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GB 1903.76-2025: National food safety standard - Nutrition fortifier - Hematin
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National food safety standard - Nutrition fortifier - Hematin
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GB 1903.76-2025
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Basic data | Standard ID | GB 1903.76-2025 (GB1903.76-2025) | | Description (Translated English) | National food safety standard - Nutrition fortifier - Hematin | | Sector / Industry | National Standard | | Classification of Chinese Standard | X40 | | Word Count Estimation | 8,817 | | Date of Issue | 2025-09-02 | | Issuing agency(ies) | National Health Commission; State Administration for Market Regulation | GB 1903.76-2025: National food safety standard - Nutrition fortifier - Hematin---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/GB1903.76-2025
National Standards of the People's Republic of China
National Food Safety Standards
Food fortifier heme iron
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
National Food Safety Standards
Food fortifier heme iron
1.Scope
This standard applies to the use of animal blood that has passed inspection and quarantine or blood cells obtained through centrifugation as raw materials, which undergo enzymatic hydrolysis, separation, and drying.
The resulting food fortifier is heme iron.
2.Chemical name, molecular formula, structural formula, and relative molecular mass
2.1 Chemical Name
Heme iron
2.2 Molecular Formula
C34H32FeN4O4
2.3 Structural Formula
2.4 Relative Molecular Mass
616.499 (based on 2022 international relative atomic mass)
3 Technical Requirements
3.1 Sensory Requirements
Sensory requirements shall comply with the provisions of Table 1.
Table 1 Sensory Requirements
Project Requirements Inspection Methods
Color ranges from reddish-brown to dark brown
Powder or granules
Odorless or with a slight characteristic odor of heme iron
Impurities. None visible to the naked eye
Take an appropriate amount of sample and place it in a clean, dry, transparent glass container.
Observe its color and condition under natural light, smell its odor, and observe its impurities.
3.2 Physicochemical Indicators
The physicochemical properties should meet the requirements of Table 2.
Table 2 Physicochemical Indicators
Project indicator testing methods
Iron (as Fe), w/% 1.0~2.6 GB 5009.90
Heme iron (calculated as C34H32FeN4O4), w/% 9.0~27.0 (See Appendix A, A.3)
Loss on drying, w/% ≤ 6.0 GB 5009.3 (Direct drying method)
Residue on ignition, w/% ≤ 12.0 GB 5009.4
Lead (Pb)/(mg/kg) ≤ 2.0 GB 5009.75 or GB 5009.12
Total arsenic (as As)/(mg/kg) ≤ 2.0 GB 5009.76 or GB 5009.11
3.3 Microbial limits
Microbial limits should comply with the provisions of Table 3.
Table 3 Microbial Limits
project
Sampling scheme a and limits
ncm M
Test methods
Salmonella/25g 5 0 0 - GB 4789.4
Staphylococcus aureus/(CFU/g) 5 1 100 1000 GB 4789.10 Method II
a. The sampling and processing of samples shall be carried out in accordance with GB 4789.1.
Appendix A
Test methods
A.1 General Provisions
Unless otherwise specified, the reagents and water used in the test methods of this standard refer to analytical grade reagents and those specified in GB/T 6682.
The solution used in the experiment is a grade III water. Unless otherwise specified, all solutions used in the experiment are aqueous solutions.
A.2 Identification Test
A.2.1 Reagents and Materials
A.2.1.1 Sodium hydroxide.
A.2.1.2 Pyridine.
A.2.1.3 Sodium hyposulfite.
A.2.1.4 Nitric acid. chromatographic grade.
A.2.1.5 Ammonia water.
A.2.1.6 1mol/L sodium hydroxide solution. Weigh 40.0g of sodium hydroxide, dissolve it in carbon dioxide-free water, and bring the volume to 1L.
A.2.1.7 Pyridine sodium hydroxide solution. Pipette 100 mL of pyridine, add 30 mL of 1 mol/L sodium hydroxide solution, and dilute with water to the specified concentration.
300mL.
A.2.2 Instruments and Equipment
A.2.2.1 Analytical balance. sensitivity 0.0001g.
A.2.2.2 Ultraviolet spectrophotometer.
A.2.3 Identification Method
A.2.3.1 Color Development Test
A.2.3.1.1 Weigh 0.01 g (accurate to 0.0001 g) of heme iron sample, dissolve it in 50 mL of pyridine sodium hydroxide solution, and take...
Add 15 mg of sodium hyposulfite to 5 mL of solution; the solution should turn red.
A.2.3.1.2 Weigh 0.01 g (accurate to 0.0001 g) of heme iron sample into a 100 mL beaker, add 5 mL of nitric acid, heat, and dissolve.
The liquid should be yellow. After cooling, it should be alkalized with ammonia water, and the solution should turn orange-yellow.
A.2.3.2 Ultraviolet Spectroscopy
Weigh 0.01 g (accurate to 0.0001 g) of heme iron sample, dissolve it in 50 mL of pyridine sodium hydroxide solution, and scan under ultraviolet light.
The spectrum shows maximum absorption at a wavelength of 399 nm.
A.3 Determination of Heme Iron (C34H32FeN4O4) Content
A.3.1 Method Summary
Heme iron in the sample was dissolved in pyridine sodium hydroxide, separated by liquid chromatography, and detected by ultraviolet or diode array detector.
Qualitative analysis using time, quantitative analysis using the external standard method based on peak area.
A.3.2 Reagents and Materials
Unless otherwise specified, all reagents used are of analytical grade, and the water is Grade I water as specified in GB/T 6682.
A.3.2.1 Methanol. chromatographic grade.
A.3.2.2 Formic acid. chromatographic grade.
A.3.2.3 Sodium hydroxide.
A.3.2.4 Pyridine.
A.3.2.5 1mol/L sodium hydroxide solution. Weigh 40.0g of sodium hydroxide, dissolve it in carbon dioxide-free water, and bring the volume to 1L.
A.3.2.6 Pyridine sodium hydroxide solution. Pipette 100 mL of pyridine, add 30 mL of 1 mol/L sodium hydroxide solution, and dilute with water to the specified concentration.
300mL.
A.3.2.7 Hydroxyheme (C34H33FeN4O5, CAS No.. 15489-90-4) Standard. Content ≥98.0%.
A.3.2.8 Hydroxymethene Standard Stock Solution. Accurately weigh 0.1028 g of hydroxymethene dried at 100℃ for 3 h, and oxidize with pyridine.
Dissolve and dilute to 100 mL in sodium solution (A.3.2.6). This solution is equivalent to 1 mg/mL of heme iron. Store in a brown reagent bottle at 0°C.
Store at 4℃ away from light; shelf life is 7 days.
A.3.2.9 Filter membrane. 0.45μm, aqueous phase.
A.3.3 Instruments and Equipment
A.3.3.1 High-performance liquid chromatograph. equipped with an ultraviolet detector or a diode array detector.
A.3.3.2 Analytical balance. sensitivity 0.0001g.
A.3.3.3 Ultrasonic generator. power greater than 180W.
A.3.3.4 Centrifuge. greater than 4000 r/min.
A.3.3.5 Constant temperature drying oven.
A.3.4 Reference Conditions for Liquid Chromatography
A.3.4.1 Chromatographic column. C18 column, 250 mm in length, 4.6 mm in inner diameter, 5 μm in particle size or equivalent column.
A.3.4.2 Mobile phase. Methanol. 0.1% formic acid aqueous solution = 70. 30 (volume ratio).
A.3.4.3 Flow rate. 1.0 mL/min.
A.3.4.4 Column temperature. 30℃.
A.3.4.5 Injection volume. 5.0 μL.
A.3.4.6 Detection wavelength. 399nm.
A.3.5 Analytical Steps
Accurately weigh 0.1 g of the sample (accurate to 0.0001 g), dissolve it in pyridine sodium hydroxide solution (A.3.2.6), and sonicate under light-protected conditions.
Dissolve for 10 minutes, cool to room temperature, transfer to a 50 mL volumetric flask, dilute to the mark with pyridine sodium hydroxide solution, mix well, and measure out the solution.
1 mL was placed in a 10 mL volumetric flask, diluted to the mark with pyridine sodium hydroxide solution, centrifuged at 4000 r/min for 10 min, and the supernatant was collected and filtered through a membrane.
Filter and determine by liquid chromatography. Perform a blank test, except that the procedure is the same as for the sample test.
A.3.6 Qualitative Analysis
The qualitative identification method based on the consistency of retention time, according to the retention time of the hydroxymethylheme standard, has a deviation of ±2.5%.
Within this range, determine the chromatographic peak of heme iron in the sample (see Appendix B).
A.3.7 Quantitative Analysis
Take 0.25 mL, 0.50 mL, 1.00 mL, 2.00 mL, and 3.00 mL of hydroxymethemoglobin standard stock solution (A.3.2.8) respectively.
5.00 mL was added to a 50 mL volumetric flask and diluted to volume with pyridine sodium hydroxide solution (A.3.2.6) to prepare a solution equivalent to the mass concentration of heme iron solution.
Standard solutions with concentrations of 5 μg/mL, 10 μg/mL, 20 μg/mL, 40 μg/mL, 60 μg/mL, and 100 μg/mL were used for liquid chromatography determination.
A standard curve was plotted with heme iron concentration on the x-axis and peak area on the y-axis.
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