Accepted Manuscript

Title: Interrelation between thermal behaviour and pasting

properties of malted rice using multivariate analysis

Authors: Dipsikha Kalita, Suvendu Bhattacharya, Brijesh

Srivastava

PII: S0040-6031(18)30416-7
DOI: https://doi.org/10.1016/j.tca.2018.10.008
Reference: TCA 78114

To appear in: Thermochimica Acta

Received date: 20-6-2018

Revised date: 8-10-2018
Accepted date: 12-10-2018

Please cite this article as: Kalita D, Bhattacharya S, Srivastava B, Interrelation between
thermal behaviour and pasting properties of malted rice using multivariate analysis,
Thermochimica Acta (2018), https://doi.org/10.1016/j.tca.2018.10.008

This is a PDF file of an unedited manuscript that has been accepted for publication.
As a service to our customers we are providing this early version of the manuscript.
The manuscript will undergo copyediting, typesetting, and review of the resulting proof
before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that
apply to the journal pertain.
Interrelation between thermal behaviour and pasting properties of malted
rice using multivariate analysis

Dipsikha Kalita, Suvendu Bhattacharya and Brijesh Srivastava*

Department of Food Engineering and Technology
School of Engineering, Tezpur University,
Assam 784028, India

T
R IP
SC
Running title: Pasting and thermal behaviour of malted rice

U
N
A

* Address for the corresponding author

M

Dr. Brijesh Srivastava

Associate Professor
Department of Food Engineering & Technology
D

School of Engineering, Tezpur University, Assam-784028, India

Tel: +91-3712-27-5712
TE

E-mail: brijesh@tezu.ernet.in
EP

Highlights
 Thermal behaviour malted rice was affected by germination time
CC

 A biphasic pasting curve was observed in malted RLA

 Malted RNA showed higher extent of decrease in pasting properties

A

 PCA revealed strong correlation between pasting and thermal properties

Abstract

The interrelation between thermal behaviour and pasting properties in malted rice, affected by time/ temperature of

germination, was established using multivariate analysis. Two paddy cultivars, low (RLA) and normal amylose

(RNA)were germinated at 30 and 35°C for120h. Thermal behaviour from DSC plots (onset, peak and conclusion

temperatures) of rice increased with germination time; RNA showed a higher decrease in pasting properties compared

to RLA. A biphasic pasting curve for malted RLA indicated amylose-lipid complex formation during heating. This

complex resulted in increasing trends of all DSC parameters. The principal component analysis biplots of RLA at both

T
IP
temperatures showed that the position of amylose was loaded away from the vicinity of pasting and thermal properties;

in RNA, amylose content was in the neighbourhood of these properties. Inter-relationship between the pasting and

R
thermal behaviour of malted rice existed; the information might help for developing malted rice-based weaning and

SC
functional foods.

U
N
Keywords: Malted rice; viscoamylography; thermal behaviour; kinetics; inter-relations; principal component analysis
A
M
D
TE
EP
CC
A
1. Introduction
Rice is the most widely consumed staple food of the world population among which Asian countries are the major
consumers of rice and rice products. Rice is commonly consumed after cooking in boiling water. Several products
including ethnic foods are also prepared using rice flour as the main ingredient though other raw materials like pulses
and vegetables are commonly incorporated [1]. The worth mentioning the advantages of rice is that it does not contain
gluten and is the least allergenic grain [2]. These advantages make the rice-based formulations a reasonable choice for
preparing gluten-free products for people suffering from celiac disease. It is a digestive tract disorder for a human who
must avoid gluten-containing foods throughout their lifespan [3]. Thus, a need exists to develop gluten-free products
which can match the product characteristics of popular wheat-based products. These considerations are thus helpful in

T
developing specialty foods.

IP
Several cereals, millets and legumes are subjected to the germination process and the malted flour is

R
incorporated to prepare health-benefitting foods. Malting of grains is done by steeping, germination, kilning/drying and

SC
grinding. Malting or controlled germination is a traditional process of modification of starch [4]. Germination
commences with the rapid imbibition of water by the dry seeds which result in the hydration of all the matrices and cell
contents. Gibberellic acid (GA) plays an important role during the germination process as it stimulates the synthesis of

U
hydrolytic enzymes such as α- and β-amylases which otherwise are not active in the dry raw seeds. These enzymes
N
induce partial degradation of the reserved compounds in the grain especially the starch into simpler compounds like the
A
various reducing sugars [5].
M

The modification of starch by the process of germination is expected to modify the pasting property and
thermal behaviour which play important roles during processing. The characterization of cereal biopolymers by
D

thermal analysis is important in apprehending the functionality of starch on the structural and molecular levels [6]. The
TE

pasting profile of rice flour is another key parameter which is associated with starch gelatinization indicating the
cooking behaviour and possibly correlates to eating quality. Limpisut et al. [7] have indicated that the peak viscosity
EP

correlates positively with consistency and hardness of cooked rice whereas a negative relationship exists with
stickiness. The process of germination brings about various modifications due to decomposition of high molecular
CC

weight compounds. An increase in reducing sugars occurs while the viscosity of the malted rice slurry decreases due to
the amylolytic effect on starch [8]. The decrease in viscosity is also reported by Ayernor et al. [5] for samples
germinated for 9-days at 32°C. However, the understanding of the effect of amylose and amylopectin ratio on the
A

pasting properties and thermal behaviour of malted rice at different germination conditions are still inadequate.
Moreover, the correlation among pasting properties and thermal behaviour of malted rice using a statistical
approach such as multivariate analysis is not reported and requires detailed investigations. Chemometrics (multivariate
data analysis) can be described as the interaction of certain mathematical and statistical methods in chemical
measurement processes [9]. Principle component analysis (PCA) is one of the most used chemometric methods which
does not require prior knowledge of the sample and can provide patterns, groupings, detection of outliers, etc. [10]. It
mainly analyses the data which is described by several dependent variables and express the information as a set of set
of new orthogonal variables called principal components. It thereafter represents the pattern of similarity of the
observations and the variables by displaying them as points in maps [11].

Thereby, the objectives of the present study are (a) to determine thermal behaviour and pasting properties of
malted rice under different germination conditions, and (b) to establish the interrelations among/between the indices of
thermal behaviour and pasting properties using multivariate analysis. The results are expected to provide an overview
of the thermal behaviour and pasting properties of malted rice flour which is affected by time course germination
process. This information shall assist in the development of malted rice flour based weaning food formula, and bakery
and confectionery products.

T
IP
2. Methods

R
SC
2.1. Materials
Two paddy varieties having different amylose and amylopectin content viz. Chokuwa as low amylose rice (amylose:
12.5±0.1, amylopectin: 62.1±0.9, fat: 0.7±0.1 and protein: 3.5±0.2%) (RLA) and Aijong rice as the normal amylose

U
rice (amylose: 20.2±0.8, amylopectin: 20.2±0.7, fat: 0.8±0.1 and protein: 3.8±0.1%) (RNA) were selected for the
N
present study. The paddies were procured from the cultivars of Nagaon, Assam, India and had a moisture content of
A
about 12%.
M

2.2.1. Malting of paddy

Malting of paddy was performed according to the method described elsewhere [12, 13]. In brief, the 24 h soaked paddy
D

was germinated at 30˚C and 35˚C for 5-days, and samples were drawn at an interval of 24 h. Later, each sample was
TE

dried in a hot air dryer at 50˚C for 24 h followed by dehusking and grinding. The malt powder samples thus obtained
were packed and stored in refrigeration until further analysis. The paddy samples germinated at 30 and 35°C were
EP

coded as RLA30 and RLA35 for low amylose, and RNA30 and RNA35 for normal amylose, respectively. Amylose
content of the rice was determined in triplicates by the method suggested by Labuschagne et al. [14]. Amylopectin
CC

content was calculated by subtracting amylose content from the total starch content as detailed by Wu et al. [15].

2.2.2. Thermal behaviour

A

The thermal behaviour of samples was measured employing a differential scanning calorimeter (DSC) (214, Polyma,
NETZSCH, Germany). The samples were saturated with distilled water (1:3) and placed in an aluminum pan for 24 h
in a desiccator. The pan was sealed with a lid for examination under DSC. The reference employed for the
measurement was an empty pan with the lid. The samples were heated from 20 to 150°C at a heating rate of 10 K/min.
The onset (TO), peak (TP) and completion (TC) temperatures were computed from the thermogram.

2.2.3. Pasting properties

The pasting properties were determined with a rapid visco-analyser, RVA (Starch master 2, Newport Scientific,
Warriewood, Australia). The samples (3.2 g, 14 g/100 g moisture basis) were mixed with 25.3 ml of distilled water in
canisters. The slurry was mixed at 960 rpm for the first 10 s at 29oC followed by 160 rpm for the rest of the run. The
steps for heating and cooling cycles were followed as discussed below:
Step-I: heated to 40⁰ C for 1 min;
Step-II: heated up to 50⁰ C within 3 min;
Step-III: heated to 95°C within 3.4 min;
Step-IV: holding at 95⁰ C for 3 min; Step-V, and subsequently cooled to 50°C.
The parameters evaluated were the pasting temperature (PT), peak viscosity (PV), hot paste viscosity (HPV),

T
breakdown viscosity BDV), setback viscosity and cold paste viscosity (CPV). The breakdown viscosity (BDV) was

IP
calculated as the difference between IPV and HPV, and the setback viscosity (SBV) was equal to the difference of
CPV and HPV.

R
SC
2.2.4. Kinetics of pasting and thermal properties
Changes in pasting properties and thermal behaviour with respect to germination time were modeled with the zero, first
and second order kinetics (Eqs. 1, 2 and 3, respectively) [16], and rational function (Eq. 4), respectively. The rate

U
constants (k1, k2 and k3) for the changes were estimated from the slope of the plots against time arising from Eqs. 1, 2
N
and 3. The rational function employed for the thermal behaviour could be defined as the ratio of two polynomial
A
functions. If P(x) and Q(x) were the polynomials, then a function of the form R(x) =P(x)/Q(x) was a rational function
where Q(x) ≠ 0. Equation (4) was a combination of linear and quadratic functions of time t which were represented by
M

the numerator and denominator, respectively.

D

At = k 0 t + A0 Eq. (1)
TE

ln At = k1 t + ln A0 Eq. (2)
EP

1 1
CC

= k2t + Eq. (3)

At A0
Here, At and A0 were the values at the beginning and after the treatment time t. The model with the maximum R2 was
A

selected to represent the dependency of the input variables.

𝑎+𝑏𝑡
𝑌 = 1+𝑐𝑡+𝑑𝑡 2 Eq.(4)

Here, Y was equal to TO, TP and TC, and a, b, c, d were the model parameters. The estimation of the parameters of Eq.
(1) was performed by employing the tool of MATLAB, version 2014a.

2.3. Multivariate (chemometric) analysis

PCA helps in analyzing the relationship between the original variables, the way they correlate with each other, and
their influence in determining the new coordinate system. PCA was performed [17] to inter-relate pasting and thermal
properties, and independent variables. The variables were selected as continuous variables. The algorithm employed
was the nonlinear iterative partial least squares (NIPALS). NIPALS is a well-established iterative technique widely
used in building PCA. The optimal number of components were analyzed by the method of cross-validation. A PCA
biplot plot was generated to study the interrelation between the variables.

2.4. Pearson correlation and statistical analysis

The data were analyzed by employing the analysis of variance (ANOVA) using IBM Statistical Package for the Social

T
Sciences (SPSS, version 20). Duncan's multiple range test (DMRT) was performed to identify the terms having
significant differences at p≤0.05. Pearson correlation analysis was also conducted to determine the relationships

IP
between/among the pasting and thermal properties

R
SC
3. Results and discussion
3.1. Role of amylose and amylopectin
Starch is the major component of rice and acts as the main source of energy during germination for the growing

U
embryo. The starch content decreased markedly in both RLA and RNA during the germination (Table 1) due to the
N
action of hydrolytic enzymes. Moreover, RNA showed higher starch degradation as compared to that of RLA which
A
might be due to the presence of higher content of amylose in RNA as discussed in our earlier study [23]. Amylose and
amylopectin are two major components of rice and their changes during germination process are shown in Table 1. An
M

increasing trend of amylose content was observed in RLA30 and RLA35samples while the amylopectin content
decreased significantly (p≤0.05). It might be due to the degradation of branched structure of amylopectin by hydrolytic
D

enzymes such that amylose content increased. Atichokudomchai et al. [18] reported that amylase caused a rapid
TE

decrease in the molecular weight of amylose and amylopectin. Germination temperature also had a significant effect on
the amylose and amylopectin contents of both RLA and RNA. Wu et al. (2013) reported a decrease in the amylose and
EP

amylopectin contents during paddy germination for 5-days; the extent of decrease in amylopectin was greater than
amylose content. The breakdown of amylose and amylopectin resulted in an increase of the reducing sugar content
CC

during the germination process (Table 1). This might also be attributed to the phenomenon of starch degradation due to
hydrolytic enzymes (α and β amylases) [23].
A

3.2 Thermal behaviour

The thermal behaviour of samples obtained from the DSC plots in terms of the onset temperature (TO), peak
temperature (TP) and completion temperature (TC) are shown in Table 2 and Fig. 1. These indices were affected by the
amylose-amylopectin content, germination time and temperature. The TO reflects the initiation of gelatinization
phenomenon in starch samples, and in the present study, the TO values of non-germinated samples were between 64.1
and 65.7°C. On the other hand, gelatinization temperatures (TO and TP) of the native RNA were higher compared to
RLA. This might be attributed to the variation in the amylose and amylopectin contents. In high amylose rice, the chain
association between amylose decreases the hydration of the amorphous region of the starch granules and thereby
retards swelling and gelatinization [19]. Park et al. [20] also reported similar observations where non-waxy rice showed
higher TO and TP as compared to waxy rice which was attributed to the presence of the crystalline region in the waxy
rice which required less energy for melting.

The onset temperature (TO) and peak temperature (TP) of malted RLA and RNA samples increased till 24 h of
germination. This might be attributed to the in-situ formation of the starch-lipid complex. It is reported that in excess
water, as in the case of DSC, a total dispersion of amylose molecules in aqueous phase occurs which makes them
readily available for complexing with the dispersed liquids. Moreover, crystalline amylose-lipid complexes can be
obtained especially when crystallization occurs at a high temperature. Similarly, Wokadala et al. [21] showed high TO,

T
TP and TC values after α-amylase hydrolysis in teff and maize starch. According to Bail et al. [22], cereals with high

IP
amylose content contained a greater amount of lipid (starch-lipid complexes) might result in a high final gelatinization
temperature. However, TO and TP decreased between 24 and 120 h of germination which might be due to the

R
breakdown of glycosidic linkages in amylose leading to the formation of smaller molecular weight compounds such as

SC
glucose, maltose and maltodextrins. The structural changes in the glycosidic linkages during germination were also
observed from FTIR and Raman spectroscopy in our earlier studies [13]. Thus, only a small amount of amylose was
leached that could not complex with lipids leading to a decrease in the T O and TP values. The TO of native RLA and

U
RNA was not statistically different (p≤0.05), but malted RNA showed lower values than malted RLA as germination
N
progressed; the effect of germination temperature was not statistically different (p≤0.05) for RLA, but TO was lower
A
for RNA35 compared to RNA30 (Appendix Table 2). This might be due to the higher starch degradation by various
hydrolytic enzymes in malted RNA which was also observed in our earlier study [23]. The end set or conclusion
M

temperature (TC) reflects the thermal status of the sample when the crystalline structure gets transformed into an
amorphous phase. The TC followed the same trend as TO and TP in malted RLA and RNA samples and thereafter
D

decreased up to 120 h of germination.

TE

3.3 Pasting properties

EP

A rapid visco-analyser (RVA) is a tool to monitor the changes during cooking. The pasting behaviour of malted RLA
and RNA varieties is shown in Fig. 2 and Table 3.
CC

3.3.1. Pasting temperature

The pasting temperature (PT) indicates the minimum temperature required to gelatinize/cook a starch-rich sample.
During gelatinization, water is absorbed by the sample which involves granular swelling, melting of the crystalline
A

zone of starch, exudation of the molecular components from the granular portion and eventually total distribution of the
granules. The native RLA showed a lower PT compared to RNA (Fig. 2) which might be due to a variation in the
amylose content. Jang et al. [19] reported that higher amylose content of rice starch had higher PT. The RLA and RNA
showed a decrease in PT as germination time increased indicating a faster swelling and gelatinization of the germinated
starch granules. Similar results were also observed by Musa et al. [24] for germinated brown rice which swelled faster
than the brown rice and white rice. Germination caused enzymatic hydrolysis of starch which felicitates the swelling of
granules. It was observed that the PT of RVA corresponded to the TO values of the malted rice samples. However, the
TO values of non-germinated samples (65.7-66.0°C) were lower than that of PT values (71.0-75.0°C). Woo et al. [25]
reported the TO of starches from 10 rice cultivars to be between 57.9 and 64.4°C which was lower than those obtained
in the present investigation. These differences might be attributed to the differences in the principle of measurement.
The ANOVA (Appendix Table 1) showed that the independent variables amylose/amylopectin content and germination
time had significant effects though germination temperature did not have a significant effect. However, the interaction
between independent variables did not have a significant effect (p≤0.05) on PT.

3.3.2. Peak viscosity

Peak viscosity (PV) of a starch-rich sample reflects the water binding capacity; the magnitude also indicates the energy
required for mixing and transferring of the viscous sample [26]. The native RNA exhibited higher PV than native RLA

T
which might be attributed to the higher amylose content in RNA. A similar result was also reported by Noda et al. [27].

IP
The ANOVA indicated the significant effect of amylose-amylopectin content, time and temperature on PV.

R
Two peak viscosities were observed in RLA30 and RLA35 (Fig. 2). A steep rise in viscosity was noticed from

SC
the pasting point and the first peak appeared at about 4 min which was followed by a decrease in viscosity. Unlike
RNA, a second PV was observed after a few minutes followed by a decrease in viscosity before setback viscosity. The
biphasic peak observed for malted RLA samples was possibly due to the formation of the starch-lipid complex. It was

U
reported that in cereal starches, lipids are present both on the surface and inside the granule, and they may be in the free
N
or bound state to starch components [22]. As amylopectin content is higher in RLA, its outer branches might assist in
A
the formation of a helical inclusion complex with lipid [28]. However, it was observed that the complex did not survive
for a long time as dissociation occurs due to the shearing action in the viscoamylograph in addition to the heating
M

process which eventually caused a fall in viscosity. The magnitude of the second peak was higher than that of the first
peak viscosity, but both the peaks decreased significantly during the phase of germination due to the breakdown of
D

amylopectin. A similar biphasic pasting curve due to the formation of amylose-lipid inclusion complexes was reported
TE

by Nelles et al. [29] for maize starch during prolonged holding time when subjected to viscoamylographic testing.
EP

Germination process hydrolyzes starch to smaller molecules by the secreted hydrolytic enzymes. The α-
amylase being an endo-amylase cleaves the α-1, 4glycosidic bonds from the inner regions of starch which decreases
CC

viscosity. On the other hand, exo-amylases also known as saccharifying enzymes, hydrolyze starch from the non-
reducing ends to yield maltose and glucose [23]. However, both α- and β-amylases cannot cleave the α-1→6 glycosidic
linkages which produce α- and β-dextrins; these dextrins of sufficient degree of polymerization (DP) might have
A

complexed with endogenous lipids present in starch during the process of gelatinization through ionic or hydrogen
bonding to exhibit the second PV [30]. Wu et al. [15] reported a similar decrease in PV during the germination of
brown rice. It was attributed to a decrease in starch content by the amylase activity. In the case of RNA samples, a
higher degradation of starch was observed; conversion to simple reducing sugars was expected by various hydrolytic
enzymes without amylose-lipid complex formation. The change in PV during germination might also be attributed to
the loss of crystallinity of starch granules which was reported in our earlier study [23].

3.3.3. Hot paste viscosity

The peak viscosity is followed by the hot peak (paste) viscosity (HPV), and it is the viscosity of the system at the end
of cooking/heating time at 95oC. The HPV primarily indicates the fragility of the swollen grains towards shear
treatment. During the holding period at 95°C, disruption of granules occurs due to shearing offered by the rotating
paddle and bowl. This phenomenon resulted in the formation of a viscous mass/paste; the cooked starch paste consisted
of a continuous phase of solubilized starch fraction and a discontinuous phase of granule remnants (outer portion of the
granule) [31]. The germination time had a significant effect on HPV compared to amylose-amylopectin content and
temperature (Appendix Table 1).

The native RLA showed higher HPV compared to RNA (Fig. 2). It is possible that a variation in the amylose

T
and amylopectin contents had a significant effect on HPV. Amylopectin is usually responsible for swelling of starch

IP
granules whereas amylose and lipids impede granular swelling and maintain the integrity of the swollen granules
during pasting [19]. Therefore, higher swelling of RLA granules increased its susceptibility to shear damage resulting

R
in high HPV. Comparable results were also reported by Ashogbon and Akintayo [32] who studied the pasting

SC
properties of starch from different cultivars of rice from Nigeria.

The HPV of RLA and RNA samples decreased with germination time due to the action of amylolytic enzymes;

U
the amylose-amylopectin content showed a significant effect. RNA showed a higher decrease of HPV compared to
N
RLA at both germination temperatures which might be due to its high amylopectin content that affected the swelling of
A
granules and their disintegration.
M

3.3.4. Breakdown viscosity

Breakdown viscosity (BDV) measures the susceptibility of cooked starch towards disintegration during continuous
D

stirring and heating; it is calculated as the difference between the PV and HPV. During heating in the viscoamylograph,
TE

the swelled granules had a less space to expand further, and the Brownian movement made the swollen granules to
collide with each other, and thus disintegrated to a great extent. However, the breakdown depended on the rigidity and
EP

structural integrity of the swollen granules.

CC

The BDV of native RNA was higher than that of RLA (Fig. 2). The high breakdown indicated the sensitivity
towards heat and stress during cooking [33]. This result was not in agreement with the previous studies which had
suggested a less breakdown of RNA due to the presence of amylose that hindered granule swelling [19]. Park et al. [20]
A

indicated that although RNA was resistant to gelatinization, they broke down easily once they had been gelatinized.

The BPV for both RLA and RNA decreased drastically after 24 h of germination whereas it further decreased at
a slower rate. However, a significant effect (p≤0.05) was observed between amylose-amylopectin content, time and
temperature (Appendix Table 1). The malted RNA showed a higher decrease (about 20%) compared to malted RLA
(about 10%) at both germination temperatures.

3.3.5. Setback viscosity

Setback viscosity (SBV) or retrogradation index refers to the process of cooling of a heated starch dispersion in which
the exuded amylose molecules re-associate and the swollen starch granules are united to provide an ordered structure
resulting in an increase of the system viscosity. This phenomenon is also known as the recrystallization process of
amylopectin and amylose during which the formation and aggregation of double helices take place. A high SBV
implies a great extent of re-crystallization of gelatinized starch during the phase of cooling of starch-rich samples [33].
Higher SBV values were observed for native RLA compared to RNA. Ashogbon and Akintayo [32] and Kong et al.
[34] reported that the amylose component in the starch had the higher retrogradation tendencies. The higher SBV of
native RLA sample might be attributed to the longer chain length of amylopectin which helped the development of
retrograded fractions.

T
IP
The SBV of malted RLA and RNA samples increased during the initial 24 h of germination. This was expected
due to the hydrolysis of amylose and amylopectin molecules which matched the earlier results of Musa et al. [24] for

R
germinated brown rice. This phenomenon is related to chain length and contents of amylose and amylopectin in starch.

SC
The interaction between amylose-amylopectin content, time and temperature showed a significant difference in setback
viscosity (Appendix Table 1). The RNA30 and RNA35 showed the higher decrease compared to RLA30 and
RLA35samples which might be attributed to the greater extent of hydrolysis of starch granules by amylolytic enzymes.

U
N
3.3.6. Final or cold paste viscosity
A
Cold paste viscosity (CPV) or final viscosity is the viscosity of the system at the end of the cooling phase at 50°C; the
increase in viscosity is due to retrogradation or re-association characteristic of amylose due to a decrease in the system
M

temperature. The viscosity measured at 50°C might be linked to the eating quality of foods. The native RNA showed a
higher CPV compared to RLA which might be attributed to the linear chain structure of amylose to aid the formation
D

of hydrogen bonds resulting in the development of firm gels [34].

TE

The CPV of the malted RLA and RNA for both temperatures decreased as germination time increased. RNA30
EP

and RNA35 exhibited a higher decrease in CPV compared to RLA30 and RLA35. This might be attributed to the
presence of higher amylose content in RNA and the linear structure was possibly gets degraded quickly by the
CC

hydrolytic enzymes in the malted rice. Thus, the formation of the hydrogen bond was affected during the cooling
process resulting in a decrease of CPV. The interaction between amylose-amylopectin content, time and temperature
showed significant effects on CPV (Appendix Table 1). The decrease in CPV with germination time was mainly due to
A

amylase activity; the enzyme activity increased with germination time to enhance the breakdown of starch into small
fragments and exhibited a decline in CPV. The pasting property had been reported to be affected by the presence of
proteins and lipids, and the formation of amylose-lipid complexes [35].

3.4. Kinetics of pasting properties and thermal behaviour

The second-order kinetic model was best suited (R2 between 0.721 and 0.970; significant at p≤0.01) for the pasting
properties followed by the first-order model (R2 between 0.698 and 0.99) though the zero-order model was unsuitable
(R2 between 0.115 and 0.862); the model parameters are shown in Table 4 and Appendix. The root means square error
(RMSE) values also followed the same trend. Similarly, the rational function (Eq. 4) employed for the thermal
properties (Table 4) showed high R2 ranging from 0.95 to 0.99 (significant at p<0.05). It is thus inferred that the model
second-order kinetic was best suited for the pasting properties while the complex rational model was applicable for the
thermal properties.

3.5. Inter-relationships between pasting and thermal properties using multivariate analysis
The inter-relation between thermal behaviour and pasting properties was established based on the PCA (Fig. 3-4). The
PCA is a powerful tool for visualization of the inter-relationships between/among the independent variables and
dependent parameters that may be used to explain the maximal variance of the data [36]. The principal components

T
(PC), as biplots, which explained the variance in the pasting and thermal properties of RLA and RNA are presented in

IP
Fig. 3 and 4, respectively. The biplot of pasting properties and thermal behaviour of RLA30 showed that the first (PC1)
and second (PC2) principal components accounted for 71.7% and 25.5% of the variance, respectively, explaining a

R
total of 97.2% of the variation. The pasting properties (PV2, SBV, CPV, HPD and BDV) could be grouped together

SC
and thus they behaved in a similar manner. This group was observed in the vicinity of PT and amylopectin content
exhibiting a high correlation among them. Both the group parameters showed a significant decrease with the
germination process due to the breakdown of starch by amylolytic enzymes. On the other hand, PV1 and thermal

U
properties (TO, TP and TC) were loaded away from these two groups and was expected to have an inverse relationship
N
between them. The amylose content was loaded on the opposite quadrant of that of amylopectin meaning an inverse
A
relationship. In RLA30, amylose content increased significantly due to the breakdown of the crystalline region to the
amorphous region. Thereby, amylose content showed a negative correlation with the thermal and pasting properties.
M

Woo et al. [25] had also reported that no correlation was observed between amylose content and any thermal parameter
measured by DSC. Their results were also in accordance with earlier reports that showed a weak correlation between
D

amylose content and thermal characteristics of starches [27].

TE

The PCA biplot of RLA35 showed that PC1 and PC2accounted for about 96% of the total variation. Here, the
EP

similar correlation among pasting properties, thermal properties and amylose and amylopectin contents was observed
as exhibited by RLA30.
CC

The biplot of pasting properties and thermal behaviour of RNA30 is shown in Fig.4. The PC1 and PC2jointly
explained 97.5% of the total variation. The pasting properties (PV, SBV, CPV, HVD and BDV) formed a group and
A

thus expected to behave in the same manner. This group was in the vicinity of PT, amylose and amylopectin contents
exhibiting higher correlation among them. These group parameters decreased during 120 h of germination due to the
action of amylolytic enzymes on starch. Significant linear correlations were observed between amylose content, lipid
content, gelatinization temperature and gelatinization enthalpies for rice starches [30]. On the other hand, TO, TP and
TC were close by and had an inverse relationship with the groups mentioned above. This was due to an increase in these
temperatures during the germination process.
 The biplot of pasting properties and thermal behaviour of RNA30explained 94.5% of the total variation. Similar
correlations were observed among pasting and thermal properties like that of RNA30. Ye et al. [37] worked on brown
flours obtained from four Indica rice subspecies with varying amylose contents, and their thermal and rheological
properties were examined. The results showed an increase in peak, final and setback viscosities (p<0.05) with an
increase in amylose content while the phase transition temperatures of brown rice flour (p<0.05) decreased.

3.6. Pearson correlation

The correlation coefficient matrix showed (Table 5) that the amylose content was negatively correlated to pasting and
thermal properties for RLA30 and RLA35. On the other hand, amylopectin was positively correlated with the pasting

T
properties but negatively correlated with the thermal behaviour. This was due to the breakdown of crystalline region of

IP
starch into the amorphous state by various hydrolytic enzymes. PT showed positive correlation with HPV, SBV and
CPV (r=.0.827, 0.868 and 0.842, respectively, p <0.05) for RLA30. PT decreased with germination time a while a high

R
gelatinization onset temperature was observed in 24 h of germination indicating resistance towards granule swelling; it

SC
later decreased with germination time. The first peak viscosity in RLA30 showed a positive correlation with TO
(r=0.94, p<0.01). In RLA30 and RLA35, PV2 positively correlated to HPV, BDV, SBV and CPV (p<0.01).

U
In RNA30, the amylose and amylopectin contents had higher positive correlations with pasting properties rather than
N
the thermal properties which indicated that amylose played an important role in pasting properties. The amylose and
A
amylopectin contents showed positive correlations with PT (r=0.896 and r=0.905, respectively, p<0.05). The PT
positively correlated with PV, HPV, BDV, SBV and CPV (p<0.05). Similarly, the PV exhibited positive correlations
M

with HPV, BDV, SBV and CPV (r=0.983, 0.998, 0.997 and 0.998, respectively, p<0.01). Similarly, RNA35 showed
that amylose and amylopectin contents had positive correlations with pasting and thermal properties. In RNA35, PV
D

showed higher positive correlations with HPV, BDV, SBV and CPV (p<0.01). Moreover, a positive correlation was
TE

observed between PT and TO but negatively correlated with TP and TC. On the other hand, T O, TP and TC showed
negative correlations with PV, HPV, BDV, SBV and CPV.
EP

4. Conclusions
CC

The thermal and pasting properties of two malted rice having different amylose content (RLA and RNA) were studied
during the time course germination process. The thermal behaviour of malted RLA and RNA samples were
significantly affected by the germination time, temperature, and amylose-amylopectin content. The values of TO, TP
A

and TC increased significantly for 24 h of germination and decreased thereafter. The germination time also had a
significant effect on pasting properties as they decreased with germination time. A biphasic pasting curve (two pasting
peak viscosities) was observed in malted RLA samples. The increase in thermal behaviour and the biphasic nature of
pasting curve may be due to the formation of the amylose-lipid complex during the heating process. Higher extent of
decrease in pasting properties implied the role of amylose-amylopectin content on the germination process for malted
RNA variety. The second-order kinetic model was best suited for the pasting properties and the change in the thermal
behaviour followed a rational function with respect to germination time. PCA biplots revealed that the amylose content
in RLA at both 30 and 35°C was loaded opposite to pasting and thermal properties which indicate negative correlation
among them. However, RNA at both germination temperature showed loading of amylose and amylopectin content
close to pasting and thermal properties. The correlation matrix showed a positive correlation between PT and TO in
both malted rice samples revealing the swelling of starch granules during hydrothermal treatment. PT also showed a
positive correlation with other pasting properties. A good correlation was also observed between amylose-amylopectin
content, pasting properties and thermal behaviour. Thus, this study showed the existence of an inter-relationship
between the pasting properties and thermal behaviour of malted rice as affected by the time course germination
process. The information might be useful for developing malted rice based weaning formulation and functional foods.

T
R IP
SC
U
N
A
M
D
TE
EP
CC
A
Acknowledgment
The authors acknowledge CSIR, New Delhi (Govt. of India) for awarding the fellowship and DST-FIST; NEQIP-
AICTE and UGC-SAP for providing financial support for infrastructure development of the Department of Food
Engineering and Technology, Tezpur University, India.

References

[1] Blandino, A., Al-Aseeri, M. E., Pandiella, S. S., Cantero, D., & Webb, C. (2003). Cereal-based fermented foods
and beverages. Food Research International, 36(6), 527-543.
[2] Rolfes, S. R., Pinna, K., & Whitney, E. (2014). Understanding normal and clinical nutrition. 9th ed. Belmont

T
Cengage Learning, Pp. 500–502.

IP
[3] de la Hera, E., Martinez, M. & Gomez, M. (2013). Influence of flour particle size on quality of gluten-free rice
bread. LWT-Food Science and Technology, 54, 199–206.

R
[4] Mohan, B. H., Malleshi, N. G., & Koseki, T. (2010). Physico-chemical characteristics and non-starch

SC
polysaccharide contents of Indica and Japonica brown rice and their malts. LWT-Food Science and Technology,
43(5), 784-791.
[5] Ayernor, G. S., & Ocloo, F. C. K. (2007). Physico-chemical changes and diastatic activity associated with

U
germinating paddy rice (PSB. Rc 34). African Journal of Food Science, 1, 37-41.
N
[6] Aggarwal, P., & David Dollimore, D. (1998). A thermal analysis investigation of partially hydrolyzed starch.
Thermochimica Acta, 319, 17-25.
A
[7] Limpisuta, P., & Jindal, V.K. (2002). Comparison of rice flour pasting properties using Brabender viscoamylo
M

graph and rapid visco analyser for evaluating cooked rice texture. Starch, 54, 350–357.
[8] Olugbile, A. O., Obadina, A. O., Atanda, A. O., Omemu, O. B., & Olatope, S. O. A. (2015). Physicochemical
Changes and Diastatic Activity Associated with Germination of ‘Boromo,’ a Paddy Rice Variety from Western
D

Nigeria. Journal of Food Processing and Preservation, 39(2), 116-122.

TE

[9] Kumar, N., Bansal, A., Sarma, G. S., & Rawal, R. K. (2014). Chemometrics tools used in analytical chemistry: An
overview. Talanta, 123, 186-199.
[10] Mobili, P., Londero, A., De Antoni, G., Gomez-Zavaglia, A., Araujo-Andrade, C., Avila-Donoso, H., …&
EP

Frausto-Reyes, C. (2010). Multivariate analysis of Raman spectra applied to microbiology: Discrimination of

microorganisms at the species level. Revista Mexicana de Física, 56(5), 378-385.
CC

[11] Abdi, H., & Williams, L. J. (2010). Principal component analysis. Wiley Interdisciplinary Reviews:
Computational Statistics, 2(4), 433-459.
[12] Quadir, N., Wani, S. A., Bhat, B. A., Wani, T. A., & Quraazah, A. (2012). Germination behavior of some
A

Kashmiri paddy cultivars. Journal of Chemical, Biological and Physical Sciences, 2, 1820-1829.
[13] Kalita, D., Bhattacharya, S., & Srivastava, B. (2018). Predicting enzymatic starch hydrolysis mechanism during
paddy malting by vibrational spectroscopy and multivariate calibration analysis. Food Chemistry, 259, 89-98.
[14] Labuschagne, M., Phalafala, L., Osthoff, G., & Biljon, A. V. (2014). The influence of storage conditions on starch
and amylose content of South African quality protein maize and normal maize hybrids. Journal of Stored Products
Research, 56, 16–20.
[15] Wu, F., Chen, H., Yang, N., Wang, J., Duan, X., Jin, Z., & Xu, X. (2013). Effect of germination time on
physicochemical properties of brown rice flour and starch from different rice cultivars. Journal of Cereal Science,
58, 263-271.
[16] Ludikhuyze, L., Ooms, V., Weemaes, C., & Hendrickx, M. (1999). Kinetic study of the irreversible thermal and
pressure inactivation of myrosinase from broccoli (Brassica oleracea L. Cv. Italica). Journal of Agricultural and
Food Chemistry, 47(5), 1794–1800.
[17] Lawless, H.T., & Heymann, H. (1998). Data relationships and multivariate applications. In: Sensory evaluation
of food: principles and practices. New York: Chapman and Hall. p 585–601.
[18] Atichokudomchai, N., Jane, J.-L., & Hazlewood, G. (2006). Reaction pattern of a novel thermostable α-amylase,
Carbohydrate Polymers, 64, 582–588.
[19] Jang, E-H., Lee, S-J., Hong, J-Y., Chung, H-J., Lee, Y-T., Kang, B-S., & Lim, S-T. (2016). Correlation between
physicochemical properties of japonica and indica rice starches. LWT - Food Science and Technology, 66, 530-537.
[20] Park, I-M., Ibanez, A.M., Zhong, F., & Shoemaker, C.F. (2007). Gelatinization and pasting properties of waxy and
non-waxy rice starches. Starch, 59, 388–396.

T
[21] Wokadalaa, O.C., Rayb, S.S., & Emmambux, M.N. (2012). Occurrence of amylose-lipid complexes in teff and

IP
maize starch biphasic pastes. Carbohydrate Polymers, 90, 616– 622.

R
[22] Bail, L.P., Bizot, H., Ollivon, M., Keller, G., Bourgaux, C., Buleon, A. (1999). Monitoring the crystallization of
amylose-lipid complexes during maize starch melting by synchrotron X-Ray diffraction. Biopolymers, 50, 99–110.

SC
[23] Kalita, D., Sarma, B., & Srivastava, B. (2017). Influence of germination conditions on malting potential of low
and normal amylose paddy and changes in enzymatic activity and physico chemical properties. Food
Chemistry, 220, 67-75.

U
[24] Musa, A. S. N., Umar, I. M., & Ismai, M. (2011). Physicochemical properties of germinated brown rice (Oryza
N
sativa L.) starch. African Journal of Biotechnology, 10, 6281-6291.
A
[25] Woo, H.-D., We, G.N., Kang, T.-Y., Shon, K.H., Chung, H.-W., Yoon, M.-R., Lee, J.-S. & Ko, S. (2015).
Physicochemical and gelatinization properties of starches separated from various rice cultivars. Journal of Food
M

Science, 80(10), 2208-2216.

[26] Chinma, C.E., Adewuyi, O., & Abu, J.O. (2009). Effect of germination on the chemical, functional and pasting
properties of flour from brown and yellow varieties of tigernut (Cyperusesculentus). Food Research International,
D

42, 1004–1009.
TE

[27] Noda, T., Nishiba, Y., Sato, T., & Suda, I. (2003). Properties of Starches from Several Low-Amylose Rice
Cultivars. Cereal Chemistry, 80, 193–197.
EP

[28] Eliasson, A.-C &Ljunger, G. (1988). Interactions between amylopectin and lipid additives during retrogradation in
a model system. Journal of Food and Agriculture, 44, 353–361.
[29] Nelles, E. M., Dewar, J., Bason, M. L., & Taylor, J. R. N. (2000). Maize starch biphasic pasting curves. Journal of
CC

Cereal Science, 31(3), 287-294.

[30] Morrison, W. R., & Azudin, M. N. (1987). Variation in the amylose and lipid contents and some physical
properties of rice starches. Journal of Cereal Science, 5(1), 35–44.
A

[31] Damodaran, S., Parkin, K. L., and Fennema, O. R. (2007). Food chemistry (4th ed.). United States: CRC press
[32] Ashogbon, A.O., & Akintayo, E.T. (2012). Morphological, functional and pasting properties of starches separated
from rice cultivars grown in Nigeria. International Food Research Journal, 19, 665-671.
[33] Falade, K.O., & Christopher, A. S. (2015). Physical, functional, pasting and thermal properties of flours and
starches of six Nigerian rice cultivars. Food Hydrocolloids, 44, 478-490.
[34] Kong, X., Zhu, P., Sui, Z., & Bao, J. (2015). Physicochemical properties of starches from diverse rice cultivars
varying in apparent amylose content and gelatinization temperature combinations. Food Chemistry, 172, 433–440.
[35] Mir, S. A., & Bosco, S. J. D. (2014). Cultivar difference in physicochemical properties of starches and flours from
temperate rice of Indian Himalayas. Food Chemistry, 157, 448-456.
[36] Patindol, J., Gu, X., & Wang, Y-J. (2009). Chemometric analysis of the gelatinization and pasting properties of
long-grain rice starches in relation to fine structure. Starch, 61, 3–11.
[37] Ye, L., Wang, C., Wang, S., Zhou, S., & Liu, X. (2016). Thermal and rheological properties of brown flour from
Indica rice. Journal of Cereal Science, 70, 270-274.

T
R IP
SC
U
N
RLA30 RLA35
A
M
D
TE
EP
CC
A

RNA30 RNA35

Fig. 1: DSC plots of RLA and RNA malted rice at different germination conditions
 3.5 120 1.4 120
Native 24 h
LR NR Temperature (°C) LR30 LR35 NR30 NR35
110 110
3.0 1.2

100 100
2.5 1.0
90 90

Temperature ( C)

Temperature ( C)
Viscosity (Pa-s)
Viscosity (Pa-s)

2.0 80 0.8 80

1.5 70 0.6 70

60 60
1.0 0.4

T
50 50

0.5 0.2
40 40

IP
0.0 30 0.0 30
0 3 6 9 12 15 0 3 6 9 12 15
Time (min) Time (min)

R
(a) (b)

SC
0.18 48 h 120 0.25 72 h 120
LR30 LR35 NR30 NR35
LR30 LR35 NR30 NR35
0.16 110 110

0.14 100
U
0.20
100
N
0.12 90 90
Temperature ( C)

Temperature ( C)
Viscosity (Pa-s)

Viscosity (Pa-s)

0.15
0.10 80 80
A
0.08 70 70
0.10
M

0.06 60 60

0.04 50 50
0.05
D

0.02 40 40

0.00 30 0.00 30
TE

0 3 6 9 12 15 0 3 6 9 12 15
Time (min) Time (min)

(c) (d)
EP

0.09 96 h 120 0.09 120 h 120

LR30 LR35 NR30 NR35 LR30 LR35 NR30 NR35
0.08 110 0.08 110
CC

0.07 100 0.07 100

0.06 90 0.06 90
Temperature ( C)

Temperature ( C)
Viscosity (Pa-s)

Viscosity (Pa-s)

0.05 80 0.05 80
A

0.04 70 0.04 70

0.03 60 0.03 60

0.02 50 0.02 50

0.01 40 0.01 40

0.00 30 0.00 30
0 3 6 9 12 15 0 3 6 9 12 15
Time (min) Time (min)

(e) (f)
Fig. 2: Pasting properties of RLA and RNA malted rice at different germination conditions: (a) Native rice,
(b) 24 h germination, (c) 48 h germination, (d) 72 h germination, (e) 96 h germination, and (f) 120
h of germination

T
R IP
SC
U
N
A
M
D
TE
EP
CC
A
 T
R IP
SC
U
N
A
M
D
TE
EP
CC
A

Fig. 3: PCA biplots of pasting and thermal properties of maltedRLA variety germinated at 30°C (top) and
35°C (bottom)
 T
R IP
SC
U
N
A
M
ED
E PT
CC
A

Fig. 4: PCA biplots of pasting and thermal properties of RNA malted paddy
germinated at 30°C (top) and 35°C
(bottom)

20
 Table 1. Changes in amylose and amylopectin contents in low amylose rice (RLA) and
normal
amylose rice (RNA) during germination
Time Amylose Amylopectin Starch Reducing sugar
(h)
30°C 35°C 30°C 35°C 30°C 35°C 30°C 35°C
Low amylose rice (RLA)
0 12.32±0.35 12.32±0.35 62.12±0.93 62.12±0.93 74.58±0.47 74.58±1.67 0.33±0.03 0.33±0.03
24 12.23±0.21 12.20±0.25 53.23±1.01 52.34±0.80 65.47±0.03 64.54±0.50 0.40±0.03 0.71±0.10

T
48 13.03±0.67 13.79±0.21 28.64±0.39 23.51±1.21 41.68±0.40 37.31±0.17 0.64±0.08 1.58±0.03

IP
72 14.61±0.28 14.73±0.42 23.78±0.78 17.82±1.13 38.39±1.13 32.55±0.32 2.03±0.08 2.22±0.03
96 15.46±0.21 17.69±0.33 18.40±0.73 13.10±0.85 33.86±0.73 30.79±0.17 3.24±0.20 4.09±0.05

R
120 16.53±0.21 21.10±0.28 14.15±0.12 6.86±0.61 30.69±0.02 27.96±0.17 4.80±0.25 7.02±0.13

SC
Normal amylose rice (RNA)
0 20.21±0.70 20.20±0.70 61.32±0.74 61.32±0.74 81.51±0.70 81.51±0.73 0.18±0.03 0.17±0.03
24
48
19.67±0.22
20.22±1.41
20.01±0.70
19.98±0.49
60.28±1.06
55.21±1.29
54.33±0.31
36.29±0.76 U
79.96±1.47
75.44±0.37
74.35±1.32
56.23±2.28
0.42±0.13
0.9±0.03
0.45±0.08
0.71±0.10
N
72 16.41±0.35 18.27±0.42 38.95±1.04 16.09±0.71 55.36±0.66 34.37±0.44 1.53±0.20 1.24±0.05
A
96 15.68±0.14 17.32±0.70 27.30±0.73 12.27±0.50 42.99±1.03 29.59±1.62 2.74±0.15 2.79±0.03
M

120 13.73±0.49 14.64±0.37 14.58±0.65 5.81±0.29 28.32±0.15 20.45±0.29 5.3±0.45 6.01±0.30

ED
E PT
CC
A

21
Table 2: Thermal properties of malted RLA and RNA varieties at different germination
conditions
Germination time Onset temperature Peak temperature Endset temperature
(h) (TO) (TP) (TC)
RLA30
0 64.08±0.34a 83.20±1.45a 100.10±0.16a
24 98.73±1.26f 117.4±1.59f 129.87±2.45f
48 92.37±0.56e 115.49±2.78e 128.23±0.98e
72 90.37±2.68d 114.39±0.38d 123.68±1.67c
96 87.27±0.89b 111.67±2.98c 124.95±0.23d

T
120 82.18±0.98b 99.46±0.29b 112.02±1.78b

IP
RLA35
0 64.08±0.81a 83.20±0.18a 100.10±0.93a

R
24 95.81±1.25f 111.49±2.76f 118.04±1.89e
48 88.90±0.87e 104.49±1.75e 113.12±0.27d

SC
72 85.27±2.13d 104.03±0.34d 112.39±0.56d
96 83.80±0.15c 102.38±1.36c 109.65±0.99c
120 82.35±2.67b 98.95±0.19b 108.02±1.78b

RNA30 U
N
0 65.73±1.34a 86.09±0.29a 100.13±1.14a
24 98.56±0.97f 108.98±1.48e 118.52±2.38e
A
48 92.54±1.56e 108.49±0.23e 120.03±0.45f
72 83.083±0.38d 103.52±1.29c 112.94±1.78d
M

96 82.18±2.67c 100.36±0.18b 111.12±0.15c

120 79.26±1.53b 97.49±0.33b 107.65±2.13b

RNA35
ED

0 65.73±0.98a 86.09±0.98b 100.13±0.76b

24 83.54±2.59d 109.19±1.45f 127.54±1.34e
48 78.99±0.31c 103.54±2.98e 112.90±0.34d
PT

72 71.73±1.76b 96.63±0.17d 105.37±1.78c

96 71.00±2.67b 88.81±0.45c 99.00±2.89b
120 66.06±0.56a 82.27±1.67a 91.73±1.41a
E

Values in the same column with different subscripts are significantly different at p<0.05
CC

Abbreviations
TO: Onset temperature; TP: Peak temperature; TC: Conclusion temperature.
A

RLA30 and RLA35 indicate that germination of RLA variety paddy was done at temperature of 30 and 35°C,
respectively.

RNA30 and RNA35 indicate that germination of RNA variety paddy was done at temperature of 30 and 35°C,
respectively

22
 Table 3: Pasting properties of malted RLA and RNA rice at different germination
conditions
Time (h) PT PV1 PV2 HPV BDV SBV CPV
RLA30 °C mPas mPas mPas mPas mPas mPas
a
0 71±1 c 0±0 1757±3 e
1092±1 f
665±5 d
686±2b
1785±1e
24 69±2b 226±1f 255±1d 200±2e 55±0c 175±2ab 374±4d
48 68±0b 92±0e 100±1c 83±0d 17±1b 78±0ab 160±1c
72 65±0a 33±1d 40±0b 30±1c 10±1a 19±0a 51±0b
96 65±0a 28±1c 30±1a 21±0b 9±1a 13±1a 36±1a
120 65±0a 25±1b 28±1a 20±1c 8±2a 12±1a 32±2a

T
RLA35

IP
0 71±1a 0±0a 1757±3e 1092±1e 665±5c 686±2b 1785±1e
24 70±7a 305±7d 320±6d 258±12d 62±4b 212±8c 471±1d
48 70±7a 67±0c 82±0c 68±0c 15±0a 53±1b 121±2c

R
72 66±6a 32±2b 44±0b 38±0b 6±0a 20±2a 57±2b
96 66±3a 25±0b 32±0a 24±1a 7±1a 11±0a 35±0a

SC
120 66±2a 22±1b 27±1a 22±1a 5±0a 10±1a 32±0a

Time (h) PT PV HPV BDV SBV CPV

RNA30
0
°C
75±0b
mPa-s
3041±2f
mPa-s
806±3f U
mPa-s
2235±5e
mPa-s
2113±2e
mPa-s
2919±4e
N
24 72±1b 987±1e 407±4e 583±2d 821±1d 1228±3d
48 71±1b 157±3b 47±1c 111±3c 79±2c 126±6c
A
72 70±0a 146±1d 45±0d 142±1b 64±0b 118±1b
96 70±0a 79±1b 41±1b 41±0a 24±1a 61±0a
68±1a 73±1a 31.5±2a 38.5±1a 20±3a 55±1a
M

120
RNA35
0 75±0b 3041±2e 806±3e 2235±5f 2113±2f 2919±4e
24 72±1b 624±6d 241±6d 383±0e 442±1e 683±4d
ED

48 72±1a 151±4c 45±1c 106±2d 35±0d 76±4c

72 70±0a 132±3b 45±1c 87±2c 32±1c 81±2c
96 70±0a 79±0a 34±0b 45±0b 24±1b 58±1b
120 68±1a 75±0a 32±0a 43±0a 15±0a 42±0a
PT

Values in the same column with different subscripts are significantly different at p<0.05

Abbreviations
E

PT: Pasting temperature; PV1: First peak viscosity; PV2: Second peak viscosity; HVP: Hot paste viscosity;
BDV: Breakdown viscosity; SBV: Set back viscosity; CPV: Cold paste viscosity.
CC

RLA30 and RLA35 indicate that germination of RLA variety paddy was done at temperature of 30 and 35°C,
respectively.
RNA30 and RNA35 indicate that germination of RNA variety paddy was done at temperature of 30 and 35°C,
A

respectively

23
 Table 4: Parameters related to pasting and thermal properties as per second-order
kinetic model
Sample RVA A0 k (h-1) R2 RMSE

T
parameter
RLA30 PT 71 1.31x10-05 0.865 2.06x10-04

IP
PV2 1757 3.05x10-04 0.946 3.56x10-03
HPV 1785 2.52x10-04 0.925 3.65x10-03

R
CPV 1092 4.21x10-04 0.930 5.74x10-03

1.02x10-05 2.14x10-04

SC
RLA35 PT 71 0.829
PV2 1757 3.01x10-04 0.970 2.58x10-03
HPV 1785 2.53x10-04 0.937 3.33x10-03
CPV 1092 3.73x10-04 0.955 3.99x10-03

RNA30 PT 75 1.17x10-05 0.883

U 1.59x10-04
N
PV 3041 1.13x10-04 0.941 1.37x10-03
HPV 2919 1.47x10-04 0.930 2.00x10-03
A
CPV 806 2.65x10-04 0.884 4.26x10-03

RNA35 PT 75 7.00x10-06 0.721 1.54x10-04

M

PV 3041 1.14x10-04 0.958 1.11x10-03

HPV 2919 1.88x10-04 0.928 2.44x10-03
CPV 806 2.80x10-04 0.894 4.15x10-03
ED

Thermal a b c d R2 RMSE
properties
RLA30 TO 64.080 6.45E+05 6267 12.62 0.994 1.462
PT

TP 83.250 7.75E+05 4968 20.56 0.968 3.780

TC 100.100 1.07E+06 6878 19.65 0.946 4.230

RLA35 TO 64.080 8.07E+05 8230 14.55 0.982 2.271

E

TP 83.200 8.38E+05 7416 8.83 0.979 2.163

TC 100.100 2.20E+06 8400 17.26 0.982 1.276
CC

RNA30 TO 65.730 6.78E+05 6486 18.71 0.975 2.819

TP 86.090 1.71E+05 1406 2.85 0.996 0.899
TC 100.100 1.06E+06 7883 1.661 0.971 1.959
A

RNA35 TO 65.730 6.63E+05 7431 21.71 0.975 1.768

TP 86.090 1.05E+05 7755 4.22 1.000 0.375
TC 100.100 8.66E+05 6210 27.26 0.990 2.049
Abbreviations

24
PT: Pasting temperature; PV1: First peak viscosity; PV2: Second peak viscosity; HVP: Hot paste viscosity;
BDV: Breakdown viscosity; SBV: Set back viscosity; CPV: Cold paste viscosity. TO: Onset temperature; TP:
Peak temperature; TC: Conclusion temperature.

RLA30 and RLA35 indicate that germination of RLA variety paddy was done at temperature of 30 and 35°C,
respectively.

RNA30 and RNA35 indicate that germination of RNA variety paddy was done at temperature of 30 and 35°C,
respectively

a, b, c and d are the model parameters of Eq (1).

R2 is coefficient of determination

T
RMSE is root mean square error

R IP
SC
U
N
A
M
ED
E PT
CC
A

25
 Table 5: Lower-half correlation matrix among pasting and thermal properties
PT PV1 PV2 HPV BDV SBV CPV TO TP TC Amys Amylp
RLA30
PT 1**
PV1 0.315
PV2 0.806 -0.277
HPV 0.827* -0.242 0.999**
BDV 0.771 -0.332 0.998** 0.995**
SBV 0.868* -0.166 0.993** 0.997** 0.985**
CPV 0.842* -0.215 0.998** 1** 0.992** 0.999**
TO 0.197 0.949** -0.413 -0.380 -0.465 -0.307 -0.354

T
TP -0.442 0.603 -0.835* -0.819* -0.859* -0.784 -0.808 0.753
TC -0.359 0.650 -0.789 -0.771 -0.816* -0.730 -0.757 0.795 0.991

IP
Amys -0.895* -0.511 -0.561 -0.588 -0.516 -0.642 -0.607* -0.489 0.050 -0.024 1**
Amylp 0.966** 0.346 0.797 0.818* 0.761 0.858* 0.833** 0.193 -0.396 -0.329* -0.889* 1**

R
RLA35
PT
0.710

SC
PV1
PV2 -0.023 -0.143
HPV 0 -0.012 1**
BDV -0.059 -0.182 0.999** 0.998**
SBV 0.074 -0.020 0.992** 0.995** 0.987**
CPV
TO
0.029
0.532
-0.080
0.690
0.998**
-0.799
0.999**
-0.783
0.995**
-0.823*
0.998**
-0.720 -0.760U
N
TP 0.483 0.674 -0.811 -0.796 -0.834* -0.735 -0.773 0.995**
TC 0.608 0.745 -0.716 -0.698 -0.744 -0.630 -0.672 0.984 0.982**
A
Amys -0.686 -0.421 -0.552 -0.569 -0.525 -0.614 -0.587 0.058 0.066 -0.100
Amylp 0.451 0.419 0.825* 0.839* 0.801 0.886* 0.858 -0.321 -0.339 -0.198 -0.836 1**
M

PT PV HPV BDV SBV CPV TO TP TC Amys Amylp

RNA30
PT 1**
0.877*
ED

PV
HPV 0.904* 0.983**
BDV 0.861* 0.998** 0.970**
SBV 0.894* 0.997** 0.995** 0.989**
CPV 0.861* 0.998** 0.970** 1** 0.989**
PT

To 0.155 -0.282 -0.152 -0.325 -0.228 -0.325

Tp -0.318 -0.708 -0.604 -0.739 -0.665 -0.739 0.873*
Tc -0.205 -0.638 -0.539 -0.667 -0.596 -0.667 0.898 0.987**
Amys 0.896* 0.587 0.627 0.568 0.608 0.568 0.496* 0.080 0.211
E

Amylp 0.905* 0.628 0.680 0.605 0.653 0.605 0.519* 0.069 0.180 0.982** 1**
CC

RNA35
PT 1**
PV 0.636
HPV 0.701 0.996**
A

BDV 0.611 0.999** 0.992**

SBV 0.657 1** 0.998** 0.998**
CPV 0.669 0.999** 0.999** 0.997** 1**
To 0.167 -0.355 -0.289 -0.378 -0.346 -0.331
Tp 0.195 -0.249 -0.188 -0.270 -0.244 -0.229 0.969*
Tc 0.439 -0.075 -0.001 -0.101 -0.064 -0.047 0.944 0.964*
Amys 0.430 0.492 0.523 0.480 0.489 0.498 0.539 0.658 0.700 1**

26
Amylp 0.742 0.749 0.790 0.733 0.754 0.764 0.349 0.430 0.586 0.857* 1**
* Significant at p=0.05 ** Significant at p=0.01 All other are non-significant at p=0.05
Abbreviations
PT: Pasting temperature; PV1: First peak viscosity; PV2: Second peak viscosity; HVP: Hot paste viscosity; BDV:
Breakdown viscosity; SBV: Set back viscosity; CPV: Cold paste viscosity. TO: Onset temperature; TP: Peak temperature;
TC: Conclusion temperature; Amys: Amylose; Amylp: Amylopectin.
RLA30 and RLA35 indicate that germination of RLA variety paddy was done at temperature of 30 and 35°C, respectively.
RNA30 and RNA35 indicate that germination of RNA variety paddy was done at temperature of 30 and 35°C, respectively

T
R IP
SC
U
N
A
M
ED
E PT
CC
A

27
 Appendix
Appendix Table 1: ANOVA table for pasting properties
Source Dependent Variable F-value p-value
Corrected Model PT 6.49 .000
PV 278993.02 .000
HPV 26518.62 .000
BDV 45124.84 .000
SBV 122.57 .000
CPV 325414.28 .000
Intercept PT 65735.20 .000
PV 2406588.71 .000
HPV 281966.95 .000

T
BDV 276967.69 .000
SBV 656.54 .000

IP
CPV 2988404.09 .000
Variety (LR and NR) PT 72.09 .000
PV 238591.03 .000
HPV 1262.26 .000

R
BDV 97420.07 .000
SBV 331.88 .000
CPV 197927.43 .000

SC
Temperature (30 and 35°C) PT 0.21 .653
PV 1731.79 .000
HPV 168.83 .000
BDV 233.75 .000
SBV 8.56 .007

Germination time (h)

CPV
PT
PV U 5805.61
9.95
1149601.12
.000
.000
.000
N
HPV 117201.49 .000
BDV 145956.50 .000
SBV 293.27 .000
A
CPV 1356736.84 .000
Variety * Temp PT 9.20 .006
PV 3561.40 .000
M

HPV 536.30 .000

BDV 343.75 .000
SBV 0.32 .579
CPV 8520.00 .000
Variety * Time PT 1.20 .341
ED

PV 81214.90 .000
HPV 3838.74 .000
BDV 41625.30 .000
SBV 193.15 .000
CPV 87944.86 .000
PT

Temp * Time PT 0.40 .844

PV 966.22 .000
HPV 97.44 .000
BDV 135.07 .000
SBV 2.65 .048
E

CPV 3097.39 .000

Variety * Temp * Time PT 2.00 .116
PV 2808.80 .000
CC

HPV 454.49 .000

BDV 257.88 .000
SBV 6.60 .001
CPV 6676.00 .000
Error PT
A

PV
HPV
BDV
SBV
CPV
Total PT
PV
HPV
BDV

28
 SBV
CPV
Corrected Total PT
PV
HPV
BDV
SBV
CPV
Note: Variety corresponds to amylose-amylopectin content

T
R IP
SC
U
N
A
M
ED
EPT
CC
A

29
 Appendix Table 2: ANOVA table for thermal properties
Source Dependent F-value p-value
Variable
Corrected Model TO 4677.96 .000
TP 4444.02 .000
TC 1885.27 .000
Intercept TO 6188099.61 .000
TP 9207766.91 .000
TC 5175606.59 .000
Temperature (30 and 35°C) TO 10143.46 .000
TP 9332.78 .000

T
TC 5969.33 .000
Germination time (h) TO 15384.86 .000

IP
TP 14643.75 .000
TC 5169.50 .000
Variety (LR and NR) TO 9403.72 .000

R
TP 8184.68 .000
TC 3812.01 .000
Temp * Time TO 442.57 .000

SC
TP 620.52 .000
TC 550.34 .000
Temp * Variety TO 3908.59 .000
TP 3.20 .086

Time * Variety
TC
TO U 394.04
759.35
.000
.000
N
TP 1071.53 .000
TC 449.48 .000
A
Temp * Time * Variety TO 240.67 .000
TP 602.56 .000
TC 467.83 .000
M

Error TO
TP
TC
Total TO
ED

TP
TC
Corrected Total TO
TP
PT

TC
Note: Variety corresponds to amylose-amylopectin content
E
CC
A

30
 Table Appendix 3: Zero and first order kinetic parameters related to pasting properties
Paddy RVA A0 Zero order First order
parameter
k0 R2 RMSE k1 R2 RMSE
RLA30 PT 71 -0.061 0.859 0.963 -8.90x10-04 0.862 0.014

PV 1757 -19.330 0.115 645.000 -4.24 x10-02 0.772 0.769

CPV 1785 -19.410 0.226 605.400 -4.00 x10-02 0.866 0.578

T
HPV 1092 -11.920 0.179 383.000 -4.05x10-02 0.827 0.653

IP
RLA35 PT 71 -0.048 0.832 0.984 -6.96x10-04 0.831 0.014

R
PV 1757 -19.300 0.159 628.200 -4.23x10-02 0.791 0.744

SC
CPV 1785 -19.390 0.283 584.800 -4.01x10-02 0.871 0.577

HPV 1092 -11.870 0.238 368.300 -4.00x10-02 0.833 0.629

RNA30 PT 75 -0.061 0.862 U

0.882 -8.40x10-04 0.873 0.012
N
PV 3041 -32.720 0.340 956.700 -3.74x10-02 0.815 0.650
A
CPV 2919 -31.390 0.451 856.200 -3.95x10-02 0.835 0.677
M

HPV 806 -8.434 0.514 221.900 -3.27x10-02 0.751 0.693

ED

RNA35 PT 75 -0.038 0.716 0.832 -5.13x10-04 0.718 0.011

PV 3041 -33.030 0.178 1064.000 -3.80x10-02 0.740 0.740

CPV 2919 -32.020 0.215 1012.000 -4.28x10-02 0.734 0.873

PT

HPV 806 -8.581 0.282 260.500 -3.36x10-02 0.698 0.727

E
CC
A

31