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The Toxicologic and Pharmacologic Effects of Hafnium Salt&"
The Toxicologic and Pharmacologic Effects of Hafnium Salt&"
cress, spleen, adrenal, and small intestine. The method of Draize et al.
(1944) was used to study ocular and skin irritation in rabbits and intra-
dermal irritation in guinea pigs. Six rabbits were used in the ocular
studies, each rabbit had one eye exposed to 0.1 ml of 1:lOOO hafnyl
chloride while the other eye received 0.1 ml of a control saline solution
adjusted to the same pH value. Three rabbits had 1 mg of hafnium
chloride placed in one eye for further study of ocular irritation. Rabbit
skin irritation studies employed 6 animals according to the design of
Draize et al. (1944). Five guinea pigs were used in the intradermal
studies, and the concentrations of hafnyl chloride were 1:lO to 1: 10”.
Effects of hafnyl chloride on isolated guinea pig ileal strips bathed in
Locke-Ringer’s solution were studied in a thermostatically regulated
25ml bath using the Trendelenburg method (1917). Studies were also
made on the isolated rabbit ileum in the presenceof 0.5 ng of nicotine or
2.5 ug of acetylcholine. Eighteen cats of both sexes,weighing 3.54.25 kg
were anesthetized with 0.5 ml/kg of Dial-urethane intraperitoneally. A
six-channel Offner Dynagraph with Statham transducers was used to
record carotid arterial pressure, respiration, nictitating membrane con-
traction, ECG Lead II, femoral arterial pressure, and femoral arterial
flow. The latter was obtained with a 25ml Shipley-Wilson flowmeter
(1951). Preganglionic stimulation of the cervical sympathetic fibers and
the contralateral vagus fibers was accomplished with a Grass model S-4
stimulator at 8v/lO sec. Two hours were allowed to elapse prior to
beginning drug administration. Intravenous dosesof the drugs used were:
hafnyl chloride 0.5-10 mg/kg; hafnyl chloride citrate complex OS-
75 m&kg ; epinephrine 5 pg/kg; acetylcholine 5 pg/kg; histamine
0.5 pg/kg; atropine 2 mg/kg; and sodium citrate 36.8-441.6 mg/kg.
The hafnyl compounds were injected at a constant volume of 2 ml per
dose. Inasmuch as the hafnyl chloride solution had a low pH, control
solutions containing HCI at pH 2.33 to 0.8 were also studied. Where
appropriate, the results were analyzed statistically by the Litchfield-
Wilcoxon method (1949) or standard errors were calculated.
RESULTS
Chemical Considerations
When hafnium chloride (HfC14) contacts moisture or is dissolved in
water there is an immediate liberation of HCl and the formation of
hafnyl chloride (HfOCla). This reaction, whether it occurs in the test
tube or in the gastrointestinal tract, results in a marked lowering of the
240 THOMAS J. HALEY ET AL.
pH. Thus the studies reported herein are in essence based upon the
responses to hafnyl chloride rather than the original compound, hafnium
chloride.
Acute Toxicity
The symptoms of acute toxicity were immediate urination and lethargy.
The first deaths occurred within 24 hours, but the peak was not reached
until 48 hours. A few animals did not die until the sixth day of the study
and all animals were unthrifty during this interval. The intraperitoneal
0 CONTROL
360 0 1.0%
no.l%
t 0 0.01%
0 2 4 6 8 IO 12
WEEKS
FIG. 1. Response of male and female CFN rats to the feeding of various levels
of hafnium chloride in the diet for 90 days. Bars signify the standard errors.
TOXICOLOGY OF HAFNIUM SALTS 241
LD50 for 7 days for hafnyl chloride was 112 (93.3-134.4) mg/kg with a
slope of 1.44 (0.97-2.13).
Chronic Toxicity
Throughout the chronic feeding period of 12 weeks, the animals
receiving the hafnium chloride did not appear to differ from the controls.
The growth curves shown in Fig. 1 indicate that, at the dietary levels of
hafnium chloride studied, there was no significant effect on growth.
Furthermore, the chemical produced no significant effect on the hematol-
ogy of the rats (see Table 1 for a comparison between groups and
sexes). The increases in the cells and hemoglobin between the beginning
and the end of the experiment were probably related to the normal
growth pattern of the animals and not to any influence of the chemical,
because the control group did not differ significantly from the medicated
groups. It should also be noted that all the values in Table 1 were
within the ranges given for the rat by Gardner (1947). The data on
the differential counts are not given in Table 1, because there was no
significant difference between the medicated and control groups. Also
the data were equivalent to the normal distribution given by Gardner
(1947). At autopsy the internal organs of all groups of rats appeared
normal and there were no outward signs of damage from ingestion of
hafnium chloride for 12 weeks. Histopathologic examination revealed
that, of all the tissues examined, only the liver showed consistent damage.
This consisted of perinuclear vacuolization of the parenchymal cells and
coarse granularity of the cytoplasm. Both sexes showed this damage in
S/6 animals at the 1% feeding level whereas only 2/6 males and l/6
females showed such liver changes at the 0.1% and 0.01% levels.
Ocular Irritation
Direct application of 0.1 of 1:lOOO hafnyl chloride pH 2.1 to the
eyes of six rabbits produced no evidence of damage to the cornea or iris or
irritation of the conjunctiva at the l- and 24-hour observation periods.
A similar application of diluted HCl of the same pH also did not affect
the eyes. The placement of 1 mg of hafnium chloride in the eyes of
rabbits resulted in an immediate increase in the rate of blinking and a
redness of the palpebral conjunctiva within 1 hour. At 24 hours there
was no evidence of cornea1 damage, iris damage, conjuctival irritation,
chemosis, or increased lacrimal discharge. Apparently, the natural buf-
fering capacity of the lacrimal fluid was sufficient to counteract the
TABLE 1
HLMATOL~CIC EFFECTS OF HAFNIUM CHLORIDES
Control M 6.20 -c .25 10.38 -c .31 15.00 -c .47 11.60 * 1.39 47 2 1.79 52 k 1.02 12.4 k .41 15.2 f .25
5.4s6.95 9.59-11.29 10.2-19.45 7.25-16.05 4&58 47-53 11.6-14.3 14.4-15.9
F 6.51 & .32 8.99 zk .38 13.20 lr .S8 18.20 2 2.02 47 2 1.07 50 ? .80 12.6 ? .25 13.9 ? .24
5.08-8.35 7.67-11.00 7.00-18.60 10.6-30.80 43-54 47-54 11.3-13.6 12.5-15.2
Hafnium M 6.67 f .23 10.79 k .29 11.10 -c- 1.75 12.05 c 1.47 44 ? 1.78 54 c 1.14 11.9 2 .56 16.6 iI .I4
0.01% 5.82-7.45 9.86-12.03 7.05-18.00 7.6&16.00 39-50 52-59 10.5-14.3 16.0-17.0
F 7.75 2 .40 9.27 -+- .48 10.40 -c 1.25 15.50 zk 2.76 49 k 1.49 so f 1.02 13.0 & .30 15.9 k .33
6.50-9.15 7.89-10.95 5.90-15.40 9.45-28.45 44-53 48-54 12.3-13.9 15.3-17.2
Hafnium M 5.57 r?r: .24 10.32 -c .40 8.80 k 1.05 17.60 -r- 2.07 47 k 1.70 53 k 1.05 12.9 f .58 15.4 k .39
os7c 4.56-6.00 8.99-11.30 5.60-I 1.80 11.45-24.20 42-52 50-56 11.6-15.0 13.6-16.1
F 5.81 + .29 9.19 k .30 10.05 & 2.49 17.90 ? 3.96 47 21 .89 50 k .71 12.8 ? .39 14.1 f .28
4.96-6.80 7.99-10.23 7.60-12.25 7.05-24.25 44-50 47-52 11.5-14.3 13.6-15.4
Hafnium M 5.56 21 .28 10.82 -c .44 10.60 -c 1.19 14.95 k 1.24 46 f 1.30 54 k .91 13.0 2 .36 15.8 -I- .30
1 .O% 4.24-6.20 9.13-12.14 6.40-14.35 11.20-18.65 42-50 52-58 11.6-14.0 14.6-16.5
F 6.01 Ir, .21 9.89 k .39 14.85 ?I 1.42 16.70 ? 3.19 48 2 .82 49 k .80 13.4 -I- .26 15.3 c .2s
5.36-6.88 7.92-10.01 11.20-19.65 9.6s32.00 44-50 47-52 12.5-14.2 14.1-16.0
u Values are means plus or minus standard error and ranges at time 0 and at 12 weeks.
TOXICOLOGY OF HAFNIUM SALTS 243
Pharmacologic Effects
Hafnyl chloride at doses of 0.5-2 mg/kg produced no observable
pharmacologic effects in the cat. A transient hypotension of 15-25 mm
Hg occurred in both carotid and femoral blood pressure coupled with a
decreased femoral blood flow after administration of 5 mg/kg. ,4t this
dose the electrocardiogram showed an increase in the height of the
P wave and a notching of the T wave. Complete cardiovascular collapse
occurred at 10 mg/kg and was followed by respiratory paralysis. Prior
to exitus, the respiratory rate was not affected. The terminal electro-
cardiographic changes included a transient increase in the height of
the P wave until it equaled the QRS complex, decreased height of the
P wave coupled with an increased T wave, absence of the P wave, a
high take-off of the T wave, and finally ventricular fibrillation. Within
the dosage range studied, hafnyl chloride did not affect the physiologic
responses to acetylcholine, epinephrine, histamine, or vagal stimulation.
Furthermore, it had no effect on transmission in the superior cervical
ganglion or on contraction of the nictitating membrane. The above
effects of hafnyl chloride could not be modified by atropine, and
the cardiovascular collapse could not be counteracted by epinephrine.
When complexed with citrate, hafnyl chloride produced pharmacologic
effects that appeared to be identical with those observed with the uncom-
plexed chemical, and cardiovascular collapse occurred at a dose of 60
mg/kg. However, evaluation of the action of citrate alone in doses
equivalent to those used in complexing the hafnyl chloride indicated
that the effects observed were due to citrate, not to hafnium. pH was
not a factor in the results obtained with hafnyl chloride, because in-
jection of the same volume, 2 ml/dose, of diluted HCl within the pH
range 2.33-0.8 produced no pharmacologic effects and was not lethal
to the animals.
TOXICOLOGY OF HAFNIUM SALTS 245
DISCUSSION
REFERENCES
DRAIZE, J. H., WOODAFCD, G., and CALVERY, H. 0. (1944). Method for the study of
irritation and toxicity of substances applied topically to the skin and mucous
membranes. J. Phatvnucol. Exptl. Therap. 82, 377-390.
GARDNER, M. V. (1947). The blood picture of normal laboratory animals. A review
of the literature 19361946.1. Fmnklin Inst. 243, 77-86.
HALE, R. W., and CARMICHAEL, R. L. (1961). Less common elements. Economics.
Znd. Eng. Chem. 63, 108-111.
LITCHFIELD, J. T., JR., and WILCOXON, F. (1949). A simplified method of evaluating
dose-effect experiments. J. Pkarmacol. Exptl. Therap. 96, 99-113.
246 THOMAS J. HALEY ET AL.