GB 5009.292-2023 PDF English
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National food safety standard - Determination of 8'-apo-β,ψ-caroten-8'-al in foods
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GB 5009.292-2023: National food safety standard - Determination of 8'-apo-β,ψ-caroten-8'-al 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/GB5009.292-2023
GB
NATIONAL STANDARD OF THE
PEOPLE'S REPUBLIC OF CHINA
National food safety standard - Determination of β-Apo-8'-
carotene aldehyde in food
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 Contents
1 Scope ... 3
2 Principle ... 3
3 Reagents and materials ... 3
4 Instruments and equipment ... 4
5 Analysis steps ... 5
6 Calculation and presentation of results ... 6
7 Precision ... 7
8 Others ... 7
Appendix A Standard solution mass concentration calibration method ... 8
Appendix B Standard solution chromatogram ... 9
National food safety standard - Determination of β-Apo-8'-
carotene aldehyde in food
1 Scope
This standard specifies the liquid chromatography method for the determination of β-
Apo-8'-carotene aldehyde in food.
This standard is applicable to the determination of β-Apo-8'-carotene aldehyde in
flavored fermented milk, processed cheese, frozen drinks, candies, baked goods, semi-
solid compound seasonings, and beverages.
2 Principle
The sample is saponified with potassium hydroxide solution, and the free β-Apo-8'-
carotene aldehyde is extracted with n-hexane. After the extract is concentrated and
reconstituted with acetonitrile, it is separated by high-performance liquid
chromatography, detected with a UV-visible light detector or diode array detector, and
quantified by an external standard method.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are of analytical grade and the
water is first-grade water specified in GB/T 6682.
3.1 Reagents
3.1.1 Acetonitrile (C2H3N): chromatographically pure.
3.1.2 Absolute ethanol (C2H6O).
3.1.3 n-hexane (C6H14).
3.1.4 Potassium hydroxide (KOH).
3.1.5 2,6-Di-tert-butyl-p-cresol (C15H24O, butylated hydroxytoluene, referred to as
BHT).
3.1.6 Formic acid (CH2O2): chromatographically pure.
3.2 Reagent preparation
3.2.1 Potassium hydroxide solution (200 g/L): Weigh 200 g of potassium hydroxide,
add water to dissolve and dilute to 1000 mL.
3.2.2 0.1% BHT-acetonitrile solution: Weigh 0.1 g of BHT, dissolve it in 100 mL
acetonitrile, and mix well.
3.2.3 0.1% formic acid (volume fraction) solution: Pipette 1 mL of formic acid and
dilute to 1 L with water.
3.3 Standard product
β-Apo-8'-carotene aldehyde (C30H40O, CAS number: 1107-26-2): purity is ≥98%, or a
reference material with national authentication and a Reference Material Certificate.
3.4 Preparation of standard solution
3.4.1 β-Apo-8'-carotene aldehyde standard stock solution (10 μg/mL): Accurately
weigh 1 mg (accurate to 0.01 mg) of β-apo-8'-carotene aldehyde standard, dissolve it
with 0.1% BHT-acetonitrile solution, and transfer it to a 100 mL brown volumetric flask;
make the volume up to 100 mL with 0.1% BHT-acetonitrile solution, and mix well;
store it at -18 ℃, and the period of validity is 3 months.
NOTE: β-Apo-8'-carotene aldehyde standard stock solution needs to be calibrated before use. See
Appendix A for specific operations.
3.4.2 β-Apo-8'-carotene aldehyde standard series working solution: Take appropriate
amounts of β-Apo-8'-carotene aldehyde standard stock solution into 10 mL brown
volumetric flasks, add acetonitrile to make the volume up to the mark, and prepare
working solutions with mass concentrations of 0.0500 μg/mL, 0.100 μg/mL, 0.200
μg/mL, 0.500 μg/mL, and 1.00 μg/mL, respectively. Prepare solutions fresh just before
use.
4 Instruments and equipment
4.1 Liquid chromatograph: equipped with a diode array detector or UV-visible light
detector.
4.2 Balance: The sensitivity is 0.01 g and 0.01 mg, respectively.
4.3 Spectrophotometer.
4.4 Constant temperature oscillating water bath device.
4.5 Tissue masher.
4.6 Rotary evaporator.
5.1.3 Blank test
Except for adding no sample, conduct a blank test according to the measurement steps
in 5.1.2.
5.2 Instrument reference conditions
5.2.1 Chromatographic column: C18 column, 150 mm×4.6 mm (i.d.), the particle size
of 5 μm, or a chromatographic column with equivalent performance.
5.2.2 Mobile phase: acetonitrile + 0.1% formic acid solution (95:5, volume ratio).
5.2.3 Flow rate: 1.0 mL/min.
5.2.4 Detection wavelength: 460 nm.
5.2.5 Column temperature: 40 ℃.
5.2.6 Injection volume: 20 μL.
5.3 Preparation of standard curve
Inject the standard series working solutions into the high-performance liquid
chromatograph respectively, and measure the corresponding peak areas. Taking the
mass concentration of β-Apo-8'-carotene aldehyde in the standard series working
solutions as the abscissa, and the peak area of β-Apo-8'-carotene aldehyde as the
ordinate, draw a standard curve. See Appendix B for the chromatogram of the standard
solution of β-Apo-8'-carotene aldehyde.
5.4 Determination of sample solution
The sample solution is injected into the high-performance liquid chromatograph to
obtain the peak area of the object to be measured; it is put into the standard curve to
calculate the mass concentration of β-Apo-8'-carotene aldehyde in the test solution.
Compared with the standard solution, the change range of the retention time of the
compound chromatographic peak in the sample to be tested shall be within ±2.5%.
6 Calculation and presentation of results
The content of β-Apo-8'-carotene aldehyde in the sample is calculated according to
formula (1).
where:
X -- the content of β-Apo-8'-carotene aldehyde in the sample, in grams per kilogram
(g/kg);
ρ -- the mass concentration of β-Apo-8'-carotene aldehyde in the sample solution
obtained from the standard curve, in micrograms per milliliter (μg/mL);
ρ0 -- the mass concentration of β-Apo-8'-carotene aldehyde in the blank test
obtained from the standard curve, in micrograms per milliliter (μg/mL);
V -- the reconstituted volume of the sample, in milliliters (mL);
m -- the sampling amount of the sample, in grams (g);
1000 -- unit conversion factor.
The calculation result is rounded to 2 significant figures.
7 Precision
The absolute difference between two independent determination results obtained under
repeatability conditions shall not exceed 15% of the arithmetic mean.
8 Others
When the sample weight is 2 g, the detection limit is 0.0002 g/kg, and the quantification
limit is 0.0005 g/kg.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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