Studies on the Relationship Between
Water-satured State and Crystallinity
by the Diffraction Method for
Moistened Potato Starch
Crystallinity was measured by X-ray diffraction method and water-
saturated state was determined by means of the moisture contents
calculated from the crystallinity for moistened cellulose and potato
starch. In the X-ray diffractograms the upper areas which were
separated by the straight line joined the points of intensity at 37" and
4" (8") of diffraction angle was the relative reflection area in the
crystalline and amorphous portions, and the lower area was the
background of the non-relative reflection area. The upper area
separated by the smooth curve joined the minimum intensity at 4" to
37" of diffraction angle corresponded to the crystalline portion and
the lower area to the amorphous one. The ratio of the crystalline
portion to sum of the crystalline and amorphous portions was
considered to be crystallinity. Crystallinity of the air-dried and the
moisture-saturated (28%) samples of cellulose were 0.68 and 0.71,
resp. The crystallinity of the air-dried and water-saturated (49%)
samples of potato starch were 0.24 and 0.32, resp.
1 Introduction
The authors reported on crystallinity by the X-ray diffraction
method [l], moisture regain [2], specific volume [3] and
deuteration [4], and also did sorption phenomena [5] and acid
resistance [6] for starch granules. Since crystallinity of starch
granules increased with an increased sorption water, it seemed
to become maximum in water-saturated state. The water-
saturated state of starch granules was given by the vapor [7,8]
and centrifuging methods [9], but determination of the
saturated state has not been made clear yet. Moreover, a
general determination method of crystallinity of starch was
not elucidated because the 100% and various ratio crystalline
starches were unknown.
Consequently, crystallinity of starch granules was expressed
by the relative crystallinity and the crystal index [l, 103. The
authors discussed the measurement of crystallinity of starch
granules by separating the crystalline and amorphous
portions in X-ray diffractogram and moreover, did the
determination of water-saturated state on the basis of
crystallinity.
2 Materials and Methods
2.1 Air-dried materials
The starch sample used was commercial potato starch refined
with cold alkaline solutions by the same manner as described
in the previous report [l] and the refined materials dried in
starch/starke 35 (1983) Nr. 12, S. 407-410 0 Verlag Chemie GmbH, D-6940 Weinheim 1983
0038-9056/83/1212-0407$02.50/0
By S. Nara and T. Komiya, Tsu (Japan)
~ ~~
Untersuchungen uber die Beziehung zwischen Wassersattigung
und Kristallinitat mittels der Beugungs-Methode fur feuchte
Kartoffelstarke. Die Kristallinitat von feuchter Cellulose und
Kartoffelstarke wurde mittels der Rontgenbeugungs-Methode, und
der Wassersattigungs-Zustand wurde mit Hilfe der aus den Feuchtig-
keitsgehalten errechneten Kristallinitat bestimmt. In den Rontgen-
beugungs-Diagrammen waren die oberen Bereiche, die durch die
gerade Linie zwischen den Intensitatspunkten 3 7 und 4" (8")
Beugungswinkel getrennt waren, der relative Reflexionsbereich der
kristallinen und amorphen Anteiie, und der untere Bereich warder
Hintergrund des nichtrelativen Reflexionsbereiches. Der obere
Bereich, der durch die flache Kurve die minimale Intensitat bei 4" bis
37" Beugungswinkel abgetrennt war, entsprach dem kristallinen und
der untere Bereich dem amorphen Anteil. Das Verhaltnis des
kristallinen Anteils zu der Summe aus kristallinem und amorphen
Anteil wurde als Kristallinitat angesehen. Die Kristallinitat der
luftgetrockneten und der feuchtigkeitsgesattigten (28YO)Cellulose-
proben betrug0,68 bzw. 0,71. Die Kristallinitlt der luftgetrockneten
und wassergesattigten (49%) Kartoffelstarkeproben betrug 0,24
bzw. 0,32.
atmosphere were passed through a 200 mesh screen. The
cellulose sample was the powder one for chromatography
manufactured by Whatman Co. and was sieved through a 100
mesh screen.
2.2 Moistened materials
Preparation of moistened materials was done by the vapor
and the centrifuging methods [8,9]. The vapor method was as
follows: About 10 g of air-dried samples in a dish of 8.5 cm
diameter were placed in the desiccator where at the bottom
appropriate amounts of water were placed under low pressure
at room temperature. The samples were stirred some times
with spatula and sorbed water vapor for 10 to 30d. The
centrifuging method was as follows :About 1 g of the air-dried
sample was put the centrifugal filter tube fitted with a filter
paper (Toyo Filter No. 2 ) at the bottom and sorbed water
added in the tube at room temperature. After 24 h, the water-
sorbed sample was hydroextracted by centrifuging for 50 min
using the p F measured centrifuge (No. 55 - 1)manufactured
by Sakuma Manufacture Co. Japan. The centrifugal effect
was expressed by the value of pF, that is, the logarithm of
centrifugal effect ( g ) . For example, pF 4 is log lo4 (8).
The p F value was calculated from the equation (1) and the
relative humidity at p F value was calculated from the
equation (2) [9, 113:
407
 +
pF = 4.036 log (2 -log RH) + log (273 + t ) (a,
where r =radius, 6.307 cm, g = gravity, n = revolutions per
min, RH = relative humidity (%), t = temperature ("C).
2.3 Moisture content
The moisture content of starch was measured by vacuum
drying at 120°C for 4 h and that of cellulose was done by
drying at 105°C. The moisture content was expressed as the
value per dried materials (dry basis).
2.4 X-ray diffraction method
The X-ray diffraction method was almost the same as
described in the previous reports [l, 61. Copper K radiation
passed through a Ni filter was generated by a Rigaku Denki
X-Ray Diffractometer D-3F type. The cellulose and starch
powders were held in an aluminum cell (thickness 0.15 cm)
and exposed to the X-ray beam (30 KV, 15 mA) with the
symmetrical reflection technique. Both width of the divergen-
ce and scatter slits were 1" and that of the receiving slit was
0.4 mm. The scanning regions of the diffraction angle were 4"
to 37" of 2 8. The chart speed was 2 cm/min and 1 cm of chart
width corresponded to 1"of 2 8. The height of full scale on the
chart was 24cm and the maximum intensity was 1600
counts/min for starch and 4000 counts/min for cellulose.
the equation (3) was 0.025 for cellulose and 0.004 for potato
starch at 37" of 2 8. The value of 0.004 was considered to be
negligible. The value of 0.025 was slightly large to be
neglected, but the value was considered to be negligible
because it decreased with a decreased 8. The normalization of
intensity was not done because of IN= I, for cellulose and
potato starch.
+
The ratio of the each value of a, a,and ac/u,(u, = a, a,)of
the moistened sample to that of the air-dried one was
calculated to give the relative crystallinity (in this case, a
decrease of a, meant an increase of crystallinity).
3 Results
The Figures 1 and 2 show the diffractograms ofthe moistened
cellulose (by centrifuging at pF 3.93) and potato starch (by
vapor processing for 30 d where the separation methods of
their crystallinity (a,)
and amorphous (a,) portions are given.
Table 1 shows the moisture contents, and the relative
humidity in a. a, and a,/at for the moistened cellulose and
potato starch which were prepared by the vapor processing
for 10 and 30 d and by centrifuging at pF 3.93 to 4.20.

2.5 Relative crystallinity

The measurement of the relative crystallinity to which the
Hermans method [12 - 171was applied was done according to
the same manner as reported previously [1,6]. The separation
methods of the crystalline and amorphous portions in the
X-ray diffractograms of cellulose and potato starch were
shown in Figures 1 and 2. The upper area (a,)which was
L" 10' 15' 20" 25" 30" 35"37"
separated with the smooth curve connecting each point of the Diffraction angle(2 8 )
minimum intensity corresponded to the crystalline portion
and the lower area was background containing the amor- Figure 1. X-Ray diffractogram of cellulose hydroextracted by
phous portion (a,). a, was assumed to be the upper area centrifuging at pF 3.93. u, and u, indicate the crystalline and
separated with the straight line joining the two points of amorphous portions in the X-ray diffractogram, resp.
intensity at 37" and 4" (8") in the background. By assuming
that the all integral intensity from 4" to 37" was constant, the
areas of a,and a,were corrected to be the value of250cmz as
described in the previous report [6]. Correction of air
scattering and utilization ratio of X-ray first beam was not
done.
The normalized intensity (IN)of reflection intensity (I,) was
expressed as follows :

1-exp (-2 Dcosec 8)

1-'
where PIP, = corrected value of first beam intensity, D =
X-ray optical density.
The initial and terminal intensity ofthe first beam were almost
constant by checking with astandard ofsilicon powder and as
a result, it was assumed to be P, = P.
The values of the linear absorption coefficient (p) [12] for
cellulose and potato starch were 12.03and 11.75, resp., which
were caluculated with the respective density (y)of 1.551 and
1.517 [3,5]. The apparent density (4.1') of cellulose and potato
starch wasOSOand0.75,resp. and thickness was0.15 cm. The
optical density of cellulose and potato starch was 0.58 and
0.87, resp. which was calculated from the equation, D = p't
Figure 2. X-Ray diffractogram of potato starch moistened by the
vapor processing for 30 d.
In the case of cellulose, when each relative crystallinity of the
air-dried material in a, aaand a,fa, was assumed to be 1, the
relative crystallinity of the moistened one having the values of
0.90 to 0.95 was slightly smaller than that of the air-dried one
in a,.But the relative values having 1.01 to 1,06 and 0.78 to
0.87 in a,/a, and a,,respectively were slightly larger than the
respective relativecrystallinity ofthe air-dried one in ac/u,and
a,.For the cellulose sample moistened in atmosphere for 15
after dehydration in a desiccator containing P,05 at room
temperature, the relative crystallinity in a, was 1.06 and

= (t. ') .
y t. The maximum value ofthe exponential part in
slightly smaller than that ofthe air-dried one in a,,but those in
u,and a,/a, were 1.09 and 1.01, resp. and were slightly larger
than those of the air-dried one in a,and a&,.

408 starch/starke 35 (1983) Nr. 12, S . 407-410

Table 1.
Moisture Contents and the Relative Crystallinity of the Moistened Cellulose and Potato Starch.Samples : Moistened Cellulose and Potato Starch.
a, and an.Indicate the Crystalline and Amorphous Portions in the X-Ray Diffractograms of the Samples, resp., at = a, + aa.

Moistened Vapor Method Centrifuging Method

Air dried after d 10 20 30 PF 3.93 4.00 4.08 4.20
Dehydration RH% 100 100 100 RH% 99.4 99.3 99.2 98.9

Cellulose
Moisture
Content (YO) 5.15 4.90 17.47 20.19 22.15 27.92 26.63 25.63 24.00
r c (cm2) 105.3 115.1 94.6 97.2 98.8 97.9 98.4 99.7 96.2
1 1.09 0.90 0.92 0.94 0.93 0.93 0.950 0.91
%a (cm2) 48.5 51.3 42.4 38.8 39.8 37.2 39.0 42.3 40.0
1 1.06 0.87 0.80 0.80 0.78 0.80 0.87 0.82
a,/at (ratio) 0.683 0.691 0.691 0.715 0.713 0.725 0.716 0.702 0.703
1 1.01 1.01 1.05 1.05 1.06 1.05 1.03 1.03

Potato Starch
Moisture
Content (%) 19.80 16.57 44.60 44.97 49.99 54.63 54.15 53.74 49.14
E c (cm2) 25.6 21.9 34.1 35.7 33.8 27.4 28.1 27.3 28.5
1 0.86 1.34 1.40 1.32 1.07 1.12 1.07 1.11
a, (cm2) 80.9 102.7 61.7 66.8 71.7 62.5 63.9 66.2 65.5
1 1.27 0.84 0.83 0.89 0.17 0.79 0.82 0.81
aJat (ratio) 0.240 0.176 0.335 0.348 0.320 0.304 0.310 0.290 0.303
1 0.73 1.40 1.45 1.34 1.27 1.29 1.22 1.26

From the results mentioned above, the crystallinity of
cellulose was not changed by the moisture drying and
absorbing as reported by Morsi [18]and recognized generally.
The crystallinity of cellulose was considered to be almost
constant, even if the moisture content of cellulose changed
from 17 to 28%.
In the case of potato starch, when each relative crystallinity of
the air-dried material in a, u, and uc/u,was assumed to be 1,
the relative values of the moistened materials in u, a, and
ctc/ct,were1.07to1.40,0.77to0.89and1.22to1.55,resp.and
were relatively larger than the corresponding relative
crystallinity of the air-dried one. The relative crystallinity of
the air-dried materials which sorbed moisture in atmosphere
for 15 d after having been dried in pz05 were 0.86,0.75and
1.27 in ct, uJu, and a,, resp. and all of them were relatively
smaller than the corresponding relative crystallinity of the so-
called air-dried one.
From these results, it was considered that the air-dried potato
starch which sorbed moisture only for 15 d in atmosphere
after having been dried in PzO5was not restored to the level of
the so-called air-dried one. Also,crystallinity of the moistened
potato starch seemed to have about similar values in the
moisture contents of 44 to 54%.
4 Discussion
4.1 Crystallinity
The studies on the crystallinity of starch by X-ray diffraction
method were reported only by Sterling [I8 -201 and Lelidvre
[21]and there were a few reports on the crystallinity. This is
because a general determination method of crystallinity of
starch was nlit elucidated, and 100% and various ratio
crystalline samples of starch were not obtained. The authors
studied the measurement of crystallinity of starch by means of
the separation of crystalline and amorphous portions from
background in the X-ray diffractogram and moreover, by
referring to the reports by Hermans [12,15 - 171.
By assuming that the areas ofthex-ray reflection intensity for
lOO%crystallineand 100%amorphous materials wereA,and
A,, respectively, Hermans proposed the relation between
these areas and crystallinity as follows: uc= x . A,, CL, =
(1 -x) .A,, a, and u, were the areas of crystalline and
amorphous portions in the X-ray diffractogram of a
crystalline polymer, resp.
In our experiments, it was assumed that the reflection
proportional constant for the crystalline portion in the
material was the same as for the amorphous portion because
the starch samples having 100% and various ratios of
crystallinity were unknown. When the areas of =,and 01, were
given as the following equations: u,= x . u,, u, = (1-x) . a,,
a, = a, + u,, the relation of A, = A. = a, was given from the
above assumption.
On the cellulose,the values of ct,/u, shown in Table 1 were 0.68
for the air-dried material and 0.69to 0.73 for the moistened
one. These values were almost the same as the crystallinity of
cellulose hitherto reported [21- 261.Therefore, the value of
u,/u, was considered to be crystallinity for cellulose.
On the potatostarch, thevaluesofu,/u,showninTable 1 were
0.24 for the air-dried material and 0.29 to 0.35 for the
moistened one. Each value was almost the same as the values
of 0.24and 0.28to 0.32in the references [l, 18,19,27].Thus,
the value of u,/u, for the potato starch was considered to be
crystallinity in the same manner as in the case of cellulose.
Moreover, the authors measured the value of u,/u, for the
Niigeli amylodextrin whose study was reported previously [6].
The Nageli amylodextrin was prepared as follows: l o g of
potato starch were hydrolyzed in 100ml of 16% sulfuric acid
at 30°Cfor 10 to 90 d. The acid treated material was washed
with distilled water and then hydroextracted by centrifuging
at pF 3.93 to 4.20.Table 2 shows the changes in the value of
u,/u, and the hydrolysis ratio for potato starch by hydrolysis
time. The values of u,/u, were about 0.34,0.43and 0.46at the
hydrolysis time of 0, 60 and 90 d, resp.
The hydrolysis ratios at a hydrolysis time of 60 and 90d were
1.25 and 1.33 times as many as that at hydrolysis time of 0 d,
resp. The increasing ratios of crystallinity at 60and 90d were
1.18 and 1.28 times as much as that of the original one,
respectively when calculated from the reciprocal of hydrolysis

starch/starke 35 (1983) Nr. 12, S . 407-410 409

Table 2.
Crystallinity of Niigeii Amylodextrin Prepared from Potato Starch by the Acid Hydrolysis Method. ac and ua Indicate the Crystalline and
Amorphous Portions in the X-Ray Diffractogram of Niigeli Amylodextrin, resp. Niigeli Amylodextrin was Prepared from Potato Starch by
Treatment with 16% Sulfuric Acid at 30°C.
Hydrolysis Time (d) 0 15 30 45 60 75 90
~

Hydrolysis ratio (%) 0 3.3 7.6 11.3 15.2 18.7 22.0

Hydrolysis residue (ratio) 1 0.967 0.924 0.887 0.848 0.813 0.780
l/Hydrolysis residue 1 1.04 1.08 1.13 1.18 1.23 1.28
a c (cm2) 37.5 43.9 45.3 47.9 48.3 48.8 50.5
zd. (crn’) 12.0 61.4 63.6 66.4 64.8 60.4 60.6
ciL ‘zt (ratio) 0.342 0.394 0.416 0.419 0.421 0.447 0.455
1 1.15 1.22 1.23 1.25 1.31 1.33
-

residue. The respective values of uc/ctt,1.25and 1.33 at 60 and Bibliography

90 days were very near t o the corresponding values of 1.18 and
1.28. From these results, the value of CI,/CI, for the Niigeli
amylodextrin was considered to be crystallinity in the same
manner as in the cases of cellulose and potato starch.
Consequently, crystallinity was 0.71 k 0.02 for the moistened
cellulose, 0.32 0.03 for the moistened potato starch and 0.46
for the Nageli amylodextrin.
4.2 Water saturated state
The water contents of the moistened cellulose samples
prepared by the vapor and the centrifuging methods [8, 91
varied from 17 t o 28% but crystallinity ofthe samples showed
the nearly constant value of 0.71 f0.02.
Also, the crystallinity of the moistened potato starch samples
gave the nearly constant value of 0.32 k 0.03 in spite of the
variation of the moisture content from 45 t o 55%. Conse-
quently, it was difficult t o determine the water-saturated state
of cellulose and potato starch from the crystallinity by the X-
ray diffraction method. The relationship between saturated
moisture content (R) and crystallinity (x) was shown by the
following equation (4) reported by Nara [2]:
+ 1 -x)
R = 0.0667 ( 0 . 1 8 ~ (4)
The saturated moisture content of cellulose was 0.279 when
calculated from the equation (4) with 0.71 of the average
crystallinity. The value of 0.279 agreed with the fiber
saturated point of 0.28 [28]. Since this value was given by
centrifuging at p F 3.93, it was considered t o be the moisture
content of cellulose in the water-saturated state. When the
moistened materials were dehydrated by centrifuging at more
than p F 3.93 and also when the air-dried ones sorbed moisture
by the vapor processing for 30d, these moisture contents
became smaller than that in the water-saturated state.
The saturated moisture content of potato starch was 0.49
when calculated from the equation (4) with 0.32 of the average
crystallinity. This value of0.49 was almost same as 0.509 and
0.493 of the saturated moisture content by means of the vapor
processing reported by Hellman [7] and Nara [9], resp.
Consequently, the water-saturated state of potato starch was
considered t o be obtained by the vapor Brocessing for 30 d and
by centrifuging at p F 4.20, since the saturated moisture
content of potato starch was given as 0.49.
[l] Naru, S., A . Mori and T. Komiya: Starch/Starke 30 (1978),111.
[2] Nara, S.:Starch/Starke 30 (1978),183.
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1948, translated in Japanese by Y . Noguchi and H . Fukuda,
Asakura (Tokyo) 1948,p. 222.
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1973,p. 64,74,pp. 150-155.
[I31 Hermans, P . H.,andA. Weidinger:J.Appl.Phys.19 (1948),491.
[I41 Hermans, P . H., and A. Weidinger: J. Polym. Sci. 4 (1949),135.
[IS]Hermans, P . H . , and A. Weidinger: Makromol. Chem. 44-46
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[21]Leliivre, J . : Starch/Starke 26 (1974),85.
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Elsvier, New York 1949,p. 316.
[23]Howsmon, J . H . : Textile Res. J. 19 (1949),152.
[24]Kast, W . : Z.Elektrochem. 57 (1953),525.
[25]Oda, R . , S. Makizima, M . Imoto, W . Sakai and Y . Iwakura:
“Modern Industrial Chemistry 22. Industrial Chemistry of
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[26]Sobue, H., and S . Fukuhara: J. Chem. SOC.Japan, Ind. Chem.
Sect. 60 (1957),32.
[27]Zobel, H . F., and R . Senti: “Program of 45th Annual Meeting
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Wood Technol. (2)”,Mc. Graw Hill, New York 1952,p. 26.

Address of authors: Prof. Dr. Shozo Nara and Dr. Takashi Komiya,
Mie University, Faculty of Agriculture, Tsu, Mie-ken, 514 Japan.

(Received : February 18, 1983)

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41 0 starch/stlfke 35 (1983)Nr. 12,S. 407-410