7-ol-----l
LMW
-Tr=
AMYLOIE \ I
I I
1 I I VI I
0 0.11 0.4 0.6 0.8 1.0 1.2
I
r--- 60 80 100 120 Lysolecithin/Amylose (w/w)
1EMPERATURE ( o C)
Fig. 2-Enthelpies of melting of amylose-lysolecithin complexes as
Fig. l-Thermogran,s .of HMW and LMW amyloses and amylopec- a function of lysolecithin:amylose. LMW amylose with egg lysoleci-
fin with egg lysolel:ithin, obtained on first heating (I) and after thin: first heating, 0; second heating, 0; third heating, 0. LMWamy-
cooling and reheatir 0 (IN. Weight of amylose or amylopectin lmgl lose with synthetic iysolecithin: first heating, a; second heating, X.
and weight ratio of lysolecithin to amylose or amylopectin: HMW HMW amylose with egg lysolecithin: first heating, A; second heating
amylose, 0.86 and 0 51; LMWamylose, 1.01 and 0.38;amylopectin. A; third heating 4 HMW amylosa with synthetic lysolecithin: first
1.35 and 0.08. heating. q ; second heating, n .
vent, either because of crystallinity or branching, then com- lysolecithm gives the same enthalpy as with egg lysolecithii.
plex formation with Lipid might be restricted. The X-ray A third heating did not change the enthalpy. Therefore,
diffraction measurements of the dry amyloses showed a the enthslpy obtained on reheating represents maximum
clear pattern for the HMW amylose and a diffuse pattern incorporation of lysolecithin into the starch. This enthalpy
for the LMW amylose. This indicates that the HMW amy- was used to estimate the amylose content of the starch.
lose has a higher degree of crystallinity than the LMW
amylose. Since the LMW amylose appears to be fully dis- Calculation of amylose content
persible in the solvent, and since the chain length of 300 To determine amylose content, the following formula
glucose units is not so short that end effects in formation was used:
of the complex with lipid would be significant, the en- enthalpy (Cal/g) for starch
thalpy of melting the LMW amylose-lysolecithm complex % amylose = 100 x
was selected as the standard enthalpy for maximum incor- 5.9 d/g
poration of lysolecithin into amylose. This enthalpy change,
5.87 f 0.1 Cal/g of amylose, was then used to estimate the
amylose content of starches.
When synthetic lysolecithin was added to amyloses in
place of egg lysolecithin, endotherms similar to those ob-
tained with egg lysolecithin were obtained. The peak tem-
peratures for the first heating were 103.6 and 103.3C,
and those for the second heating, 105.4 and 105.9C, for
HMW and LMW amyloses, respectively. Synthetic lysoleci-
thin gives a peak temperature about lC lower than that for
egg lysolecithin. However, the enthalpies of melting ob-
tained with synthetic lysolecithin were the same as those
obtained with egg lysolecithin.
Stoichiometry of the complex
The plot of enthalpy of melting of the amylose-lysoleci-
thin complex as a function of lysolecithin/amylose ratio,
Figure 2, shows a characteristic saturation plateau for bmd-
ing. For the re-heating data, the rising portion of the plot
(at low lysolecithin/amylose ratios), when extended linearly
to the plateau, indicates that binding equivalence is attained
at a lysolecithin/amylose ratio of 0.14, with an estimated
error of kO.02. This suggeststhat amylose can bind at most 0.1 0.2 0.3 0.4 0.5
about l/7 of its weight of lysolecithin.
The enthalpy of melting of the amylose-lysolecithin Lysolecithin/Potato starch (w/w)
complex of potato starch was determined as a function of
lysolecithin/starch ratio (Fig. 3). As with amylose, the Fig. 3-Enthalpies of melting of potato starch-lysolecithin complex,
enthalpy obtained on reheating was higher than that on as a function of lysolecithin:starch. Egg lysolecithin: first heating,
first heating. The enthalpies were constant at lysolecithin/ o; second heating, a; third heating, Q. Synthetic lysolecithin: first
starch ratios above about 0.05. The starch with synthetic heating A; second heating, 4
I I I , 1 I I , -r I I I , , , , ,
MIZE (N)
A 8
OlAlO
APIOCA
\
EAN
-xl
lAXY MAIZE 1
rAPlO> r\
.IMA BEAN 1 \/ /
LMVLOMAIZE
starches (Fig. 5). Defatting did not affect the amylose con- is formed exothermically during gelatinization (Kugimiya
tent of these starches, as determined calorimetrically et al., 1980). Quantitative calculations can be made by
(Table 1). comparing the enthalpy of melting of the complex
The amylose content of maize and wheat starches, as [AHi,,( Table 11 with the difference between the
calculated from the size of the endotherms of the lysoleci- enthalpies of gelatinization without [AH(G)] and with
thin-amylose complex, were significantly higher than amy- [AH,,(G)] lysolecithin (Table 2). Such calculations show
lose contents previously reported (Table 3), which are that within experimental error, for the first heating, the
based on iodine affinity. The reasons why these starches magnitude of the exotherm for formation of the complex
showed higher amylose contents is not clear. It is possible equals the magnitude of the endotherm for melting of the
that these starches contain an intermediate component or complex.
an amylopectin which is able to bind some lysolecithin A second heating of the starch-lysolecithin samples does
but is not able to bind iodine. Alternatively, and we think not give a gelatinization endotherm, but only an endotherm
this is less likely, the enthalpy of melting of the amylose- for melting of the amylose-lysolecithin complex. While the
lysolecithin complex formed by these starches may be
abnormally high.
Fractionation of maize and wheat starches gives a higher Table 1-Enthalpies of melting of starch-lysolecithin complexes and
content of amylose than obtained by iodine affinity. Whist- estimation of amylose contents
ler and Hilbert (1945) reported a 29-31% yield of crude
amylose from maize starch when 2-nitropropane was used Sequence Peak AHlysffNb Amylose
to form the complex with amylose. The yield from wheat of Temperatureb content
starch, using 1-nitropropane, was 31-32%. Lansky et al. Starch heatinga CC) (call91 (96)
(1949) reported that Pentasol gave a 28-29% yield of a
Potato I 104.9 f 0.2 1.22 f 0.03
linear component from maize starch, although n-butyl II 106.9 f 0.2 1.42 f 0.06 24
alcohol gave 22-23%. The amylose contents obtained by III 106.7 f 0.0 1.47 + 0.00
precipitating water-insoluble complexes thus seem higher
than those obtained by iodine affinity. Purely on entropy Tapioca I 103.6 f 0.1 0.79 f 0.15
considerations, the stability of complexes formed between II 106.8 f 0.1 1.14 i 0.06 19
short chains and the relatively large lipids or similar or-
ganic molecules could be significantly greater than the sta- Maize If 103.5 +_0.4 1 A4 i 0.02
bility of short-chain complexes with the smaller iodine (nondefatted) I 107.4 + 0.6 1.94 f 0.07 33
molecules. Accordingly, the discrepancy in the amylose Maize I 103.4 f 0.2 1.51 f 0.00
content of maize and wheat starches may be due to the fdefatted) II 107.2 f 0.3 1.96 f 0.05 33
presence of a fraction with properties intermediate between
amylose and amylopectin, or of an amylose fraction of Wheat I 103.4 + 0.2 1.36 + 0.04
short chain length. If so, these fractions must form com- (nondefatted) II 107.1 f 0.3 2.16 f 0.02 37
plexes with lysolecithin to a greater extent than with iodine.
The present results, and the fractionation experiments Wheat I 103.4 5 0.2 1.37 f 0.10
referred to above, suggest that maize and wheat starches (defatted) II 107.3 f 0.3 2.16 f 0.03 37
may have a considerable amount of such intermediate frac-
tions. It has been recommended that the amylose con- Lima bean I 104.0 1.39
II 107.2 2.01 34
tent of starches be expressed merely in terms of iodine
affinity, with no reference to percentage of amylose present Wrinkled pea I 104.8 f 0.2 2.32 + 0.03
(Lansky et al., 1949). II 107.5 + 0.6 3.80 * 0.06 65
Other starches
Amylomaize I 106.0 +_0.3 2.30 f 0.12
The amylose contents of most of the other starches, as II 107.0 f 0.1 3.83 f 0.08 66
determined by this calorimetric method, are in good agree-
ment with the reported amylose contents (Table 1 and 3). Waxy maize I 103.3 f 0.0 0.03 f 0.00
Amylomaize was found to be 66% amylose by the calori- II 107.8 f 0.3 0.04 f 0.00 0.6
metric method. Banks et al. (1974) have reported that E I, II, and III designate first, second and third heatlng.
iodine-binding measurements yield a higher estimate of the Mean and standard deviation for triplicates, except for lima bean.
amount of amylose present in amylomaize starches (79%)
than does butanol-complex formation (63%) and that the
discrepancy between these independent measurements of Table 2-Enthalpies of starch gelatinization with and without
amylose content represents material, abnormal amylopec- lysolecithin
tin, which is neither amylose nor amylopectin, in the con-
Without Lysolecithin With Lysolecithin
ventional sense, Their explanation for this is that amylo-
maize amylopectin is heterogeneous, and is composed of a TQa AH(GIa AHI (G)
high molecular weight material indistinguishable from Starch (C) (cal/g) (2, kaVd
normal amylopectin and a low molecular weight composed,
Potato 64.8 f 0.1 4.43 f 0.11 64.7 f 0.0 2.97 f 0.13
at least partially, of amylose of relatively short (<lOO
glucose units) chain length. If the discrepancy in amylose Tapioca 71.1 f 0.1 3.14 f 0.06 70.6 f 0.0 2.01 f 0.16
content obtained by the different methods arises from the
presence of intermediate material, the intermediate material Maize
of maize and wheat starches must be different from that of (defatted) 70.0 f 0.4 2.89 * 0.17 69.8 f 0.2 1.59 f 0.26
amylomaize starch.
Lima bean 80.6 4.26 79.9 2.91
DISCUSSION
THE ENTHALPIES of the gelatinization transition of Waxy maize 73.0 f 0.2 3.82 + 0.09 73.0 f 0.0 3.51 f 0.17
starches with added lysolecithin are smaller than without a Mean and standard devlatlon for triplicate determinations, except
lysolecithin. This suggests that all or most of the complex for lima bean.
high temperature endotherm obtained on first heating has a method is still to be explained. However, high amylose
single, broad peak, the endotherm obtained on reheating is contents, in themselves, do not present a difficulty if the
a peak with a le;.ding shoulder. The asymmetry of the calorimetric method is used to compare one variety of
endotherm obtained on reheating appears to be related to wheat or maize starch with another. For the other starches,
the state of subdivision of the amylose-lipid complex. The the amylose content obtained by the calorimetric method
sharp peak at 107C appears to result from melting of crys- agrees well with the amylose content obtained by the io-
talline complex. The leading shoulder, and the endotherm dine-binding method, so that there should be no difficulty
obtained at 104-l 05C on first heating, appears to result in using the calorimetric method for these starches.
from melting of ei:her noncrystalline complex, or complex
with a much 1ow:r degree of order. The proportion of REFERENCES
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Adkins. G.K. and Greenwood, C.T. 1969. Studies on starches of
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-Continued on page 777