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GB 26405-2011: National Food Safety Standard - Food Additive - Lutein
GB 26405-2011
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard - Food Additive - Lutein
ISSUED ON: MARCH 15, 2011
IMPLEMENTED ON: MAY 15, 2011
Issued by: Ministry of Health of the People's Republic of China
Table of Contents
1 Scope ... 3 
2 Molecular formula, structural formula and relative molecular mass ... 3 
3 Technical requirements ... 3 
Appendix A Inspection methods ... 5 
National Food Safety Standard - Food Additive - Lutein
1 Scope
This Standard applies to the food additive lutein, which is made from Tagetes
erecta L. oleoresin through saponification and extraction. Commercial lutein
products can contain edible vegetable oils, dextrins, antioxidants and other
auxiliary materials, which are used for standardization and other purposes.
2 Molecular formula, structural formula and relative
molecular mass
2.1 Molecular formula
C40H56O2
2.2 Structural formula
2.3 Relative molecular mass
568.88 (according to the international relative atomic mass in 2007)
3 Technical requirements
3.1 Sensory requirements: shall meet the requirements of Table 1.
3.2 Physical and chemical indicators: shall meet the requirements of Table
2.
Appendix A 
Inspection methods
A.1 General provisions
The reagents and water that are used in this Standard, when no other
requirements are specified, refer to analytical reagents and grade-III water
which is specified in GB/T 6682-2008. The standard titration solution which are
used in the test, the standard solutions for impurity determination, the
preparations and products, are all prepared in accordance with the provisions
of GB/T 601, GB/T 602, and GB/T 603, when no other requirements are
specified. The solution used in the test, if not indicated which solvent is used,
refers to aqueous solution.
A.2 Identification test
A.2.1 Insoluble in water, extremely-slightly soluble in n-hexane, soluble in
ethanol and chloroform.
A.2.2 In the test for determining the total carotenoid content, the sample
solution has a maximum absorption near 446 nm.
A.2.3 In the test for determining the lutein content, the retention time of the main
peak of the sample solution in the liquid chromatogram shall be the same as
the retention time of lutein in the standard solution.
A.3 Determination of total carotenoids
A.3.1 Reagents and materials
A.3.1.1 Solvent: a mixture of n-hexane, acetone, toluene and absolute ethanol
(10:7:7:6).
A.3.1.2 Absolute ethanol.
A.3.2 Instruments and apparatuses
Ultraviolet-visible spectrophotometer.
A.3.3 Analysis steps
Accurately weigh 0.03 g of the sample, accurate to 0.000 1 g; use the solvent
of A.3.1.1 to dissolve it; transfer to a 100 mL volumetric flask; add the solvent
of A.3.1.1 to fix volume to the mark; shake well. Take 1 mL of this sample
solution in a 100 mL volumetric flask; use absolute ethanol to fix volume to the
A.4.3.1 Chromatographic column: silica gel column, 4.6 mm × 250 mm, particle
size of 3 μm; or other equivalent chromatographic column.
A.4.3.2 Mobile phase: prepare according to the volume ratio, that is, n-hexane:
ethyl acetate = 70: 30; after mixing uniformly, use a 0.45 μm filter membrane
for filtration; perform ultrasonic degassing for later use.
A.4.3.3 Column temperature: room temperature.
A.4.3.4 Mobile phase flow rate: 1.5 mL/min.
A.4.3.5 Injection volume: 10 μL.
A.4.4 Analysis steps
A.4.4.1 Preparation of standard solution
Respectively weigh about 0.01 g of lutein and zeaxanthin standard substances,
accurate to 0.000 1 g; use mobile phase to dissolve; transfer to a 50 mL
volumetric flask; add mobile phase to fix volume to the mark; shake well for later
use.
A.4.4.2 Preparation of sample solution
Weigh 0.025 g ~ 0.027 g of the sample, accurate to 0.000 1 g; use mobile phase
to dissolve it; transfer it into a 100 mL volumetric flask; add mobile phase to fix
volume to the mark; shake well for later use.
A.4.4.3 Determination
Under the reference chromatographic conditions of A.4.3, determine the
standard solution of lutein and zeaxanthin; record the chromatogram. It is
required that the number of theoretical plates shall be at least 5 000 based on
the lutein peak, and the separation between the main peak and the isomer peak
shall be at least 1.5.
Under the reference chromatographic conditions of A.4.3, measure the sample
solution; determine the chemical composition based on the retention time of the
standard; record the chromatogram. Repeat the experiment twice to get the
average peak area values of lutein and zeaxanthin respectively.
A.4.5 Result calculation
The lutein content is calculated as the mass fraction w1 of lutein; the value is
expressed in %, and calculated according to Formula (A.2):
A.5.3.1 Headspace bottle temperature: 80 °C.
A.5.3.2 Quantitative loop temperature: 85 °C.
A.5.3.3 Transmission line temperature: 100 °C.
A.5.3.4 Headspace bottle equilibrium time: 40.0 min.
A.5.3.5 Gas cycle time: 30.0 min.
A.5.3.6 Pressurization time: 0.2 min.
A.5.3.7 Quantitative loop filling time: 0.2 min.
A.5.3.8 Quantitative loop equilibrium time: 0.05 min.
A.5.3.9 Sampling time: 1.0 min.
A.5.3.10 Headspace bottle pressure: 95.15 kPa.
A.5.4 Analysis steps
A.5.4.1 Preparation of control solution
Precisely draw 1.5 μL of n-hexane in a 100 mL volumetric flask; use
dimethylformamide (DMF) to dilute to the mark; shake well; draw 25.0 mL to a
50 mL volumetric flask; use DMF to fix volume. This is the control solution (in
the control solution, the alkane concentration is 0.004 95 mg/mL).
Sensitivity solution preparation: Draw 1.0 mL of the control solution into a 10
mL volumetric flask; use DMF to fix volume; shake well (the concentration of n-
hexane in the sensitivity solution is 0.000 495 mg/mL).
A.5.4.2 Preparation of sample solution
Accurately weigh about 300 mg of the sample; place it in a 10 mL headspace
bottle; add 3.0 mL of DMF to dilute; shake well.
A.5.4.3 System suitability requirements
Take 3.0 mL of the above control solution of A.5.4.1 in a 10 mL headspace bottle;
continuously inject 6 times. The number of theoretical plates of the main peak
shall not be less than 5 000; the peak area RSD shall not be greater than 10.0%;
the signal-to-noise ratio of the main peak in the sensitivity solution shall not be
less than 3.
A.5.5 Result calculation