GB 1886.231-2023_English: PDF (GB1886.231-2023)
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GB 1886.231-2023 | English | 159 |
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(National Food Safety Standard Food Additive Nisin)
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GB 1886.231-2023
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GB 1886.231-2016 | English | 479 |
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National Food Safety Standard -- Food Additives -- Nisin from Streptococcus Lactis
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GB 1886.231-2016
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Standard ID | GB 1886.231-2023 (GB1886.231-2023) | Description (Translated English) | (National Food Safety Standard Food Additive Sodium Starch Octenyl Succinate) | Sector / Industry | National Standard | Classification of Chinese Standard | X09 | Word Count Estimation | 5,532 | Date of Issue | 2023-09-06 | Date of Implementation | 2024-03-06 | Issuing agency(ies) | National Health Commission of the People's Republic of China, State Administration for Market Regulation | Summary | This standard applies to the food additive octene prepared by esterification of starch and octenyl succinic anhydride, as well as by combining one or more of enzyme treatment, dextrinization, acid treatment, bleaching treatment, and pregelatinization. Sodium starch succinate. | Standard ID | GB 1886.231-2016 (GB1886.231-2016) | Description (Translated English) | National Food Safety Standard -- Food Additives -- Nisin from Streptococcus Lactis | Sector / Industry | National Standard | Classification of Chinese Standard | X09 | Word Count Estimation | 12,152 | Date of Issue | 2016-08-31 | Date of Implementation | 2017-01-01 | Regulation (derived from) | Announcement of the State Administration of Public Health and Family Planning 2016 No.11 |
GB 1886.231-2023. National food safety standard food additive nisin
National Standards of People's Republic of China
National food safety standards
Food additive nisin
Published on 2023-09-06
Implemented on 2024-03-06
National Health Commission of the People's Republic of China
Released by the State Administration for Market Regulation
Preface
This standard replaces GB 1886.231-2016 "National Food Safety Standard Food Additive Nisin".
Compared with GB 1886.231-2016, the main changes in this standard are as follows.
---The detection method of sodium chloride has been modified, and "GB/T 5009.42" has been modified to "'Silver measurement method' in GB 5009.44-2016";
---Modified the titer detection method.
National food safety standards
Food additive nisin
1 Scope
This standard applies to skim milk solids, yeast extract and other nitrogen-containing substances, and carbon-containing substances as the main raw materials, through lactic acid
2 Molecular formula, structural formula and relative molecular mass
2.1 Molecular formula
2.2 Structural formula
NisinA. The 27th amino acid is histidine (His); NisinZ. The 27th amino acid is asparagine (Asn)
2.3 Relative molecular mass
NisinA.3354.35 (according to.2018 international relative atomic mass)
NisinZ.3330.31 (according to.2018 international relative atomic mass)
3 Technical requirements
3.1 Sensory requirements
Sensory requirements should comply with Table 1.
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GB 1886.231-2016
(Food safety national standard - Food additive - Nisin)
National Standards of People's Republic of China
National standards for food safety
Food Additives Nisin
2016-08-31 released
2017-01-01 Implementation
People's Republic of China
National Health and Family Planning Commission released
National standards for food safety
Food Additives Nisin
1 Scope
This standard applies to the Lactococcus lactis (Lactococcus lactissubsplactis) fermentation and extraction of food additives prepared by the milk
Sericin.
2 molecular formula, structural formula and relative molecular mass
2.1 Molecular formula
C143H230O37N42S7 (NisinA)
C141H228O38N41S7 (NisinZ)
2.2 Structural formula
NisnA. amino acid at position 27 is histidine (His); NisnZ. amino acid at position 27 is asparagine (Asn)
2.3 Relative molecular mass
NisinA. 3354.35 (according to.2013 International Relative Atomic Quality)
NisinZ. 3330.31 (according to.2013 International relative atomic mass)
3 technical requirements
3.1 sensory requirements
Sensory requirements shall comply with the requirements of Table 1.
Table 1 sensory requirements
The project requires a test method
Color light brown to milky white
State powder
The amount of the sample evenly placed in the white porcelain dish, in the natural light to observe its color
And state
3.2 Physical and chemical indicators
Physical and chemical indicators should be consistent with the provisions of Table 2.
Table 2 Physical and chemical indicators
Item Index Test Method
Titer/(IU/mg) ≥ 900 A.3 in Appendix A
Dry reduction, w /% ≤ 3.0 GB 5009.3
Sodium chloride, w /% ≥ 50.0 GB/T 5009.42
Lead (Pb)/(mg/kg) ≤ 1.0 GB 5009.75
Note. The commercialization of nisin products should be in line with this standard of nisin as raw material, can add sodium chloride, milk solids and other accessories made of,
The price is consistent with the claim.
3.3 Microbiological indicators
Microbiological indicators should be in accordance with Table 3.
Table 3 Microbiological indicators
Item Index Test Method
Total number of colonies/(CFU/g) ≤ 10 GB 4789.2
Escherichia coli/(MPN/g) < 3.0 GB 4789.3
Escherichia coli/(MPN/g) < 3.0 GB 4789.38
Salmonella should not be detected GB 4789.4
Appendix A
Testing method
A.1 General provisions
This standard, unless otherwise specified, the purity of the reagents used should be analytical grade, the standard titration solution used, the standard solution for the determination of impurities, the preparation
And products should be prepared according to GB/T 601, GB/T 602, GB/T 603, test water should be consistent with GB/T 6682 in the three water regulations
set. The solution used in the test refers to the aqueous solution when it is not specified in the formulation of the solvent.
A.2 Identification test
A.2.1 Reagents and materials
A.2.1.1 hydrochloric acid solution. 0.02 mol/L.
A.2.1.2 Sodium hydroxide solution. 5 mol/L.
A.2.1.3 Skimmed milk. fat content < 1%.
A.2.1.4 Struid milk medium.
A.2.1.5 Detection of bacteria. Lactococcus lactis (ATCC 11454, NCIMB8586).
A.2.2 Analysis steps
A.2.2.1 Preparation of sample stock solution
Take 1g sample, dissolved in 1L hydrochloric acid solution, solution titer of about 1000IU/mL.
A.2.2.2 Stability test for acid solution
The sample stock solution was diluted to about 50 IU/mL with a hydrochloric acid solution to prepare a sample solution. The sample solution was boiled for 5 min at the titer
The Nisin titer of the boiled sample solution. Calculate the potency of Nisin in the boiled sample and calculate the effect at its potency
(100% ± 5%), indicating no significant loss of activity.
A.2.2.3 Stability test of alkali solution
The sample stock solution was diluted to about 50 IU/mL with a hydrochloric acid solution to prepare a sample solution. The sample solution was boiled for 5 min with hydrogen
Sodium sulphate solution to pH 11.0 The solution was heated to 65 ° C for 30 min and cooled. And then dropping hydrochloric acid to adjust the pH to 2.0, with the price determination
The titer of Nisin in the final solution was determined. Almost complete loss of antibacterial activity occurs after treatment according to the above procedure.
A.2.2.4 Identification of Nisin in Different Antimicrobial Substances
A.2.2.4.1 Lactococcus lactis culture. Lactococcus lactis (working strain) (ATCC 11454, NCIMB8586) in sterile skimmed milk
Cultured at 30 ° C for 18 h.
A.2.2.4.2 Preparation of one or more long-necked flasks containing 100 mL of litmus milk medium, sterilized at 121 ° C for 15 min, 0.1 g
The samples were added to the sterile litmus milk medium and mixed at room temperature for 2 h. 0.1 mL of the test bacteria was added and incubated at 30 ° C
24h
A.2.2.4.3 Detection of bacteria (Lactococcus lactis) can be grown in the titer of the sample (about 1000 IU/mL), but not at similar concentrations
He grows in the bacteriostatic substance.
A.3 Determination of potency
A.3.1 Reagents and materials
A.3.1.1 Streptococcus lactide standard (titer. 1 × 106IU/g).
A.3.1.2 Hydrochloric acid solution. 0.02 mol/L.
A.3.1.3 Tween solution. Tween 20. water = 1. 1.
A.3.1.4 medium (S1). tryptone 0.8%; yeast extract 0.5%; glucose 0.5%; sodium chloride 0.5%; 0.2% sodium hydrogen phosphate;
1.2% to 1.5% of the powder, pH 6.8 to 7.0 after sterilization.
A.3.1.5 Detection of bacteria. yellow micrococcus (NCIB8166).
A.3.2 Analysis steps
A.3.2.1 Preparation of test bacteria (NCIB8166) and preparation of bacterial suspension
A.3.2.1.1 Detection of culture of bacteria
A loop test strain (NCIB8166) was taken from the glycerol or lyophilized tube with a sterile inoculation loop, inoculated on a sterile S1 plate,
Separation, pick out full, smooth edge of the colonies, expanded, then S1 in the test tube slope, incubated in the oven at 30 ℃ 24h, into the 2 ℃ ~
5 ℃ refrigerator.
A.3.2.1.2 Preparation of strain suspension
(NCIB8166) in the refrigerator, eluted with sterile saline to prepare a cell suspension at a concentration of 108 CFU/mL,
spare.
A.3.2.2 Preparation of plates
Preparation of S1 medium.200mL (in proportion to the first agar melting, followed by adding the components dissolved, dissolved in disodium hydrogen phosphate after adding), by
121 ℃, 20min sterilization, let cool to 70 ℃ or so, add 4mL Tween 20 solution, shake well, and so cool to 50 ℃ ~ 55 ℃ left
Right, add the amount of prepared strain suspension, so that the final concentration of bacteria in the culture medium is 1.0 × 106/mL, shake, pour into the level
Placed in a sterilized plate, and so on completely solidified, with a diameter of 7mm punch, the number of holes required to play on the plate, carefully digging holes
Within the agar, into the clean bench hair dryer 1.5h ~ 3.0h (hair time according to the size of the humidity in the air, while controlling the indoor temperature of the most
Low, try not to let the test bacteria grow), dry, set 2 ℃ ~ 5 ℃ refrigerator, to the next day to use.
A.3.2.3 Preparation of standard solutions
Accurately weighed nisin standard (accurate to 0.0001g), dissolved in hydrochloric acid solution, the final concentration of 2mg/mL
(2000IU/mL), shake, diluted with hydrochloric acid 300 times, 600 times, that is high, low dose standard solution.
A.3.2.4 Preparation of sample solution
Weigh a certain amount of sample (accurate to 0.0001g), dissolved in hydrochloric acid solution, diluted into high and low dose sample solution, the lactic acid chain
Coccidia content, according to the estimated unit of high and low doses and standard solution roughly the same.
A.3.2.5 Add the solution dropwise
Remove the plate stored in the refrigerator, with a pipette, take 70μL ~ 80μL standard high-dose solution, random drop in the flat hole
In the drop of 6 holes, and then take 70μL ~ 80μL standard low-dose solution, random drop in the same plate with the same high-dose solution of the remaining 6
Hole in the hole.
The sample solution and the standard solution were dropped on the same plate and operated with the standard.
A.3.2.6 Thermostat culture
After the solution in the wells was completely penetrated, the cells were immersed in a 30 ° C incubator for 16 h to 24 h to measure the diameter of the inhibition zone.
A.3.3 Calculation of results
Measure the diameter of the inhibition zone with a caliper, whichever is the average, and calculate the potency according to formula (A.1)
CSH = CBH × k
(XSH XSL) - (XBH XBL)
(XSH XBH) - (XSL XBL) (A.1)
Where.
CSH --- the titer of the sample solution, in units of international units per milligram (IU/mg);
CBH --- titer of the standard solution, in units of international units per milligram (IU/mg);
XSH - diameter of the inhibition zone due to high dose sample solution in millimeters (mm);
XSL --- the diameter of the inhibition zone due to the low dose sample solution in millimeters (mm);
XBH --- diameter of the inhibition zone due to high-dose standard solution in millimeters (mm);
XBL --- the diameter of the inhibition zone due to the low dose standard solution in millimeters (mm);
k - ratio of high dose to low dose concentration.
If the sample estimate is not within the range of 90% to 110% of the measured value, the need to re-estimate the sample titer, retest.
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