AOAC Official Method 934.

01

Moisture in Animal Feed

Loss on Drying at 95-100 C -Final Action

Dry amount of sample containing 2 g dry material to constant weight at 95-100 C under pressure
<100 mm Hg. For feeds with high molasses content, use temperature < 70 and pressure <50 mm.
Use covered Al dish >50 mm diameter and 40mm deep. Report loss in weight as moisture.

AOAC Official Method 925.04

Moisture in Animal Feed

by distillation with Toluene

A. Apparatus

Connect 250 mL flask of Pyrex or other resistant glass by means of Bidwell-Sterling moisture
receiver to 500 mm Liebig condenser. Calibrate receiver, 5 mL capacity, by distilling known
amounts water into graduated column, and estimating column of water to .01 mL. Clean tube and
condenser with chromic acid cleaning mixture, rinse thoroughly with water, then alcohol, and
dry in oven to prevent undue amount water from adhering to inner surfaces.

B. Determination

If sample is likely to bump, add dry sand to cover bottom of flask. Add enough toluene to cover
sample completely. Weigh and introduce enough sample into toluene to give 2-5 mL water and
connect apparatus. Fill receiving tube with toluene, pouring it through top of condenser. Bring to
boil and distill slowly, 2 drops/sec, until most of water passes over, then increase rate of
distillation to 4 drops/sec.

When all water is over, wash down condenser by pouring toluene in at the top for a short time to
see whether any more water distills over. If any water remains in condenser remove by brushing
down with tube brush attached to copper wire and saturated with toluene, washing down
condenser at the same time. Let receiving tube come to room temperature. If any drops adhere to
sides of tube, force them down, using copper wire with end wrapped with rubber band. Then
read volume of water and calculate to percent.
 AOAC Official Method 991.02

Moisture in Soft-Moist and Semi-Moist

Pet Foods

Karl Fischer Method First Action 1991First Action 1995

(Applicable to samples containing 20-30% moisture and other volatile materials.)

Method Performance:

Sr = 0.3; SR = 0.4; RSDr = 1.1%; RSDR = 1.7%

A. Principle

Water is extracted with methanol from pet food that contains other volatile components, and
aliquot is titrated with Karl Fischer reagent.

B. Apparatus and Reagents

(a) Karl Fischer titration assembly.- Manual or automatic, with stirrer.

(b) Karl Fischer reagent.- Stabilized, single solution (Fisher Scientific Co., SK3, or equivalent).
(Caution: If solution contains pyridine, see Appendix B, safety note on pyridine.) To standardize
reagent, add 100 mg H2O from weighing pipet, or other suitable device, to 30-50 mL pretitrated
CH3OH, and titrate with Karl Fischer reagent. C = mg H2O/mL reagent.

(c) Methanol.- ACS reagent grade. 99.8% CH3OH. 0.1% H2O. 0.001% acetone.

C. Sample Preparation

Reduce sample to as fine condition as possible. Household-style blender may be used; shake
blender back and forth to move sample into blades.

D. Determination
Accurately weigh 8-10 g prepared sample into Erlenmeyer that contains magnetic stirring bar.
Add 200 mL CH3OH, and stopper flask. Stir magnetically 15 min. Let solids settle. Transfer 10
mL aliquot to titration vessel containing pretitrated methanol and titrate with Karl Fischer
reagent. Determine blank on 10 mL CH3OH as above and subtract from sample determination.

Water, % = [2 * (mL reagent * C)]/g sample

Reference: JAOAC 73, 399(1990).

Revised: March 1996

AOAC Official Method 926.12

Moisture and Volatile Matter

In Oils and Fats

Vacuum Oven Method

Final Action

Soften sample, if necessary, by gentle heat, taking care not to melt it. When soft enough, mix
thoroughly with effective mechanical mixer.

Weigh 5 +/- 0.2 g prepared sample into Al moisture dish ca 5 cm diameter and 2 cm deep with
tight-fit slip-over cover. Dry to constant weight in vacuum oven at uniform temperature 20-25
degrees above bp of H20 at working pressure, which should be </= 100 mm Hg (13.3 kPa). Cool
in efficient desiccator 30 min and weigh. Constant weight is attained when successive 1 h drying
periods show additional loss of </= 0.05%. Report % loss in weight as moisture and volatile
matter.

Reference: Ind. Eng. Chem. 18, 1347(1926). JAOAC 14, 247(1931); 15, 560(1932).

AOAC Official Method 984.20

 Moisture in Oils and Fats

Karl Fischer Method

First Action 1984

Final Action 1985

ISO/TC34/SC11/N99 - ACOC Method

(Applicable to oils and fats except for alkaline or oxidized samples.)

A. Apparatus and Reagents

(A) Karl Fischer titration assembly Manual or automated, with stirrer.

(B) Karl Fischer reagent Stabilized with H2) equivalent of ca 5 mg H20/mL reagent. Available
commercially or prepare as follows: Dissolve 133g I2 in 425 mL dry pyridine in dry glass
stoppered bottle. Add 425 mL dry ethylene glycol monomethyl ether. Cool to <4 degrees in ice
bath and bubble in 102-105 g So2. Mix well and let stand 12 h. Reagent is reasonably stable, but
restandardize for each series of determinations. Standardize daily with sodium tartrate 2H2O. 1
mg sodium tartrate 2H2O = 0.1566 mg H2O. Alternatively, standardize with weighed H2O in
methyl alcohol as follows: Transfer accurately weighed amount (50 mg) H2O to titration vessel
and titrate to electrometric end point. Calculate C = mg H2O / mL reagent.

(C) Karl Fischer reagent diluent 2 Methoxyethanol - pyridine (4+1).

(D) Sample solvent Anhydrous CHCL3 - methyl alcohol (1+1) or (2+1).

B. Determination

Weigh to nearest 0.01 g, 5-25 g prepared sample, containing </= mg H2O, into tritation vessel,
dissolve in anhydrous CHCl3 - methyl alcohol. Titrate with undiluted or diluted (1+1) Karl
Fischer reagent to electrometric end point. Carry out blank test using same amount of reagent,
diluent, and solvents. Subtract blank titer.

% H2O = (mL reagent x C) / (g sample x 10)

Note: Pyridine-free Karl Fischer reagents are available from laboratory reagent suppliers.
Reference: JAOAC 67, 299(1984)