Electron Paramagnetic Resonance

Biodosimetry in Teeth and Fingernails

A. Romanyukha1,2, R.A. Reyes2, F.
Trompier3, L.A. Benevides1, H.M. Swartz4
Naval Dosimetry Center, 8901 Wisconsin Ave., Bethesda, MD, 20889, USA,
2
Uniformed Services University, 4301 Jones Bridge Rd., Bethesda, MD, 20814, USA,
3
Institut de Radioprotection et de Sret Nuclaire, Fontenay-aux-roses, France,
4
Dartmouth Medical School, Hanover, NH, 03755, USA
1

Outline

EPR dosimetry basics

In vitro X and Q dosimetry in tooth enamel
In vivo tooth L-band dosimetry
EPR dosimetry in fingernails
Conclusions

What is Electron Paramagnetic

Resonance (EPR) ?
Non-destructive magnetic resonance technique
used to detect and quantify unpaired electrons.
Absorption of ionizing radiation generates
unpaired electrons (i.e., paramagnetic centers).
The concentration of radiation-induced
paramagnetic centers is proportional to the
absorbed dose.

EPR: Fundamentals and Principles

There is a net absorption
of energy from the
microwave field at
resonance because of a
greater population of
electrons are in the lower
energy state.
The process is nondestructive because the
population difference
reestablishes itself after
the microwave field is
turned off.
Thus, the history of
radiation exposure is not
destroyed by EPR
measurements.

Optical Imaging

Electron Resonance

Typical frequencies and wavelengths required

for resonance of a free electron in EPR
measurements
Mw Frequency,
Band GHz

Magnetic Sample size

field, T

1.5

0.05

Small animals, whole

human teeth, fingers
in situ

3.2

0.11

Whole teeth, fingers

9.5

0.33

30 - 1000 mg (solid)

20

0.70

10 30 mg (solid)

35

1.22

2 10 mg (solid)

95

3.30

0.25 1 mg (solid)

EPR dosimeters for partial body

exposure
Radiation-induced
radicals are stable only in
hard tissues: teeth, bone,
fingernails and hairs.
Depending on mw band
EPR can be measured in
vivo or in vitro using
specially prepared
samples from human
hard tissues
Finger- and toenails

Finger- and toenails

Characteristics of EPR dosimetry

Non-invasive
Based on a physical process
Not affected by biological processes such as stress
Not affected by simultaneous damage that is likely to occur with
irradiation such as wounds & burns
Applicable to individuals
Measurements can be made at any interval after irradiation up
to at least 2 weeks (fingernails) or indefinately (teeth)
Can provide output immediately after the measurement
Unaffected by dose rate
Can operate in a variety of environments
Systems can be developed so that they can be operated by
minimally trained individuals

In vitro measurements in tooth

enamel samples (X and Q-bands)

Extracted teeth can be

available for in vitro EPR measurements

Validation and Standardization

Four successful International Dose Intercomparisons with
totally more than 20 participating labs
ICRU, 2002. Retrospective Assessment of Exposures to
Ionizing Radiation. Report 68 (Bethesda, MD: ICRU).
IAEA, 2002. Use of electron paramagnetic resonance
dosimetry with tooth enamel for retrospective dose
assessment. International Atomic Energy Agency, Vienna,
IAEA-TECDOC-1331.
EPR dosimetry with teeth is the only method which can
reconstruct external gamma radiation doses (<100 mGy)
individually.

Steps of the method

Tooth collections
Tooth enamel sample preparation
EPR measurements of radiation response
Calibration of EPR radiation response

EPR Biodosimetry
(Teeth)

EPR Biodosimetry
(Teeth)

Hydroxyapatite constitutes:
~95% by weight of tooth enamel
70-75% of dentin
60-70% of compact bones

Romanyukha, et. al, Appl. Radiat. Isot. (2000) and IAEA-TECDOC-1331

EPR Biodosimetry
(Dose Calibration)

EPR Biodosimetry Applications

(Epidemiological Investigations Using Tooth EPR)
Description of
group
Survivors of abombing of
Hiroshima, Japan

Year of
overexposure

Number of
reconstructed
doses

Values of
reconstructed
doses, Gy

Reference

1945

100

0.3-4.0 Gy

Nakamura et al., Int. J. Radiat. Biol. 73,

619-627 (1998)

Mayak nuclear
workers, Russia

1948-1961

~100

0.2-6.0 Gy

Wieser et al., Radiat. Env. Biophys. 2006

Romanyukha et al., Health Phys. 78, 1520 (2000)

Techa riverside
population

1948-1958

~100

0.1-10 Gy

Romanyukha et al., Health Phys., 81,

554-566 (2001)
Romanyukha et al., Radiat. Environ.
Biophys., 35, 305-310 (1996)

Eye-witnesses of
Totskoye nuclear
test, Russia

1954

10

0.1-0.4 Gy

Romanyukha et al., Radiat. Prot. Dosim.,

86, 53-58 (1999).

Chernobyl clean up
workers, Ukraine

1986

660

0 - 2.0 Gy

Chumak et al., Radiat. Prot. Dos. 77, 9195 (1998)

Population of areas
contaminated by
Chernobyl fallout,
Russia

1986

2500

~ 0.1 Gy

Stepanenko et al., Radiat. Prot. Dos. 77,

101-106 (1998)

Semipalatinsk
population

1950s

32

0.3-4.0 Gy

Romanyukha et al., Appl. Mag. Res.., 22,

347-356 (2002)

Conclusion
EPR X-band (9 GHz) dosimetry in tooth
enamel works excellent (LLD<100 mGy,
time after exposure when dose
measurements are possible from 0.01 hr
to 106 yr.
But it requires to have extracted or
exfoliated teeth available for preparation of
tooth enamel

Alternatives to
exfoliated/extracted teeth
L-band (1.2 GHz) non-Invasive in vivo measurements

Q-band (35 GHz) measurements in enamel biopsy

samples (~2 mg) with followed up tooth restoration

Q-band (35 GHz) measurements in

enamel biopsy samples (~2 mg)
with followed up tooth restoration

Description of Q-band
feasibility test
Tooth enamel powder samples for test: 0;
0.1 Gy; 0.5 Gy; 1 Gy; 3 Gy; 5 Gy
Each sample was recorded 3 times in X (100
mg) and Q bands (2, 4 mg)

Recent publication
Romanyukha A. et al. Q-band EPR biodosimetry in tooth enamel
microprobes: Feasibility test and comparison with X band. Health
Physics. 93, 631-635, (2007).

X-band spectrum vs Q-band spectrum

X-band (100 mg), 0.1 Gy

Q-band, (4 mg) 0.1 Gy

0.08

0.01

0.04

EPR signal, a.u.

EPR signal. a.u.

0.06

0.02
0.00
-0.02

0.00

-0.01

-0.02

-0.04

-0.03

-0.06
-0.08
3490

3500

3510

3520

Magnetic field, G

3530

3540

-0.04
12060

12080

12100

12120

12140

12160

12180

12200

Magnetic field, G

1. Q-band has significantly lesser amount of the sample required for dose
measurements
2. Q-band has significantly better spectral resolution of dose response

12220

Dose dependence: X vs Q

Q-band, 4 mg sample

0.10

1.0

EPR radiation response, a.u.

EPR radiation response, a.u.

0.12

X-band, 100 mg sample

1.2

0.8
0.6
0.4

0.2
0.0

0.08

0.06

0.04

0.02

0.00

Radiation dose, Gy

Radiation Dose, Gy

Dental Biopsy Technique

With the enamel biopsy technique a
small enamel chip is removed from a
tooth crown with minimal damage to
Whole Tooth
the structural integrity of the tooth.
A high-speed compressed-air driven
dental hand piece is used with
appropriate dental burs for this
purpose.
Biopsy
Standard techniques for tooth
restoration using light-cured
composite resins rapidly restore the
small enamel defect in the biopsied
enamel surface of the crown.
Preliminary study on discarded teeth
have demonstrated the feasibility of
removing 2 mg enamel chips, the
desired size for sufficient sensitivity In collaboration with B. Pass, P. Misra,
T. De (Howard University)
with Q-band EPR dosimetry.

Q-band biopsy experiment

Tooth enamel biopsy sample 2.2 mg was
irradiated 4 times to the same dose - 4.3 Gy
After each irradiation angle dependence (12
positions) of biopsy sample was studied
Using average, maximum, minimum and median
values of EPR radiation response at each dose
(e.g. 4.3, 8.6, 12.9 and 17.1 Gy) and linear back
extrapolation attempt to reconstruct dose of 4.3
Gy was made

Angle dependence of radiation

response
0.40

Dose = 8.6 Gy

EPR peak-to-peak ampl., a.u.

0.35

0.30

0.25

0.20

0.15
0

50

100

150

200

Angle, degree

250

300

350

Possible approaches:
1. Use average value of radiation
response at each dose;
2. Use maximum value of radiation
response at each dose;
3. Use minimum value of radiation
response at each dose;
4. Use median value of radiation
response at each dose.

Spectra in biopsy sample at different

doses and dose dependences
0.4

4.3 Gy
8.6 Gy
12.9 Gy
17.1 Gy

0.50
0.45

Radiation response, a.u.

EPR dose signal, a.u.

0.3

0.2

0.1

0.0

-0.1

Average
Maximum
Minimum
Median

0.55

0.40
0.35
0.30
0.25
0.20
0.15

-0.2

0.10
12100

12150

12200

Magnetic field, G

Appearance of tooth enamel

spectrum (maximum) of the same
biopsy sample 2.2 mg at different
doses

-2

10

12

14

Dose, Gy

Dose dependences for average, maximum,

minimum and median values of radiation
response at each dose

Results of attempt to reconstruct 4.3 Gy in

biopsy sample (2.2 mg) using different
approaches
Approach

Result of linear
back extrapolation

Average values

5.5 0.8 Gy

Maximum values 7.3 3.6 Gy

Minimum values 5.4 0.7 Gy
Median values
5.4 1.4 Gy

Preliminary conclusions
Tooth enamel biopsy spectra have slightly different shape from
powder spectra, they are more narrow and have higher signal-tonoise ratio for the same dose than powder spectra. However
existence of angle dependence for biopsy spectra makes difficult
dose reconstruction. Possible solution is to use average, maximum,
minimum or median values for each dose for dose reconstruction
Use of average and minimum EPR radiation response values gives
the best results to reconstruct 4.3 Gy, e.g. 5.5 0.8 Gy and 5.4 0.7
Gy, respectively
A possible reason for some dose offset (~1 Gy) is a slope of a base
line of the spectra for this sample
A possible solution is to apply base line correction to spectra before
measurements of peak-to-peak amplitude of radiation response

L-band in vivo

Recent publications

Swartz H.M. et al. Measurements of clinically

significant doses of ionizing radiation using
non-invasive in vivo EPR spectroscopy of teeth
in situ. Appl. Radiat. Isot. 62, 293-299 (2005)
Swartz H.M. et al. In Vivo EPR Dosimetry to
Quantify Exposures to Clinically Significant
Doses of Ionizing Radiation. Radiat. Prot.
Dosim. 120, 163-170 (2006).
Swartz H.M. et al. In Vivo EPR for Dosimetry.
Radiat. Meas. 42, 1075-1084, (2007).

 L-band (1 GHz) of microwaves is better for

realization of in vivo EPR than standard Xband (9 GHz) because it has
Greater tolerance for the presence of water
Relatively large sample volume sufficient for
whole tooth.

Components of in vivo EPR spectrometer

Resonators that will probe teeth in vivo

Magnet system that can comfortably and effectively
encompass the human head
Software for EPR dose response determination
Dose calibration for in vivo L-band measurements

Clinical EPR Spectrometers

Retrospective Radiation Dosimetry

In Vivo EPR Radiation Dosimetry

Under practical conditions with an
irradiated tooth in the mouth of a
volunteer, the dose dependent signal
amplitude is clearly observed. (Acq.
time = 4.5 minutes/spectrum)

EPR Dose Response

1.5

Slope=0.077, SEP=2.41Gy
Slope=0.073, SEP=1.19Gy
Slope=0.081, SEP=2.67Gy

C#27 #11

P-value difference = 0.4

C#11
C#21

0.5

1.0

#22

#27

Patient V107
Patient V110

C#22
0.0

Average P2P RIS EPR signal in canine teeth

2.0

Dose-response relationship
for two head-and-neck radiation patients

10

15
Radiation dose given, Gy

20

25

30

0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0.0

Averaged over 3 days tooth-size adjusted P2P

SE dose prediction = 46 cGy

10

15

30

Radiation given, Gy

Dose-dependence for 6 in vivo teeth, with each tooth irradiated to a different dose and
measured on 3 separate days. Linear regression analysis shows that the standard error
of dose prediction is 46 cGy.

EPR biodosimetry in tooth enamel for

partial body dose assessment
X-band EPR is ready to use for forensic dose
assessment. Could be carried out on compact
and transportable (< 150 kg) EPR spectrometer.
Dose level <100 mGy.
Q-band biopsy potentially is able to measure
doses < 500 mGy in biopsy tooth enamel
samples 2-4 mg.
L-band in vivo EPR potentially is able to
measure doses as low as 3 Gy. Needs some
additional development.

Finger-and toenails facts

The major component of

fingernails is a -keratin. This
protein is built up from three,
long -helical peptide chains
that are twisted together in a lefthanded coil, strengthened by S
S bridges formed from
adjacent cisteine groups.

Typical available amounts of nail parings

are up to 120 mg for fingernails and up to
160 mg for toe nails
Nails grow all the time, but their rate of
growth slows down with age and poor
circulation
Fingernails grow at an average of onetenth of an inch (3 mm) a month. It takes 6
months for a nail to grow from the root to the
free edge
Toenails grow about 1 mm per month and
take 12-18 months to be completely
replaced
The nails grow faster on your dominant
hand, and they grow more in summer than in
winter

Recent development
Romanyukha A. et al. EPR dosimetry in chemically
treated fingernails. Radiat. Meas. 42, 1110-1113,
(2007).
Trompier F. et al. Protocol for emergency EPR
dosimetry in fingernails. Radiat. Meas. 42, 10851088, (2007).
Reyes R.A. et al. Electron paramagnetic resonance in
human fingernails: the sponge model implication. To
be published in Radiat. Env. Biophys. (2008)

New insights in EPR fingernail

dosimetry
Fingernails can be considered as a sponge-like
tissue which behaves differently from in vivo
fingernails when mechanically-stressed after
clipping
Most of previously published results on EPR
fingernail dosimetry were obtained on stressed
samples and not applicable to life-scenario
situation
Unstressed fingernails have more significantly
stable and sensitive radiation response which
can be measured with EPR

Radiation-induced signal
in unstressed fingernails
1 Gy
5 Gy
8 Gy

0.2

RIS, a.u.

0.1

0.0

-0.1

-0.2

3450

3500

Magnetic field, G

RIS parameters: g=2.0088 H=9 G

3550

3600

RIS spectra obtained by

subtraction of BKS
spectrum recorded prior
irradiation

Result of dose reconstruction in the

sample irradiated to 4 Gy 5 days before
reconstruction
0.50
0.3

Original signal
after treatment
+2 Gy
+4 Gy
+6 Gy
+11 Gy

0.1

0.45

EPR dose response, a.u.

EPR signal, a.u.

0.2

0.0

-0.1

Parameters of the data fit

with Grun model
Imax=0.5513
D0=7.3 Gy
DE=3.66Gy

Grun model

0.40

0.35

0.30

Grun model:
A = Imax(1 - exp(-(D+DE)/D0)),
where A= EPR dose response,
Imax = max EPR dose response (saturation level),
DE=the dose to be determined
D0= characteristic saturation dose

0.25
-0.2
3420

3440

3460

3480

3500

3520

3540

Magnetic field, G

3560

3580

3600

0.20
0

Added dose, Gy

Reconstructed dose 3.66 Gy, reduction

10

12

Variability of dose dependence in

fingernails

Dosimetric properties of
fingernails
Optimal sample mass is 15-20 mg (nailparings from 2-3 fingers)
Measurements time 5 minutes (10 scans)
Achievable lower dose threshold ~ 1 Gy
RIS fading half-time 300 hr (~2 weeks)

Conclusions
Part of EPR
LLD,
body
band/freq Gy

in vivo/
amount

Time
stability

Tooth
enamel

0.1

50 100 mg 106 yr

Tooth
enamel

0.3-0.5 2-4 mg

106 yr

Tooth

3-5

In vivo

106 yr

Fingernails

0.5-1

20-30 mg

~2 wks

Acknowledgements
G. Burke, E. Demidenko, C. Calas, I.
Clairand, T. De, O. Grinberg, A. Iwasaki,
M. Kmiec, L. Kornak, B. LeBlanc, P.
Lesniewski, P. Misra, C. Mitchell, R.J.
Nicolalde, B. Pass, A. Ruuge, D.A.
Schauer, J. Smirniotopoulos, A. Sucheta,
T. Walczak

Disclaimer
The views expressed in this presentation are those of the
author and do not reflect the official policy or position of
the Navy and Marine Corps Public Health Center, Navy
Bureau of Medicine and Surgery, Department of the Navy,
Department of Defense, or the U.S. Government.

www.Biodose-2008.org