GB 1886.40-2015 (GB1886.40-2015) & related versions
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National Food Safety Standard -- Food Additives -- L- malic acid
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GB 1886.40-2015
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GB 1886.40-2015
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GB 1886.40-2015
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standard - Food additives - L-malic acid
ISSUED ON: NOVEMBER 13, 2015
IMPLEMENTED ON: MAY 13, 2016
Issued by: National Health and Family Planning Commission of PRC
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Chemical name, molecular formula, structural formula, relative molecular mass ... 4
3 Technical requirements ... 4
Appendix A Testing method ... 6
Appendix B Typical HPLC chromatogram and relative retention time for determination
of fumaric acid or maleic acid content ... 13
National food safety standard - Food additives - L-malic acid
1 Scope
This standard applies to the food additive L-malic acid, which is produced by enzyme
engineering and fermentation methods.
2 Chemical name, molecular formula, structural formula,
relative molecular mass
2.1 Chemical name
L-hydroxysuccinic acid
2.2 Molecular formula
C4H6O5
2.3 Structural formula
2.4 Relative molecular mass
134.09 (according to 2007 international relative atomic mass)
3 Technical requirements
3.1 Sensory requirements
Sensory requirements shall comply with Table 1.
Appendix A
Testing method
A.1 Warning
Some test procedures specified in the test methods may lead to hazardous situations.
Operators shall take appropriate safety and health measures.
A.2 General provisions
Unless other requirements are specified, the reagents and water used in this standard
refer to analytically pure reagents and grade-3 water specified in GB/T 6682. The
standard titration solutions, standard solutions for impurity determination, preparations
and products used in the test, shall be prepared in accordance with the provisions of
GB/T 601, GB/T 602, GB/T 603, unless otherwise specified. The solutions used in the
test refer to aqueous solutions, unless the solvent used to prepare them is specified.
A.3 Identification test
A.3.1 Reagents and materials
A.3.1.1 Ammonia solution: 2 + 3.
A.3.1.2 p-Aminobenzenesulfonic acid solution: 10 g/L.
A.3.1.3 Sodium nitrite solution: 200 g/L.
A.3.1.4 Sodium hydroxide solution: 40 g/L.
A.3.2 Identification method
A.3.2.1 Color development test of ammonium malate salt
Weigh 0.5 g of the specimen, accurate to 0.01 g. Place it in a 50 mL test tube. Add 10
mL of water to dissolve. Use ammonia solution to neutralize it. Add 1 mL of p-
aminobenzene sulfonic acid solution. Heat in a boiling water bath for 5 minutes. Add 5
mL of sodium nitrite solution. Place it in a water bath and heat for 3 minutes. Then add
5 mL of sodium hydroxide solution. The test solution shall immediately turn red.
A.3.2.2 Optical rotation characteristic test
The test method is the same as A.5. The specimen aqueous solution shall have left-
handed characteristics.
A.4 Determination of L-malic acid (C4H6O5) content
A.4.1 Summary of methods
Use phenolphthalein as the indicator. Use the sodium hydroxide standard titration
solution, to titrate the specimen aqueous solution. Based on the consumption of the
sodium hydroxide standard titration solution, calculate the total acid content in C4H6O5
as the L-malic acid content.
A.4.2 Reagents and materials
A.4.2.1 Carbon dioxide-free water.
A.4.2.2 Sodium hydroxide standard titration solution: c (NaOH) = 1.0 mol/L.
A.4.2.3 Phenolphthalein indicator solution: 10 g/L.
A.4.3 Analytical procedures
A.4.3.1 Weigh 2.0 g of specimen, accurate to 0.0002 g. Add 20 mL of carbon dioxide-
free water to dissolve. Add 2 drops of phenolphthalein indicator solution. Use sodium
hydroxide standard solution to make titration, until it is reddish. If it does not fade after
30 s, it is taken as the end point.
A.4.3.2 During the measurement, follow the same steps as the measurement, to do a
blank test using the same amount of reagent solution, without adding a specimen.
A.4.4 Result calculation
The mass fraction w1 of L-malic acid (C4H6O5) is calculated according to formula (A.1):
Where:
V1 - The volume of sodium hydroxide standard titration solution, which is consumed
by the specimen, in milliliters (mL);
V0 - The volume of sodium hydroxide standard titration solution, which is consumed
by the blank test, in milliliters (mL);
1000 - Conversion factor;
c - The concentration of sodium hydroxide standard titration solution, in moles per
liter (mol/L);
M - The molar mass of L-malic acid, in grams per mole (g/mol),
A.7.2.1 Nitric acid solution: 1 + 2.
A.7.2.2 Dextrin solution: 20 g/L.
A.7.2.3 Silver nitrate solution: 20 g/L.
A.7.2.4 Hydrochloric acid standard solution: c (HCl) = 0.1 mol/L.
A.7.2.5 Turbidity standard solution: Containing chlorine (Cl) 0.01 mg/mL. Measure
14.1 mL ± 0.02 mL of c (HCl) = 0.1 mol/L hydrochloric acid standard titration solution.
Place it in a 50 mL volumetric flask. Dilute to the mark. Pipette 10 mL ± 0.02 mL of
this solution into a 1000 mL volumetric flask. Add water to dilute to the mark. Shake
well.
A.7.3 Analytical procedures
Weigh about 1.0 g of the specimen, accurate to 0.01 g. Place it in a colorimetric tube.
Add 20 mL of water to dissolve it. Use it as the specimen solution. Take another
colorimetric tube. Add 0.50 mL of turbidity standard solution accurately. Add water to
20 mL. Add 1 mL of nitric acid solution, 0.2 mL of dextrin solution, 1 mL of silver
nitrate solution. Shake well. Place in the dark for 15 min, as a standard turbidity solution.
In the absence of direct sunlight, if the turbidity of the specimen solution is no greater
than the turbidity of the standard turbidity solution when observed axially and sideways,
the test is deemed to have passed.
A.8 Determination of sulfate (calculated as SO4)
A.8.1 Method summary
In an acidic (hydrochloric acid) solution, the sulfate and barium chloride in the
specimen generate a white precipitate of barium sulfate. Compare it with the standard
turbidity solution. Do a limit test.
A.8.2 Reagents and materials
A.8.2.1 Hydrochloric acid solution: 1 + 4.
A.8.2.2 Barium chloride solution: 250 g/L.
A.8.2.3 Sulfate (SO4) standard solution: 0.1 mg/mL.
A.8.3 Analytical procedures
Weigh 1 g of the specimen, accurate to 0.01 g. Place it in a 50 mL colorimetric tube. At
the same time, pipette 2 mL ± 0.02 mL of sulfate (SO4) standard solution in another 50
mL colorimetric tube. Add 0.5 mL of hydrochloric acid solution and 1 mL of barium
chloride solution to the two colorimetric tubes, respectively. Use water to dilute it to the
mark. Shake well. After leaving it for 10 minutes, if it is observed that the turbidity
produced by the specimen solution is no deeper than the turbidity produced by the
standard turbidity solution, it means the test has passed.
A.9 Determination of chloride (measured as Cl)
A.9.1 Reagents and materials
Chloride (Cl) standard solution: 0.1 mg/mL.
A.9.2 Analytical procedures
Weigh 1.0 g of specimen, accurate to 0.01 g. Measure 0.4 mL of chloride (Cl) standard
solution (containing 0.04 mg of chloride), to prepare a limited standard solution. The
turbidity of the sample shall not be greater than the standard. Others shall be carried out
according to the provisions of GB/T 9729.
A.10 Determination of fumaric acid or maleic acid
A.10.1 Summary of methods
Use high performance liquid chromatography, under selected working conditions, to
separate each component in the sample solution through a chromatographic column.
Use a UV absorption detector to make detection. Use an external standard method to
quantify it. Calculate the fumaric acid or maleic acid in the specimen.
A.10.2 Reagents and materials
A.10.2.1 Sodium hydroxide solution: 20 g/L;
A.10.2.2 Phosphoric acid solution: Pipette 1 mL ± 0.02 mL of phosphoric acid
(excellent grade pure reagent) into a 1000 mL volumetric flask. Add 100 mL of
methanol (HPLC grade reagent) (the amount can be adjusted according to the column
efficiency). Add water to dilute to the mark. Then filter through 0.45 μm filter
membrane.
A.10.2.3 Fumaric acid standard: Purity ≥ 99.0%.
A.10.2.4 Maleic acid standard: Purity ≥ 99.0%.
A.10.3 Instruments and equipment
A.10.3.1 High performance liquid chromatograph
Equipped with UV detector, or other equivalent detector.
A.10.3.2 High pressure pump
Weigh 50 mg of maleic acid standard, accurate to 0.2 mg. Dissolve in an appropriate
amount of water (add a small amount of sodium hydroxide solution if necessary).
Transfer to a 250 mL volumetric flask. Use phosphoric acid solution to dilute it to the
mark. Pipette 1 mL ± 0.02 mL of this solution. Place it in a 100 mL volumetric flask.
Use phosphoric acid solution to dilute to the mark. Shake well. Filter through a 0.45
μm filter membrane. Carry out ultrasonic degassing.
A.10.5.2 Preparation of specimen solution
Weigh 0.2g of the specimen, accurate to 0.0002 g. Place it in a 50 mL volumetric flask.
Use phosphoric acid solution to dilute to the mark. Shake well. Filter through a 0.45
μm filter membrane. Carry out ultrasonic degassing.
A.10.5.3 Determination
Under the reference chromatographic conditions of A.10.4, perform chromatographic
analysis on the specimen solution and standard solution, respectively. Record the peak
area A1 of the substance to be measured in the chromatogram of the specimen solution,
the peak area A2 of fumaric acid or maleic acid in the chromatogram of the standard
solution.
A.10.6 Result calculation
The mass fraction w2 of fumaric acid or maleic acid is calculated according to formula
(A.3):
Where:
A1 - Peak area of the substance to be measured in the chromatogram of the specimen
solution;
m2 - The injection volume of fumaric acid or maleic acid in the standard solution, in
micrograms (μg);
A2 - Peak area of fumaric acid or maleic acid in the chromatogram of standard
solution;
m3 - The injection volume of the specimen, in micrograms (μg).
......
| Standard ID | GB 1886.40-2015 (GB1886.40-2015) | | Description (Translated English) | National Food Safety Standard -- Food Additives -- L- malic acid | | Sector / Industry | National Standard | | Classification of Chinese Standard | X42 | | Classification of International Standard | 67.220.20 | | Word Count Estimation | 12,147 | | Date of Issue | 2015-11-13 | | Date of Implementation | 2016-05-13 | | Older Standard (superseded by this standard) | GB 13737-2008 | | Administrative Organization | National Health and Family Planning Commission of the People Republic of China | | Regulation (derived from) | National Health and Family Planning Commission Announcement No | | Issuing agency(ies) | National Health and Family Planning Commission of the People's Republic of China | | Summary | This standard applies to enzymatic engineering, fermentation of food additives obtained L-malic acid. |
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