GB 5009.154-2023 PDF EnglishUS$380.00 · In stock · Download in 9 seconds
GB 5009.154-2023: National food safety standard - Determination of vitamin B6 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 5009.154: Historical versions
Similar standardsGB 5009.154-2023: National food safety standard - Determination of vitamin B6 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/GB5009.154-2023GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National Food Safety Standard - Determination of Vitamin B6 in Foods Issued on. SEPTEMBER 6, 2023 Implemented on. MARCH 6, 2024 Issued by. National Health Commission of the People’s Republic of China; State Administration for Market Regulation. Table of ContentsForeword... 4 1 Scope... 5 Method I - Liquid Chromatography - Tandem Mass Spectrometry... 5 2 Principle... 5 3 Reagents and Materials... 5 4 Instruments and Equipment... 8 5 Analytical Procedures... 9 6 Expression of Analysis Results... 13 7 Precision... 14 8 Others... 14 Method II - Liquid Chromatography - Mass Spectrometry... 14 9 Principle... 14 10 Reagents and Materials... 14 11 Instruments and Equipment... 17 12 Analytical Procedures... 18 13 Expression of Analysis Results... 21 14 Precision... 22 15 Others... 22 Method III - High-performance Liquid Chromatography - Fluorescence Detection Method... 22 16 Principle... 22 17 Reagents and Materials... 22 18 Instruments and Equipment... 24 19 Analytical Procedures... 25 20 Expression of Analysis Results... 26 21 Precision... 27 22 Others... 27 Method IV - Microbiological Method... 271 ScopeThis Standard specifies the method for the determination of vitamin B6 in foods. The first method “liquid chromatography - tandem mass spectrometry” is applicable to the determination of vitamin B6 in foods. The second method “liquid chromatography - mass spectrometry” is applicable to the determination of vitamin B6 (pyridoxamine, pyridoxal and pyridoxine) in formulated milk powder, special dietary foods, ready-to-eat cereals, baked goods and beverages, in which, vitamin B6 (pyridoxamine, pyridoxal and pyridoxine) is added as a nutritional fortifier. The third method “high-performance liquid chromatography - fluorescence detection method” is applicable to the determination of vitamin B6 (pyridoxamine, pyridoxal and pyridoxine) in formulated milk powder, special dietary foods (except foods for special medical purposes), ready-to-eat cereals, baked goods and beverages, in which, vitamin B6 (pyridoxamine, pyridoxal and pyridoxine) is added as a nutritional fortifier. The fourth method “microbiological method” is applicable to the determination of vitamin B6 in foods. Method I - Liquid Chromatography - Tandem Mass Spectrometry2 PrincipleVitamin B6 (pyridoxamine, pyridoxal and pyridoxine) in foods is firstly hydrolyzed by acid, then, enzymatically hydrolyzed into pyridoxamine, pyridoxal and pyridoxine. After dilution and filtration, reversed-phase liquid chromatography separation, tandem mass spectrometry detection, and isotope internal standard method for quantitative determination, the total vitamin B6 content is calculated in terms of pyridoxine.3 Reagents and MaterialsUnless it is otherwise specified, the reagents used in this Method are all analytically pure, and the water is Grade-1 water specified in GB/T 6682. 3.1 Reagents 3.2 Reagent Preparation 3.3 Reference Materials 3.4 Preparation of Standard Solutions 3.5 Preparation of Isotope Internal Standard Solutions4 Instruments and Equipment4.1 Liquid chromatography - tandem mass spectrometer. equipped with electrospray ion source. 4.3 Constant-temperature incubator, or one with equivalent performance. 4.4 pH meter. with an accuracy of 0.01. 4.5 Vortex mixer. 4.6 Spectrophotometer. 4.7 High-speed pulverizer. 4.8 Homogenizer. 4.9 Constant-temperature water bath or autoclave.5 Analytical Procedures5.1 Specimen Preparation For meat, vegetables and fruits, etc., take the edible parts, use water to wash them, and use dry gauze to wipe off the surface moisture, use a homogenizer to homogenize them and store in sample bottles for later use. For powdery samples (milk powder and rice noodles, etc.), evenly mix them and conduct direct sampling. For liquid samples, evenly shake them and reserve them for later use. For liquid samples with granules, for example, large-grained yogurt, use a homogenizer to homogenize them and reserve them for later use. For flaky and granular samples, use a high-speed pulverizer to grind them into powder and seal for later use. 5.2 Specimen Treatment 5.3 Determination Conditions of Instruments 5.4 Drawing of Standard Curve Inject the vitamin B6 mixed standard series working solutions into the liquid chromatography - mass spectrometer from low to high concentrations. Take the concentrations of the working solutions of pyridoxine, pyridoxal and pyridoxamine as the x-coordinate, and the peak area ratio of pyridoxine, pyridoxal and pyridoxamine to their corresponding isotope internal standards as the y-coordinate. Respectively draw the standard curves of pyridoxamine, pyridoxal and pyridoxine. 5.5 Determination of Specimen Solution Inject the solution to be tested. In accordance with the three standard curves drawn above, adopt the internal standard method to calculate the concentration of pyridoxine, pyridoxal and pyridoxamine in the specimen solution to be tested.6 Expression of Analysis Results6.1 The content of each component of vitamin B6 in the specimen is calculated in accordance with Formula (1). 6.2 Total content of vitamin B6 (calculated as pyridoxine) The total content of vitamin B6 in the specimen is calculated in accordance with Formula (2).7 PrecisionThe absolute difference between the results of two independent determinations obtained under repeatability conditions shall not exceed 10% of the arithmetic mean.8 Others8.1 Solid specimens. when the sampling size is 1.00 g, the detection limit of the Method is. 0.03 mg/kg for pyridoxine, 0.03 mg/kg for pyridoxal and 0.03 mg/kg for pyridoxamine; the quantitation limit is 0.10 mg/kg for pyridoxine, 0.10 mg/kg for pyridoxal and 0.10 mg/kg for pyridoxamine. 8.2 Liquid specimens. when the sampling size is 5.00 g, the detection limit of the Method is. 0.01 mg/kg for pyridoxine, 0.01 mg/kg for pyridoxal and 0.01 mg/kg for pyridoxamine;9 PrincipleVitamin B6 (pyridoxine, pyridoxal and pyridoxamine) added to foods is subject to extraction, dilution and filtration, and separation by reversed-phase liquid chromatographic column, nitrogen and single quadrupole mass spectrometry detection. Adopt the isotope dilution internal standard method to respectively quantify pyridoxamine, pyridoxal and pyridoxine. The total vitamin B6 content is calculated in terms of pyridoxine.10 Reagents and MaterialsUnless it is otherwise specified, the reagents used in this Method are all analytically pure, and the water is Grade-1 water specified in GB/T 6682. 10.1 Reagents 10.1.1 Methanol (CH3OH). chromatographically pure. 10.1.2 Formic acid (HCOOH). chromatographically pure. 10.1.3 Ammonium formate (HCOONH4). chromatographically pure. 10.1.4 Hydrochloric acid (HCl). 10.1.5 Sodium hydroxide (NaOH). 10.2 Reagent Preparation 10.2.1 Hydrochloric acid solution (0.1 mol/L). accurately draw 9 mL of hydrochloric acid and use water to dilute to 1,000 mL. 10.2.2 Sodium hydroxide solution (0.1 mol/L). accurately weigh-take 0.4 g of sodium hydroxide, add 50 mL of water to dissolve it; after cooling, use water to dilute to 100 mL. 10.3 Reference Materials 10.4 Preparation of Standard Solutions 10.4.5 Vitamin B6 standard mixed working solution (5.00 g/mL). accurately draw 1.00 mL of vitamin B6 standard mixed stock solution (50.0 g/mL) into a 10 mL brown volumetric flask, use mobile phase A to dilute to the scale. Prepare it right before use. 10.5 Preparation of Isotope Internal Standard Solutions 10.6 Preparation of Standard Series of Working Solutions Respectively and accurately draw an appropriate amount of vitamin B6 standard mixed working solution into a 50 mL brown volumetric flask, and accurately add 100 L of vitamin B6 isotope internal standard mixed working solution (50.0 g/mL). Use mobile phase A to dilute to the scale. The mass concentration of the vitamin B6 (pyridoxamine, pyridoxal and pyridoxine) standard series working solutions is respectively 10.0 ng/mL, 20.0 ng/mL, 40.0 ng/mL, 100 ng/mL, 200 ng/mL and 250 ng/mL. Their corresponding isotope internal standard concentrations are all 100 ng/mL. Prepare them right before use.11 Instruments and Equipment11.1 Liquid chromatography - mass spectrometer. equipped with electrospray ion source. 11.2 Balance. with a division value of 0.01 g and 0.1 mg. 11.3 Constant-temperature incubator, or one with equivalent performance. 11.4 pH meter. with an accuracy of 0.01. 11.8 Homogenizer. 11.9 Constant-temperature water bath.12 Analytical Procedures12.2 Specimen Treatment 12.3 Determination Conditions of Instruments 12.3.1 Reference conditions of chromatography The reference conditions of chromatography are as follows. 12.3.2 Reference conditions of mass spectrometry The reference conditions of mass spectrometry are as follows. 12.4 Drawing of Standard Curve Inject the vitamin B6 (pyridoxine, pyridoxal and pyridoxamine) mixed standard series working solutions into the liquid chromatography - mass spectrometer from low to high concentrations. Take the concentrations of the working solutions of pyridoxine, pyridoxal and pyridoxamine as the x-coordinate, and the peak area ratio of pyridoxine, pyridoxal and pyridoxamine to their corresponding isotope internal standards as the y-coordinate. Respectively draw the standard curves of pyridoxamine, pyridoxal and pyridoxine. 12.5 Determination of Specimen Solution Inject the solution to be tested. In accordance with the three standard curves drawn above, adopt the internal standard method to calculate the concentration of pyridoxine, pyridoxal and pyridoxamine in the specimen solution to be tested. 12.6 Qualitative Determination When comparing the retention time of the chromatographic peak of the target compound in the specimen with the retention time of the corresponding standard chromatographic peak, ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. 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