39.1.16
AOAC Official Method 992.15 (h) Alumina oxide pellets.
Crude Protein in Meat and Meat Products
Including Pet Foods
Combustion Method
First Action 1992
(Applicable to meat and meat products [including pet foods] with
10–20% crude protein.)
Results of the interlaboratory study supporting acceptance of the
method:
sr = 0.12–0.41; sR = 0.18–0.46; RSDr = 0.60–2.23%; RSDR =
1.32–3.35%
A. Apparatus
(a) Combustion instrument.—Suitable for measuring 1–5%
nitrogen (ca 5–30% protein) in meat and meat products, to ±0.15%
of theoretical nitrogen content of standard, and be repeatable with
standard deviation of 0.15 for 10 successive determinations on same
standard. Instrument capable of analyzing single test portion of
³200 mg (to reduce impact of nonhomogeneity of meat and meat
products). Instrument to include oven to operate at ³850°C in O2
(for complete release of nitrogen from test portions), to isolate
nitrogen from other combustion products (i.e., CO2, H2O) for
subsequent quantitation, and capable of thermal conductivity
measurement of nitrogen (Leco FP428, Leco Corp., 3000 Lakeview
Ave, St. Joseph, MI, USA; Macro-N Analyzer, Foss Hereaus
Analysensysteme GmbH, Hanau 1, Ger many PE2410; or
Perkin-Elmer Corp., Norwalk, CT, USA, are suitable). Calibrations,
required by most instruments, must be conducted by using
theoretical % nitrogen in pure primary standard organic compounds,
such as EDTA.
(b) Food chopper.—With 18 in. (3 mm) plate, capable of grinding
meat test samples.
B. Reagents
The following reagents are typical but may vary depending on
instrument. Consult manufacturer’s instructions for specific
instruments.
(a) Compressed oxygen gas.—99.99%.
(b) Compressed helium gas.—99.99%.
(c) Compressed carbon dioxide.—99.99%.
(d) Compressed inert gas.—Nitrogen, or equivalent, oil and
water free.
(e) Ni tro gen stan dard.—Ethylenediaminetetraacetic acid
(EDTA), 9.59% nitrogen, or other suitable organic material of
high purity and known nitrogen content (e.g., nic otinic acid,
lysine hy drochloride).
(f) Quartz wool.
(g) Glass wool.
(i) Anhydrous magnesium perchlorate (MgClO4).
(j) Sodium hydroxide on silicate carrier.
(k) Silver tungstate on magnesium dioxide.
(l) Copper sticks.
(m) Copper metal turnings.
(n) Aluminum foil combustion cups.
(o) Silver vanadate.
(p) Copper oxide.
(q) Sn combustion cups or capsules.
Reagents available from several commercial manufacturers of
combustion analyzer instruments [see B(a)].
C. Preparation of Test Sample
Pass test samples through grinder 2´ in succession for emulsified
meat products, mixing thoroughly after each grinding; 3´ in
succession for nonemulsified (coarse or whole muscle) products.
D. Determination
Set instrument operating parameters (oven temperature, oxygen
flow, calibration values, etc.) according to manufacturer’s instructions.
Allow furnace and instrument to reach operating temperature and to
stabilize. Warmup time may be ca 6 h from cold start. Establish system
blanks as appropriate for analysis and calibrate blanks, if necessary. At
least 5 blank analyses are recommended. Calibrate instrument by using
3–5 analyses of nitrogen standard as follows:
(a) Accurately weigh 50–150 mg EDTA to the nearest 0.1 mg, or
equivalent amount of nitrogen if using other nitrogen standard, into
tared combustion cup or foil and transfer cup/foil to open loading port
on instrument. Enter or record protein conversion factor required, if
appropriate for instrument used.
(b) Close port, move boat into furnace, and begin analysis.
(c) When analysis is complete (3–5 min), repeat sequence for
next test portion.
(d) Adjust instrument as necessary based on results from
nitrogen standard.
(e) Analyze test portions by repeating steps (a)–(c).
(f) Read nitrogen results directly from instrument.
F. Calculations
Crude protein, % = nitrogen, % ´ 6.25
[Note: Results using this method average 1.01´ results using
928.08 (see 39.1.15).]
Reference: J. AOAC Int. 76, 787(1993).
Revised: March 1996
ã 2006 AOAC INTERNATIONAL