SAMPLING AND ANALYSIS OF VEGETABLE OIL SOURCE MATERIALS

AOCS Approved Procedure Am 5-04

Approved 2004

Rapid Determination of Oil/Fat Utilizing High

Temperature Solvent Extraction
DEFINITION
This method determines the substances extracted by petroleum ether under conditions of the test. The
compounds extracted, in most samples, are predominantly triacylglycerides. Small amounts of other
lipids along with minor components having some solubility in petroleum ether are also extracted.
SCOPE
This method is applicable to extractions of oil seeds (see Numbered Notes, 1), meats, feeds, and
foods and is equivalent to the petroleum ether extraction performed by the Butt and Soxhlet extrac-
tion apparatus.

APPARATUS 4mm of the top with the heat sealer encapsulating the
1. Analytical Balance, capable of weighing down to 0.1 sample.
mg. 4. Dry the samples in the bag at 100ºC for 3 hours (see
2. An extraction instrument capable of performing extrac- Numbered Notes, 4).
tions at elevated temperatures (90ºC-100°C) and pres- 5. Cool samples in a desiccator pouch to ambient temper-
sures (40-80 psi) on multiple samples. The apparatus ature and weigh (Wt. of pre-dried sample).
must also be capable of rinsing the sample and evapo- 6. Place up to 20 samples in the bag holder and extract in
rating the residual solvent (ANKOMXT10 Extractor or the instrument at temperatures between 90ºC-100ºC for
ANKOM XT20 Fat Analyzer, ANKOM Technology 15 to 60 minutes (see Numbered Notes, 5).
Corp.). 7. At the completion of the process remove the samples
3. Solvent and heat resistant filter bags, capable of being from the instrument and dry at 100ºC for 30 minutes.
heat sealed closed, retaining one micron particles and 8. Cool samples in a desiccator pouch to ambient temper-
permitting rapid solvent penetration (XT4 filter bags, ature and weigh (Wt. of extracted sample).
ANKOM).
4. Impulse heat sealer CALCULATIONS
5. Oven, capable of maintaining a temperature of 100ºC (Wt. of Pre-dried Sample
±2º. The oven should have sufficient convection flux − Wt. of Extracted Sample)
to clear the water vapor. % Oil/Fat = ––––––––––––––––––––––– × 100
6. Desiccator pouch (Moisture Stop weigh pouch, Wt. of Sample
ANKOM).
PRECISION
REAGENTS Results of the collaborative study in Table 1 indicates the
1. Petroleum ether—AOCS Specification H 2-41 (see precision (Sr, RSDr, r) that the analyst should use as a
Notes, Caution). benchmark for evaluation replication in the same laborato-
ry.
PREPARATION OF SAMPLE
1. Oilseed samples should be prepared according to NOTES
Section A of AOCS Methods using the methods speci- Caution
fied for each type of sample. Petroleum ether is extremely flammable. Avoid static elec-
2. Meat samples should be ground to a uniform consis- tricity. The explosive limits in air are 1-6%. A fume hood
tency with a food processor or similar device. should be used at all times when venting petroleum ether.
3. Feed samples should be ground to a particle size that
will pass through a 1-mm sieve and mixed thoroughly NUMBERED NOTES
(see Numbered Notes, 2). 1. Specific data on the accuracy and precision of soy-
4. Food samples should be processed with a food proces- beans, canola, safflower and corn are found in the data
sor or grinder to produce a representative sample of from the collaborative study (Table 1) and similar
uniform consistency. results have been found with other oilseeds extracted
with the Butt apparatus. Since this method does not
PROCEDURE regrind the sample after the original extraction and
1. Weigh filter bag then re-extract, for reference purposes AOCS Method
2. Weigh one gram (see Numbered Notes, 3) of prepared Am 2-93 is recommended.
sample directly in filter bag and avoid placing the sam- 2. If sample contains large amounts of H2O soluble com-
ple on the upper 4 mm of the bag (Wt. of Sample). ponents such as carbohydrates, urea, glycerol, and lac-
3. Completely seal the upper edge of the filter bag within tic acid remove by a water extraction. If H2O solubles

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Table 1
A summary of the statistical analysis of the international collaborative study of the Filter Bag Technique, coordinated by Andrew Komarek and Ronald Komarek
(ANKOM Technology Corp. Macedon, NY). Included in the summary is a comparison of the Filter Bag Technique with the results of analysis by AOCS certified
Laboratories using official AOCS or AOAC methods.
oat brownie soybean soybean corn poultry cattle pig cat bkft tortilla
Sample ID meal mix A canola meal A starter feed starter alfalfa food cookies cereal chips
Number of laboratories 12 12 11 9 12 12 11 10 11 11 12 11 12 12
Number of replicates 24 24 22 18 24 24 22 20 22 22 24 22 24 24
Collaborative Average, Oil/Fat % 5.8 8.7 20.9 39.0 1.6 3.3 3.3 3.2 5.6 2.4 6.3 22.7 2.3 19.9
Certified Labs Averagea 5.7 8.7 21.1 39.7 1.6 3.6 3.5 3.0 5.5 2.2 6.2 23.1 2.3 20.0
AOCS Am 2-93 Avg. for Oilseeds 21.9 44.8 3.6
repeatability
s(r) = repeatability std dev 0.36 0.20 0.35 0.23 0.14 0.31 0.24 0.18 0.20 0.39 0.27 0.20 0.26 0.39
Am 5-04 • Rapid Determination of Oil/Fat

RSD(r) = repeatability rel. std. dev 6.2 2.3 1.7 0.6 8.5 9.5 7.3 5.6 3.6 16.1 4.2 0.9 11.4 2.0
r = repeatability value 0.99 0.56 0.98 0.65 0.39 0.88 0.68 0.51 0.56 1.08 0.75 0.56 0.72 1.09
SAMPLING AND ANALYSIS OF VEGETABLE OIL SOURCE MATERIALS

Reproducibility
s(R) = reproducibility std dev 0.54 0.31 0.63 0.68 0.27 0.42 0.42 0.20 0.28 0.50 0.30 0.20 0.36 0.48
RSD(R) = reproducibility rel std. dev 9.4 3.5 3.0 1.7 16.3 12.7 12.6 6.1 5.0 20.7 4.7 0.9 15.7 2.4
R = reproducibility value 1.52 0.86 1.76 1.90 0.75 1.18 1.16 0.55 0.78 1.39 0.83 0.56 1.00 1.35
dog beef chicken soybean potato cheese corn
Sample ID food crackers turkey ham ground breast B safflower chips hot dog sausage corn B curls silage
Number of laboratories 12 10 12 9 11 11 11 9 11 12 11 12 12 12
Number of replicates 24 20 24 18 22 22 22 18 22 24 22 24 24 24
Collaborative Average 6.8 23.8 3.2 11.6 23.8 2.8 19.4 22.5 32.0 39.5 25.7 3.4 30.6 2.3
Certified Labs Averagea 6.9 24.0 3.2 11.3 23.5 2.7 19.7 23.0 32.0 39.0 25.0 3.7 30.5 2.3
AOCS Am 2-93 Avg. for Oilseeds 20.1 24.5
Repeatability (r)
s(r) = repeatability std dev 0.35 0.23 0.21 0.30 0.24 0.33 0.38 0.53 0.48 0.35 0.34 0.39 0.48 0.23
RSD(r) = repeatability rel. std. dev 5.23 0.96 6.57 2.59 1.01 11.89 1.97 2.36 1.49 0.89 1.33 11.48 1.59 9.87
r = repeatability value 0.99 0.64 0.58 0.84 0.67 0.94 1.07 1.49 1.34 0.98 0.96 1.10 1.36 0.63
Reproducibility (R)
s(R) = reproducibility std dev 0.35 0.23 0.34 0.30 0.36 0.33 0.62 0.83 0.52 0.59 0.51 0.41 0.69 0.51
RSD(R) = reproducibility rel std. dev 5.23 0.96 10.84 2.59 1.49 11.89 3.19 3.69 1.61 1.49 1.98 11.93 2.27 22.45
R = reproducibility value 0.99 0.64 0.96 0.84 0.99 0.94 1.73 2.33 1.45 1.65 1.43 1.14 1.94 1.44
aAOCS Official Methods Ba 3-38, AOAC 920.39 or equivalent
 SAMPLING AND ANALYSIS OF VEGETABLE OIL SOURCE MATERIALS
Am 5-04 • Rapid Determination of Oil/Fat

are present, weigh sample onto filter paper, extract

with five 20 mL portions of H2O allowing each portion
to drain. Place the filter paper containing the washed
sample into the filter bag and proceed with pre-drying
step.
3. A range of sample size from about 0.25 g to 3 g can be
analyzed with this method depending on the limitation
of sample availability or the need for larger samples
due to the lack of sample homogeneity.
4. Two hours or less should be used for samples that are
highly susceptible to oxidation.
5. Extraction times will be dependent on the type of sam-
ple and the fineness of sample grind. Samples com-
posed of animal tissue will require only 15 to 30 min-
utes. Samples containing plant tissue can range from
15 to 60 minutes.

REFERENCES

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