Calcif. Tiss. Res.

23, 113-114 (1977) Calcified

Tissue Research
9 by Springer-Verlag 1977

Absorption Spectra of Bone

J. Behari, 1 S.K. Guha, 1 and P.N. A g a r w a l 2

l Biomedical Engineering Division, Indian Institute of Technology and All India Institute of Medical Sciences, New Delhi
2Department of Physics, Indian Institute of Technology, New Delhi-29, India

S u m m a r y . A b s o r p t i o n spectra o f h u m a n bone and its
two m a j o r constituents (collagen and apatite) were ob-
tained in the wavelength region extending from 2000 to
12,000 &. In the last two cases a minimum t r a n s m i s -
sion (maximum absorption) was uniformly observed in
the ultraviolet region. The two samples after exposure
to ultraviolet radiations show a shift in the peak
positions. A b s o r p t i o n peaks in the total bone.spectra are
not reproducible in its two constituents. After exposure
to UV radiations the position of m a x i m u m absorption
is displaced. A possible interpretation o f ' t h e observed
results is:presented.
K e y words: Bone - - A b s o r p t i o n S p e c t r a - - Collagen
-- Hydroxyapatite
Introduction
In an earlier p a p e r the present authors (Behari et al.,
1975) examined the effect of UV' radiations on the
electrical conducti 88 of bone. T h e present w o r k is an
extension of these investigations to include the ab-
sorption spectra o f bone and its two m a j o r constituents
in the form of thin films. The choice o f UV light was
dictated b y the fact that it will produce certain mole-
cular dissociation which will be reflected in the ab-
sorption spectra.
adherent from the surface. The samples were then left at room
humidity for two days. For absorption studies, the sample was
placed in the sample holder of the spectrophotometer with the direc-
tion of beam propagation transverse to the long axis of the sample.
The spectra were obtained in the range 2000-12,000 •. The
samples were then exposed to UV lamp (3500 &, 250 watt) for 2 h
at a distance of about 150 ram. The spectra of irradiated sample
were examined under identical conditions.. The experiment was
repeated for a few samples of each (bone, collagen, and apatite).
Results and DiScussions
Figures 1-3 s h o w the absorption spectra o f bone and
its two major constituents, collagen a n d apatite. The
importance Qf'hydrogen bonds has been emphasized in
both constituents ( R a m a c h a n d r a n , 1967; Hamilton,
1968). Since bone is a hydrogen bonded solid, the two
constituents a f bone should show similar absorption
spectra. In the unexposed collagen and apatite mineral
there is an absorption m a x i m a (minimum trans-
mittance) near 2600 A and 3000 A respectively, agree-
100 Bone collagen
80
Unexposed
E 60
.O

u] Exposed to UV light
/"
/
Materials and Methods 40 /
J
J

The bone specimen was obtained from fresh and mature human 20
tibia and ground with coarse and fine powder to obtain specimens
of the order of about 50/~ thick. Apatite and collagen samples were
obtained from the total bone by the methods of Becker (1965). The P
sample surfaces were treated by ultrasonic washing to remove any 2000 4000 8000 8000 10000 12000
.~., ( A~ )
Send offprint requests to J. Behari at the above address Fig. 1. Transmittance spectra of bone
114 J. Behari et al.: Absorption Spectra of Bone

Bone Unexposed Bone apotile

100 100 -

/ \ ! Unexposed
80 / \,,] /
/
/
/
80-
t t, /
t

ilZ,
/
g 60 / 60r-
/ g
/
/
co
/
E /
~40 40-
1:3
k\ U
Exposed to UV light Exposed to UV light
k\
20 20- j l

L
I i L i I I I I "v',,293c7-~ I E i
0 2000 L O 0 0 6000 8000 10000 12000 0 2000 4000 6000 8000 10 000 12000
~(A") ,Z(A~)
Fig. 2. Transmittance spectra of collagen Fig. 3. Transmittance spectra of apatite mineral

ing well with the reported values of activation energy in be due to absorption taking place in the separated
such solids. Strong absorption bonds in the 2 0 0 0 - 4 0 0 0 structure. Another peak lies in the infrared region, and
A region are correlated with molecular structures con- our results in UV and near UV region are in good
taining chains or rings of conjugated double bonds qualitative agreement with fluorescence spectra results
(Sinsheimer, 1955). Strong absorption bands were pre- on bone (Becker, 1964).
sent in the near UV region, in agreement with the mole-
cular structure o f bone components. On exposure to Acknowledgements. Work partially supported by Family Planning
UV light, one peak remains almost at the same position Foundation, New Delhi, India.
(in both cases) and, in addition, one peak is displaced
towards the higher wavelength. This suggests that on
exposure to UV radiation, some additional excited levels References
are created which reduce the energy of electronic
excitation and the absorption peak thus shifts towards Becket, R.O., Brown, F.M.: Photoelectric effects in human bone.
the long wavelength region. Nature 206, 1325-1328 (1965)
Behari, J., Guha, S.K., Agarwall, P.N.: The effect of ultraviolet
In total bone, there is an absorption peak at 4950
radiation on the electrical conductivity of human bone. Calcif.
A, in agreement with the results of Becker et al. (1964) Tiss. Res. 19, 223-227 (1975)
near 5000 A. A p a r t from this, an absorption peak also Glimcher, M.J.: Molecular biology of mineralized tissue with par-
occurs in the vicinity of 5700 A. However, the ab- ticular reference to bone. Rev. Mod. Phys. 31,359-393 (1959)
sorption peaks observed in collagen and apatite mineral Hamilton, W.C.: Structural chemistry and molecular biology (A.
are not reproducible in whole bone. In bone, the apatite Rich and N. Davidson, eds.). San Francisco: W.H. Freeman
and Company 1968
and collagen are probably cemented together by poly- Ramachandran, G.N.: Treatise on collagen (G. N. Ramachandran,
saccharides (Glimcher, 1959). Binding through poly- ed.), Vol. 1, pp. 103-179. London: Academic Press 1967
saccharides would be by Van der Waals virus, or by Sinsheimer, R.L,: Radiation biology (A. Hollaender, ed.), Vol. II,
hydrogen bonds, or by both. This bonding seems to be pp. 165-201. New York: McGraw Hill 1955
effected on exposure to UV light, probably by the
breakage of certain bonds. The peak near 4100 A may Received January 1 / Accepted October6, 1975