The Joint FAO/WHO Expert Committee on Food Additives (JECFA) was established following the recommendation of the first Joint FAO/WHO Conference on Food Additives that the two organizations collect and disseminate information on food additives (Joint FAO/WHO Conference on Food Additives, Report. FAO Nutrition Meetings Report Series, No. 11; WHO Technical Report Series, No. 107, 1956). JECFA was first convened in 1956, and has met annually, with a few exceptions, since that time.
As part of its work, JECFA establishes specifications of identity and purity for food additives. These were originally published in FAO Nutrition Meetings Reports Series (NMRS), the WHO Technical Report Series (TRS) or as FAO Food and Nutrition Papers (FNP). However many of these are now out of print, and the first edition of this Compendium was published in 1992 in order to consolidate all of the then current JECFA specifications into a single publication. Since 1992, separate Addenda to this Compendium have been published which contain both newly established specifications and revisions to earlier specifications. The original Compendium and the succeeding Addenda were all published in the FNP series as number 52. This new Combined Compendium replaces the earlier edition and incorporates all the additions and revisions made since 1992, up to and including those contained in FNP 52 Addendum 13. It is being published as the first document under a new publication series, the FAO JECFA Monographs.
Many additive specifications have been revised two or more times as new information or circumstances require. The specifications contained in this Combined Compendium are the most recent version or revision for each additive. This new Combined Compendium is in four volumes as follows:
This Introduction is intended to replace and update the sections in the IPCS document Principles for the Safety Assessment of Food Additives and Contaminants in Food (Environmental Health Criteria 70 (EHC 70), WHO, 1987) that describe the purpose and function of specifications of identity and purity of food additives. It also incorporates the General Notices section of the Guide to Specifications originally published in Food and Nutrition Paper 5 (FNP 5), revision 2. The remainder of FNP 5 has been extensively revised and updated and is now included in Volume 4.
The term 'specifications' refers to the full set of individual specifications criteria for an additive. Specifications are regarded as one of the outputs of JECFA's risk assessment of additives. The term 'safety evaluation' is used to describe the output of JECFA's assessment of the toxicology and other safety data relating to a food additive.
As previously noted, the specifications published in this Combined Compendium represent the specifications current at the time of publication. These are, however, subject to future review and revision as part of JECFA's continuing work. New and revised specifications are published as soon as practicable after the JECFA meetings when these were considered, and JECFA specifications are also available online on the FAO website.
Specifications for flavouring agents are not included in this Compendium, excepting those few which have an additional technological function as a food additive (e.g. carrier solvent). Flavouring agent specifications will, however, still be available online in a searchable database at the FAO website.
The specifications of identity and purity established by JECFA are intended to ensure that the Committee's safety evaluations apply, with a high degree of confidence, to all food additives manufactured according to those specifications. In setting specifications, the Committee also takes into account the need to encourage good manufacturing practice and to maintain the quality of additives on the market, noting that these considerations may add to the assurance of safety that the Committee is seeking as part of its risk assessment function.
JECFA's specifications are also intended to set manufacturing standards for food additives traded in international commerce either as such, or incorporated in food products. In some cases, JECFA specifications will differ from specifications developed by other national and international organizations. JECFA's safety evaluations can, however, be taken to apply to additives complying with these other specifications, provided that the additives are of equal or higher purity than those required by the JECFA specifications, and provided that the test criteria in the JECFA specifications are met.
The setting of specifications of identity and purity is an essential part of the risk assessment of food additives, and current thinking emphasizes the need for the safety evaluation and the specifications to be seen as joint outputs of the risk assessment process. The safety evaluation of an additive should therefore always be read in conjunction with the specifications of identity and purity that describe the additive. The link between the two parts of the risk assessment process is also emphasized by the fact that the latest safety evaluation is quoted in each of the specifications, together with a reference to the JECFA meetings(s) at which the specifications were elaborated and the safety evaluation carried out.
Formulation of Specifications and Information Requirements
The formulation of satisfactory specifications requires detailed information to be made available to the Committee on the method of manufacture of the additive, including information on raw materials and on its chemical characterization. The Committee requires such information to be provided as part of the total data package whenever an additive is submitted for risk assessment, and all such information will be regarded as suitable for making publicly available unless requested otherwise and agreed by the Secretariat. Those submitting data for a JECFA evaluation are advised to consult existing specifications for further guidance, taking note also of the format of specifications described later in this introduction. They should also seek further advice as needed from the JECFA Secretariat.
Specifications may be revised where there is new information available on methods of manufacture or on the characteristics of the substance, or where changes or revisions in analytical methods are needed. Such changes may also prompt a review of the safety evaluation. Similarly, a review of the specifications may be needed if the safety evaluation is reconsidered.
Additives are mainly defined by a combination of (i) a description of their manufacture, (ii) a minimum requirement for the content of the principal functional component(s) of the additive, and (iii) maximum limits for undesirable impurities. The relative importance of these criteria, however, depends on the nature of the additive. Thus, for example, additives composed largely of single components are mainly defined in terms of their chemical purity, whereas the definition of more complex materials, e.g. natural gums, relies more on a description of the raw materials and the method of manufacture.
In some cases, there may be insufficient information for the Committee to elaborate what it regards as fully acceptable specifications. The Committee may then decide to publish the incomplete specifications, but with a designation of 'Tentative'. In such cases the Committee will also state what additional information is required, and set a date by which this must be provided. The Committee will reconsider the specifications once the necessary information has been received, and if it considers that the information is sufficient, it will remove the 'Tentative' designation. If, however, the information is still deficient, or if no information has been provided by the due date, the specifications will be withdrawn.
JECFA specifications incorporate the methods of analysis that are to be used in testing the individual criteria. Information provided in submissions to JECFA should, therefore, always include details of the analytical methods to be used.
Stability and Fate of Additives in Food
Specifications are intended to apply to the additive as marketed. However, some food additives are designed to perform their function by interacting with components of food as in the case of flour improvers, or, for example by removing undesirable constituents, as in the case of antioxidants. Others may be subject to chemical degradation in food or may interact with food components, sometimes with the production of undesirable reaction products.
In order to ensure that test data are relevant to the way the additive is used in food, the Committee requires information on potential reactivity to be provided as part of submissions for the safety evaluation of additives. This information should include data on (a) the general chemical reactivity of the additive; (b) its stability during storage and reactions in model systems; and (c) the reactions of the additive in food systems. The Committee may also set specific limits on potential degradation products in those cases where the additive may be subject to degradation during storage.
JECFA, CCFAC and the Risk Analysis Framework
The respective roles of JECFA and the Codex Committee on Food Additives and Contaminants (CCFAC) have been extensively discussed in recent years in the context of the Codex risk analysis framework. In this context, JECFA is regarded as the expert risk assessment body on additives, contaminants and natural toxicants in food, with CCFAC fulfilling the corresponding risk management role. Thus, CCFAC endorses maximum use levels only for those additives for which JECFA (i) has established specifications of identity and purity and (ii) has completed a safety assessment or has performed a quantitative risk assessment
CCFAC also makes recommendations to the Codex Alimentarius Commission on the possible adoption of JECFA specifications as Codex Advisory Specifications. National food control authorities use Codex Advisory Specifications for enforcement purposes, and for ensuring that additives in international commerce meet agreed standards. National governments may also draw on Codex Advisory Specifications when developing their own regulatory standards.
Referrals to JECFA and JECFA/CCFAC Interaction
Additives can be referred to JECFA by CCFAC, Codex member nations, FAO or WHO. However, CCFAC provides, as part of its risk management function, the primary forum for handling these referrals, and the JECFA Secretariat works closely with CCFAC in setting the agendas for JECFA meetings. CCFAC also has an important role in ensuring that those requesting JECFA evaluations understand that they bear responsibility for providing the supporting information, and for submitting this by the date set by JECFA in its pre-meeting Call for Data.
Requests to JECFA may be for a full risk assessment of an additive, in which case the information provided will need to include both toxicology and other safety data, as well as the data required to develop specifications. Alternatively, requests may be limited to a reconsideration of information relating to specifications. In such cases the Committee will, however, bear in mind the need to ensure that the specifications still provide the requisite assurance of safety, and will, therefore, only make revisions that it considers will not affect the validity of the safety evaluation.
The current mechanism for considering JECFA's specifications at CCFAC meetings involves the convening of an ad hoc Working Group that meets prior to the plenary CCFAC session. The Working Group, which includes delegates from Codex member states and non-Governmental observers, provides a dedicated forum for the discussion of specifications and other issues relating to the work of JECFA, and the presence of both the JECFA Secretariat and past members of JECFA, attending as part of CCFAC member countries' delegations, provides an opportunity for direct interchange between the risk assessors and risk managers. In this way the Working Group can be seen as fulfilling an important risk communication function in the work of JECFA and CCFAC. The Working Group makes recommendations for consideration at the plenary session of CCFAC, including recommendations on future items to be referred to JECFA and recommendations on which JECFA specifications CCFAC should consider referring to the Codex Alimentarius Commission for possible adoption as Codex Advisory Specifications.
Information submitted to JECFA on the identity and purity of food additives should always include details of the relevant analytical methods. Information on the potential compositional variability of the substance should also be given, together with details of any sampling protocols used to assess this. Insufficient information on analytical methodology is one reason why JECFA may be unable to elaborate suitable specifications, or why it may decide that it is only able to assign a 'Tentative' designation, pending receipt of the further information required.
JECFA specifications incorporate guidance on the analytical techniques that should be used to verify the information. Wherever possible, this is done by reference to Volume 4. Otherwise details of the test procedures are set out in the individual specifications monographs.
As JECFA specifications are elaborated for worldwide use, the Committee prefers to quote methods that require the use of apparatus and equipment that is available in most laboratories, provided that such methods are capable of providing results meeting the requirements of the specifications. Methods involving more recently developed techniques or equipment will not normally be quoted until such techniques are accepted internationally and are generally available at reasonable cost. However, reference to specific methods of analysis should not be taken as precluding the use of other methods, provided that these give results of equivalent accuracy and specificity to those quoted.
Changes to analytical methods are reviewed from time to time as part of JECFA's ongoing work. Changes may also be considered when substances are evaluated for the first time, or when new information becomes available on substances that have been previously considered. Changes in analytical methodology may also prompt further consideration of specifications, for example where these changes reveal the possible presence of previously unsuspected contaminants.
Additives other than enzyme preparations and flavouring agents
JECFA specifications for food additives other than enzymes and flavouring agents normally include the headings listed below, which are given in the order in which they appear in the specifications. Note also that aluminium lakes of colouring matters must comply with the Aluminium Lakes of Colouring Matters - General Specifications included in Volume 1 of the Compendium.
The TITLE includes the name selected for the individual additive. This is the name that, in the view of JECFA, most appropriately identifies the substance or substances specified. It will normally correspond with the name given to the substance that was subject to safety evaluation, although in some cases there may be discrepancies, as when the substance is a member of a group that has been evaluated collectively, e.g. the phosphates.
In proposing names for additives, those submitting information should consult the guidelines for designating names set out in WHO Technical Report Series 1989, No 776, part 2.3.4. These allow the use of non-proprietary names established by international bodies, or used in national legislation, and indicate that in the absence of these, the name may be chosen from existing common or trivial names of the substance. The selected name must, however, be non-proprietary, and should be distinctive enough to enable the substance to be clearly distinguished from other food additives.
The TITLE also indicates those cases where the specifications have been designated as 'Tentative'. This designation is used when the Committee regards the information on the additive as insufficient to elaborate fully acceptable specifications, and in such cases the specifications will include an indication of the further information required.
The TITLE further includes a statement referring to the JECFA meeting at which the specifications were prepared. In cases where the specifications supersede earlier specifications, reference will also be made to these earlier specifications. The statement also includes a reference to the Committee's latest safety evaluation of the substance.
Listed in this section are names, acronyms, and abbreviations under which the substance is widely known, other than those used in the TITLE or in the Chemical name(s) (see below). The Codex Alimentarius International Numbering System (INS) number and the USA FD&C number (for colours) are also included here where applicable. Common or trivial names may also be included as synonyms, but registered trade names are not used.
This section normally includes information on the raw materials used in the manufacture of the additive, together with a brief description of the salient points of the manufacturing method. Proprietary manufacturing information may be excluded provided that this has been agreed with the JECFA Secretariat. For some substances, for example those of natural origin or those containing a number of different components, more detailed information, including manufacturing and purification methods, is given as necessary. The possible presence of other substances included in commercial additive preparations - for example anti-caking agents and antioxidants - may also be noted.
Where an IUPAC or IUBMB name exists for an additive, this is generally included under this heading and listed first among the chemical names, whether or not it is the systematic name or the recommended common name.
CAS number (s)
The Chemical Abstract Service registry number(s) (CAS number(s)) for the major component(s) of the substance is normally given here. CAS numbers for substances that are encompassed by the specifications, e.g. specific isomeric forms of the main component(s), may also be included along with suitable descriptions.
Chem ical formula(e)
The chemical formula(e) of the major component(s) of the additive is given here.
The structural formula(e) corresponding to the major component(s) given under the previous heading is given here, as appropriate.
Formula weights are quoted in JECFA specifications in preference to molecular weights in order to avoid the improper use of the latter form. Formula weights are calculated from values quoted by IUPAC in its Table of Standard Atomic Weights.
A quantitative assay requirement is provided here, where applicable, to indicate the minimum acceptable content, or maximum acceptable content range, of the principal functional component(s) of the additive.
Information on physical appearance and other significant properties, e.g. stability and odour, is provided in this section, and will normally include any special conditions required for the storage and use of the additive. Such information is by its nature descriptive and should not be interpreted as rigidly as, for example, the requirements under the DEFINITION section. For safety reasons, descriptions of the taste of a substance are not included.
Functional uses are included in specifications to indicate the technological functions of the additive as used in foods or in food processing. The stated functional uses are not necessarily an exhaustive list, however, and an additive may have uses other than those listed. The functional uses are intended to conform as far as possible to the harmonized Codex Alimentarius International Numbering System (INS) list of functional uses. Where this is not possible, an unlisted term may be used, and in these cases the new term will be referred for further discussion by the Codex Committee on Food Additives and Contaminants (CCFAC) as part of its consideration of the possible adoption of the specifications as Codex Advisory Specifications.
Identification criteria are generally qualitative and provide part of the means for defining the specified additive. Such criteria typically include solubility in water, solubility in organic solvents, colour reactions, absorption spectra, and pH values.
Where possible, test methods refer to procedures detailed in Volume 4. If the test procedure is not included in the published tests given in Volume 4, it will be included here provided it can be described briefly. Otherwise the details will be set out in the TESTS section under the heading IDENTIFICATION TESTS.
Items relating to the purity of the additive, such as limits on impurities and, where appropriate, criteria for microbiological purity, are included under this heading. Limits for trace impurities and for other parameters relating to purity, for example physical properties, are based on the information available on the manufacturing process at the time the specifications were prepared. Limits are set to be consistent with good manufacturing practice and to help provide an assurance of safety, taking into account the use of the additive.
Where possible, test methods refer to procedures detailed in Volume 4. If the test procedure is not included in the published tests given in Volume 4, it will be included here provided it can be described briefly. Otherwise the details of the test will be set out in the TESTS section under the heading PURITY TESTS.
This section describes in full those test procedures referenced in the CHARACTERISTICS section under the heading IDENTIFICATION. The basic principle behind the analytical method is normally included in the narrative, in addition to details of the apparatus and reagents, the analytical procedure, and the method for calculating results.
This section describes in full those test procedures referenced in the CHARACTERISTICS section under the heading PURITY. The basic principle behind the analytical method is normally included in the narrative, in addition to details of the apparatus and reagents, the analytical procedure, and the method for calculating results. Suppliers of reference standards are named in the text.
METHOD OF ASSAY
The Method of Assay includes a description of the principle of the method, a list of the apparatus and reagents required, details of the analytical procedure, and the method for calculating results. Where possible, these are described by reference to procedures listed in Volume 4. Suppliers of reference standards are named in the text.
Enzyme preparations used in food processing, whether from animal, vegetable or microbial sources, must comply with the General Specifications and Considerations for Enzyme Preparations used in Food Processing found in the next section of this Introduction. In addition, enzyme preparations must meet the specifications criteria contained in the individual specifications monographs These normally include the headings listed below, which are given in the order in which they appear in the specifications.
The TITLE of the specifications monograph includes the name given to the enzyme preparation. This will normally correspond to the name of the enzyme activity or activities that most accurately characterize the preparation. Where appropriate, the source material from which the preparation is derived is also included, so that the name will normally take the form [Principal enzyme activity/activities] from [Systematic IUBMB name of source organism] [Description of genetic modification process where applicable]. The name may or may not be the same as the systematic name recommended by the IUBMB Enzyme Commission Nomenclature Committee.
The TITLE also indicates those cases where the specifications have been designated as 'Tentative'. This designation is used when the Committee regards the information on the enzyme preparation as insufficient to elaborate fully acceptable specifications, and in such cases the specifications will include an indication of the further information required and the date by which this must be submitted.
The TITLE further includes a statement referring to the JECFA meeting at which the specifications were prepared. In cases where the specifications supersede earlier specifications, reference will also be made to these earlier specifications. The statement also includes a reference to the Committee's latest safety evaluation of the substance defined by the specifications.
This section includes names and abbreviations under which the preparation is widely known, other than those used in the TITLE. The INS number is also listed where applicable.
This section identifies the animal, plant or microbial sources used to derive the enzyme preparation. The species, strains or variants, strain numbers and plasmid numbers, if from recognized culture collections/depositories (e.g. ATCC), are also given where appropriate. In cases where the source organism has been derived using recombinant DNA technology, a description of this process, including the identity of the host organism, is included.
Listed in this section are the principal enzyme activities exhibited by the preparation. IUBMB-recommended names are preferred and will generally be listed first, but other names may also be included.
Systematic names and numbers
Where IUBMB systematic names and Enzyme Commission (EC) enzyme numbers exist, these are listed for each active principle. CAS numbers are also given here where these exist.
This section includes a description of the substrates acted on by the enzyme preparation, the reactions catalysed, and the resultant products.
Secondary enzyme activities
Listed here, if appropriate, are minor enzyme activities that may be present in the enzyme preparation.
Information on physical appearance is provided in this section, together with other information such as solubility in water, solubility in organic solvents, and relevant details of the manufacturing process. Information on the diluents, carriers, stabilisers, preservatives and immobilization agents that may be present in commercial products is also included in this section.
This section gives the principal and secondary technological function(s) of the enzyme preparation as used in foods or in food processing.
This gives a statement to the effect that all preparations have to conform to the General Specifications and Considerations for Enzyme Preparations used in Food Processing (see the next section of this Introduction).
The enzyme activities of the active principles are listed here. Also listed here are criteria for trace impurities resulting from, for example, the use of carriers and immobilization agents, other than those noted in the General Specifications and Considerations for Enzyme Preparations used in Food Processing (see the next section of this Introduction).
Where possible, the assay methods refer to procedures detailed in Volume 4. Otherwise the details of the test are set out in the section on TESTS.
Assay methods for the enzyme activities of the active principles listed in the CHARACTERISTICS section under the heading IDENTIFICATION are included here in cases where this cannot be done by reference to Volume 4. These describe the principle of the method, apparatus and reagents required, and give details of the analytical procedure and the method of calculating results. Suppliers of standards for test materials are named in the text.
Specifications for flavouring agents are set out in tabular format. They are not included in the Combined Compendium and, as noted above, may be found in the searchable Flavouring Agents database at the FAO website. The specific items included in flavouring agent specifications are set out below.
|No.||The number allocated by JECFA|
|Name||The name by which the flavouring agent is known in the trade|
|Chemical name||The IUPAC name or a similar, more familiar, name|
Usually restricted to not more than four names
|FEMA||The number allocated by the Flavour and Extract Manufacturers Association of the United States|
The number allocated by the Council of Europe. These will be superseded in due course by the European Commission number
|CAS||Chemical Abstracts Service registry number|
Descriptions of the physical form and odour
|Solubility||Solubility in water and in solvents other than ethanol.|
|Solubility in ethanol||
For interpretation see Volume 4
|Boiling point (°C)||At 760 mm Hg, unless specified otherwise|
|ID Test||Identification test method(s) (IR, NMR, MS)|
|Assay min||Minimum assay value. Where this is <95%, other components are also specified. See also Other requirements|
|Acid value max||Upper limit for acid value|
|Refractive index||At 20°C unless otherwise stated|
|Specific gravity||At 25°C unless otherwise stated|
|Other requirements||May include melting point (mp), content of significant other components, and other significant descriptors|
|JECFA||Contains a reference to the JECFA meeting at which the substance was considered. The abbreviations used are: N = New specifications; R = Specifications revised; T = 'Tentative' specifications (additional information required); S = Existing specifications maintained; N,T = New, 'tentative' specifications ; S,T = Existing, 'tentative' specifications maintained.|
|Data required||Listed in the case of 'Tentative' specifications|
The following general specifications were prepared by the Committee at its sixty-seventh meeting (2006) for publication in FAO JECFA Monographs 3 (2006), superseding the general specifications prepared at the fifty-seventh meeting (1) and published in FAO JECFA Monographs 1 (2). These specifications were originally prepared by the Committee at its twenty-fifth meeting (3) and published in FAO Food and Nutrition Papers No. 19 and No. 31/2 (4,5). Subsequent revisions were made by the Committee at its thirty-fifth meeting and published in FAO Food and Nutrition Paper No. 52 (6). Additional amendments were made at the fifty-first meeting and published in FAO Food and Nutrition Paper No. 52 Add. 6 (7), and at the fifty-third meeting (8) and partially published in FAO Food and Nutrition Paper No. 52 Add. 7 (9).
Classification and Nomenclature of Enzymes
Enzymes are proteins that catalyse chemical reactions. The Enzyme Commission of the International Union of Biochemistry and Molecular Biology (formerly the International Union of Biochemistry) classified enzymes into six main classes: oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases (10). Based on the type of reaction catalysed, enzymes are assigned to one of these classes and given an Enzyme Commission (EC) number, a systematic name, and a common name. Other names are also provided, if available. Enzymes used in food processing are often referred to by their common or traditional names such as protease, amylase, malt, or rennet. For enzymes derived from microorganisms, the name of the source microorganism is usually specified, for example, "a-amylase from Bacillus subtilis." For enzymes derived from microorganisms modified by using recombinant DNA techniques (referred to as recombinant-DNA microorganisms or genetically modified microorganisms), the names of both the enzyme source (donor organism) and the production microorganism are provided, for example, "a-amylase from Bacillus licheniformis expressed in Bacillus subtilis."
Enzymes are used in food processing as enzyme preparations. An enzyme preparation contains an active enzyme (in some instances a blend of two or more enzymes) and intentionally added formulation ingredients such as diluents, stabilizing agents, and preserving agents. The formulation ingredients may include water, salt, sucrose, sorbitol, dextrin, cellulose, or other suitable compounds. Enzyme preparations may also contain constituents of the source organism (i.e. an animal, plant, or microbial material from which an enzyme was isolated) and compounds derived from the manufacturing process, for example, the residues of the fermentation broth. Depending on the application, an enzyme preparation may be formulated as a liquid, semi-liquid or dried product. The colour of an enzyme preparation may vary from colourless to dark brown. Some enzymes are immobilized on solid support materials.
Enzyme preparations usually contain one principal enzyme that catalyses one specific reaction during food processing. For example, α-amylase catalyses the hydrolysis of 1,4-α-D-glucosidic linkages in starch and related polysaccharides. However, some enzyme preparations contain a mixture of enzymes that catalyse two or more different reactions in food. Each principal enzyme present in an enzyme preparation is characterized by its systematic name, common name, and EC number. The activity of each enzyme is measured using an appropriate assay and expressed in defined activity units per weight (or volume) of the preparation.
Enzymes used in food processing are derived from animal, plant, and microbial sources. Animal tissues used for the preparation of enzymes should comply with meat inspection requirements and be handled in accordance with good hygienic practice.
Plant material and microorganisms used in the production of enzyme preparations should not leave any residues harmful to health in the processed finished food under normal conditions of use.
Microbial strains used in the production of enzyme preparations may be native strains or mutant strains derived from native strains by the processes of serial culture and selection or mutagenesis and selection or by the application of recombinant DNA technology. Although nonpathogenic and nontoxigenic microorganisms are normally used in the production of enzymes used in food processing, several fungal species traditionally used as sources of enzymes are known to include strains capable of producing low levels of certain mycotoxins under fermentation conditions conducive to mycotoxin synthesis (11-15). Enzyme preparations derived from such fungal species should not contain toxicologically significant levels of mycotoxins that could be produced by these species.
Microbial production strains should be taxonomically and genetically characterized and identified by a strain number or other designation. The strain identity may be included in individual specifications, if appropriate. The strains should be maintained under conditions that ensure the absence of genetic drift and, when used in the production of enzyme preparations, should be subjected to methods and culture conditions that are applied consistently and reproducibly from batch to batch. Such conditions should prevent the introduction of microorganisms that could be the source of toxic and other undesirable substances. Culture media used for the growth of microbial sources should consist of components that leave no residues harmful to health in the processed finished food under normal conditions of use.
Enzyme preparations should be produced in accordance with good food manufacturing practice and cause no increase in the total microbial count in the treated food over the level considered to be acceptable for the respective food.
Substances used in Processing and Formulation
Substances used in processing and formulation of enzyme preparations should be suitable for their intended uses.
In the case of immobilized enzyme preparations, leakage of active enzymes, support materials, crosslinking agents and/or other substances used in immobilization should be kept within acceptable limits established in the individual specifications.
To distinguish the proportion of the enzyme preparation derived from the source material and manufacturing process from that contributed by intentionally added formulation ingredients, the content of total organic solids (TOS) is calculated as follows:
Lead: Not more than 5 mg/kg
Determine using an atomic absorption spectroscopy/inductively coupled atomic-emission spectroscopy (AAS/ICP-AES) technique appropriate to the specified level. The selection of the sample size and the method of sample preparation may be based on the principles described in the Compendium of Food Additive Specifications, Volume 4.
Salmonella species: absent in 25 g of sample
Total coliforms: not more than 30 per gram
Escherichia coli: absent in 25 g of sample (Determine using procedures described in the section on Microbiological Analyses, Volume 4)
Absent in preparations from microbial sources.
Safety assessment of food enzyme preparations has been addressed in a number of publications and documents. Pariza & Foster (11) proposed a decision tree for determining the safety of microbial enzyme preparations. Pariza & Johnson (16) subsequently updated this decision tree and included information on enzyme preparations derived from recombinant-DNA microorganisms. The Scientific Committee on Food (1 7) issued guidelines for the presentation of data on food enzymes. The document includes a discussion on enzymes from genetically modified organisms including microorganisms, plants, and animals. Several international organizations, government agencies, and expert groups have also published discussion papers or guidelines that address safety assessment of food and food ingredients derived from recombinant-DNA plants and microorganisms (18-28). Certain information in these documents may be applicable to enzyme preparations derived from recombinant sources.
An overall safety assessment of each enzyme preparation intended for use in food processing should be performed. This assessment should include an evaluation of the safety of the production organism, the enzyme component, side activities, the manufacturing process, and the consideration of dietary exposure. Evaluation of the enzyme component should include considerations of its potential to cause an allergic reaction. For enzyme preparations from recombinant-DNA microorganisms, the following should also be considered:
Descriptions and terms applying to the tests and assays in JECFA specifications include the following:
The quantity of the analytical sample to be used is usually indicated in tests and assays. Unless otherwise specified, the quantity used may deviate by 10% from that stated. All quantitative determinations should be conducted on duplicate test portions and in these cases, the amount actually taken should be accurately weighed or measured and the result of the analysis calculated on this exact quantity. When substances are to be "accurately weighed" in a test or assay, the weighing is to be performed in such manner as to limit the error to ±0.1% or less. Quantities smaller than 100 mg should be weighed to the nearest 0.1 mg.
Certain procedures (e.g. chromatographic and spectrophotometric instrumental analyses, and antibiotic and enzyme assays) require the use of analytical reference standards. Where suitable standards are available from recognized international bodies, these are specified. In the absence of international standards, it has been necessary in some cases to specify the use of reference standards available from such organizations as the British Pharmacopoeia (BP), Food Chemicals Codex (FCC), National Formulary (NF) of the United States, or the United States Pharmacopeia (USP). The addresses of these organizations may be found in the individual monographs.
With the exception of volumetric flasks and other exact measuring or weighing devices, directions to use a certain size or type of container or other laboratory apparatus are intended only as recommendations, unless otherwise specified. In certain unavoidable cases, the Committee has found it necessary, for accurate description, to use a proprietary name to indicate a certain type of instrument (e.g. spectrophotometer or chromatograph) that is known to give satisfactory results in a particular analytical procedure. However, such a listing in specifications does not necessarily constitute endorsement of the specified instrument by the Committee, nor does it imply that similar instruments from other sources cannot be used with equal or better satisfaction, or that they are of lesser quality or utility than the instrument named.
The instruction to "perform a blank determination", or similar instructions, indicates that a reagent blank determination should be conducted, in which the same quantities of the same reagents are used, and the same procedure is repeated in every detail, except that the substance being tested is omitted.
A direction that a substance is to be "dried to constant weight" means that the drying should be continued until two consecutive weighings differ by not more than 0.5 mg per g of sample taken, the second weighing to follow an additional hour of drying time at the temperature specified. The direction to "ignite to constant weight" means that the ignition should be continued at a temperature of 450°-550°, unless otherwise specified, until two consecutive weighings do not differ by more than 0.5 mg per g of sample taken, the second weighing to follow an additional 30 min ignition period, depending upon the nature of the sample tested.
Desiccants and Desiccators
The expression "in a desiccator" means the use of a tightly closed container of appropriate size and design in which a low moisture content can be maintained by means of a suitable desiccant. Preferred desiccants include, but are not limited to, anhydrous calcium chloride, magnesium perchlorate, phosphorus pentoxide, and silica gel.
The quantity of an indicator solution used should be 0.2 ml (approximately 3 drops), unless another quantity is specified.
The term "odourless" applies to the examination, after exposure to air for 15 min, of between 1 and 25 g of the substance that has been transferred from the original container to an open evaporating dish of about 100 ml capacity.
Reagents used in tests and assays should be of appropriate analytical grade and should contain no interfering impurities.
Where tolerance limits are expressed numerically, the values are considered to be significant to the number of digits shown. Thus, "not less than 99.0%" means 99.0% but not 99.00%. Values obtained in tests and assays should be rounded off to the nearest indicated digit according to the commonly used practice of rejecting or increasing numbers less than or greater than 5. For example, a requirement of not less than 96.0% would be met by a result of 95.96% but not by a result of 95.94%. When the digit to be dropped is exactly 5, the value should be rounded off to the closest even digit. Thus, both 1.4755 and 1.4765 would be rounded off to 1.476. When a range is stated, the upper and lower limits are inclusive, so that the range consists of the two values themselves, properly rounded off, and all intermediate values between them.
All solutions, unless otherwise specified, are to be prepared with distilled or deionized water. Directions for the preparations of "TS" (test solutions), "TSC" (colorimetric solutions) and "PbT" (lead free solutions) are provided in Volume 4. Where volumetric solutions of definite concentration are directed to be used in quantitative determinations, standardized solutions of other concentrations may be employed, unless otherwise specified, if allowance is made for the calculation factor and if the error of measurement is known not to be increased significantly thereby. Unless otherwise specified, it should be understood that concentrations of solutions prepared from liquids only are expressed in terms of volume in volume (v/v), and solutions of solids in liquids are expressed in terms of weight in volume (w/v). Thus, expressions such as "1 in 10" or "10%" mean that 1 part by volume of a liquid, or 1 part by weight of a solid, is to be dissolved in a volume of the diluent or solvent sufficient to make the finished solutions 10 parts by volume. For other types of solutions (e.g. gases in liquids), and where the above guidelines do not apply, the directions will specify the basis on which the concentration is determined (e.g. w/w, v/w).
Unless otherwise specified, temperatures are expressed in degrees Celsius, and all measurements are to be made at 20°. Ordinary procedures not involving precise instrumental measurements may be conducted at ambient temperature (approximately 15°-30°) unless a particular temperature is specified in a test or assay.
The terms "clear", "almost clear", "very slightly turbid", and "turbid", as specified in Purity Tests for "Clarity and colour of solution", are defined in the individual monographs. The term "no turbidity is produced" means that the clarity of the solution does not change.
The unqualified use of the term "in vacuum" or "in vacuo" means a pressure at least as low as that obtainable by an efficient aspirating water pump (i.e. not higher than about 20 mm of mercury).
The term "water-bath" means a bath of boiling water, unless water at some other temperature is indicated. An alternative form of heating may be employed, provided that the required temperature is approximately maintained and not exceeded.
JECFA generally uses SI units and units accepted for use with SI. The units and abbreviations commonly used in specifications are as follows:
m = metre
cm = centimetre (10-2 m)
mm = millimetre (10-3 m)
μm = micrometre (10-6 m)
nm = nanometre (10-9 m)
g = gram
kg = kilogram (103 g)
mg = milligram (10 -3g)
μg = microgram(10-6 g)
ng = nanogram (10-9 g)
1 = litre
ml = millilitre(10-3l)
ul = microlitre(10-6l)
h = hour(s)*
min = minute(s)*
sec = second(s)
° = degrees Celsius
N = normality (gram equivalents per litre)
M = molarity (moles per litre)
cm-1 = wave number
bar = unit of pressure (kgm-1 sec-2 )*
mm Hg = mm of mercury, unit of pressure **
Rf = ratio of (distance of spot from point of application): (distance moved by solvent)
rpm = revolutions per minute
*Time in minutes or hours, and volume in litres or decimals of litres, are outside the SI, but are accepted for use within it. The term 'bar' is also outside the SI, but is subject to review.
**'mm Hg' is outside the SI system, but is in common use and is retained: 760 mm Hg is equal to 1.013 bar.
JECFA specifications include various abbreviations and acronyms, as set out below.
ADI = Acceptable Daily Intake (expressed in mg/kg bw)
AOAC = AOAC INTERNATIONAL, formerly the Association of Official Analytical Chemists
ASTM = ASTM INTERNATIONAL, formerly the American Society for Testing and Materials
ATCC = American Type Culture Collection
CAC = Codex Alimentarius Commission
CAS = Chemical Abstracts System
C.I. = Colour Index
EC = Enzyme Commission of IUBMB (for systematic nomenclature and numbering system of
FD&C = Food, Drug and Cosmetic
FCC = Food Chemicals Codex (USA)
FNP = FAO Food and Nutrition Paper
FNS = FAO Food and Nutrition Series
FW = Formula weight
G(L)C = Gas (Liquid) chromatography
(HP)LC = (High Performance) Liquid Chromatography
INS = Codex Alimentarius International Numbering System (for food additives)
ISO = International Organization for Standardization
IUBMB = International Union of Biochemistry and Molecular Biology
IUPAC = International Union of Pure and Applied Chemistry
JECFA = Joint FAO/WHO Expert Committee on Food Additives
i.d. = internal diameter
o.d. = outer diameter
IR = Infrared
meq = milliequivalent
MW = Molecular weight
Mol. Wt = Molecular weight
MTDI = Maximum Tolerable Daily Intake
soln = solution
NMRS = FAO Nutrition Meeting Report Series
TLC = Thin Layer Chromatography
TRS = WHO Technical Report Series
UV = Ultraviolet
VIS = Visible wavelength