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FOOD & FOOD SUPPLEMENTSEuropean Directory of Services

ACRYLAMIDE

PurposeApplicable for the determination of acrylamide in various foods.

Method facts Sample size: 5 g Limit of quantitation: 10 ppb Precision: Varies with matrix Method reference: United States Food and Drug Administration, Center

for Food Safety and Applied Nutrition Office of Plant & Dairy Foods and Beverages, Detection and Quantitation of Acrylamide in Foods (2002)

DescriptionAcrylamide is extracted from food samples with 0.1% formic acid. If product contains fat, petroleum ether is used prior to extraction with 0.1% formic acid. The extract is purified through a solid phase extraction (SPE cartridge). Acrylamide in the sample extract is determined using LC-MS/MS. The acrylamide is determined using least square linear regression with 13C3-labeled acrylamide as an internal standard. Ions monitored for acrylamide are m/z 55, 44, and 27 and for the internal standard m/z 58. The ratio of peak areas for m/z 55 (acrylamide) and m/z 58 (internal standard) are compared to those for standards over the standard curve range.

AFLATOXINS

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AMINO ACID PROFILE, FREE BY HPLC

PurposeApplicable to all samples for the following amino acids: L-aspartic acid, L-glutamic acid, L-serine, L-histidine, glycine, L-threonine, L-arginine, L-alanine, L-tyrosine, L-cystine, L-valine, L-methionine, L-phenylalanine, L-isoleucine, L-leucine, L-lysine, proline, L-asparagine, L-glutamine, L-citrulline, L-tryptophan and hydroxyproline.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.1 mg/g Precision: On a corn sample, examples of typical RSDs are as follows:

proline 4.8%, glutamic acid 5.0%, histidine 4.6%, aspartic acid 3.1%. Method reference: Henderson, J.W.; Ricker, R.D.; Bidlingmeyer, B.A.;

Woodward, C., Rapid, Accurate, Sensitive, and Reproducible HPLC Analysis of Amino Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA columns and the Agilent 1100 HPLC, Agilent Publication, 2000

R. Schuster, Determination of Amino Acids in Biological, Pharmaceutical, Plant and Food Samples by Automated Precolumn Derivitization and HPLC, J. Chromatogr., 1988, 431, 271-284

DescriptionThe sample is extracted in acid. Determination is by high-performance liquid chromatography (HPLC) with fluorescence or diode array detection. The primary amino acids are derivitized with fluorenylmethyl chloroformate before injection.

ASSAYS

Pricing available from a Covance representative. For updates to assays, refer to our online catalog at nutri.covance.com.1

2800-675-8375 nutri.covance.com

AMINO ACID PROFILE (HPLC), TOTAL

PurposeApplicable to all samples for the following amino acids: alanine, arginine, aspartic acid (including asparagine), cystine (including cysteine), glutamic acid (including glutamine), glycine, histidine, hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, tryptophan and valine.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.1 mg/g Precision: On an infant formula, examples of typical

RSDs are as follows: leucine 1.4%, aspartic acid 1.8%, tyrosine 2.5%, methionine 2.9%

Method reference: Henderson, J.W.; Ricker, R.D.; Bidlingmeyer, B.A.; Woodward, C., Rapid, Accurate, Sensitive, and Reproducible HPLC Analysis of Amino Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA columns and the Agilent 1100 HPLC, Agilent Publication, 2000

Barkholt and Jenson, Amino Acid Analysis: Determination of Cystine plus Half-Cystine in Proteins after Hydrochloric Acid Hydrolysis with a Disulfide Compound as Additive, Analytical Biochemistry, 1989, 177, 318-322

R. Schuster, Determination of Amino Acids in Biological, Pharmaceutical, Plant and Food Samples by Automated Precolumn Derivitization and HPLC, J. Chromatogr., 1988, 431, 271-284

DescriptionThe samples are hydrolyzed in 6N hydrochloric acid for 24 hours at approximately 110C. Phenol is added to the 6N hydrochloric acid to prevent halogenation of tyrosine. Cystine and cysteine are converted to S-2-carboxyethylthiocysteine by the addition of dithiodipropionic acid. Tryptophan is hydrolyzed from proteins by heating at approximately 110C in 4.2N sodium hydroxide.

The samples are analyzed by HPLC after pre-injection derivitization. The primary amino acids are derivitized with o-phthalaldehyde (OPA) and the secondary amino acids are derivitized with fluorenylmethyl chloroformate (FMOC) before injection.

ASH

PurposeApplicable for the determination of ash in most foods and feeds.

Method facts Sample size: 5 g Limit of quantitation: Most matrices - 0.1% Precision: On a dog food matrix, the RSD is 5.0% Method reference: AOAC 923.03

DescriptionOrganic matter is burned off by igniting the sample at 550C in an electric furnace. The remaining material is determined gravimetrically and referred to as ash.

ANISIDINE VALUE

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B12 BY HPLC

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B-VITAMIN PROFILE

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3 Pricing available from a Covance representative. For updates to assays, refer to our online catalog at nutri.covance.com.

CAROTENOIDS

Option 1: alpha Carotene, beta Carotene, LycopeneOption 2: Cryptoxanthin, Lutein, Zeaxanthin

PurposeApplicable to the determination for the following carotenoids in foods, feeds, premixes, pharmaceutical and nutrition supplements: lycopene, and -carotene, free lutein, lutein esters, -cryptoxanthin, or zeaxanthin. Vitamin A from carotenes may be calculated.

Method facts Sample size: 25 g for food or feed products; 2 g for

supplements and premixes Limit of quantitation: 0.0200 mcg/100 g, but can vary

by sample size Precision: Varies with matrix Method reference: Official Methods of Analysis of

AOAC INTERNATIONAL, Current Ed., Method 2005.07, AOAC INTERNATIONAL, Gaithersburg, MD, USA (modified)

L-CARNITINE

PurposeApplicable for the quantitation of L-carnitine ininfant formulas, powders and premixes.

Method facts Sample size: 25 g Limit of quantitation: 0.50 mg/100 g Precision: 4.15% on an infant formula control Method reference: Starey et al.; Journal of AOAC

INTERNATIONAL vol. 91, No. 1, 2008 (modified)

DescriptionThe sample is diluted in water and a deuterated analog of carnitine is added as an internal standard. It is then filtered and injected onto a Waters Symmetry C8, 50 mm x 2.1 mm, 3.5 m column, held in an oven set at 30.0C. Three mobile phases are used in a gradient consisting of 0.1% heptafluorobutyric acid (HFBA) in water, 0.1% HFBA in methanol and 80% water/20% methanol. The amount of L-carnitine is determined using HPLC-MS/MS.

B-VITAMINS BY HPLC-MS/MS

PurposeApplicable to the determination of up to seven B- vitamins in infant formula, various food and dietary supplement products. Vitamins included are thiamin(B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), pyridoxine (B6), biotin (B7) and folic acid (B9).

Method facts Sample size: 8 g Limit of quantitation:

Vitamin Common Name LOQ (g/g)

B1 Thiamin 0.05

B2 Riboflavin 0.025

B3 Niacin 0.375

B5 Pantothenic Acid 0.25

B6 Pyridoxine 0.05

B7 Biotin 0.025

B9 Folic Acid 0.025

Precision: Ranging around 2-5%, depending upon the vitamin

Method reference: Covance internal method

DescriptionSamples are extracted with an acidic aqueous-solventsolution followed by addition of a small amount ofalkaline solution to precipitate protein and aid withchromatography. Extracts are filtered and introduced to the HPLC-MS/MS system. Results are calculated againstknown reference standards. Stable Isotope internalstandards of each B-vitamin are included.

CAFFEINE

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FAT (ACID HYDROLYSIS)

PurposeApplicable for the determination of fat in most products.

Method facts Sample size: 2 g Limit of quantitation: Most matrices - 0.1% Precision: On a dog food matrix, the RSD is 1.55% Method reference: AOAC 922.06, 925.32,

933.05, 954.02

DescriptionThe sample is hydrolyzed with hydrochloric acid. The fat is extracted using ether and hexane. The extract is filtered through a sodium sulfate column. The solvent is evaporated from the remaining extract and the fat is dried and weighed.

Official Methods of Analysis of AOAC INTERNATIONAL, Current Ed., Methods 941.15, 2005.07, AOAC INTERNATIONAL, Gaithersburg, MD, USA (modified)

Quackenbush, F. W., Reverse Phase HPLC Separation of cis- and trans-Carotenoids and its Application to Beta Carotenes in Food Materials, Journal of Liquid Chromatography, 10: 643-653 (1987) (modified)

DescriptionLow fat samples are extracted with alcohol and/or tetrahydrofuran. Samples with a higher level of fat are saponified and extracted with hexane. Each sample is then injected on a reverse phase high performance liquid chromatography system (HPLC) with ultraviolet (UV) detection. Quantitation is achieved with a linear regression analysis.

CHLORIDE

PurposeApplicable for the determination of acid-soluble chloride in feeds, plants, food products and tissues.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 200 ppm Precision: On a cereal matrix, the RSD is 1.01% Method reference: AOAC 963.05, 969.10, 971.27

(modified)

DescriptionSamples are weighed, double-deionized water is added, and the solution is mixed thoroughly and made acidic with nitric acid. Chloride is determined potentiometrically by titrating with a silver nitrate solution to a predetermined endpoint.

CHOLESTEROL

PurposeApplicable for the determination of cholesterol in most matrices including foods, feeds and fecal samples.

Method facts Sample size: 5 g Limit of quantitation: 1 mg/100 g Precision: 2-5% depending upon matrix Method reference: AOAC 994.10

DescriptionThe sample is saponified using ethanolic potassium hydroxide. The unsaponifiable fraction that contains cholesterol and other sterols is extracted with toluene. The toluene is evaporated to dryness and the residue is dissolved in dimethylformamide (DMF). The samples are derivitized to form trimethylsilyl ethers. The derivitized cholesterol is quantitatively determined by gas chromatography using 5 a-cholestane as an internal standard.

CHOLINE

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FATTY ACID PROFILES

These tests are based on AOCS method Ce1b-89 with thefollowing options: Fatty acids C8-C22: Applicable to most food products Fatty acids C8-C24: Applicable to products containing

marine lipids EPA/DHA Fatty acids C4-C24: Applicable to products containing

dairy and those with EPA/DHA

PurposeThis procedure is used for traditional fatty acid profile analysis. The method detects fatty acids having 4 to 24 carbon atoms as methyl esters. The conditions specifiedin this method are not suitable for determining epoxy oroxidized fatty acids that have been polymerized.

Method facts Sample size: 2 g Limit of quantitation: Most matrices - 0.01% Precision: On a butter matrix, the RSDs for fatty acids

are 1-5% Method reference: AOCS methods Ce 1b-89, Ce 1-62,

Ce 1e-91, Ce 1e-07 and Ce 2-66

DescriptionThe extraction can vary depending upon the type of product being analyzed. For some products, the lipid is extracted, saponified and subsequently derivitized. For other products a direct saponification is performed followed by derivatization. The methyl esters of the fatty acids are analyzed by gas chromatography using external standards for quantitation.

FATTY ACID PROFILES (NLEA)

These tests are based on AOAC method 996.06 with thefollowing options: Fatty acids C8-C22: Applicable to most food products Fatty acids C8-C24: Applicable to products containing

marine lipids EPA/DHA Fatty acids C4-C24: Applicable to products containing

dairy and those with EPA/DHA

PurposeThis procedure is used for quantitation of NLEA total fat using fatty acid profile analysis to include trans fat, polyunsaturated fatty acids and monosaturated fatty acids. The method detects fatty acids having 4 to 24 carbon atoms as methyl esters. The conditions specified in this method are not suitable for determining epoxy or iodized fatty acids that have been polymerized.

Method facts Sample size: 2 g Limit of quantitation: Most matrices - 0.01% Precision: On a hydrogenated vegetable oil matrix, the

RSDs for fatty acids are 1-5% Method reference: AOAC 996.06, AOCS Ce 1j-07 and

Ce 1h-05

DescriptionThe extraction can vary depending upon the type of product being analyzed. For some products, the lipid is extracted, saponified and subsequently derivitized. Forother products, a direct saponification is performed followed by derivatization. The methyl esters of the fatty acids are analyzed by gas chromatography using external standards for quantitation.

FAT, TOTAL (ROESE-GOTTLIEB) BASE HYDROLYSIS

PurposeApplicable for the determination of fat in milk(liquid or powder), buttermilk, half and half, cream andice cream.

Method facts Sample size: 5 g Limit of quantitation: 0.1% Precision: 4.7% on a dry milk matrix Method reference: AOAC 989.05, 920.11, 932.06

DescriptionThe sample is hydrolyzed in a water bath usingconcentrated ammonium hydroxide. The fat is extracted using ether and hexane. The extract is evaporated, dried and weighed.

FIBER (McCLEARY)

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FIBER, TOTAL DIETARY (LEE)

PurposeApplicable to most foods.

Method facts Sample size: Solid matrices - 5 g/liquid matrices - 10 g Limit of quantitation: Most matrices - 1.0% Precision: On a cereal matrix, the RSD is 3.93% Method reference: AOAC 991.43

DescriptionDuplicate food samples are gelatinized with alpha-amylase and digested with enzymes in a Mes-Tris

buffer to break down starch and some protein. Solubledietary fiber in the aqueous enzyme digest is precipitated by treatment with ethanol. The digest is then filtered and the residue is washed with ethanol and acetone to remove starch, protein degradation products and moisture. The residue is dried and weighed. Protein content is determined for one of the duplicates; ash content is determined for the other. The total dietary fiber content of the sample is calculated after adjustment for the protein and ash values. Note: for products containing fructan sources, a fructan analysis may need to be performed.

FRUCTAN (HPLC) FOS

PurposeApplicable for the determination of fructans,which includes inulin and fructo-oligosaccharides (FOS). Well suited for high-level fructan samples.

Method facts Sample size: 20 g Limit of quantitation: Most matrices - 0.5% Precision: Varies with matrix Method reference: AOAC 997.08

DescriptionFructans are extracted from the matrix with water. The extract is centrifuged, filtered and an appropriate dilution is injected for the analysis of free fructose and sucrose. An aliquot of the filtrate is also subjected to treatmentby enzymes to liberate fructose from the fructans. The net fructose content is determined on an HPAEC using PAD and compared against known standards.

FRUCTAN (SPECTROPHOTOMETRIC) FOS

PurposeApplicable for the determination of fructans, which includes: fructo-oligosaccharides (FOS) and inulin.

Method facts Sample size: 5 g Limit of quantitation: Most matrices - 0.5% Precision: On a spiked flour matrix, the RSD is 2.04% Method reference: AOAC 999.03

DescriptionThe sample is extracted with water. The extract is then treated with enzymes to hydrolyze and remove sucrose,starch and maltosaccharides. An aliquot of the filtrate is also subjected to treatment by enzymes to liberate fructose and glucose from fructan. The amount of these reducing sugars from fructan is determined by spectrophotometric methods.

FREE FATTY ACIDS (TITRATION)

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GLYCEROL

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INORGANIC ANALYSIS BY ICP

PurposeApplicable for the determination of up to 20 elements (ppm levels) in virtually all matrices. See below for options. Options 3 and 4 are matrix dependent. Please call to verify if the package price is applicable to your sample.

Option 1: Calcium, iron, sodium (for nutrition labeling)Option 2: Calcium, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, zincOption 3: Option 2 plus aluminum, barium, boron, chromium, molybdenum, strontiumOption 4: Option 3 plus beryllium, cadmium, cobalt, nickel, vanadium

Individual elements can be performed on request.

Method facts Sample size: 10 g Limit of quantitation: Element and matrix dependent Precision: For most elements, the RSD is


IODINE (ICP-MS)

PurposeApplicable for the determination of total iodine in a variety of matrices including foods (i.e. powders, liquids, dairy products, infant formulas), nutritional supplements and mineral premixes at levels ranging from low ppm to percent.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.1 ppm Precision: On a milk powder matrix, the RSD is


MOISTURE, 100C (VACUUM OVEN)

PurposeApplicable for the determination of moisture in foods and feeds, except for fruit, fruit products, vegetables, vegetable products, sugar and sugar products.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.1%

Precision: On a dog food matrix, the RSD is 2.9% Method reference: AOAC 925.09, 926.08

DescriptionThe sample is dried in a vacuum oven at 100C to a constant weight.

MOISTURE (KARL FISCHER)

PurposeApplicable for the determination of water content in most matrices involving a USP or FCC monograph.


Precision: Varies with matrix Method reference: USP

DescriptionThe sample is dissolved or suspended in 50 mL of solvent and titrated with Karl Fischer moisture reagent. Milli-Q water is used to calibrate the instrument.

NUCLEOTIDES

PurposeApplicable for the determination of the nucleotidescytidine-5-monophosphate (5CMP), uridine-5-monophosphate (5UMP), guanosine-5-monophosphate (5GMP), inosine-5-monophosphate (5IMP) and adenosine-5-monophosphate (5AMP) in infant formula and some food products.

Method facts Sample size: 5 g Limit of quantitation: Most matrices - 3.0 mg/100 g Precision: On an infant formula sample, the RSDs are

as follows: 5AMP - 1.85%, 5IMP - 4.66%, 5GMP - 2.24%, 5CMP - 1.70%, 5UMP - 4.58%

Method reference: Paubert-Braquet M., Dupont CH. Paoletti R, Foods, Nutrition and Immunity, Quantification of Nucleotides in Human Milk and their effects on Cytokine Production by Murine Fibroblasts, J774A1 Macrophages and Human Monocytes, DYN Nutr Res

Basel, Karger, 1:22-34 (1992)

DescriptionThe nucleotides are extracted into an aqueous, protein-free filtrate; collected on a solid-phase extraction (SPE) strong anion exchange (SAX) column; and subsequently eluted with phosphate buffer. The eluted nucleotides are quantified using HPLC equipped with UV detection against a standard of known concentration.

NITRATE/NITRITE

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MOISTURE, 70C (VACUUM OVEN)

PurposeApplicable for the determination of moisture in fruits and vegetables.


Precision: On a ground carrot matrix, the RSD is 1.00% Method reference: AOAC 934.06

DescriptionThe sample is dried in a vacuum oven at 70C to a constant weight.


PEROXIDE VALUE

PurposeApplicable for the determination of peroxide value in all normal fats and oils and extracted fats and oils. This method is highly empirical and any variation in procedure may result in variation in results.

Method facts Sample size: 20 g of food/feeds, 5 g of oil Limit of quantitation: NA Precision: NA

Method reference: AOCS method Cd 8-53 AOAC extraction 983.23 USP fats and fixed oils

DescriptionThis method determines all substances, in terms of milli-equivalents of peroxide per kilogram of sample, that oxidize potassium iodide under the conditions of the test. These are assumed to be peroxides or other similar products of fat oxidation.

PESTICIDE MULTIRESIDUE ANALYSIS (300+ COMPOUNDS)

PurposeApplicable for the determination of pesticides in various food, dietary supplement, infant formula and other related matrices.

Method facts Sample size: 25 g (50 g for high-moisture samples,

such as fruits and vegetables) Limit of quantitation: 0.01 mg/kg for most analytes

and matrices Precision: Varies with matrix Method reference:

1AOAC Official Method 2007.01 2CEN Standard Method EN 15662 3 Anastassiades, M.; Lehotay, S.J.; Stajnbaher, D.; Schenck, F.J. Journal of the AOAC INTERNATIONAL, 2003, 86, 412-431

4 Lehotay, S.J.; Mastovska, K; Lightfield, A.R. Journal of the AOAC INTERNATIONAL, 2005, 88, 615-629

5 Lehotay, S.J.; Mastovska, K.; Yun, S. J. Journal of the AOAC INTERNATIONAL, 2005, 88, 630-638

6 Mastovska, K.; Dorweiler, K.; Lehotay, S.J.; Wegscheid, J.; Szpylka, K. J. Agric. Food Chem., 2010, 58, 5959-5972

DescriptionThe sample extraction and clean-up are based on the QuEChERS method.1-6 The final extract is split for the analysis of GC- and LC-amenable pesticides using gas chromatography-tandem mass spectrometry (GC-MS/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. If requested, the results can be corrected to potential matrix effects using the method of standard addition, which provides the most accurate quantification.

ORGANIC ACID PROFILES

PurposeApplicable for the analysis of organic acids in foodsand food products. Please refer to the following options:Option 1: Sorbic and benzoic acidOption 2: Citric acid, malic acid, lactic acid and acetic acid

Method facts: Option 1 Sample size: 2 g Limit of quantitation: Option 1: 5 ppm Precision: 5.11% Method reference: Journal of the AOAC

INTERNATIONAL, 70(5) (1987)

DescriptionSamples are extracted with water and methanol. The organic acids are separated using reverse-phase HPLC and measured with UV detection.

Method facts: Option 2 Sample size: 5 g Limit of quantitation: 0.05% Precision for Option 2: 3.65% Method reference: AOAC 986.13

DescriptionSamples are extracted with a weak H2SO4 solution. Organic acids are separated using reverse-phaseHPLC and measured with UV detection. Results arequantified by comparing with a known reference standard.


RESISTANT MALTODEXTRIN (FIBERSOL) WITH TOTAL DIETARY FIBER

PurposeApplicable for the determination of food products containing resistant maltodextrins.

Method facts Sample size: Powder samples 5 g, liquid samples 20 g Limit of quantitation: Most matrices - 1.0% Precision: On a cereal matrix, the RSD is 3.36% Method reference: AOAC 2001.03 (modified)

DescriptionDuplicate samples are gelatinized with alpha-amylase and digested with enzymes to break down starch and protein. Ethanol is added to each sample to precipitate the soluble fiber. The samples are filtered, and the filtrate is purified and then concentrated. The sample is then analyzed using an HPLC system with refractive index detection.

SELENIUM BY ICP-MS

PurposeApplicable for the determination of selenium in a variety of matrices including, but not limited to, infant and adult nutritional formulas, rice, wheat, soy and seeds (e.g. canola).

Method facts Sample size: 10 g Limit of quantitation: 0.030 ppm Precision: The RSD is 3.32% on a non-fat milk powder,

2.08% on a wheat flour, 1.83% on a rice flour, 2.39% on a powdered infant formula, 1.80% on an infant/adult nutritional formula, 7.76% on soy meal and 1.24% on canola meal

Method reference: Official Methods of Analysis of AOAC INTERNATIONAL 18th Ed., AOAC

INTERNATIONAL, Gaithersburg , MD, USA, Official First Action Method 2011.19 (2011) (modified)

DescriptionThe sample is digested with concentrated nitric acid and water using a closed-vessel microwave digestion system. After digestion, the sample is transferred to a single-use disposable plastic vessel, then brought to a final volume with water. To normalize the organic contribution between sample and standard, a dilution is prepared for analysis containing methanol. The amount of selenium is determined with inductively coupled plasmamass spectrometry (ICP-MS) by comparing the counts generated by the unkowns to those generated by standard solutions.

SORBITOL

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PROTEIN (DUMAS)

PurposeApplicable for the determination of protein in most foods and feeds.

Method facts Sample size: 1 g Limit of quantitation: Most matrices - 0.1% Precision: On a cereal matrix, the RSD is 0.9% Method reference: AOAC 968.06

DescriptionNitrogen is determined by a combustion-detection technique. Nitrogen is released at high temperatures into a pure oxygen atmosphere and measured by thermal conductivity. The percent nitrogen is converted to the protein using a factor of 6.25.


TAURINE

PurposeApplicable for the determination of taurine in most food, dietary supplements and feeds.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.015 mg/g Precision: On a cat food matrix, the RSD is 3.5% Method reference: Henderson, J.W.; Ricker, R.D.;

Bidlingmeyer, B.A.; Woodward, C., Rapid, Accurate, Sensitive, and Reproducible HPLC Analaysis of Amino Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA columns and the Agilent 1100 HPLC, Agilent Publication, 2000

R. Schuster, Determination of Amino Acids in Biological, Pharmaceutical, Plant and Food Samples by Automated Precolumn Derivitization and HPLC, Journal of Chromatography, 1988, 431, 271-284

AOAC 999.12

DescriptionThe sample is hydrolyzed in hydrochloric acid and is quantitated using an automated amino acid analyzer.

THIAMIN (VITAMIN B1)

PurposeApplicable for the determination of thiamin inmost foods, dietary supplements and feeds.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.01 mg/100 g Precision: On a commercial egg noodle sample, the

RSD is 2.44% Method reference: AOAC 942.23, 953.17, 957.17

DescriptionThe sample is autoclaved in dilute acid to extract the thiamin. The resulting solution is incubated with a buffered enzyme solution to release bound thiamin and hydrolyze starch. The solution is purified on an ion-exchange column. An aliquot is taken and reacted with potassium ferricyanide to convert thiamin to thiochrome. The thiochrome is extracted into isobutyl alcohol and read on a fluorometer against a known standard.

SUGAR PROFILE BY GC

PurposeApplicable for the determination of fructose, galactose, glucose, sucrose, lactose and maltose in a variety of matrices.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.1% Precision: On a cereal matrix, the RSDs for fructose,

glucose, sucrose and maltose are 2.22%, 1.99%, 1.19% and 6.01%, respectively

Method reference: Mason, B.S. and Slover, H.T., A Gas Chromatographic Method for the Determination of Sugars in Foods, Journal of Agricultural and Food Chemistry, 19(3):551-554 (1971)

Brosbt, K., Gas Liquid Chromatography of Trimethylsilyl Derivations, Methods in Carbohydrate Chemistry, Academic Press, New York, NY, 6:3-8 (1972)

DescriptionSugars are extracted from the sample with water and methanol. Aliquots are dried under inert gas and reconstituted with a hydroxylamine hydrochloride solution in pyridine containing phenyl--D-glucoside as the internal standard. The resulting oximes are converted to silyl derivatives with hexamethyldisilazane (HMDS) and trifluoracetic acid (TFA) treatment and analyzed by gas chromatography using a flame ionization detector.


USP 561 PESTICIDES

PurposeApplicable, but not limited, to dietary supplement products and raw materials for the analysis of pesticideslisted in the US and European Pharmacopeias (USP andEP).1,2

Method facts Sample size: 25 g Limit of quantitation: Varies by compound; USP and

EP have set limits Precision: Varies with matrix Method reference:

1 USP34-NF29:561 Articles of Botanical Origin. In: The United States Pharmacopeia, 2011

2 EP 07/2008:20813 Pesticide Residues. In: The European Pharmacopoeia, 7th Edition, 2011

3 AOAC Official Method 2007.01, Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning with Magnesium Sulfate

4 CEN Standard Method EN 15662: Food of plant origin Determination of pesticide residues using GC-MS and/or LC-MS/MS following acetonitrile extraction/partitioning and clean-up by dispersive SPE QuEChERS method

5 Anastassiades, M.; Lehotay, S.J.; Stajnbaher, D.; Schenck, F.J., Fast and easy multiresidue method employing acetonitrile extraction/partitioning and dispersive solid-phase extraction for the determination of pesticide residues in produce. Journal of AOAC INTERNATIONAL 2003, 86, 412-431

6 Lehotay, S.J.; Mastovska, K; Lightfield, A.R., Use of buffering and other means to improve results of problematic pesticides in a fast and easy method for residue analysis of fruits and vegetables. Journal of AOAC INTERNATIONAL 2005, 88, 615-629

7 Lehotay, S.J.; Mastovska, K.; Yun, S. J., Evaluation of two fast and easy methods for pesticide residue analysis in fatty food matrixes. Journal of AOAC INTERNATIONAL 2005, 88, 630-638

8 Mastovska, K.; Dorweiler, K.; Lehotay, S.J.; Wegscheid, J.; Szpylka, K., Pesticide multiresidue analysis in cereal grains using modified QuEChERS method combined with automated direct sample introduction GC-TOFMS and UPLC-MS/MS techniques. J. Agric. Food Chem., 2010, 58 5959-5972

9 Lehotay, S.J.; Ae Son, K.; Kwon, H.; Koesukwiwat, U.; Fu, W.; Mastovska, K.; Hoh, E.; Leepipatpiboon, N., Comparison of QuEChERS sample preparation methods for the analysis of pesticide residues in fruits and vegetables. J. Chromatogr. A 2010, 1217, 2548-2560

TOCOPHEROLS (TOTAL)

PurposeApplicable for the determination of alpha, beta, gamma and delta tocopherol in oils, foods, feeds and nutritional products.

Method facts Sample size: 10 g Limit of quantitation: Most matrices - 0.5 mg/100 g Precision: On a infant formula matrix, the RSD is

6.10%

Method reference: Cort, W.M.; Vicente, T.S. and Williams, B.D., Journal of Agricultural Food Chemistry, 31:1330-1333 (1983)

DescriptionThe product is extracted with an organic solventfollowed by separation on an HPLC silica column, andquantitation using fluorescence detection.

TRYPTOPHAN

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Pesticide

USP/EP Limit (mg/kg)

Acephate 0.1

Alachlor 0.05

Aldrin and dieldrin (sum of) 0.05

Azinphos-ethyl 0.1

Azinphos-methyl 1

Bromide, inorganic (calculated as bromide ion)*** 50

Bromophos-ethyl 0.05

Bromophos-methyl 0.05

Bromopropylate 3

Chlordane (sum of cis- and trans- isomers and oxychlordane)

0.05

Chlorfenvinphos 0.5

Chlorpyrifos-ethyl 0.2

Chlorpyrifos-methyl 0.1

Chlorthal-dimethyl 0.01

Cyfluthrin (sum of isomers) 0.1

Cyhalothrin, - 1

Cypermethrin (sum of isomers) 1

DDT (sum of o,p-DDT, p,p-DDT, o,p-DDE, p,p-DDE, o,p-DDD, and p,p-DDD)

1

Deltamethrin 0.5

Diazinon 0.5

Dichlofluanid 0.1

Dichlorvos 1

Dicofol 0.5

Dimethoate and omethoate (sum of) 0.1

Dithiocarbamates (expressed as CS2)*** 2

Endosulfan (sum of isomers and endosulfan sulfate) 3

Endrin 0.05

Ethion 2

Etrimphos 0.05

Fenchlorphos (sum of fenchlorphos and fenchlorphos-oxon)

0.1

Fenitrothion 0.5

Fenpropathrin 0.03

Fensulfothion (sum of fensulfothion, fensulfothion-oxon, fensulfothion-oxon sulfone and fensulfothion sulfone)

0.05

Fenthion (sum of fenthion, fenthion-oxon, fenthion-oxon sulfone, fenthion-oxon sulfoxide, fenthion sulfone and fenthion sulfoxide)

0.05

Fenvalerate 1.5

Pesticide

USP/EP Limit (mg/kg)

Flucytrinate 0.05

Fluvalinate, - 0.05

Fonofos 0.05

Heptachlor (sum of heptachlor and cis- and trans-heptachlor epoxides)

0.05

Hexachlorobenzene 0.1

Hexachlorocyclohexane isomers (other than ) 0.3

Lindane (-hexachlorocyclohexane) 0.6

Malathion and malaoxon (sum of) 1

Mecarbam 0.05

Methacriphos 0.05

Methamidophos 0.05

Methidathion 0.2

Methoxychlor 0.05

Mirex 0.01

Monocrotophos 0.1

Parathion-ethyl and paraoxon-ethyl (sum of) 0.5

Parathion-methyl and paraoxon-methyl (sum of) 0.2

Pendimethalin 0.1

Pentachloranisol 0.01

Permethrin (sum of isomers) 1

Phosalone 0.1

Phosmet 0.05

Piperonyl butoxide 3

Pirimiphos-ethyl 0.05

Pirimiphos-methyl (sum of pirimiphos-methyl and N-desethyl-pirimiphos-methyl)

4

Procymidone 0.1

Profenophos 0.1

Prothiophos 0.05

Pyrethrum (sum of cinerin I, cinerin II, jasmolin I, jasmolin II, pyrethrin I, and pyrethrin II)

3

Quinalphos 0.05

Quintozene [sum of quintozene, pentachloroaniline and methyl pentachlorophenyl sulfide (MPCPS)]

1

S-421 0.02

Tecnazene 0.05

Tetradifon 0.3

Vinclozolin 0.4

DescriptionThe sample extraction and clean-up are based on the QuEChERS method.3-9 The final extract is split for the analysis of GC- and LC-amenable pesticides using gas chromatography-tandem mass spectrometry (GC-MS/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. The reporting limits are based on the limits specified in the USP and EP pesticide residue monographs:

***Bromide and dithiocarbamates (EBDCs) are determined using different methods.


VITAMIN A, RETINOL

VITAMIN C

PurposeApplicable to the determination of retinol in foods, food products, infant formulas, premixes, pharmaceuticals, pet foods, feeds and other matrices. The method measures all-trans and 13-cis retinol isomers. It does not include contributions of vitamin A activity from pro-vitamin A carotenoids.

Method facts Sample size: 30 g in food products, 10 g in

supplements and premixes, 50 g in feeds Limit of quantitation: 1 IU/g but can vary with

sample size Precision: Most matrices - 100 IU/100 g Method reference: Official Methods of Analysis of

AOAC International, Current Ed., Methods 992.04, 992.06 and 2001.13, AOAC INTERNATI0NAL, Gaithersburg, MD, USA (modified)

DescriptionThe sample is saponified to break down fat and release the vitamins. The digest is extracted with an organic solvent. Vitamin A is quantitated directly as all-trans retinol and 13-cis retinol by ultra or high performance liquid chromatography (UHPLC or HPLC).

PurposeApplicable for the determination of vitamin C in most foods, dietary supplements and feeds. It measures both ascorbic acid and erythorbic acid if present.

Method facts Sample size: 8 g Limit of quantitation: Most matrices - 1 mg/100 g Precision: On a cereal sample matrix, the RSD is 7.33% Method reference: AOAC 967.22

DescriptionThe vitamin C in the sample is extracted,oxidized and reacted with o-phenylenediamine to produce a fluorophor. The vitamin C content is determined by comparing the sample extract fluorescence to the fluorescence of known standards.

VITAMIN C BY HPLC

PurposeApplicable for the analysis of ascorbic acid in tablets, capsules, pre-mixes, juices and raw materials.

Method facts Sample size: 1 g Limit of quantitation: Most matrices - 2 mg/g Precision: On a multivitamin sample, the RSD

is 2.44% Method reference: AOAC 967.22

Asami, D.K. (2003). Journal of Agricultural and Food Chemistry. Vol. 51. p. 1237-1241

DescriptionThe sample is extracted with an m-phosphoric acid and acetic acid solution. The extract is then analyzed on an HPLC system equipped with a UV detector. A calibration curve is used for quantification.


VITAMIN D BY LC-MS/MS

VITAMIN E (ALPHA TOCOPHEROL)

VITAMIN K1

PurposeApplicable to pet food, cereals and other matrices.

Method facts Sample size: 10 g per assay Limit of quantitation: D3 in pet food is 0.05 IU/g; D2

in pet food about 0.2 IU/g Precision: The RSD is 5% Method reference: Covance internal method

DescriptionThe sample is extracted into hexane using BHT as a preservative. A portion of the hexane is washed with ethanol/water and centrifuged. The solvent is taken to dryness and reconstituted in ACN/water and filtered through a 0.45 m PTFE filter. Isotope internal standard is added for LC-MS/MS analysis. The response is compared to that of known standards.

PurposeApplicable for the determination of vitamin E in foods, dietary supplements and premixes.

Method facts Sample size: 10 g Limit of quantitation: 0.5 IU/100 g Precision: On a corn oil matrix, the RSD is about 5% Method reference: Covance internal method

DescriptionThe product is saponified with alcoholic KOH.The tocopherols are extracted with organic solvent, separated on an HPLC silica column, and measured using fluorescence detection. Milligrams of alpha tocopherol are converted to international units (IU) by the following calculations:

1 mg d-alpha tocopherol = 1.49 IU of vitamin E(natural vitamin E)

1 mg dl-alpha tocopherol = 1.10 IU of vitamin E(synthetic vitamin E)

PurposeApplicable to the determination cis, trans, or totalvitamin K1 in various dietary supplements, infant formula, foods, drink mixes, agricultural products and premixes.

Method facts Sample size: Anywhere from 5 g to 20 tablets Limit of quantitation: 5 mcg/100 g Precision: RSD is 5.5% on infant formula Method reference: AOAC 982.27, 999.15, or USP

DescriptionThe extraction is conducted in accordance with the type of matrix being analyzed. The applicable technique from AOAC method 999.15, 982.27, or the USP is applied.The filtered extract is injected on a reverse phase HPLCsystem with post column reduction and fluorescencedetection.

Covance is an independently held company with headquarters in Princeton, New Jersey, USA.

Covance is the marketing name for Covance Inc. and its subsidiaries around the world.

The Americas + 1.888.COVANCE + 1.609.452.4440

+ 1.888.268.2623

Europe/Africa + 00.800.2682.2682

+44.1423.500888

Asia Pacific + 800.6568.3000

+65.6.5686588

[email protected]

Copyright 2014 Covance Inc.

CATNCFS002-1014

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FOOD & FOOD SUPPLEMENTS - covance.com · Method reference: Official Methods of Analysis of AOAC INTERNATIONAL, Current Ed., Method 2005.07, AOAC INTERNATIONAL, Gaithersburg, ... These