If you are using an HPLC to analyze the filtrate resulting from an assay in the
ANKOM
TDF
Dietary Fiber Analyzer, we recommend that you use a Sugar Pak
column in the HPLC to avoid issues associated with glycerol that might be
contained in the enzymes (depending on the enzymes that you are using).
General solutions common to AOAC 991.43, 985.29, and 2001.03 methods
(b) Ethanol 95%.
(c) Ethanol 78%—Place 821 ml of 95% ethanol into a 1 L volumetric flask. Dilute to volume with H
2
O.
(d) Diatomaceous earth (DE)—(ANKOM DE1, DE2, or equivalent).
(e) Acetone—reagent grade.
General solutions unique to the AOAC 991.43 method
(f) MES—2-(N-Morpholino)ethanesulfonic acid (No. M-8250, Sigma Chemical Co., or equivalent).
(g) TRIS—Tris(hydroxymethyl)aminomethane (No. T-1503, Sigma Chemical Co., or equivalent).
(h) MES-TRIS buffer solution—0.05M MES, 0.05M TRIS, pH 8.2 at 24°C. Dissolve 19.52 g MES and 12.2 g TRIS in 1.7
L H
2
O. Adjust pH to 8.2 with 6N NaOH, and dilute to 2 L with H
2
O. (Note: It is important to adjust pH to 8.2 at
24°C. However, if buffer temperature is 20°C, adjust pH to 8.3; if temperature is 28°C, adjust pH to 8.1. For
deviations between 20 and 28°C, adjust by interpolation.)
(i) Hydrochloric acid solution (AOAC 991.43)—0.561N. Dilute 93.5 ml of 6N HCl to 1 L with H
2
O.
General solutions unique to the AOAC 985.29 and 2001.03 methods
(j) Hydrochloric acid solution (AOAC 985.29)—0.65N. Dilute 325 ml of 1N HCl to 500 ml with H
2
O.
(k) Phosphate buffer solution—0.08M, pH 6.0. Dissolve 1.40 g Sodium Phosphate dibasic, anhydrous (Na
2
HPO
4
) (or 1.75
g dihydrate) and 9.68 g Sodium Phosphate monobasic monohydrate (NaH
2
PO
4
) (or 10.94 g dihydrate) in a 1 L
volumetric flask. Dilute to 1 L with H
2
O. Adjust pH to 6.0.
(l) Sodium Hydroxide Solution—0.55N. Dissolve 11.00 g Sodium Hydroxide (NaOH ACS) in a 500 ml volumetric flask.
Dilute to 500 ml with H
2
O.
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