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    WORLD HEALTH ORGANIZATION

    WHO Food Additives Series 1972, No. 1




    TOXICOLOGICAL EVALUATION OF SOME 
    ENZYMES, MODIFIED STARCHES AND 
    CERTAIN OTHER SUBSTANCES




    The evaluations contained in this publication were prepared by the
    Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
    16-24 June 19711





    World Health Organization

    Geneva

    1972





                   

    1 Fifteenth Report of the Joint FAO/WHO Expert Committee on Food
    Additives, Wld Hlth Org. techn. Rep. Ser., 1972, No. 488; FAO
    Nutrition Meetings Report Series, 1972, No. 50.

    The monographs contained in the present volume are also issued by the
    Food and Agriculture Organization of the United Nations, Rome, as FAO
    Nutrition Meetings Report Series, No. 50A

    (c) FAO and WHO 1972

    CONTENTS

    INTRODUCTION

    ALLOCATION OF ACCEPTABLE DAILY INTAKES

    TOXICOLOGICAL EVALUATION: ENZYMES

    TOXICOLOGICAL EVALUATION: MODIFIED STARCHES

    TOXICOLOGICAL EVALUATION: MISCELLANEOUS FOOD ADDITIVES

    THE MONOGRAPHS

    ENZYMES

    Mixed Microbial Carbohydrase and Protease
    Microbial Carbohydrase
    Microbial Glucose Oxidase
    Microbial Rennet
    Microbial Rennet
    Microbial Rennet

    MODIFIED STARCHES

    Oxidized Starches
    Starch Acetate
    Hydroxypropyl Starch
    Hydroxypropyl Distarch Glycerol
    Distarch Phosphate
    Hydroxypropyl Distarch Phosphate
    Phosphated Distarch Phosphate
    Acetylated Distarch Phosphate
    Distarch Glycerol
    Acetylated Distarch Glycerol
    Acetylated Distarch Adipate

    MISCELLANEOUS FOOD ADDITIVES

    Caramel Colours (Ammonia process)
    Esters of Glycerol and Thermally Oxidized Soy Bean Fatty Acids
    Hexamethylenetetramine
    Microcrystalline Cellulose
    Propylene Glycol Alginate
    Stearoyl Lactylic Acid, Calcium and Sodium Salts
    Tin and Stannous Chloride

    ANNEX 1. Reports and other documents resulting from previous meetings
    of the Joint FAO/WHO Expert Committee on Food Additives

    ANNEX 2. Membership of the Fifteenth Joint FAO/WHO Expert Committee on
    Food Additives
    

    INTRODUCTION

    The monographs contained in this document are the result of the
    deliberations of the Joint FAO/WHO Expert committee on Food Additives
    which met in Rome from 16 to 24 June 1971. Since the report1 of this
    meeting also contains comments on groups of as well as individual
    substances considered, these monographs should be read in conjunction
    with the report.

    Any new information and comments relating to the biological data and
    their evaluation should be addressed to: Food Additives unit, World
    Health Organization. Avenue Appia, Geneva, Switzerland.

    Specifications on the food additives considered have been prepared,
    and these will be issued in a separate document entitled
    "Specifications for the Identity and Purity of Some Enzymes and
    Certain Other Substances".2  Comments relating to specifications
    should be addressed to: Food Policy and Food Science Service,
    Nutrition Division, FAO, Rome, Italy.

                   

    1 Fifteenth Report of the Joint FAO/WHO Expert Committee on Food
    Additives, FAO Nutrition Meetings Report Series; Wld Hlth Org. techn.
    Rep. Ser., 1972, No. 488.

    2 FAO Nutrition Meetings Report Series, WHO/Food Add./71 (In
    preparation).

    ALLOCATION OF ACCEPTABLE DAILY INTAKES

    It is generally recognized that all chemicals are toxic to animals and
    man if large enough amounts are administered.  Even those so-called
    innocuous substances, when given in excessive doses, may induce
    untoward effects as a result of various non-specific actions, e.g.
    physical obstructions of the G.I. tract, alteration of osmotic
    pressure and nutritional imbalance.  A limit on the daily intake of a
    substance is therefore essential for the protection of the health of
    the consumer.  The Committee therefore followed previous practice,
    where appropriate, in allocating acceptable daily intakes (ADI) to the
    food additives considered.

    An unconditional ADI was allocated only to those substances for
    which the biological data available included either the results of
    adequate short-term and long-term toxicological investigations or
    information on the biochemistry and metabolic fate of the compound or
    both.

    A conditional ADI was allocated for specific purposes arising from
    special dietary requirements.  (None was allocated at this meeting.)

    A temporary ADI was allocated when the available data were not fully
    adequate to establish the safety of the substance and it was
    considered necessary that the additional evidence be provided within a
    stated period of time.  If the further data requested do not become
    available within the stated period, it is possible that the temporary
    ADI will be withdrawn by a future committee.

    Whereas all the ADIs are subject to periodic review, especially when
    additional data are available, reconsideration of temporary ADIs is
    obligatory after the date specified.  The decision arrived at will
    depend on the information available at the time of review.  It is
    certainly not intended that the temporary status of the ADI for a food
    additive will continue to be maintained indefinitely.

    "Not limited". The Committee took cognizance of the fact that the
    level of use of certain food additives is limited by good
    manufacturing practice and that they are food components or possess a
    very low toxicity.  For these additives the Committee established no
    limit except as based on good manufacturing practice.

    For those additives for which the available information was grossly
    inadequate to establish safety, no ADI was allocated.

        TOXICOLOGICAL EVALUATION: ENZYMES

                                                                                    

                                        Evaluation1     Monographs    Specifications
                                                                                    

    I.   Animal-derived enzyme preparations

    1.   Rennet                         Not limited       No            Yes
    2,   Rennet, animal                 Not limited       No            Yes
    3.   Pepsin                         Not limited       No            Yes
    4.   Trypsin                        Not limited       No            Yes
    5.   Lipase, animal                 Not limited       No            Yes
    6.   Catalase (bovine liver)        Not limited       No            Yes

    II.  Plant-derived enzyme preparations

    1.   Bromelain                      Not limited       No            Yes
    2.   Ficin                          Decision          No            Yes2
                                        postponed
    3.   Malt-carbohydrases             Not limited       No            Yes
    4.   Papain                         Not limited       No            Yes

    III. Microbially-derived enzyme preparations

    1.   Aspergillus niger var.

         (a)  Carbohydrase              Not limited3      Yes           Yes2
         (b)  Glucose oxidase           Not limited3      Yes           Yes2

    2.   Aspergillus oryzae var.

         (a)  Carbohydrases             Decision          No            Yes2
                                        postponed
         (b)  Protease                  Decision          No            Yes2
                                        postponed
         (c)  Lipase                    Decision          No            Yes2
                                        postponed
                                                                                    

    TOXICOLOGICAL EVALUATION: ENZYMES (continued)

                                                                                    

                                        Evaluation1     Monographs    Specifications
                                                                                    

    3.   Bacillus cereus

         Microbial rennet               Decision          No            Yes2
                                        postponed

    4.   Bacillus subtilis var.

         Carbohydrases and              Not limited       Yes           Yes
         proteases mixed

    5.   Endothia parasitica

         Microbial rennet               Not limited3      Yes           Yes2

    6.   Mucor miehei

         Microbial rennet               Not limited3      Yes           Yes2

    7.   Mucor pusillus

         Microbial rennet               Not limited3      Yes           Yes2

    8.   Micrococcus lysodeikticus

         Catalase                       Decision          No            Yes2
                                        postponed

    9.   Rhizopus oryzae

         Carbohydrase                   Not limited       No            Yes
                                                                                    

    TOXICOLOGICAL EVALUATION: ENZYMES (continued)

                                                                                    

                                        Evaluation1     Monographs    Specifications
                                                                                    


    10.  Saccharomyces sp.

         Carbohydrase                   Not limited       No            Yes

    11.  Miscellaneous

         Irpex lacteus                  Decision          No            No
         (microbial rennet)             postponed

         Penicillium amagasakiense      Decision          No            No
         (microbial glucose oxidase)    postponed

         Arthrobacter                   Decision          No            No
         (microbial carbohydrase)       postponed
                                                                                    

    1 Not limited, except by good manufacturing practice.
    2 Tentative specifications.
    3 Temporary.

    TOXICOLOGICAL EVALUATION: MODIFIED STARCHES1

                                                                                    

                                        Evaluation2     Monograph     Specification
                                                                                    

    Oxidized starches                   Not limited     Yes           Yes

    Starch acetate                      Not limited     Yes           Yes

    Hydroxypropyl starch                Not limited     Yes           Yes

    Hydroxypropyl distarch
         glycerol                       Not limited     Yes           Yes

    Distarch phosphate                  Not limited     Yes           Yes

    Phosphated distarch phosphate       Not limited     Yes           Yes

    Acetylated distarch phosphate       Not limited     Yes           Yes

    Distarch glycerol                   Not limited     Yes           Yes

    Acetylated distarch glycerol        Not limited     Yes           Yes

    Acetylated distarch adipate         Not limited     Yes           Yes

    Hydroxypropyl distarch              Not             Yes           Yes3
         phosphate                      acceptable
                                                                                    

    1 For results of evaluation on other modified starches see Annex 4, Thirteenth 
      Report, Wld Hlth Org. techn. Rep. Ser., No. 445.
    2 Use limited by good manufacturing practice; temporary acceptance.
    3 Tentative specification.

    TOXICOLOGICAL EVALUATION: CERTAIN OTHER SUBSTANCES

                                                                                    
                                        Acceptable
                                        daily intake    Monograph     Specifications
         Substances                     for man1
                                        (mg/kg
                                        body weight)
                                                                                    
    Caramel colours
    (a)  made by the ammonia or
         ammonium sulfate process         0-1002,3        Yes           Yes6
    (b)  made by other processes        Not limited4      No            Yes

    Choline salts                       Not limited4      No            No

    Esters of glycerol and thermally
    oxidized soy bean fatty acid          0-752           Yes           Yes6

    Thermally oxidized soy bean oil       No ADI          No            Yes6

    Hexamethylenetetramine                0-52            Yes           Yes

    Microcrystalline cellulose          Not limited4      Yes           Yes

    Propylene glycol alginate             0-25            Yes           Yes

    Stearoyl lactylate, calcium and
         sodium salts                     0-20            Yes           Yes

    Tin and stannous chloride           No ADI5           Yes           Yes
                                                                                    

    1 Unconditional unless otherwise stated. 
    2 Temporary ADI.
    3 The level of 4-methylimidazole should not exceed 200 mg/kg based on a product
      having a colour intensity of 20 000 EBC (European Brewing Convention) Units.
    4 Use limited by good manufacturing practice.
    5 See the Monograph.
    6 Tentative specifications.




























    See Also:
       Toxicological Abbreviations