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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.308-2025 | English | 215 |
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GB 5009.308-2025: 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/GB5009.308-2025
GB
NATIONAL STANDARD OF THE
PEOPLE'S REPUBLIC OF CHINA
National food safety standard - Determination of ascorbyl
palmitate in food
Issued on: SEPTEMBER 2, 2025
Implemented on: MARCH 2, 2026
Issued by. National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
1 Scope... 3
2 Principle... 3
3 Reagents and materials... 3
4 Instruments and equipment... 4
5 Analysis steps... 5
6 Presentation of analysis results... 6
7 Precision... 7
8 Others... 7
Appendix A Chromatogram of ascorbyl palmitate standard solution... 8
National food safety standard - Determination of ascorbyl
palmitate in food
1 Scope
This standard specifies a liquid chromatography method for the determination of
ascorbyl palmitate in food.
This standard applies to the determination of ascorbyl palmitate in food.
2 Principle
The ascorbyl palmitate in the sample is extracted with citric acid-isoascorbic acid
methanol solution and then separated by reversed-phase liquid chromatography.
Detection is performed using a diode array detector or ultraviolet detector, and
quantification is performed 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 Methanol (CH4O). chromatographically pure.
3.1.2 Acetonitrile (C2H3N). chromatographically pure.
3.1.3 Phosphoric acid (H3PO4).
3.1.4 Citric acid (C6H8O7).
3.1.5 Isoascorbic acid (C6H8O6).
3.2 Reagent preparation
3.2.1 Citric acid-isoascorbic acid methanol solution. Weigh 1.0 g of citric acid and 1.0
g of isoascorbic acid, dissolve them in methanol and dilute to 1000 mL.
3.2.2 Phosphoric acid solution (0.1%). Measure 1.0 mL of phosphoric acid, dilute with
water to 1000 mL, mix well and set aside.
3.2.3 Methanol-acetonitrile solution (50+50). Mix methanol and acetonitrile at a
volume ratio of 50.50 and set aside.
3.3 Standard substances
3.3.1 Ascorbyl palmitate standard substance (C22H38O7, CAS No.. 137-66-6). with a
purity of ≥98%, or a standard substance that has obtained the national certification and
been granted a standard substance certificate.
3.4 Preparation of standard solutions
3.4.1 Ascorbyl palmitate standard solution (1.00 mg/mL). Accurately weigh 25 mg
(accurate to 0.1 mg) of the standard substance, dissolve it in citric acid-isoascorbic acid
methanol solution and dilute to 25 mL, then mix well. Prepare fresh before use.
3.4.2 Ascorbyl palmitate standard intermediate solution (0.100 mg/mL). Accurately
pipette 1.00 mL of the 1.00 mg/mL standard solution, dilute with citric acid-isoascorbic
acid methanol solution and bring the volume to 10 mL, then mix well. Prepare fresh
before use.
3.4.3 Ascorbyl palmitate standard working solution. Accurately pipette 0.10 mL, 0.20
mL, 0.50 mL, 1.0 mL, and 5.0 mL of the intermediate standard solution, dilute with
citric acid-isoascorbic acid methanol solution, and bring the volume to 10 mL. Mix well
to obtain standard working solutions with mass concentrations of 1.00 μg/mL, 2.00
μg/mL, 5.00 μg/mL, 10.0 μg/mL, and 50.0 μg/mL, respectively. Prepare fresh before
use.
NOTE. The concentration points of the standard working solution can be adjusted appropriately within
the linear range.
3.5 Materials
Organic microporous filter membrane. 0.22 μm.
4 Instruments and equipment
4.1 Liquid chromatograph. It is equipped with a diode array detector or an ultraviolet
detector.
4.2 Electronic balance. The sensitivity is 0.01 mg and 0.1 mg, respectively.
4.3 Tissue grinder.
4.4 Horizontal oscillator.
4.5 Centrifuge. with a speed of not less than 8000 r/min.
5 Analysis steps
5.1 Sample preparation
Liquid samples such as peanut oil and fruit and vegetable juice beverages shall be
shaken well before testing; powdered or paste-like samples with uniform texture, such
as milk powder and fruit puree, shall be thoroughly mixed before testing; other solid or
semi-solid samples, such as canned goods, bread, oatmeal, and candy, shall be
pulverized (for gum-based candies, chopping or freeze-pulverizing may be necessary),
sieved to ensure a particle size of less than 2 mm, and then mixed evenly. Samples shall
be stored in a light-proof, airtight container under cold conditions.
5.2 Sample processing
5.2.1 Sample extraction
Weigh 5 g of the sample (accurate to 0.001 g) and place it in a 50 mL centrifuge tube.
Add 30 mL of citric acid-isoascorbic acid methanol solution, tighten the cap, place the
tube on a horizontal oscillator, and extract for 10 min by oscillating at a rate of not less
than 250 times/min. Transfer the entire sample to a 50 mL volumetric flask, and dilute
to the mark with citric acid-isoascorbic acid methanol solution. Mix well.
5.2.2 Purification
The above sample extract is filtered through filter paper (discarding the initial filtrate)
or centrifuged at 8000 r/min for 1 min to obtain the supernatant, and then filtered
through a 0.22 μm organic microporous membrane into a brown sample vial for analysis
by a high-performance liquid chromatograph.
NOTE. If necessary, the sample solution can be diluted with citric acid-isoascorbic acid methanol
solution to ensure that the concentration of ascorbyl palmitate in the sample solution is within the range
of the standard working solution concentration; the sample solution shall be tested within 12 hours.
5.3 Blank test
Except for the absence of a sample, all other steps are performed simultaneously with
the sample processing as per 5.2.
5.4 Instrument reference conditions
5.4.1 Chromatographic column. C18 column (4.6 mm×150 mm, 5 μm) or a column with
equivalent performance.
5.4.2 Detection wavelength. 245 nm.
5.4.3 Mobile phase. A is a methanol-acetonitrile solution (50+50), B is a phosphoric
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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