ANALYTICAL SCIENCES JUNE 1989, VOL.

5 245

Reviews

Polymerization of Silicic Acid

A Review

Toshikazu TARUTANI

Department of Chemistry, Faculty of Science, Kyushu University, Hakozaki, Fukuoka 812, Japan

Useful information on the polymerization of silicicacid is obtained from a combined use of (1) spectrophotometry
based on the formation of yellow molybdosilicic acid, (2) trimethylsilylationof silicicacid and determination of
trimethylsilylderivatives by gas-liquid chromatography and (3) gel permeation chromatography of silicicacid. The
polymerization of silicicacid is divided into three processes: (1) the formation of polysilicic acid by the reaction
between molybdate reactive silicic acids (MRSacid), (2) the particle growth of polysilicicacid by the reaction
between MRS acid and polysilicic acid, and (3) the particle growth of polysilicic acid by the reaction between
polysilicicacids. The pH value giving maximum rate of polymerization of silicicacid is different among these three
processes. Fluoridepromotes processes (1) and (2); the effect of fluoride on the process (3) is relativelysmall.

Keywords Polymerization, silicic acid, spectrophotometry, trimethylsilylation-gas-liguid chromatography, gel per-

meation chromatography, catalytic effect, fluoride

1 Introduction 245 3.3 Gel permeation chromatography

2 Solubility of Silica 245 4 p H Value Giving Maximum Rate
3 Polymerization of Silicic Acid 246 of Polymerization of Silicic Acid 249
3.1 Spectrophotometry 5 Catalytic Effect of Fluoride on the
3.2 Trimethylsilylation-gas-liquid Polymerization of Silicic Acid 251
chromatography

production of silica sols or gels. More dilute systems

1 Introduction have been employed for studying the conditions
governing the polymerization of silicic acid. The
Monosilicic acid in aqueous solution reacts with factors that control the rates of polymerization of silicic
molybdate reagent under acidic conditions to form acid are the pH, temperature, degree of supersaturation
yellow molybdosilicic acid. This is a basic reaction for and presence of other substances. This review will
the spectrophotometric determination of monosilicic present some information concerning the polymeriza-
acid. Studies of silicic acid concentrations in natural tion of silicic acid and related problems.
water have revealed that concentrations are well below
the solubility of amorphous silica. Since silicic acid in
most natural water is of the monomeric form, its 2 Solubility of Silica
concentration can be determined by the spectrophoto-
metric method. However, an analytically important The solubility of silica in water is a basic factor in the
fact is that at the appropriate pH monosilicic acid in behavior of silicic acid. When solid phases of silica are
supersaturated solutions, such as geothermal or hot in contact with water, small amounts of silica dissolve
spring water, will in time polymerize, thus the exact and enter the aqueous phase. The dissolution and
value of the total silicic acid concentration in aged deposition of silica in water involve both hydration and
solutions can not be obtained by the spectrophoto- dehydration reactions:
metric method.
The polymerization of silicic acid in aqueous solution (SiO2)n+ 2H20 ::::: (SiO2)n-1+ S1(OH)4
has been studied for a long time. Much of the work
focused on reactions in which the initial monosilicic Here, (Si02) means solid silica or polysilicic acid in
acid concentrations were relatively high and on the water. At equilibrium, the concentration of monosilicic
246 ANALYTICAL SCIENCES JUNE 1989, VOL. 5

"Temperature/ -U
Fig. 2 Solubility of amorphous silica vs. p H (25° C).
Fig. 1 Solubility of amorphous silica.

acid in the aqueous phase, determined by the molyb-

date method, is the solubility of silica.
The solubility of silica is dependent on many factors
such as the state of the silica, whether it is in crystalline
or amorphous form, pH of the solution, temperature
and presence of other ions. The solubility of the
crystalline forms of silica is low and need not be
considered in analytical chemistry since the behavior of
silicic acid in aqueous solution is dependent on the
solubility of amorphous silica at temperatures lower
than 100°C. The solubility of amorphous silica in
water up to 100° C was reported by Kitahara (Fig. 1).1
The solubility of amorphous silica is determined in two
ways. When amorphous silica powder is suspended in
water the concentration of monosilicic acid increases to
Fig. 3 Variation of the concentration of MRS acid with pH.
an equilibrium solubility value. As polymerization
Initial monosilicic acid concentration: 500 ppm (SiO2).
proceeds in a supersaturated system, the monosilicic
acid concentrations decrease, approaching the solubility
of amorphous silica. However, different values for the
solubility have been reported in the literature.2 The
variations in the solubility values are due to differences obtained by these methods is briefly reviewed.
in the solubility characteristics of various form of silica.
The solubility increases with decreasing particle size of 3.1 Spectrophotometry
amorphous silica.3 The kinetics of polymerization of monosilicic acid
The solubility of amorphous silica in aqueous solu- have usually been studied by spectrophotometry, based
tions at different pH values at 25°C is shown in Fig. 2.4 on the decrease in the concentration of monosilicic acid
The solubility of amorphous silica barely changes in the with time. This could be done because monosilicic acid
pH range 1-9, but increases rapidly above pH 9, reacts with molybdate reagent to give a yellow color,
because of the formation of silicate ions. whereas polysilicic acid does not react with this reagent.
The solubility decreases with increasing salt concen- During the polymerization of monosilicic aicd, low-
trations molecular-weight silicic acids, such as dimer and trimer,
form. They then depolymerize and react with molybdate
reagent. Hereafter, silicic acids which can be measured
3 Polymerization of Silicic Acid by molybdate method are called molybdate reactive
silicic acid (henceforth abbreviated MRS acid), and
Investigations on the polymerization of silicic acid silicic acids which do not react with molybdate reagent
have been made by various methods. Among the are called polysilicic acid.
methods, spectrophotometry, trimethylsilylation-gas- The polymerization of silicic aicd is strongly pH
liquid chromatography and gel permeation chromatog- dependent. In Fig. 3, the concentration of MRS acid
raphy are useful for studying the polymerization of after 24 and 75 h is plotted against pH.6 In a strongly
silicic acid. Information concerning the polymerization alkaline solution monosilicic aicd does not polymerize.
ANALYTICAL SCIENCES JUNE 1989, VOL. 5 247

TMS derivatives have been prepared by treating a

silicate solution with a mixture of water, hydrochloric
acid, isopropyl12 or propyl13 alcohol and hexamethyl-
disiloxane. The TMS derivatives of low-molecular-
weight silicic acids can be identified'4 and quantified'5
by gas-liquid chromatography (GLC). The distribution
of derivative species reflects that which exists in the
parent silicate solution.
Figures 5, 6 and 7 show the variation of concentra-
Fig. 4 Variation of the concentration of MRS acid with time tions of silicic acid species with time.16 Each silicic acid
at pH 8.5. Initial monosilicic acid concentration: 600 ppm species is expressed in its anionic form. The Si(C) value
(SiO2). is the concentration of silicic acid determined by
spectrophotometry, and the Si(T) value is the sum of
the concentrations of each silicic acid species whose
TMS derivatives can be determined by GLC. The
The MRS acid concentration decreases rapidly in difference between the initial concentration of mono-
slightly alkaline solutions, but does not decrease within silicic acid and the Si(T) value indicates the amount of
24 h within the pH range 1- 3. Below this range the GLC-undeterminable silicic acid species.
rate of polymerization increases with greater acidity.' At pH 1.8 (Fig. 5), Si2076- and 5i30108 formed.
Figure 4 shows the effect of temperature on the During the first hour the amounts of 5i2076 and
polymerization of silicic acid at pH 8.5. At 20° C the Si30,08_increased with rapid decrease in the amount of
MRS acid concentration abruptly decreased in 1 h. SiO44~,but after this period did not change. This shows
After 2 h its concentration only decreased very slowly. that after 1 h an equilibrium is established among
On the other hand, at 90° C the MRS acid concentra- these silicic acid species. The Si(T) and Si(C) values did
tion was identical with the initial monosilicic acid not change with time and were equal to the initial
concentration until 4 h and then suddenly began to concentration of monosilicic acid. This indicates that
decrease. There exists an "induction period" during no GLC-undeterminable silicic acid species other than
which there is no change in the concentration of MRS MRS acids formed.
acid. Induction periods for the polymerization of MRS In the 2500-ppm solution at pH 1.8 (Fig. 6), silicic
acid from geothermal waters have been reported.8 The acid species other than Si2076- and 5i30,08, such as
induction period is greater at higher temperatures and S13096-, 5i40,310-, Si501S'o- and Si601710-formed pro-
decreases with increasing degree of supersaturation of gressively. The values for Si3096- and Si401310-were
monosilicic acid9 and salt concentration.10
The rate of decrease in the MRS acid concentration
increases with increasing salt cocnentration."
One disadvantage of the spectrophotometric method
is that it yields no information on the change in particle
size of polysilicic acid with time.

3.2 Trimethylsilylation-gas-liquid chromatography

Lentz12 described a chemical technique for studying
mineral silicate structures based on the trimethylsilyla-
tion of silicic acids liberated from minerals by acid
leaching. Trimethylsilylation studies provide valuable
information about silicate solutions.
Low-molecular-weight silicic acid species are convert-
ed into relatively unreactive trimethylsilyl (TMS) de-
rivatives, e.g. as in Eqs. (1) and (2), where X=Si(CH3)3.
OH OX
I trimethylsilylation I Fig. 5 Changes in the concentration of silicic acid species in
HO-Si-OH XO-Si-OX (1) the solution of pH 1.8 with time. Initial monosilicic acid
OH OX concentration: 600 ppm (SiO2).
248 ANALYTICAL SCIENCES JUNE 1989, VOL. 5

with time can be studied by gel permeation chromatog-

raphy.

3.3 Gel permeation chromatography

One method for separating solute molecules accord-
ing to their size is gel permeation chromatography
(GPC), the basic principles of which have been
explained in terms of a sieving effect or steric exclusion.
The separation of the components of a sample in a
solution is usually carried out on a column packed with
a gel or other porous material. The elution volumes of
molecules are correlated with their molecular size,
larger molecules being eluted faster than smaller
molecules.
Polysilicic acid can be separated from MRS acid by
GPC.17 Solutions of silicic acid for GPC are usually
adjusted to pH 2 at which polymerization does not
occur for several hours or days.6
The distribution coefficient, Kay, for a given gel in
GPC is defined's as

Kav - (Ve V0)/(V-t V0)

where Vo is the void volume of the gel bed, Vt the total

bed volume and Ve the elution volume of the solute.
Va, Vt and Ve are easily measured. Since Kay is
correlated to Ve, it is a measure of the molecular size,
decreasing with increasing particle size. All polymers
that have larger sizes than the possible separation range
of the gel are eluted at V0.
Figure 8 shows typical elution curves for silicic acid,
obtained at pH 9, 8 and 6 on a Sephadex G-100.6 The
peaks on the right are due to MRS acid and those on
the left to polysilicic acid. The concentration of MRS
acid at a given time is shown in parentheses.
At pH 9.5 the elution curves for polysilicic acids
indicate a symmetrical distribution of particle sizes; the
curve for polysilicic acids obtained after 250 h was
almost the same as that obtained after 100 h. This
Fig. 7 Changes in the concentration of silicic acid species in means that when the concentration of MRS acid is
the solution of pH 8.5 with time. Initial monosilicic acid similar to the solubility of amorphous silica, the growth
concentration: 600 ppm (SiO2). rate of the particles of polysilicic acids becomes very
slow. Results indicate that the growth of polysilicic
acids is due mainly to polymerization between MRS
acid and polysilicic acid, and that polymerization
not plotted in the figure because they were insignificant. between polysilicic acids hardly proceeds. Similar
Although the Si(T) value gradually decreased with time, results were obtained at pH 9 and 2.
the Si(C) value was equal to the initial concentration of At pH 6 the elution curve for polysilicic acids
monosilicic acid and did not change with time, obtained after 24 h indicates a broad and random
indicating that all of the GLC-determinable and the distribution of particle sizes. After 75 h the peak of
GLC-undeterminable silicic acid species formed were polysilicic acids appeared at V0. This indicates that
determined by spectrophotometry. The results shown particles of polysilicic acids grow rapidly. The broad
in Figs. 5 and 6 indicate the formation of origomeric and random distribution of particle sizes arises from a
silicic acid species during the induction period. random condensation of polysilicic acid species. Simi-
At pH 8.5 (Fig. 7), the amounts of Si044-, Si2076- lar results were obtained at pH 7 and 4.
and Si301oa-, and Si(T) and Si(C) values abruptly At pH 8 the elution curve for polysilicic acids
decreased in 1 h. This indicates a rapid formation of obtained after 2 h was symmetrical. The MRS acid
polysilicic acids. concentration after 75 h was not much different from
The variation of the particle sizes of polysilicic acids that after 24 h and was near the solubility of amor-
ANALYTICAL SCIENCES JUNE 1989, VOL. 5 249

Fig. 9 Plot of Kay values of polysilicic acid against temper-

ature.

(1) Formation of polysilicic acid by a reaction between

MRS acids (M-M reaction).
(2) Particle growth of polysilicic acid by a reaction
between MRS acid and polysilicic acid (M-P
Fig. 8 Variation with time of the elution curves for silicic reaction).
acid in solutions of pH 9.5, 8 and 6. Initial monosilicic
acid concentration: 500 ppm (Si02). Gel: Sephadex G-100. (3) Particle growth of polysilicic acid by a reaction
between polysilicic acids (P-P reaction).
Column: 45X1.0 cm i.d. (1), pH 9.5: ---, 6 h (M: 219 ppm);
-
, 100 h (M: 192 ppm); ...•, 250 h (M: 174 ppm). (2),
If it can be assumed that in the early stage of MRS
pH 8: ---, 2 h (M: 230 ppm); -, 24 h (M: 147 ppm); ...•,
75 h (M: 136 ppm). (3), pH 6: ---, 24 h (M: 298 ppm); -, acid polymerization the decrease in the concentration of
75 h (M: 186 ppm). MRS acid in solutions at various pH values is due
mainly to an M-M reaction, it may be possible to obtain
a relationship between the rate of the M-M reaction
and the pH. The pH of the maximum of the decrease
phous silica, though particle growth still continued and in the concentration of MRS acid after short period of
the elution curves became asymmetrical. These results time is the pH at which the rate of the M-M reaction is
indicate that in the early stage of polymerization the maximum. The kinetics of polysilicic acid formation
growth of particles is due mainly to polymerization obtained by monitoring the concentration of MRS acid
between MRS acid and polysilicic acid, though the as functions of the time and the pH are shown in
polymerization between polysilicic acids is subsequently Fig. 10. It is considered that the pH of the maximum
predominant. of the decrease in the concentration of MRS acid after
The rate of particle growth of polysilicic acids a few minutes may be close to the pH at which the rate
increases with increasing sodium chloride concentra- of the M-M reaction is maximum. The pH giving the
tion.11 Crerar et a1.19reported that divalent metal salts maximum rate of decrease in the concentration of MRS
(MgCl2, CaCl2, SrCl2) promote the particle growth of acid shifts slightly to a lower pH with the lapse of time.
polysilicic acids more than does NaCI. This may be due to the influence on polysilicic acid
The rate of particle growth of polysilicic acid by the formation by the M-P reaction. The results indicate
polymerization between polysilicic acids is affected that at 20° C the M-M reaction is most rapid at around
with temperature. Figure 9 shows the relationship p H 9.3.
between Kay values of polysilicic acids, calculated from When MRS acid solution in which the M-M reaction
the elution volumes, in solutions of pH 7 after 1 h and does not occur within a certain period of time is mixed
temperature.20 The Kay value of polysilicic acid in with a polysilicic acid solution and the concentration of
initial solutions was 0.52. The concentration of MRS MRS acid decreases with time from the moment of
acid in each solution was nearly equal to the solubility mixing of the solutions, the decrease in the concentra-
of amorphous silica at given temperature. The Kay tion of MRS acid is due mainly to the M-P reaction.
value of polysilicic acid decreases with temperature. The pH of the maximum of the decrease in the concen-
This result shows that the rate of particle growth of tration of MRS acid is the pH at which the rate of the
polysilicic acid increases with increasing temperature. M-P reaction is maximum. The procedure for the
determination of the M-P reaction has been carried out
4 pH Value Giving Maximum Rate of Poly- under these conditions. Results are shown in Fig. 11.
merization of Silicic Acid The decrease in the concentration of MRS acid was
maximum at around pH 8.6 at 20° C.
The polymerization of silicic acid is divided into three When the concentration of MRS acid is close to the
processes. solubility of amorphous silica at a given pH, the M-M
250 ANALYTICAL SCIENCES JUNE 1989, VOL. 5

Fig. 10 Variation of the concentration of MRS acid as func-

tions of pH and time at 20° C. The horizontal line shows
initial monosilicic acid concentration. Time after adjusting
pH: 5 min (•); 10min (cP); 15 min (0). Fig. 12 Variation of the elution curves for polysilicic acid
after 48 h in solutions of various pH at 20° C. Gel, Sephadex
G-100; column, 45X1.5 cm id.; initial polysilicic acid con-
centration, 350 ppm (Si02). (1), pH 2 (-); pH 7 (----);
pH g (....); pH 9.5 (---); pH 10 (---). (2), pH 1 (---);
pH 2, 3 (-); pH 4 (---); pH 5 (....); pH 6 (----).

Fig. 11 Change in the concentration of MRS acid with pH

after 30 min at 20° C when polysilicic acid was added to a
monosilicic acid solution. Initial concentrations: monosilicic
acid, 314 ppm (Si02); polysilicic acid, 187 ppm (Si02).

Fig. 13 Plot of Kay values of polysilicic acid against pH.

and M-P reactions virtually do not occur. Since the

P-P reaction proceeds regardless of the presence of
MRS acid, the effect of pH on the P-P reaction can be acid in initial solutions of various pH. At pH>4 the
measured by GPC from the change in the elution curves elution curves obtained after 48 h were shifted to the
of silicic acid in solutions of various pH. The pH of the left compared with that at pH 2. This means that the
maximum of the decrease in Kayof polysilicic acid is the particle sizes of polysilicic acids were larger than under
pH at which the P-P reaction is maximum. An aliquot the initial solutions. Figure 13 shows the relationship
of a solution containing polysilicic acid, of which the between the Kay values of the polysilicic acids and the
particle size distribution was known, was added to pH. The Kay value is minimum at around pH 6.8 at
monosilicic acid solution. The monosilicic acid con- 20° C. This means that the P-P reaction is most rapid
centration in each sample solution was adjusted to be at this pH.
near the solubility of amorphous silica at the given The pH values at which the M-M, M-P and P-P
pH.21 Figure 12 shows typical elution curves for silicic reactions at different temperatures are most rapid are
acid obtained after 48 h in solutions of various pH. listed in Table 1.20 Polymerization due to three
The elution curve at pH 2 did not change within 48 h, reactions mainly proceeds by a reaction between
and represents the particle size distribution of silicic ionized and un-ionized silanol groups. The pH values
ANALYTICAL SCIENCES JUNE 1989, VOL. 5 251

Table 1 The p H values for which M-M, M-P and P-P

reactions are most rapid

Fig. 15 Elution curves for silicic acid and fluoride in solution

of pH 1 after 4 d (R=0.5): (0) Si; (S) F.

Fig. 16 Variation with time of the elution curves for silicic

acid in a solution of pH 1(R=0.1). Initial monosilicic acid
concentration, 500 ppm (Si02); gel, Sephadex G-100; col-
umn, 45X1.0 cm i.d. Time: 3 d (M: 343 ppm); --- 5 d (M:
262 ppm); --- 10 d (M: 198 ppm); - 17 d (M: 177 ppm).
Fig. 14 Plot of Kay values of polysilicic acid after 5 h against
the R value. Initial monosilicic acid concentration: 500 ppm
(SiO2). pH: 0.55 (0); 1 (a); 2.5 (0).
was most rapid at about pH 1.
Elution curves for silicic acid and fluoride in solution
(R=0.5) at pH 1 after 4 d are shown in Fig. 15. The
giving the maximum rates of the M-M, M-P and P-P elution volume of fluoride is equal to that of MRS acid,
reactions shifted to lower pH values with increasing and polysilicic acid is not accompanied by fluoride.
temperature. This may have been due to a change in The variation with time of the elution curves for
the ionization constants of silicic acid species. silicic acid in solution (R=1) at pH 1 is shown in Fig.
16. The concentration of MRS acid is shown in paren-
theses. The elution curve for polysilicic acids after 5 d
5 Catalytic Effect of Fluoride on the Polymeriza- indicates a nearly symmetrical distribution of particle
tion of Silicic Acid sizes. The elution curve obtained after 17 d was almost
the same as that obtained after 10 d. These results
The gelling of silicic acid sols at low pH is markedly indicate that when the MRS acid concentration is close
catalyzed by traces of fluoride.22 The rate of decrease to the solubility of amorphous silica the growth rate of
with time of the concentration of MRS acid in an acidic polysilicic acid particles is very slow in spite of the
solution in the presence of fluoride is faster than that in presence of fluoride. It is concluded that the growth of
the absence of fluoride.23 The rate of decrease in the polysilicic acid particles is due mainly to the M-P
concentration of MRS acid increases with decreasing reaction, and that the catalytic effect of fluoride on the
pH. The rate of the M-M reaction in solutions in the P-P reaction is relatively small.
pH range 2.5 -0 is most rapid at a F/ Si atomic ratio, R, Fluoride prevents the color development of molyb-
of about 0.5, regardless of the difference in acidity.24 dosilicic acid in spectrophotometry. Aluminum, borate
In Fig. 14 the Kay values of polysilicic acids in and beryllium ions which combine with fluoride to
solutions of various pH after 5 d are plotted against the form fluoride complexes have a masking effect for
R value. In silicic acid solutions containing fluoride (up fluoride and aluminum ion most effectively masks
to R=2.0) the rate of growth of polysilicic acid particles fluoride.25 Similary, the influence of fluoride on the
increased with increasing fluoride concentration and polymerization of silicic acid can be overcome by the
252 ANALYTICAL SCIENCES JUNE 1989, VOL. 5

addition of aluminum ion to give the desired atomic Kyushu Univ., Ser C, 13, 303 (1982).
ratio of aluminum and fluoride (Al/ F=10).26 14. G. Garzo and D. Hoebbel, J. Chromatogr., 119, 173
(1976).
15. T. Shimono, T. Isobe and T. Tarutani, J. Chromatogr.,
References 205, 49 (1981).
16. T. Shimono, T. Isobe and T. Tarutani, J. Chromatogr.,
258, 73 (1983).
1. S. Kitahara, Rev. Phys. Chem. Jpn., 30, 131 (1960).
17. T. Tarutani, J. Chromatogr., 50, 523 (1970).
2. R. K. Iler, "The Chemistry of Silica'; p. 40, John Wiley &
18. T. C. Laurent and J. Killander, J. Chromatogr., 14, 317
Sons, New York, 1979.
3. G. B. Alexander, J. Phys. Chem., 61,1563 (1957). (1964).
19. D. A. Crerar, E. V. Axtmann and R. C. Axtmann,
4. G. B. Alexander, W. M. Heston and R. K. Iler, J. Phys.
Geochim. Cosmochim. Acta, 45,1259 (1981).
Chem., 58, 453 (1954).
20. K. Shimada and T. Tarutani, Mem. Fac. Sci., Kyushu
5. W. L. Marshall and J. M. Warakomski, Geochim.
Univ., Ser C, 13, 311 (1982).
Cosmochim. Acta, 44, 915 (1980).
21. K. Shimada and T. Tarutani, Bull. Chem. Soc. Jpn., 53,
6. K. Shimada and T. Tarutani, J. Chromatogr., 168, 401
3488 (1980).
(1979). 22. R. K. Iler, J. Phys. Chem., 56, 680 (1952).
7. H. Iwasaki, K. Shimada and T. Tarutani, J. Chromatogr.,
23. T. Tarutani, Nippon Kagaku Zasshi, 77, 1811 (1956).
198, 429 (1980).
24. K. Shimada and T. Tarutani, J. Chromatogr., 249, 111
8. D. E. White, W. W. Brannock and K. J. Murata,
Geochim. Cosmochim. Acta,10, 27 (1956). (1982).
25. T. Tarutani, Nippon Kagaku Zasshi, 77,1292 (1956).
9. H. Bauman, Kolloid Z., 162, 28 (1959).
26. T. Yokoyama, H. Kitsuki and T. Tarutani, J. Jpn.
10. A. C. Makrides, M. Turner and J. Slaughter, J. Colloid
Geothermal Energy Assoc., 20, 316 (1983).
Interface Sci., 73, 345 (1980).
11. Y. Takahashi, T. Yokoyama and T. Tarutani, J.
(Received February 15, 1989)
Geothermal Res. Soc. Jpn., 10, 225 (1988).
12. C. W. Lentz., Inorg. Chem., 3, 574 (1964). (Accepted March 16, 1989)
13. T. Shimono, T. Isobe and T. Tarutani, Mem. Fac. Sci.,