GB 5009.86-2025 English PDFUS$279.00 · In stock
Delivery: <= 3 days. True-PDF full-copy in English will be manually translated and delivered via email. GB 5009.86-2025: National food safety standard - Determination of ascorbyl palmitate in food Status: Valid GB 5009.86: Historical versions
Basic dataStandard ID: GB 5009.86-2025 (GB5009.86-2025)Description (Translated English): National food safety standard - Determination of ascorbyl palmitate in food Sector / Industry: National Standard Classification of Chinese Standard: C53 Word Count Estimation: 14,176 Date of Issue: 2025-09-02 Issuing agency(ies): National Health Commission; State Administration for Market Regulation GB 5009.86-2025: National food safety standard - Determination of ascorbyl palmitate in food---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/GB5009.86-2025National Standards of the People's Republic of China National Food Safety Standards Determination of ascorbic acid in food Published on 2025-09-02 Implemented on 2026-03-02 National Health Commission of the People's Republic of China State Administration for Market Regulation issued ForewordThis standard replaces GB 5413.18-2010 "Determination of Vitamin C in Infant Foods and Dairy Products" and GB 5009.86-2016 "Food..." Determination of ascorbic acid in the middle. Compared with GB 5009.86-2016, the main changes in this standard are as follows. ---This expands the applicability of liquid chromatography and fluorescence spectrophotometry; ---The limits of detection, limits of quantitation, and concentration ranges of standard curves for liquid chromatography and fluorescence spectrophotometry have been revised; ---The chromatographic conditions for liquid chromatography have been optimized. National Food Safety Standards Determination of ascorbic acid in food 1.Scope This standard specifies the method for determining ascorbic acid in food. The first method, liquid chromatography, is suitable for the determination of L-ascorbic acid and D-isoascorbic acid in food. The second method, fluorescence spectrophotometry, is suitable for the determination of ascorbic acid in special dietary foods, milk and dairy products, fruits and vegetables and their products. The determination of L-ascorbic acid in the above-mentioned foods without added D-isoascorbic acid was also performed. Method 3, the 2,6-dichlorophenolindophenol titration method, is suitable for the determination of reduced ascorbic acid in fruits, vegetables, and their products. First method. liquid chromatography 2.Principle After extraction, reduction, and dilution, ascorbic acid in the sample was used as the mobile phase in a mixed solution containing ion-pairing reagents and reducing agents. Reversed-phase chromatography separation, ultraviolet detector or diode array detector detection, qualitative analysis based on the retention time of the chromatographic peak, and quantitative analysis using the external standard method. 3.Reagents and Materials Unless otherwise stated, all reagents used in this method are of analytical grade, and the water is Grade I water as specified in GB/T 6682. 3.1 Reagents 3.1.1 Glacial acetic acid (C2H4O2). 3.1.2 Trichloroacetic acid (C2HCl3O2,TCA). 3.1.3 Metaphosphoric acid (HPO3). 3.1.4 40% tetrabutylammonium hydroxide aqueous solution (40% TBAH). 3.1.5 Ammonium acetate (C2H7NO2). 3.1.6 L-cysteine (C3H7NO2S). 3.1.7 Tris(2-carboxyethyl)phosphine (C9H15O6P,TCEP). 3.1.8 Acetonitrile (C2H3N). chromatographic grade. 3.2 Reagent Preparation 3.2.1 Ammonium acetate solution (500 mmol/L, pH 5.4). Weigh 19.27 g of ammonium acetate, dissolve in water and dilute to 500 mL, then use glacial acetic acid. Adjust the pH to 5.4. 3.2.2 TCA solution (150g/L). Weigh 75g of TCA, dissolve it in water and dilute it to 500mL, then mix well. 3.2.3 Metaphosphoric acid solution (200g/L). Weigh 100g of metaphosphoric acid, dissolve it in water and dilute it to 500mL, then mix well. 3.2.4 TCEP solution (250 mg/L). Weigh 125 mg of TCEP, dissolve and dilute with water to 500 mL, and mix well. 3.2.5 L-cysteine solution (40g/L). Weigh 40g of L-cysteine, add an appropriate amount of water and sonicate to dissolve, then add water to dilute to 1L and mix well. Prepare fresh before use. 3.2.6 Mobile phase. Measure 900 mL of water into a beaker, add 3.25 mL of 40% TBAH solution, and weigh 3.85 g of ammonium acetate into the water. Then add 50 mg of TCEP (or 20 mL of L-cysteine solution), followed by 10 mL of acetonitrile. Mix well and then add glacial acetic acid (approximately...). Adjust the pH to 4.5-5.0 with 3.14 mL of water; finally, dilute with water to 1 L and mix well; remove air bubbles by sonication. 3.3 Standard Products 3.3.1 L-Ascorbic Acid Standard (C6H8O6, CAS No.. 50-81-7). Purity ≥ 99%, or certified by the state and granted a standard substance certificate. Standard edition of the book. 3.3.2 D-Isoascorbic acid standard (C6H8O6, CAS No.. 89-65-6). purity ≥99%, or certified by the state and granted the title of standard substance. Standard products for certificates. 3.4 Preparation of Standard Solutions 3.4.1 L-Ascorbic Acid Standard Solution (50 μg/mL). Weigh 10 mg of L-ascorbic acid standard (accurate to 0.1 mg), and dissolve in TCEP solution. Dissolve and dilute to 10 mL with the solution, and mix well. Accurately pipette 500 μL of the above standard solution into a 10 mL amber volumetric flask, and dilute with TCEP solution. Make up to volume and mix well. Transfer the solution to a brown glass container and prepare fresh before use. 3.4.2 D-isoascorbic acid standard solution (50 μg/mL). Weigh 10 mg of D-isoascorbic acid standard (accurate to 0.1 mg), and use... Dissolve and dilute the TCEP solution to 10 mL, then mix well. Accurately pipette 500 μL of the above standard solution into a 10 mL amber volumetric flask. Make up to volume with TCEP solution and mix well. Transfer the solution to a brown glass container and prepare fresh before use. 3.4.3 L-Ascorbic Acid or D-Isoascorbic Acid Standard Series Working Solutions. Accurately pipette L-ascorbic acid or D-isoascorbic acid standard solutions respectively. Dissolve 10 μL, 100 μL,.200 μL, 400 μL, 1000 μL, and.2000 μL of a 50 μg/mL solution in 10 mL brown volumetric flasks and pump the solutions using a flow purifier. Make up to volume and mix gently. The mass concentrations of the L-ascorbic acid or D-isoascorbic acid standard series working solutions are 0.05 μg/mL, respectively. 0.50 μg/mL, 1.0 μg/mL, 2.0 μg/mL, 5.0 μg/mL and 10 μg/mL, prepare fresh before use. Note. The concentration range of the standard solution can be adjusted appropriately within the linear range based on the actual sample to be tested. 4.Instruments and Equipment 4.1 Liquid Chromatograph. Equipped with an ultraviolet detector or a diode array detector. 4.2 Balance. Sensitivity is 0.01 mg and 0.1 mg respectively. 4.3 pH meter. accuracy 0.1. 4.4 Vortex mixer. rotation speed not less than 1500 r/min. 4.5 Centrifuge. speed not less than 5000 r/min. 4.6 Ultrasonic cleaner. 5.Analysis Steps 5.1 Sample Preparation Liquid and slurry samples should be shaken thoroughly; semi-solid and powdered samples should be mixed thoroughly; gelatinous samples such as gum-based candies should be cut into small pieces with scissors or frozen. For other samples, homogenization or pulverization is required. Fruits and vegetables and their products (edible portions) should be added at a ratio of 10.1 to 150 g/L. Homogenize or grind the solution with TCA or.200 g/L metaphosphoric acid solution. Note. The process should be carried out under light-protected conditions as much as possible; the next test should be conducted immediately after all samples are prepared. 5.1.1 Sample Weighing 5.1.1.1 Solid (including semi-solid) samples Weigh 25g (accurate to 0.001g) of the prepared sample into an Erlenmeyer flask, add 225g (accurate to 0.001g) of TCEP solution, and fill the flask. Dissolve and mix thoroughly. Weigh 2g (accurate to 0.001g) of the mixed solution into a 10mL volumetric flask. 5.1.1.2 Liquid (including slurry) samples Weigh 2g (accurate to 0.001g) of the prepared sample or accurately pipette 2mL into a 10mL volumetric flask. 5.1.2 Sample Extraction and Reduction Add 4 mL of TCEP solution (or L-cysteine solution) to the 10 mL volumetric flask mentioned above, then add 2 mL of TCA solution and mix. Mix well, let stand for 5 minutes, dilute to volume with water, and mix thoroughly. Then transfer to a centrifuge tube and centrifuge at 5000 rpm for 1 minute. This sample solution is the reduced sample solution. Note. Special medical purpose formula foods such as amino acid formula powder, hydrolyzed protein formula powder, and partially hydrolyzed protein formula powder, as well as goat milk powder, should use L-cysteine as the active ingredient. It is a reducing agent. 5.1.3 Sample dilution Transfer 1 mL of the supernatant from centrifugation to a 10 mL volumetric flask, add 1 mL of ammonium acetate solution, dilute to volume with the mobile phase, and mix well. To dilute the sample solution, take an appropriate amount of the solution and pass it through a 0.22 μm aqueous microporous membrane into a brown sample vial for liquid chromatography determination. Note. If necessary, the dilution factor can be changed to ensure that the concentration of ascorbic acid in the test solution is within the linear range. 5.2 Instrument Reference Conditions 5.2.1 Chromatographic column. C18 column (150 mm in length, 4.6 mm in inner diameter, and 4 μm in particle size) or a column with equivalent performance. 5.2.2 Column temperature. room temperature. 5.2.3 Mobile phase. Prepared according to 3.2.6. 5.2.4 Flow rate. 1.0 mL/min. 5.2.5 Detection wavelength. 265nm. 5.2.6 Injection volume. 10 μL. 5.3 Construction of Standard Curve The standard series of working solutions were injected into the liquid chromatograph to obtain the corresponding peak areas. The L-ascorbic acid content in the standard series of working solutions was then used as the reference. A standard curve was plotted with the concentration of L-ascorbic acid or D-isoascorbic acid on the x-axis and the peak area on the y-axis. The chromatogram of the standard solution is shown in Figure A.1 in Appendix A. 5.4 Determination of Sample Solution The sample solution was injected into the liquid chromatograph to obtain the peak area of ascorbic acid. The ascorbic acid content in the test solution was then determined based on the standard curve. concentration. 6.Presentation of Analysis Results The content of L-ascorbic acid or D-isoascorbic acid in the sample is calculated according to formula (1). X=ρ ×M0×V1×V3×f×100 m0×m×V2×1000 (1) ......Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of GB 5009.86-2025_English be delivered?Answer: Upon your order, we will start to translate GB 5009.86-2025_English as soon as possible, and keep you informed of the progress. The lead time is typically 1 ~ 3 working days. The lengthier the document the longer the lead time.Question 2: Can I share the purchased PDF of GB 5009.86-2025_English with my colleagues?Answer: Yes. The purchased PDF of GB 5009.86-2025_English will be deemed to be sold to your employer/organization who actually pays for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. 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