ANKOM TDF Dietary - Manual (Page 6)

Operator’s Manual
pg. 6
Rev E 12/2/14
Method Comparison (Manual vs. Automated)
AOAC 991.43 Method – IDF/SDF assay without automation
Below is the AOAC 991.43 method flow done manually by a technician using glass crucibles and beakers, vacuum to assist
with filtering, and water baths to control temperature and mixing. DE refers to Diatomaceous Earth.
Prepare for Test Digest Sample Isolate IDF Residue Isolate SDF Residue
1 Prepare crucibles 5 Heat to 95°C 17 Transfer to filter 26 Add 95% EtOH to fitrate
1a Ash crucibles overnight 6 Add buffer 18 Filter with vacuum assist 27 Precipitate for 60 min
1b Clean with vacuum 7 Add Amylase 19 Scratch filter bed as
needed 28 Transfer to filter
1c Soak 1 hr @ room temp 8 Digest at 95°C for 35 min 29 Filter with vacuum assist
1d Rinse with 3 solutions 9 Scrape beaker walls and
rinse with H
2
O to move
all residue to bottom
20 Rinse with 70°C H
2
O 30 Scratch filter bed as
needed
1e Prepare DE filter bed 21 Rinse with 78% EtOH
1f Dry @ 130°C 22 Rinse with 95% EtOH 31 Rinse with 78% EtOH
1g Cool 1 hr and weigh 10 Cool to 60°C 23 Rinse with Acetone 32 Rinse with 95% EtOH
2 Weigh samples 11 Add Protease 24 Dry IDF residue 33 Rinse with Acetone
3 Prepare H
2
O bath(s) 12 Digest at 60°C for 30 min 25 Weigh IDF residue 34 Dry SDF residue
4 Place samples in beakers 13 Add HCl 35 Weigh SDF residue
14 Add AMG
15 Check/adjust pH Calculate Results
16 Digest at 60°C for 30 min 36 Correct for Protein
37 Correct for Ash
38 Calculate results using
weights and corrections
AOAC 991.43 Method – IDF/SDF assay with ANKOM
TDF
Dietary Fiber Analyzer automation
Below is the AOAC 991.43 method flow when using the ANKOM
TDF
Dietary Fiber Analyzer. Steps highlighted in YELLOW
represent tasks done by a technician. Steps highlighted in GREEN (1a-1g, 9, 19, and 30) are no longer needed because of Filter
Bag Technology. All other steps (including mixing) are done automatically by the ANKOM
TDF Dietary Fiber Analyzer under
computer control. DE refers to Diatomaceous Earth.
Prepare for Test Digest Sample Isolate IDF Residue Isolate SDF Residue
5 Heat to 95°C 17 Transfer to filter 26 Add 95% EtOH to fitrate
6 Add buffer 18 Filter with pressure assist 27 Precipitate for 60 min
7 Add Amylase 28 Transfer to filter
8 Digest at 95°C for 35 min 29 Filter with pressure assist
20 Rinse with 70°C H
2
O
21 Rinse with 78% EtOH
22 Rinse with 95% EtOH 31 Rinse with 78% EtOH
1 Weigh filter bags 10 Cool to 60°C 23 Rinse with Acetone 32 Rinse with 95% EtOH
2 Weigh DE and samples 11 Add Protease 24 Dry IDF residue 33 Rinse with Acetone
3 Install filter bags 12 Digest at 60°C for 30 min 25 Weigh IDF residue 34 Dry SDF residue
4 Add DE & Sample to bags 13 Add HCl 35 Weigh SDF residue
14 Add AMG
15 Check/adjust pH Calculate Results
16 Digest at 60°C for 30 min 36 Correct for Protein
37 Correct for Ash
38 Calculate results using
weights and corrections
This section is just an overview of how the ANKOM TDF Dietary Fiber Analyzer
executes the AOAC 991.43 method. Please see the “IDF/SDF Analysis” and “TDF
Analysis” sections of this manual for detailed instructions on how to perform
IDF/SDF and TDF assays with the analyzer.