GB 1886.321-2021 (GB1886.321-2021) & related versions
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National food safety standard - Food additives - Thaumatin
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GB 1886.321-2021
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GB 1886.321-2021
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GB 1886.321-2021
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standard - Food additives -
Thaumatin
ISSUED ON: FEBRUARY 22, 2021
IMPLEMENTED ON: AUGUST 22, 2021
Issued by: National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
1 Scope ... 3
2 Relative molecular mass ... 3
3 Technical requirements ... 3
Annex A Inspection methods ... 5
Annex B High performance liquid chromatogram of thaumatin ... 14
National food safety standard Food additives -
Thaumatin
1 Scope
This Standard is applicable to the food additive thaumatin obtained from the
arils of mature fruit of African thaumatococcus daniellii using water extraction
method.
2 Relative molecular mass
Thaumatin I: 22209 (according to the international relative atomic mass in 2018)
Thaumatin II: 22293 (according to the international relative atomic mass in 2018)
3 Technical requirements
3.1 Sensory requirements
The sensory requirements shall meet the requirements of Table 1.
Table 1 – Sensory requirements
3.2 Physical and chemical indicators
The physical and chemical indicators shall meet the requirements of Table 2.
Annex A
Inspection methods
A.1 General
When other requirements are not specified, the reagents and water used in this
Standard refer to analytically-pure reagents and grade 3 water specified in GB/T
6682. All standard solutions used in the test, standard solutions for impurity
determination, preparations and products are prepared in accordance with the
provisions of GB/T 601, GB/T 602, and GB/T 603 when other requirements are
not indicated. The solution used in the test refers to an aqueous solution when
it is not specified which solvent is used to prepare it.
A.2 Identification test
A.2.1 Solubility identification
It is easily soluble in water but insoluble in acetone.
A.2.2 Ninhydrin test
In 5mL of 0.1% sample solution, add 1mL of freshly prepared ninhydrin solution
(200mg of ninhydrin is dissolved in water, and the volume is set to 100mL). Blue
shall appear.
A.2.3 Liquid chromatography test
On the liquid chromatogram of content determination, the retention time of the
main peak of the specimen solution and the main peak of the reference solution
shall be consistent.
A.3 Determination of content (on a dry basis)
A.3.1 Principle of determination
Thaumatin is a protein compound. It has a characteristic absorption peak of
ultraviolet absorption spectrum at 279nm. The specimen is dissolved by water.
Use liquid chromatography to separate. Use UV detector to test. Use external
standard method to quantify.
A.3.2 Reagents and materials
A.3.2.1 Water: Grade 1 water.
A.3.2.2 Ammonium acetate.
Under reference chromatographic conditions, respectively inject a series of
standard solutions and specimen solutions to determine. According to the
external standard method, use a series of standard solutions as a calibration
table. Refer to Annex B for the reference retention time and chromatogram of
each component.
A.3.6 Result calculation
The mass fraction w1 of thaumatin content is calculated according to formula
(A.1).
Where,
c - The concentration of thaumatin in the test solution, in micrograms per
milliliter (μg/mL);
V - The volume of specimen solution, in milliliters (mL);
m - The specimen mass, in milliliters (mL);
1000 - The conversion factor;
f - The dilution times.
The test results are based on the arithmetic mean of the parallel determination
results. The absolute difference between the two independent determination
results obtained under repeatability conditions is not more than 5% of the
arithmetic mean.
A.4 Determination of specific absorption rate
A.4.1 Principle of determination
Perform spectrophotometric detection at the maximum wavelength (typical
wavelength is 279nm).
A.4.2 Reagents and materials
A.4.2.1 Hydrochloric acid.
A.4.2.2 Water (equivalent to grade two water in GB/T 6682).
A.4.3 Instruments and equipment
A.4.3.1 Dual-beam UV/Visible spectrophotometry.
C - 0.05% thaumatin absorption rate;
20 - Dilution multiple of thaumatin specimen solution;
100 - The conversion factor;
W - The moisture content of thaumatin (%);
WCF - The weight correction factor;
WT - The mass of the thaumatin specimen actually used, in grams (g);
0.5 - The mass of the theoretically used thaumatin specimen, in grams (g).
The test results are based on the arithmetic mean of the parallel determination
results. The absolute difference between two independent determination results
obtained under repeatability conditions is not more than 0.2% of the arithmetic
mean.
A.5 Determination of absorbance
A.5.1 Reagents and materials
A.5.1.1 Hydrochloric acid.
A.5.1.2 Water (equivalent to grade two water in GB/T 6682).
A.5.2 Instruments and equipment
A.5.2.1 Dual beam UV/Visible spectrophotometer.
A.5.2.2 Quartz cuvette with a light path of 10mm.
A.5.2.3 50mL volumetric flask.
A.5.2.4 Analytical balance: Resolution is 0.1 mg.
A.5.2.5 Pasteur pipette.
A.5.2.6 pH meter.
A.5.3 Analysis steps
A.5.3.1 Preparation of specimen solution
Accurately weight about 0.5g of specimen in a 50mL volumetric flask. Use the
right amount of pH2.5 blank solution to dissolve. Set volume to 50mL.
A.5.3.2 Determination
A.6.2.3 75mm×10mm disposable test tube.
A.6.2.4 Vortex mixer.
A.6.2.5 Electric heating constant temperature water bath.
A.6.2.6 1mL adjustable pipette.
A.6.2.7 Analytical balance: Resolution is 0.1mg.
A.6.2.8 Pasteur pipette.
A.6.2.9 pH meter.
A.6.3 Analysis steps
A.6.3.1 Preparation of specimen solution
Accurately weigh about 0.2g of specimen in a 100mL volumetric flask. Use the
right amount of pH2.5 blank solution to dissolve. Set volume to 100mL. Take
0.2mL of specimen solution in a tube. Prepare a set of thaumatin specimens: 4
in total. Take 0.2mL of pH 2.5 blank solution to prepare a set of blank solution.
Use straws to respectively add 1.2mL of cysteine-sulfuric acid reagent to the
blank and the specimen. Use a vortex mixer to thoroughly mix well. Place in ice
for 2min. Move to place at room temperature for 3min. Then immerse in the
boiling water for 3min. The blank solution and the specimen solution are cooled
in ice for 5min.
A.6.3.2 Determination
Adjust the wavelength to 412nm. Warm up the spectrophotometer for 10min.
Place the blank solution in the sample cuvette and reference cuvette to zero
the spectrophotometer. Take out the sample cuvette containing the blank
solution. Place the cuvette that contains the specimen solution. Read the
absorbance value (E412). Repeat the test for each set of specimens and take
the average value.
A.6.3.3 Preparation of standard curve
Use 0.2mLof 10mg/mL~100mg/mL glucose solution to test according to the
above methods. Prepare the standard curve.
A.6.4 Result calculation
According to the standard curve, calculate the carbohydrate content w3 (as
glucose) according to formula (A.5).
......
| Standard ID | GB 1886.321-2021 (GB1886.321-2021) | | Description (Translated English) | National food safety standard - Food additives - Thaumatin | | Sector / Industry | National Standard | | Classification of Chinese Standard | X09 | | Word Count Estimation | 11,123 | | Date of Issue | 2021-02-22 | | Date of Implementation | 2021-08-22 | | Regulation (derived from) | National Health Commission Announcement No. 3 of 2021 |
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