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Fiber

AACC International Method

32-20.01
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Insoluble Dietary Fiber
Final approval October 27, 1982; Reapproval November 3, 1999

Definition
To determine the neutral detergent-insoluble dietary fiber of food, food products, and feedstuffs.
Apparatus
1. Extraction apparatus with condenser to fit 600-ml tall-form lipless beaker
and hot plate adjustable to temperature that will bring 200 ml water from 25° to
rolling boil in 5–10 min.
2. Tall-form lipless beakers, 600-ml.
3. Coarse glass-frit (ASTM 40–60 µm) 60-ml filter funnels or crucibles.
4. Air oven maintained at 110–130°.
5. Air oven or incubator maintained at 37± 2°.
6. Desiccator with efficient desiccant.
7. Filtering device; suction flask with suitable holder for filtering funnel or
crucible.
8. Fine glass wool.
9. Lyophilizer (if necessary).
10. Centrifuge.
11. Grinding apparatus capable of reducing material to 20–30 mesh particles.
Reagents
1. Sodium lauryl sulfate, U.S. Pharmacopeia.
2. Disodium ethylenediaminetetraacetate dihydrate (disodium EDTA), reagent
grade.
3. Sodium tetraborate, decahydrate (borax), (Na2B4O7·10H2O), ACS grade.
4. Disodium phosphate, anhydrous, reagent grade.
5. Triethylene glycol.
6. Phosphoric acid, ACS grade.
7. Sodium sulfite, anhydrous, reagent grade.
8. Petroleum ether; boiling range 35–60°.
9. Sodium dihydrogen phosphate, anhydrous, reagent grade.
10. Acetone, laboratory grade.
11. α-Amylase.
12. Toluene, reagent grade.
Procedure
1. Prepare neutral detergent solution: Mix 18.61 g disodium EDTA and 6.81 g
Na2B4O7·10H2O with 150 ml water and heat until dissolved. Dissolve sodium
lauryl sulfate (30 g) and 10 ml 2-ethoxyethanol in 700 ml hot water and add to
first solution. Dissolve 4.56 g Na2HPO4 in 150 ml hot water and add to first
doi: 10.1094/AACCIntMethod-32-20.01

8. Air-dry at room temperature or lyophilize if necessary for grinding. cool to constant weight in desiccator. 2. P.1 with H3PO4 if necessary. Prepare samples by grinding to pass 20–30 mesh screen (1 mm). filter residue with suction. Dry at 110° at least 4 hr. 2. dry at 110° overnight. Amylase may be assayed for activity by the method of Sumner as described in Methods of Enzymology. corn. Agriculture Handbook 379.1M NaH2PO4. Rinse with about 75–80 ml acetone. prepare 0. 4.Fiber AACC International Method 32-20. and filter through coarse glass-frit filter. Reference Goering. shaped to form cup.. K. Dept. Note: If precipitate forms during storage. Weigh equivalent of 1.S. Centrifuge. Report as percent insoluble dietary fiber: (weight of filter + residue) − weight of filter × 100 weight of original sample Notes 1. 11. Add in following order: a) 100 ml neutral detergent solution (at room temperature).1M sodium phosphate buffer. eds. Extract three times with 25 ml petroleum ether per g of sample if fat content is >10%. O. α-Amylase should be chosen with care since some have been shown to exhibit residual hemicellulase activity. Add 1–3 g pad of fine glass wool. 1970. Weigh sample. Colowick and N. Add enough enzyme solution to filtered neutral-detergent fiber residue (from step 5) to more than cover sample (about 50 ml). decant. Allow about 10 ml to pass through to displace wash water. pea) and cereal grains (oats. soy. P. Remove plug. 7. 9. b) 0. Heat to boiling in 5–10 min. warm to 60° until precipitate dissolves. Hold at 37° for 18 hr (overnight). Plug bottom of filter and add several drops of toluene. With 0. and pour neutral-detergent fiber residue into center of glass wool while filtering with suction. Kaplan. S. and oats. Adjust pH to 6. This method has been applied to a number of food and feed systems including materials containing legumes (soy. It has been studied collaboratively using bran and flour from corn.01 Page 2 of 2 Insoluble Dietary Fiber (continued) solution.50 g Na2SO3.5% (w/v) α-amylase in 0. then reduce heat to maintain even boiling for 60 min. timed from onset of boiling. 149. tare. . to coarse glass frit filter.1M Na2HPO4 and 0. H. 6. 3. Suspend 2. Agric. 10. U. Forage Fiber Analyses. and Van Soest. Wash with at least 300 ml 100° water. wheat. and wash with at least 500 ml water. 1955. 5. p. and rye). J..0 g of original sample into beaker.9–7. wheat.1M phosphate buffer (pH 7). and cool in desiccator to constant weight (2–4 hr).