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    SUCROSE ESTERS OF FATTY ACIDS AND SUCROGLYCERIDES

    First draft prepared by
    Ms Elizabeth Vavasour
    Toxicological Evaluation Division, Bureau of Chemical Safety
    Food Directorate, Health Protection Branch, Health Canada
    Ottawa, Ontario, Canada

    Explanation
    Biological data
         Biochemical aspects
         Absorption, distribution, and excretion
    Toxicological studies
         Short-term toxicity studies
         Long-term toxicity/carcinogenicity studies
         Special studies on gastrointestinal effects
         Observations in humans
    Comments
    Evaluation
    References

    1.  EXPLANATION

         Sucrose esters of fatty acids and sucroglycerides were previously
    considered by the Committee at its thirteenth, seventeenth, twentieth,
    twenty-fourth, thirty-fifth and thirty-ninth meetings (Annex 1,
    references 19, 32, 41, 53, 88 and 101). At the thirty-ninth meeting,
    the Committee established a group ADI of 0-16 mg/kg bw for sucrose
    esters contained in sucrose esters of fatty acids and sucroglycerides,
    based on a NOEL for decreased body-weight gain of 50 g/kg in the diet,
    equal to 1.6 g/kg bw/day, observed in a long-term carcinogenicity
    study in rats with a palm oil sucroglyceride. The NOEL was corrected
    for the content of mono- and diglycerides in the test material since
    these substances were considered to be normal constituents of the
    human diet. Because sucrose esters are hydrolysed in the gut to normal
    dietary constituents prior to absorption, the Committee used a safety
    factor of 50 in deriving the ADI.

         Since the last evaluation, new studies have become available and
    are discussed in the following monograph addendum.

    2.  BIOLOGICAL DATA

    2.1  Biochemical aspects

    2.1.1  Absorption, distribution, and excretion

    2.1.1.1  Rats

         Four studies were conducted in male and female F344 rats. In the
    first study, test material with the same composition as that employed
    in the 13-week (section 2.2.2.1) and long-term feeding studies
    (section 2.2.3) was used1. S-570 was administered as a single oral
    dose of 50, 100 or 200 mg/kg bw (males) or 100 mg/kg bw (females), or
    as a single i.v. injection of 1 mg/kg bw (males only). Blood samples
    were collected from the vena cava of 4 animals/group at varying
    intervals for 24 h after dosing and sucrose monostearate (SMS)
    concentrations in the plasma were determined using GC-MS. Following
    oral administration, peak plasma concentrations of SMS (Cmax in the
    males were reached at 1, 2 and 2 h (Tmax) at doses of 50, 100 and
    200 mg/kg bw, respectively. The half-lives for elimination of SMS from
    the plasma were 4.1, 2.4 and 3.1 h for these same groups, suggesting
    that SMS was eliminated from the plasma within 24 h of ingestion. In
    females receiving 100 mg/kg bw, values for Tmax and T´ were
    consistent with those observed in their male counterparts, while the
    values for Cmax and AUC0-infinity were twice those in the males.
    The bioavailability of SMS was very low, 0.26 - 0.33%.

         Diets containing 1% or 5% S-570 were fed to groups of 4 male rats
     ad libitum for periods of 1, 2 or 4 weeks and for 4 weeks followed
    by a 3-day, 1-week or 2-week recovery period. At the end of the study
    period, blood samples were taken from the vena cava, livers, kidneys,
    hearts, lungs, and spleens, and perirenal fat was collected for
    determination of sucrose monoester content. The mean intake of S-570
    after 1, 2, 3 and 4 weeks of treatment was 720, 690, 640 and 590 mg/kg
    bw/day, respectively, from the 1% diet, and 3670, 3520, 3370 and
    3030 mg/kg bw/day from the 5% diet. Following dietary administration
    of S-570 at the 1 % dietary level, SMP (sucrose monopalmitate)
    concentrations in the plasma and tissues were close to or below the
    limits of detection (0.01 - 0.05 µg/g tissue). At the 5% dietary
    level, similar amounts of SMP were detected at the sampling points in
    most of the tissues, with the highest concentrations in the liver
    (0.3-0.4 µg/g tissue), followed by kidney, lung, spleen, heart and
    plasma. SMS was detected in tissues and plasma at both dose levels

              

    1   S-570: 28% monoesters, 34% diesters, 21% triesters and 10% tetra-
        and higher esters, with a fatty acid composition of 70:30 stearic
        to palmitic acid.

    with concentrations following the same order as for SMP (liver
    highest, followed by kidney, lung, spleen) and proportional to the
    dose. At the 5% dose, the tissue concentrations for the liver, spleen
    and lung increased with duration of treatment. No sucrose monoesters
    were detected in any of the tissues 3 days after treatment was
    discontinued except for the heart in the high-dose groups, in which
    concentrations of monoesters were below the limits of detection 7 days
    after discontinuation of treatment. The retention ratios (tissue
    content in relation to final daily dose) were higher for SMS than for
    SMP, by about 4-5 fold in the liver and kidney, and < 2-3-fold for
    the heart, spleen and lung.

         Plasma and liver tissue were collected from 5 rats/sex/group at
    the termination of the long-term study described in section 2.2.3. In
    this study, dietary levels of 1%, 3% or 5% were fed to rats for 2
    years. The SMS content of tissues and plasma was greater than that of
    SMP. SMP content of the plasma was below or close to the detection
    limit, while SMS content of the plasma (0.02-0.15 µg/ml) and SMS and
    SMP content of the liver (0.1-0.6 µg/ml and 0.01-0.09 µg/ml,
    respectively) were proportional to the dose. The retention ratios for
    SMS and SMP in the liver and plasma were comparable to those in the
    1-4 week study described above, indicating that no accumulation of
    monoesters occurred in these tissues.

         Male rats received by gavage 100 mg/kg bw of 14C-SMS or
    14C-SDS (sucrose distearate), labelled on stearic acid, by gavage.
    Radioactivity in the blood reached a peak (12.0 µg eq. SMS/ml and
    8.4 µg eq. SDS/ml) at 3 h after dosing. Elimination of radioactivity
    from the blood was biphasic, with half-lives of approximately 35 h and
    90 h for each of the phases. After 168 h (1 week), radioactivity in
    the blood had decreased to 14% of the peak levels. The cumulative
    excretion at 24 h of radioactivity associated with SMS was 1.4%, 31%
    and 29% in the urine, faeces and expired air, respectively. By 168 h,
    2%, 35% and 36% of radioactivity had been excreted by these routes and
    18% was retained in the carcass. With SDS, cumulative excretion of
    radioactivity in the urine, faeces and expired air at 24 h was 0.7%,
    63% and 13%, respectively. After 168 h, cumulative excretion by these
    routes was 1%, 67% and 17%, with 9% remaining in the carcass
    (Mitsubishi Institute, 1994a).

         In studies designed to determine the contribution of biliary
    excretion to elimination of mono- and diesters, bile-duct cannulated
    male rats received an i.p. dose of 39 mg/kg bw of 14C-SMS or
    14C-SDS. Biliary excretion accounted for less than 0.1% of the
    administered radioactivity 8 h after dosing. After 48 h, cumulative
    excretion of radioactivity from SMS was 0.1%, 0.5% and 9% and from SDS
    was <0.1%, 0.1% and 16% in the bile, urine and faeces, respectively.
    Since biliary excretion for both compounds was negligible, GI tract
    absorption of radioactivity was from SMS 56%, and from SDS 27% (based
    on combined radioactivity excreted in the urine and expired air, and
    retained in the carcass). The distribution of radioactivity to tissues

    24 h after gavage administration of 14C-SMS or 14C-SDS was highest
    in the liver, followed by skin, muscle, white fat, blood and kidney
    (SMS only). After 168 h, the order of distribution of radioactivity
    had changed to white fat, muscle, skin, liver, kidney (SMS only) and
    pancreas. Urine, faeces, plasma and various tissue samples were
    assayed for SMS, SDS and metabolites. After SMS administration, very
    little unchanged SMS was excreted in the urine or faeces (representing
    1.4% and 2.0% of radioactivity excreted at 24 h by these routes,
    respectively). The major faecal metabolite (87% of faecal
    radioactivity) was identified as stearic acid. The highest
    concentrations of unchanged SMS were detected at 2 and 4 h after
    dosing in the lung and liver, and SMS was detected at lower levels in
    a number of other organs representing less than 0.1% of the total
    administered radioactivity. After 24 h, SMS was not detected in any of
    the tissues or plasma. When SDS was administered, a very small amount
    of unchanged SDS was detected in the urine (2.2% of urinary
    radioactivity, 0.01% of the total dose). Both SDS and SMS were present
    in the faeces. SDS and stearic acid were the major radioactive
    components in the faeces (39% and 51% of faecal radioactivity,
    respectively). SMS was a minor component (4.3% of faecal
    radioactivity). Neither SDS nor SMS were detected in the plasma or
    tissues at 2, 4 or 24 h after administration. The identity of major
    plasma and tissue metabolites were not identified. The presence of
    unchanged SMS or SDS in urine was attributed to contamination from
    faeces during collection (Mitsubishi Institute, 1994a).

    2.1.1.2  Dogs

         Three male beagle dogs were given S-11701 in a single gelatin
    capsule at doses of 50, 250, and 1250 mg/kg bw in that order,
    separated by a washout period of 7 days between the low and mid doses
    and a period of 12 days between the mid and high doses. The amounts of
    SMP in these doses were 9, 44 and 221 mg/kg bw, respectively; the
    amounts of SMS were 21, 103 and 514 mg/kg bw, respectively. Blood
    samples were obtained over a 48-hour period following administration
    of each dose and SMP and SMS concentrations in the plasma were
    determined using GC-MS. Both SMP and SMS were detected in the plasma
    at concentrations which were proportional to the dose administered
    (0.06-0.60 µg/ml and 0.12-1.14 µg/ml, respectively). Peak plasma
    concentrations occurred at 3.3-4.7 h for SMP and 3.3-7.3 h for SMS,
    and increased with increasing dose. SMP was eliminated from plasma
    with half-lives of 2.5 and 5.6 h for the mid and high doses; for SMS
    the elimination half-lives from the plasma were 7.2 and 7.3 h for the
    mid and high doses (Mitsubishi Institute, 1994a).

              

    1    sucrose esters of stearic and palmitic acids, with 57%
         monoester, 28 diester, 10% triester and 1% tetra and higher
         esters, and a ratio of 70:30, stearate to palmitate.

    2.1.1.3  Humans

         A clinical study was conducted with a total of 11 volunteers in 3
    single dose segments (3, 3 and 6 subjects) and one multiple dose
    segment (5 subjects). The subjects were healthy male adults, 22 to 28
    years old and 51.5 to 70 kg weight. In the single dose experiments,
    the subjects drank 1, 2 or 3 g S-1170 in 200 ml orange juice once a
    day; in the multiple dose experiment, the subjects drank 1 g S-1170 in
    200ml orange juice, twice daily (2 g/day) for 5 days. Doses of 1, 2 or
    3 g were equivalent to 17, 33 or 50 mg/kg bw for a 60-kg person. Blood
    samples were collected for determination of SMP and SMS content. Urine
    and faeces were collected for determination of mono-, di-, and
    triesters. Both SMS and SMP were detected in the plasma at 2 and 6 h
    at concentrations of 0.01-0.04 µg/ml which was close to the limit of
    detection (0.01 µg/ml). Concentrations of SMS close to the detection
    limit were detectable 24 h after ingestion of S-1170 following dosing
    with 2 g S-1170, and in the multiple dose study. SMS was detected at
    slightly higher levels than SMP, reflecting the higher content of the
    former in S-1170.

         No unchanged mono-, di- or triesters were detected in urine
    following single or multiple doses. Faecal excretion of sucrose esters
    was 22%, 25% and 31% of a single dose of 1, 2 or 3 g S-1170 after
    48 h. After the multiple dose regimen, 17% of the total dose was
    excreted in the faeces as sucrose esters. This suggested a hydrolysis
    rate of 70-80%. The composition of individual esters excreted in the
    faeces indicated that greater hydrolysis of monoesters had occurred
    compared with higher esters and that SMP was hydrolysed to a greater
    degree than SMS.

         Cultures of human intestinal flora were incubated at 37°C for 5 h
    with S- 1170 added to give final concentrations of 0.05%, 0.25% or
    0.5%. Analysis of the total sucrose ester content following incubation
    indicated that 52%, 69% and 68%, respectively, of the sucrose esters
    bad been hydrolysed to sucrose and free fatty acids.

         SMP, SMS and S-1170 were incubated with artificial gastric juice
    (pH 1.2) at a concentration of 0.4, 0.4 and 1.3 mg/ml, respectively.
    Determination of mono-, di- and triesters were carried out with
    HPLC-MS after incubation periods of up to 5 h at 37°C. After 5 h
    incubation, 82% of SMP and 85% of SMS remained. When S-1170 was
    tested, the proportion of unhydrolysed SMP, SMS, diesters and
    triesters were 83%, 93%, 99%, and 94%, respectively (Mitsubishi
    Institute, 1994a).

         In a supplementary study, human volunteers received S-1170 which
    had been baked into bread. A number of single and multiple dosing
    regimes were employed which involved doses ranging from 1-2 g (see
    section 2.3). SMS and SMP were detected in plasma after single dose
    administration from 2 h after dosing, and reached a peak at 6 h (SMP

    0.02-0.04 µg/ml; SMS 0.07-0.11 µg/ml). By 24 h after administration,
    the monoesters were not detectable except for a few samples at the
    detection limit (0.01 µg/ml) in the subjects receiving a dose of 2 g.
    Following multiple exposures (1.5 g, 3 times/day, 1 or 7 days),
    monoester concentrations in the plasma gradually increased, reaching a
    plateau by 3 days (SMP 0.084-0.14 µg/ml; SMS 0.19-0.33 µg/ml). SMP was
    not detectable 24 h after the last dose, and SMS was at the limit of
    detection at 24 h and not detectable by 24 h (Mitsubishi Institute,
    1994b).

    2.2  Toxicological studies

    2.2.2  Short-term toxicity studies

    2.2.2.1  Rats

         Groups of 20 male and female F344/DuCrj rats were fed diets
    containing 0, 1%, 3% or 5% S-570 for 13 weeks (a mixture of sucrose
    esters of stearic and palmitic acids and traces of oleic acid,
    containing no mono- and di- glycerides). These dietary concentrations
    were equal to 0, 636, 1900 or 3240 mg/kg bw/day in males and 0, 666,
    1950 or 3430 mg/kg bw/day in females. The animals were observed daily.
    Physical examinations, and determination of body weights and food
    consumption were performed weekly. A standard set of haematological
    and clinical chemistry parameters were measured in blood samples taken
    from all animals at termination. Urinalysis parameters were measured
    prior to termination in 10 rats/sex/group. Ophthalmological
    examinations were conducted on 10 rats/sex/group prior to initiation
    of treatment and on 10 rats/sex from the control and high-dose groups
    before the end of treatment. Gross necropsy was performed on all
    animals at termination, and organ weights for liver, kidneys,
    adrenals, testes, ovaries, brain, heart, lungs and spleen were taken.
    Histopathological examinations of 44 tissues and organs and gross
    lesions in all control and high-dose animals and in female tissues
    showing macroscopic changes were carried out.

         Administration of the test material had no consistent effect on
    body-weight gain, food consumption or food efficiency. There were
    no toxicologically significant effects of treatment on the
    haematological, clinical chemistry or urinalysis parameters. No
    abnormalities were reported as a result of ophthalmological
    examinations. No macroscopic, organ weight or histo-pathological
    changes were observed which could be attributed to treatment
    (Mitsubishi-Kasei Institute 1991).

    2.2.3  Long-term toxicity/carcinogenicity studies

    2.2.3.1  Rats

         Ryoto Sugar Ester S-570 was administered in the diet to male and
    female F344/DuCrj rats at concentrations of 0, 1%, 3% or 5%, equal to
    doses of 0, 394, 1160 or 1970 mg/kg bw/day in male rats and 0, 480,
    1440 or 2440 mg/kg bw/day in female rats. The study was divided into
    two parts, a 2-year carcinogenicity assay employing 50 rats/sex/group
    and a 52-week chronic toxicity assay in which satellite groups of 14
    rats/sex/group were used. Each animal was observed daily for 74 weeks
    and twice daily thereafter. Detailed physical examinations were
    conducted weekly. Body weight and food consumption data were collected
    weekly for the first 13 weeks and at least once every 4 weeks for the
    remainder of the study. Blood samples were collected from all animals
    in the chronic toxicity phase of the study for determination of a
    standard set of haematological and clinical chemistry parameters
    before the start of the study (clinical chemistry only), every 3
    months during the study, and at termination. For rats in the
    carcinogenicity portion of the study, blood samples were collected
    every 6 months and at termination for determination of haematological
    parameters only. Ophthalmological examinations were conducted on all
    male and female rats in the chronic toxicity phase before test
    substance administration, and on the control and high-dose male and
    female rats every 3 months during the study up to one week before
    termination. Gross necropsy was performed on all animals dying on test
    or at scheduled sacrifices at 52 weeks (chronic toxicity phase) and
    104 weeks (carcinogenicity assay). At the scheduled sacrifices, liver,
    kidneys, adrenals, testes, ovaries, brain, heart, lungs and spleen
    weights were recorded. Histopathological examinations were made of 48
    tissues and organs (including liver and tissues of the GI tract), of
    gross lesions in rats from the control and high-dose groups, and of
    all animals dying on test. The liver, lungs, kidneys and tissues
    showing macroscopic changes were examined in animals from the low- and
    mid-dose groups.

         Treatment with the test material had no effect on survival. No
    deaths occurred in the chronic toxicity phase of the experiment and
    the survival rates at 104 weeks were similar (66-76%) between treated
    groups and controls. Body weights in the male rats receiving 5% test
    material were lower than in controls at various points during the
    first year of the experiment, but the decrements were only 2-3%. In
    the low-dose female rats, body weights were occasionally significantly
    higher compared with controls. This occurred mostly toward the end of
    the study and the increases were in the order of 6-7% of controls.
    During the first week of administration, food consumption was
    significantly reduced in both males and females receiving the high
    dose (6% and 4% of controls, respectively). Occasional reductions in
    food consumption of the high-dose male rats were noted throughout the
    rest of the study, but these were small (3-6%). Food efficiency was

    not affected by inclusion of test material in the diet of either the
    male or female rats. Statistically significant increases in the mean
    corpuscular volume affecting mainly the high-dose males and females
    were noted at many of the sampling points, as well as sporadic
    significant decreases toward the end of the study in red blood cell
    count, haemoglobin concentration, MCHC and platelet count. Although
    statistically significant, the actual differences from controls
    were small and not toxicologically significant. No effects of
    treatment were evident from the clinical chemistry parameters or
    ophthalmological examinations. Absolute and relative spleen weights
    were significantly greater in the mid- and high-dose males and the
    mid-dose females (non-significant in high-dose females) compared with
    controls at the end of 104 weeks. The spleen weight changes and
    changes in haematological parameters were likely due to the incidence
    of large granular lymphocyte leukaemia, which was a commonly occurring
    neoplasm in all groups and was slightly increased without statistical
    significance at the two top doses in both sexes (7/50, 9/50, 11/50 and
    12/50 in order of ascending dose in males; 10/50, 7/50 14/50 and 13/50
    in females). At the time of organ weight determination, there were
    2/36, 4/34, 6/35 and 5/34 male rats affected and 4/38, 1/35, 4/35 and
    2/33 females affected. When leukaemic animals were removed from spleen
    weight determinations, no treatment-related difference was apparent.
    Consistent with the higher incidence of leukaemia in the high-dose
    group, the incidence of associated non-neoplastic findings,
    extramedullary haematopoiesis in the spleen and haematopoietic
    hyperplasia in the bone marrow was increased in the high-dose groups.
    Historical control incidences of LGL leukaemia as high as 24% in males
    and 25% in female F344 rats have been noted in the laboratory
    conducting the study. Consequently, the LGL leukaemia was not likely
    an effect of treatment, nor were the observed effects on the spleen,
    bone marrow and haematological parameters. There were no increases in
    the incidence of other neoplastic or non-neoplastic lesions which
    could be attributed to treatment. The NOEL was 5% of the diet, equal
    to 1970 mg/kg bw/day (Mitsubishi Institute, 1994c).

    2.2.4  Special studies on gastrointestinal effects

         Sucrose monostearate (SMS), a component of sucrose esters, was
    assayed for its effect on intestinal contractility in an isolated
    rabbit ileum preparation. A concentration of 0.25 mg/ml, but not
    0.025 mg/ml transiently elevated muscle tonus in the relaxation phase
    of contraction.

         The intestinal transport rate for Sugar Ester S-1170 was
    determined in groups of 8 fasted mice. A suspension of the test
    material in 0.5% gum arabic solution was administered by gavage at
    doses of 0, 2500 or 5000 mg/kg bw. One hour later, a 5% suspension of

    charcoal in 5% gum arabic solution was administered by gavage at a
    volume of 0.2 ml/animal. After 30 minutes, the GI tract was measured
    and the distance travelled by the charcoal from the pylorus was
    measured as a percentage of the total. Both doses of sucrose ester
    significantly reduced the transit time of the GI tract.

         SMS was administered intravenously to dogs. Three dogs each
    received 1, 2 and 4 mg/kg bw of SMS sequentially followed by an
    observation period of one week. Loose faeces and/or diarrhoea were not
    observed as an effect of treatment. The investigators concluded that
    the laxative action of sucrose esters was not induced by absorbed
    ester (Mitsubishi Institute 1994d).

    2.3  Observations in humans

         In a supplement to the pharmacokinetic study in which S-1170 was
    administered to human volunteers in orange juice (Mitsubishi
    Institute, 1994a), sucrose ester S-1170 was administered in bread and
    the clinical observations from both studies were compared. The
    subjects were healthy males, 20-29 years of age and weighing 60 ±
    10 kg. In this study, groups of five subjects ate bread rolls
    containing 1, 1.5 or 2 g of S-1170 at a single meal (single dose),
    bread rolls containing 1.5 g S-1170 at 3 meals per day (4.5 g/day) for
    1 or 7 days (3 or 21 doses), and bread rolls containing 1 g S-1170 at
    2 or 3 meals (2 or 3 g/day) a day for 5 days (10 or 15 doses).
    Subjective symptoms were recorded throughout the study, physical
    examinations were conducted and haematology, clinical chemistry and
    urinalysis parameters measured.

         Following a single dose of sucrose esters in orange juice (see
    section 2.1.1.3 for the protocol), soft stools or diarrhoea were
    observed in 4/6 and 3/3 subjects receiving 2 or 3 g test substance,
    but not in the subjects receiving 1 g either as a single dose or twice
    daily for 5 days. The severity as well as the incidence increased
    with dose. When the test substance was administered in bread,
    treatment-related observation of soft stools or diarrhoea was noted in
    1/5 subjects receiving a single dose of 1.5 g and in 3/5 subjects
    receiving 2 g. In the multiple dose studies, treatment-related
    increases in laxation were observed in 4/5 subjects receiving 1.5 g,
    3 times daily for 7 days (1-5 events), in 2/5 subjects receiving 1 g,
    3 times daily for 5 days, and in 1/5 subjects receiving 1 g 2 times
    daily for 5 days. Clinical symptoms other than laxation were bloated
    feeling, borborygmus, abdominal pain, flatus, suprapubic discomfort
    and nausea. These symptoms were noted 1 to 10 h after ingestion of
    S-1170 and tended to subside by 24 h. There were no treatment-related
    changes in the results of the physical examinations or in the
    haematology, clinical chemistry and urinalysis parameters (Mitsubishi
    Institute, 1994b).

    3.  COMMENTS

         Since the last evaluation, new long-term toxicity/carcinogenicity
    and pharmacokinetic studies have become available in which the test
    material was composed entirely of sucrose esters of stearic and
    palmitic acids (70:30), with traces of oleic acid esters. In contrast
    to the sucrose esters previously tested, which consisted mainly of
    mono- and diesters, together with traces of triesters, the test
    material administered to rats in these studies contained 28%
    monoesters, 34% diesters, 21% triesters and 10% higher esters, while
    that administered to humans and dogs contained 57% monoesters, 28%
    diesters, 10% triesters and 1% higher esters. Neither of these sucrose
    ester formulations contained any mono- or diglycerides.

         No adverse effects of treatment were demonstrated in the
    long-term toxicity/carcinogenicity study conducted in rats at dose
    levels up to 50 g/kg in the diet, equal to 1970 mg/kg bw/day.

         A series of studies on the disposition of two sucrose ester
    formulation in rats, dogs and humans demonstrated that small amounts
    of the monoesters were absorbed in all three species. On the basis of
    tissue residue and excretion studies in rats, it appears unlikely that
    diesters were absorbed. The small amounts of monoesters absorbed
    intact were completely metabolized and either excreted as carbon
    dioxide or integrated into body components. In humans, 20-30% of a
    dose of sucrose esters was retrieved intact from the faeces following
    a single or multiple dosing regimen, suggesting that 70-80% of the
    dose was hydrolyzed to sucrose and component fatty acids.

         The results of human tolerance studies conducted with sucrose
    esters administered in orange juice or with bread were also available.
    Although there were significant deficiencies in this study, most
    notably in the size of the study groups and the lack of any controls,
    the results, indicating laxation and related abdominal symptoms with
    treatment at single doses of 1.5-3 g or divided doses of 3-4.5 g per
    day for 5-7 days, were of concern. A divided dose of 2 g per day for
    5 days, equal to 35 mg/kg bw/day, produced no effect when administered
    in orange juice, and only a slight effect in 1/5 subjects when
    administered with bread. Only the highest dose tested, namely 4.5 g
    per day for 7 days, equal to 70 mg/kg bw/day, resulted in multiple
    occurrences of GI symptoms (soft stools, diarrhoea, flatulence,
    borborygmus and bloated sensation) in the subjects. The results in
    humans were of interest since doses of 2000 mg/kg bw/day did not
    induce GI disturbances in rats.

    4.  EVALUATION

         In view of the reservations about the tolerance study in humans,
    and a demonstrated NOEL in the rat of approximately 2000 mg/kg bw/day
    for sucrose esters of fatty acids, the Committee allocated a temporary
    group ADI of 0-20 mg/kg bw for the sucrose ester content of sucrose
    esters of fatty acids and sucroglycerides, and requested the results
    of a well designed and conducted tolerance study in humans for review
    in 1997.

         The Committee stressed that this evaluation applied to sucrose
    esters of fatty acids prepared from palmitic, stearic, and oleic
    acids, as well as palm oil, lard, and tallow. It also stressed that
    the toxicological evaluation applied only to sucrose esters of fatty
    acids as currently specified, and not to materials characterized by
    higher levels of esterification.

    5.  REFERENCES

    MITSUBISHI-KASEI INSTITUTE (1991). 13-Week oral subacute toxicity
    study of sucrose esters of fatty acids in rats. Study 0L492.
    December 27, 1991 Yokohama, Japan.

    MITSUBISHI CHEMICAL SAFETY INSTITUTE LTD. (1994a). Pharmacokinetic
    studies of sucrose esters of fatty acids (SEs) in rats, dogs and
    humans. Report no. 3B159. August 5, 1994. Yokohama, Japan.

    MITSUBISHI CHEMICAL SAFETY INSTITUTE LTD. (1994b). Clinical and
    pharmacokinetic studies of sucrose esters of fatty acids (SEs) in
    human - Supplement to report no. 3B159 - Report No 4B430 -
    Yokohama, Japan.

    MITSUBISHI CHEMICAL SAFETY INSTITUTE LTD. (1994c). Combined chronic
    oral toxicity/carcinogenicity studies of sucrose esters of fatty acids
    in rats. Report no. 1L303. October 26, 1994. Yokohama, Japan.

    MITSUBISHI CHEMICAL SAFETY INSTITUTE LTD. (1994d). Preliminary studies
    on sucrose esters of fatty acid - induced lactation. Supplement to
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    See Also:
       Toxicological Abbreviations
       Sucrose esters of fatty acids and sucroglycerides (WHO Food Additives Series 10)
       Sucrose esters of fatty acids and sucroglycerides (WHO Food Additives Series 15)
       Sucrose esters of fatty acids and sucroglycerides (WHO Food Additives Series 40)