JECFA Technical Meeting

983
W H O Te c h n i c a l R e p o r t S e r i e s
983
Evaluation of certain food additives and contaminants

This report represents the conclusions of a Joint FAO/WHO Expert

Committee convened to evaluate the safety of various food additives and
a food contaminant with a view to concluding as to safety concerns and to
preparing specifications for identity and purity.
The first part of the report contains a general discussion of the principles
governing the toxicological evaluation of and assessment of dietary exposure
to food additives. A summary follows of the Committee’s evaluations of
technical, toxicological and dietary exposure data for seven food additives
Evaluation of certain
(advantame; glucoamylase from Trichoderma reesei expressed in Trichoderma
reesei; glycerol ester of gum rosin; glycerol ester of tall oil rosin; glycerol ester
of wood rosin; nisin; and octenyl succinic acid modified gum arabic) and an
food additives and
assessment of dietary exposure to cadmium from cocoa and cocoa products.
Specifications for the following food additives were revised: annatto extracts
(solvent-extracted bixin and solvent-extracted norbixin); Benzoe tonkinensis;
contaminants
food additives containing aluminium and/or silicon; mineral oil (medium
viscosity); modified starches; paprika extract; phosphates (analytical methods
for the determination of phosphorus and revision of specifications); 3-phytase Seventy-seventh report of the
from Aspergillus niger expressed in Aspergillus niger; potassium aluminium Joint FAO/WHO Expert Committee on
silicate; and potassium aluminium silicate–based pearlescent pigments.
WHO Technical Report Series

Food Additives
Annexed to the report are tables summarizing the Committee’s
recommendations for dietary exposures to and toxicological evaluations of
the food additives and contaminant considered.
Food and Agriculture
Organization of
the United Nations
ISBN 978-92-4-120983-0
TRS 983-cover.indd 1 12/16/2013 6:37:00 PM

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WHO Technical Repor t Series
983
food additives and
contaminants
Seventy-seventh report of the
Joint FAO/WHO Expert Committee on
Food Additives
Organization of
the United Nations
WHO Library Cataloguing-in-Publication Data:
Evaluation of certain food additives and contaminants: seventy-seventh report of the Joint
FAO/WHO Expert Committee on Food Additives.
(WHO technical report series ; no. 983)
1.Food additives - analysis. 2.Food additives - toxicity. 3.Food contaminants - analysis.

4.Food contaminants - toxicity. 5.Diet - adverse effects. 6.Risk assessment. I.World Health
Organization. II.Food and Agriculture Organization of the United Nations. III.Joint FAO/WHO
Expert Committee on Food Additives. Meeting (77th: 2013, Rome, Italy). IV.Series.
ISBN 978 92 4 120983 0 (NLM classification: WA 712)
ISSN 0512-3054
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Typeset in India
Printed in Malta
Contents
1. Introduction 1
1.1 Declarations of interests 1
2. General considerations 3
2.1 Modification of the agenda 3
2.2 Report from the Forty-fifth Session of the Codex Committee
on Food Additives (CCFA) 3
2.3 Report from the Seventh Session of the Codex Committee
on Contaminants in Foods (CCCF) 4
2.4 Principles governing the toxicological evaluation of
compounds on the agenda 5
2.5 Food additive specifications 5
2.5.1 Requirements for submission of analytical methods 5
2.5.2 Analytical method for the determination of residual
solvents by headspace gas chromatography 5
2.5.3 Analytical method for the determination of carbon
number at 5% distillation point 6
2.6 GEMS/Food consumption data 6
2.7 FOSCOLLAB 8
3. Specific food additives 11

3.1 Safety evaluations 11
3.1.1 Advantame 11
3.1.2 Glucoamylase from Trichoderma reesei expressed in
Trichoderma reesei 18
3.1.3 Glycerol ester of gum rosin 21
3.1.4 Glycerol ester of tall oil rosin 22
3.1.5 Glycerol ester of wood rosin 23
3.1.6 Nisin 25
3.1.7 Octenyl succinic acid modified gum arabic 30
3.2 Revision of specifications 32
3.2.1 Annatto extracts (solvent-extracted bixin and
solvent-extracted norbixin) 32
3.2.2 Benzoe tonkinensis 33
3.2.3 Food additives containing aluminium and/or silicon 33
3.2.4 Food additives containing phosphates: Analytical
methods for the determination of phosphorus and
revision of specifications 34
3.2.5 Mineral oil (medium viscosity) 35
3.2.6 Modified starches 35
iii
3.2.7 Paprika extract 35
3.2.8 3-Phytase from Aspergillus niger expressed in
Aspergillus niger 36
3.2.9 Potassium aluminium silicate 36
3.2.10 Potassium aluminium silicate–based pearlescent
pigments 37
4. Contaminants 39
4.1 Cadmium: Assessment of exposure from cocoa and cocoa
products 39
5. Future work and requests for data 47
6. Recommendations 51
Acknowledgements 53
References 55
Annex 1 Reports and other documents resulting from previous

meetings of the Joint FAO/WHO Expert Committee
on Food Additives 57
Annex 2 Toxicological and dietary exposure information and

information on specifications 71
Annex 3 GEMS/Food Cluster Diets 2012 75

Seventy-seventh meeting of the Joint FAO/WHO
Expert Committee on Food Additives
Rome, 4–13 June 2013
Members
Dr D. Benford, Food Standards Agency, London, England, United Kingdom
(Vice-Chairperson)
Dr M. DiNovi, Center for Food Safety and Applied Nutrition, Food and Drug
Administration, College Park, MD, United States of America (USA)
Dr D. Folmer, Center for Food Safety and Applied Nutrition, Food and Drug
Administration, College Park, MD, USA
Dr Y. Kawamura, Division of Food Additives, National Institute of Health
Sciences, Tokyo, Japan
Dr Madduri Veerabhadra Rao, Department of the President’s Affairs, Al Ain,
United Arab Emirates
Mrs I. Meyland, Birkerød, Denmark (Chairperson)
Dr U. Mueller, Food Standards Australia New Zealand, Barton, ACT, Australia
(Joint Rapporteur)
Dr J. Schlatter, Zurich, Switzerland
Dr P. Sinhaseni, Community Risk Analysis Research and Development Center,
Bangkok, Thailand
Mrs H. Wallin, Helsinki, Finland (Joint Rapporteur)
Secretariat
Ms J. Baines, Food Standards Australia New Zealand, Canberra, ACT, Australia
(FAO Expert)
Dr G. Brisco, Joint FAO/WHO Food Standards Programme, Food and Agriculture
Organization of the United Nations, Rome, Italy (Codex Secretariat)
Dr A. Bruno, Joint FAO/WHO Food Standards Programme, Food and Agriculture
Organization of the United Nations, Rome, Italy (Codex Secretariat)
Dr S. Cahill, Agriculture and Consumer Protection Department, Food and
Agriculture Organization of the United Nations, Rome, Italy (FAO Secretariat)
Dr R. Cantrill, AOCS, Urbana, IL, USA (FAO Expert)
Dr V. Carolissen, Joint FAO/WHO Food Standards Programme, Food and
Agriculture Organization of the United Nations, Rome, Italy (Codex
Secretariat)
Dr S. Choudhuri, Center for Food Safety and Applied Nutrition, Food and Drug
Administration, College Park, MD, USA (WHO Expert)
v
Mr S.J. Crossley, Food Safety, Food and Agriculture Organization of the United
Nations, Rome, Italy (FAO Joint Secretary)
Dr V. Fattori, Agriculture and Consumer Protection Department, Food and
Agriculture Organization of the United Nations, Rome, Italy (FAO Secretariat)
Dr M. Feeley, Food Directorate, Health Canada, Ottawa, Canada (WHO Expert)
Dr E. Furukawa, Department of Food Safety and Zoonoses, World Health
Organization, Geneva, Switzerland (WHO Secretariat)
Dr Y. Ito, Division of Food Additives, National Institute of Health Sciences, Tokyo,
Japan (FAO Expert)
Dr F. Kayama, School of Medicine, Jichi Medical University, Tochigi, Japan
(WHO Expert)
Mr J. Kim, Department of Food Safety and Zoonoses, World Health Organization,
Geneva, Switzerland (WHO Secretariat)
Professor S. Rath, Department of Analytical Chemistry, University of Campinas,
Campinas, São Paulo, Brazil (FAO Expert)
Ms M. Sheffer, Ottawa, Canada (WHO Editor)
Dr J.R. Srinivasan, Center for Food Safety and Applied Nutrition, Food and Drug
Administration, College Park, MD, USA (FAO Expert)
Professor I. Stankovic, Faculty of Pharmacy, University of Belgrade, Belgrade,
Serbia (FAO Expert)
Dr A. Tritscher, Department of Food Safety and Zoonoses, World Health
Organization, Geneva, Switzerland (WHO Joint Secretary)
Dr T. Umemura, Biological Safety Research Center, National Institute of Health
Sciences, Tokyo, Japan (WHO Expert)
Dr G. Wolterink, National Institute for Public Health and the Environment (RIVM),
Bilthoven, the Netherlands (WHO Expert)
Dr H.J. Yoon, Hazardous Substances Analysis Division, Ministry of Food and
Drug Safety, Seoul, Republic of Korea (WHO Expert)
vi
Monographs containing summaries of relevant data and toxicological evalu-
ations are available from WHO under the title:
Safety evaluation of certain food additives and contaminants.
WHO Food Additives Series, No. 68, 2013.
Specifications are issued separately by FAO under the title:
Compendium of food additive specifications. FAO JECFA Monographs 14,
2013.
vii
1. Introduction
The Joint FAO/WHO Expert Committee on Food Additives (JECFA) met in

Rome, Italy, from 4 to 13 June 2013. The meeting was opened by Dr Ren
Wang, the Assistant Director-General for the Agriculture and Consumer Pro-
tection Department of the Food and Agriculture Organization of the United
Nations (FAO), on behalf of the Directors-General of FAO and the World
Health Organization (WHO).
Dr Wang informed the meeting of the future strategic direction of FAO fol-
lowing a review guided by the Director-General upon taking office in January
2012. He also referred to the forthcoming FAO Conference at which Mem-
ber countries will be asked to endorse the new direction and future priorities.
Dr Wang emphasized that the scientific advice provided by JECFA contrib-
utes to food safety and underpins international standards of the Codex Ali-
mentarius Commission, which is celebrating its fiftieth year. He expressed
his sincere appreciation to the experts for putting their time and expertise at
the service of FAO and WHO.
1.1 Declarations of interests

The Secretariat informed the Committee that all experts participating in the
seventy-seventh meeting had completed declaration of interests forms. A
potential conflict was identified for Dr Yusai Ito. Data submitted for the revi-
sion of specifications on annatto extracts were conducted in his laboratory.
Dr Ito did not participate in the discussion on this matter.
1
2. General considerations
As a result of the recommendations of the first Joint FAO/WHO Conference

on Food Additives, held in September 1955 (1), there have been 76 previous
meetings of the Committee (Annex 1). The present meeting was convened on
the basis of a recommendation made at the seventy-sixth meeting (Annex 1,
reference 211).
The tasks before the Committee were:
— to elaborate further principles for evaluating the safety of food additives
(section 2);
— to review and prepare specifications for certain food additives (section 3
and Annex 2);
— to undertake toxicological evaluations of certain food additives (section 3
and Annex 2);
— to undertake a dietary exposure assessment of one food contaminant
(section 4 and Annex 2).
2.1 Modification of the agenda

The agenda was modified to include 3-phytase from Aspergillus niger
expressed in Aspergillus niger for modification of the method that is listed
in the specifications, following a request from the Forty-fifth Session of the
Codex Committee on Food Additives (CCFA) (2).
2.2 eport from the Forty-fifth Session of the Codex Committee on

R
Food Additives (CCFA)
The Codex Secretariat provided the Committee with an update of the work of
CCFA since the seventy-sixth meeting of JECFA (Annex 1, reference 211),
including the main achievements and outputs of the Forty-fifth Session of
CCFA (2).
Following the outcome of the seventy-sixth meeting of JECFA, the Forty-fifth
Session of CCFA invited Member countries to submit to JECFA (i) data on actual
use levels for magnesium-containing food additives and for phosphate‑containing
3
food additives; and (ii) new information on the toxicological effects of phosphate
salts, expressed as phosphorus. It also recommended the inclusion of the three
enzymes evaluated by the seventy-sixth meeting of JECFA in the database on pro-
cessing aids, prepared by China; and agreed to revoke the specifications of min-
eral oil (medium and low viscosity) classes II and III (International Numbering
System [INS] 905 e,f,g) and to revise the name and adopt the new specifications
of INS 905e mineral oil, medium viscosity.
The Forty-fifth Session of CCFA finalized work on more than 600 provi-
sions of the Codex General Standard for Food Additives (GSFA) (3) and
concluded the revision of the provisions of aluminium-containing food addi-
tives, as recommended by the seventy-fourth meeting of JECFA (Annex 1,
reference 205). In addition, CCFA recommended the adoption of new and
revised specifications for the identity and purity of 8 food additives and 93
flavourings, prepared by the seventy-sixth meeting of JECFA; and amend-
ments to the INS. With regard to ammonium aluminium silicate, pearlescent
pigment, CCFA agreed to consider its inclusion in the INS in light of the
outcome of the current meeting of JECFA.
The Forty-fifth Session of CCFA agreed on a revised priority list of com-
pounds for evaluation (or re-evaluation) by JECFA, which includes 19 food
additives and 124 flavourings. Because of the large number of substances on
the priority list, CCFA assigned high priority to 10 of them. CCFA continued
its discussion on the prioritized list of 107 food colours evaluated by JECFA
and agreed to consider at its next session a document identifying different
options for the use of the outcomes of the prioritization exercise and other
feasible steps to identify compounds for re-evaluation by JECFA.
The Forty-fifth Session of CCFA further agreed to consider at its next session
a document regarding the inclusion of secondary additives in specifications
and the need to develop guidance on how to address their use and to start
work on the revision of the Codex Guidelines for Simple Evaluation of Food
Additive Intake (4), which aims to assist Member countries, especially devel-
oping countries, in their assessment of dietary exposure to food additives by
reflecting current procedures in place to carry out such work in a simple way.
2.3 eport from the Seventh Session of the Codex Committee on

R
Contaminants in Foods (CCCF)
The Codex Secretariat informed the Committee about the status of work on
cadmium contamination in foods in CCCF since the last re-evaluation of
cadmium by JECFA at its seventy-third meeting (Annex 1, reference 202).
Based on the outcome of this evaluation, the Fifth Session of CCCF (5) agreed
that there was no need to revise the maximum levels (MLs) for cadmium
in the Codex General Standard for Contaminants and Toxins in Food and
4
Feed (6) or the provisions in the Code of Practice Concerning Source Directed
Measures to Reduce Contamination of Food with Chemicals (7).
The Sixth Session of CCCF (8) considered a request for the establishment
of MLs for cadmium in cocoa and cocoa products and decided to include
cadmium on the priority list for evaluation by JECFA for an assessment of
exposure from cocoa and cocoa products. The outcome of the assessment
will be considered by the Eighth Session of CCCF (in 2014) to decide upon
new work on the establishment of MLs for cadmium in cocoa and cocoa
products or any other appropriate risk management options available.
2.4 rinciples governing the toxicological evaluation of compounds

P
on the agenda
In making recommendations on the safety of food additives, the Committee
took into consideration the principles established and contained in the publi-
cation, Environmental Health Criteria, No. 240, Principles and Methods for
the Risk Assessment of Chemicals in Food, published in 2009 (9).
2.5 Food additive specifications
2.5.1 Requirements for submission of analytical methods
The Committee, while encouraging the use of analytical methods published

in Volume 4 of the Combined Compendium of Food Additive Specifications
(Annex 1, reference 180), noted that reference to specific methods of analy-
sis should not be taken as precluding the use of other methods, provided
these methods give results of equivalent accuracy and specificity to those
quoted. In order to assess and ensure the accuracy and reliability of the data
submitted, the Committee recommends the use of methods that are appro-
priately validated. It also recommends that in relevant cases, the detailed
analytical method be provided, together with validation data, in response to
specific JECFA calls for data.
2.5.2 Analytical method for the determination of residual solvents by

headspace gas chromatography
The Committee at its current meeting noted that the specifications for
paprika extract and annatto extracts (solvent-extracted bixin and norbixin)
contained the requirement for the determination of residual solvents. The
use of the general method for residual solvent by headspace gas chromatog-
raphy published in Volume 4 of the Combined Compendium of Food Addi-
tive Specifications (Annex 1, reference 180) was not suitable for the analysis
of these substances because of their low solubility in water or methanol,
listed as the solvents in the general method. The Committee agreed that
alternative solvents were appropriate for the assay of these substances. The
5
Committee noted that there may be many substances that are insufficiently
soluble in water or methanol or may contain methanol as a residual solvent
from the manufacturing process. Therefore, the Committee recommended
that the issue of the suitability of dissolution solvents for the determination
of residual solvents in food additives be investigated at a future meeting.
2.5.3 Analytical method for the determination of carbon number at 5%

distillation point
The Committee at its current meeting noted that the specifications for min-
eral oil (medium viscosity) contained a gas chromatographic method for
the determination of carbon number at 5% distillation point that uses a
packed column. The Committee noted that there was a general requirement
to replace these methods with methods using capillary columns. A method
using a wide-bore column was included in the revised specifications. The
Committee also noted that the original method was included in Volume 4 of
the Combined Compendium of Food Additive Specifications (Annex 1, refer-
ence 180) and recommended that a note be included in Volume 4 to indicate
the availability of a newer method. The Committee further recommended
that the suitability of this method for use in the analysis of similar substances
be evaluated at a future meeting.
2.6 GEMS/Food consumption data

A WHO representative gave a presentation on the new Global Environment
Monitoring System – Food Contamination Monitoring and Assessment Pro-
gramme (GEMS/Food) Cluster Diets to the Committee for consideration and
discussion.
GEMS/Food diets have always reported per capita amounts of food (grams
per person per day) available for consumption, derived from FAO Food Bal-
ance Sheet data, to give the best coverage for all countries in the world, as
dietary information at the individual level from national nutrition surveys is
not available for many countries. Cluster Diets were first developed in 1997
by grouping together countries that had geographical proximity as well as
similar per capita data for 20 key foods. This resulted in 13 clusters of coun-
tries. The 13 Cluster Diets were revised in 2006 using updated FAO Food
Balance Sheet data.
In 2012, a more sophisticated statistical approach was taken, whereby an
expanded set of FAO Food Balance Sheet data for 415 primary or semi-
processed food products for 179 countries for the period from 2002 to 2007
(data available on the FAO web site1) was used to make a substructure of 30
1
http://faostat3.fao.org/home/index.html
6
consumption systems. Clusters of countries were then derived according to
their consumption profiles, which could include one or more consumption
systems (these could be a single food, such as cassava, cows’ milk or rice,
or groups of foods consumed together, such as yams and taro). As a con-
sequence, the new Cluster Diets published by WHO in 2012 report on per
capita food data for 17 clusters and replace the 13 Cluster Diets previously
used (refer to Annex 3 for the grouping of countries in the 17 clusters). Data
in these new Cluster Diets are available at three levels of detail: Level 1 for
17 major food groups (e.g. cereals and cereal products); Level 2 for 62 major
food subgroups (e.g. cereal grains and flours); and Level 3 for 415 precise
food subgroups (e.g. rice). Level 1 and Level 2 data are available on the
WHO web site2.
The Cluster Diets were considered to be most useful for application in inter-
national dietary exposure assessments where the chemical use or contamina-
tion level was likely to be similar worldwide—for example, for an additive
used in beer. In that case, the type and amount of beverages available for con-
sumption would be the main driver of differences in estimated mean popula-
tion chronic dietary exposure to the food chemical across different clusters.
For contaminants, if sufficient concentration data were available for foods at
the individual country level, it would be preferable in most cases to derive
summary concentration data for different foods for each cluster of countries,
which could then be combined with the per capita consumption data to better
estimate mean population chronic dietary exposure to the contaminant for
each cluster.
In addition to Cluster Diets, which represent per capita food data, FAO and
WHO have recently developed a database that collates food consumption
data derived from individual records for 26 countries for use in chronic and
acute dietary exposure assessments. The database was presented to the Com-
mittee, which noted that the data could be filtered by age class, country, food
group (Level 2) and food item (Level 3), allowing users to focus on a specific
data set, depending on the food chemical under consideration.
WHO also presented a study that aimed to determine the validity of the
assumption that per capita food amounts (food available for consumption)
would always be higher than the population mean amount of these foods
derived from individual records from national nutrition surveys. The study
compared information for four food groups from individual chronic food
consumption data for 20 countries (for which data derived from individual
records from surveys of 2 days’ duration or more were available for adults
or the general population) and per capita food data from the seven related
2
http://www.who.int/entity/foodsafety/chem/Cluster_diets_2012_consumption.xls
7
Cluster Diets. Results from the study indicate that Cluster Diets do not
systematically overestimate mean population food consumption amounts,
as had been previously assumed (9). For example, for some processed or
semi-processed foods (e.g. cereal products), the mean population food con-
sumption amounts derived from national survey data were higher than the
corresponding per capita amount from the relevant Cluster Diet. It was noted
that direct comparisons between the two different types of data sets should
be made with caution. In general, the overall sum of raw, semi-processed
and processed food amounts for each food category at Level 2 tended to be
similar in the relevant Cluster Diet and national food consumption data sets,
indicating that there may be some discrepancies in the way in which raw
versus processed foods had been classified in the different types of data sets.
The Committee welcomed the update of the WHO GEMS/Food Cluster Diets
and the establishment of the food consumption database based on individ-
ual records from national surveys. The Committee applied the new Cluster
Diets to evaluations at the present meeting, where relevant. The Committee
noted that both the Cluster Diets and the available individual food consump-
tion data for individual countries should be considered in international food
chemical dietary exposure assessments, with expert judgement required to
determine their appropriate use.
2.7 FOSCOLLAB
The WHO Secretariat presented to the Committee a global platform for food
safety data and information called FOSCOLLAB (Food Safety Collabora-
tion), which was recently launched on the WHO web site3. FOSCOLLAB
enables users to access integrated data and information from the agriculture,
food and human health areas to support decision-making in food safety.
FOSCOLLAB integrates data and information from various existing WHO
databases, such as JECFA evaluations, GEMS/Food contaminant occurrence
data (including level of detection and average concentration by commodity),
GEMS/Food consumption data and WHO Collaborating Centres (including
institutions working in the area of food contaminants), as well as the Codex
General Standard for Contaminants and Toxins in Food and Feed (6). FOS-
COLLAB has the capability to generate reports, which the user can export
into specific formats, such as PDFs or slide presentations. FOSCOLLAB
is built on a modular basis; it currently contains data on food contaminants
and can be expanded in the future with information on other food chemicals,
such as food additives and residues of pesticides and veterinary drugs. Com-
mon denominators between databases are the food category, hazard, country
and year.
3
http://www.who.int/foodsafety/foscollab/en/index.html
8
The WHO Secretariat invited the meeting participants to test FOSCOLLAB
and provide comments for further improvements and to shape its scope and
functionalities, including recommending possible data sets for future inclu-
sion. The Committee appreciated this new tool and is looking forward to
further developments.
9
3. Specific food additives
The Committee evaluated two food additives for the first time and re-evaluated
five others. Seventeen food additives were considered for revision of specifica-
tions only. Information on the safety evaluations and specifications is summa-
rized in Annex 2. Details on further toxicological studies and other information
required for certain substances are summarized in section 5.
3.1 Safety evaluations
3.1.1 Advantame
Explanation
Advantame (N-[N-[3-(3-hydroxy-4-methoxyphenyl) propyl]-L-α-aspartyl]-
L-phenylalanine-1-methyl ester, monohydrate, Chemical Abstracts Service
No. 714229-20-6) is an N-substituted (aspartic acid portion) derivative of
aspartame that is structurally similar to another N-substituted aspartame
derivative, neotame.
Advantame has not previously been evaluated by the Committee. Although
it was submitted at the seventy-sixth meeting (Annex 1, reference 211) for
consideration as a flavouring agent, the Committee decided that it would
be inappropriate to evaluate this substance as a flavouring agent because
it is a high-intensity sweetener, and evaluation as a food additive had been
requested by the Forty-fourth Session of CCFA (10).
Chemical and technical considerations

Advantame appears as a white to yellow powder that is very slightly soluble
in water and sparingly soluble in ethanol. It is used as a high-intensity, non-
nutritive sweetener in tabletop sweeteners and in a wide variety of foods,
and it has been demonstrated to be approximately 100 times sweeter than
aspartame and approximately 20 000–37 000 times sweeter than sucrose.
Advantame is manufactured by N‑alkylation of the aspartic acid portion of
aspartame (L-α-aspartyl-L-phenylalanine methylester), with 3-(3-hydroxy-
4-methoxyphenyl) propionaldehyde produced by selective catalytic
11
hydrogenation from 3-hydroxy-4-methoxycinnamaldehyde. Methanol and
ethyl acetate are used as reaction solvents and recrystallization solvents in
the preparation of advantame.
The final advantame product has a purity of not less than 97.0% on an anhy-
drous basis. Specifications of not more than 1.0% and 1.5% were set for
the advantame acid (N-[N-[3-(3-hydroxy-4-methoxyphenyl) propyl]-L-α-
aspartyl]-L-phenylalanine) and other related substances, respectively. In
addition, the limits for residual solvents, residues on ignition and lead were
specified.
Advantame slowly degrades under acidic conditions and at high temperature
under baking conditions. The main degradation product is the advantame
acid. In contrast to aspartame, advantame does not form the diketopiperazine
derivative, as there is no free amino group to start the internal reaction of
cyclization.
Toxicological data
A comprehensive range of studies on pharmacokinetics and toxicokinetics,
acute toxicity, short-term and long-term toxicity, carcinogenicity, genotoxic-
ity, and developmental and reproductive toxicity undertaken with appropri-
ate standards for study protocol and conduct were taken into consideration in
the safety assessment of advantame.
In all species studied (mice, rats, rabbits, dogs and humans), advantame is
rapidly converted to advantame acid (de-esterified advantame). Based on the
data from studies with simulated gastric and intestinal fluids, the majority
of this conversion occurs rapidly in the intestinal tract prior to absorption.
Maximum plasma concentrations of advantame-associated radioactivity
after oral dosing in rodents are reached within 15–45 minutes (Tmax); this is
similar to the Tmax in fasted humans (1.25 hours) and contrasts with the Tmax
of 6–8 hours in fasted dogs. Overall bioavailability following oral doses is
estimated to be less than 10% in rats, 8–15% in dogs and approximately 6%
in humans.
Following oral dosing with radiolabelled advantame, the majority of advan-
tame and/or its metabolites are found to be associated with the gastrointestinal
tract, in particular the stomach and small and large intestine. The low levels
of advantame-associated radioactivity found outside the gastrointestinal tract
were consistent with the presence of these materials in plasma and provided
no indication of tissue-specific distribution or accumulation. Based on auto-
radiography studies, advantame and/or its metabolites were not detected in
the placentas or fetuses of pregnant rats after oral dosing. Human volunteers
ingesting advantame daily over a 12-week period demonstrated no evidence
of accumulation.
12
Advantame acid (de-esterified advantame) is generally the predominant
metabolite found in plasma, urine and faeces of all species except dogs,
where advantame acid accounts for a low proportion (<1%) of the total
absorbed dose in plasma. In this species, a sulfate conjugate was postulated to
be the predominant metabolite in plasma based on high-performance liquid
chromatographic and mass spectrometric analyses. This conjugate, together
with possible enterohepatic circulation of other metabolites, likely explains
the longer terminal half-life of plasma radioactivity in the dog (advantame-
associated radioactivity half-life of 74–85 hours in dogs compared with
6.0–8.1 hours in rats and 3.9 hours in humans). Sulfate conjugates were not
detected in the plasma of rats or humans.
L-Phenylalanine methyl ester was identified as a minor (<1.0%) degradation
product of advantame in beverages under low-pH storage conditions. When
ingested, L-phenylalanine methyl ester would hydrolyse to L-phenylalanine
and methanol. However, it was considered that any potential exposure to either
chemical would be insignificant compared with the usual dietary exposure to
these chemicals.
The oral median lethal dose (LD50) of advantame administered by gavage
to rats was greater than 5000 mg/kg body weight (bw). The subchronic and
chronic toxicity of dietary advantame was assessed in mice (13 and 104
weeks), rats (4, 13 and 104 weeks) and dogs (4, 13 and 52 weeks). No-
observed-adverse-effect levels (NOAELs) for each of these studies were
established on the basis of an absence of adverse effects at the highest con-
centration tested (50 000 mg/kg in the diet, equal to doses ranging from
approximately 2000 to 7400 mg/kg bw per day). A common observation in
the subchronic and chronic rodent feeding studies was a significant decrease
in body weight gain associated with the high-dose group (5% advantame
in the diet) with an associated reduction in feed conversion efficiency, but
not feed intake. A significant reduction in final body weight gain compared
with controls, however, was noted in female mice in the high-dose group
(50 000 mg/kg diet) in the chronic bioassay and occurred in the absence of
a decrease in feed conversion efficiency. As this effect was mainly related to
a decrease in body weight gain observed in senile mice (>78 weeks of age)
and there was no significant difference in the final mean body weights, in the
absence of any other adverse effects, the reduced body weight gain observed
at the highest dose was attributed to the relatively high concentration of a
non-caloric substance in the diet.
In the chronic bioassays designed to assess potential carcinogenicity in mice
and rats, there was no treatment-related increase in tumour incidence at
advantame doses of up to 5693 and 2621 mg/kg bw per day, respectively.
There was no evidence of genotoxicity in any of the in vitro or in vivo tests
13
conducted with advantame. A number of minor degradation products of
advantame (formed at levels of less than 1%) were also tested for genotoxic-
ity potential in vitro. Only advantame-imide gave a weak positive response
in vitro, which was not confirmed on in vivo testing.
In a two-generation study of reproductive toxicity in rats, there were no treat-
ment-related effects on reproductive parameters (mating performance, fertility,
gestation length/index and sperm quality), litter observations (size, survival,
sex ratio and pup body weight) or measures of postnatal offspring development
at dietary concentrations of advantame up to 50 000 mg/kg (equal to advantame
doses in the range of 4000–6000 mg/kg bw per day during the period prior to
mating and gestation and over 8000 mg/kg bw per day during lactation). The
developmental toxicity of advantame was examined in rats (diet) and rabbits
(gavage) at advantame doses of 0, 465, 1418 or 4828 mg/kg bw per day and
0, 500, 1000 or 2000 mg/kg bw per day, respectively. In neither species was
there any evidence of embryotoxicity or teratogenicity up to the highest dose
tested. In rats, there was a transitory decrease in feed consumption at the start
of treatment, resulting in lower body weight gain and subsequent lower final
body weights in high-dose dams; however, there was no adverse effect on sur-
vival, growth or fetal development. In the main rabbit developmental toxicity
study, clinical signs of toxicity (lethargy, loss of coordination and locomotion,
inappetence and body weight loss) leading to humane sacrifice were observed
in the 1000 mg/kg bw per day (n = 1) and 2000 mg/kg bw per day (n = 5) dose
groups. Necropsy findings of these animals included incidences of distended
caecum with or without haemorrhagic walls, congestion of the gastrointesti-
nal tract, kidneys reported with punctate cysts and/or foci on the surfaces and
bladder filled with green-coloured urine. Based on the available data, it could
not be concluded that the clinical symptoms necessitating humane sacrifice in
maternal animals in the mid- and high-dose groups were not treatment related.
In the high-dose group, there was an observation of an approximate 2-fold
increase in post-implantation loss compared with controls. However, the Com-
mittee noted that this effect occurred without a significant reduction in the
number of live offspring per litter and was within the historical control range
for this strain of rabbits. Based on the similarity in clinical signs observed in
gravid rabbits subject to humane sacrifice in the 1000 and 2000 mg/kg bw per
day dose groups, a NOAEL for maternal toxicity of 500 mg/kg bw per day was
assigned, whereas the NOAEL for developmental effects was 2000 mg/kg bw
per day, the highest dose tested.
Studies of human tolerance of advantame included a single-dose pharma-
cokinetic study, a 4-week study in healthy males and a 12-week study in male
and female diabetic subjects. There were no treatment-related adverse effects
or withdrawals during the study periods. Advantame did not affect plasma
levels of glucose or insulin in healthy subjects, exacerbate glucose tolerance
14
or insulin resistance or affect levels of glycosylated haemoglobin in diabetic
subjects with acceptable blood glucose control. The consumption of a single
or repeated dose of advantame up to 0.5 mg/kg bw per day was considered to
be well tolerated by both healthy and diabetic individuals.
Assessment of dietary exposure

Advantame is intended for use as a high-intensity sweetener in a number
of food categories, including use in tabletop sweeteners, where there are
existing provisions in regulations for aspartame. At low concentrations,
advantame can also be used as a flavour enhancer. Application of a modified
budget method as a screening method indicated a theoretical maximum use
level of 400 mg/kg for advantame, assuming use in one quarter of the food
supply and half the beverage supply and an acceptable daily intake (ADI)
for advantame of 0–5 mg/kg bw, as established by Food Standards Australia
New Zealand (FSANZ) in 2011. The proposed use levels for advantame in
chewing gum and tabletop sweeteners were at or above this maximum level,
so further investigations were made.
In an additional screening method, mean dietary exposures to advantame
were predicted, assuming total sugar replacement by advantame in the food
supply and a sweetness relative to sucrose of 20 000:1. Using per capita data
for the food group “Sugar, honey and candies” for the 17 GEMS/Food Clus-
ter Diets (see Annex 3 for the countries included in the 17 Cluster Diets) or
reported total sugar intakes for the population of the USA, known to have
one of the highest sugar intakes in the world, the predicted mean population
dietary exposures to advantame were all less than or equal to 0.2 mg/kg bw
per day, assuming a 60 kg body weight. Dietary exposures to advantame
were also predicted assuming replacement of permitted high-intensity sweet-
eners, by calculating the sucrose equivalent intakes derived from their known
use and converting back to advantame use. Predicted mean dietary exposures
to advantame were, as expected, lower than those for total sugar replacement,
at 0.01–0.03 mg/kg bw per day for the general population and 0.03–0.05 mg/
kg bw per day for high consumers, including people with diabetes, assuming
a 60 kg body weight.
Dietary exposures to advantame were predicted from individual food con-
sumption records from national nutrition surveys for a number of popula-
tions with a known use of foods containing high-intensity sweeteners (the
USA, 22 European countries, Australia and New Zealand) and sponsor-
proposed maximum use levels for each jurisdiction, assuming that brand-
loyal consumers always select foods proposed to contain advantame. The
proposed use levels were derived from existing maximum permitted levels
for aspartame in the relevant jurisdiction and adjusting for the relative
15
sweetness of advantame to aspartame of 100:1, with further alterations
based on taste perception, in some cases. The Committee noted that the
maximum use levels proposed by the sponsor for the European Union (EU)
were double those proposed for the USA in some food categories, as an
additional factor of 2 had been applied to the advantame levels derived from
aspartame maximum permitted levels. Sponsor-proposed use levels for the
GSFA were derived by taking the highest value from the regulations in the
USA or EU. Predicted mean dietary exposures to advantame for consumers
only across the different population groups evaluated ranged from 0.03 mg/
kg bw per day (mean consumers aged 15 years and over in New Zealand,
assuming restricted use in beverages and tabletop sweeteners) to 1.45 mg/
kg bw per day (mean consumers aged 2–6 years in Australia, assuming
sponsor-proposed GSFA use levels). Predicted dietary exposures for high
consumers ranged from 0.06 mg/kg bw per day (90th percentile consumers
aged 15 years and over in New Zealand, assuming restricted use in bever-
ages and tabletop sweeteners) to 2.16 mg/kg bw per day (90th percentile
consumers aged 2–6 years in Australia, assuming proposed GSFA use lev-
els). The predicted dietary exposures for European populations and the esti-
mate for Australian children based on applicant-proposed GSFA use levels
tended to be higher than those for the population in the USA due to higher
maximum levels of advantame proposed for similar food categories and
the fact that broader food categories were used in these estimates, which
included foods for which no use of advantame is intended.
Evaluation
On the basis of the available studies, the Committee considered advantame
to be a substance of low oral toxicity across a range of species, including
humans. Appropriately conducted studies indicated that advantame is not
carcinogenic, mutagenic or teratogenic or associated with any reproductive
or developmental toxicity. The main treatment-related effect that was consid-
ered adverse was the occurrence of morbidity that necessitated early humane
sacrifice of dams in the main rabbit developmental toxicity study (where dos-
ing was by gavage) at and above an advantame dose of 1000 mg/kg bw per
day. While these clinical observations were not observed in any other species
dosed with similar levels of advantame via the diet, in the absence of phar-
macokinetic data for gravid rabbits, it could not be concluded that the effect
was not toxicologically relevant. The rabbit is considered the most sensitive
species in the database, with a NOAEL for maternal toxicity of 500 mg/kg
bw per day.
Although a developmental toxicity study is not considered to be representa-
tive of a long-term toxicity study, an additional safety factor was not consid-
ered necessary, based on the lack of adverse effects observed in long-term
16
dietary studies conducted with comparable levels of advantame in different
species.
An ADI of 0–5 mg/kg bw is established for advantame on the basis of a
NOAEL of 500 mg/kg bw per day for maternal toxicity in a developmental
toxicity study in rabbits and use of a 100-fold safety factor for interspecies
and intraspecies variability.
The Committee agreed that the ADI also applies to those individuals with
phenylketonuria, as the formation of phenylalanine from the normal use of
advantame would not be significant in relation to this condition.
Advantame is intended for use as a tabletop sweetener and in a large variety
of solid and liquid foods. Conservative calculations based on its sweetness
potency (20 000 times that of sucrose) suggest that a mean population die-
tary exposure to advantame of less than 0.2 mg/kg bw per day would result
from total sugar replacement in the diet, even for the maximum reported
mean intake of 160 g total sugars per day for the population of the USA,
assuming a 60 kg body weight. Therefore, a total replacement of sugar with
advantame would not lead to the ADI being exceeded. Using national dietary
exposure estimates and making the “worst case” assumption that brand-loyal
consumers always select foods intended to contain advantame at the sponsor-
proposed maximum use levels for broad food categories suggest that the
maximum mean dietary exposure to advantame would be 1.45 mg/kg bw per
day (29% of the upper bound of the ADI), and the maximum high-percentile
dietary exposure would be 2.16 mg/kg bw per day (43% of the upper bound
of the ADI). The Committee considered these predicted dietary exposures to
advantame to be overestimated due to the conservative assumptions made.
The proposed maximum use levels that the Committee considered for advan-
tame for possible inclusion in the GSFA were not expected to lead to dietary
exposures exceeding the upper bound of the ADI for any population group.
A toxicological monograph and a Chemical and Technical Assessment were
prepared.
New tentative specifications were prepared, requesting, by the end of 2015,
information on:
• the suitability of the headspace gas chromatographic method (using
appropriate dissolution solvent) for determination of residual solvents,
published in Volume 4 of the Combined Compendium of Food Additive
Specifications (Annex 1, reference 180), and data, in a minimum of five
batches, using the method;
• an alternative or improved high-performance liquid chromatographic method
for the assay of advantame and advantame acid using a standard curve;
17
• additional data and analytical methods for the determination of palladium
and platinum;
• information on the purity and availability of the commercial reference
standards used in the assay of advantame and advantame acid.
3.1.2 Glucoamylase from Trichoderma reesei expressed in Trichoderma

reesei
Explanation
At the request of CCFA at its Forty-fourth Session (10), the Commit-
tee evaluated the safety of the glucoamylase enzyme preparation (glucan
1,4-α-glucosidase; Enzyme Commission No. 3.2.1.3) from Trichoderma
reesei expressed in T. reesei, which it had not evaluated previously. Glu-
coamylase catalyses the hydrolysis of terminal (1→4)-linked α-D-glucose
residues successively from the non-reducing end of the chain with concomi-
tant release of β-D-glucose in polysaccharide substrates. In this report, the
expression “glucoamylase” refers to the glucoamylase enzyme and its amino
acid sequence, and the expression “glucoamylase enzyme preparation” refers
to the preparation formulated for commercial use. The glucoamylase enzyme
preparation is used as a processing aid in the manufacture of sweeteners such
as high-fructose corn syrup, in baking, in brewing and in the production of
potable alcohol (spirits).
Genetic modification
Glucoamylase is produced from a genetically modified strain of Trichoderma
reesei containing the glucoamylase gene from T. reesei. Trichoderma reesei
is a mesophilic filamentous fungus that is ubiquitous in nature. It has a long
history of use in the production of enzymes used in food processing, includ-
ing enzymes from genetically engineered strains of the organism. Prior to the
introduction of the glucoamylase gene, the T. reesei host strain was geneti-
cally modified through deletion of genes encoding cellobiohydrolase 1 and 2
and endoglucanase 1 and 2, resulting in a strain with a compromised ability
to use cellulose as a carbon source. The modified host strain was then trans-
formed using two expression cassettes containing the glucoamylase gene
from T. reesei. The final recombinant production strain is genetically stable
and free of any antibiotic resistance genes or vector deoxyribonucleic acid
(DNA) used during transformation.

Glucoamylase is produced by submerged aerobic, straight-batch or fed-batch
pure culture fermentation of a genetically modified strain of T. reesei contain-
ing the gene coding for glucoamylase from T. reesei. The enzyme is secreted
18
into the fermentation broth and is subsequently purified and concentrated.
The enzyme concentrate is formulated with glucose, sodium benzoate and
potassium sorbate to achieve the desired activity and stability. The glucoam-
ylase enzyme preparation contains commonly used food-grade materials
and conforms to the General Specifications and Considerations for Enzyme
Preparations Used in Food Processing (Annex 1, reference 154). Glucoamyl-
ase activity is measured in glucoamylase units (GAU). One GAU is defined
as the amount of enzyme that releases 1 g (5.6 mmol) of glucose per hour
from soluble starch substrate (p-nitrophenyl-α-D-glucopyranoside) at pH 4.3
and a temperature of 30 °C. The mean activity of glucoamylase enzyme from
three formulated batches of the enzyme preparation was 523 GAU per gram
of glucoamylase enzyme preparation.
A typical commercial formulation of the glucoamylase enzyme prepara-
tion will contain 10–15% of enzyme as total organic solids (TOS). TOS
includes the enzyme of interest and residues of organic materials, such as
proteins, peptides and carbohydrates, derived from the production organism
during the manufacturing process. The glucoamylase enzyme preparation is
used at concentrations ranging from 0.05 to 0.32 g TOS per kilogram of
food, depending on the proposed application. The glucoamylase enzyme is
expected to be inactivated during processing and hence is not expected to
remain in the final food.
Assessment of potential allergenicity

The glucoamylase from T. reesei was evaluated for potential allergenicity
according to the bioinformatics criteria recommended by FAO/WHO. The
amino acid sequence of glucoamylase was compared with the amino acid
sequences of known allergens. A similarity search in the Structural Database
of Allergenic Proteins (SDAP)1 for matches showing greater than 35% iden-
tity over a window of 80 amino acids produced multiple matches with Sch c
1 protein of Schizophyllum commune. A similar search using the Allermatch
database2 also produced multiple matches with Sch c 1 protein and in addi-
tion produced one match with Pen ch 13 protein in Penicillium chrysogenum.
The Sch c 1 and Pen ch 13 proteins are not identified as food allergens in the
WHO–International Union of Immunological Societies (WHO-IUIS) list of
allergens3. Using SDAP, a search for six contiguous amino acid sequences of
the glucoamylase that could be present in allergenic proteins produced multi-
ple matches as well. The vast majority of these matches are with various sec-
tions of the Sch c 1 protein of Schizophyllum commune, which was expected.
The Sch c 1 protein is not identified as a food allergen in the WHO-IUIS list
1
http://fermi.utmb.edu/SDAP/sdap_who.html
2
http://www.allermatch.org/
3
http://www.allergen.org/
19
of allergens. However, in addition to the Sch c 1 protein, a number of other
allergenic proteins in SDAP were also found to share six contiguous amino
acid sequence identity with glucoamylase. Therefore, the biological relevance
of these matches was examined by investigating the distribution of these six
contiguous amino acid sequences in proteins in general—that is, whether these
sequence segments are present only in allergenic proteins or are present in both
allergenic and non-allergenic proteins. This extended search was performed
using the National Center for Biotechnology Information protein database4,
which contains the sequences of all known proteins. The search demonstrated
that each of these six contiguous amino acid sequences of the glucoamylase
that was found in other allergenic proteins is widely distributed in various
prokaryotic and eukaryotic proteins, including proteins from non‑allergenic
sources, and even edible sources. This indicates that the six contiguous amino
acid sequence matches found between the glucoamylase and various allergenic
proteins occurred by chance and are not likely to be part of any allergenicity-
associated epitopes. Therefore, the Committee considered that oral intake of
glucoamylase is not anticipated to pose a risk of allergenicity.
Toxicological data
In a 13-week study of general toxicity in rats, no treatment-related adverse
effects were seen when glucoamylase enzyme preparation was administered
daily by gavage at doses up to 166 mg TOS per kilogram body weight per
day. The glucoamylase enzyme preparation was not mutagenic in a bacte-
rial reverse mutation assay in vitro and was not clastogenic in an assay for
chromosomal aberrations in human lymphocytes in vitro.

A theoretical “worst case” dietary exposure estimate was made assuming
that the enzyme remains in the food at its maximum concentration following
its use as a processing aid in the production of sugars, bread/bakery items,
beer and spirits and that it is present in 100% of each range of products.
Based on these very conservative assumptions, the estimate of total dietary
exposure was 1.73 mg TOS per kilogram body weight per day. However, the
enzyme is not expected to remain in sugar, beer or spirit products following
purification processes used in their manufacture, and it will be inactivated in
bread and bakery products.
Evaluation
Based on its low toxicity and because it is reasonably anticipated that die-
tary exposure would be very low, the Committee established an ADI “not
4
http://www.ncbi.nlm.nih.gov/protein
20
specified” for the glucoamylase enzyme preparation from T. reesei expressed
in T. reesei used in the applications specified and in accordance with good
manufacturing practice.
A toxicological monograph was prepared.
New specifications and a Chemical and Technical Assessment were prepared.
3.1.3 Glycerol ester of gum rosin
Explanation
At its seventy-first meeting (Annex 1, reference 196), the Committee evalu-
ated glycerol ester of gum rosin (GEGR) for use as an emulsifier/density
adjustment agent for flavouring agents in non-alcoholic beverages and cloudy
spirit drinks. The Committee established a group ADI of 0–25 mg/kg bw
for GEGR and glycerol ester of wood rosin (GEWR). GEGR was evaluated
based on the toxicity data for GEWR, the absence of toxicological effects of
their corresponding non-esterified rosins and the qualitative similarity of the
chemical components of GEGR and GEWR. However, in view of the limited
toxicity data available for GEGR and the submission of only the summarized
results of two 90-day oral toxicity studies in rats, the Committee concluded
that the full reports of the two 90-day oral toxicity studies with GEGR were
needed to confirm the validity of the comparison of GEGR with GEWR.
Further, at the seventy-first meeting, the specifications for GEGR were made
tentative pending submission of additional data regarding the identity and
compositional analysis of GEGR to establish the extent of the chemical
similarity between GEGR and GEWR.
At its seventy-fourth meeting (Annex 1, reference 205), the Committee
noted that the requested full reports of the two 90-day oral toxicity studies
on GEGR in rats had not been provided and that the validity of evaluating
GEGR on the basis of toxicological data on GEWR still required confirma-
tion. The Committee withdrew the group ADI for GEGR and GEWR and
established a temporary group ADI for GEGR and GEWR of 0–12.5 mg/
kg bw. The Committee noted that the temporary group ADI would be with-
drawn if compositional information on GEGR as well as the full reports of
the two 90-day oral toxicity studies on GEGR in rats were not submitted by
the end of 2012.

GEGR is a complex mixture of glycerol diesters and triesters of resin acids
from gum rosin, with a residual fraction of glycerol monoesters. Gum rosin
is an exudate of living pine trees.
21
Although the submitted analytical data included summarized information
in relation to the composition of free resin acids and neutrals (non-acidic
saponifiable and unsaponifiable substances) in GEGR, the Committee noted
that the information on the composition and ester distribution of GEGR
was incomplete and therefore could not confirm the claimed similarities to
GEWR.
Evaluation
For the present meeting, the requested two unpublished 90-day oral toxicity
studies on GEGR in rats were not submitted. Furthermore, complete infor-
mation on the composition of GEGR was not submitted. As the requested
data were not submitted, the Committee withdrew the temporary group ADI
of 0–12.5 mg/kg bw for GEGR and GEWR.
The specifications were maintained as tentative pending the submission of
additional information by the end of 2014. Additional data are requested to
characterize GEGR in commerce in relation to the composition of 1) the
refined gum rosin currently used as the source rosin with regard to the
levels (%) of resin acids and neutrals, 2) the glycerol ester of gum rosin
with regard to the levels (%) of a) glycerol esters, b) free resin acids and
c) neutrals and 3) the total glycerol esters of resin acids with regard to the
levels (%) of a) glycerol monoesters and b) the sum of glycerol diesters and
triesters (assay). Validated methods for the determination of the substances
considered in the specifications are also required.
No toxicological monograph was prepared.
3.1.4 Glycerol ester of tall oil rosin
Explanation
Glycerol ester of tall oil rosin (GETOR) was evaluated by the Committee
at its seventy-first meeting (Annex 1, reference 196) for proposed use as an
emulsifier/density adjustment agent for flavouring agents in non-alcoholic
beverages. The Committee concluded that, in principle, the data for GEWR,
which had been evaluated previously, could be used for the evaluation of
GETOR, provided the respective compositional data were sufficiently similar.
Because the information on the composition of GETOR was inadequate to
conclude that GETOR is sufficiently similar to GEWR, the Committee could
not complete the evaluation and requested additional compositional data on
the product in commerce, in order to clarify the extent and significance of
any differences relative to other glycerol esters of rosins.
At the seventy-fourth meeting of the Committee (Annex 1, reference 205),
limited data on the composition of GETOR were provided. The claimed
22
s
imilarities between GETOR and GEWR could not be confirmed. The
Committee concluded that additional data were required to characterize the
GETOR in commerce in relation to the composition of 1) the refined tall oil
rosin used as the source rosin, 2) the glycerol esters of tall oil rosin, 3) the
total glycerol esters of resin acids and 4) the neutrals. Validated methods for
the determination of the substances considered in the specifications were
also requested.
Evaluation
For the present meeting, no data on GETOR were submitted, and the Sec-
retariat was informed that this compound is no longer supported by the
previous data sponsor. Therefore, no evaluation was performed.
The tentative specifications were withdrawn.
3.1.5 Glycerol ester of wood rosin
Explanation
GEWR was previously considered by the Committee at its eighteenth, twen-
tieth, thirty-third, thirty-seventh, forty-fourth, forty-sixth, seventy-first and
seventy-fourth meetings (Annex 1, references 35, 41, 83, 94, 116, 122, 196
and 205). At its forty-sixth meeting, the Committee established an ADI of
0–25 mg/kg bw for GEWR. At its seventy-first meeting, the Committee
extended this ADI to a group ADI of 0–25 mg/kg bw for GEGR and GEWR.
Prior to the seventy-fourth meeting, GEWR was evaluated based on the
assumption that the substance was obtained from just one Pinus species (Pinus
palustris). However, information received by the Committee at its seventy-
fourth meeting indicated that the product in commerce is also produced from
the species Pinus elliottii. Considering that the rosin source may have an impact
on the composition of the final glycerol ester, the Committee decided that the
specifications should include all the Pinus species from which the wood rosin is
obtained. As the submitted information on GEWR did not adequately charac-
terize the product in commerce, the Committee decided to withdraw the group
ADI of 0–25 mg/kg bw for GEGR and GEWR and established a temporary
group ADI of 0–12.5 mg/kg bw by applying an additional uncertainty factor
of 2. The Committee requested additional data to characterize the GEWR in
commerce in relation to the composition of 1) the refined wood rosin currently
used as the source rosin for the production of GEWR, 2) the glycerol esters
of wood rosin, 3) the total glycerol esters of resin acids and 4) the neutrals.
Validated methods for the determination of the substances considered in the
specifications were also requested. The Committee noted at the seventy-fourth
23
meeting that the temporary group ADI would be withdrawn unless additional
compositional information on GEWR from Pinus elliottii and Pinus palustris
as well as the full reports of two 90-day oral toxicity studies on GEGR in rats
were submitted by the end of 2012.

GEWR is a complex mixture of glycerol diesters and triesters of resin acids
from wood rosin, with a residual fraction of glycerol monoesters. Wood rosin
is obtained by solvent extraction of aged pine stumps.
The sponsor submitted the requested information on the composition of the
wood rosin and GEWR, based on analyses of five production samples of
the wood rosin and GEWR. The wood rosin was composed of resin acids
and neutrals. GEWR was composed mainly of glycerol diesters and triesters
(78.3–83.9%) and neutrals (11.4–17.6%), with lesser amounts of glycerol
monoesters (1.5–3.2%) and free resin acids (2.3–2.8%). Further, the spon-
sor informed the Committee that throughout the history of the manufacture
of GEWR, the wood rosin feedstock has always been based on a mixture
of P. palustris and P. elliottii, which occur naturally in overlapping regions
of the south-eastern USA. Considering this statement, in addition to the
submitted compositional information, the Committee recognized that the
GEWR used for each of the toxicological studies, just as the product now in
commerce, was based on a mixture of the two species.
Based on this recognition, the specifications were revised to exclude both
assays and limits for glycerol monoesters and neutrals and further to include
gas chromatographic analysis for the confirmation of the presence of glyc-
erol and major resin alcohols, abietic alcohols and dehydroabietic alcohols,
generated by reductive reaction of GEWR.
Evaluation
At the present meeting, new information, including compositional data on
GEWR, was submitted. Based on this information, the Committee concluded
that the current product in commerce is equivalent to the GEWR with which
the toxicological studies have been performed.
For the present meeting, the requested data on GEGR were not submitted.
The Committee therefore withdrew the temporary group ADI of 0–12.5 mg/
kg bw for GEGR and GEWR and re-established the ADI of 0–25 mg/kg bw
for GEWR.
The existing tentative specifications were revised, and the tentative status
was removed. A Chemical and Technical Assessment was prepared.
24
3.1.6 Nisin
Explanation
Nisin is a mixture of antimicrobial polypeptides produced by Lactococcus
lactis subsp. lactis. Nisin preparations are currently used in commerce as an
antimicrobial preservative in processed cheeses, pasteurized dairy products
and processed meats. The Committee evaluated the safety of nisin at the
twelfth meeting of JECFA (Annex 1, reference 17) and established an ADI
of 0–33 000 units of nisin per kilogram body weight.
At its Forty-fourth Session (10), CCFA requested JECFA to verify the calcula-
tion of the ADI for nisin; clarify the basis of the ADI for nisin; and provide the
calculation to convert units of nisin to milligrams of nisin.
In the original JECFA evaluation of nisin, the ADI of 0–33 000 units of nisin
per kilogram body weight was derived from the highest dose in a 2-year
repeated-dose toxicity study (11), which the Committee interpreted as being
3 330 000 units of nisin per kilogram of body weight, but which was actu-
ally 3 330 000 units of nisin per kilogram of feed. As the ADI in the original
JECFA evaluation was derived incorrectly, the present Committee undertook
a re-evaluation of nisin, taking previously evaluated as well as new studies
into account.

Nisin is a mixture of closely related antimicrobial polypeptides that are 34
amino acids in length. Nisin is produced by strains of Lactococcus lactis
subsp. lactis under appropriate fermentation conditions. The major poly-
peptide from the fermentation is nisin A. Nisin is produced in a sterilized
medium of non-fat milk solids or non-milk-based fermentation sources, such
as yeast extract and carbohydrate solids. The fermentation process is con-
trolled for time and pH, until optimum nisin production has been achieved.
Nisin is recovered, concentrated and purified from the fermentation medium
by various methods, such as sterile injection, membrane filtration, acidifica-
tion, salting out or spray-drying. The purified nisin is then standardized to
make nisin preparation at the desired activity level. The remaining compo-
nents of the preparation are milk solids and products of fermentation, which
include proteins and carbohydrates. Nisin is commercially available as nisin
preparation and contains commonly used food-grade materials. A typical
batch of a commercial nisin preparation contains 2.5% (weight per weight
[w/w]) nisin and approximately 75% (w/w) sodium chloride.
The activity of nisin in commercial nisin preparation has been described in
the literature in Reading Units (RU) and International Units (IU). In 1970,
the WHO Committee on Biological Standardization established the IU as
25
the international reference for units of nisin activity. One IU was defined
as the amount of nisin required to inhibit the growth of one bacterial cell in
1 ml of broth. This amount is 0.025 µg of nisin (= 1 IU). Therefore, 1 µg of
nisin is equivalent to 40 IU. The Committee at the current meeting reviewed
the scientific literature in order to harmonize the RU and IU and concluded
that they are equivalent (i.e. 1 IU is the same as 1 RU). The Committee at
the current meeting clarified that the specification for the activity of nisin is
reported as International Units and provided a conversion factor to obtain
the quantity of active nisin in a given sample. The revised JECFA specifica-
tions for nisin include this conversion. The revised assay for nisin activity in
the JECFA specifications is not less than 900 IU of nisin per milligram or
not less than 22.5 µg of nisin per milligram. The revised specifications also
removed the synonym “nisin preparation”.
Toxicological data
In vitro studies demonstrated that nisin is inactivated by α-chymotrypsin.
Nisin administered by gavage to rats was also hydrolysed and inactivated
in the intestinal tract, with no biologically active nisin being detected in the
colon or caecum. In a gavage study in dogs dosed with nisin (test material
description not available) at 1000 mg/kg bw twice per day for 28 days, bio-
logically active nisin was detected in blood serum in one of six dogs at a level
of 0.54 µg/ml (limit of detection: 0.45 µg/ml). As the experimental design
did not involve multiple time points to confirm the presence of biologically
active nisin in plasma, the Committee did not find the results of this gavage
dosing study to be conclusive evidence that nisin in food would result in
systemic exposure to biologically active nisin.
A range of studies on the acute toxicity, short- and long-term toxicity, geno-
toxicity, and reproductive and developmental toxicity of nisin was taken into
consideration for the safety assessment.
The oral LD50 of nisin preparation in mice was 6950 mg/kg bw, and the oral
LD50 of purified nisin in rats was greater than 2000 mg/kg bw. The toxicity of
orally administered (diet or gavage) nisin or nisin preparation was assessed in
short-term studies in mice, short- and long-term studies in rats and short-term
studies in dogs. A common observation in animals treated with nisin prepara-
tions (with ~75% [w/w] sodium chloride) was a significant increase in the
absolute and relative kidney weights, coupled with high water consumption
and increased urination. The increased kidney weight and minimal squamous
cell hyperplasia of the limiting ridge in the forestomach that were seen in these
studies in the nisin preparation–treated groups were also seen in the sodium
chloride control groups, and these effects are known to be typical of high
sodium chloride treatment. Slight but statistically significant changes in hae-
matological parameters at high doses of nisin (2000 mg of purified nisin per
26
kilogram body weight per day in two studies and a 5% dietary level of nisin
preparation in another study) were inconsistent in direction and magnitude.
In a 90-day toxicity study, rats were fed nisin preparation (with a nisin A
potency of 3000 IU/mg, corresponding to a 7.5% nisin A content) in the diet at
0%, 0.2%, 1.0% or 5.0% (equal to 0, 117, 586 and 2996 mg of nisin prepara-
tion per kilogram body weight per day for males and 0, 129, 638 and 3187 mg/
kg bw per day for females, respectively). A sodium chloride reference group
was given sodium chloride at a dietary level of 3.712%, which was the same as
that in the 5.0% nisin A diet. Significant increases in the absolute and relative
kidney weights, coupled with high water consumption, increased urination and
minimal squamous cell hyperplasia of the limiting ridge in the forestomach,
were observed. These changes were also noted in the sodium chloride refer-
ence group and were considered to be related to sodium chloride intake. Other
parameters that showed a statistically significant increase in the high-dose group
were the red blood cell parameters (red blood cell count, haemoglobin, mean
corpuscular haemoglobin, mean corpuscular volume, haematocrit value). The
increase in these parameters was less than 4% and within the range of historical
data and was not considered to be toxicologically relevant. Based on the lack
of treatment-related adverse effects in the highest dose group, the NOAEL was
identified as 5.0% of the nisin preparation in the diet, which was equal to 2996
mg/kg bw per day of the nisin preparation, or 224.7 mg/kg bw per day of nisin.
In three one-generation reproductive toxicity studies, the reproductive per-
formance of the nisin-treated rats was unaffected. There were no perinatal
or postnatal effects. Growth of the pups was normal and similar to that of
the control group. In a three-generation reproductive toxicity study, a nisin
preparation containing 2.5% nisin A was fed to rats in a standard diet con-
taining 0%, 0.2%, 1.0% or 5.0% nisin preparation for 26 weeks. A further
group of animals received a diet containing 3.8% sodium chloride, which
was equivalent to the sodium chloride content of the 5.0% diet group. No
treatment-related changes were observed in reproductive performance as
assessed by pregnancy rate, gestation length, postpartum litter loss, litter size,
mortality or necropsy findings. There were no treatment-related changes in
organ weights or histopathology at the end of the study. A decrease in body
weight gain was observed in males in the F0 and F2 generations, but not in
the F1 generation. Decreased body weight gain was not observed in females.
Furthermore, a decrease in body weight gain in rats of both sexes was not
observed in other studies. Therefore, the Committee considered this finding
to be unrelated to nisin treatment.
In a developmental toxicity study in rats administered purified nisin by gav-
age at doses up to 50 mg/kg bw per day, no effects on any developmental
end-points were observed.
27
Purified nisin was not genotoxic in reverse mutation, chromosomal aberra-
tion, mouse lymphoma or mouse bone marrow micronucleus assays. Nisin
was not carcinogenic in the 2-year rat study at dietary concentrations up to
3.33 million units of nisin per kilogram diet. Based on the authors’ statement
that 1 g of nisin had an activity of approximately 40 million units and their
assumptions that the average weight of the rats was 250 g and the average
feed consumption of the rats was 15 g, this is equivalent to 83.3 mg of nisin
per kilogram diet, or 5.0 mg/kg bw per day. This was the study from which
the Committee derived the ADI at the twelfth meeting.

Nisin has been used commercially for over 25 years in a number of food
types, primarily processed cheese and meat products. The Committee
received information concerning dietary exposure patterns from one spon-
sor and independently obtained additional published information. The spon-
sor data had been submitted as part of a premarket evaluation of nisin for
expanded uses in Japan in the mid-2000s. The additional information was
taken from regulatory publications in Australia/New Zealand, Europe and
the USA. Four national/regional estimates of dietary exposure to nisin were
reviewed by the Committee: from the EU, Australia/New Zealand, Japan and
the USA. The estimate of mean consumers-only dietary exposure for the EU
from consumption of cheese, cream, desserts and egg products was 0.008 mg/
kg bw per day, with exposure at the 97.5th percentile of 0.026 mg/kg bw per
day. From FSANZ, the estimated consumers-only mean dietary exposures to
nisin from consumption of cheese, cream, meat products, sauces, toppings
and mayonnaise were 0.009 mg/kg bw per day (all ages) and 0.02 mg/kg
bw per day (2- to 6-year-olds). Estimated consumers-only 95th percentile
dietary exposures to nisin were lowest for New Zealanders aged 15 years and
above, at 0.03 mg/kg bw per day, and highest for Australian children aged
2–6 years, at 0.07 mg/kg bw per day. The consumers-only dietary exposure
estimate from the USA from consumption of cheese spreads, dressings, egg
products and processed meat products was 0.04 mg/kg bw per day at the
mean. The Committee also noted that a letter responding to a Generally Rec-
ognized as Safe notification in the USA for nisin use only in frankfurters
included an estimate of consumer-only mean dietary exposure to nisin of
0.6 mg/day, or 0.01 mg/kg bw per day, with exposure at the 90th percentile of
1.1 mg/day, or 0.02 mg/kg bw per day. The Japanese per capita estimate was
from consumption of cheeses, buns, meat and egg products, tofu and miso
and was reported as 2.06 mg/person per day or approximately 0.04 mg/kg
bw per day for a 50 kg individual. The FSANZ, Japanese and USA estimates
were consistent and higher than the EU estimate due to broader food catego-
ries in which nisin could be applied. The Committee concluded that the use
28
of a dietary exposure of 0.07 mg/kg bw per day (95th percentile, Australian
children, 2–6 years old) was appropriate for the safety evaluation of nisin.
Evaluation
At the meeting, the Committee provided clarifications on the identity of nisin
and on the units of activity of nisin and a calculation to convert from Inter-
national Units of nisin to micrograms of nisin based on the available data on
different forms of nisin in commerce.
On the basis of the available studies, the Committee considered nisin to be
a substance of low oral toxicity. Ingested nisin is inactivated in the upper
part of the intestinal tract. Nisin is not carcinogenic or mutagenic and is not
associated with any reproductive or developmental toxicity. The 2-year tox-
icity study was not considered to be an appropriate basis for establishing an
ADI because it was not conducted to current standards and did not include
an appropriate saline control group and because a number of later studies
investigated higher doses of nisin.
After evaluating the new studies as well as the previously reviewed studies, the
Committee concluded the 13-week subchronic toxicity study to be the pivotal
study in the current safety evaluation of nisin because it was a higher-dose
study and it took into consideration more parameters compared with other
studies, such as the three-generation reproductive toxicity study. Based on the
observation that there were no treatment-related adverse effects at the high-
est concentration tested, the 5% dietary level of nisin preparation (containing
7.5% nisin), a NOAEL of 224.7 mg of nisin per kilogram body weight per day
was identified. Applying a safety factor of 100 to the NOAEL to account for
interspecies and intraspecies variability, the Committee established an ADI for
nisin of 0–2 mg/kg bw. The Committee did not consider it necessary to use an
additional safety factor to account for the short duration of the study because
no compound-related effects were observed at any dose in any of the other
studies, including the reproductive toxicity study, and because ingested nisin is
degraded in the upper part of the intestinal tract, such that systemic exposure
to nisin is not likely to occur.
The highest estimated dietary exposure of 0.07 mg of nisin per kilogram
body weight per day determined at the current meeting did not exceed the
upper bound of the ADI.
The Committee withdrew the previous ADI of 0–33 000 units of nisin per
kilogram body weight established at the twelfth meeting.
A toxicological monograph was prepared.
The specifications for nisin were revised.
29
3.1.7 Octenyl succinic acid modified gum arabic
Explanation
At its seventy-first meeting (Annex 1, reference 196), the Committee evalu-
ated the toxicological and chemical and technical data for octenyl succinic
acid (OSA) modified gum arabic. In view of the similarities between OSA
modified gum arabic and the parent gum arabic, toxicological information
for gum arabic was included in the toxicological monograph. At that meet-
ing, the Committee decided to allocate a temporary ADI “not specified” to
OSA modified gum arabic, pending submission of data by the end of 2011
showing hydrolysis of OSA modified gum arabic in the gastrointestinal
tract to confirm the validity of using toxicological data on gum arabic in the
evaluation of OSA modified gum arabic.
At the seventy-fourth meeting (Annex 1, reference 205), the Committee eval-
uated new data on the hydrolysis of OSA modified gum arabic and reviewed
the specifications. The Committee concluded that the results from the experi-
ments on the hydrolysis of OSA modified gum arabic did not unequivocally
demonstrate that OSA modified gum arabic hydrolyses completely in the
stomach into gum arabic and OSA. Furthermore, the hydrolysis experiments
showed inconsistencies with the reported stability of OSA modified gum
arabic in food. Therefore, the Committee deferred further evaluation of OSA
modified gum arabic and requested that the following data be provided by
the end of 2013:
• data resolving the concern about the stability of OSA modified gum
arabic in food;
• data confirming that OSA modified gum arabic is (completely) hydro-
lysed in the gastrointestinal tract, to confirm the validity of using gum
arabic data in the evaluation of OSA modified gum arabic.
The temporary ADI was retained and the specifications were revised with
changes in the test methods for the degree of esterification and for residual
OSA content.
At the present meeting, new data on the hydrolysis of OSA modified gum
arabic in simulated gastric fluid, as well as the stability of OSA modified
gum arabic in food, were evaluated by the Committee. The specifications
were also reviewed.
Biochemical data
In an in vitro test, the hydrolysis of OSA modified gum arabic (lot no. 19705,
purity not reported) in simulated gastric fluid (0.2% weight per volume [w/v]
sodium chloride in 0.7% volume per volume [v/v] hydrochloric acid, without
30
pepsin, pH 1–2), simulated intestinal fluid (without pancreatin, USP XXII
formulation, pH 7.5) and water was investigated. OSA modified gum arabic
at a concentration of 3 mg/ml was incubated in simulated gastric fluid, simu-
lated intestinal fluid or water for 0 or 60 minutes at 37 °C. The reactions were
terminated by addition of a neutralizing agent, after which the OSA con-
tent was analysed by liquid chromatography with tandem mass spectrometry
(LC-MS/MS).
Incubation of OSA modified gum arabic at 3 mg/ml in simulated gastric fluid,
simulated intestinal fluid or water for 0 and 60 minutes resulted in the forma-
tion of OSA free acid at concentrations of 353 and 364 μmol/l in simulated
gastric fluid; 335 and 321 μmol/l in simulated intestinal fluid; and 275 and
292 μmol/l in water, respectively. In view of the high level of OSA free acid
at 0 minutes, the study author concluded that hydrolysis of OSA modified
gum arabic was immediate, with no apparent time dependence. However,
the Committee questioned the effectiveness of the addition of a neutralizing
agent to the reaction mixture to terminate the reaction, as marked hydrolysis
of OSA modified gum arabic occurred in water. Following “termination” of
the reaction, the mixture is processed for analysis by LC-MS/MS, which is
reported to take at least 10 minutes and possibly longer. The formation of
OSA free acid at a concentration of 353–364 μmol/l is equal to about 0.08
mg/ml, or 2.7% w/w of the starting material, which contains maximally 3%
of OSA, according to the specifications. The study author considered that the
study indicates virtually complete hydrolysis in simulated gastric fluid (12).
Stability in food
Data on the stability of OSA modified gum arabic in two model food systems
were presented (13). The two model food systems used were beverage and salad
dressing emulsions. Results of these studies demonstrate that OSA modified
gum arabic is effective in forming stable emulsions in these foods over a given
shelf life. The emulsion stability in the beverage model system was evaluated
through particle size analysis at three pH levels (2.5, 3.6 and 4.5) and two tem-
peratures (room temperature and 45 °C), ring test (over 30 days) and backscat-
tering analysis (over 21 days). A standard formula for French salad dressing
was used as the model system for the salad dressing. Emulsion stability was
evaluated using backscattering analysis and viscosity. Backscattering analy-
ses for 17.5% and 30% oil French dressings containing OSA modified gum
arabic demonstrate stability over 4.4 and 5 months, respectively. Studies also
demonstrate droplet size stability over 30 days in addition to stable viscosity.
Evaluation
The Committee noted that complete hydrolysis of OSA modified gum ara-
bic under neutral pH conditions in simulated intestinal fluid or water, as
31
reported in the study submitted for the present meeting, was at variance with
the claimed stability of the OSA ester linkage in aqueous solutions at the pH
range of foods and beverages. The Committee considered that the spontane-
ous hydrolysis of OSA modified gum arabic in water was unlikely to occur,
which therefore raised doubts about the validity of the observed hydrolysis in
the presence of gastrointestinal enzymes. In view of this, the Committee con-
sidered that the present study does not unequivocally demonstrate that OSA
modified gum arabic hydrolyses completely in the stomach into gum arabic
and OSA and that the validity of using toxicological data on gum arabic in
the evaluation of OSA modified gum arabic had not been confirmed.
The Committee also considered that the presented data demonstrate that
OSA modified gum arabic provided a stable emulsion in the two model food
systems evaluated. However, the data did not unequivocally demonstrate that
the OSA modified gum arabic, at the molecular level, is stable in food and
beverages.
The Committee noted that ongoing studies on the stability of OSA modified
gum arabic in food may provide further information on its chemical state in
food and aqueous solutions, which could help to explain the contradictory
results of the hydrolysis study.
Therefore, the Committee decided to retain the temporary ADI “not speci-
fied” pending submission of additional data on the stability of OSA modified
gum arabic in food by the end of 2013.
An addendum to the toxicological monograph was not prepared.
The Committee also reviewed the specifications and noted that the purity test
of degree of esterification in the current specifications should be replaced
by the degree of substitution and requested information for an analytical
method to measure the degree of substitution and results of the analysis of
at least five commercially available batches. The specifications were made
tentative pending submission of these data by the end of 2013.
3.2 Revision of specifications
3.2.1 Annatto extracts (solvent-extracted bixin and solvent-extracted

norbixin)
At the present meeting, the Committee considered information on the lev-
els of residual solvents in commercial preparations of solvent-extracted
annatto extracts of bixin and norbixin. The Committee noted that levels of
methanol, isopropanol or acetone in many commercially available prepa-
rations exceeded those in the current specifications by a large margin. In
the absence of sufficient information on residual solvent levels received
32
in response to the call for data, the Committee decided not to revise the
provision for residual solvents in these substances. The Committee rec-
ommended that manufacturers supply residual solvent data from at least
five batches of each of the solvent-extracted bixin and norbixin products
to support the possible revision of the provision for residual solvents. The
existing specifications were maintained.
The Committee also considered the suitability of the general method for the
determination of residual solvents published in Volume 4 of the Combined
Compendium of Food Additive Specifications (Annex 1, reference 180) for
the analysis of solvent-extracted bixin and norbixin products. The Commit-
tee concluded that neither solvent listed in the method is suitable for the
analysis of solvent-extracted bixin and norbixin by headspace gas chroma-
tography. Accordingly, the Committee considered a method to allow the use
of dimethyl formamide as the dilution solvent. This method will be pub-
lished as tentative in FAO JECFA Monographs 14 (2013) and included in the
online version of Volume 4 of the Combined Compendium of Food Additive
Specifications (Annex 1, reference 180). In order to evaluate the suitability
of the method for the determination of residual solvents in annatto extracts
dissolved in dimethyl formamide, the Committee recommends that manu-
facturers provide results from the analysis of samples of solvent-extracted
bixin and norbixin products using both methods.
3.2.2 Benzoe tonkinensis
The specifications of Benzoe tonkinensis prepared at the seventy-fourth meet-

ing of the Committee (Annex 1, reference 205) were made tentative pend-
ing submission of the following data: complete composition of the ethanolic
extract, data on microbiological contaminants, data on inorganic contaminants
(lead, arsenic, antimony, chromium, mercury and cadmium) and an analytical
method to distinguish between Benzoe tonkinensis and Benzoe sumatranus.
In response to a call for data, the Committee at the present meeting received
only some of the requested data, including information on inorganic contam-
inants and a gas chromatographic–mass spectrometric method to determine
cinnamic acid, used to distinguish between Benzoe tonkinensis and Benzoe
sumatranus. The Committee noted that the data provided were not sufficient
to revise the specifications and decided that the tentative specifications will
be withdrawn if the complete data on the composition of the ethanolic extract
and microbiological contaminants are not received by the end of 2013.
3.2.3 Food additives containing aluminium and/or silicon
The Committee at its seventy-sixth meeting (Annex 1, reference 211)

reviewed the analytical methods for food additives containing aluminium
33
and/or silicon and found that some test methods use potentially corrosive or
hazardous reagents that may not be permitted in current laboratory safety
protocols. The Committee also noted that aluminium silicate, calcium sili-
cate and sodium aluminosilicate have no provisions for assay. Calcium alu-
minium silicate, aluminium silicate, calcium silicate and silicon dioxide
were placed on the agenda of the current meeting for revision of their speci-
fications. In addition, the Committee, at its current meeting, agreed that it
was appropriate to revise the specification for sodium aluminosilicate, while
noting that it had not been included in the call for data.
The Committee received limited information on the assay of aluminium sili-
cate, calcium silicate and sodium aluminosilicate. However, other requested
information was not received.
Specifications were revised and made tentative pending the submission of the
requested information. Information required includes composition; methods
of manufacture; data on loss on drying and loss on ignition; impurities (lead,
cadmium, arsenic and mercury) soluble in hydrochloric acid (0.5 mol/l);
and suitability of the proposed inductively coupled plasma – atomic emis-
sion spectrophotometric (ICP-AES) method for assay, as well as data on the
assay. Details on information required will be included in the respective ten-
tative specifications monographs (FAO JECFA Monographs 14 (2013)). The
tentative specifications will be withdrawn unless the requested information
is received by the end of 2014.
3.2.4 Food additives containing phosphates: Analytical methods for the

determination of phosphorus and revision of specifications
At its seventy-sixth meeting (Annex 1, reference 211), the Committee intro-
duced an ICP-AES method while preparing the specifications for magne-
sium dihydrogen diphosphate, as the titrimetric and gravimetric methods
incorporated in Volume 4 of the Combined Compendium of Food Additive
Specifications (Annex 1, reference 180) were not reliable for the assay. The
Committee requested information on the suitability of the ICP-AES method
for the assay of other phosphate additives. The Committee, at its current
meeting, agreed to publish the method in FAO JECFA Monographs 14 (2013)
and updated the online version of Volume 4 of the Combined Compendium of
Food Additive Specifications (Annex 1, reference 180) accordingly.
The Committee noted that the general method used for the determination of
cyclic phosphates in polyphosphates, as published in Volume 4 of the Com-
bined Compendium of Food Additive Specifications (Annex 1, reference 180),
is lengthy and uses perchloric acid, which may not be permitted in current
laboratory safety protocols. The Committee recommends that the method
for the determination of cyclic phosphates be reviewed at a future meeting.
34
The Committee also noted that the specifications for some of the phosphate
additives need updating and recommends placing these additives on the
agenda at a future meeting.
3.2.5 Mineral oil (medium viscosity)
The Committee at its seventy-sixth meeting (Annex 1, reference 211) drafted

specifications for mineral oil (medium viscosity) from an existing monograph.
In the call for data for the present meeting, the Committee requested informa-
tion for the revision of provisions and methods of analysis in the specifications.
In response to information received, the Committee agreed to replace the gas
chromatographic method using a packed column for the determination of car-
bon number at 5% distillation with the proposed gas chromatographic method
using a wide-bore column. The replacement method was introduced into the
specifications and will be published in FAO JECFA Monographs 14 (2013).
The method will be added to the online version of Volume 4 of the Combined
Compendium of Food Additive Specifications (Annex 1, reference 180).
3.2.6 Modified starches
The specifications for modified starches cover 16 different modified starches,

including starch sodium octenyl succinate. Modified starches were on the
agenda of the present meeting due to a minor inconsistency in the specifi-
cations of starch sodium octenyl succinate. One of the provisions of starch
sodium octenyl succinate concerns the percentage of octenyl succinate,
whereas the associated analytical method provides the degree of substitu-
tion. The analytical method was therefore revised to calculate the percent-
age of octenyl succinate groups. The revised method will be published in
FAO JECFA Monographs 14 (2013), and the online version will be amended
accordingly.
3.2.7 Paprika extract
The sixty-ninth meeting of the Committee (Annex 1, reference 190) prepared

tentative specifications for paprika extract and required additional informa-
tion in order to be able to remove the tentative status. At the present meet-
ing, the Committee considered the detailed batch information received on
composition, levels of capsaicinoids and levels of arsenic in paprika extract.
The Committee considered the major coloured components and decided to
remove capsorubin from the specifications because it appears to be a minor
component of the article in commerce. The Committee also considered the
levels of capsaicinoids in the samples in commerce and set a maximum
content of 200 mg/kg; maximum arsenic levels were lowered to 1 mg/kg.
The Committee also noted that paprika extract was not soluble in water
and accepted the proposal to use refined soya bean oil as the carrier for the
35
headspace gas chromatographic determination of the residual solvents. The
specifications were revised, and the tentative status was removed. The Chem-
ical and Technical Assessment prepared at the sixty-ninth meeting was modi-
fied to include the composition of commercial preparations.
3.2.8 3-Phytase from Aspergillus niger expressed in Aspergillus niger
The Committee at its seventy-sixth meeting (Annex 1, reference 211) had

prepared the specifications for 3-phytase from Aspergillus niger expressed in
A. niger. CCFA at its Forty-fifth Session (2) requested some minor changes
to the analytical method to measure the activity of 3-phytase. The Committee
revised the specifications to incorporate the proposed changes.
3.2.9 Potassium aluminium silicate
Potassium aluminium silicate was on the agenda of the present meeting for
the purpose of revising the tentative specifications. At its seventy-fourth
meeting (Annex 1, reference 205), the Committee prepared new tentative
specifications for potassium aluminium silicate (PAS). PAS was referred
to the Committee for evaluation as a carrier substrate for pearlescent pig-
ments made with titanium dioxide and/or iron oxide. Information submitted
indicated that PAS was not intended to be placed on the market as an addi-
tive itself; rather, it was only to be used as a carrier substrate as part of the
pearlescent pigments.
At the seventy-fourth meeting, the Committee requested information on the
preparation and purification of PAS, methods of identification, particle size
distribution and inorganic impurities. The Committee also requested infor-
mation on the suitability of the proposed ICP-AES method for the assay. At
the present meeting, the Committee received sufficient information to revise
the specifications. An identification test for aluminium and silicon based
on alkali fusion followed by ICP-AES analysis was introduced. Individual
specifications for metallic impurities were grouped together under the test
“Impurities soluble in 0.5 M hydrochloric acid”. The assay method based on
alkali fusion followed by ICP-AES analysis was retained.
PAS was considered for evaluation as an anticaking agent at the twenty-
eighth and twenty-ninth meetings of the Committee (Annex 1, references 66
and 70). At that time, no information about the manufacture or use of PAS
was provided, and therefore no ADI or specifications could be established.
While no additional information regarding the use of PAS as an anticaking
agent was submitted to the current meeting, the Committee noted that the
Codex Class Names and the International Numbering System for Food Addi-
tives (14) includes PAS (INS No. 555) with the functional class of anticaking
agent. The Committee was aware of possible uses of PAS as an anticaking
36
agent in certain cheese (sliced, cut, shredded or grated cheese) and cocoa
products (cocoa–sugar mixture). Therefore, the functional use of anticaking
agent was added to the specifications monograph for PAS.
The specifications were revised, and the tentative status was removed. A
Chemical and Technical Assessment was prepared.
3.2.10 Potassium aluminium silicate–based pearlescent pigments
The Committee, at its seventy-fourth meeting (Annex 1, reference 205), pre-

pared new tentative specifications for potassium aluminium silicate–based
pearlescent pigments (PAS-BPP) based on information received regarding
the use of PAS as a carrier substrate for titanium dioxide and/or iron oxide.
PAS-BPP are used as colours and are produced by the deposition of titanium
and/or iron salts on PAS followed by calcination at high temperatures. The
resulting pigment consists of PAS coated with titanium dioxide or iron oxide
or a mixture of titanium dioxide and iron oxide.
At its seventy-fourth meeting, the Committee requested information on the
manufacture, stability, particle size distribution and inorganic impurities of
PAS-BPP. The Committee also requested information on the suitability of
the proposed ICP-AES method for the assay.
At the present meeting, PAS-BPP was on the agenda for the purpose of the
revision of tentative specifications. Sufficient information was received to
revise the specifications. The Committee decided to split the combined tenta-
tive specifications prepared at the seventy-fourth meeting into three separate
specifications for PAS-BPP based on the type of material deposited on PAS.
The three specifications are defined as follows: 1) PAS-BPP, Type I: PAS
coated with titanium dioxide only; 2) PAS-BPP, Type II: PAS coated with iron
oxide only; and 3) PAS-BPP, Type III: PAS coated with both titanium dioxide
and iron oxide. An identification test for titanium and/or iron was introduced
based on alkali fusion followed by ICP-AES analysis. Individual specifica-
tions for metallic impurities were grouped together under the test “Impurities
soluble in 0.5 M hydrochloric acid”. The assay method based on alkali fusion
followed by ICP-AES analysis was retained. However, specifications for pH
and particle size, present in the tentative combined specifications monograph
prepared at the seventy-fourth meeting, were removed, as the Committee
considers that these tests could be removed without affecting the identifica-
tion and quality features of the specifications. A general statement under
the definition section in all three monographs was included indicating that
particles of PAS-BPP smaller than 100 nm should not be present.
Although a method of assay was included for all three monographs, an assay
value was not provided, as each of the three PAS-BPP monographs encom-
passes a wide range of pearlescent pigments for each type with different
37
contents of PAS, titanium dioxide and/or iron oxide based on the desired
properties of the pearlescent pigment. As a result, a value of “as labelled”
was provided under the assay to allow for the variability expected for the
multiple types of available pearlescent pigments.
The specifications were revised, and the tentative status was removed. A
Chemical and Technical Assessment was prepared.
38
4. Contaminants
4.1 admium: Assessment of exposure from cocoa and cocoa

C
products
Explanation
Cadmium was evaluated by the Committee at its sixteenth, thirty-third, forty-
first, fifty-fifth, sixty-first, sixty-fourth and seventy-third meetings (Annex 1,
references 30, 83, 107, 149, 166, 176 and 202). At the sixty-first and sixty-
fourth meetings, the Committee noted that the estimated total mean popula-
tion dietary exposure to cadmium from all foods, derived from per capita
data from the five GEMS/Food regional diets, ranged from 40% to 60% of
the provisional tolerable weekly intake (PTWI) applicable at that time, 7 µg/
kg bw. The seven commodity groups that contributed significantly to total
dietary exposure to cadmium were rice, wheat, root vegetables, tuber veg-
etables, leafy vegetables, other vegetables and molluscs (40–85% of the total
dietary exposure to cadmium across the five regional diets).
At its seventy-third meeting, the Committee re-evaluated cadmium and
established a provisional tolerable monthly intake (PTMI) of 25 µg/kg bw,
reflecting the long half-life of cadmium in humans. The estimates of mean
dietary exposure to cadmium from all foods reported from national estimates
for adults ranged from 2.2 to 12 µg/kg bw per month, or 9–48% of the PTMI;
for European children up to 12 years of age, the mean dietary exposure to
cadmium was 11.9 µg/kg bw per month (47% of the PTMI). High-percentile
dietary exposures to cadmium for adults were reported to range from 6.9
to 12.1 µg/kg bw per month (28–48% of the PTMI), and for children aged
0.5–12 years, from 20.4 to 22.0 µg/kg bw per month (82–88% of the PTMI).
Cadmium occurrence data and consumption of foods containing cocoa and
its derivatives were included in these estimates.
CCCF, at its Sixth Session (8), requested that the Committee conduct an
The Committee considered the exposure to cadmium from foods containing
cocoa and its derivatives in the context of overall dietary exposure as reported
at the seventy-third meeting of the Committee.
39
Occurrence data for cadmium in cocoa and cocoa products
The Committee received occurrence data on cadmium in cocoa and cocoa
products from 13 countries (Australia, Czech Republic, Denmark, Ecuador,
Estonia, France, Germany, New Zealand, Romania, Singapore, Slovakia, Swe-
den and the USA). In total, 3919 individual samples collected from 2002 to
2011 were analysed, the majority of which were reported for products available
in the European region. Aggregated mean and median cadmium occurrence
data were submitted from Australia, Ecuador, Singapore and New Zealand,
with information on sample size, but these were not used in the dietary expo-
sure assessment. The Committee classified the submitted data using five
GEMS/Food identifiers: cocoa bean, cocoa powder, cocoa mass, cocoa bev-
erage and other cocoa products (including chocolate). Fifty per cent of the
samples were for other cocoa products, and 33% were for cocoa powder. Of
the total number of samples, 452 were below the limit of quantification (LOQ).
These data were assigned a value of the LOQ, as there was no significant dif-
ference between estimates made using an upper-bound (replacing samples
below the LOQ by the LOQ) or a lower-bound (replacing samples below the
LOQ by 0) occurrence value. The Committee noted that no occurrence data for
cocoa butter were submitted. The occurrence data are summarized in Tables 1
and 2. The guidelines for conducting international dietary exposure assess-
ments for contaminants in foods (9) recommend that when the distribution of
contaminant occurrence data is skewed, the median or geometric mean, rather
than the arithmetic mean, should be used in the dietary exposure estimate. For
cadmium occurrence data where there are a small number of non-detect values
(<LOQ), the use of the geometric mean was considered by the Committee to
be appropriate. As cocoa beans and cocoa mass are not consumed without
further processing, these data were not used in the national dietary exposure
assessments.

International estimates
The guidelines for conducting international dietary exposure assessments for
contaminants in foods using GEMS/Food diets recommend that per capita
data for each Cluster Diet be matched with average contaminant values for
foods containing the food chemical of interest derived as summary values
from occurrence data for the relevant countries in that cluster (9). However, in
this case, cocoa is grown in a restricted area in the world, such that individual
products made from cocoa and its derivatives would tend to have the same
cadmium levels wherever they are offered for purchase. The Committee there-
fore decided to use the summary occurrence data for cocoa mass, as given
in Table 2, to best represent products containing cocoa and its derivatives
at the raw commodity level for the international dietary exposure estimates,
40
Table 1
Summary of cadmium occurrence data for cocoa and cocoa products
Cocoa N Minimum Maximum N > 100 N > 300 N > 500 N > 1000
product (total) concentration concentration µg/kg µg/kg µg/kg µg/kg
(µg/kg) (µg/kg) (%) (%) (%) (%)
Cocoa 451 ND 5239 392 324 245 119
bean (86.9%) (71.8%) (54.3%) (26.4%)
Cocoa 137 ND 290 13 0 0 0
beverage (0.0%)
Cocoa 85 15 593.8 36 6 4 0
mass (37.9%) (6.3%) (4.2%)
Cocoa 1292 ND 1910 669 55 21 6
powder (47.4%) (3.9%) (1.5%) (0.5%)
Other 1954 ND 1073 408 78 7 1
cocoa (20.8%) (4.0%) (0.4%) (0.05%)
products
(including
chocolate)
ND, not detected
Table 2
Summary of statistical descriptors for cadmium occurrence data
Cocoa product Concentration (µg/kg)
Mean Geometric mean Median 97.5th percentile

Cocoa bean 751 467 570 2190
Cocoa beverage 35 22 21 160
Cocoa mass 136 103 88 537
Cocoa powder 130 86 130 430
Other cocoa 76 34 32 361
products (including
chocolate)
as cocoa beverage, cocoa powder and other cocoa products are made from
cocoa mass exported from the producing countries. Per capita food amounts
for cocoa and its derivatives ranged from 0.1 to 7.5 g/day across the 17 Clus-
ter Diets (see Annex 3 for a list of countries included in each of the 17 Cluster
Diets). The geometric mean of the occurrence levels for cocoa mass was mul-
tiplied by the corresponding per capita figure to estimate mean population
dietary exposure to cadmium from cocoa products for each cluster of coun-
tries. These estimates were extrapolated to a monthly basis by multiplying the
daily exposures by 30, then considered relative to the PTMI.
41
The estimates of mean population dietary exposure to cadmium from cocoa
and its derivatives ranged from 0.005 µg/kg bw per month (Cluster 13) to
0.39 µg/kg bw per month (Cluster 7), assuming a 60 kg body weight, which
equated to 0.2–1.6% of the PTMI.
National estimates
The Committee considered a number of exposure calculation scenarios for
preparing the national estimates of dietary exposure to cadmium. These
included combining weighted mean and high-percentile food consumption
data with the geometric mean or high-percentile occurrence data. Two sce-
narios were selected from the options considered to best represent dietary
exposure to cadmium from products containing cocoa and its derivatives:
mean dietary exposures for consumers only were estimated by combining
the geometric mean occurrence data for cadmium for cocoa beverages, cocoa
powder and other cocoa products with the relevant mean food consumption
data for consumers; and 97.5th percentile dietary exposures for consumers
only were estimated by combining geometric mean occurrence data for cad-
mium for cocoa beverages, cocoa powder and other cocoa products with the
relevant 97.5th percentile food consumption data for consumers. The Com-
mittee considered that it was not appropriate to combine the 97.5th percen-
tile food consumption data for consumers only with the 97.5th percentile
occurrence data for a chronic dietary exposure estimate.
The Committee used summary food consumption data derived from indi-
vidual records from a total of 36 different surveys on national consumption
submitted by Brazil, China and the European Food Safety Authority (EFSA)
for the national dietary exposure estimates. For the European data (22 coun-
tries, 34 surveys), the consumption data for cocoa beverages, cocoa powder
and other cocoa products for the whole population (mean amount) and con-
sumers only (mean and 97.5th percentile food consumption amounts) were
combined with the appropriate geometric mean occurrence data to prepare
the dietary exposure estimates for six age groupings taken from the EFSA
data set: toddlers (2–6 years of age), other children (7–11 years of age), ado-
lescents (12–19 years of age), adults (20–65 years of age), elderly (65–75
years of age) and very elderly (>75 years of age). Dietary exposures for the
general population and women of childbearing age in Brazil and China, as
well as for children in China, were also estimated. National dietary exposure
estimates for cadmium from cocoa beverages, cocoa powder and other cocoa
products are summarized in Table 3, with data presented on the mean dietary
exposure for the whole population, for consumers of each product only and
for the 97.5th percentile of exposure for consumers of each product.
Estimated mean dietary exposures to cadmium for the whole population
across different age groups from cocoa beverages ranged from 0.02 to
42
Table 3
Dietary exposure estimates for cocoa derivatives
Cocoa Country/ Age group Dietary exposure (µg/kg bw per month)

product region
Whole Consumers only
population
Mean 97.5th percentile
Cocoa EU Infants — — —
beverages Toddlersa — 2.0 —
Other children 0.058 2.3 11.9
Adolescents 0.138 1.8 6.2
Adults 0.037 1.2 5.2
Elderly 0.020 1.5 4.7
Very elderly 0.017 1.1 2.8
Cocoa EU Infantsa — 0.4 —
powder Toddlers 0.035 0.6 5.1
Adults 0.006 0.1 2.2
Elderly 0.004 0.1 0.6
China General population 0.001 1.6 4.5
Childrenb — — —
Women of childbearing age 0.002 2.3 8.8
Brazil General population 0.118 1.0 3.2
Other EU Infants a
0.006 0.8 1.5
cocoa Toddlers 0.302 1.0 3.7
products
Adults 0.109 0.3 3.7
Elderly 0.052 0.2 1.1
China General population 0.001 0.5 1.8
Children 0.005 1.1 7.8
Brazil General population 0.051 0.9 4.4
a
Number of consumers <11.
b
One consumer reported.
43
0.14 µg/kg bw per month (0.08–0.6% of the PTMI); from cocoa powder,
from 0.001 to 0.13 µg/kg bw per month (0.004–0.5% of the PTMI); and from
other cocoa products, from 0.001 to 0.46 µg/kg bw per month (0.004–1.8%
of the PTMI). Estimated mean dietary exposures across different population
age groups for consumers of cocoa beverages ranged from 1.1 to 2.3 µg/
kg bw per month (4–9% of the PTMI); for consumers of cocoa powder,
from 0.1 to 2.3 µg/kg bw per month (0.4–9% of the PTMI); and for consum-
ers of other cocoa products, from 0.2 to 1.1 µg/kg bw per month (0.8–4%
of the PTMI). Estimated 97.5th percentile dietary exposures to cadmium
across different population age groups for consumers only of cocoa bever-
ages ranged from 2.8 to 11.9 µg/kg bw per month (11–48% of the PTMI);
for consumers only of cocoa powder, from 0.6 to 12.0 µg/kg bw per month
(2–48% of the PTMI); and for consumers only of other cocoa products, from
1.1 to 7.8 µg/kg bw per month (4–31% of the PTMI).
Evaluation
The estimates of mean population dietary exposure to cadmium from prod-
ucts containing cocoa and its derivatives for the 17 GEMS/Food Cluster Diets
ranged from 0.005 to 0.39 µg/kg bw per month, which equated to 0.02–1.6%
of the PTMI. This represents an estimate of mean dietary exposure to cocoa
and its derivatives for the whole population. Similar mean population cad-
mium dietary exposures for individual cocoa products were estimated from
national data, ranging from 0.001 to 0.46 µg/kg bw per month (0.004–1.8%
of the PTMI).
On a national level, it was also possible to estimate dietary exposures for con-
sumers of cocoa products (cocoa beverages, cocoa powder and other cocoa
products). As expected, due to the smaller population of consumers of each
product, these were higher than the population mean dietary exposures. Esti-
mated mean and 97.5th percentile cadmium dietary exposures for consumers
of cocoa beverages and cocoa powder were higher than the corresponding
exposures for consumers of other cocoa products.
The Committee assessed the potential dietary exposure to cadmium for high
consumers of products containing cocoa and its derivatives in addition to
cadmium derived from other foods by adding the highest 97.5th percentile
dietary exposure estimate for adults and children out of the three cocoa food
groups for any of the countries considered to the mean population dietary
exposure estimate for cadmium for adults and children from the whole diet,
as previously estimated at the seventy-third meeting. For adults, the total
cadmium dietary exposure for a high consumer of products containing cocoa
and its derivatives was estimated to be 7.4–17.2 µg/kg bw per month (2.2–
12 µg/kg bw per month for mean exposure from all foods plus 5.2 µg/kg bw
44
per month from the highest 97.5th percentile exposure from cocoa bever-
ages), or 30–69% of the PTMI. For children aged 0.5–12 years, the total
cadmium dietary exposure for a high consumer of products containing cocoa
and its derivatives was estimated to be 23.9 µg/kg bw per month (11.9 µg/kg
bw per month for mean exposure from all foods plus 12 µg/kg bw per month
for the highest 97.5th percentile exposure from cocoa powder), or 96% of
the PTMI. The Committee noted that this total cadmium dietary exposure for
high consumers of cocoa and cocoa products was likely to be overestimated,
because the estimate for dietary exposure to cadmium from the whole diet
also included a contribution from products containing cocoa and its deriva-
tives. The Committee did not consider contributions from products contain-
ing cocoa and its derivatives to total cadmium exposure for high consumers
of these products to be of concern.
No addendum to the toxicological monograph was prepared.
Detailed information on cadmium occurrence data and national food
consumption data used in the evaluation are available on the JECFA web
site5.
5
http://www.who.int/foodsafety/chem/jecfa/publications/reports/en/index.html
45
5. Future work and requests for data
Advantame
New tentative specifications were prepared, pending the submission of infor-
mation, by the end of 2015, on:
• the suitability of the headspace gas chromatographic method (using ap-
propriate dissolution solvent) for determination of residual solvents,
published in Volume 4 of the Combined Compendium of Food Addi-
tive Specifications, and data, in a minimum of five batches, using the
method;
• an alternative or improved high-performance liquid chromatographic
method for the assay of advantame and advantame acid using a standard
curve;
• additional data and analytical methods for the determination of palladium
and platinum;
• information on the purity and availability of the commercial reference
standards used in the assay of advantame and advantame acid.
nalytical method for the determination of carbon number at

A
5% distillation point
The Committee recommended that a note be included in Volume 4 of the
Combined Compendium of Food Additive Specifications to indicate the avail-
ability of a newer method. The Committee further recommended that the
suitability of this method for use in the analysis of similar substances be
evaluated at a future meeting.
nalytical method for the determination of residual solvents by

A
headspace gas chromatography
The Committee recommended that the issue of the suitability of dissolu-
tion solvents for the determination of residual solvents in food additives be
investigated at a future meeting.
47
nnatto extracts (solvent-extracted bixin and solvent-extracted
A
norbixin)
The Committee recommended that manufacturers supply residual solvent data
from at least five batches of each of the solvent-extracted bixin and norbixin
products to support the possible revision of the provision for residual solvents.
To evaluate the suitability of the method for the determination of residual
solvents in annatto extracts dissolved in dimethyl formamide, the Committee
also recommended that manufacturers provide results from the analysis of
samples of solvent-extracted bixin and norbixin products using this method as
well as the general method for the determination of residual solvents published
in Volume 4 of the Combined Compendium of Food Additive Specifications.
Benzoe tonkinensis
The tentative specifications will be withdrawn if the complete data on the
composition of the ethanolic extract and microbiological contaminants are
not received by the end of 2013.
Food additives containing aluminium and/or silicon

Specifications were made tentative pending the submission of information
on composition; methods of manufacture; data on loss on drying and loss on
ignition; impurities (lead, cadmium, arsenic and mercury) soluble in hydro-
chloric acid (0.5 mol/l); and suitability of the proposed ICP-AES method
for assay, as well as data on the assay. Details on information required are
included in the respective tentative specifications monographs. The tentative
specifications will be withdrawn unless the requested information is received
by the end of 2014.
Glycerol ester of gum rosin

The specifications were maintained as tentative pending the submission of
additional information by the end of 2014. Additional data are requested
to characterize GEGR in commerce in relation to the composition of 1) the
refined gum rosin currently used as the source rosin with regard to the levels
(%) of resin acids and neutrals, 2) the glycerol ester of gum rosin with regard
to the levels (%) of a) glycerol esters, b) free resin acids and c) neutrals
and 3) the total glycerol esters of resin acids with regard to the levels (%)
of a) glycerol monoesters and b) the sum of glycerol diesters and triesters
(assay). Validated methods for the determination of the substances consid-
ered in the specifications are also required.
Octenyl succinic acid modified gum arabic

The Committee noted that ongoing studies on the stability of OSA modified
gum arabic in food may provide further information on its chemical state in
48
food and aqueous solutions, which could help to explain the contradictory
results of the hydrolysis study submitted to the Committee at the present
meeting. The Committee decided to retain the temporary ADI “not speci-
fied” pending submission of additional data on the stability of OSA modified
gum arabic in food by the end of 2013.
The Committee noted that the purity test of degree of esterification in the
current specifications should be replaced by the degree of substitution and
requested information for an analytical method to measure the degree of
substitution and results of the analysis of at least five commercially available
batches. The specifications were made tentative pending submission of these
data by the end of 2013.
hosphates: Analytical methods for the determination of

P
phosphorus and revision of specifications
The Committee noted that the general method used for the determination
of cyclic phosphates in polyphosphates, as published in Volume 4 of the
Combined Compendium of Food Additive Specifications, is lengthy and uses
perchloric acid, which may not be permitted in current laboratory safety pro-
tocols. The Committee recommends that the method for the determination of
cyclic phosphates be reviewed at a future meeting.
The Committee also noted that the specifications for some of the phosphate
additives need updating and recommends placing these additives on the
agenda at a future meeting.
49
6. Recommendations
nnatto extracts (solvent-extracted bixin and solvent-extracted

A
norbixin)
The Committee recommends that manufacturers supply residual solvent data
from at least five batches of each of the solvent-extracted bixin and nor-
bixin products to support the possible revision of the provision for residual
solvents.
In order to evaluate the suitability of a method for the determination of
residual solvents in annatto extracts dissolved in dimethyl formamide for
the analysis of solvent-extracted bixin and norbixin products, the Committee
recommends that manufacturers provide results from the analysis of samples
of solvent-extracted bixin and norbixin products using this method as well
as the general method for the determination of residual solvents published in
Volume 4 of the Combined Compendium of Food Additive Specifications for
the analysis of solvent-extracted bixin and norbixin products.
Requirements for submission of analytical methods

In order to assess and ensure the accuracy and reliability of the data submit-
ted, the Committee recommends the use of methods that are appropriately
validated. It also recommends that in relevant cases, the detailed analytical
method be provided, together with validation data, in response to specific
JECFA calls for data.
51
Acknowledgements
The Committee wishes to thank Ms M. Sheffer, Ottawa, Canada, for her

assistance in the preparation of the report. The Committee also expresses
its appreciation to Ms Clélia Bianchi, Intern, Department of Food Safety
and Zoonoses, WHO, Geneva, for her presentation on the new GEMS/Food
Cluster Diets.
53
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56
Annex 1
Reports and other documents
resulting from previous meetings
of the Joint FAO/WHO Expert
Committee on Food Additives
1. General principles governing the use of food additives (First report

of the Joint FAO/WHO Expert Committee on Food Additives). FAO
Nutrition Meetings Report Series, No. 15, 1957; WHO Technical Report
Series, No. 129, 1957 (out of print).
2. Procedures for the testing of intentional food additives to establish their
safety for use (Second report of the Joint FAO/WHO Expert Committee
on Food Additives). FAO Nutrition Meetings Report Series, No. 17,
1958; WHO Technical Report Series, No. 144, 1958 (out of print).
3. Specifications for identity and purity of food additives (antimicrobial
preservatives and antioxidants) (Third report of the Joint FAO/WHO
Expert Committee on Food Additives). These specifications were
subsequently revised and published as Specifications for identity
and purity of food additives, Vol. I. Antimicrobial preservatives and
antioxidants, Rome, Food and Agriculture Organization of the United
Nations, 1962 (out of print).
4. Specifications for identity and purity of food additives (food colours)
(Fourth report of the Joint FAO/WHO Expert Committee on Food
Additives). These specifications were subsequently revised and
published as Specifications for identity and purity of food additives,
Vol. II. Food colours, Rome, Food and Agriculture Organization of the
United Nations, 1963 (out of print).
5. Evaluation of the carcinogenic hazards of food additives (Fifth report
Nutrition Meetings Report Series, No. 29, 1961; WHO Technical Report
Series, No. 220, 1961 (out of print).
6. Evaluation of the toxicity of a number of antimicrobials and antioxidants
(Sixth report of the Joint FAO/WHO Expert Committee on Food
57
Additives). FAO Nutrition Meetings Report Series, No. 31, 1962; WHO
Technical Report Series, No. 228, 1962 (out of print).
7. Specifications for the identity and purity of food additives and their
toxicological evaluation: emulsifiers, stabilizers, bleaching and maturing
agents (Seventh report of the Joint FAO/WHO Expert Committee on
Food Additives). FAO Nutrition Meetings Series, No. 35, 1964; WHO
toxicological evaluation: food colours and some antimicrobials and
antioxidants (Eighth report of the Joint FAO/WHO Expert Committee
on Food Additives). FAO Nutrition Meetings Series, No. 38, 1965; WHO
9. Specifications for identity and purity and toxicological evaluation of
some antimicrobials and antioxidants. FAO Nutrition Meetings Report
Series, No. 38A, 1965; WHO/Food Add/24.65 (out of print).
10. Specifications for identity and purity and toxicological evaluation of food
colours. FAO Nutrition Meetings Report Series, No. 38B, 1966; WHO/
Food Add/66.25.
toxicological evaluation: some antimicrobials, antioxidants, emulsifiers,
stabilizers, flour treatment agents, acids, and bases (Ninth report of the
Joint FAO/WHO Expert Committee on Food Additives). FAO Nutrition
Meetings Series, No. 40, 1966; WHO Technical Report Series, No. 339,
1966 (out of print).
12. Toxicological evaluation of some antimicrobials, antioxidants,
emulsifiers, stabilizers, flour treatment agents, acids, and bases. FAO
Nutrition Meetings Report Series, No. 40A, B, C; WHO/Food Add/67.29.
toxicological evaluation: some emulsifiers and stabilizers and certain
other substances (Tenth report of the Joint FAO/WHO Expert Committee
on Food Additives). FAO Nutrition Meetings Series, No. 43, 1967; WHO
Technical Report Series, No. 373, 1967.
toxicological evaluation: some flavouring substances and non nutritive
sweetening agents (Eleventh report of the Joint FAO/WHO Expert
Committee on Food Additives). FAO Nutrition Meetings Series, No. 44,
1968; WHO Technical Report Series, No. 383, 1968.
15. Toxicological evaluation of some flavouring substances and non
nutritive sweetening agents. FAO Nutrition Meetings Report Series, No.
44A, 1968; WHO/Food Add/68.33.
16. Specifications and criteria for identity and purity of some flavouring
substances and non-nutritive sweetening agents. FAO Nutrition
Meetings Report Series, No. 44B, 1969; WHO/Food Add/69.31.
58
toxicological evaluation: some antibiotics (Twelfth report of the Joint FAO/
WHO Expert Committee on Food Additives). FAO Nutrition Meetings
Series, No. 45, 1969; WHO Technical Report Series, No. 430, 1969.
18. Specifications for the identity and purity of some antibiotics. FAO
Nutrition Meetings Series, No. 45A, 1969; WHO/Food Add/69.34.
toxicological evaluation: some food colours, emulsifiers, stabilizers,
anticaking agents, and certain other substances (Thirteenth report
Nutrition Meetings Series, No. 46, 1970; WHO Technical Report Series,
No. 445, 1970.
20. Toxicological evaluation of some food colours, emulsifiers, stabilizers,
anticaking agents, and certain other substances. FAO Nutrition
Meetings Report Series, No. 46A, 1970; WHO/Food Add/70.36.
21. Specifications for the identity and purity of some food colours,
emulsifiers, stabilizers, anticaking agents, and certain other food
additives. FAO Nutrition Meetings Report Series, No. 46B, 1970; WHO/
Food Add/70.37.
22. Evaluation of food additives: specifications for the identity and purity
of food additives and their toxicological evaluation: some extraction
solvents and certain other substances; and a review of the technological
efficacy of some antimicrobial agents (Fourteenth report of the Joint
FAO/WHO Expert Committee on Food Additives). FAO Nutrition
1971.
23. Toxicological evaluation of some extraction solvents and certain other
substances. FAO Nutrition Meetings Report Series, No. 48A, 1971;
WHO/Food Add/70.39.
24. Specifications for the identity and purity of some extraction solvents
and certain other substances. FAO Nutrition Meetings Report Series,
No. 48B, 1971; WHO/Food Add/70.40.
25. A review of the technological efficacy of some antimicrobial agents.
FAO Nutrition Meetings Report Series, No. 48C, 1971; WHO/Food
Add/70.41.
26. Evaluation of food additives: some enzymes, modified starches, and
certain other substances: Toxicological evaluations and specifications
and a review of the technological efficacy of some antioxidants (Fifteenth
report of the Joint FAO/WHO Expert Committee on Food Additives).
FAO Nutrition Meetings Series, No. 50, 1972; WHO Technical Report
Series, No. 488, 1972.
27. Toxicological evaluation of some enzymes, modified starches, and
certain other substances. FAO Nutrition Meetings Report Series, No.
50A, 1972; WHO Food Additives Series, No. 1, 1972.
59
28. Specifications for the identity and purity of some enzymes and certain
other substances. FAO Nutrition Meetings Report Series, No. 50B,
1972; WHO Food Additives Series, No. 2, 1972.
29. A review of the technological efficacy of some antioxidants and
synergists. FAO Nutrition Meetings Report Series, No. 50C, 1972;
30. Evaluation of certain food additives and the contaminants mercury,
lead, and cadmium (Sixteenth report of the Joint FAO/WHO Expert
Committee on Food Additives). FAO Nutrition Meetings Series, No. 51,
1972; WHO Technical Report Series, No. 505, 1972, and corrigendum.
31. Evaluation of mercury, lead, cadmium and the food additives amaranth,
diethylpyrocarbamate, and octyl gallate. FAO Nutrition Meetings Report
Series, No. 51A, 1972; WHO Food Additives Series, No. 4, 1972.
32. Toxicological evaluation of certain food additives with a review of
general principles and of specifications (Seventeenth report of the
Joint FAO/WHO Expert Committee on Food Additives). FAO Nutrition
1974, and corrigendum (out of print).
33. Toxicological evaluation of some food additives including anticaking
agents, antimicrobials, antioxidants, emulsifiers, and thickening agents.
FAO Nutrition Meetings Report Series, No. 53A, 1974; WHO Food
Additives Series, No. 5, 1974.
34. Specifications for identity and purity of thickening agents, anticaking
agents, antimicrobials, antioxidants and emulsifiers. FAO Food and
Nutrition Paper, No. 4, 1978.
35. Evaluation of certain food additives (Eighteenth report of the Joint FAO/
WHO Expert Committee on Food Additives). FAO Nutrition Meetings
Series, No. 54, 1974; WHO Technical Report Series, No. 557, 1974,
and corrigendum.
36. Toxicological evaluation of some food colours, enzymes, flavour
enhancers, thickening agents, and certain other food additives. FAO
Nutrition Meetings Report Series, No. 54A, 1975; WHO Food Additives
Series, No. 6, 1975.
37. Specifications for the identity and purity of some food colours, enhancers,
thickening agents, and certain food additives. FAO Nutrition Meetings
Report Series, No. 54B, 1975; WHO Food Additives Series, No. 7, 1975.
38. Evaluation of certain food additives: some food colours, thickening
agents, smoke condensates, and certain other substances. (Nineteenth
FAO Nutrition Meetings Series, No. 55, 1975; WHO Technical Report
Series, No. 576, 1975.
39. Toxicological evaluation of some food colours, thickening agents, and
certain other substances. FAO Nutrition Meetings Report Series, No.
55A, 1975; WHO Food Additives Series, No. 8, 1975.
60
40. Specifications for the identity and purity of certain food additives. FAO
Nutrition Meetings Report Series, No. 55B, 1976; WHO Food Additives
Series, No. 9, 1976.
41. Evaluation of certain food additives (Twentieth report of the Joint FAO/
WHO Expert Committee on Food Additives). FAO Food and Nutrition
1976.
42. Toxicological evaluation of certain food additives. WHO Food Additives
Series, No. 10, 1976.
43. Specifications for the identity and purity of some food additives. FAO
Food and Nutrition Series, No. 1B, 1977; WHO Food Additives Series,
No. 11, 1977.
44. Evaluation of certain food additives (Twenty-first report of the Joint
FAO/WHO Expert Committee on Food Additives). WHO Technical
Report Series, No. 617, 1978.
45. Summary of toxicological data of certain food additives. WHO Food
Additives Series, No. 12, 1977.
46. Specifications for identity and purity of some food additives, including
antioxidant, food colours, thickeners, and others. FAO Nutrition
Meetings Report Series, No. 57, 1977.
47. Evaluation of certain food additives and contaminants (Twenty-second
WHO Technical Report Series, No. 631, 1978.
48. Summary of toxicological data of certain food additives and
contaminants. WHO Food Additives Series, No. 13, 1978.
Food and Nutrition Paper, No. 7, 1978.
50. Evaluation of certain food additives (Twenty-third report of the Joint
Report Series, No. 648, 1980, and corrigenda.
Series, No. 14, 1980.
52. Specifications for identity and purity of food colours, flavouring agents,
and other food additives. FAO Food and Nutrition Paper, No. 12, 1979.
53. Evaluation of certain food additives (Twenty-fourth report of the Joint
Series, No. 15, 1980.
55. Specifications for identity and purity of food additives (sweetening
agents, emulsifying agents, and other food additives). FAO Food and
Nutrition Paper, No. 17, 1980.
61
56. Evaluation of certain food additives (Twenty-fifth report of the Joint FAO/
WHO Expert Committee on Food Additives). WHO Technical Report
Series, No. 669, 1981.
Series, No. 16, 1981.
58. Specifications for identity and purity of food additives (carrier solvents,
emulsifiers and stabilizers, enzyme preparations, flavouring agents,
food colours, sweetening agents, and other food additives). FAO Food
and Nutrition Paper, No. 19, 1981.
59. Evaluation of certain food additives and contaminants (Twenty-sixth
Series, No. 17, 1982.
62. Evaluation of certain food additives and contaminants (Twenty-seventh
WHO Technical Report Series, No. 696, 1983, and corrigenda.
63. Toxicological evaluation of certain food additives and contaminants.
65. Guide to specifications—General notices, general methods,
identification tests, test solutions, and other reference materials. FAO
Food and Nutrition Paper, No. 5, Rev. 1, 1983.
66. Evaluation of certain food additives and contaminants (Twenty-eighth
WHO Technical Report Series, No. 710, 1984, and corrigendum.
68. Specifications for the identity and purity of food colours. FAO Food and
Nutrition Paper, No. 31/1, 1984.
69. Specifications for the identity and purity of food additives. FAO Food
and Nutrition Paper, No. 31/2, 1984.
70. Evaluation of certain food additives and contaminants (Twenty-ninth
WHO Food Additives Series, No. 20. Cambridge University Press, 1987.
62
73. Evaluation of certain food additives and contaminants (Thirtieth report
of the Joint FAO/WHO Expert Committee on Food Additives). WHO
WHO Food Additives Series, No. 21. Cambridge University Press, 1987.
76. Principles for the safety assessment of food additives and contaminants
in food. WHO Environmental Health Criteria, No. 70. Geneva, World
Health Organization, 1987 (out of print). The full text is available
electronically at www.who.int/pcs.
77. Evaluation of certain food additives and contaminants (Thirty-first
Series, No. 22. Cambridge University Press, 1988.
80. Evaluation of certain veterinary drug residues in food (Thirty-second
81. Toxicological evaluation of certain veterinary drug residues in food.
WHO Food Additives Series, No. 23. Cambridge University Press,
1988.
82. Residues of some veterinary drugs in animals and foods. FAO Food
83. Evaluation of certain food additives and contaminants (Thirty-third
WHO Food Additives Series, No. 24. Cambridge University Press,
1989.
85. Evaluation of certain veterinary drug residues in food (Thirty-fourth
88. Evaluation of certain food additives and contaminants (Thirty-fifth
63
90. Specifications for identity and purity of certain food additives. FAO Food
91. Evaluation of certain veterinary drug residues in food (Thirty-sixth
94. Evaluation of certain food additives and contaminants (Thirty-seventh
96. Compendium of food additive specifications (Joint FAO/WHO Expert
Committee on Food Additives (JECFA)). Combined specifications from
1st through the 37th meetings, 1956–1990. Rome, Food and Agriculture
Organization of the United Nations, 1992 (2 volumes).
97. Evaluation of certain veterinary drug residues in food (Thirty-eighth
98. Toxicological evaluation of certain veterinary residues in food. WHO
Food Additives Series, No. 29, 1991.
100. Guide to specifications—General notices, general analytical techniques,
identification tests, test solutions, and other reference materials. FAO
Food and Nutrition Paper, No. 5, Ref. 2, 1991.
101. Evaluation of certain food additives and naturally occurring toxicants
(Thirty-ninth report of the Joint FAO/WHO Expert Committee on Food
Additives). WHO Technical Report Series No. 828, 1992.
102. Toxicological evaluation of certain food additives and naturally occurring
toxicants. WHO Food Additives Series, No. 30, 1993.
103. Compendium of food additive specifications: addendum 1. FAO Food
104. Evaluation of certain veterinary drug residues in food (Fortieth report
64
106. Residues of some veterinary drugs in animals and food. FAO Food and
Nutrition Paper, No. 41/5, 1993.
107. Evaluation of certain food additives and contaminants (Forty-first report
and Nutrition Paper, No. 52, Add. 2, 1993.
110. Evaluation of certain veterinary drug residues in food (Forty-second
113. Evaluation of certain veterinary drug residues in food (Forty-third report
Technical Report Series, No. 855, 1995, and corrigendum.
116. Evaluation of certain food additives and contaminants (Forty-fourth
119. Evaluation of certain veterinary drug residues in food (Forty-fifth report
122. Evaluation of certain food additives and contaminants (Forty-sixth
Series, No. 37, 1996.
65
124. Compendium of food additive specifications, addendum 4. FAO Food
125. Evaluation of certain veterinary drug residues in food (Forty-seventh
128. Evaluation of certain veterinary drug residues in food (Forty-eighth
131. Evaluation of certain food additives and contaminants (Forty-ninth
132. Safety evaluation of certain food additives and contaminants. WHO
134. Evaluation of certain veterinary drug residues in food (Fiftieth report
137. Evaluation of certain food additives (Fifty-first report of the Joint FAO/
Series, No. 891, 2000.
138. Safety evaluation of certain food additives. WHO Food Additives Series,
No. 42, 1999.
140. Evaluation of certain veterinary drug residues in food (Fifty-second
66
143. Evaluation of certain food additives and contaminants (Fifty-third report
146. Evaluation of certain veterinary drug residues in food (Fifty-fourth
149. Evaluation of certain food additives and contaminants (Fifty-fifth report
152. Evaluation of certain mycotoxins in food (Fifty-sixth report of the Joint
153. Safety evaluation of certain mycotoxins in food. WHO Food Additives
Series, No. 47/FAO Food and Nutrition Paper 74, 2001.
154. Evaluation of certain food additives and contaminants (Fifty-seventh
157. Evaluation of certain veterinary drug residues in food (Fifty-eighth
67
160. Evaluation of certain food additives and contaminants (Fifty-ninth
163. Evaluation of certain veterinary drug residues in food (Sixtieth report
166. Evaluation of certain food additives and contaminants (Sixty-first report
169. Evaluation of certain veterinary drug residues in food (Sixty-second
173. Evaluation of certain food additives (Sixty-third report of the Joint FAO/
Series, No. 928, 2005.
No 54, 2005.
and Nutrition Paper, No. 52, Add. 13 (with Errata), 2005.
176. Evaluation of certain food contaminants (Sixty-fourth report of the
Joint FAO/WHO Expert Committee on Food Additives). WHO Technical
177. Safety evaluation of certain contaminants in food. WHO Food Additives
Series, No. 55/FAO Food and Nutrition Paper, No. 82, 2006.
68
178. Evaluation of certain food additives (Sixty-fifth report of the Joint FAO/
Series, No. 934, 2006.
No. 56, 2006.
180. Combined compendium of food additive specifications. FAO JECFA
Monographs 1, Volumes 1–4, 2005, 2006.
181. Evaluation of certain veterinary drug residues in food (Sixty-sixth report
182. Residue evaluation of certain veterinary drugs. FAO JECFA Monographs
2, 2006.
184. Evaluation of certain food additives and contaminants (Sixty-seventh
185. Compendium of food additive specifications. FAO JECFA Monographs
3, 2006.
187. Evaluation of certain food additives and contaminants (Sixty-eighth
4, 2007.
190. Evaluation of certain food additives (Sixty-ninth report of the Joint FAO/
Series, No. 952, 2009.
No. 60, 2009.
5, 2009.
193. Evaluation of certain veterinary drug residues in food (Seventieth
6, 2009.
69
196. Evaluation of certain food additives (Seventy-first report of the Joint
No. 62, 2010.
7, 2009.
199. Evaluation of certain contaminants in food (Seventy-second report
200. Safety evaluation of certain contaminants in food. WHO Food Additives
Series, No. 63/FAO JECFA Monographs 8, 2011.
9, 2010.
202. Evaluation of certain food additives and contaminants (Seventy-third
10, 2010.
205. Evaluation of certain food additives and contaminants (Seventy-fourth
11, 2011.
208. Evaluation of certain veterinary drug residues in food (Seventy-fifth
12, 2012.
211. Evaluation of certain food additives (Seventy-sixth report of the Joint
No. 67, 2012.
13, 2012.
70
Annex 2
Toxicological and dietary exposure
information and information on
specifications
Food additives considered for specifications only
Food additive Specificationsa

Aluminium silicate R, T
Annatto extracts (solvent-extracted bixin) M
Annatto extracts (solvent-extracted norbixin) M
Benzoe tonkinensis M, T
Calcium aluminium silicate R, T
Calcium silicate R, T
Food additives containing phosphates Rb
Mineral oil (medium viscosity) R
Modified starches Rc
Paprika extract Rd
Phytase from Aspergillus niger expressed in Aspergillus niger R
Potassium aluminium silicate Rd
Potassium aluminium silicate–based pearlescent pigments, Type I Ne
Potassium aluminium silicate–based pearlescent pigments, Type II Ne
Potassium aluminium silicate–based pearlescent pigments, Type III Ne
Silicon dioxide, amorphous R, T
Sodium aluminosilicate R, T
M, existing specifications maintained; N, new specifications; R, existing specifications revised; T, tentative
a
specifications.
The inductively coupled plasma – atomic emission spectrophotometric (ICP-AES) method for the assay
b
of phosphate additives was added to the Combined Compendium of Food Additive Specifications.
The method for determination of percentage of octenyl succinate groups in starch sodium octenyl
c
succinate was revised.

The tentative status of the specifications was removed.
d
The existing combined specifications for potassium aluminium silicate–based pearlescent pigments
e
were split into three separate specifications (Type I: coated with titanium oxide only, Type II: coated with
iron oxide only and Type III: coated with both titanium dioxide and iron oxide). The tentative status of the
specifications was removed.
71
ood additives evaluated toxicologically, assessed for dietary exposure and
F
considered for specifications
Food Specificationsa Acceptable daily intakes, other toxicological

additive recommendations and dietary exposure assessment
Advantame N, T The Committee established an acceptable daily
intake (ADI) of 0–5 mg/kg body weight (bw) for
advantame on the basis of a no-observed-adverse-effect
level (NOAEL) of 500 mg/kg bw per day for maternal
toxicity in a developmental toxicity study in rabbits and
application of a 100-fold safety factor to account for
interspecies and intraspecies variability.
The Committee agreed that the ADI also applies to those
individuals with phenylketonuria, as the formation of
phenylalanine from the normal use of advantame would
not be significant in relation to this condition.
Using the proposed maximum use levels and
conservative assumptions, the maximum mean dietary
exposure to advantame would be 1.45 mg/kg bw per day
(29% of the upper bound of the ADI), and the maximum
high-percentile dietary exposure would be 2.16 mg/kg bw
per day (43% of the upper bound of the ADI).
Glucoamylase N Based on its low toxicity and because it is reasonably
from anticipated that dietary exposure would be very low, the
Trichoderma Committee established an ADI “not specified”b for
reesei the glucoamylase enzyme preparation from T. reesei
expressed in expressed in T. reesei used in the applications specified
Trichoderma and in accordance with good manufacturing practice.
reesei
Glycerol ester R, T As the requested two unpublished 90-day oral toxicity
of gum rosin studies on GEGR in rats and complete information
(GEGR) on the composition of GEGR were not submitted, the
Committee withdrew the temporary group ADI of
0–12.5 mg/kg bw for GEGR and glycerol ester of
wood rosin (GEWR) (see below).
Glycerol ester W No data on GETOR were submitted, and the Secretariat
of tall oil rosin was informed that this compound is no longer supported
(GETOR) by the previous data sponsor. Therefore, the Committee
did not evaluate GETOR.
Glycerol ester Rc As the requested data on GEGR were not submitted,
of wood rosin the Committee withdrew the temporary group ADI
(GEWR) of 0–12.5 mg/kg bw for GEGR and GEWR and re-
established the ADI of 0–25 mg/kg bw for GEWR.
72
Food Specificationsa Acceptable daily intakes, other toxicological
additive recommendations and dietary exposure assessment
Nisin R The Committee established an ADI for nisin of 0–2
mg/kg bw on the basis of a NOAEL of 224.7 mg of
nisin per kilogram body weight per day from a 13-week
study in rats and application of a safety factor of 100
to account for interspecies and intraspecies variability.
The Committee did not consider it necessary to use an
additional safety factor to account for the short duration
of the study because no compound-related effects were
observed at any dose in any of the toxicity studies,
including a reproductive toxicity study in rats, and
because ingested nisin is degraded in the upper part of
the intestinal tract, such that systemic exposure to nisin
is not likely to occur.
The highest estimated dietary exposure of 0.07 mg of
nisin per kilogram body weight per day determined at the
current meeting did not exceed the upper bound of the
ADI.
The Committee withdrew the previous ADI of 0–33 000
units of nisin per kilogram body weight established at
the twelfth meeting.
Octenyl R, T The Committee decided to retain the temporary ADI
succinic “not specified”b pending submission of additional data
acid (OSA) on the stability of OSA modified gum arabic in food by
modified gum the end of 2013, which may help to explain contradictory
arabic hydrolysis data.
a
M, existing specifications maintained; N, new specifications; R, existing specifications revised; T, tentative
specifications; W, existing specifications withdrawn.
b
ADI “not specified” is used to refer to a food substance of very low toxicity that, on the basis of the
available data (chemical, biochemical, toxicological and other) and the total dietary exposure to the
substance arising from its use at the levels necessary to achieve the desired effects and from its ac-
ceptable background levels in food, does not, in the opinion of the Committee, represent a hazard to
health. For that reason, and for the reasons stated in the individual evaluations, the establishment of an
ADI expressed in numerical form is not deemed necessary. An additive meeting this criterion must be
used within the bounds of good manufacturing practice—i.e. it should be technologically efficacious and
should be used at the lowest level necessary to achieve this effect, it should not conceal food of inferior
quality or adulterated food, and it should not create a nutritional imbalance.
c
The tentative status of the specifications was removed.
Contaminants
Cadmium: Assessment of exposure from cocoa and cocoa products
The Codex Committee on Contaminants in Foods, at its Sixth Session, requested that
the Committee conduct an assessment of dietary exposure to cadmium from cocoa
and cocoa products.
The estimates of mean population dietary exposure to cadmium from products con-
taining cocoa and its derivatives for the 17 new Global Environment Monitoring
System – Food Contamination Monitoring and Assessment Programme (GEMS/
73
Food) Cluster Diets (see Annex 3) ranged from 0.005 to 0.39 µg/kg bw per month,
which equated to 0.02–1.6% of the provisional tolerable monthly intake (PTMI) of
25 µg/kg bw. Similar mean population cadmium dietary exposures for individual
cocoa products were estimated from national data, ranging from 0.001 to 0.46 µg/kg
bw per month (0.004–1.8% of the PTMI).
The potential dietary exposures to cadmium for high consumers of products contain-
ing cocoa and its derivatives in addition to cadmium derived from other foods were
estimated to be 30–69% of the PTMI for adults and 96% of the PTMI for children
0.5–12 years of age. The Committee noted that this total cadmium dietary exposure
for high consumers of cocoa and cocoa products was likely to be overestimated and
did not consider it to be of concern.
Detailed information on cadmium occurrence data and national food consumption
data used in the evaluation will be available on the JECFA web site.
74
Annex 3
GEMS/Food Cluster Diets 2012
Cluster/country Cluster/country Cluster/country Cluster/country Cluster/country

G01 Togo Nicaragua Democratic Ethiopia
Afghanistan Zambia Panama People’s Gambia
Algeria Peru Republic of Haiti
Azerbaijan G04 Seychelles Korea Kenya
Iraq Antigua and South Africa Guinea Bissau Malawi
Jordan Barbuda Suriname Indonesia Mali
Libya Bahamas Tajikistan Lao People’s Namibia
Mauritania Barbados The former Democratic Niger
Mongolia Brunei Darussalam Yugoslav Republic Nigeria
Morocco French Polynesia Republic of Myanmar Senegal
Occupied Grenada Macedonia Nepal Somalia
Palestian Israel Trinidad and Philippines Sudan
Territory Jamaica Tobago Sierra Leone Swaziland
Pakistan Kuwait Venezuela, Thailand United Republic of
Syrian Arab Netherlands Bolivarian Timor Leste Tanzania
Republic Antilles Republic of Viet Nam Zimbabwe
Tunisia Saint Kitts and
Turkmenistan Nevis G06 G10 G14
Uzbekistan Saint Lucia Armenia Belarus Comoros
Yemen Saint Vincent and Cuba Bulgaria Fiji Islands
the Grenadines Egypt Canada Kiribati
G02 Saudi Arabia Greece Croatia Papua New Guinea
Albania United Arab Iran, Islamic Cyprus Solomon Islands
Bosnia and Emirates Republic of Estonia Sri Lanka
Herzegovina Lebanon Italy Vanuatu
Georgia G05 Turkey Japan
Kazakhstan Argentina Latvia G15
Kyrgyzstan Bolivia, G07 Malta Czech Republic
Montenegro Plurinational Australia New Zealand Denmark
Republic of State of Bermuda Republic of Korea Hungary
Moldova Brazil Finland Russian Ireland
Ukraine Cape Verde France Federation Lithuania
Chile Iceland United States of Portugal
G03 Colombia Luxembourg America Romania
Angola Costa Rica Norway Serbia and
Benin Djibouti Switzerland G11 Montenegro
Burundi Dominican United Kingdom Belgium Slovakia
Cameroon Republic Uruguay Netherlands Slovenia
Congo Ecuador Sweden
Côte d’Ivoire El Salvador G08 G12
Democratic Guatemala Austria Belize G16
Republic of the Guyana Germany Dominica Gabon
Congo Honduras Poland Rwanda
Ghana India Spain G13 Uganda
Guinea Malaysia Botswana
Liberia Maldives G09 Burkina Faso G17
Madagascar Mauritius Bangladesh Central African Samoa
Mozambique Mexico Cambodia Republic Sao Tome and
Paraguay New Caledonia China Chad Principe
75
983
W H O Te c h n i c a l R e p o r t S e r i e s
983

This report represents the conclusions of a Joint FAO/WHO Expert

Committee convened to evaluate the safety of various food additives and
a food contaminant with a view to concluding as to safety concerns and to
preparing specifications for identity and purity.
The first part of the report contains a general discussion of the principles
governing the toxicological evaluation of and assessment of dietary exposure
to food additives. A summary follows of the Committee’s evaluations of
technical, toxicological and dietary exposure data for seven food additives
(advantame; glucoamylase from Trichoderma reesei expressed in Trichoderma
reesei; glycerol ester of gum rosin; glycerol ester of tall oil rosin; glycerol ester
of wood rosin; nisin; and octenyl succinic acid modified gum arabic) and an
food additives and
Specifications for the following food additives were revised: annatto extracts
(solvent-extracted bixin and solvent-extracted norbixin); Benzoe tonkinensis;
contaminants
food additives containing aluminium and/or silicon; mineral oil (medium
viscosity); modified starches; paprika extract; phosphates (analytical methods
for the determination of phosphorus and revision of specifications); 3-phytase Seventy-seventh report of the
from Aspergillus niger expressed in Aspergillus niger; potassium aluminium Joint FAO/WHO Expert Committee on
silicate; and potassium aluminium silicate–based pearlescent pigments.
WHO Technical Report Series

Food Additives
Annexed to the report are tables summarizing the Committee’s
recommendations for dietary exposures to and toxicological evaluations of
the food additives and contaminant considered.
Organization of
the United Nations
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983

Evaluation of certain food additives and contaminants

This report represents the conclusions of a Joint FAO/WHO Expert

WHO Technical Report Series

TRS 983-cover.indd 1 12/16/2013 6:37:00 PM

(WHO technical report series ; no. 983)

1.Food additives - analysis. 2.Food additives - toxicity. 3.Food contaminants - analysis.

ISBN 978 92 4 120983 0 (NLM classification: WA 712)

© World Health Organization 2013

3. Specific food additives 11

5. Future work and requests for data 47

Annex 1 Reports and other documents resulting from previous

Annex 2 Toxicological and dietary exposure information and

Annex 3 GEMS/Food Cluster Diets 2012 75

The Joint FAO/WHO Expert Committee on Food Additives (JECFA) met in

1.1 Declarations of interests

As a result of the recommendations of the first Joint FAO/WHO Conference

2.1 Modification of the agenda

2.2  eport from the Forty-fifth Session of the Codex Committee on

2.3  eport from the Seventh Session of the Codex Committee on

2.4  rinciples governing the toxicological evaluation of compounds

2.5 Food additive specifications

2.5.1 Requirements for submission of analytical methods

The Committee, while encouraging the use of analytical methods published

2.5.2 Analytical method for the determination of residual solvents by

2.5.3 Analytical method for the determination of carbon number at 5%

2.6 GEMS/Food consumption data

3.1 Safety evaluations

Chemical and technical considerations

Assessment of dietary exposure

3.1.2 Glucoamylase from Trichoderma reesei expressed in Trichoderma

Chemical and technical considerations

Assessment of potential allergenicity

Assessment of dietary exposure

3.1.3 Glycerol ester of gum rosin

Chemical and technical considerations

3.1.4 Glycerol ester of tall oil rosin

3.1.5 Glycerol ester of wood rosin

Chemical and technical considerations

Chemical and technical considerations

Assessment of dietary exposure

3.2 Revision of specifications

3.2.1 Annatto extracts (solvent-extracted bixin and solvent-extracted

3.2.2 Benzoe tonkinensis

The specifications of Benzoe tonkinensis prepared at the seventy-fourth meet-

3.2.3 Food additives containing aluminium and/or silicon

The Committee at its seventy-sixth meeting (Annex 1, reference 211)

3.2.4 Food additives containing phosphates: Analytical methods for the

3.2.5 Mineral oil (medium viscosity)

The Committee at its seventy-sixth meeting (Annex 1, reference 211) drafted

3.2.6 Modified starches

The specifications for modified starches cover 16 different modified starches,

3.2.7 Paprika extract

The sixty-ninth meeting of the Committee (Annex 1, reference 190) prepared

3.2.8 3-Phytase from Aspergillus niger expressed in Aspergillus niger

The Committee at its seventy-sixth meeting (Annex 1, reference 211) had

3.2.9 Potassium aluminium silicate

3.2.10 Potassium aluminium silicate–based pearlescent pigments

The Committee, at its seventy-fourth meeting (Annex 1, reference 205), pre-

1.Food additives - analysis. 2.Food additives - toxicity. 3.Food contaminants - analysis.

Annex 1 Reports and other documents resulting from previous

Annex 2 Toxicological and dietary exposure information and

2.2 eport from the Forty-fifth Session of the Codex Committee on

2.3 eport from the Seventh Session of the Codex Committee on

2.4 rinciples governing the toxicological evaluation of compounds

2.5.2 Analytical method for the determination of residual solvents by

2.5.3 Analytical method for the determination of carbon number at 5%

3.1.2 Glucoamylase from Trichoderma reesei expressed in Trichoderma

3.2.1 Annatto extracts (solvent-extracted bixin and solvent-extracted

3.2.4 Food additives containing phosphates: Analytical methods for the

4.1 admium: Assessment of exposure from cocoa and cocoa

nalytical method for the determination of carbon number at

nalytical method for the determination of residual solvents by

hosphates: Analytical methods for the determination of

nnatto extracts (solvent-extracted bixin and solvent-extracted

succinate was revised.