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GB/T 22224-2008: Determination of dietary fiber in foods -- Enzymatic gravimetric method and enzymatic gravimetric method -- Liquid chromatography Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid
Similar standardsGB/T 22224-2008: Determination of dietary fiber in foods -- Enzymatic gravimetric method and enzymatic gravimetric method -- Liquid chromatography---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GBT22224-2008 NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 67.050 X 04 Determination of dietary fiber in foods - Enzymatic gravimetric method and enzymatic gravimetric method-liquid chromatography Issued on. MAY 16, 2008 Implemented on. OCTOBER 01, 2008 Issued by. General Administration of Quality Supervision, Inspection and Quarantine; Standardization Administration of the People's Republic of China. Table of ContentsForeword... 4 1 Method One - Enzymatic-gravimetric method... 5 1.1 Scope... 5 1.2 Normative references... 5 1.3 Terms and definitions... 5 1.4 Method summary... 6 1.5 Reagents and solutions... 6 1.6 Apparatus and equipment... 8 1.7 Sample preparation... 9 1.8 Analysis steps... 9 1.9 Result calculation... 11 1.10 Tolerance... 12 2 Method Two - Enzymatic-gravimetric method and liquid chromatography method... 12 2.1 Scope... 12 2.2 Normative references... 13 2.3 Terms and definitions... 13 2.4 Method summary... 13 2.5 Reagents and solutions... 14 2.6 Apparatus and equipment... 14 2.7 Sample preparation... 15 2.8 Analysis steps... 15 2.9 Result calculation... 18 2.10 Tolerance... 201 Method One - Enzymatic-gravimetric method1.1 Scope Method One of this Standard specifies the conditions and detailed analysis steps for the determination of total, soluble and insoluble dietary fiber in foods by enzymatic-gravimetric method. Method One of this Standard is applicable to the determination of total, soluble and insoluble dietary fiber in cereals, vegetables and fruits as well as their products. But it is not applicable to the determination of dietary fiber in foods containing low-molecular-mass resistant maltodextrin, oligofructose, galacto- oligosaccharides, polyglucose and resistant starch. 1.2 Normative references The provisions in following documents become the provisions of this Standard through reference in this Standard. For dated references, the subsequent amendments (excluding corrigendum) or revisions do not apply to this Standard, however, parties who reach an agreement based on this Standard are encouraged to study if the latest versions of these documents are applicable. 1.3 Terms and definitions For the purposes of Method One of this Standard, the following terms and definitions apply. 1.3.1 dietary fiber the sum of carbohydrates and their analogues that have anti-digestive properties, which cannot be absorbed by the human small intestine but can be partially or completely fermented in colon 1.3.2 total dietary fiber; TDF including insoluble dietary fiber (IDF) and high-molecular-mass soluble dietary fiber (SDF) that can be precipitated in ethanol 1.4 Method summary Use thermostable α-amylase, protease and amyloglucosidase to perform enzymatic digestion of the dried sample. Use ethanol to precipitate enzymatic hydrolysate. Filter. 1.5 Reagents and solutions Unless otherwise indicated, in analysis, it shall only use the confirmed analytically-pure reagent AND distilled water or deionized water or equivalent- pure water. 1.5.1 95% ethanol. 1.5.2 Thermostable α-amylase solution. CAS 9000-85-5, IUS 3.2.1.1; cannot contain glycerol as stabilizer. Store at 0°C ~ 5°C in refrigerator. 1.5.2.1 Enzyme activity representation 1.Starch is substrate. It is represented by Nelson / Somogyi reducing sugar -- 10,000 units/mL + 1,000 units/mL (1 enzyme activity unit is defined as. at 40°C, when pH6.5, the amount of enzyme required to release 1 μmol of reducing sugar per minute). 1.5.5 Pickled diatomite. CAS 68855-54-9.Take 200 g of diatomite in 600 mL of hydrochloric acid (HCl. H2O = 1.4, volume ratio). Soak overnight. Filter. Use distilled water to wash until the filtrate is neutral. Place in 525°C ± 5°C muffle furnace to burn the ash. Leave it for use. 1.5.6 2-(N-morpholino)-sulfonate ethane (MES). CAS 4432-31-9, purity >99.5%. 1.5.7 Trihydroxymethyl aminomethane (TRIS). CAS 77-86-1, purity >99%. 1.5.8 MES-TRIS buffer (0.05 mol/L). Weigh 19.52 g of MES and 12.2 g of TRIS. Use 1.7 L of distilled water to dissolve. Use 6 mol/L sodium hydroxide to adjust pH to 8.2 ± 0.1.Add water to dilute to 2 L (attention. pH is 8.2 at 24°C; pH is 8.3 at 20°C; pH is 8.1 at 28°C. Be sure to adjust pH according to the temperature. The deviation between 20°C and 28°C is corrected by interpolation method). 1.5.9 Hydrochloric acid solution (0.561 mol/L). Take 93.5 mL of 6 mol/L hydrochloric acid. Add 700 mL of water. Mix well. Use water to set volume to 1 L. 1.5.10 Petroleum ether. Boiling range is 30°C ~ 60°C. 1.5.11 Acetone. 1.5.12 Sodium hydroxide. 1.6 Apparatus and equipment 1.6.1 High-type beaker without diversion inlet. 400 mL or 600 mL. 1.6.2 Crucible. with rough sintered glass plate; the aperture is 40 µm ~ 60 µm. Ash the cleaned crucible in muffle furnace at 525°C for 6 h. Take it out when the furnace temperature reduces below 130°C. Soak in potassium dichromate lotion for 2 h. Respectively use water and distilled water to rinse clean. In the end, use 15 mL of acetone to rinse then air-dry it. 1.6.3 Vacuum solvent filter. vacuum pump or aspirator with adjustment device. 1L suction-filtration bottle of which there is a suction-filtration mouth on the side wall; there is a rubber stopper matching with the suction-filtration bottle. It is used for the suction-filtration of enzymatic hydrolysate. 1.6.4 Constant-temperature oscillating water bath. 95°C ~ 100°C. 1.6.5 Analytical balance. resolution is 0.1 mg. 1.6.6 Balance (platform scale). measuring range is 4,000 g; resolution is 0.1 g. 1.6.7 Muffle furnace. 525°C ± 5°C. 1.6.8 Oven. 105°C, 130°C ± 3°C. 1.6.9 Vacuum drying oven. 1.6.10 Dryer. silica or equivalent desiccant. 1.6.11 pH meter. with temperature compensation function; accuracy is ±0.1. 1.6.12 Micro-Kjeldahl. 1.6.13 Pipette. 100 µL, 5 mL; disposable pipette tip. 1.7 Sample preparation 1.7.1 Food with fat content less than 10% Take well-mixed sample at 70°C vacuum to dry overnight. Place in the dryer to cool. After the dry sample is smashed, sieve through 0.3mm ~ 0.5mm sieve. If the sample cannot be heated, then lyophilize it first. Smash and sieve. Store the smashed and sieved dry sample in the dryer for use. 1.7.3 Food with high sugar content Take an appropriate amount of sample. Add 10 mL of 85% ethanol per gram of sample. Process the sample 2 ~ 3 times to de-sugar. Dry overnight at 40°C. Store the smashed-sieved dry sample in the dryer for use. 1.8 Analysis steps 1.8.1 Determination of moisture content Determine the moisture content in sample according to GB/T 5009.3-2003.Use it for result calculation. 1.8.2 Enzymatic hydrolysis 1.8.2.1 Accurately weigh double samples (ms1 and ms2), 1 g for each. The mass difference between two samples is ≤0.005 g, to the nearest of 0.1 mg. Place in 400mL or 600mL high-type beaker (1.6.1). Prepare double blank samples at the same time. Add 40 mL of pH8.2 MES-TRIS buffer into each beaker. 1.8.3 Determination 1.8.3.1 Determination of total dietary fiber 1.8.3.1.1 Precipitation. In each sample, add 225 mL (the volume after preheating) of 95% ethanol that has been preheated to 60°C. The volume ratio of ethanol to sample solution is 4.1.Take the beaker out. Cover aluminum foil. Precipitate at room temperature for 1 h. It is recommended to modify as. Weigh the mass of enzymatic hydrolysate. Use balance (1.6.6) to add 4 times mass 95% ethanol that has been preheated to 60°C. Precipitate overnight in 4°C refrigerator. 1.8.3.2 Determination of insoluble dietary fiber 1.8.3.2.1 Weigh the sample mass according to 1.8.2.1.Perform enzymatic hydrolysis according to 1.8.2.2 ~ 1.8.2.6. 1.8.3.2.2 Filtration and washing. Transfer all sample enzymatic hydrolysate to crucible to filter. 1.8.3.2.3 Determine protein and ash according to 1.8.3.1.4. 1.8.3.3 Determination of soluble dietary fiber2 Method Two - Enzymatic-gravimetric method andliquid chromatography method 2.1 Scope Method Two of this Standard specifies the conditions and detailed analysis steps for determination of total dietary fiber in foods containing low-molecular- mass resistant maltodextrin by enzymatic-gravimetric method and liquid chromatography method. 2.2 Normative references The provisions in following documents become the provisions of this Standard through reference in this Standard. For dated references, the subsequent amendments (excluding corrigendum) or revisions do not apply to this Standard, however, parties who reach an agreement based on this Standard are encouraged to study if the latest versions of these documents are applicable. 2.3 Terms and definitions For the purposes of Method Two of this Standard, the following terms and definitions apply. 2.4 Method summary Take two samples. Under the sequential actions of thermostable α-amylase, protease and amyloglucosidase, perform enzymatic hydrolysis to make starch, protein in sample into dissolved small molecules. 2.5 Reagents and solutions Unless otherwise indicated, in analysis, it shall only use the confirmed analytically-pure reagent AND distilled water or deionized water or equivalent- pure water. 2.5.1 ~ 2.5.12 Same with 1.5.1 ~ 1.5.12 of Method One. 2.6 Apparatus and equipment Laboratory routine apparatus and the followings. 2.6.1 ~ 2.6.13 Same with 1.6.1 ~ 1.6.13 of Method One. 2.6.17 Gel protection column. 6.0 mm × 40 mm, 6 μm. 2.6.18 Gel chromatographic column. 7.8 mm × 300 mm, 6 μm; two pieces. 2.7 Sample preparation 2.7.1 Same with 1.7.1 of Method One. 2.7.2 Same with 1.7.2 of Method One. But the foods that contain high sugar content may not be de-sugared. 2.8 Analysis steps 2.8.1 Determination of moisture content Same with 1.8.1 of Method One. 2.8.2 Enzymatic hydrolysis 2.8.2.4 Same with 1.8.2.4 of Method One. 2.8.2.5 Same with 1.8.2.5 of Method One. 2.8.2.6 Same with 1.8.2.6 of Method One. The amount of amyloglucosidase used is 300 μL. 2.8.3 Enzymatic gravimetric method It is used to determine the total content of insoluble dietary fiber (IDF) and high- molecular-mass soluble dietary fiber (SDF). 2.8.3.1 Precipitation. same with 1.8.3.1.1. 2.8.3.2 Filtration. same with 1.8.3.1.2. 2.8.3.3 Washing. Use 20 mL of 78% ethanol to wash high-type beaker 3 times. Under vacuum conditions, wash off the residues in crucible. 2.8.3.6 Determination of ash. Take one residue from double samples. Determine the ash in the sample according to GB/T 5009.4-2003. 2.8.3.7 Determination of protein. Take the other residue from double samples. Determine the protein content in the sample according to GB/T 5009.5-2003. 2.8.4 Liquid chromatography Use high performance liquid chromatography to determine the content of low- molecular-mass of resistant maltodextrin in sample. ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of English version of GB/T 22224-2008 be delivered?Answer: The full copy PDF of English version of GB/T 22224-2008 can be downloaded in 9 seconds, and it will also be emailed to you in 9 seconds (double mechanisms to ensure the delivery reliably), with PDF-invoice.Question 2: Can I share the purchased PDF of GB/T 22224-2008_English with my colleagues?Answer: Yes. 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