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GB 1886.78-2016: National Food Safety Standard -- Food Additives -- Lycopene (Synthetic)
GB 1886.78-2016
GB
NATIONAL STANDARD OF THE
PEOPLE?€?S REPUBLIC OF CHINA
National Food Safety Standard -
Food Additives - Lycopene (synthetic)
??????????????? ????????????(??????)
ISSUED ON: AUGUST 31, 2016
IMPLEMENTED ON: JANUARY 1, 2017
Issued by: National Health and Family Planning Commission 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 Method ... 5??
National Food Safety Standard -
Food Additives - Lycopene (synthetic)
1 Scope
This Standard is applicable to the food additive lycopene prepared by Wittig
condensation reaction using synthetic intermediates commonly used in the production
of other carotenoids in foods as the raw materials.
2 Molecular Formula, Structural Formula and Relative
Molecular Mass
2.1 Molecular Formula
C40H56
2.2 Structural Formula
2.3 Relative Molecular Mass
536.87 (in accordance with 2007 international relative atomic mass)
3 Technical Requirements
3.1 Sensory Requirements
Sensory requirements shall comply with the stipulations of Table 1.
Appendix A
Inspection Method
A.1 General Rules
When other requirements are not indicated, the reagents and water used in this
Standard refer to analytically pure reagents and Grade-3 water specified in GB/T 6682.
When other requirements are not indicated, the standard solutions used in the
reagents, and the standard solutions, preparations and products used for impurity
determination shall be prepared in accordance with the stipulations of GB/T 601, GB/T
602 and GB/T 603. When it is not specified which solvent is used for the preparation,
the solutions used in the test refer to aqueous solutions.
A.2 Identification Test
A.2.1 Almost insoluble in water; soluble in organic solvents, for example, chloroform.
A.2.2 Sensitive to heat, light, air and humidity; easily oxidized.
A.3 Determination of Total Lycopene Content and All-trans-lycopene Content
A.3.1 Method summary
Under specified conditions, use high-performance liquid chromatography to determine
the total lycopene content and all-trans-lycopene content.
A.3.2 Reagents and materials
A.3.2.1 N-hexane: chromatographically pure.
A.3.2.2 Tetrahydrofuran stabilized with 0.025% tert-butylhydroquinone.
A.3.2.3 N-ethyl-diisopropylamine: chromatographically pure.
A.3.2.4 Lycopene standard substance: purity ??? 95%.
A.3.3 Instruments and equipment
A.3.3.1 UV / visible spectrophotometer, equipped with 1 cm absorption cell.
A.3.3.2 High-performance liquid chromatography system, equipped with suitable pump,
sample injector, chromatographic column thermostat and integrator.
A.3.3.3 Chromatographic column: 2 connected stainless-steel columns (250.0 mm ???
4.0 mm).
A.3.3.4 Stationary phase: Nucleosil 300-5, 5 ??m.
A.3.3.5 Reference chromatographic conditions:
a) Flow rate: 0.8 mL/min;
b) Sample injection volume: 20 ??L;
c) Column temperature: 20 ??C;
d) Detection wavelength: 470 nm;
e) Mobile phase: 0.15% (volume fraction) N-ethyl-diisopropylamine in N-hexane
solution.
A.3.3.6 Standard solution for high-performance liquid chromatography determination:
accurately weigh-take 5.5 mg ~ 6.5 mg of lycopene standard substance, place it in a
100 mL measuring flask; add 5 mL of tetrahydrofuran stabilized with tert-
butylhydroquinone to dissolve it; use N-hexane to dilute to the scale. It shall be
considered as a standard solution for high-performance liquid chromatography
determination.
A.3.3.7 Standard solution for spectrophotometric determination: accurately weigh-take
5.0 mL of the standard solution (A.3.3.6) for high-performance liquid chromatography
determination; place it in a 100 ml measuring flask; use N-hexane to dilute to the scale.
It shall be considered as a standard solution for spectrophotometric determination.
A.3.3.8 Sample solution: accurately weigh-take 4.5 mg ~ 5.5 mg of sample; place it in
a 100 mL measuring flask; add 5 mL of tetrahydrofuran stabilized with tert-
butylhydroquinone to dissolve it; use N-hexane to dilute to the scale.
A.3.4 Analytical procedures and result calculation
A.3.4.1 Determination of lycopene with spectrophotometry
A.3.4.1.1 Analytical procedures
Use 1 cm absorption cell; take N-hexane as the blank; at the maximum absorption
wavelength (approximately 470 nm), determine the absorbance of the standard
solution (A.3.3.7) used for spectrophotometric determination.
A.3.4.1.2 Result calculation
The concentration cst of lycopene in the standard solution, expressed in (mg/L), shall
be calculated in accordance with Formula (A.1):
A.4.4.1 Preparation of standard solution
Accurately weigh-take 14.5 mg ~ 15.5 mg of Apo-12?€?-lycopene aldehyde reference
substance; place it in a 50 mL measuring flask. Use toluene stabilized with tert-
butylhydroquinone to dissolve and dilute it to the scale. Accurately weigh-take 2 mL;
place it in a 100 mL measuring flask; use toluene stabilized with tert-butylhydroquinone
to dilute to the scale.
A.4.4.2 Preparation of sample solution
Accurately weigh-take 29.0 mg ~ 31.0 mg of sample; place it in a 10 mL measuring
flask. Use toluene stabilized with tert-butylhydroquinone to dissolve and dilute it to the
scale; conduct ultrasonic treatment for 10 min.
A.4.5 Result calculation
The retention time of apo-12?€?-lycopene aldehyde is approximately 14 min; its relative
retention time relative to all-trans-lycopene is 1.6 min. The mass fraction w3 of apo-12?€?-
lycopene aldehyde shall be calculated in accordance with Formula (A.5):
Where,
As---peak area of sample;
Wst---sampling size of standard substance, expressed in (mg);
10---volume of dissolved sample, expressed in (mL);
Ast---peak area of standard substance;
Ws---sampling size, expressed in (mg);
2,500---volume of dissolved reference substance (50 mL) multiplied by the dilution
factor (50).
A.5 Determination of Triphenylphosphine Oxide (TPPO)
A.5.1 Method summary
Under specified conditions, use high-performance liquid chromatography for detection.
A.5.2 Reagents and materials (all reagents shall all be chromatographic grade)
A.5.2.1 N-hexane.
A.5.2.2 Isopropanol.
A.5.2.3 Tetrahydrofuran.
A.5.2.4 Triphenylphosphine oxide (TPPO) (purity ??? 99%).
A.5.3 Instruments and equipment
A.5.3.1 High-performance liquid chromatography system, equipped with suitable pump,
sample injector and integrator.
A.5.3.2 Chromatographic column: stainless-steel column (150.0 mm ??? 4.6 mm).
A.5.3.3 Stationary phase: silica gel base column Supelcosil Lc-Si, 5 ??m or similar
column.
A.5.3.4 Detector: UV detector.
A.5.3.5 Conditions of high-performance liquid chromatography:
a) Column temperature: 20 ??C;
b) Mobile phase: isopropanol: N-hexane (volume ratio 1 : 24);
c) Flow rate: 1.5 mL/min;
d) Sample injection volume: 50 ??L;
e) Detection wavelength: 210 nm;
f) Retention time of TPPO is approximately 8.1 min.
A.5.4 Analytical procedures
Standard solution: accurately weigh-take 10 mg of TPPO standard substance; use
tetrahydrofuran to dissolve it, then, quantitatively transfer it into a 1,000 m volumetric
flask; use tetrahydrofuran to dilute to the scale.
Sample solution: accurately weigh-take 1,000 mg of sample; use tetrahydrofuran to
dissolve it, then, quantitatively transfer it into a 100 mL volumetric flask; use
tetrahydrofuran to dilute to the scale.
A.5.5 Result calculation
Mass fraction w4 of triphenylphosphine oxide (TPPO) shall be calculated in accordance
with Formula (A.6):