GB 5009.85-2016 PDF EnglishUS$135.00 · In stock · Download in 9 seconds
GB 5009.85-2016: National food safety standard - Determination of vitamin B2 in foods Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid GB 5009.85: Historical versions
Similar standardsGB 5009.85-2016: National food safety standard - Determination of vitamin B2 in foods---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/GB5009.85-2016GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National food safety standard - Determination of vitamin B2 in foods Issued on. DECEMBER 23, 2016 Implemented on. JUNE 23, 2017 Issued by. National Health and Family Planning Commission of the PRC; China Food and Drug Administration. Table of ContentsForeword... 3 1 Scope... 4 2 Principles... 4 3 Reagents and materials... 4 4 Instrument and equipment... 6 5 Analytical procedures... 6 6 Analysis results expression... 7 7 Precision... 8 8 Others... 8 9 Principles... 8 10 Reagents and materials... 9 11 Instrument and equipment... 10 12 Analytical procedures... 11 13 Analysis results expression... 13 14 Precision... 13 15 Others... 13 Appendix A Concentration calibration method of vitamin B2 standard solution ... 14 Appendix B Liquid chromatogram of vitamin B2... 16ForewordThis standard replaces GB/T 5009.85-2003 "Determination of riboflavin in foods", GB/T 9695.28-2008 "Meat and meat products-Determination of vitamin B2 content", GB/T 7629-2008" Determination of vitamin B2 in cereals "and GB 5413.12-2010" National food safety standard-Determination of vitamin B2 in foods for infants and young children, milk and milk products". As compared with GB/T 5009.85-2003, the main changes of this standard are as follows. -- The standard name is changed to "National food safety standard- Determination of vitamin B2 in foods"; -- Add high performance liquid chromatography; -- Delete microbiological method. National food safety standard - Determination of vitamin B2 in foods1 ScopeThis standard specifies the determination of vitamin B2 in foods. The first method of this standard is high performance liquid chromatography and the second method is fluorescence spectrophotometry, which apply to the determination of vitamin B2 in various type of foods. Method I. High performance liquid chromatography2 PrinciplesThe sample is hydrolyzed in dilute hydrochloric acid at constant temperature, adjusted to pH 6.0-6.5; use papain and taka-amylase to carry out enzymolysis. After filtered through constant-volume, the filter liquor is separated through reversed phase column and detected by HPLC fluorescence detector. Use external standard method to quantify.3 Reagents and materialsUnless otherwise indicated, the reagents used in this method are analytical pure, the water is the grade-1 water specified in GB/T 6682. 3.1 Reagents 3.1.1 Hydrochloric acid (HCl). 3.1.7 Taka-amylase. active units ≥ 100 U/mg, or similar performance. 3.2 Reagent preparation 3.2.1 Hydrochloric acid solution (0.1 mol/L). PIPETTE 9 mL of hydrochloric acid; USE water to dilute it to 1000mL. 3.2.4 Sodium acetate solution (0.1 mol/L). Accurately WEIGH 13.60 g of sodium acetate trihydrate; ADD 900 mL of water to dissolve it; USE water to dilute it to 1000 mL. 3.2.5 Sodium acetate solution (0.05 mol/L). Accurately WEIGH 6.80 g of sodium acetate trihydrate; ADD 900 mL of water to dissolve it; ADJUST the pH to 4.0 ~ 5.0 with glacial acetic acid; USE water to dilute it to 1000 mL. 3.2.6 Mixed enzyme solution. Accurately WEIGH 2.345 g of papain and 1.175g of taka-amylase; ADD water to dissolve it; DILUTE it to 50 mL. Prepare it before use. 3.3 Standards Vitamin B2 (C17H20N4O6, CAS number. 83-88-5). Purity≥98%. 3.4 Standard solution preparation 3.4.1 Vitamin B2 standard stock solution (100 μg/mL). PLACE the vitamin B2 standard in a vacuum desiccator or dryer with phosphorus pentoxide; after drying for 24h, accurately WEIGH 10 mg (accurate to 0.1 mg) Vitamin B2 standard; ADD 2 mL of hydrochloric acid solution (1+1) to dissolve it in ultrasound; 3.4.2 Vitamin B2 standard intermediate solution (2.00 μg/mL). Accurately PIPETTE 2.00 mL of Vitamin B2 standard stock solution; USE water to dilute it to 100 mL. Prepare before use.4 Instrument and equipment4.1 High performance liquid chromatograph. equipped with fluorescence detector. 4.2 Balance. sensitivities are 1 mg and 0.01 mg. 4.6 Tissue-mash-machine. 4.7 Thermostatic water bath. 4.8 Dryer. 4.9 Spectrophotometer.5 Analytical procedures5.1 Sample preparation TAKE out 500 g of sample; USE tissue-mash-machine to smash it uniformly; respectively PLACE it into clean brown grinding mouth bottle; MAKE a mark after sealing; STORE it for spare-use in dark place. 5.2 Instrument reference conditions 5.3 Drawing of standard curve LEAD the standard series working solution into high performance liquid chromatograph respectively; MEASURE the corresponding peak area. With the concentration of standard working solution as abscissa AND the peak area as vertical axis, DRAW a standard curve. 5.4 Determination of sample solution LEAD the sample solution into high performance liquid chromatograph to obtain the corresponding peak area; OBTAIN the vitamin B2 concentration of the solution under test according to the standard curve.6 Analysis results expressionThe Vitamin B2 content in the sample is calculated in accordance with the equation (1).7 PrecisionThe absolute difference between two independent determinations obtained under repeatability conditions shall not exceed 10% of the arithmetic mean.8 OthersWhen the sampling weight is 10.00 g, the method detection limit is 0.02 mg/100g and the quantitative limit is 0.05 mg/100g.9 PrinciplesExposed under the light at a wavelength of 440 nm ~ 500 nm, vitamin B2 sends out yellowish green fluorescence. In dilute solution, its fluorescence intensity is proportional to the concentration of vitamin B2.Measure its fluorescence intensity at a wavelength of 525 nm.10 Reagents and materialsUnless otherwise indicated, the reagent used in this method is analytical pure, the water is the grade-1 water specified in GB/T 6682 regulations. 10.1 Reagents 10.1.1 Hydrochloric acid (HCl). 10.1.10 Hydrogen peroxide (H2O2). 30%. 10.1.11 Sodium dithionite (Na2S2O4). 10.2 Reagent preparation 10.2.1 Hydrochloric acid solution (0.1 mol/L). PIPETTE 9 mL of hydrochloric acid; USE water to dilute it to 1000 mL. 10.2.5 Mixed enzyme solution. Accurately WEIGH 2.345 g of papain and 1.175 g of taka-amylase; ADD water to dissolve it; USE water to dilute it to 50 mL. Prepare it before use. 10.2.6 Eluent. acetone-glacial acetic acid-water (5+2+9, volume ratio). 10.2.7 Potassium permanganate solution (30 g/L). Accurately WEIGH 3 g of potassium permanganate; USE water to dissolve it and dilute it to 100 mL. 10.3 Standard products Vitamin B2 (C17H20N4O6, CAS number. 83-88-5). Purity≥98%. 10.4 Standard solution preparation 10.4.1 Vitamin B2 standard stock solution (100 μg/mL). PLACE the vitamin B2 standard in a vacuum desiccator or dryer with phosphorus pentoxide; after drying for 24h, accurately WEIGH 10 mg (accurate to 0.1 mg) Vitamin B2 standard; ADD 2 mL of hydrochloric acid solution (1+1) to dissolve it in ultrasound; immediately USE water to transfer it and dilute it to 100 mL.11 Instrument and equipment11.1 Fluorescence Spectrophotometer. 11.2 Balance. sensitivities are 1 mg and 0.01 mg. 11.6 Tissue-mash-machine. 11.7 Thermostat water bath. 11.8 Dryer. ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. 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