B - Chapter VI: Carbohydrates
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use (?)
serum, plasma or whole blood
whole blood values (?) lower than serum or plasma
10 - 15%
erum or plasma should be refrigerated and separated from the cells (?) to prevent substantial loss thru glycolysis
within1 hour
separated unhemolyzed serum at rm temperature is stable for
8 hr; ref temperature for 48 - 72 hr
preferred additive :
sodium fluoride (gray top)
: approximately 8 - 10 hr fast (not >16 hr)
FBS
: immediate processing to prevent bacterial utilization of glucose
CSF glucose
: only for monitoring of diabetics and not for diagnosis
urine sugar
Chemical methods − done in an
alkaline medium
− glucose + boiling water —-> (?) (enol anion form) : reduce metallic ions
eneidol
Alkaline Copper reduction Tests
Folin-Wu Nelson-Somogyi Neocuproine
total reducing substances (true reducing substances and saccharoids) measured
Folin-Wu
barium sulfate enables measurement of true reducing substances only
Nelson-Somogyi
Campbell and King method
Neocuproine
neocuproine (7,9- dimethyl-1,10-phenanthroline) —> cuprous-neocuproine complex (yellow-orange)
Neocuproine
arsenomolybdic acid —> arsenomolybdenum blue
Nelson-Somogyi
phosphomolybdic acid —> phosphomolybdenum blue
Nelson-Somogyi
Alkaline Ferric reduction tests
• Johnson method • Folin/ Prussian blue method • Hagedorn-Jensen method
− reverse colorimetric method − more specific than copper reduction
Alkaline Ferric reduction
− ferricyanide —> ferrocyanide
Alkaline Ferric reduction
Condensation methods
(a) with aromatic amines (b) with phenols
Ortho-toluidine method by Dubowski (reference method)
Condensation methods with aromatic amines
based on the ability of carbohydrates to form Schiff bases with aromatic amines
Condensation methods with aromatic amines
glucose + ortho-toluidine —-> N-glycosylamine: Schiff base (green) 630 nm
Condensation methods with aromatic amines