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CHEM 355

CHEM 355. ADVANCED BIOCHEM. PLANT PIGMENTS. GO TO STATION WITH SAFETY SHEET WORK IN PAIRS TODAY. 2-4 LEAVES -viable/not dryed. Mortar& Pestle 0.5g sand & 15mL acetone. ISOLATE PIGMENT. MACERATE GRIND. BE QUICK ACETONE’S VERY VOLATILE. +anhydrous Na2SO4. MEASURE VOLUME. FILTER.

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CHEM 355

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  1. CHEM 355 ADVANCED BIOCHEM

  2. PLANT PIGMENTS • GO TO STATION WITH SAFETY SHEET • WORK IN PAIRS TODAY

  3. 2-4 LEAVES -viable/not dryed Mortar& Pestle 0.5g sand & 15mL acetone ISOLATE PIGMENT MACERATE GRIND BE QUICK ACETONE’S VERY VOLATILE +anhydrous Na2SO4 MEASURE VOLUME FILTER ZERO SPECTROMETER (acetone) FILL CUVETTE (GLASS) DILUTE IF NECESSARY (A>1) SCAN 400-700ηm & NEED ABSORBANCE λ =661.6,644.8,470 ηm FOR CALCULATION SCAN

  4. CONCENTRATE SAMPLE SEPARATE PIGMENTS & SCAN AGAIN APPLY EXTRACT TO PAPER DRAW & MEASURE Rf CHROMATOGRAPHY SOLVENT (9:1) PET ETHER:ACETONE CUT APART BANDS +TEST TUBES +acetone REPEAT IF NECESSARY CORK & LET SIT ~10MINS EXTRACT FROM PAPER INTO ACETONE STIR/SHAKE SCAN 400-700ηm RECORD ABSORBANCE MAX & λ DILUTE IF NECESSARY (A>1)

  5. ACID BASE PROPERTIES OF AMINO ACIDS • WORK IN PAIRS • PLOT AS YOU GO –(+ table) • VISUALIZE IT WITH A GRAPH! • WRITE DOWN SAMPLE INFO (ID , concentration, HCl, H2O)

  6. STANDARDIZE BASE WITH HCl CALCULATE BASE MOLARITY USE FOR CALCULATIONS CALIBRATE pH METER OBTAIN UNKNOWN AMINO ACID *NOTE SAMPLE ID & CONC Bring pH around 2 (1.8-2.3) Add 5 mL of HCl 0.1M (or more) CHECK pH BASE TITRATION PLOT AS YOU GO New

  7. VITAMIN A • WORK IN PAIRS • ENSURE SAPONIFICATION IS COMPLETE • PREPARE COLUMN – during saponification –watch for air bubbles -don’t let the column dry out • LOOK AT LABELS – what’s in the margarine (ingredients & concentration)

  8. OLIGONUCLEOTIDE SYNTHESIS • WORK IN PAIRS • Solvents – check labels • in fumehoods – watch your solvents! • λ = 504 ηm

  9. MILK PROTEINS – 2weeks • WEEK 1 -WORK IN PAIRS • CASEIN ISOLATION • PREPARE COLUMN FOR WHEY ISOLATION NEXT WEEK • SEAL WITH PARAFFIN • LEAVE ON BACK COUNTER WITH BEAKER BELOW

  10. MILK PROTEINS – 2weeks • WEEK 2 • WORK IN PAIRS • WHEY ISOLATION • CAREY or FIBRE OPTIC PROBE • TAKE ABSORBANCE READINGS AS YOU GO – DON’T WAIT TILL END

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