3 no phenol yes COOH
principle of extraction salts are preferred for aqueous phase
thin layer chromatography polarity & capillary action; results UV light maybe & Rf = substance move/solvent final move
polar stationary phase of TLC solid-liquid with SiO2 absorbent; developing/running
Column chromatography SiO2 gel gravity filtration with polar reacting with gel/TLC plate; least polar comes out first
Ion exchange chromatography gravity filtration with ionic resins, or salts, where the + charged will flow through and replace the Na+ for example; what does not stick, neutral and - out first; pI value of importance; pI values above pH first to elute
High Performance Liquid Chromatography (HPLC) high pressure forces mobile through stationary = efficient speed, no gravity; chromatogram; silica gel
reverse phase HPLC nonpolar is silica gel binds nonpolar; mobile is polar
size exclusion chromatography porous polymer beads; small travel more distance end up last
affinity chromatography stationary phase solid resin, like with Ab; binding with Ab goes gravity bottom; no centrifuge using beads magenetized; commercialized specificity; N/C affinity tag
gas chromatography mobile gas phase & stationary liquid phase; volatility
distillation breaking bonds; making bonds; BP
Simple distillation impurities miniscule; BP
Fractional distillation column packed with wire/beads/cotton; vapor impurities at bottom; pure at top
spectroscopy, general ground state to excited state to ground =emission & record
Mass spec high vacuum ionization; magnetic field & flight path change; r=mv/qB; abundance y axis & M/e x axis; good for isotopes; Br/Cl
UV Visible spec transition metal d orbital & conjugation pi bonds; wavelength related to amt of conjugation; absorption and emission opposite red-green;orange-blue;vio-yell
IR spec wavelength 2.5 to 20um; excited vibration; wavenumber is 1/wavelength=v/c; E=hv; RCOR 1700; C=C 1650; OH 3600-3200; RCCR 2260-2100; CH 3300-2700; N low O
H NMR spec, general equivalent Hs
spin-spin splitting H NMR; Hs not same group interact; protons in field interact with molecule proton; n + 1 & n=neighboring Hs; if 2 neighboring Hs interfere then 3 splits; singlet for 0 doublet for 1, triplet, quartet, quintet, sextet
NMR integration area under curve related to number of Hs causing the spike
Created by: Rocks2278