[Federal Register Volume 72, Number 51 (Friday, March 16, 2007)]
[Rules and Regulations]
[Pages 12560-12564]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: E7-4796]


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DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Part 584

[Docket No. 1995G-0321] (formerly 95G-0321)


Food Substances Affirmed as Generally Recognized as Safe in Feed 
and Drinking Water of Animals: 25-Hydroxyvitamin D3

AGENCY: Food and Drug Administration, HHS.

ACTION: Final rule.

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SUMMARY: The Food and Drug Administration (FDA) is amending its 
regulations to affirm that the use of 25-hydroxyvitamin D3 
is generally recognized as safe (GRAS) as a source of vitamin 
D3 activity in broiler chicken feeds and drinking water when 
used in accordance with certain limitations. This action is in response 
to a petition filed by Amoco BioProducts Corp. Subsequently, the 
sponsorship for this petition was changed to IsoGen L.L.C., Monsanto 
Co., Roche Vitamins, Inc., and lastly, to DSM Nutritional Products, 
Inc.

DATES: This rule is effective March 16, 2007.

FOR FURTHER INFORMATION CONTACT: Michaela Alewynse, Center for 
Veterinary Medicine (HFV-228), Food and Drug Administration, 7519 
Standish Pl., Rockville, MD 20855, 240-453-6866, e-mail: 
[email protected].

SUPPLEMENTARY INFORMATION:

I. Background

    In accordance with the procedures described in 21 CFR 570.35, Amoco 
BioProducts Corp., P.O. Box 3011, Naperville, IL, 60566, submitted a 
petition (GRASP 2449) requesting that 25-hydroxyvitamin D3 
(25-OH D3) be affirmed as GRAS for use as a source of 
vitamin D3 activity in broiler chicken feeds. In the 
original petition, 25-OH D3 was proposed for use in feed 
only. The proposed use was amended in a submission dated January 7, 
1998, to include administration through drinking water. Furthermore, 
all data for feed are applicable to water.
    FDA published a notice of filing of this petition in the Federal 
Register of October 24, 1995 (60 FR 54505), and gave interested parties 
an opportunity to submit comments to the agency. FDA did not receive 
any comments in response to that notice. Subsequent to the filing of 
the petition, sponsorship was changed to IsoGen L.L.C., Monsanto Co., 
Roche Vitamins, Inc., and lastly, to DSM Nutritional Products, Inc., 45 
Waterview Blvd., Parsippany, NJ, 07054-1298.

II. Standards for GRAS Affirmation

    Under Sec.  570.30 (21 CFR 570.30), general recognition of safety 
of food ingredients may be based only on the views of experts qualified 
by scientific training and experience to evaluate the safety of food 
substances directly or indirectly added to food. The basis of such 
views may be either of the following: (1) Scientific procedures, or (2) 
in the case of a substance used in food prior to January 1, 1958, 
through experience based on common use in food. General recognition of 
safety based upon scientific procedures requires the same quantity and 
quality of scientific evidence as is required to obtain approval of a 
food additive regulation for the ingredient and ordinarily is to be 
based upon published studies, which may be corroborated by unpublished 
studies and other data and information (Sec.  570.30(b)). General 
recognition of safety through experience based on common use of a 
substance in food prior to January 1, 1958, may be determined without 
the quantity or quality of scientific evidence required for approval of 
a food additive regulation. Ordinarily it is to be based upon generally 
available data and information (Sec.  570.30(c)).
    The subject petition relies on scientific procedures evidence to 
support the GRAS affirmation of 25-OH D3 as a source of 
vitamin D3 activity in broiler chicken feeds and drinking 
water.

III. Safety Evaluation

A. Introduction

    25-OH D3, also called 25-hydroxycholecalciferol, is a 
normal metabolite of vitamin D3 in mammals and birds. 
Chemically, the substance is 9,10-secocholesta-5,7,10(19)-triene-
3[beta], 25-diol. 25-OH D3 is the principal circulating form 
of vitamin D3, which is the primary source of vitamin D 
activity for livestock animals. The metabolism of vitamin D in animals 
is well understood and is documented in biochemistry textbooks (for 
example, Ref. 1). In poultry, vitamin D regulates calcium and 
phosphorus homeostasis, bone growth, eggshell formation, as well as 
other endocrine system functions (Ref. 2).
    Animals, including poultry, do not have a dietary requirement for 
vitamin D when sufficient ultraviolet (UV) light is available, because 
vitamin D is produced through action of UV light on a provitamin 
present in the skin. This provitamin is synthesized in the body and 
present in large amounts in skin, intestinal wall, and other tissues 
(Ref. 2). Vitamin D becomes a nutritionally important factor in the 
absence of sufficient UV light either from the sun or from an 
artificial source. Under modern farming conditions, many animals are 
raised in total confinement with limited exposure to UV light thus 
creating the need for a dietary supply of vitamin D.
    There are two predominant forms of vitamin D for poultry. Vitamin 
D2 comes mainly from plants. Vitamin D3 is 
produced in a bird's body when sunlight reacts with vitamin D 
precursors obtained from the bird's diet. Since vitamin D3 
is 30 to 40 times more potent than D2, plants are considered 
insignificant sources of vitamin D for birds.
    Commonly, broiler chickens are grown within the confines of 
buildings with large numbers of birds per building and are supplied 
with bulk feed and water for ad libitum consumption. Various strains of 
chicken have been developed for broiler production. They have been bred 
primarily for rapid weight gain and efficient feed utilization. 
Typically, broilers are slaughtered at 6 to 7 weeks of age if size and 
weight requirements are attained. Crumbled starter feed is supplied 
during weeks 1 to 3, pelletized grower feed during weeks 4 to 6, and 
finisher feed until slaughter. The major differences among these types 
of feed are the levels and sources of nutrients provided in the feed, 
such as amino acids, minerals, and vitamins. The level of vitamin 
supplementation provided in the broiler industry is based on type of 
diets fed, species, age of the bird, dietary antagonists, form of 
vitamin product, requirement status (optimum or minimum requirements), 
disease status, complexity of the ration, and environmental factors, 
primarily ambient temperature. Only after all these factors are 
considered can the optimal vitamin requirements for poultry be 
estimated (Ref. 2).
    The National Research Council's (NRC) recommendation for dietary 
vitamin D3 requirement of broiler chickens is 200 
International Units (IU) of vitamin D3 per kilogram (/kg) of 
feed (Ref. 3). One unit of vitamin D3 is

[[Page 12561]]

defined as the activity of 0.025 microgram ([micro]g) of vitamin 
D3. Thus, a supplement of 200 IU/kg of feed is equivalent to 
5 [micro]g of vitamin D3/kg of feed. This requirement is 
based on diets containing the required amounts of calcium and available 
phosphorus and is considered by NRC to be the minimum amount required 
to prevent deficiency signs.

B. Manufacturing and Specifications

    According to the petition, the production of 25-OH D3 
uses a bioengineered strain of the yeast Saccharomyces cerevisiae. The 
25-OH D3 final product is a white to slightly pink, 
odorless, crystalline substance. The petition lists the specifications 
for 25-OH D3 as: not less than 94.0 percent 25-OH 
D3; not more than 1 percent of any individual sterol; not 
more than 5 percent water; not more than 20 parts per million (ppm) 
lead; not more than 20 ppm aluminum; not more than 1.0 percent 
solvents; and non-detectable levels of 2', 4', 5', 7'-
tetraiodofluorescin. In order to ensure vitamin potency so that the 25-
OH D3 ``performs an appropriate function in the food,'' an 
expiration date should be included on feed and water premixes (Sec.  
570.30(f)(2)).

C. Use in Feed and Drinking Water

    The petitioner claims that the NRC recommendation of 5 [micro]g of 
vitamin D3/kg of feed is virtually never used in the broiler 
industry because commercial broiler strains currently grow much faster, 
utilize feed more efficiently, and are reared in confinement with less 
exposure to UV light than when the NRC made its recommendation. As a 
result, petitioner claims supplementation of broiler feed with vitamin 
D3 is typically at a considerably higher level. A survey of 
commercial practices in regard to vitamin supplementation of poultry 
feed supported this argument, i.e., it revealed that the amounts of 
vitamin D3 commonly added by the broiler chicken feed 
companies range from 50.0 to 62.5 parts per billion (ppb) ([micro]g/kg) 
of finished feed (Ref. 4).
    The petitioner proposes that 25-OH D3is GRAS when added 
to broiler chicken feed at levels not to exceed 69 ppb ([micro]g/kg) of 
finished feed. Based on the manufacturing and composition of a liquid 
product and its liquid release and stability data, the petitioner also 
proposes that 25-OH D3 is GRAS when added to broiler 
chickens' drinking water at levels not to exceed 34.5 ppb. This is 
because it is generally assumed that birds drink approximately twice as 
much water as the amount of feed consumed on a weight basis. To assure 
safe use of 25-OH D3, the label and labeling shall bear 
adequate mixing directions to ensure that the product (and its 
premixes) is uniformly blended throughout the feed or drinking water. 
In addition, since there are no animal consumption data to support the 
concurrent use of 25-OH D3 in feed and water, there must be 
a statement on all premix labeling (feed and drinking water forms) that 
25-OH D3 should not be used concurrently in both feed and 
water.

D. General Recognition of Safety

    The petition provides information to support a determination that 
the use of 25-OH D3 in broiler chicken diets or drinking 
water is GRAS based upon the existence of an expert consensus, based on 
scientific procedures, that 25-OH D3 has been shown to be 
safe. Foremost in the support of the determination is the same kind and 
quality of safety data as would be required to obtain FDA approval of 
25-OH D3 for use as a food additive. In particular, the 
majority of the data is published, and there is a consensus among 
qualified experts, based on the data, that this use of the substance is 
safe.
    Information in the petition shows that the safety of 25-OH 
D3 has been evaluated by an expert panel. The expert panel 
was convened by the Life Sciences Research Office, Federation of 
American Societies for Experimental Biology. The expert panel obtained 
background information, identified and analyzed pertinent literature 
and experimental studies, and reached an opinion as to whether the 
available information and data on the health effects of 25-OH 
D3 were sufficient to meet the regulatory requirements of 
safety as a GRAS substance for the intended use. The expert panel 
concluded that the available information supports a GRAS classification 
of 25-OH D3 when supplied as a source of vitamin D activity 
in broiler feed at the intended level of use of about 69 [micro]g/kg of 
feed. Corroborating evidence has shown that 25-OH D3 is a 
normal metabolite of vitamin D3 and the principal 
circulating form of vitamin D3 in mammals and birds (Ref. 
2). In addition, the petitioner provided testimony of a world-renowned 
expert on vitamin D who concluded that 25-OH D3 would be a 
safer dietary ingredient than vitamin D, since it does not accumulate 
in the body and thus, would not cause toxicity because of accumulation 
to toxic levels (Ref. 5).
1. Target Animal Safety
    The NRC reported that the existing data for broiler chickens do not 
allow precise estimates to be made for maximum vitamin D (and vitamin D 
metabolites such as 25-OH D3) tolerance levels; however, it 
indicated that under short-term feeding conditions (less than 60 days), 
most species including chickens, can tolerate as much as 100 times (100 
X) their apparent vitamin D dietary requirements, i.e., 500 [micro]g/kg 
of feed (Ref. 6).
    Estimates of the tolerance of 25-OH D3 by broiler 
chickens were assessed by the expert panel (Ref. 7) primarily from two 
field trials conducted for Amoco BioProducts Corp. The aim of these 
studies was to determine the utility of 25-OH D3 as a source 
of vitamin D3 activity and/or to evaluate whether 25-OH 
D3 exhibited toxic effects when added to broiler diets. One 
of the studies (Ref. 8) was compromised due to the high mortality rate 
(up to 16 percent) that occurred in all groups including controls. The 
other study (Ref. 9) was requested by the expert panel and is discussed 
below.
    In addition to the field trials conducted for Amoco BioProducts 
Corp., the expert panel evaluated information from separate published 
sources, including the results of several animal feeding studies. Based 
on its comprehensive review of the literature, the expert panel 
concluded that dietary levels up to 10 [micro]g of 25-OH D3/
kg of feed are safe for broiler chickens for prolonged feeding. It was 
noted that broiler chickens fed 100 [micro]g of 25-OH D3/kg 
of feed exhibited toxicity signs characterized by epithelial necrosis 
and mineralization in the distal convoluted tubules of the kidney (Ref. 
10). However, a detailed evaluation of the study revealed significant 
inadequacies in its design and experimental procedures. Thus, the 
expert panel did not place great weight on this study in its evaluation 
of safety. Additionally, a number of studies were performed in other 
poultry species. No signs of toxicity were reported in laying hens when 
25-OH D3 was fed at levels up to 50 [micro]g/kg of feed for 
periods up to 448 days (Ref. 11). Laying Japanese quail and growing 
turkeys were fed diets containing up to 16.6 [micro]g of 25-OH 
D3/kg of feed for periods up to 42 days with no adverse 
effects recorded (Refs. 12 and 13).
    Because the concentration of 25-OH D3 in broiler feed 
that may elicit toxic effects was not known, the expert panel requested 
that Amoco BioProducts Corp. conduct an additional target animal safety 
study (Ref. 9). 25-OH D3

[[Page 12562]]

was tested at multiple levels at and above (0, 1, 10, 50, 100, and 
200X) the maximum proposed use level. No significant differences in 
body weight, mortality rate, or treatment-related lesions were observed 
in the 25-OH D3 group at the maximum proposed level of 69 
[micro]g/kg of feed. However, 35 percent of the birds in the 690 
[micro]g 25-OH D3/kg group (10X) developed renal 
calcification. In addition, by the end of the study, the body weights 
of the birds from the 690 [micro]g/kg group fell 12 percent when 
compared with the birds from the 69 [micro]g/kg group. Data were not 
collected to determine the level between 69 and 690 [micro]g 25-OH 
D3/kg of feed at which toxic effects were seen. High 
mortality was observed in birds exposed to high levels of 25-OH 
D3 (50, 100, and 200X) thus, these treatments were 
terminated before the end of the study. Based on its review of the 
available information, including the results of this additional study, 
the expert panel concluded that the proposed maximum level (69 [micro]g 
25-OH D3/kg of feed) is within the range that growing 
broilers can tolerate.
    After evaluating the results of the latter study (Ref. 9), FDA 
found that, although no toxic effects were observed at the maximum 
proposed use level (69 [micro]g/kg of feed), significant treatment-
related lesions occurred when birds were exposed to 10 times (690 
[micro]g/kg of feed) that level. In the absence of key data to 
determine the level between 69 and 690 [micro]g/kg at which toxicity 
effects were seen, there was no assurance that toxicity would not occur 
at levels just above 69 [micro]g/kg. Thus, to identify the margin of 
safety for the intended use, FDA requested another target animal safety 
study.
    The study (Ref. 14) was designed to show safety to broiler chickens 
by testing 25-OH D3 at varying levels between 69 and 690 
[micro]g/kg of feed which represented 0 (control), 1, 3, 5, and 10 
times the maximum proposed use level. The results of the study 
corroborated previous findings that 25-OH D3 supplementation 
at levels up to the maximum proposed use rate resulted in no toxic 
effect, and at levels 10 times the proposed use level resulted in renal 
calcification. The study also found that no treatment-related lesions 
occurred when broilers were fed at levels up to five times the highest 
proposed use level.
    FDA has determined that the published studies, as corroborated by 
this unpublished study, provide an adequate basis upon which to 
conclude that 25-OH D3 is a safe source of vitamin 
D3 activity for broiler chickens when fed at nutritional 
levels not to exceed 69 [micro]g/kg of finished feed.
2. Consumer Exposure
    The safety of 25-OH D3 has been evaluated to assess its 
potential toxicity in humans who consume edible tissues derived from 
broiler chickens fed 25-OH D3--supplemented feed. The 
evaluation was based on FDA's review of published and unpublished 
information provided in the petition, including the safety evaluation 
performed by the expert panel (Ref. 7). Based on the available 
information and evaluation of the biological effects of 25-OH 
D3, the expert panel concluded that ``The available 
information supports a Generally Recognized as Safe (GRAS) 
classification of 25-hydroxyvitamin D3 when supplied as a 
source of vitamin D activity in broiler feed at the intended level of 
use of about 68.8 [micro]g (63.8 to 73.7 [micro]g) per kilogram of 
feed.''
    Having evaluated the data and information contained in the 
petition, FDA preliminarily found that this use of 25-OH D3 
in broiler feed was safe; however, some concerns regarding the 
teratogenicity of 25-OH D3 in the rabbit (the most sensitive 
species) remained. Specifically, Dutch Belted rabbits dosed at 25 and 
50 [micro]g/kg bodyweight (BW)/day by oral intubation from gestation 
day 6 to day 18 produced pups with skeletal and vascular anomalies. 
These dose-related effects included domed skulls, enlarged cardiac 
atria, and dilated pulmonary arteries. None of these abnormalities were 
noted in the negative controls or the 5 [micro]g/kg BW/day group. This 
and other toxicity studies were previously reviewed by FDA in support 
of the use of 25-OH D3 as the therapeutic drug for humans, 
calcifediol (Ref. 15).
    The petitioner argued that the rabbit's unique calcium metabolism 
and unusual sensitivity to the effects of vitamin D compounds rendered 
it too sensitive a model for assessing the teratogenic potential of 
vitamin D in humans (Ref. 16). In addition, the petitioner provided the 
testimony of Dr. Hector De Luca, a world-renowned expert (Ref. 5) on 
vitamin D, who noted that rabbits are extraordinarily sensitive to 
vitamin D and that they rapidly go into hyperglycemia at low doses of 
any form of vitamin D. Dr. De Luca stated that in the rabbit, 
hyperglycemia likely causes the observed teratogenic effects. In the 
expert's view, the rabbit should not be used as a toxicology model for 
man for studies of vitamin D compounds, because they almost stand alone 
in their sensitivity among species used for the safety studies.
    Based on information provided in the petition, FDA concurs that the 
rabbit is unusually sensitive to the effects of vitamin D compounds. 
However, the agency does not have sufficient information to disqualify 
the rabbit model in toxicity testing. While not disregarding the rabbit 
study, FDA took into account the high sensitivity of the rabbit model 
and used a 100-fold safety factor rather than the usual 1000-fold 
safety factor in calculating an acceptable daily intake (ADI) for 25-OH 
D3. Consequently, FDA set the ADI based on current 
information at 0.05 [micro]g/kg BW/day. The safe concentrations of 25-
OH D3 in chicken tissues based on an ADI of 0.05 [micro]g/kg 
BW/day is 10 ppb in muscle, 30 ppb in liver, and 60 ppb in skin/fat. 
These values are well within the estimated safe concentrations for 
consumers (Ref. 17). Although liver concentrations of 25-OH 
D3 were not measured, it is anticipated that these values 
similarly would be within the calculated acceptable levels. FDA 
concludes that the available data indicate that residue levels of 25-OH 
D3 will not result in any unsafe residues of 25-OH 
D3 in edible chicken tissues.
    In its evaluation of 25-OH D3, FDA has reviewed not only 
the safety of the product itself, but also the safety of the chemical 
impurities that may be present in the product from the manufacturing 
process. Residual amounts of reactants and manufacturing aids are 
commonly found as contaminants in chemical products, including products 
added to animal feeds.
    The biological stain, 2', 4', 5', 7'-tetraiodofluorescin (also 
known as FD&C Red No. 3 or erythrosin), is used as a photosensitizer in 
the production of 25-OH D3. This use of 2', 4', 5', 7'-
tetraiodofluorescin may result in unintended residue levels of 50 ppm 
maximum in the finished 25-OH D3 product (Ref. 18). At a 50 
ppm concentration in the 25-OH D3 final product, when the 
25-OH D3 is diluted in feed to a concentration of 69 ppb (69 
[micro]g/kg), 2', 4', 5', 7'-tetraiodofluorescin would be present at 
3.45 parts per trillion (ppt) in finished broiler chicken feed.
    FDA used risk assessment procedures to estimate the upper-bound of 
risk presented by 2', 4', 5', 7'-tetraiodofluorescin, a carcinogenic 
chemical (21 CFR 81.10(u)), that may be present in the 25-OH 
D3 product. Using a worst case estimate, taking into account 
a bird feed efficiency of approximately 1.9 and 80 percent uptake, the 
worst case concentration of 2', 4', 5', 7'-tetraiodofluorescin in 
edible tissue would be 5.2 ppt (3.45 ppt x 1.9 x 0.8 = 5.2 ppt). 
Therefore, the agency would not expect this impurity to become a 
component of food at other than minute levels. However, because

[[Page 12563]]

2', 4', 5', 7'-tetraiodofluorescin is a carcinogenic chemical, FDA 
advised that the sponsor should either find a replacement for 2', 4', 
5', 7'-tetraiodofluorescin or remove 2', 4', 5', 7'-tetraiodofluorescin 
from the final product in a consistent manner. Subsequently, the 
petitioner developed a purification process for complete removal of 2', 
4', 5', 7'-tetraiodofluorescin from the final 25-OH D3 
product. The petitioner provided details on the experimental conditions 
and the supporting analytical data for the analytical method used to 
quantitate residual 2', 4', 5', 7'-tetraiodofluorescin in the 25-OH 
D3 product. FDA found the purification process to be 
effective based on the analytical work submitted. The analytical method 
for the detection of the 2', 4', 5', 7'-tetraiodofluorescin has been 
appropriately validated by the petitioner and FDA found it to be 
accurate, precise, and acceptable for its purpose.

IV. Conclusion

    FDA has determined that the petition provides information to 
support a determination that the use of 25-OH D3 is GRAS as 
a source of vitamin D3 activity for broiler chicken feeds 
and drinking water based upon the existence of an expert consensus that 
25-OH D3 has been shown to be safe based on scientific 
procedures. The determination was based upon published scientific data, 
corroborating unpublished studies, and other data and information. The 
agency has reached the following conclusions: (1) 25-OH D3 
is a suitable source of vitamin D3 activity for broiler 
chickens; (2) 25-OH D3 at levels up to 69 ppb in feed, or 
34.5 ppb in drinking water, is safe to broiler chickens and to the 
people consuming the broiler chickens' edible meat products; and (3) an 
expert panel also concluded that the available information supports a 
GRAS classification for 25-OH D3 when supplied as a source 
of vitamin D activity in broiler feed at the intended level of use of 
69 [micro]g/kg of feed.
    The evaluation was based on the same type and quality of data as 
would be required to obtain FDA approval of 25-OH D3 for use 
as a food additive. In addition, the majority of the data is published, 
and there is a consensus among qualified experts, based on the data, 
that this intended use of the substance is safe. Corroborating evidence 
has shown that 25-OH D3 is a normal metabolite of vitamin 
D3 and the principal circulating form of vitamin 
D3 in mammals and birds. In addition, the petitioner 
provided testimony of a world-renowned expert on vitamin D who stated 
that 25-OH D3 would be a safer dietary supplement than 
vitamin D, since it does not accumulate in the body and thus, would not 
cause toxicity because of accumulation to toxic levels. Therefore, the 
agency is issuing this final rule affirming that 25-OH D3 is 
GRAS as a source of vitamin D3 activity in broiler feeds and 
drinking water when given to broiler chickens at nutritional levels not 
to exceed 69 ppb in feed or 34.5 ppb in drinking water and when used in 
accordance with additional limitations.

V. Environmental Impact

    FDA has determined under 21 CFR 25.32(r) that this action is of the 
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.

VI. Effective Date

    As this rule recognizes an exemption from the food additive 
definition in the Federal Food, Drug, and Cosmetic Act, and from the 
approval requirements applicable to food additives, no delay in 
effective date is required by the Administrative Procedure Act (5 
U.S.C. 553(d)). The rule therefore will be effective immediately (5 
U.S.C. 553(d)(1)).

VII. References

    The following references have been placed on display in the 
Division of Dockets Management (HFA-305), 5630 Fishers Lane, rm. 1061, 
Rockville, MD 20852 and may be seen by interested persons between 9 
a.m. and 4 p.m., Monday through Friday.
    1. Garret, R. H. and C. M. Grisham, ``The Vitamin D Group,'' in 
Biochemistry, 2d ed., Saunders College Publishing, New York, NY, pp. 
604-605, 1999.
    2. Ameenuddin, S., M. L. Sunde, E. M. Cook, World's Poultry Science 
Journal, vol. 41, pp. 52-63, 1985 (available in the petition: vol. 17, 
pp. 4463-4474).
    3. National Research Council, ``Vitamins,'' p. 15, and ``Nutrient 
Requirements of Chickens,'' pp. 19-34, in Nutrient Requirements of 
Poultry, 9th ed., National Academy Press, Washington, DC, 1994.
    4. Ward, N. E., Journal of Applied Poultry Research, vol. 2, pp. 
286-296, 1993 (available in the petition: vol. 19, pp. 5619-5629).
    5. GRASP 2449, Amendment C-0032: ``Safety of 25-OH D3 to 
Consumers,'' (testimony of Dr. Hector De Luca), November 1999.
    6. National Research Council, ``Vitamin D,'' in Vitamin Tolerance 
of Animals, 1st ed., National Academy Press, Washington, DC, pp. 11-22, 
1987.
    7. GRASP 2449, Appendix E: Expert Panel Report, Life Sciences 
Research Office, Federation of American Societies for Experimental 
Biology, Bethesda, MD, 1994.
    8. Quarles, C. L., Safety study using 25-OH D3 in 
broiler chickens, Colorado AM-1-93, Colorado Quality Research Inc., 
Fort Collins, CO, 1993, (available in the petition: vol. 11, pp. 02005-
02493).
    9. Quarles, C. L., Safety study using 25-OH D3 and 
vitamin D3 in broiler chickens, Colorado AM-2-94, Colorado 
Quality Research, Inc., Fort Collins, CO, 1994, (available in the 
petition: vol. 12, pp. 2494-2862).
    10. Morrissey, J. L., et al., Journal of Nutrition, vol. 107, pp. 
1027-10324, 1977 (available in the petition: vol. 19, pp. 5220-5227).
    11. Janssen, W. M. M. A., H. A. J. Versteegh, P. J. W. Van Schagen, 
Archiv fur Geflugelkunde, vol. 45, pp. 194-200, 1981 (available in the 
petition: vol. 18, pp. 5018-5024).
    12. Kaetzel, D. M., Jr. and J. H. Soares, Jr., Journal of 
Nutrition, vol. 109, pp. 1601-1608, 1979 (available in the petition: 
vol. 18, pp. 5033-5040).
    13. Stevens, V. L. and R. Blair, Nutrition Reports International, 
vol. 35, pp. 755-764, 1987 (available in the petition: vol. 19, pp. 
5465-5474).
    14. GRASP 2449, Amendment C-0027: Safety Study Using 25-OH 
D3 in Broiler Chickens (Project No. ISG-98-2). September 
1998 (available in the petition: vol. 31-34).
    15. Dutta, S. N., Clinical review and evaluation of NDA 018312 
(Organon USA, Inc.'s CALDEROL brand of 25-OH D3), 1979.
    16. GRASP 2449, Amendment C-0028: Human Food Safety of the Use of 
25-OH D3 in Broiler Feed, February 1999.
    17. FDA Center for Veterinary Medicine, Guideline for Industry 
3: ``General Principles for Evaluating the Safety of Compounds 
Used in Food-Producing Animals,'' Part 4: ``Guideline for Establishing 
a Safe Concentration,'' 1994.
    18. GRASP 2449, Amendment C-0029: Safety of the Use of Erythrosin 
in the 25-OH D3 Final Product, May 1999.

List of Subjects in 21 CFR Part 584

    Animal feeds, Food additives.

0
Therefore, under the Federal Food, Drug, and Cosmetic Act and under 
authority delegated to the Commissioner of Food and Drugs, and 
redelegated to

[[Page 12564]]

the Center for Veterinary Medicine, 21 CFR part 584 is amended as 
follows:

PART 584-FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE 
IN FEED AND DRINKING WATER OF ANIMALS

0
1. The authority citation for 21 CFR part 584 continues to read as 
follows:

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

0
2. Section 584.725 is added to subpart B to read as follows:


Sec.  584.725  25-Hydroxyvitamin D3.

    (a) Product. 25-Hydroxyvitamin D3 (9,10-secocholesta-
5,7,10(19)-triene-3[beta], 25-diol).
    (b) Conditions of use. This substance is generally recognized as 
safe as a source of vitamin D3 activity in feed or drinking 
water of broiler chickens when used in accordance with the limitations 
in paragraph (c) of this section.
    (c) Limitations. (1) Not to exceed 69 parts per billion (ppb) in 
feed or 34.5 ppb in drinking water. It shall be used in accordance with 
good manufacturing and feeding practices.
    (2) The product must comply with the following specifications:
    (i) Not less than 94.0 percent 25-hydroxyvitamin D3.
    (ii) Not more than 1 percent of any individual sterol.
    (iii) Not more than 5 percent water.
    (iv) Not more than 20 parts per million (ppm) lead.
    (v) Not more than 20 ppm aluminum.
    (vi) Not more than 1.0 percent solvents and non-detectable levels 
of 2', 4', 5', 7'-tetraiodofluorescin.
    (3) Product labeling shall bear the following:
    (i) A statement to indicate that the maximum use level of 25-
hydroxyvitamin D3 must not exceed 69 ppb in feed or 34.5 ppb 
in drinking water.
    (ii) Adequate use directions to ensure that 25-hydroxyvitamin 
D3 (and all premixes) is uniformly blended throughout the 
feed or drinking water.
    (iii) An expiration date on all premix labeling.
    (iv) A statement on all premix labeling (feed and drinking water 
forms) that 25-hydroxyvitamin D3 should not be used 
simultaneously in both feed and water.

    Dated: March 1, 2007.
Stephen F. Sundlof,
Director, Center for Veterinary Medicine.
[FR Doc. E7-4796 Filed 3-15-07; 8:45 am]
BILLING CODE 4160-01-S