Menadione Nicotinamide Bisulfite Is a Bîoactîve Source of ...
Menadione Nicotinamide Bisulfite Is a Bîoactîve Source of ...
Menadione Nicotinamide Bisulfite Is a Bîoactîve Source of ...
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nutrient Requirements and Interactions<br />
<strong>Menadione</strong> <strong>Nicotinamide</strong> <strong>Bisulfite</strong> <strong>Is</strong> a Bîoactîve<br />
<strong>Source</strong> <strong>of</strong> Vitamin K and Niacin Activity for Chicks1<br />
GEORGE W. ODÃœHO, THAU K. CHUNG2 AND DAVID H. BAKER3<br />
Department <strong>of</strong> Animal Sciences and Division <strong>of</strong> Nutritional Sciences,<br />
University <strong>of</strong> Illinois at Urbana-Champaign, IL 61801<br />
ABSTRACT Young chicks were fed a vitamin Kdeficient<br />
soybean concentrate basal diet containing<br />
graded levels <strong>of</strong> menadione from menadione<br />
nicotinamide bisulfite (MNB) or menadione dimethylpyrimidinol<br />
bisulfite (MPB) to assess prothrombin time<br />
as a function <strong>of</strong> menadione intake. Prothrombin time<br />
decreased linearly as menadione dose increased from 0<br />
to 400 /ig/kg. Multiple linear regression slope-ratio cal<br />
culations indicated that both sources <strong>of</strong> menadione were<br />
<strong>of</strong> equal potency. To assess niacin bioactivity <strong>of</strong> MNB,<br />
graded doses <strong>of</strong> nicotinamide (0 to 5 mg/kg) from MNB<br />
or nicotinamide were added to a niacin-deficient diet<br />
based upon corn, corn gluten meal and vitamin-free<br />
casein. Weight gain increased linearly as a function <strong>of</strong><br />
nicotinamide dose, and multiple linear regression<br />
analysis <strong>of</strong> weight gain as a function <strong>of</strong> supplemental<br />
nicotinamide intake revealed no significant differences in<br />
slope between the two sources <strong>of</strong> nicotinamide. Using<br />
excess doses, MNB was compared with MPB in acute<br />
(single crop intubation) or chronic (fed in the diet for 14<br />
d) toxicity trials. With a single menadione dose <strong>of</strong> 1600<br />
mg/kg body wt, weight gain in the subsequent 14-d<br />
period was reduced by MNB but not by MPB. Mortality<br />
rates <strong>of</strong> 25 and 17% occurred for MPB and MNB,<br />
respectively, at this dose level. Doses lower than 1600<br />
mg/kg body wt caused neither morbidity nor mortality.<br />
When provided in the diet for a 14-d feeding period,<br />
menadione doses <strong>of</strong> 3000 mg/kg diet from MNB<br />
reduced gain, feed intake, gain:feed ratio and blood he<br />
moglobin concentration. <strong>Menadione</strong> doses <strong>of</strong> 6000 mg/<br />
kg diet were required to produce morbidity <strong>of</strong> this type<br />
when MPB was fed. The results suggest that MNB is<br />
fully effective as a source <strong>of</strong> vitamin K and niacin ac<br />
tivity, and only when doses exceed 1000 times the<br />
chick's vitamin K requirement can morbidity or mortality<br />
be demonstrated. J. Nutr. 123: 737-743, 1993.<br />
INDEXING KEY WORDS:<br />
•menadione nicotinamide bisulfite<br />
•niacin •prothrombin time<br />
•uitamin K<br />
chicks<br />
The biological activity <strong>of</strong> menadione salts or com<br />
plexes as sources <strong>of</strong> vitamin K activity has been estab<br />
lished in several chick studies. Frost et al. (1955)<br />
reported that menadione sodium bisulfite complex<br />
(MSEC)4 was three times more effective than mena<br />
dione (2-methyl-l,4-naphthoquinone). Nelson and<br />
Norris (1961) suggested that MSEC and phylloquinone<br />
were equally active on a molar basis, and both<br />
were more than twice as active as menadione.<br />
Griminger (1965) and Dua and Day (1965) compared<br />
MSEC and menadione dimethylpyrimidinol bisulfite<br />
(MPB) and observed that MPB was more active than<br />
MSEC in reducing prothrombin time <strong>of</strong> chicks. The<br />
source <strong>of</strong> menadione evaluated herein, menadione<br />
nicotinamide bisulfite (MNB), has never been<br />
evaluated for its vitamin K (or niacin) activity.<br />
<strong>Nicotinamide</strong> is known to be an effective source <strong>of</strong><br />
bioavailable niacin activity and has been used as such<br />
for many years in both human and animal applica<br />
tions. Studies by Childs et al. (1952) and Baker et al.<br />
(1973) have suggested 20 to 30 mg niacin/kg diet as<br />
the requirement for maximal growth <strong>of</strong> young chicks.<br />
Baker et al. (1976) later reported that nicotinamide<br />
was 23% more bioactive than nicotinic acid for chick<br />
growth, although subsequent studies by Bao-Ji and<br />
Combs (1986) and Ruiz and Harms (1988) were unable<br />
to confirm this observation.<br />
The objectives <strong>of</strong> this investigation were to assess<br />
the vitamin K and niacin bioactivity <strong>of</strong> MNB (Fig. 1)<br />
relative to known and established standards, i.e., MPB<br />
for vitamin K activity and nicotinamide for niacin<br />
activity. Both acute and chronic toxicity studies were<br />
also conducted to establish a safety pr<strong>of</strong>ile for MNB.<br />
'Supported in part by Vanetta U.S.A., Inc., York, PA.<br />
^Current address: 154 Clemenceau Avenue, #02-07 Haw Par<br />
Centre, Singapore 0923.<br />
3To whom correspondence should be addressed at 328 Mumford<br />
Hall, 1301 W. Gregory Dr., Urbana, IL 61801.<br />
4Abbreviations used: MNB, menadione nicotinamide bisulfite;<br />
MPB, menadione dimethylpyrimidinol bisulfite; MSEC, menadione<br />
sodium bisulfite complex.<br />
0022-3166/93 $3.00 ©1993 American Institute <strong>of</strong> Nutrition. Received 14 September 1992. Accepted 5 November 1992.<br />
737<br />
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738 ODUHO ET AL.<br />
FIGURE 1 Chemical structure <strong>of</strong> menadione<br />
nicotinamide bisulfite.<br />
MATERIALS AND METHODS<br />
Animals and diets. All experiments were con<br />
ducted using male chicks from the cross <strong>of</strong> New<br />
Hampshire males and Columbian females (University<br />
<strong>of</strong> Illinois Poultry Farm). On d 8 posthatching, after<br />
food was withheld overnight, chicks were weighed<br />
and allotted to experimental groups so that each<br />
group had a similar average initial weight and weight<br />
distribution. Diets and water were freely available.<br />
Chicks were kept on a 24-h constant light schedule in<br />
heated, thermostatically controlled batteries with<br />
raised wired floors. All procedures were approved by<br />
the University <strong>of</strong> Illinois Committee on Laboratory<br />
Animal Care.<br />
Vitamin K activity. Assay 1 was conducted to<br />
determine the vitamin K (menadione) bioactivity <strong>of</strong><br />
MNB (45.7% menadione, 32% nicotinamide, Vanetta<br />
SPA, Milano, Italy) when compared with MPB (45.5%<br />
menadione, Vanetta SPA). From hatching to d 7<br />
posthatching, chicks were fed a vitamin K-deficient<br />
soybean concentrate diet (Table 1) estimated to<br />
contain no more than 30 /¿gvitamin K/kg. On d 8<br />
posthatching, duplicate groups <strong>of</strong> four male chicks<br />
were assigned to the dietary treatments. Treatments<br />
consisted <strong>of</strong> graded levels (0, 5, 25, 50 and 400 /¿g/kg)<br />
menadione from MPB or MNB, which were added to<br />
the vitamin K-deficient diet (Table 1) at the expense<br />
<strong>of</strong> cornstarch. The NRC (1984) estimate <strong>of</strong> the dietary<br />
vitamin K requirement <strong>of</strong> young chicks is 500 Mg/kg.<br />
On d 22 posthatching, following the 14-d feeding<br />
period, chicks were weighed and their feed intake<br />
recorded, after which 1 mL <strong>of</strong> blood was drawn from<br />
each chick by heart puncture. Blood from all chicks in<br />
each pen was pooled and plasma was prepared.<br />
Plasma prothrombin time was determined by the<br />
method <strong>of</strong> Quick (1957), using sodium citrate as an<br />
anticoagulant and acetone-dehydrated chicken brain<br />
powder as a source <strong>of</strong> thromboplastin, prepared in our<br />
laboratory using methodology described by Griminger<br />
(1962 and 1965).<br />
Ingredient1<br />
TABLE 1<br />
Composition <strong>of</strong> vitamin K and niacin assay diets<br />
Cornstarch to<br />
Dextrose<br />
Corn<br />
Corn gluten meal (60% protein)<br />
Soybean protein concentrate (65%<br />
protein)<br />
Sucrose<br />
Solka floe<br />
Vitamin-K<br />
assay diet2<br />
100.00<br />
35.40<br />
20.00<br />
g/100 g<br />
3.00<br />
Niacin<br />
assay diet3<br />
to 100.00<br />
40.00<br />
20.00<br />
Vitamin-free casein (84% protein) 6.50<br />
Corn oil 4.00 5.00<br />
Mineral premix4 5.37 5.37<br />
Vitamin premix (K or niacin-free)5 0.20 0.20<br />
DL-choline chloride 0.20 0.20<br />
all-rac-a-tocopheryl acetate (20 mg/kg) + +<br />
Ethoxyquin (125 mg/kg) + +<br />
Sulfathiazole 0.20<br />
Amino acid premix6 1.60<br />
DL-Methionine 0.30<br />
1Cornstarch and dextrose obtained from A. E. Staley, Decatur,<br />
IL; corn gluten meal (60% protein) and soybean protein concentrate<br />
(65% protein) from Archer Daniel Midland, Decatur, IL; sucrose<br />
from Holly Sugar, Colorado Springs, CO; solka floe from James<br />
River, Berlin, NH; vitamin-free casein from U.S. Biochemicals,<br />
Cleveland, OH.<br />
Calculated analysis: 23% crude protein, 14.6 MJ/kg<br />
metabolizable energy and
niacin bioactivity <strong>of</strong> MNB when compared with<br />
nicotinamide (U.S. Biochemicals, Cleveland, OH). On<br />
d 8 posthatching, quadruplicate groups <strong>of</strong> four male<br />
chicks were assigned to the following dietary treat<br />
ments: 0, 2.5, 5.0 and 30 mg/kg nicotinamide (the<br />
latter a positive control) or 0, 2.5 and 5.0 mg/kg<br />
nicotinamide from MNB. The NRC (1984) estimate <strong>of</strong><br />
the dietary nicotinic acid requirement <strong>of</strong> young<br />
chicks is 27 mg/kg. On d 22 posthatching, chicks and<br />
feed were weighed, and chick weight gain, feed intake<br />
and feed efficiency were determined. Chicks that had<br />
been fed 0 and 5.0 mg/kg nicotinamide (from both<br />
MNB and nicotinamide) were killed by cervical dislo<br />
cation at d 22 posthatching, and portions (0.25 g) <strong>of</strong><br />
liver tissue were removed and pooled by replicate.<br />
Liver samples were homogenized and heated in a<br />
boiling water bath for 5 min. Hepatic pyridine nucleotides<br />
(NAD + NADH) were then determined by the<br />
enzyme-cycling<br />
(1969).<br />
procedure <strong>of</strong> Nisselbaum and Green<br />
Toxicity assays. Assay 3 was conducted to de<br />
termine the toxic effects <strong>of</strong> an acute oral (crop intu<br />
bation) dose <strong>of</strong> menadione from MPB (as a standard)<br />
or MNB. Preliminary work in our laboratory had es<br />
tablished that doses <strong>of</strong> MNB or MPB providing mena<br />
dione at 500 mg/kg body weight or less caused neither<br />
mortality nor morbidity. Assay 3 was therefore<br />
designed to provide logarithmic doses <strong>of</strong> menadione,<br />
starting at 100 mg/kg and working up to 1600 mg/kg<br />
body weight. Following dosing at d 8 posthatching,<br />
triplicate groups <strong>of</strong> four chicks at each dose level were<br />
fed a 23% protein corn soybean meal starter diet5 for<br />
a 14-d observation period.<br />
Assay 4 was a 14-d chronic toxicity study involving<br />
triplicate groups <strong>of</strong> four chicks at each menadione<br />
dose level (0, 30, 300, 3000 and 6000 mg/kg diet) from<br />
MPB or MNB. Additions were made to the cornsoybean<br />
meal starter diet. At termination <strong>of</strong> the assay<br />
on d 22 posthatching, blood was obtained by heart<br />
puncture after which 1-mL portions from each chick<br />
in a pen were pooled, resulting in three pooled<br />
samples for each <strong>of</strong> the nine treatments. Blood hemo<br />
globin was determined by the method <strong>of</strong> Crosby et al.<br />
(1954).<br />
Statistical analyses. Common-intercept multiple<br />
linear regression was conducted for data in Assays 1<br />
and 2 using the General Linear Model procedure <strong>of</strong><br />
SAS (1985). Data were first fitted to simple linear<br />
regression equations to establish that Y-intercepts<br />
were not different (P > 0.05) after which the multiplelinear<br />
regression model was fitted<br />
menadione biopotency assessment<br />
to the data. For<br />
in Assay 1, prothrombin<br />
time (s) was regressed on supplemental<br />
menadione intake (/¿g); for nicotinamide biopotency,<br />
weight gain (g) was regressed on supplemental<br />
nicotinamide intake (mg). The ratio <strong>of</strong> slopes (MNB:<br />
MPB) gave estimates <strong>of</strong> menadione or nicotinamide<br />
potency for MNB relative to MPB.<br />
MENADIONE NICOTINAMIDE BISULFITE 739<br />
TABLE 2<br />
Prothrombin time <strong>of</strong> chicks fed diets with graded levels <strong>of</strong><br />
menadione from menadione dimethylpyrimidinol bisulfite<br />
(MPB) or menadione nicotinamide bisulfite (MNB) (Assay I)1<br />
menadioneUS/kg052550400ProthrombinMPBs3528252519time3MNB3533272417<br />
'Values are means <strong>of</strong> duplicate groups <strong>of</strong> four male chicks at d<br />
22 posthatching; average initial weight was 76 g.<br />
The basal vitamin K assay diet (Table 1) was estimated to<br />
contain 0.05) and this indicated that MPB and<br />
MNB were equally efficacious in furnishing bioavailable<br />
menadione.<br />
<strong>Nicotinamide</strong> activity. Addition <strong>of</strong> graded doses <strong>of</strong><br />
nicotinamide or MNB below the niacin requirement<br />
yielded linear (P < 0.01) responses in weight gain, feed<br />
5Contained (g/100 g): corn, 52.8; dehulled soybean meal, 37.0;<br />
alfalfa meal, 1.0; menhaden fish meal, 2.0; corn oil, 4.0; dicalcium<br />
phosphate, 2.20; limestone, 1.00; DL-methionine, 0.20; NaCl, 0.40;<br />
trace mineral premix (Mn, Fé,Zn, Cu, I, Se), 0.15; flavomycin<br />
premix (4.4% flavomycin), 0.05; vitamin premix (retinyl acetate,<br />
cholecalciferol, all-rac-a-tocopheryl acetate, menadione sodium bi<br />
sulfite, vitamin B-12, niacin, D-Ca-pantothenate, rib<strong>of</strong>lavin), 0.10;<br />
choline chloride (60% choline), 0.10.<br />
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740 ODUHO ET AL.<br />
intake and gain:feed ratio (Table 3). Responses at 5<br />
mg/kg nicotinamide were far lower (P < 0.01) than<br />
those obtained when 30 mg/kg nicotinamide was sup<br />
plemented. Hepatic NAD + NADH concentrations<br />
were low in chicks fed the basal diet, but they in<br />
creased (P < 0.05) upon adding 5 mg/kg nicotinamide<br />
from either nicotinamide or MNB. Multiple linear<br />
regression <strong>of</strong> weight gain (g) as a function <strong>of</strong><br />
nicotinamide intake (mg) resulted in a good fit (R =<br />
0.98), with the MNB slope being 87% <strong>of</strong> the slope<br />
value for nicotinamide (Fig. 2). Regression coefficients<br />
(i.e., slopes), however, were not significantly different<br />
(P > 0.05) between the two sources <strong>of</strong> nicotinamide.<br />
Acute toxicity <strong>of</strong> menadione nicotinamide bi<br />
sulfite. Single crop intubations <strong>of</strong> MPB or MNB at<br />
menadione doses <strong>of</strong> 800 mg menadione/kg body<br />
weight or lower did not cause mortality, nor was<br />
growth depressed during the subsequent 14-d obser<br />
vation period (Table 4). With a dose <strong>of</strong> 1600 mg/kg<br />
body wt, however, 3 <strong>of</strong> 12 chicks (25%) receiving<br />
MPB and 2 <strong>of</strong> 12 chicks (17%) receiving MNB died in<br />
the first 2 d following dosing. This dose <strong>of</strong> menadione<br />
from MNB but not MPB caused a depression in<br />
weight gain, feed intake and gain:feed ratio during the<br />
14-d post-dosing period.<br />
Chronic toxicity <strong>of</strong> menadione nicotinamide bi<br />
sulfite. Pharmacologie doses <strong>of</strong> MNB or MNB<br />
provided in the diet at levels up to 6000 mg/kg mena<br />
dione resulted in no mortality during the 14-d feeding<br />
period. At 6000 mg/kg menadione, both sources <strong>of</strong><br />
menadione depressed (P < 0.05) gain, feed intake and<br />
gain:feed, but only MNB depressed blood hemoglobin<br />
concentration (Table 5). At 3000 mg/kg menadione,<br />
MPB produced no morbidity, but MNB depressed all<br />
measures <strong>of</strong> chick performance.<br />
DISCUSSION<br />
<strong>Menadione</strong> nicotinamide bisulfite<br />
is an organic salt <strong>of</strong> two vitamins, menadione bi<br />
sulfite and nicotinamide (Fig. 1). According to the<br />
manufacturer (Vanetta SPA), MNB, a pale yellow<br />
powder, has a melting point <strong>of</strong> 182°C,a pH range <strong>of</strong> 1<br />
to 3.5 and a water solubility (25°C)<strong>of</strong> 19 g/L.<br />
The one-stage prothrombin time chick bioassay<br />
(Quick 1957) is considered the most sensitive assay<br />
for biopotency <strong>of</strong> menadione compounds (Augustine<br />
1985, Griminger 1965, Kindberg and Suttie 1989). In<br />
our hands the prothrombin time assay worked far<br />
better (i.e., better linearity) with thromboplastin iso<br />
lated in our laboratory from chicken brain than was<br />
the case with purchased rabbit thromboplastin. This<br />
agrees with results reported by Griminger (1962).<br />
Griminger (1965) and Griminger and Donis (1960)<br />
used a vitamin K-assay diet similar to that used<br />
herein, and their 21-d-old female chicks appeared to<br />
be more deficient in vitamin K than was the case with<br />
0.5<br />
Supplemental<strong>Nicotinamide</strong>Intake (mg/14d)<br />
FIGURE 2 Multiple linear regression <strong>of</strong> weight gain (Y in<br />
g) on supplemental nicotinamide intake [X in mg) from<br />
nicotinamide (X, *) or MNB (X2, +): Y = 79.3 + 72.7 (±5.3)X\<br />
+ 63.2 (±5.7)Xi, R = 0.98.<br />
our unsupplemented male chicks. They used sulfaquinoxaline<br />
(2 g/kg) to minimize gut synthesis <strong>of</strong><br />
vitamin K. We used 2 g/kg sulfathiazole in our<br />
vitamin K-deficient diet, and this sulfa drug may have<br />
been less effective than sulfaquinoxaline in<br />
minimizing gut synthesis <strong>of</strong> the vitamin. In<br />
Griminger's (1965) work, MSBC was 74% as effective<br />
as MPB in furnishing bioavailable menadione. In our<br />
study (Table 4), MPB and MNB were equally effective<br />
in furnishing bioavailable menadione.<br />
Based upon prothrombin times <strong>of</strong> 17 to 19 s at 400<br />
Mg/kg menadione (Table 2), with normal prothrombin<br />
times for chickens assumed to range from 12 to 25 s<br />
(Smith and Russell 1984), it would appear that the<br />
NRC (1984) estimate <strong>of</strong> 500 Mg/kg for a vitamin K<br />
requirement <strong>of</strong> broiler chickens is a reasonable es<br />
timate <strong>of</strong> the true requirement.<br />
Growth bioassays in chicks work well to establish<br />
niacin activity <strong>of</strong> test ingredients (Baker et al. 1973<br />
and 1976, Jacob and Swenseid 1990), but if the test<br />
sources contain tryptophan as well as nicotinic acid<br />
or nicotinamide, it is difficult to separate niacin<br />
responses from tryptophan responses (Baker 1986).<br />
Our niacin assay diet contained corn, corn gluten<br />
meal and casein. We assumed that nicotinic acid<br />
would be only 20% available in corn and corn gluten<br />
meal (Darby et al. 1975, Yen et al. 1977) and that<br />
bioavailable tryptophan would be relatively low in<br />
these ingredients as well (Yen et al. 1971). We were<br />
therefore surprised that the niacin assay diet, when<br />
adequately fortified with nicotinamide, would not re<br />
spond to supplemental tryptophan. Regardless, the<br />
diet responded markedly to nicotinamide addition,<br />
and the fourfold growth response obtained to addition<br />
<strong>of</strong> 30 mg/kg nicotinamide (Table 3) suggested that<br />
lower doses <strong>of</strong> nicotinamide would result in linear<br />
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<strong>Source</strong>0NANANAMNBMNBPooledSupplemental<br />
MENADIONE NICOTINAMIDE BISULFITE 741<br />
TABLE3<br />
Performance <strong>of</strong> chicks fed diets with graded levels <strong>of</strong> nicotinamide (Assay 2)1<br />
nicotinamideLevelmg/kg02.55.030.02.55.0SEMWeight<br />
gain2731361732641081625Feed<br />
intake2g/14<br />
NAD NADH3nmol/g +<br />
d1682272753642112575Gain:feed2g/kg43659962972551263016Hepatic liver140—215——24517<br />
'Values are means <strong>of</strong> four replicate pens <strong>of</strong> four male chicks during the period 8 to 22 d posthatching; average initial weight was 75 g. For<br />
hepatic NAD + NADH, only chicks fed the basal niacin assay diet and those with 5 mg/kg nicotinamide |NA) from NA and menadione<br />
nicotinamide bisulfite (MNB) were sampled at d-22 posthatching.<br />
2Linear {P < 0.01) response to NA and MNB; 30 mg/kg NA greater (P < 0.01) than all levels lower.<br />
35 mg/kg NA or MNB greater [P < 0.05) than the basal diet.<br />
responses. This was the case (Fig. 2) and the response<br />
to nicotinamide from MNB was not significantly<br />
different from the response to crystalline<br />
nicotinamide.<br />
The LD-50 (dose causing 50% mortality) <strong>of</strong> oral<br />
menadione has been reported to be 500 mg/kg body<br />
weight in mice (Molitor and Robinson 1940) and 804<br />
mg/kg in chicks (Ansbacher et al. 1942). An oral dose<br />
level <strong>of</strong> 750 mg/kg body weight was nonlethal for<br />
chicks. In our acute toxicity study, oral menadione at<br />
800 mg/kg body weight was nonlethal when mena<br />
dione was provided as either MNB or MPB; even at<br />
1600 mg/kg menadione, mortality was 25% or less<br />
(Table 4).<br />
Chronic toxicity studies involving menadione ap<br />
parently have not been conducted in avians (NRC<br />
TABLE 4<br />
1987). Daily oral doses <strong>of</strong> 350 mg/kg body weight as<br />
menadione over a 30-d period have been observed to<br />
depress hemoglobin and blood erythrocyte count in<br />
young rats (Molitor and Robinson 1940). Daily oral<br />
doses <strong>of</strong> 500 mg/kg body weight were lethal to rats<br />
(Molitor and Robinson 1940), while in this study a<br />
dose level <strong>of</strong> 6000 mg/kg diet over 14 d (equivalent to<br />
daily doses <strong>of</strong> approximately 1000 mg/kg body<br />
weight) were nonlethal to young chicks. Avians thus<br />
appear to be more tolerant than mammals to either<br />
acute or chronic pharmacologie dose levels <strong>of</strong> mena<br />
dione.<br />
Although not apparent in mortality data, high<br />
levels <strong>of</strong> MNB were not as well tolerated as high<br />
levels <strong>of</strong> MPB in terms <strong>of</strong> morbidity criteria (Tables 4<br />
Acute toxicity study: performance <strong>of</strong> chicks fed a corn-soybean meal diet with an acute oral dose <strong>of</strong> menadione dimethylpyrimidinol<br />
bisulfite (MPB) or menadione nicotinamide bisulfite (MNB) (Assay 3/*<br />
<strong>Menadione</strong><br />
dose2mg/kg<br />
wt0100200400800 body<br />
d247266244240<br />
intakeMPB395381396403<br />
MNB663656654667<br />
MNB%00000<br />
16003Pooled<br />
211Feed 403MNB3903914163763813579Gain:feedMPB<br />
615g/kg63819632639649630<br />
591MortalityMPB 250000 17<br />
SEMWeightMPB2492622502592692489gainMNBg/14<br />
'Data represent means <strong>of</strong> three pens <strong>of</strong> four chicks during the period 8 to 22 d posthatching; average initial weight was 65 g.<br />
2A single crop intubation was given on d 8 posthatching.<br />
31600 mg/kg body wt <strong>of</strong> menadione from MNB depressed (P < 0.05) weight gain, feed intake and gain:feed.<br />
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742 ODUHO ET AL.<br />
TABLE 5<br />
Chronic toxicity study: performance <strong>of</strong> chicks fed corn-soybean meal diets containing high levels <strong>of</strong> menadione from menadione<br />
dimethylpyrimidinol bisulfate (MPB) or menadione nicotinamide bisulfite (MNB) (Assay 4)1<br />
<strong>Menadione</strong><br />
dose2mg/kg030300<br />
30002<br />
60003Pooled<br />
SEMWeight<br />
gainMPB240239234<br />
d244241240<br />
219<br />
218MNBg/14 1984Feed<br />
intakeMPB358361<br />
351<br />
337<br />
667647MNBS/kg680680690 649<br />
71.6 58.9<br />
337MNB359354348 3136Gain:feedMPB670662 6338HemoglobinMPB78.287.5 72.4MNBg/L86.983.269.0<br />
55.87.2<br />
'Values are means <strong>of</strong> three pens <strong>of</strong> four chicks during the period 8 to 22 d posthatching; average initial weight was 73 g.<br />
2MNB depressed (P < 0.05) weight gain, feed intake, gain:feed and hemoglobin.<br />
3Both MPB and MNB depressed [P < 0.05) gain, feed intake and gain:feed, but only MNB depressed (P < 0.05) hemoglobin. Gain and<br />
hemoglobin depressions were greater {P < 0.05) for MNB than for MPB.<br />
and 5). In an 8-d chronic toxicity study comparing<br />
nicotinic acid and nicotinamide in chicks, Baker et al.<br />
(1976) observed only a slight growth depression from<br />
5000 mg/kg dietary nicotinamide, while 10,000 mg/<br />
kg was markedly growth depressing (20,000 mg/kg<br />
dietary nicotinic acid did not depress growth). It<br />
seems probable that the greater morbidity caused by<br />
MNB compared with MPB results from the combined<br />
effects <strong>of</strong> menadione and nicotinamide in MNB.<br />
The results <strong>of</strong> this investigation suggest that MNB<br />
is fully active as a source <strong>of</strong> bioavailable vitamin K<br />
and niacin activity. If used primarily as a source <strong>of</strong><br />
vitamin K activity, it is worthy <strong>of</strong> note that meeting<br />
the vitamin K requirement from MNB would provide<br />
only a small portion <strong>of</strong> the niacin requirement. Tox<br />
icity studies revealed that MNB has a good safety<br />
pr<strong>of</strong>ile in that dietary levels <strong>of</strong> menadione well over<br />
1000 times the chicks' minimal vitamin K re<br />
quirement were required to cause morbidity. Because<br />
menadione compounds may lose potency when stored<br />
in vitamin or vitamin-trace mineral premixes<br />
(Charles and Huston 1972) stability studies need to be<br />
done comparing MNB to MPB.<br />
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