Copyright iD IFAC Control Applications in Post-Harvest and

Processing Technology, Budapest, Hungary, 1998

MEASUREMENT OF FREEZING PROCESS OF AGRICULTURAL

PRODUCTS BY IMPEDANCE SPECTROSCOPY

Kiyohiko Toyoda and Roumiana Tsenkova

Agricultural and Food Process Engineering Laboratory

Department ofAgricultural and Environmental Engineering
Faculty ofAgriculture, Kobe University, Kobe JAPAN

Abstract: Analysis by bioelectrical impedance spectroscopy was conducted to pursue the

behavior of cell tissues in apple fruits during freezing. The relationships between the
parameters in an electrical equivalent model of the cell tissues and the progress of freezing
were investigated. The extra- and intra-cellular resistance of the model showed the well
correlation with a frozen ratio calculated from Plank's equation and these parameters were
found to be useful to detect the beginning and the end of the freezing and the progress of
freezing of the cell tissues of agricultural products. Copyright iD 1998 IFAC

Keywords: Electrical Impedance, Least-square method, Parameter approximation,

Resistance, Capacitance

1. INTRODUCTION measurement can be expected. Furthermore, freezing

The purpose of freezing agricultural products is to
preserve them at low temperature and it is capable to
slow down chemical and enzyme reactions and to
retard the growth of microorganism which causes
quality reduction and spoilage.
During freezing, the behavior of the water in cell
tissues largely influences physical, biological and
chemical changes in cell tissues. For instance,
rupture of a cell wall by ice formation and
denaturation of protein in a cell are caused by the
freezing operation at an improper freezing rate.
Thus, it is important to know the progress of the ice
formation and its rate during freezing. Freezing
process of cell tissues can be measured by the
temperature measurement, however, the temperature
measurement cannot directly give the progress of
freezing.
The electrical properties of agricultural materials are
expressed as a function of temperature, composition
especially moisture content, phase status and so on.
Water in a cell tissue exists as aqueous ionic solution
and the concentration of the dissolved ion in the
solution increases with the progress of the ice
formation during freezing. Therefore, the detection
of the progress of freezing by an electrical
injuries in cell tissues could be detected by an
electrical measurement. (Zhang, et al., 1991a)
Recently, various applications of biological
impedance speetroscopy have been attempted to
investigate characterization of plant tissues, cold
acclimation, heat injury of plant tissues, ripening of
fruit, gelatinization of starch and drying of
agricultural products. (Zhang, et al., 1990; Zhang, et
al., 1992; Zhang, et al., 1993; Harker, et al., 1994;
Toyoda, et al., 1994; Toyoda, et al., 1997)
Common feature in these studies is to analyze the
phenomena occurring in cell tissues on the basis of
their configuration, namely through the behaviors of
the intracellular and extracellular parts and cell
membrane.
Accordingly, a similar analysis in freezing of cell
tissues may provide useful information on the above
various changes in a freezing process of cell tissues.
In this study, the impedance measurement and
analysis were applied to freezing process of
agricultural materials and the followings were
investigated :
i) To investigate the feasibility of the application
of the impedance measurement to the detection of
freezing process
ii) To investigate the relationship between the

89
 freezing progress calculated from Plank's equation
and the values of the parameters of an electrical
equivalent circuit model adopted for explaining the
impedance characteristics.
2. MATERIALS AND METIlODS
2. J Materials and Pretreatment
Fresh apple fruits of Jon-a-Gold variety obtained
locally were used for the experiments and these were
cut to cylinder pieces (14.0 mm in diameter and 50
mm in length) and rinsed with deionized water.
After wiping the water off the surface, tissue pieces
were wrapped with a layer of Parafilm and left in a
refrigerator at least one hour before experimentation.
2.2 Freezing experiments
Immersion freezing and blast freezing experiments
were carried out to evaluate the effect of the heat
transfer within a sample tissue and the convective
heat transfer on a freezing process respectively.
In the immersion freezing experiments, a single
piece of tissue with a film was immersed in
refrigerant of an ethyleneglycol solution.
Refrigerant temperature was controlled at -20 ±O.5
QC and the cooling rate was 90 QC lb. The properties
on heat transfer within the sample were determined
by the temperature measurement with three
replications.
Blast freezing experiments were carried out with two
sample pieces for the measurements of material
temperature and electrical impedance respectively.
Two samples were cooled simultaneously in a cold
air blast controlled at the temperature between -26
and -20 QC and the cooling rate was within a range
between -30 and -60 QC lb.
In both these experiments, temperatures were
measured at the surface and in the center of the
samples and in a refrigerant with T-type
thermocouple (CC) at 5 seconds intelVals. For the
measurement of the center temperature, the
thermocouple was inserted from the cylindrical edge
and the both edges of the sample tissue were
wrapped thicker with layers of the film. Surface
temperature of the sample was measured under the
wrapped film.
In the experiments of immersion freezing, the
impedance measurement was difficult because of the
penetration of the refrigerant solution containing
electrolytes into the sample. Impedance
measurement was carried out only in the blast
freezing experiments.
90
2.3 Analysis ofheat transfer duringjreezing
Many mathematical models including analytical and
numerical methods have been developed. One of the
simpler methods to predict freezing time is to use
Plank's equation. Plank's equation has the
disadvantages derived from the pseudo steady state
assumption, however, the equation can provide for
an intuitive aspect on the effects of the freezing
conditions such as temperature of the refrigerant and
the heat transfer properties on the freezing time.
The freezing time of a cylindrical material is as
follows:
f f·p,{(~)+( ::,)}/Vf-rJ
t =L (1)
tf: freezing time, s
Lf : latent heat, kJ/kg
p' :density of frozen material, kg/m3
2
h : convective heat transfer coefficient, W/(m 'K)
k': thermal conductivity of frozen material,W/(m-K)
R : radius of a cylindrical specimen, m
1j :freezing temperature, QC
Ta :refrigerant temperature, QC
In the immersion freezing experiments, the surface
temperature was found to approach quickly to the
refrigerant temperature and the resistance of the
convective heat transfer at the surface could be
negligible. Therefore, the first term Lf p'R/(2h) in
the right side could be emitted and then, the second
term on the resistance of the heat transfer within a
material, LJflR21(4k') was determined from the
measured data of Tf , Ta and tf.
In blast freezing, both these terms may affect the
freezing time and the sum of both terms in blast
freezing was determined from the measured data in
similar way. The value of the first term in the blast
freezing experiment was obtained by subtracting the
first term of the immersion freezing from the sum
value calculated from the freezing time in the blast
freezing.
The expression of a freezing process derived from
Plank's equation is as follows:
If = t:~~){(~)+(::,)}-t:~~{iJ
.{(~)+(::.}(l-lnt~JIJI (2)
x : radius at the freezing front in the material, m
The radius of freezing front at the given time, t was
calculated by Eq.(2) and the frozen volume ratio, tP
was defined as a ratio of the frozen area to the whole
cross area of the cylindrical samples in this paper as
follows:
tP =J-(xIR/. (3)
 functions as capacitor electrically. Therefore, some
2.4 Electrical impedance measurements changes in concentration of intra- and extra-eellular
electrolytes influence the electrical impedance of cell
The electrical impedance of a sample tissue in the tissues.
blast freezing experiments was measured by 4 •
~ ·A .
inserting four Pt electrodes into the sample in the
~.
direction perpendicular to the long axis of the .. 3 _. .' -
K ~ o·
-....
..,.,i
.~
cylindrical sample. The inner two electrodes were 12 .......

.
10"'"
I'--.. ...ottt.
16 mm apart. The applied voltage was less than 10 .~

mV. The sample tissue was kept in a chamber for

blast freezing with another sample for the
temperature measurement.
ae1 101 •
...,.,
... -' • Jt7 711

LBode-diqram
Impedance spectra consisted of 200 frequency points
at approximately equal intervals between 500 Hz and .~ #'z •
I
50 kHz and the quick measurement at 30 seconds to go :
2 minutes intervals was available by a FFT (fast -
~
t t~
Fourier transform) noise impedance method. The
measurement apparatus was mainly composed of a
arc :I
FFT spectrum analyzer, a pseudo random noise rCiQ)
generator, a potentiostat and a personal computer in
Fig. 1. b. Cole-CoJe. plot

FFf spectrum analyser Fig. 2 Typical Bode diagram and Cole-Cole plot of
cell impedance

2.6 Equivalent Circuit Model

Hayden et al. considered the cell structure into the

development of an electrical equivalent circuit model
and proposed a model called 'Hayden model' in Fig.
3. Hayden model assumed that a cell tissue was
composed of intra- and extra-eellular electrolytes
and a cell membrane. Afterwards Zhang et al.
(199Ib) presented a double shell model including an
personalcomputer explanation of the ~2 dispersion which depends on
the organs' membrane in the cells. However, little
Fig. 1. Configuration of On-line Impedance effect of the ~2 dispersion in the impedance
Measurement System using FFT algorithm measurement at the frequency below 100 kHz was
suggested and the impedance property could be
approximated with Hayden model in Fig. 3.
2.5 Impedance characteristics of biological
materials

A Bode diagram and a Cole-Cole plot as shown in

Fig 2 usually represent impedance property of
biological materials. A typical Bode diagram of cell Fig. 3. Hayden model
tissues shows two decreases in an impedance
amplitude corresponding to ~1 and ~2 dispersions Hayden model was composed of three elements with
which are observed at frequencies from 10kHz to 1 four parameters; apoplasmic (extracellular)
MHz and over 1 MHz respectively. Cole-Cole plot resistance Ra, symplasmic (intracellular) resistance
indicates the relationship between a real part and an Rs, membrane capacitance Cm and phase angle y.
imaginary part of the complex impedance and the ~1 The membrane impedance with a constant phase
and ~2 dispersions were represented as two circular angle y in the model, Zm could be represented in the
arcs in the Cole-Cole plot. following equation:
The ~1 dispersion of the cell tissues depends on the
cell structure. Macroscopically, cell tissues can be cos '11 + j . sin '11
Zm =---'-----'-----'- (4)
regarded as an electrical system in which a cell Cm·O)
membrane isolates intracellular electrolytes from The whole impedance Z of the cell tissues was
extracellular electrolytes and the membrane derived from the cell structure of the model in Fig. 3

91
 Z- ~-~-
1 -~
- (1/ Ra )+ 1/(Rs + Zm) Table 1 Experiment conditions in immersion and
blast freezing
= Z' + Zj . Z" (5)
Complex nonlinear least square method (Macdonald, 1f ('C) Ta ('C) tf (s) terms in Eq. (2)
1987) was applied to determine the model immersion -1.5 -19.7 525 Lf p'R2/(4k,)
parameters (Ra, Rs, Cm and vi) and Levenberg- =1.919x108
freezing
Marquardt algorithm was adopted in the numerical
calculation.
blast -1.9 -21.9 1170 Lf p'R/(2h)
freezing =1.999xI0 6
3. RESULTS AND DISCUSSION
._------------------------
Average value of the term Lf p'R 2/(4k,) in immersion
3.1 Temperature history during air blast freezing
freezing was 1.919 x 108 and it was much larger
than the estimated value from the physical
Typical temperature history during blast freezing
properties. The difference of both values might be
and the frozen ratio calculated from Eq.(2) are
due to the resistance of heat transfer of the layer of
showed in Fig. 4. Initial temperature of a sample
the Parafilm. Subsequently, Lf p'R/(2h) was
tissue was 10 QC and the cooling time was required
calculated in the same way as mentioned in the
to reduce the sample to the freezing point.
analysis. The Biot number was in a range of 0.3 8 to
Consequently, three periods with different behaviors
0.67. These results are showed in Table 1.
of the center temperature could be obselVed
through a whole freezing time: the period I for
cooling from the initial till 300 seconds later at
3.2 Impedance variation during freezing
which the center temperature reached the freezing
point at -1.9 QC, the period 11 for freezing from 300s
A semi circular arc of the measured impedance
till 1170s at the end of the "plateau" of the center
could be obselVed on the Cole-Cole plot. The
temperature and the period rn for further cooling to
parameters of Hayden model were determined by
reduce the center temperature to the temperature of
applying Eq.(5) to the measured data and the
the air blast. .
calculated values by Hayden model could agree well
The calculated frozen ratio approached to unity at
with the measured values at the periods I and 11. At
the end of the plateau of the center temperature -
the period rn, a radius of the circular arc was so
21.~C.
large that little piece of the circular arc could be
In the immersion freezing experiments, the freezing
obselVed within the measured frequency range and
time was determined from a curves of the center
consequently, Hayden model could not sufficiently
temperature of a sample.
adaptable to the impedance at the period Ill.
With a progress of freezing, a radius of the circular
arc on the Cole-Cole plot enlarged gradually and the
arc shifted its position as shown in Fig. 5.
10 .1 The parameters of Hayden model, extracellular
$:>
resistance Ra, intracellular resistance Rs, membrane
capacitance Cm and phase angle If/ were determined
ill and showed the distinctive changes with a progress
of freezing as shown in Fig. 6.
4
xlO
o 1000 2000 3000
time (5)

708 .

1000
o n 5 10 4

1000 2000 3000 time (s) resistance (ohm) x 10

time (5)

Fig. 4. Temperature history during air blast freezing Fig. 5. Variation of the electrical impedance during
and frozen ratio air-blast freezing

92
 lO~.--------..---
........- - - , . . . . . . - - .
extracellular resistance was found. This delay
suggested the ice formation in extracellular part
1170.
might occur prior to the freezing in the intracellular
300s
1 part in the experiments. Intracellular resistance
turned from an increase to a decrease after the
completion of the freezing. .
I o·!-----:::t:=------;=;;-----;;:tin--'-~ Membrane capacitance, Cm shows a decrease till
o 500 10QO 150.0 2000
388 seconds and this behavior was different from the
time (sec)
3)( 10' past observations by the authors in which the
capacitance decreased with a rise in. tempera~e.
The decrease in the membrane capaCitance dunng
the period 11 might be caused by freezing.
Phase angle Ij/ is regarded as an index representing
the homogeneity of the impedance characteristics in
the tissue. An ideal uniform distribution
oOL..---=5O~.O:::-----;I;:::O'="QO=-----;;;15~OO"·-----'~2000 ..
time (sec) corresponds to a phase angle of 90° and the phase
angle decreases with the enlargement of distribution.
Therefore, no significant change in the distribution
was observed during the period I and the
homogeneity of the electrical characteristics within
the cell tissues reduced during the period 11.
The relationship between these parameters and the
frozen ratio t/J was investigated and showed in Fig.
500 10QO ) 1500 2000
time {sec 7. The parameters except a phase angle showed
65.------.-----.-----,.,..,..--, different behaviors before and after the freezing ratio
of 0.5, which corresponds, to the boundary time
between the cooling period and the freezing period.
This problem might be caused by the siml?l~~
$ 75 : · ; ~.H~:.~ .. :..'."-- _.... assumption in Plank's equation that the lru~al
300s 1 . temperature of the material was equal to the free~mg
700.L----=5Od-0~-·,..,.·~I:"i:O':::~O;:-.~...;..·tl15;';;0:;;-0--~2000. point. Therefore, the authors introduced a modified
.·time(sec) assumption that freezing began at the time when the
center temperature reached the freezing point and
Fig. 6. Variation of impedance parameters
eliminated the influences of the cooling time to the
calculation of the frozen ratio.
Extracellular resistance, Ra was 5.51 n at the initial
For this reason, the freezing time in Table 1 were
temperature of 10 °C and increased up to 7.26 n at
replaced by the new values of 390 seconds for the
the end of the period I during cooling. Generally, an
immersion freezing and 941 seconds for the blast
electric resistance of the electrolytic solution rises
freezing. Consequently, the values of L.[p'R]/{4k') an~
with a fall in temperature and the increase in the
Lt p'R/{2h) were calculated as 8.36xlO and 2.lOx10
extracellular resistance from the initial time till 300s
respectively.
may be due to this effect of the temperature.
The relationship between the model parameters and
Subsequently, the resistance continued to increase at
the frozen ratio was determined in same procedures
a slight higher rate during freezing process and
as mentioned before. Figure 8 shows the
reached 14.3 n at the end of the period II. The
relationship between the normalized model
electric resistance increases with a rise in
parameters and the frozen ratio. The normalization
electrolytes concentration in the tissue solution.
for the extracellular resistance were calculated as
During freezing, the electrolytes in cell tissues are
follows:
concentrated by ice formation in the solution.
Therefore, it was suggested that the increases in ~a - Ra at ft=ing po .. )
(6)
extra- and intra-eellular resistance might be due to XR. = ~a at tJw nod _/ ~ - Ra at /Ift=Utg po .. )

the ice formation. Furthermore, The extracellular

For other parameters, the normalization was done
resistance rose more rapidly at the period ill.
similarly. In Fig. 8, the good correlation can be
Consequently, each period can be discriminated
observed between the normalized parameters and the
from the increase tendency of the extracellular
frozen ratio. Especially, the intracellular resistance
resistance Ra.
shows linear correlation with the frozen ratio.
Intracellular resistance, Rs also showed an increase
Accordingly, it was found that the impedance
during the period I and 11. The reasons of these
measurement was applicable to detect the begging
increases were similar to the extracellular resistance.
and the end of freezing and the progress of freezing
In the detail investigation, about 120 seconds delay
in online.
of a rise in the intracellular resistance to that of the

93
 REFERENCES

:~
Harker, F.R, J. H. Maindonald {1994). Ripening of
nectarine fruit, changes in the cell wall,
vacuole, and membranes detected using
..
0.2 0.4 ~ (_ )0.6 0.6 1
electrical impedance measurements, Plant
PhysioJ., 106, pp.l65-171,
Hayden, RI., C.A.Moyse, F.W.Calder , D.P. Craford

~·:3
and. D. S.Fensom (1969). Electrical impedance
studies on potato and alfalfa tissue, Journal of
Experimental Botany, 20 (63), pp. 177-200

0.4 ~ Macdonald, 1.R{1987). Impedance spectroscopy,

· :'==:d
0.2 (_ )0.6 0.6 I
John Wiley & Sons, New York.
Toyoda, K., H. Kojima, S. Miyarnoto and R
Takeuchi (1997). Measurement and analysis of
moisture changes in agricultural products using
FIT Noise Impedance Spectroscopy, Drying
0.2 0.4 ~ (_ )0.6 0.6 1 Technology, 15(6-8), pp.2025-2035,

~:I :.:=a
-76r---~-~--~-~--
Toyoda, K. (1994). Impedance spectroscopic
analysis in agricultural products
In:Developments in Food Engineering (yano,
Matsuno and Nakarnura), PART 1, pp.143-145,
-6°0 Blackie A & P, London,
0.2 0.4 ~ (_ )0.6 0.6

Zhang, M.I.N., D.G. Stout and 1.H.M. Willson(

1990). Electrical impedance analysis in plant
tissues: symplasmic resistance and membrane
Fig. 7 Relationship between the model parameters
capacitance in Hayden Model, Journal of
and frozen ratio in blast freezing
Expertmental BotanY,41(224).pp.371-380,
Zhang, M.I.N., and 1.H.M. Willson (199Ia).
4. CONCLUSIONS Electrical impedance analysis in plant tissues,
Can. 1. Plant Sci. 72,545-553,
Through the impedance measurements and analysis
Zhang, M.I.N. and J.H.M, Willson (199Ib).
of a freezing process of apple fruits, the following
Electrical impedance analysis in plant tissues:
findings were obtained:
a double shell model, Journal of
• The beginning and the end of the freezing
Experimental Botany, 42(244),pp. 1465-1475
could be detected by the extracellular
resistance. Zhang, M.I,N., D.G. Stout and J.H.M. Willson
• Three model parameters Ra, Rs and Cm (1992). Plant tissue impedance and cold
correlated well with the frozen ratio defined in acclimation: a re'analysis, Journal of
this study and these values were useful to Experimental Botany, 43 {247), pp. 263-266
estimate the frozen ratio.
Zhang, M.I.N., 1.H.M. Willson, M.A. Cox, and S.A.
• A time lag of about 120 seconds between the
Hall (1993). Measurement of heat injury in
rising of the cwves of intracellular resistance
plant tissue by using electrical impedance
and extracellular resistance. This suggested
analysis, Can. 1. Bot., 71 ,pp.1605-1611.
that the sequence of ice formation within the
cell tissues, namely extra- and intra-eellular
parts might be detected by the impedance
analysis.

, '[2]z::[ZJ//'//.
z::0//z::
~04
x,./
0.2
/",/ ~O.4
x
0.2
.
.~//
it
I

EO.4

0.2 /
",...'
/0/

o 0 0 •.
o . 0.5, \ 1 " . 0.5" 1 0 , 0.5" t

Fig. 8. Relationship between normalized parameters

and frozen ratio

94