Method References
It is the policy of the University of Missouri Agricultural Experiment Station Chemical Laboratories to declare the primary method references for all official methods that are employed for the major assays routinely performed and/or requested. This is not meant to be an exhaustive listing, especially for 'research' procedures where several methods may be available, depending on sample matrix.
- Amino Acid Analyses
- Proximates, Other Fiber Analyses
- Sugars and Carbohydrates
- Lipids, Fats and Oils
- Mineral Analyses
- Water and Wastewater Analyses
- Fertilizer and Limestone
- Mass Spectrometry
Standard Amino Acid Analyses
- Complete Amino Acid Profile (AAP)
- AOAC Official Method 982.30 E(a,b,c), chp. 45.3.05, 2006
- Amino Acid Profile without Tryptophan (AAP-T)
- AOAC Official Method 982.30 E(a,b), chp. 45.3.05, 2006.
- Cysteine, Methionine and Lysine +9 (CML+9)
- JAOAC 70:171-174, 1987.
- Standard Protein Hydrolysate (SPH)
- AOAC Official Method 982.30 E(a), chp. 45.3.05, 2006.
- Methionine, Lysine or Hydroxyproline (M, L or HP)
- AOAC Official Method 982.30 E(a), chp. 45.3.05, 2006, acid hydrolysis.
- Cysteine & Methionine (CM)
- AOAC Official Method 982.30 E(b), chp. 45.3.05, 2006; peformic acid oxidation/acid hydrolysis.
- Tryptophan (T)
- by Enzymatic hydrolysis - colorimetric determination.
- by Alkaline hydrolysis - AOAC Official Method 988.15 , chp. 45.4.04, 2006
- Free Supplemental Amino Acids (FM, FL or FML)
- Methionine or Lysine, JAOAC 65:798, 1982.
- Methionine and lysine, JAOAC 65:798, 1982.
- AOAC Official Method 999.13
- Free Amino Acid Profile - Foods/Feeds (FAAP)
- Available Lysine (AL)
- AOAC Official Method 975.44, chp. 45.4.03, 2006.
- Available Lysine and Complete Amino Acid Profile (AL+AAP)
- AOAC Official Method 975.44 ; 982.30 E(a,b,c), chp. 45.3.05, 2006.
- HMB (also known as MHA and others) - Amino Acid Analogue for Methionine
- R.R.Ontiveros, W.S.Shermer, and R.A.Berner, An HPLC method for the determination of 2-hydroxy-4-(methylthio)butanoic acid (HMB) in supplemental animal feeds, J.Agric. Food Chem., 35(5), 692, 1987.
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Physiological Analyses of Biological Fluids
- Physiological Analyses
- Free amino acids in Plasma, Urine,Cerebrospinal Fluid and other biological fluids.
- Beckman 121-M Application Notes. 121-M-TB-004A, March 1975; and 121 MB-TB-018, January, 1980. Deyl, Z., Hyanek, J. and Horakova, M. (1986) Profiling of amino acids in body fluids and tissues by means of liquid chromatography. J. Chromatog. 379, 177-250.; Fekkes, D. (1996) State of the art-of-the-art high-performance liquid chromatographic analysis of amino acids in physiological samples. J. Chromatog. B. Biomed. Appli. 682, 3-22.
Microanalysis of Amino Acids
- Amino Acid Profile (Tryptophan not included, inquire)
- Hyp, Asp, Thr, Ser, Glu, Pro, Lys, Ala, Cys, Val, Met, Ile, Tyr, Phe, Hyl, His, Lys, Arg
- Standard Protein and Peptide Hydrolysis
- Hyp, Asp, Thr, Ser, Glu, Pro, Lys, Ala, Cys, Val, Met, Ile, Tyr, Phe, Hyl, His, Lys, Arg
- Sample in stained/unstained polyacrylamide gel.
- Lyophilized sample or in solution.
- Sample on PVDF, preferred, nitrocellulose, Immobilon (or similar membrane), resin, etc.
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Proximate Analyses
- Total Nitrogen or Crude Protein (CP)
- Combustion Analysis (LECO) AOAC Official Method 990.03, 2006
- Kjeldahl, AOAC Official Method 984.13 (A-D), 2006
- Standard report of 'crude protein' utilizes the calculation: 6.25 X Nitrogen value
- Ash Determination (Ash)
- AOAC Official Method 942.05, 2006
- Crude Fat (CF)
- By Ether Extraction, AOAC Official Method 920.39 (A), 2006
- Acid Hydrolysis, baked goods & pet food, AOAC Official Method 954.02, 2006.
- Crude Fiber (CFiber)
- AOAC Official Method 978.10, 2006
- Moisture (M)
- AOAC Official Method 934.01, 2006, vacuum oven.
- Total Carbohydrates, crude 'by difference'
- Calculation, 100% - %(CP+ Ash + Crude Fat + M).
- Food Energy - methods of analysis and conversion factors. Report of a technical Workshop, Rome, 3-6 December 2002, Food and Agriculture Association of the United Nations, Room 2003. FAO Food and Nutrition Paper 77. FAO Corporate Document Repository
- Available Carbohydrates
- Calculation, 100% - %(CP+ Ash + Crude Fat + M + Crude Fiber).
- Gross Calories
- Calculation: Protein = 4 calories/gram, Carbohydrate = 4 calories/gram, Fat = 9 calories/gram.
- Merrill, A.L. and Watt, B.K. 1973. Energy Value of Foods...Basis and Derivation. Agriculture Handbook No. 74. U.S. Government Printing Office. Washington, DC. 105p (out-of-print article site). Also: Food Labeling - Requirements for FDA Regulated Products, by James L. Vetter, E.M. Meloan, Ed., AIB International, Manhattan, KS, 2007.
- Acid Detergent Fiber (ADF) AOAC Official Method 973.18 (A-D), 2006.
- Neutral Detergent Fiber (NDF)
- JAOAC 56, 1352-1356, 1973.
- Cellulose (Cel) AOAC Official Method 973.18 (A-D), 2006.
- Cellulose = Acid Detergent Fiber (ADF)- Acid Detergent Lignin (ADL).
- See J. Dairy Sci. 89:2168-2176(2006) as an excellent research paper on method.
- Hemicellulose Calculation: NDF - ADF.
- Nitrogen-free Extract Calculation: 100 - (Moisture + Ash + Protein +Fiber + Fat).
- "Feeds and Feeding, Handbook for the Student and Stockman", 21st Ed., by Frank B. Morrison, The Morrison Publishing Co., Ithaca, NY, 1950.
- Lignin AOAC Official Method 973.18 (A-D), 2006.
- Total Dietary Fiber (TDF) AOAC Official Method 985.29, 2006.
- Pepsin Digestibility of Feeds and Foods
- Protein Dispersibility Index (PDI) AACC Official Method 46-24, 2006.
- Nitrogen Solubility Index (NSI) AOCS Official Method Ba 11-65, 2006.
- Protein Solubility, KOH method In House Protocol.
- Non-Protein Nitrogen
- Data for Biochemical Research, 1986, Rex. M.C. Dawson, et al., Editors, 3rd Ed., Oxford Science Publications, pp. 549-551.
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Sugars and Carbohydrate Analyses
- Phytate, in Foods (hexaphosphorylinositol)
- AOAC Official Method 986.11, 2006 -- may be modified to utilize ICP to analyze phosphorus.
- JAOAC 74:32-36, 1991.
- Sucrose, Raffinose, Stachyose
- J. Chromatogr., 153:539(1978); Biochim. Biophys. Acta, 222:339(1970); Handbook of Chromatography, 1982, Gunter Zweig and Joseph Sherma, Eds. In Chief, Shirley C. Churms, Ed., CRC Press, Boca Raton, FL.
- Uronic acid (as glucuronic acid)
- Analytical Biochemistry 4, 330-334 (1962) (Colorimetric; Carbazole).
- Anal. Biochem. 57:268-273 (1974) (Colorimetric; Harmine alternative to carbazole)
- Glycerol (also referred to as glycerin)
- Am. J. Enol. Vitic. 21:4:205-212 (1970) (gas-liquid chromatography, trimethylsilyl derivative).
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Lipids, Fats and Oils
- Fatty Acid Profile - saturated, mono- and polyunsaturates *
- AOAC Official Method 996.06 [Analysis of methyl esters by Capillary GLC]
- AOCS Official Method Ce 2-66 [Preparation of Methyl Esters of Fatty Acids].
- AOAC Official Method 965.49 [Preparation of Methyl Esters of Fatty Acids].
- AOAC Official Method 969.33 [Oils and Fat, Boron Trifluoride method].
- Omega 3 Fatty Acid Profile *
- AOCS Official Method Ce 1b-89 [Marine Oil - Fatty Acid Composition by GLC]
- AOCS Official Method Ce 1d-91 [n–3 and n –6 Fatty Acids by Capillary GLC]
- Trans Fats - total * AOCS Official Method Ce 1f-96 [Capillary GLC]
- Total Fatty Acids (Crude) AOAC Official Method 996.06 & AOCS Official Method Ca 5b-71
- Free Fatty Acids AOCS Official Method Ca 5a-40
- Conjugated Linoleic Acids (CLA, reported as 'total')
- AOCS Official Method Ce 1h-05 (include c9,t11- and t10,c12- octadecadienoic acids)
- Fat by Acid Hydrolysis AOAC Official Method 954.02, 2006
- Folch Extraction for Total Lipids from Animal Tissues
- Folch, J. et al., J. Biol. Chem. 226:497, 1957. (chloroform : methanol : water, 8:4:3, v/v/v)
- Bligh, E.G. and Dyer, J., Can. J. Biochem. Physiol. 37:911, 1959. (Folch extraction, variable solvent volumes, larger scale preparations)
- Peroxide Value of Fats and Oils
- AOAC Official Method 965.33
- AOCS Official Method Cd 8-53
- 4-Hr. or 20-Hr. AOM Peroxide Value
- Iodine Value AOCS Recommended Practice Ja 14-91
- TBA (Thiobarbituric Acid) Rancidity
- AOCS Official Method Cd 19-90
- Color, FAC AOCS Official Method Cc 13a-43
- Cholesterol
- AOAC Official Method 994.10 [in Foods by GLC]
- AOAC Official Method 970.51 [in Vegetable Fat/Oils by GLC]
- Phospholipids, Total AOAC Official Method 15:949.07
- Choline, Total Glick, D., J. Biol. Chem. 156:643, 1944
- Total Carotenoids AOAC Official Method 938.04
- Xanthophylls and Carotene AOAC Official Method 970.64
- Antioxidants **
- Ethoxyquin, in eggs, tissue and fat.
- BHA, BHT, PG, DG, TBHQ (in fats and oils)
*methods require 'crude fat' analysis for feeds, foods, grains, etc., that are not fats and oils. ** estimated ranges of all supplements must be declared.
Other Lipid, Fat, Specific Fatty Acids and Oil Analyses
- Insoluble AOCS Official Method Ca 3a-46
- Unsaponifiable AOCS Official Method Ca 6a-40
- Moisture AOCS Official Method Ca 2c-25
- Saponification Value AOCS Official Method Cd 3-25
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Inorganic, Elemental, Mineral Analyses
- Inductivel Coupled Plasma - Optical Emission Spectoscopy (ICP-OES)
- AOAC Official Method 985.01(A, B, D) Metals and Other Elements in Plants and Pet Foods.
- AOAC Official Method 990.08 Metals in Solid Wastes ICP Spectroscopic Method.
- Flame Atomic Absorption Spectroscopy (FAAS)
- AOAC Official Method 968.08, Minerals in Animal Feed and Pet Food.
- AOAC Official Method 975.03B(b) Metals in Plants and Pet Foods Atomic Absorption Spectrophotometric Method (applicable to Ca, Cu, K, Mg, Mn, and Zn) nitric-perchloric wet ash sample preparation with hydrochloric acid sample matrix
- Flame Emission Photometry - sodium, potassium
- AOAC Official Method 956.01
- Sulfur, gravimetric AOAC Official Method 956.01, 2006.
- Boron, volumetric AOAC Official Method 958.03, 2006.
- Chloride, manual titration AOAC Official Method 9.15.01, 943.01, 2006.
- Iodine, volumetric AOAC Official Method 935.14, 2006.
- Silicon, gravimetric AOAC Official Method 920.08, 2006.
- Titanium in Feeds & dried intestinal / feces
- Journal of Animal Science (2004) 82:179-183.
- Chromium in Feeds and dried Feces AOAC Official Method 990.08, 2006, ICP method.
- Dry Ash: Official Method 857.08 D(a), 2006.
- Nitric/Perchloric Wet Ash Sample Preparation: Official Method 968.088D(b), 2006.
- Selenium AOAC Official Method 996.16(G): Selenium in Feeds and Premixes: ICP method, 2006.
- Ytterbium in Feeds and dried Feces J. Dairy Sci. 75:2176-2183 I1992)
- Phosphorus, gravimetric AOAC Official Method 966.01 (plant tissue), 946.06, 2006. (feeds)
- Other mineral analyses (please inquire)
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Fertilizer Analyses
- Nitrogen (Kjeldahl method)
- AOAC Official Method 978.02, See also 920.02A(a),(c),(f),(i),(j),(k); 970.02A (a),(b); 970.02B & 955.05C.
- Available Phosphorus AOAC Official Method 960.03B(a)(1) & 978.01 A to H
- Soluble Potassium AOAC Official Method 983.02B(a) or (b) & 983.02 A, C-J
- Moisture vacuum dessication, AOAC Official Method 965.08B
Secondary and Micronutrients
- Calcium, Magnesium, Zinc, Copper, Iron, Manganese, Sodium
- Silicates in plant tissue
- Sulfur AOAC Official Method 980.02
- Boron AOAC Official Method 982.01B(a); determination 982.01 A, C and D
- Chloride
Limestone Analyses
- Calcium Carbonate Equivalent (CCE) AOAC Official Method 955.01A-B
- Magnesium (Mg)
- CCE by Neutralization and Magnesium (Mg)
- CCE plus sieve analysis profile, ENM* per ton
- CCE, Magnesium and sieves profile, ENM and EMG** per ton
- Sieve analysis profile only
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