AOAC Official Method 2000.01 Determination of 3-Chloro-1,2-Propanediol in Foods and Food Ingredients Gas Chromatography/Mass Spectrometric Detection First Action 2000

[The method is applicable to the determination of 3-chloro-1,2-propanediol (3-monochloropropane-1,2-diol; 3-MCPD) in hydrolyzed vegetable protein (HVP), soups and stocks, stock cubes, soy sauce, malt extract, salami, fish, cheese, flour, starch, cereals, and bread.]
Caution:    This work should be performed under a fume hood. Wear laboratory coat, gloves, and eye/face protection. Use double-containment systems for handling concentrated solutions of 3-MCPD-d₅. Take care to avoid ignition of flammable reagents by sparks or static discharge.
See Table 2000.01 for the results of the interlaboratory study supporting acceptance of the method.

A. Principle

Internal standard 3-chloro-1,2-propanediol-d₅ (3-MCPD-d₅) is added to the test portion, followed by salt solution, and the mixture is blended to a homogeneous consistency. After sonication, the contents of an Extrelut^TM refill pack are added and mixed thoroughly. The mixture is transferred to a glass chromatographic tube, and the nonpolar components are eluted with a mixture of hexane and diethyl ether. The 3-MCPD is eluted with diethyl ether, and the extract is concentrated to a small volume. A portion of the concentrated extract is derivatized and analyzed by gas chromatography with mass spectrometric detection (GC/MS). The concentration of 3-MCPD is expressed in mg/kg.

AOAC Official Method 2002.05 Determination of Cholecalciferol (Vitamin D3) in Selected Foods Liquid Chromatography First Action 2002

A. Principle

After the addition of an internal standard (vitamin D₂) and basic hydrolysis, vitamin D₃ is extracted with n-heptane. The fraction that contains vitamin D₂/D₃ is separated by preparative normal-phase liquid chromatography (LC). After evaporation and dilution in acetonitrile–methanol, vitamin D₃ is determined by reversed-phase LC with UV detection at 265 nm. A separate test portion is analyzed in parallel to confirm the absence of endogenous vitamin D₂.

AOAC Official Method 2002.04 Amylase-Treated Neutral Detergent Fiber in Feeds Using Refluxing in Beakers or Crucibles First Action 2002

A. Principle

Fiber in feeds is a nutritionally defined entity that represents the indigestible and slowly digesting fraction of feeds. Neutral detergent (ND) solution and heat-stable -amylase are used to dissolve easily digested proteins, lipids, sugars, starches, and pectins in feeds, leaving a fibrous residue that is primarily cell wall components in plant materials (cellulose, hemicellulose, and lignin) and indigestible nitrogenous matter in animal products. aNDF is a gravimetric method that best estimates the total insoluble fiber, and is inversely related to digestibility and intake potential of a feed. ND soluble matter is almost completely digested by most animals; however, the digestibility of aNDF is variable among feeds and is related to lignin and other constituents in fiber.
Sodium lauryl sulfate, an anionic detergent, and sodium sulfite are used to solubilize nitrogenous matter; EDTA is used to chelate calcium and enhance removal of pectins at boiling temperatures; triethylene glycol helps remove some nonfibrous matter from concentrated feeds; disodium phosphate and sodium borate are used as buffers to maintain a neutral pH.
Hot ND solution has limited ability to solubilize starch; therefore, a heat-stable amylase is used to hydrolyze starch to saccharides that can be easily removed from fiber by filtration. Heat-stable amylases are used in hot solutions to inactivate potential contaminating enzymes in crude amylase extracts that might degrade fibrous constituents. To ensure that the amylase activity is sufficient to remove most starch and to reduce filtering difficulties, the amount of any specific amylase source needed to measure aNDF is determined under the conditions of the aNDF method.
Although boiling ND solution dissolves most proteins, lipids, and nonfibrous carbohydrates, these constituents are nonviscous only in water that is near boiling temperature. Therefore, boiling water is needed for washing fibrous residues and removing nonfibrous matter. Because fiber is particulate matter, mass-action equilibration during soaking is needed to migrate ND and contaminating soluble matter from the interior of fiber particles. Fibrous residues must be soaked, instead of being simply rinsed, in near boiling water to remove nonfibrous matter.
Boiling ND solution solubilizes lipids, and acetone soaking of the residue completes the extraction of lipids and pigments in most materials. However, excessive amounts of lipids in materials can complex with the detergent and reduce extraction efficiency. Because extraction of lipids by ND and acetone may be incomplete when feeds contain >10% lipid, these materials are pre-extracted to ensure complete removal of lipid contamination from fiber. Pre-extract materials with 5–10% lipid to minimize filtration difficulties and avoid variable aNDF results.

AOAC Official Method 2002.03 Pesticide Residues in Nonfatty Foods Supercritical Fluid Extraction and Gas Chromatography/Mass Spectrometry First Action 2002

A. Principle

A chopped frozen fruit or vegetable test portion is mixed with a drying agent, and the pesticide residue is extracted by using supercritical CO₂, collected on a solid-phase sorbent or in a liquid trap, and determined by GC/MS.

AOAC Official Method 2002.02 Resistant Starch in Starch and Plant Materials Enzymatic Digestion First Action 2002

A. Principle

Nonresistant starch is solubilized and hydrolyzed to glucose by the combined action of pancreatic -amylase and amyloglucosidase (AMG) for 16 h at 37°C. The reaction is terminated by addition of ethanol or industrial methylated spirits (IMS) and RS is recovered as a pellet by centrifugation. RS in the pellet is dissolved in 2M KOH by vigorously stirring in an ice–water bath. This solution is neutralized with acetate buffer and the starch is quantitatively hydrolyzed to glucose with AMG. Glucose is measured with glucose oxidase–peroxidase reagent (GOPOD), which is a measure of RS content. Nonresistant starch (solubilized starch) is determined by pooling the original supernatant and the washings and measuring the glucose content with GOPOD.

AOAC Official Method 2002.01 Measurement of alpha.gif (53 bytes)-Amylase Activity in White Wheat Flour, Milled Malt, and Microbial Enzyme Preparations Ceralpha Assay First Action 2002

A. Principle

Test portions are extracted with salt solution or buffer at room temperature or 40°C. Extracts are clarified by centrifugation or filtration and diluted. Aliquots of diluted extract are incubated with the substrate mixture under defined conditions of pH, temperature, and time. The substrate is nonreducing end-blocked p-nitrophenyl maltoheptaoside (BPNPG7) in the presence of excess quantities of thermostable -glucosidase. The blocking group in BPNPG7 prevents hydrolysis of this substrate by exo-acting enzymes such as amyloglucosidase, -glucosidase, or -amylase. When the substrate is cleaved by endo-acting -amylase, the nitrophenyl oligosaccharide is immediately and completely hydrolyzed to p-nitrophenol and free glucose. The reaction is terminated, and the phenolate color developed by the addition of an alkaline solution is measured at 400 nm. Results are expressed in Ceralpha units; 1 unit is defined as the amount of enzyme required to release (in the presence of excess -glucosidase) 1 µmol p-nitrophenol per minute at 40°C

44.1.12 - Sugars and Sugar Products / Sugars and Syrups

AOAC Official Method 896.02
Sucrose in Sugars and Syrups

Double Dilution Method
First Action 1896
Final Action 1970

Reference:
Analyst 21, 182(1896).