GB 1886.174-2024 English PDF (GB1886.174-2024)

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GB 1886.174-2024: National Food Safety Standards--Food Additives--Enzyme Preparations for Food Industry
GB 1886.174-2024
GB
NATIONAL STANDARD OF THE
PEOPLE'S REPUBLIC OF CHINA
National food safety standard - Food additives - Enzyme
preparations for food industry
ISSUED ON: FEBRUARY 8, 2024
IMPLEMENTED ON: AUGUST 8, 2024
Issued by: National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Terms and definitions ... 4
3 Product classification ... 5
4 Technical requirements ... 5
Appendix A List of excipients for enzyme preparations ... 7
Appendix B Reference method for determination of enzyme activity of some enzyme
preparations ... 10
B.1 Reference method for determination of enzyme activity of some enzyme preparations
... 10
B.2 Determination of α-acetolactate decarboxylase activity ... 11
B.3 Determination of pectinase activity ... 19
B.4 Determination of glucoamylase (amyloglucosidase) activity ... 22
B.5 Determination of transglucosidase activity ... 25
National food safety standard - Food additives - Enzyme
preparations for food industry
1 Scope
This standard applies to enzyme preparations for the food industry that are allowed to
be used in National food safety standard - Standards for uses of food additives (GB
2760) and related announcements.
2 Terms and definitions
2.1 enzyme preparations for the food industry
A preparation product with special catalytic activity and used in the food industry,
which is directly extracted from edible or non-edible parts of animals or plants, or
fermented and extracted from microorganisms (including but not limited to bacteria,
actinomycetes, and fungal strains) obtained through traditional selection or genetic
recombination technology, or prepared through further purification, formulation and
other processes (it may contain one or more active enzyme components).
2.2 enzyme activity
An indicator of the activity of an enzyme to catalyze a specific reaction under certain
conditions.
2.3 antibacterial activity
Properties that inhibit or kill microorganisms.
2.4 excipients for enzyme preparations
Food raw materials and food additives added for the purpose of activity preservation,
circulation storage, and standardized use of enzyme preparation products.
2.5 immobilized enzyme preparations
Product dosage forms that immobilize the enzyme on the carrier by physical and (or)
chemical methods and function in an insoluble state during the food production process.
2.6 immobilized carrier
The material that plays the role of immobilizing enzymes in immobilized enzyme
preparations.
3 Product classification
According to product form, it is divided into solid dosage forms (including immobilized
dosage forms) and liquid dosage forms.
4 Technical requirements
4.1 Requirements for raw materials and excipients
4.1.1 The raw materials and excipients used to produce enzyme preparations shall meet
relevant requirements and shall not produce residual contamination harmful to health
in the final food under the specified conditions of use.
4.1.2 Animal tissues used for extracting enzyme preparations shall comply with meat
quarantine requirements.
4.1.3 Plant tissues used for extracting enzyme preparations shall not become moldy.
4.1.4 Microbial production strains shall be taxonomically and/or genetically identified,
and shall comply with the National food safety standard - Standards for uses of food
additives (GB 2760) and related announcements. The preservation methods and
conditions of strains shall ensure stability and reproducibility between fermentation
batches.
4.1.5 For the activity preservation, circulation and storage, and standardized use of
enzyme preparation products, necessary excipients are allowed to be added to
commercial enzyme preparations. Excipients for enzyme preparations shall not play a
functional role in the final food. They are generally used in appropriate amounts
according to production needs, and the varieties and amounts used shall be reduced as
much as possible on the premise of achieving the expected purpose. The list of
excipients allowed to be used in enzyme preparations is shown in Appendix A.
4.1.6 The immobilized carrier used in the immobilized enzyme preparation shall
comply with the National food safety standard - Food contact materials and products
- Standards for uses of additives (GB 9685), National food safety standard - Standards
for uses of food additives (GB 2760) or other provisions on the list of raw materials
allowed to be used in food.
4.2 Physical and chemical indicators
Product enzyme activity shall meet the claims.
B.2.3.3 Reagents and materials
Unless otherwise specified, the reagents used are all of analytical grade, and the water
is the second-grade water that complies with GB/T 6682.
B.2.3.3.1 Acetoin (3-hydroxy-2-butanone) [CH3COCH(OH)CH3, CAS number: 513-
86-0], with a purity of ≥95%; or a standard obtained national certification and awarded
a reference material certificate.
B.2.3.3.2 MES (9.76 g/L)-sodium chloride (35.064 g/L)-polyoxyethylene dodecyl ether
(1.52 mL/L) buffer solution: Respectively weigh 48.80 g of 2-[N-morpholino]
ethanesulfonic acid (MES) and 175.32 g of sodium chloride into a beaker, dissolve it in
about 4.5 L of water, then add 7.60 mL of 15% polyoxyethylene dodecyl ether solution,
and stir evenly; use about 1 mol/L sodium hydroxide solution to adjust the pH to
6.00±0.05. Then transfer it into a 5000 mL volumetric flask, dilute to volume with water,
and stir evenly. The storage period of this solution at normal temperature (15 ℃~20 ℃)
is 1 week.
B.2.3.3.3 α-acetolactate substrate (2.00 mL/L): Take 100 μL of ethyl-2-acetoxy-2-
methylacetoacetic acid into a 50 mL volumetric flask, and add 6.0 mL of about 0.50
mol/L sodium hydroxide solution; after shaking for 20 minutes, add buffer solution to
approximately 40.0 mL; use approximately 1 mol/L hydrochloric acid to adjust the pH
of the solution to 6.00±0.05, and then dilute to volume with buffer solution. The
solution shall be prepared fresh just before use.
B.2.3.3.4 Naphthol (10.0 g/L)/creatine (1.0 g/L) chromogen: Weigh 5.0 g of 1-naphthol
and 0.5 g of creatine respectively into a 500 mL volumetric flask, use about 1 mol/L
sodium hydroxide solution to dissolve and dilute to volume. The solution shall be
prepared fresh just before use. Keep away from light and take an ice bath during
preparation.
WARNING: 1-Naphthol is flammable and toxic. It has irritation to the eyes and
mucous membranes. Poisoning can occur if it is swallowed or absorbed through
the skin.
B.2.3.3.5 Acetoin stock solution (1.000 g/L): Take a certain amount of acetoin in a test
tube and dissolve it in a 37 ℃ thermostat. The test tube is then placed in ice water to
recrystallize the acetoin. After recrystallization, acetoin does not contain acetoin dimers
that will affect the test results and can be used for the preparation of stock solutions;
weigh 0.100 g of acetoin after recrystallization, accurate to 0.0001 g, and transfer it to
a 100 mL volumetric flask, dissolve in water and dilute to volume.
NOTE: Acetoin is unstable to heat and easily absorbs moisture, so it needs to be stored in a desiccator
and refrigerated (2 ℃~6 ℃). If it is found that the material has obvious moisture absorption, it is
recommended to abandon it. The solution has a shelf life of 1 week under refrigeration.
Weigh a certain amount of specimen from the sample, accurate to 0.0005 g, and d...