Food Additives Permitted for Direct Addition to Food for Human Consumption; Sucrose Acetate Isobutyrate

DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Part 172

[Docket No. 91F-0228]

Food Additives Permitted for Direct Addition to Food for Human

Consumption; Sucrose Acetate Isobutyrate

AGENCY: Food and Drug Administration.

ACTION: Final rule.

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SUMMARY: The Food and Drug Administration (FDA) is amending the food

additive regulations to provide for the safe use of sucrose acetate

isobutyrate (SAIB) as a stabilizer of emulsions of flavoring oils used

in nonalcoholic beverages. This action is in response to a petition

filed by Eastman Chemical Co.

DATES: Effective June 4, 1999; written objections and requests for a

hearing by July 6, 1999. The Director of the Office of the Federal

Register approves the incorporation by reference in accordance with 5

U.S.C. 552(a) and 1 CFR part 51 of certain publications in

Sec. 172.833(b) (21 CFR 172.833(b)), effective June 4, 1999.

ADDRESS: Written objections may be sent to the Dockets Management

Branch (HFA-305), Food and Drug Administration, 5630 Fishers Lane, rm.

1061, Rockville, MD 20852.

FOR FURTHER INFORMATION CONTACT: Blondell Anderson, Center for Food

Safety and Applied Nutrition (HFS-206), Food and Drug Administration,

200 C St. SW., Washington, DC 20204, 202-418-3106.

SUPPLEMENTARY INFORMATION:

Table of Contents

I. Introduction

II. Evaluation of Safety

A. Estimated Daily Intake for SAIB

B. Evaluation of Safety Studies on SAIB

1. Pharmacokinetics and Metabolism Studies

2. Genotoxicity Studies

3. Reproduction and Developmental Toxicity Studies

a. Three-generation reproduction with teratology phase in

rats (Appendix 86)

b. Teratology study in rabbits (Appendix 87)

c. Agency conclusions regarding reproduction and

developmental toxicity studies on SAIB

4. Two-Year Carcinogenicity Studies

a. Rat study (Appendix 95)

b. Mouse study (Appendix 96)

5. Concerns Regarding Altered Liver Function

a. Liver effects in the SAIB-treated animals

i. Short-term studies

ii. Subchronic oral toxicity studies on SAIB (Appendices

63, 64, 65, 67, 68, 69, and 700

iii. Specific liver function tests

b. Studies resolving the altered liver function issue

i. One-Year chronic toxicity studies

ii. Human clinical studies

c. Agency conclusions regarding the altered liver function

issue

C. Acceptable Daily Intake for SAIB

III. Conclusion

IV. Environmental Effects

V. Paperwork Reduction Act of 1995

VI. References

VII. Objections

I. Introduction

In a notice published in the Federal Register of September 5, 1991

(56 FR 43927), FDA announced that a food additive petition (FAP 1A4266)

had been filed by Eastman Chemical Co. (Eastman), P.O. Box 511,

Kingsport, TN 37662, proposing that the food additive regulations be

amended in part 172 (21 CFR part 172) to provide for the safe use of

SAIB as a stabilizer of emulsions of flavoring oils used in

nonalcoholic carbonated and noncarbonated beverages.

SAIB is the chemical alpha-D-glucopyranoside, O-acetyl-tris-O-(2-

methyl-1-oxopropyl)-beta-D-fructofuranosyl, acetate tris(2-methyl

propanoate). It is also referred to as sucrose diacetate

hexaisobutyrate, sugar esters of fatty acids, and sucrose esters of

fatty acids.

SAIB is a slightly yellow, clear, viscous liquid, practically

odorless, with a bitter taste (not apparent at the levels used in the

regulated application). The compound is produced by reaction of food

grade sucrose with acetic anhydride and isobutyric anhydride in the

presence of a catalyst. The product is purified by molecular

distillation.

In support of safety for the proposed use of SAIB, Eastman

submitted toxicity studies performed in a variety of species. Those

studies included: Absorption, metabolism, and elimination studies

(rats, dogs, rabbits, monkeys, and humans); short-term (7 to 56 days)

studies (rats, dogs, and monkeys); a palatability study (mice);

subchronic (90 days) studies (rats and dogs); chronic studies (rats and

monkeys); carcinogenicity studies (rats and mice); reproduction studies

(rats); teratology studies (rats and rabbits); genotoxicity tests;

liver function studies (rats, dogs, monkeys, and humans); and clinical

studies (humans).

The one concern raised by FDA's evaluation of the SAIB data base

was some liver effects, which were observed in the short-term and

subchronic studies. These effects were observed primarily in SAIB-

treated dogs; for example, decreased clearance rates for

bromosulfophthalein (BSP) and indocyanine green (ICG) from the blood,

and increased serum alkaline phosphatase. To further evaluate these

liver effects, the petitioner performed special liver function tests

(BSP and ICG clearance tests) in rats, dogs, monkeys, and humans. The

BSP clearance test was also performed in monkeys and rats after

exposure to SAIB for 1 year in order to demonstrate that the liver

effects were not observed in these SAIB-

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treated animals after long-term repeated exposure. The results from

these studies and results from other studies that were pivotal to the

safety decision for the proposed use of SAIB in beverages are discussed

in section II.B of this document.

II. Evaluation of Safety

In order to establish, with reasonable certainty, that a new food

additive is not harmful under its intended conditions of use, FDA

considers the projected human dietary exposure to the additive, the

additive's toxicological data base, and other relevant information

(such as published literature) available to the agency.

A. Estimated Daily Intake for SAIB

In determining whether the proposed use of an additive is safe, FDA

typically compares an individual's estimated daily intake (EDI) of the

additive to the acceptable daily intake (ADI) established by the

toxicological database. The EDI is determined by projections based on

the amount of the additive proposed for use in particular foods and on

data regarding the consumption levels of these particular foods.

The proposed levels of use for SAIB in beverages (up to 300 parts

per million (ppm)) are supported by functionality and stability data

presented in the petition. The agency commonly uses the EDI for the

90th percentile consumer of a food additive as a measure of high

chronic exposure. For the requested food use of SAIB, the agency has

estimated the lifetime exposure for 90th percentile consumers, 2 years

old and older (all ages), to be 0.17 gram per person per day (g/p/d).

The corresponding mean intake is 0.082 g/p/d (Ref. 1).

B. Evaluation of Safety Studies on SAIB

The principal studies relevant to the safety evaluation of the

petitioned use of SAIB were performed in several animal species as

mentioned in section I of this document. The individual studies are

identified by an Appendix number in this document, as designated by

Eastman in the SAIB petition.

1. Pharmacokinetics and Metabolism Studies (Appendices 16, 17, 18, 19,

22, 23, 24, 25, 26, 27, 29, and 31)

The pharmacokinetics and metabolism studies on SAIB were performed

with rats, dogs, and humans in order to compare the absorption,

metabolism, and excretion of the food additive in animal models to that

seen in humans. Results from these studies showed the following

similarities and differences in the pharmacokinetics and metabolism of

SAIB in the test subjects:

(1) There were quantitative differences in the amounts of

administered SAIB that were absorbed by rats, dogs, and humans. Rats

and humans absorbed greater amounts of SAIB from the gastrointestinal

tract compared to dogs. In rats and humans, the majority of the orally

administered SAIB was eliminated in expired air, whereas in dogs, the

majority of SAIB was eliminated in the feces;

(2) Dogs excreted a greater proportion of the absorbed SAIB in the

bile compared to rats. The excreted materials in the bile of the dog

were identified as either unchanged SAIB or higher acylated sucrose

molecules. Lower acylated sucroses were identified in the bile of rats;

and

(3) The urinary metabolites of SAIB in rats and humans were more

similar qualitatively than those between dogs and humans. Higher

acylated sucroses were identified as the primary metabolite in the

urine of dogs. In the urine of rats and humans, only lower acylated

sucroses and free sucrose were identified. Free sucrose was not found

in the urine from dogs. These data show that more deacylation of SAIB

occurs in rats and humans than in dogs.

The patterns of absorption, metabolism, and elimination are more

similar for rats and humans than for dogs and humans. Therefore, the

agency concludes that the rat is more appropriate than the dog to model

the metabolic disposition and fate of SAIB in humans (Refs. 2, 3, 4,

and 5).

2. Genotoxicity Studies (Appendices 88, 89, 90, 91, 92, 93, and 94)

SAIB was subjected to the following battery of studies to evaluate

its genotoxic potential in prokaryotic and mammalian species: Ames

Test, Chinese Hamster Ovary Cells/HGPRT Forward Mutation Assay, In

Vitro Cytogenetic Chromosomal Aberration Assay, Unscheduled DNA Assay,

and Dominant Lethal Assay. In the absence of bioassay data, these tests

are often used to predict the carcinogenic potential of the test

compound. However, in the case of SAIB, carcinogenicity bioassays are

also available.

SAIB was shown to be nonmutagenic in the Ames test, with or without

metabolic activation (Appendices 88, 89, and 90) (Refs. 6 and 7). The

compound did not induce changes in mutation frequency in the Chinese

Hamster Ovary Cells/HGPRT Forward Mutation Assay (Appendix 91) (Ref.

8). Chromosomal aberrations were not induced in Chinese hamster ovary

cells (Appendix 92), thereby demonstrating that SAIB is not clastogenic

(Ref. 9).

Results from the Unscheduled DNA Assay (Appendix 93) were negative

regarding any significant increases in nuclear labeling or unscheduled

DNA synthesis in rat primary hepatocytes treated/incubated with SAIB

(Refs. 9, 10, 11, and 12). The Dominant Lethal Assay (Appendix 94) did

not show any significant effects on early fetal deaths per pregnancy in

rats.

Based upon the negative mutagenic and clastogenic findings in the

genotoxicity studies, the agency concludes that SAIB is not genotoxic

under the test conditions of these studies (Refs. 5, 6, 7, 8, 9, 10,

11, 12, 13, and 14).

3. Reproduction and Developmental Toxicity Studies (Appendices 86 and

87)

The objectives of the reproduction and developmental toxicity

studies were to evaluate the toxic potential of SAIB on the

reproductive system of mature rats (males and females) as well as

postnatal maturation of reproductive functions of offspring through

three successive generations. Assessment of the potential effects of

the food additive on the developing fetus was the objective of the

teratology studies.

a. Three-generation reproduction with teratology phase in rats

(Appendix 86). In this study, groups of Fischer F344 rats (three

generations: F0, F1, and F2 males and

females) were administered SAIB in the diet at dose levels of 0, 0.5,

1.0, or 2.0 g per kilogram body weight per d (g/kg bw/d). Parental

(F0) males were fed SAIB for 10 weeks prior to mating;

F0 females were fed SAIB for 2 weeks prior to mating, and

throughout mating, gestation, and lactation until the time of necropsy.

F1 and F2 males and females were exposed to SAIB

in utero; during their lactation and weaning periods as well as

throughout their mating, gestation, and lactation periods for

respective F2 and F3 litters. The F1

males and females were bred twice in succession to produce

F2a and F2b pups. For each generation, the

following reproductive parameters were examined: Mating indices,

fertility indices, gestation indices, gestation length, number of

corpora lutea, implantation efficiency, and number of early or late

resorptions. Litters from the F1 and F2

generations were examined for the number of dead pups (day 0), number

of live offspring per litter, sex ratios, pup survival percentages, pup

weights, and physical abnormalities. Macroscopic examinations of the

corpora lutea and implantations were performed on the F2

dams that were

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sacrificed on day 14 of gestation period of the F3

generation. For the teratology phase of this study, macroscopic

examination of the number and distribution of fetuses in the uterine

horn and the number of resorptions and corpora lutea were performed on

F1 dams that were sacrificed on day 20 (during gestation) of

the F2b generation. The pups from these dams were examined

for any soft-tissue or skeletal malformations.

The agency observed no reproductive or developmental toxicities in

three successive generations of rats that were exposed to SAIB at

levels up to 2.0 g/kg bw/d. There was a trend towards decreased

fertility with increasing dose of SAIB in the females of the

F1 generation during the breeding for the F2a

litters. The agency does not consider this trend to be treatment-

related because there were no significant decreases in fertility

observed in the F0 females during the breeding for the

F1 litters or the F1 females during the breeding

for the F2b litters. The agency has determined that the no

observed effect level (NOEL) for this study is 2.0 g/kg bw/d, which was

the highest dose of SAIB tested in this study (Ref. 13).

b. Teratology study in rabbits (Appendix 87). New Zealand White

SPF female rabbits were divided into a control group (32 rabbits) and 3

SAIB treatment groups (16 rabbits per group). Control and treated

female groups were induced to superovulate by receiving injections of

human chorionic gonadotropin 3 weeks prior to insemination. SAIB was

administered by oral gavage, twice daily, to the treatment groups at

dose levels of 0.50, 0.85, or 1.20 g/kg bw/d on days 7 through 19 of

gestation. The control group received only the vehicle (corn oil) .

The agency concludes that in this study there were no developmental

toxicities observed in rabbits that were exposed to SAIB by gavage at

levels up to 1.20 g/kg bw/d during gestation (days 7 to 19). The agency

has determined that the NOEL for this study is 1.20 g/kg bw/d for this

study (Ref. 13).

c. Agency conclusions regarding reproduction and developmental

toxicity studies on SAIB. Based on the data obtained from these

reproduction and developmental toxicity studies on SAIB (Appendices 86

and 87), the agency concludes that the oral administration of SAIB does

not induce reproductive or developmental effects in rats when tested in

the diet at doses up to 2.0 g/kg bw/d or developmental effects in

rabbits when tested by gavage at doses up to 1.20 g/kg bw/d. Therefore,

a NOEL of 2.0 g/kg bw/d is established for SAIB based upon the highest

dose tested in the three-generation rat study (Refs. 5 and 13).

4. Two-Year Carcinogenicity Studies (Appendices 95 and 96)

The objective of the carcinogenicity studies was to study the

carcinogenic potential of SAIB when administered to rodents for 104

weeks.

a. Rat study (Appendix 95). Fischer F344 (CDF/CrlBR) rats were

randomly assigned to 5 groups that were fed a dietary mixture of SAIB

at dose levels of 0, 0.50, 1.0, or 2.0 g/kg bw/d for 104 weeks. Two

groups of rats served as duplicate controls and were fed an NIH07 diet

that had been treated with acetone only. BW data for all of the rats

were collected on day 1, at weekly intervals during the study, and on

the day of necropsy. Food consumption was recorded weekly. Hematology

measurements were performed on all rats prior to the initiation of

treatment and at the end of the study at week 104. During necropsy,

organ weight data were collected for heart, kidneys, liver, testes,

ovaries, and brain of the rats in the two control groups and in each of

the SAIB-treated groups. Macroscopic and microscopic examinations were

performed at sacrifice (week 104) on representative tissue from a

comprehensive selection of organs from all groups of rats.

Survival in the treated rats was not significantly affected by the

SAIB treatment for the 2-year exposure duration. The antemortem changes

seen in the SAIB-treated groups at termination were similar to those

seen in the concurrent control rats and represented typical changes

seen in aging rats.

Overall, SAIB did not significantly affect the final mean bw's or

food consumption of either the male or female rats during the 104 weeks

of the study. The organ weight data showed reduced brain (absolute)

weight in the 1.0 g/kg bw/d SAIB-treated females when compared to

females in group 1 controls and increased kidney-to-brain ratios in the

1.0 g/kg bw SAIB-treated males when compared to males in group 2

controls. FDA did not consider these weight differences to be

treatment-related or toxicologically significant because they occurred

sporadically among the treated groups, that is, at only one dose level

(1.0 g/kg bw/d dose) or in only one sex. There were tumors or

nonneoplastic lesions that occurred in the control and SAIB-treated

rats of this study that represented histopathological changes commonly

seen in aging rats or represented normal variation of spontaneous tumor

incidences (e.g., testicular interstitial cell tumors, mammary gland

fibroadenomas, endometrial stromal polyps, and pituitary hyperplasia).

Thus, the histopathology data showed no evidence of male or female

SAIB-treated rats with increased incidences of tumors or nonneoplastic

lesions at any organ site that were related to the feeding of SAIB

(Ref. 15).

From this study, the agency concludes that SAIB did not induce any

tumors in Fischer 344 rats that were fed diets containing up to 2.0 g/

kg bw/d of SAIB for 104 weeks. No SAIB-related histopathological

lesions were observed in the SAIB-fed rats. Thus, the NOEL for this

study is 2.0 g/kg bw/d (Refs. 5, 14, and 15).

b. Mouse Study (Appendix 96). In this study, groups of B6C3F1/Cr1BR

mice (50 per sex per group) were fed SAIB at concentrations of 1.25,

2.5, or 5.0 g/kg bw/d in an NIH07 diet for 104 weeks. Two groups of

mice served as controls and were fed an NIH07 diet that had been

treated with acetone only. BW data were collected on day 1, at weekly

intervals during the study, and on the day of necropsy. Food

consumption was recorded weekly. Hematology measurements were performed

on mice in the control and 5.0 g/kg bw group only; 10 mice per sex

prior to the initiation of treatment and 15 mice per sex during weeks

28, 53, 79, and 105. During necropsy, organ weight data were collected

for the kidneys, liver, gall bladder, and lungs of all mice.

Macroscopic and microscopic examinations were performed at sacrifice

(week 104) on representative tissue from a comprehensive selection of

organs from all groups of mice.

The study results revealed no treatment-related effects on the

survival of SAIB-treated mice in this study. All antemortem

observations seen in the SAIB-treated mice were comparable to those

seen in the concurrent controls.

Organ-to-bw ratios of the liver and lungs of the SAIB-treated mice

were not different from the respective weight ratios in the control

mice. There were some differences in the relative kidney weights in the

SAIB-treated mice compared to controls; however, these differences were

not associated with any treatment-related kidney histopathology.

The histopathology data showed an increased incidence of SAIB-

treated male mice with bronchiolar/alveolar adenomas and an increased

incidence of SAIB-treated male mice with (combined) bronchiolar/

alveolar adenomas or carcinomas when compared to control group males

(Refs. 14 and 15). The incidences of SAIB-

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treated females with bronchiolar/alveolar adenomas or carcinomas were

comparable to incidences in control females. According to historical

control incidence data from the National Toxicology Program data base,

these incidences are within the range commonly seen in aged B6C3F1

mice. Therefore, FDA concludes that the increased incidences of SAIB-

treated mice with this tumor represent expected variations in

spontaneous incidences and were not related to the SAIB treatment

(Refs. 14 and 15). At the other organ sites, there was no evidence of

increased incidences of mice with tumors or nonneoplastic lesions that

were related to the feeding of SAIB (Refs. 14 and 15). From this study,

the agency concludes that SAIB did not induce tumors at any organ site

in B6C3F1 mice that were fed diets containing SAIB up to 5.0 g/kg bw /d

for 104 weeks. No SAIB-related nonneoplastic lesions were observed in

the SAIB-fed mice, nor was there other evidence of adverse effects in

the SAIB-fed mice at any of the tested doses. Thus, the NOEL for this

study is 5.0 g/kg bw/d (Refs. 5, 14, and 15).

5. Concerns Regarding Altered Liver Function

During the early reviews of the petition, the agency raised a

concern regarding liver effects that were observed in the SAIB-treated

animals in short-term toxicity studies (rats, dogs, and monkeys) and in

subchronic toxicity studies (rats and dogs), especially in the SAIB-

treated dogs. However, the agency could not easily determine whether

the liver effects observed in these SAIB-treated rats and monkeys were

treatment-related because of certain inadequacies in the studies, their

limited experimental designs, and the studies' short exposure

durations. These studies are discussed in section II.B.5.a of this

document. The agency also raised a concern that there were no chronic

(1 year or longer) toxicity studies on SAIB in dogs that further

examined the liver function effects.

To address these concerns, the petitioner performed BSP and ICG

clearance tests, which are specific liver function tests, with rats,

dogs, and monkeys. In addition, to address the concern regarding

possible altered liver function in chronically-exposed animals, the

petitioner performed a 1-year oral toxicity study on SAIB in monkeys;

this study included BSP clearance tests and measurements of clinical

chemistry parameters relevant to liver toxicity. The petitioner also

performed BSP clearance tests in humans that were administered doses of

SAIB up to 0.02 g/kg bw/d for 14 days to evaluate any potential effects

of SAIB on liver function in humans. These studies and the agency's

conclusions regarding them are discussed in sections II.B.5.b and

II.B.5.c of this document.

a. Liver Effects in the SAIB-treated Animals. i. Short-term Studies

(Appendices 60, 66, 71, 72, 73, 74, and 77). The following short-term

studies were designed to provide data on the short-term oral toxicity

of SAIB in rats, dogs, and monkeys with regard to potential target

organs of SAIB, as well as to determine appropriate doses for the

subchronic and chronic studies.

Rat Studies (Appendices 66 and 74). In a short-term study

(Appendix 66), SAIB was fed to groups of male and female rats at levels

of 1.0, 2.0, or 4.0 percent (equivalent to 1.0, 2.0, or 4.0 g/kg bw/d)

in the diet for 28 or 56 days. Levels of serum alkaline phosphatase

(SAP), glucose, ornithine carbamyl transferase, triglyceride,

cholesterol, and blood urea nitrogen were examined. Organ weight data

were collected only on the liver.

The limited clinical chemistry data from this study showed

decreases in blood glucose levels in female rats fed SAIB at levels of

2.0 and 4.0 percent in the diet for 56 days. The glucose levels in the

treated males were not different from comparable levels in controls for

the 56-day duration. SAP levels were not affected in the SAIB-treated

rats. There were no effects on bw or bw gain in the SAIB-treated rats.

Liver weights in these SAIB-treated rats were similar to control rats.

Also, the levels of glucose in the treated groups were not different

from controls (Ref. 16).

In another short-term study (Appendix 74), groups of rats (15 per

sex per group) were fed diets containing 0, 5,000, or 50,000 ppm

(equivalent to 0, 0.50, or 5.0 g/kg bw/d) SAIB for 3 weeks. Organ

weight data on livers from the male and female SAIB-treated rats (five

per sex per group) revealed no evidence of liver enlargement at either

of the doses of SAIB. In addition, SAIB did not affect bw gain or food

consumption in this study (Ref. 3).

Dog Study (Appendix 77). In this study (Appendix 77), six male

beagle dogs were initially fed a ground chow diet without SAIB (control

diet) daily for 3 weeks. For the next 3 to 4 weeks, the six male dogs

were fed a ground chow containing 5-percent (equivalent to 1.25 g/kg

bw/d) SAIB. ICG clearance tests were performed on four of the six dogs

at week 3 of this 5-percent SAIB feeding period. After the 3 or 4 weeks

feeding period of SAIB, the dogs were returned to control diet for an

additional 8 weeks (91st day). ICG clearance tests were performed on

week 3 and 6 of this 8-week control diet feeding period. On the 88th

day, 4 of the 6 dogs were returned to a diet containing 5-percent SAIB

for 1 day. After this 1-day SAIB feeding, SAP measurements and ICG

clearance tests were performed on the six dogs. This study did not have

a group of dogs that served as concurrent controls nor were pretest ICG

baseline values determined. Instead, the data from this study were

compared to previously reported laboratory data for ICG clearance in

normal beagle dogs.

The results of this study showed decreased clearance of serum ICG

(half-lives (t1/2)\1\ of 17.0 to 40.0 minutes) in dogs that

were fed 5-percent SAIB for 3 weeks compared to ICG clearance in normal

dogs (t1/2 of 4.2 to 8.1 minutes). ICG clearance in the

SAIB-treated dogs had returned to normal by day 84 after these dogs

were returned to control diets without SAIB. Five of six dogs had

increased SAP levels at the end of the 4-week SAIB feeding period that

were four to seven times greater than pretest values.

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\1\ Half-life(t1/2) is the time required for the

serum ICG concentrations to be reduced by one half.

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Blood glucose levels decreased (25- to 57-percent reductions) in

all of the dogs at the end of the 4-week SAIB treatment period compared

to pretest average values. However, blood glucose levels monitored at

the end of the recovery phase of the study were reversed and were

comparable to the pretest values. Ornithine carbamyl transferase and

blood cholesterol levels also increased during the SAIB exposure

period. Other blood parameters measured in these dogs (hemoglobin,

hematocrit, white blood cell counts (five out of six dogs), serum

protein, and blood urea nitrogen) were not affected by the 5-percent-

SAIB treatment. The 5-percent SAIB treatment had no effect on body

weight, food consumption, or organ weights (only liver and kidney were

measured) in the dogs for the 4-week period (Ref. 3).

Monkey Studies (Appendices 60, 71, 72, and 73). In a short-term

study (Appendix 60), SAIB was administered by oral intubation (in an

orange juice concentrate) to four monkeys (two per sex) as a single

dose that started at a dose of 1.25 g/kg bw, increased by increments of

2-fold (72-hour intervals between doses), and ended at a dose of 20 g/

kg bw over a dosing period of 14 days. All of the SAIB-dosed monkeys

survived the study. Slight to moderate watery, yellow stools were

observed in some of the monkeys administered SAIB at doses of 1.25 g/kg

bw (one male, two females), 2.5 g/kg bw (one male, one female), and 5.0

g/kg bw (one female).

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Large amounts of watery yellow stools and emesis were observed in a

monkey that received a SAIB dose of 5.0 g/kg bw dose. Gross postmortem

examinations of the four monkeys after the last dosing of SAIB revealed

no effects that were attributable to the SAIB administration (Refs. 2

and 17).

In a two-part range-finding study (Appendices 71 and 72), SAIB was

administered by oral intubation to groups of monkeys (one per sex per

group) at dose levels of 0, 0.5, 1.0, 2.0, 5.0 or 10.0 g/kg bw/d for 15

days. Incidences of soft, loose stools were observed in the SAIB-dosed

groups (1.0, 2.0, and 10.0 g/kg bw/d doses), as well as in the control

male and female groups. At the termination of the study, SAP levels in

the males of the 10.0 g/kg bw/d dose group and the females of the 5.0

and 10.0 g/kg bw/d dose groups were increased compared to their

respective controls. Pretest alkaline phosphatase levels in the SAIB-

dosed groups were also higher than pretest levels of the controls.

Decreased BSP clearance was observed in 8 out of the 10 treated

monkeys. Electron microscopy was performed only on the livers of the

control group and the high-dose group in this study. Results from the

ultrastructural analyses of the livers from the SAIB-treated monkeys

revealed increased glycogen, large glycogen aggregations surrounded by

scant smooth endoplasmic reticulum, and decreases in the amounts of

smooth endoplasmic reticulum (Refs. 2, 3, and 17). While these effects

in the SAIB-dosed monkeys suggest suppressed liver function, the agency

could not determine the toxicological significance of these effects

because of the small group sizes (Refs. 2, 3, and 17).

In another exploratory study (Appendix 73), groups of monkeys (one

per sex per group) were administered SAIB (in corn oil) orally by

gavage at doses of 0.50, 1.45, or 2.40 g/kg bw/d for 4 weeks. Control

monkeys received only the vehicle (corn oil) by gavage. BW gains were

comparable in all of the groups except for the high-dose female monkey,

who lost weight (12-percent loss) over the 4-week study duration.

Reduced food consumption was reported for this high dose female monkey.

SAP levels were increased 8-to 78-percent in the treated groups for

both sexes except for the one male in the high-dose (10 g/kg bw dose)

group. Values reported for erythrocyte counts, hemoglobin, and

hematocrits were low for all of the females in both the treatment

groups and the control group. BSP clearance rates in these monkeys were

normal. Clinical biochemistry parameters related to liver and kidney

functions were also normal in the dosed monkeys (Refs. 3 and 17).

Agency conclusions regarding short-term studies on SAIB. The

agency's overall review of the data from the preceding short-term

studies (see section II.B.5.a.i of this document) established the

following: (1) Decreased glucose levels in rats that were fed SAIB at

levels of 2.0 and 4.0 g/kg bw/d for 56 days; (2) decreased ICG

clearance rates, increased SAP levels, and decreased blood glucose

levels in dogs that were fed 1.25 g/kg bw/d SAIB for 4 weeks; and (3)

increased BSP retention and increased SAP levels in monkeys that were

administered SAIB by gavage at dose levels of 5 and 10 g/kg bw/d for 15

days. Based on these observed effects, the agency concludes that the

liver is a target organ for the toxicity of SAIB. However, because of

the short exposure durations and limited experimental designs of these

studies, the agency concludes that these studies are inadequate to

resolve concerns regarding the observed liver effects (Refs. 3 and 5).

ii. Subchronic oral toxicity studies on SAIB (Appendices 63, 64,

65, 67, 68, 69, and 70). The following subchronic oral toxicity studies

were performed in rats and dogs to examine the general systemic

toxicity of SAIB and to investigate further the liver effects of SAIB

that were observed in the short-term SAIB studies.

Rat Studies (Appendices 63, 64, and 65). In a 90-day study

(Appendix 63), groups of rats (25 per sex per group) were fed SAIB in

the diet at 0, 1, or 5 percent (equivalent to 0, 1.0, or 5.0 g/kg bw/

d). This study showed an increase (7.4 percent) in the relative liver

weights of the 5-percent SAIB-treated female rats compared to the

control females; liver weights in SAIB-treated males were not affected.

Kidney weights in the SAIB-treated groups were not different from the

kidney weights of the control rats. Final bw's were slightly decreased

(3 to 4 percent) in only the males of the 5-percent dose group. No

differences were observed in the final bw's of the males in the 1-

percent dose group or the females in all of the dose groups when

compared to respective controls. BW gain in all of the female treatment

groups was comparable to the female control groups. Overall, feed

intakes and feed efficiencies appeared to be similar across treatment

and control groups for both sexes (Ref. 16).

In another 90-day study (Appendix 64), groups of rats (10 per sex

per group) were fed SAIB in the diet at levels of 0, 0.38, 1.88, or

9.38 percent (equivalent to 0, 0.38, 1.88, or 9.40 g/kg bw/d). A slight

increase in the mean hemoglobin values and a tendency toward

leukocytosis (increased white corpuscle counts) were observed in

treated rats relative to control rats. SAP levels and BSP clearance

rates were not evaluated in this study. BW gains in the SAIB-treated

males were slightly decreased (8 to 11 percent) compared to control

males; in treated females, bw gain was not affected. Liver, kidney,

lung, gonad, spleen, and heart weights (relative and absolute weights)

of the SAIB-treated rats were not significantly different from the

respective organ weights of the control rats.

Data from the limited histopathological analyses showed an

increased incidence of clear vacuoles (fat vacuoles) in the livers of

all of the SAIB-treated rats with the greatest increase being seen in

the 1.88-percent SAIB group (Ref. 16).

In a 12-week study (Appendix 65), groups of rats (20 per sex per

group) were fed SAIB at doses of 2.5, 5.0, or 10 percent (equivalent to

2.50, 5.0, or 10.0 g/kg bw/d) in the diet. SAIB-treated male rats in

this study showed decrements in weight gain at all dose levels compared

to controls; weight gains in the SAIB-treated female rats were not

affected. There was a significant decrease in SAP levels in females

treated with 10-percent SAIB. Urinary ascorbic acid levels were

substantially decreased (47 percent in males and 64 percent in females)

in the 10-percent SAIB group relative to controls. There were no

increases in carboxyl esterase levels in any of the SAIB-treated rats.

Neither liver weights nor the ultrastructure of the livers in the SAIB-

treated rats were affected during the study. Biochemical analyses

performed on the livers of rats in the control and 10-percent SAIB

groups showed increases in liver glycogen in the 10-percent SAIB group

(in both sexes) as well as significant increases in the water content

of the livers in the males of the 10-percent SAIB group (Ref. 16).

Because of inadequacies in data analyses and reporting (e.g.,

limited statistical analyses and incomplete histopathology data) in the

subchronic rat studies, the agency could not reach a conclusion as to

whether there were treatment-related liver effects in the SAIB-fed rats

of these studies. The results from these studies did show: (1)

Significantly increased (relative to controls) relative liver weights

in rats (females only) that were fed 5-percent SAIB, and (2) increased

glycogen content and increased water content (males only) in the livers

of rats (both sexes) fed 10-percent SAIB relative to controls (Ref. 5).

[[Page 29954]]

Dog Studies (Appendices 67, 68, 69, and 70). In a 12-week study

(Appendix 67), groups of dogs (four per sex per group) were fed diets

containing 0, 0.2, 0.6, or 2.0 percent (equivalent to 0, 0.05, 0.15, or

0.5 g/kg bw/d) SAIB. This study showed increases in SAP levels in the

SAIB-treated male dogs, with a two-fold increase in the 2.0-percent

dose group. At the end of the study, relative liver weights of male and

female dogs fed SAIB at the 0.6-percent and 2.0-percent dose levels

increased compared to the respective control groups. Relative weights

of the other organs that were examined in the study (kidney, spleen,

brain, gonads, adrenals, thyroids, and pituitary) did not differ

significantly from respective relative organ weights of controls.

Survival, hematology parameters, and urine parameters tested in the

SAIB-treated dogs were also not significantly different from controls

(Ref. 18).

In another subchronic study (Appendices 68 and 69), groups of dogs

(six per sex per group) were fed dog chow containing 0-, 0.5-, 1.0-,

2.0-, or 4.0-percent (equivalent to 0, 0.13, 0.25, 0.50, or 1.0 g/kg

bw/d) SAIB for 12 weeks followed by a 3-week recovery period, during

which the dogs were fed a chow diet that did not contain SAIB. During

the 12-week treatment period and the 3-week recovery period of the

study, the control group received a basal chow meal without SAIB.

During the 12-week exposure period, all of the dogs in this study that

were fed SAIB (all doses) exhibited significant increases (3- to 7-

fold) in serum BSP concentrations compared to control dogs. BSP

retention data collected during the 3-week recovery period without SAIB

showed a reduction in BSP plasma levels in the 4-percent SAIB-treated

dogs to levels that were similar to pretest values and those seen in

control dogs (Appendix 69).

Relative liver weights increased in the male dogs fed SAIB at

levels of 1.0 and 2.0 percent in the diet; relative liver weights in

the 0.5-percent SAIB-treated males were not different from controls.

Relative liver weights in the SAIB-treated female dogs (all groups)

were not significantly different from control females. Absolute liver

weights were significantly increased in SAIB-treated males at dose

levels of 0.5, 1.0, and 2.0 percent. Liver weights of the 4.0-percent

male dose group were not analyzed at the time that the 1.0 and 2.0-

percent male dose groups were analyzed; instead, this dose group was

held for 3 additional weeks for a recovery phase of the study. At the

end of the this 3-week period (recovery phase), absolute and relative

liver weights of the 4-percent male dose group were also significantly

increased when compared to control liver weight values measured at the

end of the 12-week treatment phase. This 3-week recovery phase of the

study did not include a comparable control group of dogs that was held

for the additional 3 weeks after the treatment phase.

Data from liver biochemistry analyses showed significantly

increased liver glycogen in all of the SAIB-treated groups,

significantly increased liver lipid content in all of the dogs fed 2.0-

percent SAIB, and significantly increased liver carboxyl esterase

levels in all of the dogs fed 4.0-percent SAIB. Total protein levels in

the liver were greatly reduced in all of the SAIB-treated groups

compared to controls. Alkaline phosphatase, adenosine triphosphatase,

and glucose-6-phosphatase levels in the bile canaliculi of the livers

in all of the dose groups increased relative to controls.

Results from the microscopic (light and electron) analysis of liver

tissue samples showed dilation of the bile canaliculi, liver

hypertrophy and enlargement (males only), increased bile pigment

granules, increases in the smooth endoplasmic reticula, and prominent

Golgi bodies in the dogs fed 2-percent SAIB in the diet (Appendix 69).

In addition, the distribution and arrangement of the smooth and rough

endoplasmic reticula were altered in the 2-percent SAIB-treated dogs

(Ref. 18).

In a 91-day study (Appendix 70), a group of five dogs were fed dog

chow containing 5-percent (equivalent to 1.25 g/kg bw/d) SAIB. A second

group of five dogs served as controls and was fed dog chow containing

5-percent corn oil for the study duration. This study demonstrated that

SAIB significantly affected liver function in the five SAIB-treated

dogs, causing moderate elevations in SAP levels, prolonged ICG

clearance, and increases in the absolute and relative liver weights.

Hematological or clinical chemistry parameters examined in this study,

other than SAP, were not affected by the SAIB treatment (Ref. 3).

Based upon the data in the subchronic studies in dogs, the agency

concludes that SAIB affected liver function in SAIB-treated dogs at all

of the tested dose levels. As noted, the liver effects observed in the

SAIB-treated dogs were: (1) Increased BSP retention at SAIB doses as

low as 0.13 g/kg bw/d and up to a dose of 1.0 g/kg bw/d, (2) increased

SAP levels at SAIB doses of 0.05 g/kg bw/d and higher, (3) increased

liver weights at doses of 0.13 g/kg bw/d and higher, and (4) liver

ultrastructural changes in the 0.5 g/kg bw/d dose group (liver

enlargement/hypertrophy, increased liver glycogen deposition, increased

liver carboxyl esterase activity, and proliferation of smooth

endoplasmic reticulum). Because effects were observed at the lowest

tested dose, the agency could not establish a NOEL for the observed

liver effects in the SAIB-treated dogs in the subchronic studies (Refs.

3, 5, and 18).

Agency Conclusions Regarding Subchronic Studies on SAIB. The agency

concludes from the subchronic studies that SAIB affected liver function

in dogs when fed SAIB at doses of 0.13 g/kg bw/d up to 1.0 g/kg bw/d.

The subchronic studies in rats also suggested apparent liver

effects in rats that were fed SAIB at dose levels of 5.0 g/kg bw/d and

higher. However, because of study limitations (e.g., incomplete

histopathology data and inadequate statistical analyses), the agency

could not determine from the subchronic rat studies whether the liver

effects seen in the SAIB-treated rats were caused by the treatment with

SAIB (Refs. 3, 5, 16, and 18).

In order to investigate further the effects of SAIB on liver

function in different species, the petitioner performed specific liver

function tests in rats, dogs, monkeys, and humans. The results from

these tests are discussed in sections II.B.5.a.iii. and II.B.5.b.ii of

this document.

iii. Specific liver function tests (Appendices 75, 76, 78, 80, and

81). BSP and ICG clearance tests were performed by the petitioner in

rats, dogs, and monkeys. In these tests, BSP or ICG is administered by

injection and the clearance of these dyes from the blood is analyzed

spectrophotometrically at various time intervals up to 48 hours. In

normal subjects, generally 95 percent of the injected dye is cleared

from the blood through the liver within 30 minutes. Retention of BSP in

the blood is indicative of some form of liver dysfunction such as

hepatic degeneration/inflammatory changes, hepatic fibrosis, hepatic

cholestasis, or depressed hepatic blood flow (Refs. 19, 20, 21, 22, 23,

and 24).

Rat Tests (Appendices 75 and 76). In a 36-day study (Appendix 75),

two groups of rats (17 males per group) were fed a chow diet containing

either 4.0-percent (equivalent to 4.0 g/kg bw) SAIB in 5.0-percent corn

oil or only 5.0-percent corn oil. On days 1, 3, 5, 8, 10, 22, 26, and

36, after the start of these diets, 2 rats from each group were

selected for ICG clearance testing. ICG clearance rates in SAIB-treated

rats were not significantly different from control rats at any of the

time intervals (Ref. 3).

In a 7-day study (Appendix 76), 15 rats (5 males per group) were

fed a rodent diet containing 4-percent

[[Page 29955]]

(equivalent to 4.0 g/kg bw/d) SAIB. BSP clearance was measured in these

rats at 0, 24, and 48 hours posttreatment with SAIB. SAIB had no effect

on BSP clearance from the liver in these rats when fed for 7 days (Ref.

3).

Dog Tests (Appendices 76 and 78). In an intermittent dosing study

(Appendix 76), two male and two female dogs were serially provided, on

one dose per week, laboratory dog chow ration containing SAIB at

increasing concentrations of 0.1, 0.3, and 0.5 percent (equivalent to

dose levels of 0.03, 0.08, or 0.13 g/kg bw). The animals were fed dog

chow without SAIB on days between each dosing. BSP clearance rates for

the 4 dogs were evaluated at 24 and 48 hours following each dosing. BSP

clearance rates were also measured in each of the dogs prior to the

start of the study to determine pretest baseline values. Results from

this study showed increased BSP retention at the 24-hour time interval

in the dogs at all treatment levels (Ref. 3).

Results from another study in dogs (Appendix 78) showed that BSP

retention increased (up to seven-fold) in both male and female dogs

administered SAIB as single (oral gavage) doses ranging from 0.005 g/kg

bw to 2.0 g/kg bw. Initial increases of BSP levels were observed within

4 to 6 hours posttreatment with SAIB (Refs. 2 and 3).

Monkey Tests (Appendices 80 and 81). In a study (Appendix 80), a

group of monkeys (three males) were administered 1.0 g/kg bw of SAIB in

cottonseed oil by gavage as a single dose. A second of group of monkeys

(three males) received no treatment and served as controls. After this

dosing of SAIB, BSP clearance tests were performed. The three SAIB-

treated monkeys were given a second 1.0-g dose of SAIB after a 7-day

rest period followed by additional BSP clearance testing. The first

SAIB dosings showed an increase in the BSP level in one of the three

treated monkeys, while the second SAIB dosing resulted in an increase

in the BSP levels in a different treated monkey (Ref. 3). FDA concluded

that these results are inconclusive because of the equivocal BSP

results and the small group sizes.

In another study (Appendix 81), a group of monkeys (four males) was

administered SAIB orally by gavage at a dose of 5 g/kg bw. Another

group of four males was gavaged with corn oil and served as a control

group. BSP clearance was tested in the control and SAIB-treated monkeys

5 hours after the SAIB dosing. The group mean BSP level in the treated

monkeys was comparable to that in the control group (Ref. 3). Based

upon the results from this study, which tested a higher dose of SAIB

and had a larger group size than the above 1.0 g/kg bw monkey study

(Appendix 80), FDA concludes that BSP clearance was not affected in

monkeys that were orally gavaged with SAIB as a single dose of 1 or 5

g/kg bw (Ref. 5).

Based upon FDA's reviews of these liver function tests, the agency

concludes that liver function in dogs was clearly affected by SAIB

regardless of the doses tested (0.005 to 2.0 g/kg bw). From these

studies the agency also concludes that liver function was not affected

in either rats or monkeys at SAIB doses up to 5 g/kg bw. However,

because of the short duration of these studies, the agency was unable

to determine whether liver function would be affected in rats or

monkeys upon chronic exposure to SAIB.

In response to this concern of FDA, the petitioner conducted two 1-

year feeding studies (rats and monkeys), in which test animals were

subjected to specific liver function tests following a continuous SAIB

exposure for 1 year. Results from these 1-year studies are discussed in

section II.B.5.b.i of this document. In addition to the 1-year studies

in rats and monkeys, the petitioner conducted three human clinical

studies to investigate whether the liver function effect that was

consistently demonstrated in SAIB-treated dogs could also occur in

humans upon oral ingestion of SAIB. Results from the three human

clinical studies are also discussed in section II.B.5.b.ii of this

document.

b. Studies resolving the altered liver function issue. The

petitioner performed two 1-year chronic toxicity studies (rats and

monkeys) and the human clinical studies in an effort to resolve the

concern regarding liver function. These investigations are discussed in

sections II.B.5.b.i. and II.B.5.b.ii. of this document.

i. One-Year Chronic Toxicity Studies (Appendices 83 and 84). The

1-year chronic toxicity studies were performed in rats and in monkeys

in order to evaluate any general toxicological effects of SAIB in these

animals and to investigate whether there were effects on liver function

in rats and monkeys chronically-exposed to SAIB.

Rat Study (Appendix 83). Groups of male and female Charles River

rats (20 per sex per group) were fed SAIB in the diet at dose levels of

0, 0.5, 1.0, or 2.0 g/kg bw/d for 52 weeks. The control group was fed

the diet minus SAIB for the same duration. BSP clearance tests were

performed during weeks 23 and 48 on all control and high-dose rats

after an overnight fast. Ophthalmic examinations were performed at

weeks 0, 26, and 52 of the study. Selected hematology and clinical

chemistry tests were performed on 10 animals prior to dosing and on all

animals at weeks 27 and 53. Histopathological examinations were

performed on tissue from liver, kidneys, lungs, and all lesions from

all dose groups. Liver sections were also processed for electron

microscopy.

A small bw gain decrement (10.3 percent) was observed in the high-

dose (2.0 g/kg bw/d) SAIB-treated females. The mean final bw in the

high-dose females was also significantly decreased by 6.4 percent,

compared to controls. The decreased bw gain in the high-dose females

was mostly accounted for by decreased food intake (4-percent

reduction). BW gains in the SAIB-treated females at the mid and low

doses were not different from control females. The decreases in bw gain

that were sporadically seen in the SAIB-treated males in the short-term

studies were not observed in the males during this 1-year chronic

study. Because the bw gain decrement observed in the high-dose females

was small, and because it was not observed in either the low- or mid-

dose females or in treated males, and was partially accounted for by

decreased food intake in females, the agency concludes that this effect

is not toxicologically significant.

No differences were observed in BSP clearance between the SAIB-

treated rats and the control rats at 23 or 48 weeks. Other clinical

chemistry parameters measured in the SAIB-treated rats at week 53 were

comparable to values in control rats.

An increased incidence of high-dose female rats with hepatocellular

adenomas (2 out of 19) was observed in this study but was not seen in

the longer-term (2-year) rat carcinogenicity study on SAIB, indicating

that this effect was not treatment related (see section II.B.4 of this

document). Therefore, the agency concludes that there are no

indications of liver toxicity or other toxicologically significant

effects seen in rats chronically exposed to SAIB for 1 year. The NOEL

for this study is 2.0 g/kg bw/d, the highest dose tested (Refs. 5, 16,

and 25).

Monkey Study (Appendix 84). In this study, groups of Cynomolgus

monkeys (four per sex per group; young adults, age unknown) were

administered SAIB in corn oil by gavage at doses of 0, 0.50, 1.45, or

2.40 g/kg bw/d for 1 year. The control group was administered only corn

oil in a similar manner for the same duration. Ophthalmic,

hematological, and clinical chemistry examinations were performed at

pretest and at months 3, 6, 9, and 12 of the study. BSP clearance tests

were performed to assess liver function in all

[[Page 29956]]

animals at pretest and at months 3, 6, 9, and 12 of the study. Organ

weight data (absolute and relative) were collected on brain, thyroid/

parathyroid, heart, kidney, liver, testis, spleen, ovary, pituitary,

and adrenals for all monkeys after week 52 of the study. Macroscopic

and microscopic examinations were performed at sacrifice (week 52) on

representative tissue from a comprehensive selection of organs. Liver

sections from the monkeys in the control and high-dose group were

processed for electron microscopy.

The survival, bw, ophthalmoscopic, and hematological data showed no

findings that were toxicologically significant or SAIB-related. There

were some differences noted between the SAIB-treated and control

monkeys for some of the clinical chemistry parameters, but these were

sporadically expressed and thus were not toxicologically significant.

Data from the clinical chemistry parameters that assessed

hepatobiliary function did not reveal any effects that could be

attributed to the administration of SAIB. The percentages of BSP

excretion seen 30 minutes after BSP dye injection in the SAIB-treated

monkeys at 3, 6, and 12 months, were similar to those seen in controls

at comparable time intervals. There were no differences between SAIB-

treated monkeys and control monkeys with respect to SAP levels,

cholesterol, bile acids, bilirubin, and gamma glutamyl transpeptidase.

Organ weight data for SAIB-treated monkeys were comparable to control

monkeys except for some occasional differences in the combined weights

of the thyroid and parathyroid glands (absolute and relative) in the

low- and mid-dose male monkeys and in the absolute and relative ovary

weights in high-dose female monkeys. The liver weights (absolute or

relative weights) of the dosed monkeys were not different from the

liver weights in control monkeys. The agency concludes that none of

these changes are toxicologically significant.

Electron micrographs of liver tissue from the SAIB-treated monkeys

(high-dose group, four per sex) showed no difference from controls in

the quantity of smooth endoplasmic reticulum in their livers. Compared

to the controls, there were no ultrastructural changes in either the

mitochondria or their associated rough and smooth endoplasmic reticula

or any evidence of peroxisomal proliferation in the liver of the SAIB-

treated monkeys. Based upon these findings, the agency concludes that

there was no evidence of abnormalities in the livers of the SAIB-

treated monkeys compared to livers from control monkeys that would

indicate an SAIB-induced effect on liver function.

Based upon FDA's review of the data in this 1-year chronic study,

the agency concludes that SAIB does not affect the function or

ultrastructure of the liver in monkeys when orally administered at

doses up to 2.40 g/kg bw/d for 1 year. No other SAIB-related

histopathological lesions were observed in the SAIB-treated monkeys,

nor was there other evidence of adverse effects in the SAIB-gavaged

monkeys at any of the administered doses. Therefore, the agency has

determined that the NOEL for this study is 2.40 g/kg bw/d (Refs. 5 and

17).

ii. Human clinical studies (Appendices 97, 98, and 99). The

primary objective of the human clinical studies was to evaluate any

potential effects of SAIB on liver function in humans when administered

as a single daily dose for 14 days.

In a 14-day study (Appendix 97), SAIB was administered to 20 human

subjects (10 per sex) daily as a single dose of 0.01 g/kg bw/d. In a

second 14-day study (Appendix 98), groups of human subjects (4 per sex)

were administered daily a carbonated beverage containing SAIB at either

a dose of 0.007 g/kg bw or 0.20 g/kg bw. A third group (four per sex)

served as a control and were administered daily a carbonated beverage

without SAIB. In a third 14-day study (Appendix 99), groups of 13 human

male and 14 human female human subjects were administered daily orange

juice containing SAIB at a dose of 0.02 g/kg bw/d . In each of these

clinical studies, hematology and clinical chemistry parameters were

measured prior to the SAIB dosing on day 0, during the study on day 7,

and at the end of the study on day 14 or 18. BSP clearance tests were

performed prior to the SAIB dosing and postdosing on day 15.

None of these studies showed any SAIB-related abnormalities in any

of the hematology or clinical chemistry parameters measured in these

studies, including those clinical chemistry parameters that assessed

hepatobiliary function (i.e., SAP levels, alanine amino transferase,

aspartate amino transferase, lactate dehydrogenase, gamma glutamyl

transferase, bile acids, and total bilirubin). BSP retention in all of

the SAIB-treated human subjects was normal compared to pretest values

or control values.

Based upon the data in these studies, the agency concludes that

SAIB is not toxic in humans and does not induce liver toxicity at doses

up to 0.02 g/kg bw/day for 14 days. The 0.02 g/kg bw SAIB dose is

equivalent to exposures resulting from drinking 4 liters per day of a

beverage containing SAIB at its assumed maximum allowable use level of

300 milligrams/liter (mg/L) (Refs. 5, 26, 27, and 28).

c. Agency conclusions regarding altered liver function issue.

During the initial safety review of SAIB, FDA raised a concern that,

regardless of the tested dose or study duration, treatment-related

liver effects were consistently noted in SAIB-treated dogs. In response

to this concern, the petitioner provided a significant amount of

pharmacokinetics and metabolism data on SAIB in various species,

including humans. Based on these data, FDA finds that there appear to

be greater quantitative differences in the absorption and metabolism of

SAIB between dogs and humans than between the other tested species and

humans. To evaluate further the significance of the liver effects to

the overall safety of SAIB for human consumption, the agency carefully

considered the test results with monkeys, a nonhuman primate species

that is phylogenetically closest to humans, as well as liver function

data collected directly from human subjects in the three clinical

studies.

Unlike the liver effects seen in SAIB-treated dogs, there was no

evidence of liver effects in the specific liver function tests with

monkeys that received acute oral doses of SAIB as high as 5 g/kg bw

(Appendix 84). Data also demonstrate a lack of treatment-related liver

effects in monkeys that were exposed continuously to SAIB at dose

levels up to 2.4 g/kg bw/d over a 1-year treatment period. Importantly,

this dose level of 2.4 g/kg bw/d is nearly one thousand fold the

anticipated 90th percentile human exposure of SAIB in the daily diet.

FDA's review of the human clinical studies (Appendices 97, 98, and

99) further support the agency's conclusion regarding the significance

of the liver effects. In all three clinical studies, no SAIB-induced

effects on liver function were observed in either male or female

subjects. While the duration of the human studies was relatively short

(14 days), the highest dose used (0.02 g/kg bw/d) provided reasonable

assurance, in conjunction with the chronic monkey study data (Appendix

84), that the liver effects seen in SAIB-treated dogs will not occur in

humans that ingest SAIB. The highest dose tested in the human clinical

studies is equivalent to an exposure resulting from the drinking 4 L/d

of a beverage containing SAIB at its proposed maximum allowable use

level of 300 mg/L.

[[Page 29957]]

Based upon FDA's reviews of the nonhuman primate data and the

direct human data provided in the SAIB data base, the agency concludes

that the liver function effect seen in SAIB-treated dogs is not

determinative of the overall safety evaluation of SAIB for human

consumption. The agency further concludes that there is reasonable

certainty that the adverse liver effects seen in the SAIB-treated dogs

will not occur in humans that consume SAIB at the anticipated levels of

dietary intake.

C. Acceptable Daily Intake for SAIB

As discussed in section II.B.5.c of this document, FDA has relied

on the monkey and human data to resolve questions concerning the

altered liver function observed in SAIB-treated dogs. To support the

overall safety of SAIB for human consumption and to establish an ADI,

FDA has relied on data from rat studies of SAIB because the most

complete toxicological profile of SAIB was established in this rodent

species. The rat studies in the SAIB data base assess both the

potential carcinogenicity and the reproductive/developmental toxicity

of SAIB. In addition, because of their duration and size, the chronic

rat studies had greater sensitivity and thus, were more likely to

manifest treatment-related chronic effects. Furthermore, the available

absorption and metabolism data demonstrated substantial similarities,

both qualitative and quantitative, between rats and humans in the

metabolic handling of SAIB following oral ingestion.

Based on the 1- and 2-year rat studies, FDA determined that the

highest dose tested in both studies (2.0 g/kg bw/d) was the NOEL for

SAIB. Based on this NOEL and the use of a safety factor of 100, FDA

calculated an ADI of 0.02 g/kg bw/d or 1.20 g/p/d for SAIB (Ref. 5).

The EDI exposure for SAIB is 0.17 g/p/d (90th percentile, all ages)

which is 14 percent of the ADI calculated for the additive.

III. Conclusion

Based on all the SAIB data reviewed by the agency, FDA concludes

that there is a reasonable certainty that no harm will result from the

use of SAIB as an emulsion stabilizer for flavoring oils in

nonalcoholic beverages, and thus, SAIB is safe for its proposed use.

Therefore, the agency concludes that the food additive regulations

should be amended as set forth in this document.

In accordance with Sec. 171.1(h) (21 CFR 171.1(h)), the petition

and the documents that FDA considered and relied upon in reaching its

decision to approve the petition are available for inspection at the

Center for Food Safety and Applied Nutrition by appointment with the

information contact person listed above. As provided in Sec. 171.1(h),

the agency will delete from the documents any materials that are not

available for public disclosure before making the documents available

for inspection.

IV. Environmental Effects

The agency has determined under 21 CFR 25.32(k) that this action is

of a type that does not individually or cumulatively have a significant

effect on the human environment. Therefore, neither an environmental

assessment nor an environmental impact statement is required.

V. Paperwork Reduction Act of 1995

This final rule contains no collection of information. Therefore,

clearance by the Office of Management and Budget under the Paperwork

Reduction Act of 1995 is not required.

VI. References

The following references have been placed on display in the Dockets

Management Branch (address above) and may be seen by interested persons

between 9 a.m. and 4 p.m., Monday through Friday. Note: References with

an asterisk are not on display; they are available generally because

they are published articles or books.

1. Memorandum, from DiNovi, Division of Product Manufacture and

Use, to Anderson, Direct Additives Branch, FDA, July 31, 1991.

2. Memorandum from Taylor, Additives Evaluation Branch, to

McLaughlin, Direct Additives Branch, FDA, November 4, 1985.

3. Memorandum from Pellicore, Additives Evaluation Branch, to

Anderson, Direct Additives Branch, FDA, September 29, 1992.

4. Memorandum from Pellicore, Additives Evaluation Branch, to

Anderson, Novel Ingredients Branch, FDA, February 5, 1993.

5. Memorandum from Whiteside, Division of Health Effects

Evaluation, to Anderson, Division of Product Policy, FDA, August 27,

1998.

6. Memorandum from Donnely, Genetic Toxicity Branch, to Dunkel,

Genetic Toxicity Branch, FDA, September 23, 1985.

7. Memorandum from Prival, Additives Evaluation Branch #1, to

Whiteside, Additives Evaluations Branch #2, FDA, March 14, 1996.

8. Memorandum from Moreland, Genetic Toxicity Branch, FDA,

August 12, 1985.

9. Memorandum from Lavappa, Genetic Toxicity Branch, to Chief,

Genetic Toxicity Branch, FDA, September 9, 1985.

10. Memorandum from Bradlaw, Genetic Toxicity Branch, to Dunkel,

Genetic Toxicity Branch, FDA, May 24, 1985.

11. Memorandum from Bradlaw, Genetic Toxicity Branch, to Dunkel,

Genetic Toxicity Branch, FDA, August 15, 1985.

12. Memorandum from Bradlaw, Genetic Toxicity Branch, to Lin,

Additives Evaluation Branch, FDA, September 6, 1989.

13. Memorandum from Welsh, Additives Evaluation Branch #2, to

Whiteside, Additives Evaluations Branch #2, FDA, August 28, 1995.

14. Memorandum from Whiteside, Division of Health Effects

Evaluation, to Lorentzen, Cancer Assessment Committee, FDA, March

18, 1997.

15. Memorandum of Conference, Cancer Assessment Committee

Meeting, FDA, October 28, 1996.

16. Memorandum from Raffaele, Additives Evaluations Branch #2,

to Whiteside, Additives Evaluations Branch #2, FDA, June 23, 1994.

17. Memorandum from Whiteside, Additives Evaluation Branch #2,

to Anderson, Direct Additives Branch, FDA, March 23, 1994.

18. Memorandum from Whiteside, Additives Evaluation Branch #2,

to Anderson, Direct Additives Branch, FDA, August 20, 1993.

*19. Cornelius, C. E., ``Liver Function,'' Clinical

Biochemistry of Domestic Animals, C. E. Cornelius and J. J. Kaneko,

eds. Academic Press, pp. 251-264, 1963.

*20. Cornelius, C. E., et al., ``An assessment of hepatic

function in rhesus and squirrel monkeys,'' Veterinary Medicine/Small

Animal Clinicals, 78: 1885-1888, 1983.

*21. Cornelius, C. E., ``Liver Function,'' Clinical Biochemistry

of Domestic Animals, 4th ed. Jiro J. Kaneko, Ed., Academic Press,

Inc., pp. 375-379, 391-397,1989.

*22. Poutsiaka, et al., ``Simultaneous Determination in Dogs of

Liver and Kidney Functions with Bromosulfalein and

Phenolsulfonephthalein,'' Toxicology and Applied Pharmacology, 4:55,

1962.

*23. Krasavage, et al., ``Indocyanine Green Testing,''

Proceedings Society for Experimental Biology of Medicine, 119: 215,

1965.

*24. Bonasch, H. and C. E. Cornelius, ``Indocyanine Green: A

Liver Function Test for the Dog,'' American Journal of Veterinary

Research, 25: 254-59, 1964.

25. Memorandum from Alam, Pathology Branch, to Anderson, Novel

Ingredients Branch, FDA, July 10, 1997.

26. Memorandum from Hotta, Clinical Nutrition Branch, to

Blendermann, Division of Nutrition, FDA, December 31, 1986.

27. Memorandum from Calvert, Clinical Nutrition Branch, to

Anderson, Novel Ingredients Branch, FDA, January 21, 1992.

28. Memorandum from Calvert, Clinical Nutrition Branch, to

Anderson, Novel Ingredients Branch, FDA, September 2, 1993.

VI. Objections

Any person who will be adversely affected by this regulation may at

any time on or before July 6, 1999, file with the Dockets Management

Branch (address above) written objections thereto. Each objection shall

be separately numbered, and each numbered objection shall specify with

particularity the provisions of the regulation to which objection is

made and the grounds for the objection. Each numbered objection on

which a hearing is requested shall specifically so state.

[[Page 29958]]

Failure to request a hearing for any particular objection shall

constitute a waiver of the right to a hearing on that objection. Each

numbered objection for which a hearing is requested shall include a

detailed description and analysis of the specific factual information

intended to be presented in support of the objection in the event that

a hearing is held. Failure to include such a description and analysis

for any particular objection shall constitute a waiver of the right to

a hearing on the objection. Three copies of all documents shall be

submitted and shall be identified with the docket number found in

brackets in the heading of this document. Any objections received in

response to the regulation may be seen in the Dockets Management Branch

between 9 a.m. and 4 p.m., Monday through Friday.

List of Subjects in 21 CFR Part 172

Food additives, Incorporation by reference, Reporting and

recordkeeping requirements.

Therefore, under the Federal Food, Drug, and Cosmetic Act and under

authority delegated to the Commissioner of Food and Drugs, 21 CFR part

172 is amended as follows:

PART 172--FOOD ADDITIVES PERMITTED FOR DIRECT ADDITION TO FOOD FOR

HUMAN CONSUMPTION

1. The authority citation for 21 CFR part 172 continues to read as

follows:

Authority: 21 U.S.C. 321, 341, 342, 348, 371, 379e.

2. Section 172.833 is added to subpart I to read as follows:

Sec. 172.833 Sucrose acetate isobutyrate (SAIB).

Sucrose acetate isobutyrate may be safely used in foods in

accordance with the following prescribed conditions:

(a) Sucrose acetate isobutyrate (CAS Reg. No. 27216-37-1), or SAIB,

is the chemical alpha-D-glucopyranoside, O-acetyl-tris-O-(2-methyl-1-

oxopropyl)-beta-D-fructofuranosyl, acetate tris(2-methyl propanoate).

(b) SAIB, a pale, straw-colored liquid, meets the following

specifications:

(1) Assay: Not less than 98.8 percent and not more than 101.9

percent, based on the following formula:

Assay = ((SV 0.10586) 56.1) x 100

Where SV = Saponification value

(2) Saponification value: 524-540 determined using 1 gram of sample

by the ``Guide to Specifications for General Notices, General

Analytical Techniques, Identification Tests, Test Solutions, and Other

Reference Materials,'' in the ``Compendium of Food Additive

Specifications, Addendum 4, Food and Agriculture Organization of the

United Nations (FAO), Food and Nutrition Paper 5, Revision 2'' (1991),

pp. 203 and 204, which is incorporated by reference, in accordance with

5 U.S.C. 552(a) and 1 CFR part 51. Copies are available from the Office

of Premarket Approval, Center for Food Safety and Applied Nutrition

(HFS-200), Food and Drug Administration, 200 C St. SW., Washington, DC

20204, or may be examined at the Center for Food Safety and Applied

Nutrition's Library, 200 C St. SW., rm. 3321, Washington, DC, or at the

Office of the Federal Register, 800 North Capitol St. NW., suite 700,

Washington, DC.

(3) Acid value: Not to exceed 0.20 determined using 50 grams of

sample by the ``Guide to Specifications for General Notices, General

Analytical Techniques, Identification Tests, Test Solutions, and Other

Reference Materials,'' in the ``Compendium of Food Additive

Specifications, Addendum 4, FAO Food and Nutrition Paper 5, Revision

2,'' p. 189 (1991), which is incorporated by reference; see paragraph

(b)(2) of this section for availability of the incorporation by

reference.

(4) Lead: Not to exceed 1.0 milligrams/kilogram determined by the

``Atomic Absorption Spectrophotometric Graphite Furnace Method, Method

I,'' in the ``Food Chemicals Codex,'' 4th ed. (1996), pp. 763 and 764,

with an attached modification to the sample digestion section in

Appendix III.B (July 1996), which is incorporated by reference. Copies

are available from the National Academy Press, 2101 Constitution Ave.

NW., Box 285, Washington, DC 20055 (Internet ``http://www.nap.edu''),

or may be examined at the Center for Food Safety and Applied

Nutrition's Library, 200 C St. SW., rm. 3321, Washington, DC, or at the

Office of the Federal Register, 800 North Capitol St. NW., suite 700,

Washington, DC.

(5) Triacetin: Not to exceed 0.10 percent determined by gas

chromatography as described in the ``Guide to Specifications for

General Notices, General Analytical Techniques, Identification Tests,

Test Solutions, and Other Reference Materials,'' in the ``Compendium of

Food Additive Specifications, Addendum 4, FAO Food and Nutrition Paper

5, Revision 2'' (1991), pp. 13-26, which is incorporated by reference;

see paragraph (b)(2) of this section for availability of the

incorporation by reference.

(c) The food additive is used as a stabilizer (as defined in

Sec. 170.3(o)(8) of this chapter) of emulsions of flavoring oils in

nonalcoholic beverages.

(d) The total SAIB content of a beverage containing the additive

does not exceed 300 milligrams/kilogram of the finished beverage.

Dated: May 27, 1999.

William K. Hubbard,

Associate Commissioner for Policy Coordination.

[FR Doc. 99-14147 Filed 6-3-99; 8:45 am]

BILLING CODE 4160-01-F