Characterization and
Derivation of Glucose using
PMP
Spring 2013 Instrumental Real World
Naomi Bryner
David Millard
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).
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.
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.
Data - CE
PMP peak
Product peak
Data - IR
Data - Proton NMR
Theoretical
Spectrum
Experimental
Spectrum
Data –
Theoretical
Spectrum
Resultant
Spectrum
13C
NMR
Results
 Stock glucose can’t be examined directly; lack of
chromophores.

13C
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.
References
 Bruice, P. Y. In Organic Chemistry; Folchetti, N.,
Mullaney, R., Kaveney, D., Eds.; Pearson prentice Hall:
Upper Saddle River, NJ, 2007, 5th 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