Nisin preparation
1414-45-5 · E 234 · Nisin · NISIN A · E234 · 234(i) · Nisin · Nisin preparation · Nisiin · Nisiini · Nisin · Nisina · Nisine · Nizin · Nizinas · Nizină · Nizyna · Nizīns · Νισίνη · НизинE number E234INS 234(i)CAS 1414-45-5
Listed as a preservative by Canada, Codex and the US.
Listed as preservatives (CA, US, Codex)
- Canada: Preservative — Health Canada, List of Permitted Preservatives (List 11)
- United States: Antimicrobial agent — FDA, Substances Added to Food: NISIN PREPARATION
- United States: Antimicrobial agents — 21 CFR 184.1538, citing 21 CFR 170.3(o)(2)
- Codex Alimentarius: Preservative — CXS 192-1995 Table One, INS 234(i)
Nisin is an antibacterial peptide made by a bacterium, used in processed cheese, canned foods and some dairy products. It is one of very few antibiotics permitted in food, and is effective against the spore-forming bacteria heat alone may not kill.
Where it is and isn’t permitted
| Jurisdiction | Status | Scope and limits | Citation |
|---|---|---|---|
| Australia and New Zealand | Permitted, with conditions | 13 food categories highest permitted level 250 mg/kg Show every food category
| Food Standards Code, Schedule 15, S15—5and 12 further provisionsretrieved 2026-10-17 |
| Canada | Permitted, with conditions | highest permitted level 30 mg/kg 8 permitted uses Show every permitted use
| Health Canada, List of Permitted Preservatives (List 11)and 7 further provisionsretrieved 2026-10-17 |
| European Union | Permitted, with conditions | 7 food categories highest permitted level 12.5 mg/kg Show every food category
| Regulation (EC) No 1333/2008, Annex IIand 6 further provisionsretrieved 2026-10-18 |
| Great Britain | Permitted, with conditions | 7 food categories highest permitted level 12.5 mg/kg or mg/l Show every food category
| Assimilated Regulation (EC) No 1333/2008and 6 further provisionsretrieved 2026-10-18 |
| Japan | Permitted, with conditions | Designated additive: Appended Table 1, item 286, ナイシン (“Nisin” in the unofficial English translation). …Designated additive: Appended Table 1, item 286, ナイシン (“Nisin” in the unofficial English translation). Standards for use — Specifications and Standards for Foods, Food Additives, etc. (MHW Notification No. 370 of 1959), Part II (Food Additives), F (Standards for use), entry ナイシン: (1) [may be used only in the foods stated] “ナイシンは、穀類及びでん粉を主原料とする洋生菓子、食肉製品、ソース類、卵加工品、チーズ、ドレッシング、ホイップクリーム類(乳脂肪分を主成分とする食品を主要原料として泡立てたものをいう。以下この目において同じ。)、マヨネーズ、みそ及び洋菓子以外の食品に使用してはならない。” (2) [sets maximum levels of use for the foods stated; states an exception to the preceding sentence] “ナイシンの使用量は、ナイシンAを含む抗菌性ポリペプチドとして、食肉製品、チーズ(プロセスチーズを除く。)及びホイップクリーム類にあっては1kgにつき0.0125g以下、ソース類、ドレッシング及びマヨネーズにあっては1kgにつき0.010g以下、プロセスチーズ及び洋菓子にあっては1kgにつき0.00625g以下、卵加工品及びみそにあっては1kgにつき0.0050g以下、穀類及びでん粉を主原料とする洋生菓子にあっては1kgにつき0.0030g以下でなければならない。ただし、特別用途表示の許可又は承認を受けた場合は、この限りではない。” The Japanese is the standard as published. There is no official English translation of the Notification; the English in square brackets is this site’s description of what each sentence does, not a translation of the foods or the levels. Compositional specifications for this additive, in the same Notification, are not imported and are not shown here; nor are the general rules in F 添加物一般, which apply to every additive. | Regulations for Enforcement of the Food Sanitation Act (MHW Ordinance No. 23 of 1948), Appended Table 1retrieved 2026-10-18 |
| South Korea | Permitted, with conditions | Designated food additive: Standards and Specifications for Food Additives, Part III, 1. …Designated food additive: Standards and Specifications for Food Additives, Part III, 1. 일반식품첨가물 (general food additives), 니신 (“Nisin”, the English the Code prints beside it). Main use (주용도) of 니신: 보존료. Standard for use (사용기준) of 니신: (1) [may be used only in the foods listed; sets a limit on the amount used] “니신은 아래의 식품에 한하여 사용하여야 한다. 니신의 사용량은 1. 가공치즈 : 0.250g/kg 이하 2. 두류가공품 : 0.025g/kg 이하” The Korean is the standard as published. There is no official English translation of the current Code (the English version on MFDS’s website is the 2020 edition and has no legal effect); the English in square brackets is this site’s description of what each sentence does, not a translation of the foods or the levels. Compositional specifications (Part IV) are not imported, nor are the general standards in Part II, which apply to every additive. | Standards and Specifications for Food Additives (식품첨가물의 기준 및 규격), MFDS Notification, Part III (Standards for use, by item)retrieved 2026-10-18 |
| United States | Permitted | 21 CFR 184.1538retrieved 2026-10-17 | |
| Codex Alimentarius | Permitted, with conditions | 19 food categories highest permitted level 25 mg/kg Show every food category and its notes
| CXS 192-1995 Table One, INS 234(i)and 18 further provisionsretrieved 2026-09-07 |
How much is permitted
A listing usually limits the quantity as well as the food. Only one of the forms that takes is a figure: quantum satis and good manufacturing practice permit the amount the technological effect requires and no more, which is a limit rather than the absence of one. Figures below are the range across every food category this page is listed in, and categories differ between regulators — see how much is permitted.
| Australia and New Zealand | 10 mg/kg – 250 mg/kg | 4 numeric limit, 9 GMP |
|---|---|---|
| Canada | 3 mg/kg – 30 mg/kg | 8 numeric limit |
| European Union | 3 mg/kg – 13 mg/kg | 7 numeric limit |
| Great Britain | 3 mg/kg – 13 mg/kg | 7 numeric limit |
| Japan | — | 1 no quantity stated |
| South Korea | — | 1 no quantity stated |
| United States | — | 1 no quantity stated |
| Codex Alimentarius | 3 mg/kg – 25 mg/kg | 19 numeric limit |
GRAS notices to the FDA
A GRAS notice is a company’s conclusion that a substance is generally recognised as safe for the use it describes. FDA’s letter responds to that conclusion, for that notifier and that use. It is not a listing in 21 CFR, and it is not counted as a US permission above. The substance and use are as FDA’s inventory words them; this site matched the notice to this page by exact name.
GRN No. 65 · Nisin · Rhodia, Inc.
Intended use: Use on casings for frankfurters and on cooked meat and poultry products as an antimicrobial agent
FDA’s response (2001-04-20): FDA has no questions
What it is made from
Produced by fermenting Lactococcus lactis, traditionally on a milk-based medium, and then concentrated. That fermentation substrate is why nisin preparations are not necessarily suitable for a dairy-free or vegan diet, and it is not usually declared.
- Vegan status depends on the source
- Vegetarian status depends on the source
- May raise a halal question
- May raise a kosher question
This is not an omission. What this additive is made from is not fixed — depending on the manufacturer it may be plant, animal or microbial in origin, nobody is required to say which, and the same product can change source between production runs. Ask the manufacturer if it matters to you; a label cannot answer it.
Compare other additives on the dietary indexes.
- CAS registry number
- 1414-45-5
The two specifications, compared
The European Union sets identity and purity by regulation; JECFA publishes its own through FAO, advisory until a country adopts it. Both documents are below in full. These are the parameters each puts a number on.
| Parameter | European Union | JECFA |
|---|---|---|
| Assay | stated in prose | stated in prose |
| Loss on drying | no more than 3 % | no more than 3 % |
| Lead | no more than 1 mg/kg | no more than 1 mg/kg |
| Arsenic | no more than 1 mg/kg | — |
| Mercury | no more than 1 mg/kg | — |
The two documents state the same figures for every parameter compared here.
How the EU defines it
The legal specification an additive must meet to be sold as this E number in the EU — what it is, how it may be made, and the purity it must reach. From Commission Regulation (EU) No 231/2012.
E 234 Nisin
Nisin consists of several closely related polypeptides produced by strains of Lactococcus lactis subsp. lactis
Assay, identification and purity criteria
Description and assay
- Einecs
- 215-807-5
- Chemical formula
- C₁₄₃H₂₃₀N₄₂O₃₇S₇
- Molecular weight
- 3 354,12
- Assay
- Nisin concentrate contains not less than 900 units per mg in a mixture of non-fat milk solids and a minimum sodium chloride content of 50 %
- Description
- White powder
Purity criteria
- Loss on drying
- Not more than 3 % (102 °C to 103 °C, to constant weight)
- Arsenic
- Not more than 1 mg/kg
- Lead
- Not more than 1 mg/kg
- Mercury
- Not more than 1 mg/kg
How JECFA defines it
The international specification for identity and purity, prepared by the Joint FAO/WHO Expert Committee on Food Additives and published by FAO. It is a different document from any above, written by a different body and revised on its own cycle; it is advisory, and a country adopts it or does not.
INS 234 Nisin A
Nisin A, produced under appropriate fermentation conditions by Lactococcus lactis subsp. lactis strains, is an antimicrobial polypeptide. Nisin A is produced in a sterilized medium of non-fat milk solids or non-milk-based fermentation source, such as yeast extract and carbohydrate solids. The fermentation process is controlled for time and pH, until optimum nisin A production has been achieved. The nisin is then concentrated, recovered and purified from the fermentation medium by various methods, such as sterile injection, membrane filtration, acidification, salting out, ultrafiltration or spray-drying. The purified nisin A is then standardized with sodium chloride to achieve desired activity levels of nisin preparation. Nisin A is stable, at ambient temperatures and when heated, under acidic conditions (up to pH 3). Nisin A is commercially available as nisin preparation, which contains 2.5% w/w nisin A, >50% sodium chloride; the remaining components of the preparation are products of fermentation that include proteins and carbohydrates related to the starting material used for fermentation. The activity of nisin A is measured in International Units (IU). One IU is defined as the amount in micrograms of nisin A required to inhibit the growth of 1 bacterial cell/1 ml of broth of Lactococcus lactis subsp. Cremoris. 1 IU of nisin A is equivalent to 0.025 μg.
Synonyms: INS No. 234
Assay, identification and purity criteria
Description and assay
- Specification history
- Revised specifications prepared at the 99th JECFA (2024) and published in FAO JECFA Monographs 34 (2025), superseding specifications for Nisin prepared at the 77th JECFA (2013). The Committee reaffirmed the ADI of 0-2 mg/kg bw established at the 77th JECFA.
- C.A.S. number
- 1414-45-5
- Chemical formula
- C 143 H 230 O 37 N 42 S 7 (Nisin A)
- Structural formula
- Abu =alpha-aminobutyric acid, Dha=dehydroalanine, Dhb=dehydrobutyrine (Nisin A)
- Formula weight
- 3354.12 (Nisin A)
- Assay
- Not less than 900 IU of nisin per mg (or 22.5 μg/mg)
- DESCRIPTION
- White to light brown micronized powder
- FUNCTIONAL USES
- Antimicrobial preservative
Identification
- Solubility (Vol. 4)
- Soluble in water at pH 2.5, sparingly soluble at pH 5, insoluble at pH>7 and in non-polar solvents
- Differentiation from other antimicrobial substances
- Passes test See description under TESTS
- Nisin A Activity
- The sample shows nisin A activity See description under METHOD OF ASSAY
- Test: Differentiation from other antimicrobial substances
- STABILITY IN ACIDIC CONDITIONS Sample stock solution: Suspend 1 g of sample in 1 liter of 0.02 N hydrochloric acid to give a solution containing 1 000 IU/ml. Sample Preparation: Make a dilution of the Sample stock solution with 0.02 N hydrochloric acid to arrive at a concentration of 50 IU/ml. Boil this solution for 5 min and measure the nisin A activity as directed under ‘Determination of Nisin A Activity’, in METHOD OF ASSAY. The calculated nisin A concentration of the boiled sample should be 100% (+/- 5%) of the assay value indicating no significant loss of activity following this heat treatment. STABILITY IN ALKALINE CONDITIONS Adjust the pH of the unused portion of the boiled nisin A solution from ‘Stability in acidic conditions’ to 11.0 by adding 5 N sodium hydroxide. Heat the solution at 65 °C for 30 min, and then cool. Adjust the pH to 2.0 by adding hydrochloric acid dropwise. Measure the nisin A activity as directed under ‘Determination of Nisin A Activity’ in METHOD OF ASSAY. Record loss of the antimicrobial activity of nisin A following this treatment. Total loss of the antimicrobial activity should be observed following the treatment described. TOLERANCE OF LACTOCOCCUS LACTIS TO HIGH CONCENTRATIONS OF NISIN A Prepare cultures of L. lactis (ATCC 11454, NCIMB 8586) in sterile skim (<1% fat) milk by incubating for 18 h at 30 °C. Prepare one or more flasks containing 100 ml of litmus milk and sterilize at 121 °C for 15 min. Suspend 0.1 g of sample in the sterilized litmus milk, and allow to stand at room temperature for 2 h. Add 0.1 ml of the L. lactis culture, and incubate at 30 °C for 24 h. Record L. lactis growth. L. lactis will grow at this concentration of sample (about 1 000 IU/ml); however, it will not grow in similar concentrations of other antimicrobial substances. (NOTE: This test will not differentiate nisin A from subtilin.)
- Test: Transglucosidase activity
- DETERMINATION OF NISIN A ACTIVITY Principle Nisin A activity, expressed in International Units (IU), refers to the amount of nisin A required to inhibit growth of 1 bacterial cell in 1 ml of broth. 1 IU of nisin A is equivalent to 0.025 μg. Commercial nisin A preparations consist typically of 2.5% w/w of nisin A, sodium chloride and other solids from the fermentation medium. Solution A ( L. lactis subsp. cremoris (ATCC 14365, NCDO 495)): Prepare a subculture daily by transferring one loopful of the test culture to a McCartney bottle of litmus milk and incubating at 30 °C for 24 h (subculture A). Inoculate a suitable quantity of sterile skim milk with 2 percent of subculture A and place it in a water bath at 30 °C for 90 min. Use immediately. Solution B (Standard solution of nisin A): Dissolve 1 g of Nisin A preparation in 1 L of 0.02 N hydrochloric acid and dilute to 1 000 IU/ ml. Dilute a portion of the standard stock in 0.02 N hydrochloric acid to a final concentration of 50 IU/ml. Prepare a nisin A solution containing 5,000 IU/ml (125 micrograms/ml). Dissolve an accurately weighed quantity of standard nisin A in 0.02 N hydrochloric acid. Immediately before use, dilute the solution further with 0.02 N hydrochloric acid to give 50 units/ml (dilution factor 100). Solution C (Unknown sample): Weigh an amount of sample sufficient to ensure that corresponding tubes of the sample and standard series match, i.e., within close limits, so that the nisin A content in the sample and standard are similar. Dilute the sample solution in 0.02 N hydrochloric acid to obtain an approximate concentration of 50 IU per ml (IU/ml). Solution D (Resazurin): Prepare a 0.0125% w/v solution of resazurin in water immediately prior to use. PROCEDURE y Pipet volumes of the 50 IU/ml nùisin standard (Solution B) and unknown sample (Solution C) into two rows of ten 12-ml bacteriological test tubes as depicted in the table below. Pipet volumes of nisin standard and unknown sample Dilution number 1 2 3 4 5 6 7 8 9 10 Solution B (ml) 0.60 0.55 0.50 0.45 0.41 0.38 0.34 0.31 0.28 0.26 Solution C (ml) 0.60 0.55 0.50 0.45 0.41 0.38 0.34 0.31 0.28 0.26 y Add 4.6 ml of the inoculated milk (Solution A) to each by means of an automatic pipetting device. The addition of inoculated milk should be made in turn across each duplicate row of tubes containing the same nominal concentration, and not along each row of ten tubes (e.g., dilution 1 (C and B), dilution 2 (C and B), etc.). y Place the tubes in a water-bath at 30 °C for 15 min, then cool in an ice-water bath while adding 1 ml resazurin solution to each. y Add the resazurin solution in the same order as the addition of inoculated milk, using an automatic pipetting device. Thoroughly mix the contents of the tubes by shaking. Continue incubation at 30 °C in a water bath for a further 3 – 5 min. y Examine the standard and sample tubes under fluorescent light in a black matte-finish cabinet. Compare the sample tube of the highest concentration that shows the first clear difference in colour (i.e., has changed from blue to mauve) with tubes of the standard to find the nearest match in colour. Make further matches at the next two lower concentrations of the sample with the standard. Interpolation of matches may be made at half dilution steps. Obtain three readings of the sample solution and average them. As the standard tubes contain known amounts of nisin A, calculate the concentration of nisin in the sample solution. y If necessary, convert nisin A activity from IU to micrograms of nisin A, using the conversion factor 1 IU = 0.025 micrograms of nisin A. Alternative method Quantification of Nisin A using HPLC y Apparatus and reagents: HPLC equipped with an UV-Vis or Diode Array Detector (DAD); solvents HPLC purity grade. y External HPLC calibration curve and retention time of nisin: Use dilutions of solution B. Filtrate through a syringe filter (0.45μm) and inject into the HPLC. Establish the retention time of nisin and build a calibration curve. Express nisin in mg/kg. y Sample injection: Inject the same aliquot of the unknown samp into the HPLC, measure the area of the corresponding nisin peak. Substitute it in the calibration curve to obtain the corresponding concentration of natamycin in the unknown sample. Adjust the injection volumes and dilutions of the samples to ensure that the response remains within the linear response range of the detector. Column: reversed-phase Jupiter, 5μm, C18, 300A, 250 mm × 4.6 mm or equivalent Flow rate: 1.0 ml/min Detector: 210-260 nm, quantitative at 214 nm Solvents: A (H2O (0.1% Trifluoracetic acid); B (Acetonitrile/H2O / Trifluoracetic acid (90/10/0.1 V/V) Gradient: Time (min) A (%) B (%) 0 78 22 30 43 57 35 0 100 45 78 22 Expected Retention Time of nisin A: 25 minutes (35-37% of acetonitrile)
Purity criteria
- Loss on drying (Vol. 4)
- Not more than 3.0% (105 °C, 2 h)
- Sodium Chloride (Vol. 4)
- Not less than 50%
- LEAD (Vol. 4)
- Not more than 1 mg/kg Determine using a method appropriate to the specified level. The selection of sample size and method of sample preparation may be based on the principles of the methods described in Vol. 4 (under “General methods, metallic impurities”).
- Microbiological criteria (Vol. 4)
- Salmonella species: Absent in 25 g of sample Total coliforms: Less than 30 cfu per gram Escherichia coli : Absent in 25 g of sample
Every name it appears under
20 recorded names. A label may use any of them.
E number
- E234
INS number
- 234(i)
CAS registry number
- 1414-45-5
As it appears in regulation
- Nisin
- Nisin preparation
Other names
- E 234
- Nisin
- NISIN A
The EU re-evaluation programme
Every additive authorised in the EU before 20 January 2009 was put back through EFSA by Commission Regulation (EU) No 257/2010, and this one is named in it. What the Regulation required is below; whether the opinion has since been published is not something this site records.
- Already re-evaluated when the programme began. Listed in Annex I as “Nisin”, against the words “Re-evaluation completed on 26 January 2006”, and therefore not re-evaluated again under it.
Hazard reference values
The two committees set different figures. EFSA's acceptable daily intake is 0.13 mg/kg bw/day; JECFA's is 2 mg/kg bw/day (2024). An acceptable daily intake is a reference value set by a committee, not a threshold above which harm begins. The lower figure is not a finding that the other body was wrong: the two work to different terms of reference, on their own timetables, and sometimes from different data. This site states both and chooses between them nowhere. See where the two committees differ.
| Type | Value | Population | Assessed by | Opinion |
|---|---|---|---|---|
| ADI | 2 mg/kg bw/day | — | JECFA | 2024 |
| ADI | 0.13 mg/kg bw/day | consumers | EFSA | not identified in the dataset |
Evidence and how bodies read it
What assessing bodies concluded
Each conclusion is paraphrased in the body's own framing, from the published opinion linked beneath it. Where bodies differ, both stand; this site does not adjudicate between them or add a view of its own.
EFSA 2017
Considering a new 90-day study, the ANS Panel calculated an ADI of 1 mg nisin A/kg bw/day for nisin (E 234), higher than the previous ADI of 0.13 mg/kg bw. It concluded that overall exposure was below the new ADI and that proposed extensions of use in unripened cheese and heat-treated meat products would not be of safety concern.
Common questions
Assembled from what is above, in the words the questions are usually asked in. Each answer is the same statement the relevant section makes, with the same citations behind it.
- What is E 234?
- E 234 is Nisin preparation. Nisin is an antibacterial peptide made by a bacterium, used in processed cheese, canned foods and some dairy products. It is one of very few antibiotics permitted in food, and is effective against the spore-forming bacteria heat alone may not kill.
- What is Nisin preparation used for in food?
- It is listed as a preservative — the role Canada, Codex and the US assign it. This page shows every function each regulator records, with the citation for each.
- Where is Nisin preparation permitted?
- Of the seven regulators this site reads, Australia and New Zealand, Canada, the EU, Great Britain, Japan, South Korea, the US and the Codex standard permit it. Each permission is for named foods at named levels, not a blanket one; the table on this page gives them with citations.
- Is Nisin preparation vegan?
- It depends on the source, and a label will not tell you. Nisin preparation can be made from plant or animal material, manufacturers are not obliged to say which, and the same product can change between production runs. Anyone who needs to know has to ask the manufacturer.
Sources for this page
- anz retrieved 2026-10-17
- ca retrieved 2026-10-17
- eu retrieved 2026-10-18
- gb retrieved 2026-10-18
- jp retrieved 2026-10-18
- kr retrieved 2026-10-18
- ecfr retrieved 2026-10-17
- gsfa retrieved 2026-09-07
- euspecs retrieved 2026-10-18
- jecfaspecs retrieved 2026-09-23
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