SUMMARY
EXPERIMENTAL
Appamtus
Perkin-Elmer 303 and 400 S atomic absorption spectrometers were used;
both instruments were equipped with arc source deuterium lamps for back-
ground correction, and the 400 S spectrophotometer had also a tungsten
lamp for corrections in the range 300-800 run.
Solid and liquid samples were atomized in a graphite furnace, the con-
struction of which has been described elsewhere [lo] _ These analyses were
done with the 303 instrument and were based on measuring peak areas.
140
Samples
The major components of bone are organic collagen and inorganic apatite;
water is bound to both substances_
Thermogravimetric analysis [12] of a human femur bone showed a content
of about 28% protein and about 8% water, the total weight loss up to about
1000C being approximately 36%.
The following materials were analyzed:
(1) a reference sample of calcined animal bone (code name A-3/1 (1974))
issued by the International Atomic Energy Agency (IAEA), Vienna, Austria;
(2) a sample of purified animal bone for medical purposes. (The bone was
from Argentine cattle and had been processed by Weiders Farmas&tiske A/S,
Oslo, Norway. A typical analysis of the product showed the following con-
tents (given in %): calcium, 34; phosphorus, i5; magnesium, 0.17; fluorine,
0.05; and silicon, 0.01.);
(3) eight samples of an ivory tusk which were issued [ 13 ] in connection
with a cooperative investigation of the content and distribution of lead in
hard tissue; they were received as small plates measuring about 20 X 8 X 1 mm
and weighing approximately 0.25 g.
The two samples of animal bone and the hydroxyapatite mere ground to
pass a 270-mesh (53~pm) sieve.
Before analysis by the solid sampling technique, the IAEA and Weider
samples were mixed with equal amounts of graphite powder; the samples
of ivory were not mixed with graphite.
The samples were not dried before analysis.
Decompositions
Portions of hydroxyapatite (about 1 g) were dissolved in 5.0 ml of nitric
acid, and the clear solution was diluted to 25 ml.
The IAEA and Weider samples were decomposed, where specified, in
polytetrafluoroethylene-lined bombs. To portions of about 1 g, concentrated
nitric acid (5.0 ml) was added, and the mixture was heated for about 1.5 h at
ca. 150C. The solutions were diluted to 25 ml. Clear solutions were obtained.
Analysis of hydroxyapatite
The contents of cadmium, lead and manganese in the solid standard were
redetermined by two a.a.s. methods, both based on the use of thestandard
addition technique. In one of the methods, known volumes of metal standard
solution were added to weighed amounts of solid hydroxyapatite. In the
other method, metal standard solutions were added to portions of hydroxy-
apatite solution.
The mixtures were atomized in the graphite furnace, and gave the following
averages (previous data [9,11] for the same batch of material are given in
parentheses): cadmium, 0.34 ppm (0.35); lead, 2.5 ppm (2.4); and manganese
0.89 ppm (0.90). The data established in the present analyses were employed
as the standard values.
Procedures
Before the start of the measurements with the graphite furnace, the hollow-
cathode and deuterium lamps were heated for about 15 min. The flow of
argon was adjusted to 6 ml s-l, and l-15 mg of the sample, sample-graphite
or standard-graphite mixture was weighed in a small tantalum scoop and
placed in the middle of the preheated graphite tube by means of a specially
142
constructed adjustable inserting device. The scoop was reweighed, and the
furnace was moved into its preadjusted position.
Before the atomization of lead and manganese, any chloride in the sample
had to be removed (see below). This was done by adding 20 ~1 of nitric
acid (l+Z) to the solid sample or standard in the graphite furnace, and heat-
ing the mixture for 1 min at about 100C. To ensure the complete removal of
chloride, a new portion of acid was added, and the operation was repeated.
Lead was not detected in the nitric acid blanks. In the present work, chloride
was removed only from the IAEA sample.
In an alternative procedure for the removal of chloride prior to the deter-
mination of manganese, 5 ml of the sample solution wzs evaporated to a small
volume with 5 ml of concentrated nitric acid in a platinum dish at about
120C; the mixture was transferred to a lo-ml volumetric flask and diIuted to
the mark with water.
Interference of chloride
During the preliminary analyses, it, was observed that the values for lead and
manganese obtained by solid sampling of the calcined animal bone from IAEA
were persistently lower than the recommended values, and than the data
obtained by analyzing solutions in the graphite furnace. The results for the
Weider sample and hydroxyapatite did not exhibit this discrepancy. A pos-
sible explanation for this disagreement was the presence of an interfering
element, e.g. chlorine.
Solutions of the two samples of bone and the hydroxyapatite standard were
tested for chloride; the only solution giving a precipitate with silver nitrate
solution was that of the IAEA sample. Similarly, a piece of one of the ivory
samples was decomposed, and the solution was tested for chloride with
negative results.
The interfering effect of chloride on lead in analysis by solid sampling was
confirmed by the following experiments. To portions of the solid Weider
sample (in which chloride could not be detected) varying amounts of potas-
sium chloride solution were added. Samples with and without the addition
of potassium chloride solution, and a potassium chloride blank were then
atomized as described above. The integrals obtained per mg of sample decreased
distinctly as the amount of chloride added was increased. The interfering
effect of chloride on lead.was removed by adding nitric acid to a weighed
portion of the solid sample deposited in the graphite furnace, and
evaporating the mixture to dryness; complete removal of chloride was
secured by repeating the operation.
The interfering effect of chloride on manganese may be removed by
evaporations either in the graphite furnace as described for lead, or in a
platinum dish heated on a hot plate.,Interfering effects of chloride on cadmium
were not found during application of the present techniques.
143
TABLE 1
Results for cadmium, manganese and lead in hydroxyapatite and two samples of animal
bone
[Method A (standard addition). Standard solution is added to the sample solution, with
atomization in the graphite furnace.
Method B (standard addition). Standard solution is added to the solid sample, with
atomization in the graphite furnace.
Method C. Standard solution is added to the sample solution, with atomization in the
flame.
Method D. Solid sample is atomized in the graphite furnace with solid hydroxyapatite as
standard.]
Sample Method Cd Mn Pb
X s T
x
(ppm) (lo) (ppm) &) ippm) k)
Hydroxyapatiteb A 0.33 18 0.94 3 2.5 12
B 0.34 18 0.84 5 2.5 12
Animal bone Values
IAEA recommended
by IAEA not given 32 16c 6.8 15
C - - 33 12- -
A - - 32 11 6.9 15
D 0.072 12 - - 6.8 18
Animal bone C - - 5.4 20- -
Weider A - - - - 9.8 14
D 0.036 22 6.0 22 9.7 10
aAverage result with relative standard deviation (So). bThe sample employed as the solid
standard. Relative standard deviation of the mean value.
144
TABLE 2
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