UPPSALA DIETARY FIBER METHOD (Theander, 1995)
1. DM/OM
Samples K,L
Dried & Ground (500mg)
MO/MS DETERMINATION
(oven / scale)
2. TOTAL FIBER
Samples A, B
ESTER PHENOLICS
Samples C, D
TOTAL PHENOLICS
Samples E, F
IVTF 24H
Samples G, H
IVTF 96H
Samples I, J
Dried & Ground (100mg)
Dried & Ground (200mg)
McDougall’s Buffer
Acetate Buffer
Ethanol (80%)
A.I.R. PREP
Enzymes
Acetone (1x)
(Alcohol Insoluble
Ethanol (95%)
Drying (N2)
Residue)
Water bath (96/24h_39ºC)
Frozen
Lyophilized
Rumen liquid (microbes)
TOTAL FIBER
A, B
IVTF 24h
G, H
H2SO4 (72%)
IVTF 96h
I, J
INOSITOL
DIw
AC
Filtration
Wash (90ºC)
UA COLORIMETRIC DETERMINATION
Glucuronic acid (std)
DERIVATIZATION CHO
NH4OH (12M)
DERIVATIZATION CHO
Glacial Acetic Ac.
(GC)
KLASON LIGNIN GRAVIMETRIC
DETERMINATION (OM: Oven/Scale)
1-Methylimidazole
Anhydride acetic
Ethanol (100%)
KBH4
KOH (2x)
EXTRACTION ETHYL ACETATE
(Microtubes w/ Na2SO4)
Evaporation Vial GC
GC Optima Grade Chloroform
GC VIAL
(N2)
a. ESTER PHENOLICS
C, D (2N)
TOTAL PHENOLICS
E, F (4N)
NaOH (2N) Water bath 39ºC (24h)
Steel bombs
NaOH (4N) Sand bath 160ºC (2h)
H3PO4
SPE
HPLC
WASH
VIAL
[NITROBENZENE OXIDATION (NBO)]