1. Characterization and Derivation of Glucose using PMP Spring 2013 Instrumental Real World Naomi Bryner David Millard

2. Background  Glucose, most common sugar and essential for life.  Simple sugar and can lead to further knowledge of more complex sugars.  Critical in the production of adenosine triphosphate (ATP).  ATP is the energy source used throughout the body.  Product analyzed with nuclear magnetic resonance (NMR) and capillary electrophoresis (CE) and infrared spectroscopy (IR).

3. Method - Derivatization  Created stock solutions of NaOH (0.3 M), PMP in MeOH (0.5 M), HCl (0.3 M) and a sodium borate buffer (0.2 M, 9.5 pH).  Dextrose (1.3512 g) added to RBF with NaOH (30 mL) and PMP (30 mL), heated in mineral oil bath with condenser at ~70°C for 30 min.  HCl (30 mL) added after cooling to neutralize reaction.

4. Method – Extraction, Isolation & Analysis  Sample and ethyl acetate (190 mL) added to separatory funnel for extract (x 3).  Aqueous layer evaporated to dryness  Rotory evaporator for ethyl acetate, methanol  Lyophilizer for water  After sample was dried, water was added to dissolve sample, followed by another round of evaporation to dryness.  Product was analyzed with NMR, CE, and IR.

5. Data - CE PMP peakProduct peak

6. Data - IR

7. Data - Proton NMR Theoretical Spectrum Experimental Spectrum

8. Data – 13 C NMR Theoretical Spectrum Resultant Spectrum

9. Results  Stock glucose can’t be examined directly; lack of chromophores.  13 C NMR: noisy baseline led to an increase of scans from 1024 to 7500 (overnight).  CE was comparison of PMP to product, NMR and IR were characterization.  Technological advancements caused difficulty in method replication (2002 -> 2013).  Week-long derivatization and tedious product isolation was worthwhile.  Ketone, permanent aspect of PMP in MeOH, was not detected in IR analysis, indicative of a product without excess PMP.

10. References  Bruice, P. Y. In Organic Chemistry; Folchetti, N., Mullaney, R., Kaveney, D., Eds.; Pearson prentice Hall: Upper Saddle River, NJ, 2007, 5 th Edition, pp 418-460.  Thibault, Pierre, and Susumu Honda. Capillary electrophoresis of carbohydrates. Totowa, NJ: Humana, 2002  Special Thanks to:  Dr. Foy, Dr. Hu, Dr. Halligan, Amanda MacPhearson, Jesse Diehl, Delaney Caudill