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Objective
Total dietary fiber (TDF) is determined by gelatinizing duplicate samples
(previously fat-extracted if fat content is >10%) with heat-stable -amylase,
digesting with protease and amyloglucosidase to remove protein and starch, and
diluting the aqueous digest with four volumes ethanol to precipitate soluble
dietary fiber. The residue is filtered; washed with 78% ethanol, 95% ethanol,
and acetone; dried; and weighed. One duplicate is analyzed for protein, the other
incinerated at 525 to determine ash. The TDF collected is corrected for method
blanks, which include protein and ash determinations. This method is applicable
to cereal grains and cereal-based products for nutritional labeling purposes.
Apparatus
1. Water baths. One for boiling water or steam and the other set at 60 and
equipped with either multistation shaker or multistation magnetic stirrer to
provide constant agitation of digestion beakers during enzymatic hydrolysis.
2. Fritted crucibles. Two per sample assayed. Porosity No. 2 (Pyrex No.
32940, coarse ASTM 4060 m; or Corning No. 36060 Bchner, fritted disk,
Pyrex 60 ml, ASTM 4060 m). Clean thoroughly. Heat 8 hr at 525, cool and
soak, then rinse in distilled water. Add about 0.5 g Celite to air-dried crucibles
and dry at 130 to constant weight (usually 1 hr). Cool and store in desiccator
until used.
3. Vacuum source. Vacuum pump or water aspirator equipped with in-line
double vacuum flask to prevent contamination of residue in case of water
backup.
4. Vacuum oven, set at 70. Alternatively, 105 air oven can be used.
5. Desiccator.
6. Muffle furnace.
7. Beakers, tall-form, 400-ml.
8. Balance, analytical, capable of weighing to 0.1 mg.
9. pH meter, calibrated to accuracy of 0.1 pH unit, using pH 4.0 and 7.0
buffers.
Reagents
1. Ethanol, 95% v/v, technical grade.
2. Ethanol, 78%. Place 207 ml water into 1-liter volumetric flask. Dilute to
volume with 95% ethanol. Mix well. Prepare more, if necessary.
3. Acetone, reagent grade.
4. Phosphate buffer, 0.08M, pH 6.0. Dissolve 1.400 g Na phosphate
anhydrous (Na2HPO4) or 1.753 g dihydrate and 9.68 g Na phosphate monobasic
monohydrate (NaH2PO4H2O) or 10.94 g dihydrate in about 700 ml distilled
water. Dilute to 1 liter with water. Check pH with pH meter.
doi: 10.1094/AACCIntMethod-32-05.01
Fiber
32-05.01
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Activity Tested
Citrus pectin
Stractan (larch gum)
Wheat starch
Corn starch
Casein
-Glucan (barley gum)
(Sigma, 7391)
Pectinase
Hemicellulase
Amylase
Amylase
Protease
0.1
0.1
1.0
1.0
0.3
95100
95100
01
02
02
-Glucanase
0.1
95100
Preparation of sample
Total dietary fiber should be determined on as-is basis on dried low-fat or
fat-free sample. Homogenize sample, weigh, and dry overnight in 70 vacuum
oven. Cool in desiccator, reweigh, and record weight loss due to drying. Drymill portion of dried sample to 0.30.5 mm mesh. If sample cannot be heated,
freeze-dry before milling. If high fat content (>10%) prevents proper milling,
defat with petroleum ether (three times with 25-ml portions per gram of
sample) before milling. When analyzing mixed diets, always extract fat
before determining TDF. Record weight loss due to fat. Correct final percent
dietary fiber determination for both moisture and fat removed. Store dry-milled
sample in capped jar in desiccator until analysis is run.
Method
Run blank through entire procedure along with samples to measure any
contribution from reagents to residue.
Fiber
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Fiber
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Correct final percent TDF for fat and/or water if defatting and/or drying of
sample was done during sample preparation step.
References
1. AOAC International. 1998. Official Methods of Analysis of AOAC International, 16th ed., 4th
rev. Method 985.29. The Association, Gaithersburg, MD.
2. Prosky, L., Asp, N. G., Furda, I., DeVries, J. W., Schweizer, T. F., and Harland, B. F. 1985.
Determination of total dietary fiber in foods and food products: Collaborative study. J. Assoc. Off.
Anal. Chem. 68:677.
3. Prosky, L., Asp, N. G., Schweizer, T. F., DeVries, J. W., and Furda, I. 1988. Determination of
insoluble, soluble, and total dietary fiber in foods and food products. J. Assoc. Off. Anal. Chem.
71:1017.