GB 5009.24-2016 PDF EnglishUS$115.00 · In stock · Download in 9 seconds
GB 5009.24-2016: National food safety standard - Determination of M Aflatoxins 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 5009.24: Historical versions
Similar standardsGB 5009.24-2016: National food safety standard - Determination of M Aflatoxins 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/GB5009.24-2016GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA National Food Safety Standard - Determination of M Aflatoxins 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... 4 1 Scope... 5 Method I -- Isotope Dilution Liquid Chromatography - Tandem Mass Spectrometry... 5 2 Principle... 5 3 Reagents and Materials... 5 4 Instruments and Apparatuses... 7 5 Analytical Procedure... 8 6 Expression of Analytical Result... 13 7 Precision... 13 8 Others... 13 Method II -- High-performance Liquid Chromatography... 14 9 Principle... 14 10 Reagents and Materials... 14 11 Instruments and Apparatuses... 15 12 Analytical Procedure... 16 13 Expression of Analytical Result... 17 14 Precision... 18 15 Others... 18 Method III -- Enzyme-linked Immunosorbent Assay... 18 16 Principle... 18 17 Reagents and Materials... 19 18 Instruments and Apparatuses... 19 19 Analytical Procedure... 19 20 Expression of Analytical Result... 20 21 Precision... 21 22 Others... 21 Appendix A Standard Concentration Calibration Method for AFT M1 and AFT M2 ... 22 Appendix B Column Capacity Verification Method for Immunoaffinity Column ... 23 Appendix C Liquid Chromatography - Mass Spectrogram and Sub-ion Scan 24 Appendix D Liquid Chromatogram... 27 Appendix E Mass Determination Method of Enzyme-linked Immunosorbent Assay Kit... 28ForewordThis Standard serves as a replacement of GB 5413.37-2010 National Food Safety Standard - Determination of Aflatoxin M1 in Milk and Milk Products; GB 5009.24-2010 National Food Safety Standard - Determination of Aflatoxin M1 and B1 in Foods; GB/T 23212-2008 Determination of Aflatoxin B1, B2, G1, G2, M1, M2 Content in Milk and Milk Powder - HPLC-fluorescence Detection Method; SN/T 1664-2005 Determination of Aflatoxin M1, B1, B2, G1, G2 Content in Milk and Milk Powder. In comparison with GB 5413.37-2010, the main changes are as follows. ---The name of the standard is modified into “National Food Safety Standard - Determination of M Aflatoxins in Foods”; ---The scope of application of the methods is extended; ---The detection of aflatoxins M2 is added; ---Enzyme-linked immunosorbent assay is modified; the name of the third method is modified into enzyme-linked immunosorbent screening assay; ---Liquid chromatography-mass spectrometry is modified; ---The pretreatment method of liquid chromatography is modified; ---Immunochromatography purification fluorescence spectrophotometry is deleted. National Food Safety Standard - Determination of M Aflatoxins in Foods1 ScopeThis Standard specifies the methods for the determination of aflatoxins M1 and aflatoxins M2 (hereinafter referred to as AFT M1 and AFT M2) in foods. Method I is isotope dilution liquid chromatography - tandem mass spectrometry, which is applicable to the determination of AFT M1 and AFT M2 in milk, milk products and milk- containing special dietary food. Method II is high-performance liquid chromatography; its scope of application is the same as Method I. Method III is enzyme-linked immunosorbent screening assay, which is applicable to the screening determination of AFT M1 in milk, milk products and milk-containing special dietary food. Method I -- Isotope Dilution Liquid Chromatography - Tandem Mass Spectrometry2 PrincipleUse methanol - water solution to extract aflatoxins M1 and aflatoxins M2 in sample. Use water or phosphate buffer solution to dilute the supernatant, then, purify and enrich it through immunoaffinity column. After concentration, constant-volume and filtering of the purified liquid, separate it through liquid chromatography. Detect it through tandem mass spectrometry, then, quantify it through isotope internal standard method.3 Reagents and MaterialsUnless it is otherwise stipulated, all reagents adopted in this Method shall be analytical pure; water shall be Grade-1 water stipulated in GB/T 6682. 3.1 Reagents 3.1.1 Acetonitrile (CH3CN). chromatographic purity. 3.1.2 Methanol (CH3OH). chromatographic purity. 3.1.8 Hydrochloric acid (HCl). 3.1.9 Petroleum ether (CnH2n+2). boiling range. 30 °C ~ 60 °C. 3.2 Reagent Preparation 3.2.1 Ammonium acetate solution (5 mmol/L). weigh-take 0.39 g of ammonium acetate; dilute it in 1,000 mL of water, then, mix it up. 3.2.2 Acetonitrile - water solution (25 + 75). measure-take 250 mL of acetonitrile; add it to 750 mL of water, then, mix it up. 3.3 Standards 3.3.1 AFT M1 standard (C17H12O7, CAS. 6795-23-9). purity ≥ 98%, or nationally certified standard substance with the standard substance certificate. 3.3.2 AFT M2 standard (C17H14O7, CAS. 6885-57-0). purity ≥ 98%, or nationally certified standard substance with the standard substance certificate. 3.3.3 13C17-AFT M1 isotope solution (C17H14O7). 0.5 μg/mL. 3.4 Standard Solution Preparation 3.4.1 Standard stock solution (10 μg/mL). respectively weigh-take 1 mg (accurate to 0.01 mg) of AFT M1 and AFT M2; respectively use acetonitrile to dissolve it, then, dilute to the constant volume of 100 mL. 3.4.2 Mixed standard stock solution (1.0 μg/mL). respectively and accurately absorb 1.00 mL of 10 μg/mL AFT M1 and AFT M2 standard stock solution, then, place them in the same 10 mL volumetric flask. 3.4.5 5 ng/mL isotope internal standard working solution 2(13C17-AFT M1). take 100 μL of AFT M1 isotope internal standard (0.5 μg/mL), then, use acetonitrile to dilute to 10 mL. Preserve it at -20 °C; it shall be used for the determination of solid sample. It shall remain valid for 3 months.4 Instruments and Apparatuses4.1 Balance. division value. 0.01 g, 0.001 g and 0.00001 g. 4.2 Water bath kettle. temperature controlled at 50 °C ± 2 °C. 4.3 Vortex mixer. 4.8 Nitrogen blowing instrument. 4.9 Liquid chromatography - tandem mass spectrometer. equipped with electrospray ion source. 4.10 Round sieve. 1 mm ~ 2 mm aperture. 4.11 Glass fiber filter paper. rapid, high-load, particle retention in liquid. 1.6 μm. 4.12 Disposable microporous filter head. equipped with 0.22 μm microporous membrane (before use, selected filter membrane shall adopt standard solution to confirm that there is no absorption phenomenon).5 Analytical ProcedureThe application of immunoaffinity columns provided by different manufacturers might be slightly different in sample operation, such as sample loading, leaching and elution. Thus, operation shall comply with the requirements in the operating instructions provided by the manufacturers. 5.1 Sample Extraction 5.1.1 Liquid milk and yogurt Weigh-take 4 g (accurate to 0.001 g) of evenly mixed sample, then, place it in a 50 mL centrifuge tube. Add 100 μL of 13C17-AFT M1 internal standard solution (5 ng/mL); start oscillation blending, then, evenly place it for 30 min. Add 10 mL of methanol, then, start vortex for 3 min. Place it at 4 °C; start centrifugation at 6,000 r/min for 10 min, or, filter it through glass fiber filter paper. Transfer an appropriate amount of the supernatant or filtrate to a beaker, then, add 40 mL of water or PBS to dilute it; reserve it for later use. 5.1.3 Cream Weigh-take 1 g (accurate to 0.001 g) of sample, then, place it in a 50 mL centrifuge tube. Add 100 μL of 13C17-AFT M1 internal standard solution (5 ng/mL); start oscillation blending, then, evenly place it for 30 min. Add 8 mL of petroleum ether; wait till cream dissolves, then, add 9 mL of water and 11 mL of methanol. Start oscillation for 30 min. Transfer all the liquid to separating funnel. Add 0.3 g of sodium chloride, then, thoroughly shake and dissolve it. Evenly place it and wait for layering, then, transfer the lower layer to a round-bottomed flask. Start rotary evaporation, till it is below 10 mL, then, use PBS to dilute it to 30 mL. 5.2 Purification 5.2.1 Preparation of immunoaffinity column Restore immunoaffinity column, which is preserved at low temperature, to room temperature. 5.2.2 Purification After abandoning the liquid in the immunoaffinity column, transfer the above-mentioned sample solution to a 50 mL syringe; 5.3 Liquid Chromatography Reference Conditions Liquid chromatography reference conditions are listed below. 5.6 Draw a Standard Curve Under the liquid chromatography - tandem mass spectrometry analytical conditions in 5.3 and 5.4, inject the standard series of solution for determination from low concentration to high concentration. Use the peak area ratio - concentration of AFT M1 and AFT M2 chromatographic peak and internal standard chromatographic peak 13C17- AFT M1 to draw the curve. Obtain standard curve regression equation; its linear correlation coefficient shall be more than 0.99. 5.8 Blank Test Do not weigh or take sample; comply with the steps in 5.1 and 5.2 to conduct the blank test. Make sure that there is no substance that would interfere the component to be determined.6 Expression of Analytical ResultThe residue of AFT M1 or AFT M2 in the sample shall be calculated in accordance with Formula (1).7 PrecisionThe absolute difference of two independent determination results obtained under repeatability conditions shall not exceed 20% of the arithmetic mean value.8 OthersWhen weighing-taking 4 g of liquid milk and yogurt, this Method’s detection limit of AFT M1 is 0.005 μg/kg; the detection limit of AFT M2 is 0.005 μg/kg; the quantitation limit of AFT M1 is 0.015 μg/kg; the quantitation limit of AFT M2 is 0.015 μg/kg.9 PrincipleUse methanol - water solution to extract aflatoxins M1 and aflatoxins M2 in sample. After diluting the supernatant, purify and enrich through the immunoaffinity column. After concentration, ......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 5009.24-2016 be delivered?Answer: The full copy PDF of English version of GB 5009.24-2016 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 5009.24-2016_English with my colleagues?Answer: Yes. The purchased PDF of GB 5009.24-2016_English will be deemed to be sold to your employer/organization who actually paid for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. Our tax invoice, downloaded/delivered in 9 seconds, includes all tax/VAT and complies with 100+ countries' tax regulations (tax exempted in 100+ countries) -- See Avoidance of Double Taxation Agreements (DTAs): List of DTAs signed between Singapore and 100+ countriesQuestion 4: Do you accept my currency other than USD?Answer: Yes. www.ChineseStandard.us -- GB 5009.24-2016 -- Click this link and select your country/currency to pay, the exact amount in your currency will be printed on the invoice. Full PDF will also be downloaded/emailed in 9 seconds.Question 5: Should I purchase the latest version GB 5009.24-2016?Answer: Yes. Unless special scenarios such as technical constraints or academic study, you should always prioritize to purchase the latest version GB 5009.24-2016 even if the enforcement date is in future. Complying with the latest version means that, by default, it also complies with all the earlier versions, technically.How to buy and download a true PDF of English version of GB 5009.24-2016?A step-by-step guide to download PDF of GB 5009.24-2016_EnglishStep 1: Visit website https://www.ChineseStandard.net (Pay in USD), or https://www.ChineseStandard.us (Pay in any currencies such as Euro, KRW, JPY, AUD).Step 2: Search keyword "GB 5009.24-2016". Step 3: Click "Add to Cart". If multiple PDFs are required, repeat steps 2 and 3 to add up to 12 PDFs to cart. Step 4: Select payment option (Via payment agents Stripe or PayPal). Step 5: Customize Tax Invoice -- Fill up your email etc. Step 6: Click "Checkout". Step 7: Make payment by credit card, PayPal, Google Pay etc. After the payment is completed and in 9 seconds, you will receive 2 emails attached with the purchased PDFs and PDF-invoice, respectively. Step 8: Optional -- Go to download PDF. Step 9: Optional -- Click Open/Download PDF to download PDFs and invoice. See screenshots for above steps: Steps 1~3 Steps 4~6 Step 7 Step 8 Step 9 |
