ANKOM Daisy - Manual (Page 13)

Appendix D
13
ANKOM Technology Method 3
In Vitro True Digestibility using the DAISY
P
II
P
Incubator
ANKOM
T e c h n o l o g y - 08/05
A. Reagents
(a) UBuffer Solution A:U
U
g/literU
KHB
2 B
POB
4 B
10.0
MgSOB
4 B
•7HB
2 B
O 0.5
NaCl 0.5
CaClB
2 B
•2HB
2 B
O 0.1
Urea (reagent grade) 0.5
(b) UBuffer Solution B:
NaB
2 B
COB
3 B
15.0
NaB
2 B
S•9HB
2 B
O 1.0
(c) Neutral Detergent Solution
B. Apparatus
(a) DAISYP II
P Incubator
(b) Filtration device - F57 Filter Bags
(c) Impulse bag sealer -
1915/1920 Heat Sealer.
(d) Thermos
(e) ANKOMP
200/220
P Fiber Analyzer
C. Procedure
U
Preparation of Filter Bags and Sample:
Pre-rinse F57 filter bags in acetone for three to five minutes and completely air-dry. The acetone
rinse removes a surfactant that inhibits microbial digestion. Weigh each F57 filter bag and record
weight (WB
1 B
). Zero the balance and weigh 0.25g of sample (WB
2 B
) directly into filter bag. NOTE: For
48 hr studies a sample size of 0.5 g is acceptable. Heat seal bag closed and place in the DaisyP II
P In-
cubator digestion jar (up to 25 samples per jar). Samples should be evenly distributed on both sides
of the digestion jar divider. Include at least one sealed blank bag for correction factor (CB
1 B
).
U
Preparation of (combined) Buffer Solution:U (For each digestion jar)
(a) Pre-warm at 39°C both buffer solutions (A & B). In separate container add ~266 ml of solu-
tion B to 1330 ml of solution A (1:5 ratio). The exact amount of A to B should be adjusted
to obtain a final pH of 6.8 at 39°C. No further adjustment of pH is necessary. Add 1600 ml
of combined A/B mixture to each digestion jar.
(b) Place the digestion jars with samples and buffer solution into DaisyP II
P Incubator and turn on
heat and agitation switches. Allow temperature of digestion jars to equilibrate for at least
twenty to thirty minutes.