Food Chemistry Advances 2 (2023) 100190

Contents lists available at ScienceDirect

Food Chemistry Advances

journal homepage: www.elsevier.com/locate/focha

Thermal treatment in combination with laminated packaging under

modified atmosphere enhances the shelf life of pearl millet flour
P.G. Padmaja∗, A. Kalaisekar, R. Venkateswarlu, S. Shwetha, B. Dayakar Rao, V.A. Tonapi
ICAR-Indian Institute of Millets Research, Rajendranagar, Hyderabad, Telangana 500030, India

a r t i c l e i n f o a b s t r a c t

Keywords: Pearl millet grain is storable for months without development of rancidity, while milled flour has very short shelf
Pearl millet life of 5 to 7 days due to high fat content and lipolytic action of enzymes. Apart from enzymatic degradation
Packaging of lipids and c-glycosyl flavones, oxygen present in air promotes oxidation of free fatty acids (FFA) leading to
Rancidity
production of peroxides which impart off flavours and bitterness. To control both enzymatic and oxidative effects,
Shelf life
implementation of an integrated approach of thermal treatment, efficient packaging, and modified atmosphere
Thermal
was used for shelf life extension. Heat treatment of grains at 150 °C for 30 min before milling and storage of
flour in laminated pouches under nitrogen atmosphere was found to extend shelf life by 60 days compared to
untreated samples by decreasing FFA value by 10–15 fold and peroxide value by 2 fold. Genotype, thermal
treatment, packaging material and atmosphere were found to be significantly different for rancidity parameters.

1. Introduction oxidation in the presence of oxygen and moisture resulting in undesir-

Pearl millet (Pennisetum glaucum (L.) R. Br.) is a grass cultivated in
tropical countries. Grain is used as food for human consumption, green
fodder and dry stover as feed for livestock in India, Africa and South
Asia. It occupies 30 million ha worldwide (Raheem et al., 2021). More
than 90 million people are dependent for nutritional and livelihood se-
curity in African and Asian countries (Serba et al., 2020). In India, after
rice and wheat, it is the most important food crop in area and produc-
tion. It is cultivated in an area of 6.93 million ha with an average pro-
duction of 8.61 million tones and productivity of 1243 kg/ha in India
(Directorate of Millets Development, 2020).
Pearl millet grain contains 13.6% crude protein, 5–7% fat and 63.2%
starch (Goswami et al., 2020). It is rich in iron and zinc compared to
sorghum, wheat, rice and maize (Longvah et al., 2017). Pearl millet is
used to prepare traditional fermented porridges. Malted grain is used for
brewing traditional wines and beers in Africa. In Cameroon, gruels and
steamed cakes are used for feeding infants and children. Malted pearl
millet in combination with legumes is used for preparing weaning foods
(Gomez & Gupta, 2003).
Despite its nutrient profile, limited use of flour is due to its poor
keeping quality. Pearl millet flour is rich in fat and comprises of 74%
unsaturated fatty acids (oleic, linoleic and linolenic) in its fatty acid pro-
file (Nantanga, 2006). Lipolytic enzymes are concentrated in the aleu-
rone layer, pericarp and germ. The lipases are very active leading to
release of free fatty acids. The released unsaturated fatty acids undergo
able flavuor. Lipase activity is relatively high in pearl millet compared
to most of the cereal grains (Galliard, 1999). Apart from lipases, per-
oxidases (Reddy et al., 1986) and polyphenol oxidase (PPO) are also
involved in deterioration of flour quality. Milling of pearl millet grains
into flour brings enzymes into contact with their substrates. Under high
moisture and oxygen, the products of enzymatic reaction give rise to off
odours and bitter taste. Oxidative rancidity results in hydroperoxides
which subsequently breakdown to produce volatile compounds with
rancid odor. Apart from lipolytic enzymes, peroxidases release pheno-
lic aglycones from c-glycosyl flavones producing bitter taste (Eskin &
Przybylski, 2001). Bitter tasting compounds are produced by polyphe-
nol oxidase (PPO) through enzymatic browning reaction. This short shelf
life of flour is the major hurdle for its utilization in food industry and
domestic consumption. The shelf life of pearl millet flour was improved
by pearling (Malviya, 2015), fermentation (Chinenye et al., 2017), de-
fatting (Kapoor & Kapoor, 1990), use of antioxidants (Abdalgader et al.,
2019), and heat treatments to control oxidation of fat. Dry heat treat-
ment in hot air oven at 100 °C for 10 min to 2 h (Arora et al., 2002;
Kadlag et al., 1995; Patel & Parameswaran, 1992) was reported to ex-
tend shelf life of flour by 21- 28 days. Similarly, blanching of pearl millet
grains at 98 °C for 10 to 20 s (Kadlag et al., 1995) or microwave treat-
ment of grains at 18–20% moisture for 80–90 s reaching a final tem-
perature of 107.6–110 °C (Mohajerkhorasani et al., 2019; Yadav et al.,
2012a) could extend shelf life of pearl millet flour by 30 days. These
treatments were applied to control enzymatic degradation while ox-

∗
Corresponding author.
E-mail address: padmaja@millets.res.in (P.G. Padmaja).

https://doi.org/10.1016/j.focha.2023.100190
Received 29 May 2022; Received in revised form 2 January 2023; Accepted 12 January 2023
2772-753X/© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/)
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
idative rancidity is also an important contributing factor for spoilage
of pearl millet flour. Oxidative rancidity can be controlled by remov-
ing oxygen surrounding the product and refilling with inert gas like
nitrogen. This method is referred as modified atmosphere packaging
(Hintlian & Hotchkiss, 1986). Nitrogen (N2 ) apart from being inert, has
low solubility in water and prevents the growth of aerobic microorgan-
isms (Mullan & McDowell, 2011). The stability of modified atmosphere
inside the packaging material is dependent on its gas transmission rate.
Storage of flour in sealed HDPE package was found to prevent gener-
ation of free radicals and proliferation of microbes thereby prolonging
the shelf life (Ogori et al., 2013; Varsha & Narayanan, 2017). The efforts
to study the development of rancidity in pearl millet flour were focused
on individual techniques by earlier researchers. In the present study, we
applied a simple and integrated approach to control both enzymatic and
oxidative rancidity for extending shelf life of pearl millet flour. A com-
bination of thermal treatment of pearl millet grains and packaging the
flour in metalized laminated pouches under nitrogen atmosphere has
extended the shelf life by two months.
2. Materials and methods
2.1. Pearl millet grains
Four kilograms of pearl millet whole grains of three genotypes HHB
67, MPMH 21 and Dhanshakthi were obtained from Seed Science depart-
ment of ICAR - Indian Institute of Millets Research, Hyderabad, India.
These grain samples were harvested during September, 2019 and were
stored in cloth bags.
2.2. Treatments
2.2.1. Refrigeration (Refn)
Shelf life of millet flour was determined at different storage intervals
(7, 15, 30, 45 and 60 days) under room temperature (27 °C) and at
refrigeration (4 °C).
2.2.2. Thermal treatment (Th)
The grain sample was divided into two portions of about 2 kg each.
One portion was spread on glass petriplates and placed in a hot air
oven set to 150 °C and heated for 30 min. Another untreated por-
tion was used as control. This time and temperature combination was
found to be organoleptically acceptable from various time and tem-
perature combinations tested. To standardize the temperature, initially
grains were treated for 10, 20 and 30 min at 175 °C. Lipase activity
was found to decrease significantly at 30 min (0.90 U/g) compared
to 10 (7.32 U/g) & 20 min (1.92 U/g). However, the grains were not
organoleptically acceptable. Hence, temperatures of 50, 100, 150 were
tested for 30 min and found that lipase activity was least at 150 °C and
grains were organoleptically acceptable (unpublished data). Based on
this trial, the combination of 150 °C for 30 min was chosen for further
experiments. Each treatment was replicated thrice. The samples were
ground to flour using a cyclone sample mill (Retsch, Germany) fitted
with 0.1 mm screen. Appropriate amounts of flour from the two treat-
ments were divided into fresh and stored flours. Stored flour was sealed
in different packaging materials and stored under ambient conditions
for two months of storage.
2.2.3. Packaging materials
Flour of pearl millet samples was stored in different packaging
materials viz., LDPE (low density polyethylene), HDPE (high density
polyethylene) and metalized laminated pouches (Lmn) along with con-
trol (zip-lock polyethylene pouches) in three replications. The nitro-
gen gas permeability of laminated and HDPE pouches was 3.85 and
60.72 cm3 /m2 .24 h.0.1 MPa respectively. However, gas permeability
determination for LDPE packaging failed due to very high permeability.
Keeping quality of the flour was analysed at different storage intervals
(7, 15, 30, 45 and 60 days).
2
Food Chemistry Advances 2 (2023) 100190
2.2.4. Modified atmosphere packaging (MAP)
Nitrogen (N2 ) gas was applied to packaged flour to alter the atmo-
sphere surrounding the flour and stored in different packaging mate-
rials viz., LDPE, HDPE and Laminated pouches along with control in
three replications. Keeping quality of the flour was analysed at different
storage intervals.
2.2.5. Combination treatments
Thermal treatment, packaging and modified atmosphere individu-
ally and in combinations was used to extend the shelf life of pearl mil-
let flour. The combination treatments are HDPE with Thermal treat-
ment (HDPE_Th), HDPE with N2 (HDPE _N), HDPE with Thermal treat-
ment and N2 (HDPE_Th_N), LDPE with N2 (LDPE_N), LDPE with Th
(LDPE_Th), LDPE with Th and N2 (LDPE_Th_N), Lamination with N2
(Lmn_N), Lamination with Th (Lmn_Th), Lamination with Th and N2
(Lmn_Th_N).
The stored flour of pearl millet genotypes was analyzed at different
storage intervals viz., 7, 15, 30, 45 and 60 days. Keeping quality of
millets flour was evaluated using alcoholic acidity, free fatty acid value,
peroxide value and lipase activity.
2.3. Alcoholic acidity
Alcoholic acidity (AA) was determined immediately after sample ex-
traction using standard method (IS 12711, 1989). Around 5 g of flour
sample was incubated with 50 mL of neutral ethyl alcohol for 24 h with
occasional stirring. Alcoholic extract was filtered through Whatmann
No.1 filter paper. A portion of the extract (10 mL) was titrated with
standardized sodium hydroxide solution (0.05 molL−1 ) using phenolph-
thalein as indicator. A blank was run in parallel and subtracted from the
samples. Alcoholic acidity was expressed as grams of H2 SO4 per 100 g
of dry sample.
2.4. Peroxide value
Sample extraction was performed using modified method of
Hare and Radin (1978). Around 4 g of pearl millet flour was mixed
with 20 mL of hexane: isopropanol (3:2 v/v) in a glass tube and rotated
for 10 min at room temperature (27 °C). The mixture was allowed to
settle at 4 °C in dark and the supernatant was decanted into a 50 mL
glass centrifuge tube. To this 10 mL of 6.7% (w/v) sodium sulfate was
added and inverted several times. This was subjected to centrifugation
at 1238X g for 10 min at 4 °C. The supernatant was transferred into a
glass test tube and the solvent was evaporated under nitrogen (Rapid
Mini, Crescent Scientific, India) to get oil residue.
Peroxide value (PV) was determined as per AOCS Cd 8b; 90, 2011.
The extracted oil was dissolved in 30 mL of acetic acid: isooctane (93:2)
in a flask. While swirling the flask, 0.5 mL of saturated solution of potas-
sium iodide was added and swirling continued for one minute. The re-
leased iodine was titrated using 0.01 molL−1 standardized sodium thio-
sulphate solution using starch as an indicator and titration was contin-
ued until the blue color disappeared indicating the end point. Peroxide
value was expressed as milliequivalents of peroxide/kg sample.
2.5. Free fatty acid value (FFA)
Free fatty acid value was quantified as described by Kwon and
Rhee (1986). Pearl millet flour of 1.5 g was extracted twice with 15 mL
of hexane. The pooled extracts were evaporated under nitrogen at 40 °C
(Rapid Mini, Crescent Scientific, India). The residue was dissolved in
5 mL of isooctane. To the mixture 1 mL of 5% (w/v) cupric acetate (pH
adjusted to 6.1 with pyridine) was added and shaken vigourously for
1 min. The tubes were centrifuged at 1000X g for 1 min. The absorbance
of sample was read at 750 nm and compared with the absorbance of oleic
acid standard solution prepared in isooctane (1–10 mmolL−1 ). FFA value
was expressed as μEq/1.5 g sample.
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
2.6. Lipase activity assay
The flour of three genotypes was defatted in petriplates with hexane
(1:10 w/v, three times) for 30 min on an orbital shaker at 140 rpm.
Residual hexane was removed by evaporation for 10 min at room tem-
perature. One gram of the defatted flour was taken into two test tubes.
Olive oil and distilled water were added to both tubes and mixed thor-
oughly. The lipids from the blank were extracted immediately. About
5 mL of hexane was added into the tube and vortexed and then cen-
trifuged at 1000X g for 3 min. The supernatent was decanted into a
30 mL glass test tube and the process was repeated two times. The hex-
ane extracts were pooled and evaporated under nitrogen at 40 °C (Rapid
Mini, Crescent Scientific, India). The residue was dissolved in 4.0 mL of
(iii) Effect of each genotype, each treatment and that of each storage
isooctane. The other test tube was incubated for 4 h at 40 °C. Lipids were
extracted as described for the blank. Free fatty acids were quantified as
per Kwon and Rhee (1986). The tubes were centrifuged at 1000X g for
1 min and the absorbance at 715 nm was taken in a spectrophotome-
ter. The absorbance of the sample was compared with the absorbance of
oleic acid standards prepared in isooctane (1–10 mmolL−1 ). Lipase ac-
tivity (LA) was expressed as units/gram (U/g), where 1.0 U was defined
as the micromoles of fatty acid liberated per hour (Eq. (1)).
[ ]
Lipase act ivit y = 1000 (4 + v)(Af − Ai)∕𝜖 tls (1)
Where 1000 = conversion factor from mol/L to μEq/mL
4 = volume of isooctane used to dissolve fat (mL)
v = volume of olive oil added (mL)
Af = absorbance of sample after incubation at 715 nm
Ai = absorbance of blank after incubation at 715 nm
€ = molar absorptivity of oleic acid at 715 nm (M−1 .cm−1 ) - value
t= incubation time (h)
l = path length (1 cm for a standard cuvette) and
s= sample weight (g)
2.6.1. Optimization of conditions
Lipase assay conditions were optimized using HHB 67 flour to yield
maximum lipase activity. The quantity of olive oil and water varied be-
tween 0.2 to 2.0 mL and 0.05 to 0.25 mL respectively. Incubation tem-
perature varied between 20 and 60 °C and incubation time ranged from
4 to 24 h. Lipase activity was maximum between 0.4 and 0.8 mL of oil
per gram of defatted sample (provided as supplementary data). Opti-
mal water concentration was found to be from 0.10 to 0.15 mL per g of
defatted sample. The optimal temperature for incubation was 40 °C.
2.7. Statistical analysis
2.7.1. Effects of treatment, storage period and genotype on rancidity
parameters in pearl millet flour
A sub-sub-plot experimental design was framed based on the proce-
dure explained by Altman and Krzywinski (2015) for laboratory con-
ditions. As per the requirements of the experimental design, we main-
tained two lots of pearl millet flour as blocking units, three genotypes of
pearl millet as whole plot (WP) factors, 15 treatments as sub plot (SP)
factors, and time-period intervals (0, 7, 15, 30, 45 and 60 days) as sub-
sub plot (SSP) factors. Therefore, in this experimental design, flour of
each genotype was subjected to 15 treatments, and in turn each treat-
ment was observed for four important rancidity parameters at six differ-
ent periods of storage. The details are given as supplementary material
in tabular form.
For the statistical analysis, we used R package ‘agricolae’ for sub-sub-
plot data to test the significance among all three levels of plot factors.
The significance levels were compared using interquartile ranges of the
data. Unlike means of observations, interquartile ranges of data sets are
not affected by outliers. Therefore, interquartile ranges of response vari-
ables were used for comparing plot factors. The results of the compar-
isons of interquartile ranges were presented as graphical representations
3
Food Chemistry Advances 2 (2023) 100190
in the results section. The results of sub-sub-plot design are useful in ex-
plaining the following three levels of comparisons of test factors.
The results of sub-sub-plot design are useful in explaining the fol-
lowing effects,
(i) ANOVA to test if the statistical significance in genotype, treatments,
and in storage time periods. Similarly, interaction effects between
genotype, treatment, and storage time periods on response variables
(rancidity parameters viz., FFA, AA, PV and LA) were also tested.
(ii) Then, ANOVA for two-factor (treatment and storage time period) ef-
fects within each genotypes were analysed to see if treatments and
storage time periods were significant within each of three pearl mil-
let genotypes.
time period on rancidity parameters are explained and presented in
graphical form. This explains how the effect of each component of
test factors varies.
2.7.2. Important contributing factors (rancidity parameters) to the
variability of treatments, storage periods, and genotypes
The role of rancidity parameters (response variables) viz., AA, FFA,
LA, and PV, in creating variability among WP, SP and SSP factors was
analysed with principal component analysis (PCA) biplot projections us-
ing R package ‘factoextra’. Most important treatments which could be
relied upon for improving shelflife of pearl millet flour during storage
were identified using supervised k-means clustering of treatments into
three clusters with linear projection of response variables. The treat-
ments were clustered using principal component 1 (PC1) and principal
component 2 (PC2).
PCA analyses results are presented in graphical form for the three test
factors along with linear projections of rancidity parameter variables.
The genotypes, treatments, and storage periods are grouped in a biplot
using first two principal component (PC) dimensions. The biplots thus
created explain the amount of variability in two PC dimensions. The
linear projections of variables show the role of rancidity parameters in
the biplot groupings of test factors.
2.7.3. Important treatments that are useful to increase the shelflife of pearl
millet flour
K-means clustering of treatments on a plot of PC1 and PC2 explains
the most useful treatments in improving storability of millet flour. The
graphical visualization of k-means clustering with linear projections was
produced using Python-Orange 3 (Demsar et al., 2013).
3. Results and discussion
Overall effects of genotype, treatments and storage periods on ran-
cidity parameters (alcoholic acidity, free fatty acid value, lipase activity
and peroxide value) are presented.
3.1. Effects of treatment, storage period and genotype on rancidity
parameters (FFA, AA, PV, and LA) in pearl millet flour
The intervention measures in terms of treatments, storage period and
genotypes to increase the shelf life of pearl millet flour were tested for
their effect on rancidity parameters. All the factors showed individual
as well as interaction effects significant with all response variables (ran-
cidity parameters).
3.1.1. Effect on free fatty acid value (FFA)
FFA measures the release of free fatty acids from triacylglycerols due
to action of lipolytic enzymes released during the process of milling. FFA
undergoes oxidation leading to formation of peroxides. Peroxides impart
off-flavours and bitterness to the flour.
Overall effect of treatment, storage period and genotype on FFA
showed significant variations between genotypes (df=2, p = 2e-16,
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
sig.0.001), between treatments (df=14, p = 2e-16, sig.0.001), and be-
tween storage periods (df=5, p = 2e-16, sig.0.001) (ANOVA table pro-
vided as supplementary data). The interaction effects between geno-
types and treatments (df=28, p = 2e-16, sig.0.001), between genotypes
and storage period (df=10, p = 2e-16, sig.0.001), between treatments
and storage period (df=70, p = 2e-16, sig.0.001), and between geno-
types, treatments and storage period (df=140, p = 2e-16, sig.0.001) were
all significant for FFA.
The influence of treatment and storage period on the rancidity pa-
rameter FFA in each genotype was analysed with two-way ANOVA
(ANOVA table provided as supplementary data). Results showed sig-
nificant effects individually as well as interactively on response vari-
ables in all the three genotypes. In the genotype Dhanshakthi, FFA
showed significant variations due to the effect factors viz., treat-
ment (F(14, 270) = 26,090, p = 0, 𝜂G2 = 0.999), and storage pe-
riod (F(5, 270) = 82,658, p = 0, 𝜂G2 = 0.999). The interaction
between treatment and storage period (F(70, 270) = 2800, p = 0,
𝜂G2 = 0.999) also was significant on FFA. In the genotype HHB67,
the effect of treatment (F(14, 270) = 66,383, p = 0, 𝜂G2 = 1),
storage period (F(5, 270) = 171,770, p = 0, 𝜂G2 = 1), and that
of the interaction between treatment and storage period (F(70,
270) = 10,494, p = 0, 𝜂G2 = 1) were significant. Similarly, the effect of
treatment (F(14, 270) = 161,263, p = 0, 𝜂G2 = 1), storage period (F(5,
270) = 155,038, p = 0, 𝜂G2 = 1), and that of the interaction between
treatment and storage period (F(70, 270) = 7000, p = 0, 𝜂G2 = 0.999)
were significant.
Heat treatment of flour at higher temperature inactivates the li-
pase enzyme thereby preventing the release of FFA (Gili et al., 2018;
Mazaheri et al., 2019; Ruge et al., 2012; Xu et al., 2016). Heating dis-
rupts non-covalent bonds between amino acid residues resulting in re-
versible loss of enzyme activity. Prolonged heating permanently inac-
tivates the enzyme (Sawai et al., 2003). A reduction in FFA of flour
(63.6%) was observed when pearl millet grains were treated at 100 °C
for 120 min (Bhati et al., 2016). The FFA values of pearl millet flour
stored in different packaging materials after thermal treatment were
significantly different than untreated flour on same day of analysis.
Hydrolytic changes due to action of lipolytic enzymes produce high
FFA value. Further, continous action of lipase during storage leads
to higher FFA value in control than treated samples (Yadav et al.,
2012b).
3.1.2. Effect on alcoholic acidity (AA)
Overall effect of treatment, storage period and genotype on AA
showed significant variations between genotypes (df=2, p < 2.22e-16,
sig.0.001), between treatments (df=14, p < 2.22e-16, sig.0.001), and
between storage period (df=5, p < 2.22e-16, sig.0.001). The interac-
tion effects between genotypes and treatments (df=28, p < 2.22e-16,
sig.0.001), between genotypes and storage period (df=10, p < 2.22e-
16, sig.0.001), between treatments and storage period (df=70,
p < 2.22e-16, sig.0.001), and between genotypes, treatments and stor-
age period (df=140, p < 2.22e-16, sig.0.001) were all significant
for AA.
The influence of treatment and storage period on the rancidity pa-
rameter AA in each genotype was analysed with two-way ANOVA.
In the genotype Dhanshakthi, AA showed significant variations due
to the effect factors viz., treatment (F(14, 270) = 55.8, p = 3.48e-
71, 𝜂G2 = 0.743), and storage period (F(5, 270) = 392, p = 1.64e-
12, 𝜂G2 = 0.879). The interaction between treatment and storage
period (F(70, 270) = 6.40, p = 3.27e-29, 𝜂G2 = 0.624) also was
significant on AA. In the genotype HHB67, the effect of treatment
(F(14, 270) = 190, p = 9.79e-13, 𝜂G2 = 0.908), storage period (F(5,
270) = 1592, p = 5.00e-19, 𝜂G2 = 0.967), and that of the interaction
between treatment and storage period (F(70, 270) = 19.6, p = 1.88e-73,
𝜂G2 = 0.836) were significant. In the genotype MPMH21, the effect of
treatment (F(14, 270) = 563, p = 1.29e-19, 𝜂G2 = 0.967), storage period
(F(5, 270) = 1290, p = 3.99e-18, 𝜂G2 = 0.96), and that of the interaction
4
Food Chemistry Advances 2 (2023) 100190
between treatment and storage period (F(70, 270) = 37.3, p = 3.54e-10,
𝜂G2 = 0.906) were significant.
Among the treatments, thermal treatment is significantly different
from other non thermal treatments. The lower AA values initially in ther-
mal treated samples were due to inhibition of lipase by thermal treat-
ment. The AA of untreated and heat treated samples of HHB 67 increased
significantly during storage from 0.23 to 1.00 g.100 g−1 of dry matter
and from 0.19 to 0.66 g.100 g−1 of dry matter respectively. This is due
to increase of free fatty acids by enzymatic lipolysis. Fivefold increase
in fat acidity was observed in the flour of control and hot‐air treated
pearl millet grain, while it remained same in the flour of the boiled
grain during storage under ambient conditions for a month (Chavan &
Kachare, 1994).
3.1.3. Effect on lipase activity (LA)
LA was significantly varied between genotypes (df=2, p < 2e-16,
sig.0.001), between treatments (df=14, p < 2e-16, sig.0.001), and
between storage period (df=5, p < 2e-16, sig.0.001). The interac-
tion effects between genotypes and treatments (df=28, p < 2e-16,
sig.0.001), between genotypes and storage period (df=10, p < 2e-
16, sig.0.001), between treatments and storage period (df=70,
p < 2e-16, sig.0.001), and between genotypes, treatments and stor-
age period (df=140, p < 2e-16, sig.0.001) were all significant
for LA.
The influence of treatment and storage period on the rancid-
ity parameter LA in each genotype was analysed with two-way
ANOVA. In the genotype Dhanshakthi, the effect of treatment (F(14,
270) = 398, p = 4.14e-17, 𝜂G2 = 0.954), storage period (F(5,
270) = 1093, p = 7.77e-17, 𝜂G2 = 0.953), and that of the interaction
between treatment and storage period (F(70, 270) = 44.4, p = 2.27e-
11, 𝜂G2 = 0.92) were significant. In the genotype HHB67, the ef-
fect of treatment (F(14, 270) = 586, p = 6.96e-19, 𝜂G2 = 0.968),
storage period (F(5, 270) = 1292, p = 3.14e-18, 𝜂G2 = 0.96), and
that of the interaction between treatment and storage period (F(70,
270) = 26.1, p = 3.00e-87, 𝜂G2 = 0.871) were significant. In the geno-
type MPMH21, the effect of treatment (F(14, 270) = 971, p = 1.07e-
22, 𝜂G2 = 0.981), storage period (F(5, 270) = 1452, p = 8.54e-19,
𝜂G2 = 0.964), and that of the interaction between treatment and storage
period (F(70, 270) = 80.6, p = 1.23e-14, 𝜂G2 = 0.954) were significant
on LA.
Among the treatments, thermal treatment is significantly different
from other non thermal treatments. Thermal treatment causes denat-
uration of the enzymes and decreases the activity of lipase. Milling
process rapidly deteriorates lipids in the bran by lipase and lipoxyge-
nase (LOX) rendering the flour unacceptable for consumption (Zhang &
Hamaker, 2005). Fatty acids at 1, 3-site of triglycerides are specifically
cleaved by lipase and the released free fatty acids are further oxidized
by lipoxygenase and atmospheric oxygen leading to peroxide formation.
Yadav et al., 2012a reported decrease in lipase activity of pearl millet
grains on application of microwave heating.
Similarly, heat processing of proso millets with steam-jacketed
paddle conveyor at 110 °C completely inactivated lipolytic enzymes
(Bookwalter et al., 1987). Moreover, 96% reduction of lipase activity
during steam heating was observed in whole wheat flour (Rose et al.,
2008). Lipase inactivation was also observed in naked oat kernels
(Keying et al., 2009).
3.1.4. Effect on peroxide value (PV)
PV showed significant variations between genotypes (df=2,
p < 2.22e-16, sig.0.001), between treatments (df=14, p < 2.22e-16,
sig.0.001), and between storage period (df=5, p < 2.22e-16, sig.0.001).
The interaction effects between genotypes and treatments (df=28,
p < 2.22e-16, sig.0.001), between genotypes and storage period (df=10,
p < 2.22e-16, sig.0.001), between treatments and storage period (df=70,
p < 2.22e-16, sig.0.001), and between genotypes, treatments and stor-
age period (df=140, p < 2.22e-16, sig.0.001) were all significant for PV.
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al. Food Chemistry Advances 2 (2023) 100190

Fig. 1. Significance between interquartile ranges of treatment effects on (A) FFA, (B) PV, (C) AA, and (D) LA in pearl millet flour.

The influence of treatment and storage period on the ran- 3.2. Within comparison of treatments, storage periods, and genotypes for
cidity parameter PV in each genotype was tested with two-way rancidity parameters
ANOVA. In the genotype Dhanshakthi, the effect of treatment (F(14,
270) = 68.5, p = 3.14e-80, 𝜂G2 = 0.78), storage period (F(5, Effect of genotypes, treatments and storage periods individually on
270) = 978, p = 1.25e-17, 𝜂G2 = 0.948), and that of the interaction rancidity parameters are presented below.
between treatment and storage period (F(70, 270) = 4.82, p = 3.94e-
21, 𝜂G2 = 0.556) were significant. In the genotype HHB67, the effect of
treatment (F(14, 270) = 153, p = 2.66e-11, 𝜂G2 = 0.888), storage period
3.2.1. Comparison of treatments
(F(5, 270) = 513, p = 1.29e-13, 𝜂G2 = 0.905), and that of the interaction
The variability between individual treatments for rancidity parame-
between treatment and storage period (F(70, 270) = 13.6, p = 3.95e-57,
ters were tested using interquartile ranges of FFA, AA, LA, and PV. The
𝜂G2 = 0.779) were significant on PV. In the genotype MPMH21, the
results are presented in the form of bar graphs with alphabetical signifi-
effect of treatment (F(14, 270) = 138, p = 5.71e-11, 𝜂G2 = 0.877),
cance notations (Fig. 1). All the treatments were statistically significant
storage period (F(5, 270) = 1444, p = 1.69e-19, 𝜂G2 = 0.964), and
from each other for FFA Fig. 1(A). In all the three genotypes, the com-
that of the interaction between treatment and storage period (F(70,
bined treatment of Lmn_Th_N had decreased the FFA value by 10–15
270) = 9.04, p = 5.98e-41, 𝜂G2 = 0.701) were significant.
fold compared to control at 60 days of storage. PV was statistically on
Peroxide formation is mainly due to oxidative rancidity. As ther-
par with the following combinations: LDPE_N and Lmn; Refrn, LDPE_Th,
mal treatment has no effect directly on peroxide formation, perox-
and HDPE_N; HDPE_Th and Lmn_N; Lmn_Th and LDPE_Th_N; while sig-
ide value has not significantly reduced compared to packaging mate-
nificantly different for all other treatments Fig. 1(B). In all the three
rial. Thermal treatment showed a significant lower PV than untreated
genotypes, the combined treatment of Lmn_Th_N had decreased the PV
samples. The lower amounts of hydroperoxides formed was possibly
value by 2 fold compared to control at 60 days of storage. Similarly, AA
due to inactivation of lipolytic enzymes, resulting decrease in forma-
was statistically on par for the following combinations: LDPE and HDPE;
tion of free fatty acids and exclusion of oxygen by packaging material
Lmn and LDPE_N; Ctrl_Th and Refrn; LDPE_Th, HDPE_T, and Lmn_N;
(Eskin & Przybylski, 2001). Maillard reaction products produced dur-
HDPE_T_N and Lmn_Th_N; while showing statistical significance for rest
ing toasting scavenge radicals and prevent formation of hydroperoxides
of the treatments Fig. 1(C). The combination treatment of Lmn_Th_N
(Yoshimura et al., 1997). Moreover, low amounts of peroxides in pro-
had decreased the AA value by 2–3 fold compared to control at 60 days
longed storage are due to decomposition of unstable hydroperoxides
of storage. Similar to the FFA, the LA was significantly different for all
into secondary oxidation products (Eskin & Przybylski, 2001).
the treatments, but for Lmn and LDPE_N which were on par Fig. 1(D).

5
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
Fig. 2. Significance between interquartile ranges of storage period effects on (A) FFA; (B) PV; (C) AA; (D) LA in pearl millet flour.
Dry heat treatment of pearl millet grains was found to retard lipid
degradation in the flour during storage (Arora et al., 2002; Kadlag et al.,
1995; Patel & Parameswaran, 1992). Hydrothermal treatment of pearl
millet grain has extended the shelf-life of flour up to 50 days at am-
bient conditions (Yadav et al., 2012a). A significant decrease was re-
ported in activities of lipase (47.8%), lipoxygenase (84.8%), peroxi-
dase (98.1%) and polyphenol oxidase (100%) in HT-HTh-thNIR (hydro
treated- hydrothermal- thermal near infrared rays) treated flour com-
pared to the individual treatments. Moreover decrease of 67.84% and
66.4% of free fatty acid and peroxide contents were observed in flour
without altering starch and protein digestibility (Vinutha et al., 2022).
3.2.2. Comparison of storage periods
The rancidity parameters at different days of storage were statisti-
cally compared using interquartile ranges of readings (Fig. 2). The days
of storage at six intervals were significantly different for FFA, PV, and
AA. Further, the duration of storage of pearl millet flour had no signifi-
cant effect on lipase activity till 7th day and likewise, the storage period
from 15th day to 30th day were on par.
Thiam (1977) reported a high FFA value due to hydrolytic
changes associated with the action of lipolytic enzymes. Further,
continuous lipase activity during storage may have lead to a sig-
nificantly higher FFA value in control than treated flour. Peroxide
value increased significantly in control flour after 30 days of stor-
age (Yadav et al., 2012a). The increasing trend of PV during storage
was also in agreement with the observation of Chaudhary and Kapoor
(1984).
3.2.3. Comparison of genotypes
The genotypic variability among the three tested cultivars of pearl
millet was statistically compared using interquartile ranges for rancid-
6
Food Chemistry Advances 2 (2023) 100190
ity parameters. Flours from all three pearl millet genotypes, HHB 67,
MPMH 21 and Dhanshakthi were significantly different for FFA, PV,
AA, and LA (Fig. 3).
The trends in rate of change in rancidity parameters during the pe-
riod of storage in flours of three genotypes are shown in the Fig. 4. The
rate of change of FFA in all the three genotypes increased in the first
week, and gradually decreased in Dhanshakthi. HHB 67 and MPMH 21
showed high and sharp increase in FFA in the first week, and steeply de-
creased thereafter Fig. 4(A). All the three genotypes had irregular rise
and fall in AA during the test storage period of two months Fig. 4(B). The
flour of genotype MPMH 21 produced slight and gradual increase in LA
upto 30 days and gradually decreased thereafter. Dhanshakthi showed
alternate increase and decrease at every fortnight on a decreasing trend
for LA. The genotype HHB 67 showed alternate increase and decreasing
trend in LA with highest rate of increase from 30 to 45 days of storage
Fig. 4(C). The genotypes HHB 67 and MPMH 21 showed gradual de-
crease in PV after one week of storage. Dhanshakthi showed very high
and steep increase at 7th day and had a steep decrease at 30th day in rate
of change in PV which gradually decreased thereafter Fig. 4(D). Perox-
ide values were found to increase till 10 to 17 days in various genotypes
of pearl millet (Mazumdar et al., 2016). Untreated flour had unpleas-
ant off odor and bitter taste at the 10th day of storage (Yadav et al.,
2012a).
3.3. Role of rancidity parameters in groupings of intervention effects
(treatments, storage periods, and genotypes)
The data was analyzed to understand the rancidity factors contribut-
ing to the variability using Principal Component Analysis.
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al. Food Chemistry Advances 2 (2023) 100190

Fig. 3. Significance between interquartile ranges of genotype effects on (A) FFA; (B) PV; (C) AA; (D) LA in pearl millet flour.

7
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
Fig. 4. (A) Rate of change in free fatty acids of pearl millet genotypes upon storage; (B) Rate of change in alcoholic acidity of pearl millet genotypes upon storage;
(C) Rate of change in lipase activity of pearl millet genotypes upon storage; (D) Rate of change in peroxide value of pearl millet genotypes upon storage.
3.3.1. Important contributing factors (rancidity parameters) to the
treatment variability
PCA showed that the PC1 and PC2 explained 77.6% and 12.7% vari-
ability respectively. PCA grouping of treatments with linear projection
of response variables showed that PV contributed the highest to variabil-
ity followed by AA and FFA. There was a positive correlation existing
between FFA and LA Fig. 5(A). The plot reveals a loading trend with
half the number of treatments in the direction of PV and the remaining
half in the direction of LA and FFA.
3.3.2. Important contributing factors (rancidity parameters) to the storage
time period variability
Upto a week, there was not much activity of LA, PV, FFA, AA. With
the progress of days of storage the contribution of rancidity parameters
PV, FFA, and LA increased Fig. 5(B). This clearly shows the shelf life of
flour starts deteriorating after a week of storage.
3.3.3. Important contributing factors (rancidity parameters) to the
genotype variability
Among the three genotypes, Dhanshakthi showed strong inclina-
tion towards PV and other two genotypes HHB 67 and MPMH 17
scoring for other variables viz. FFA, AA and LA. Fig. 5(C). The role
of PV in the storability of Dhanshakthi is an important factor to be
considered.
8
Food Chemistry Advances 2 (2023) 100190
3.4. K-means clustering of treatments to find out important treatments that
are useful to increase the shelf life of pearl millet flour
The data was also analyzed for K-means clustering of treatments to
find out important treatments that are useful to increase the shelf life of
pearl millet flour.
Treatments are clustered into three groups using k-means cluster-
ing (Fig. 6). All treatments involving thermal conditioning are grouped
separately (cluster 1) from other non-thermal treatments (cluster 3),
while control remained as a separate group (cluster 2). It indicates exis-
tence of two trends: first, a clear role of thermal conditioning in altering
rancidity parameters in pearl millet flour during storage and second,
all the tested intervention measures of treatments and storage periods
having influence on rancidity. PV emerged as a single most important
parameter responsible for increased rancidity in control. FFA followed
by AA and LA are major parameters contributing to rancidity levels
in non thermal treatments. Treatment LDPE had greater influence on
LA and AA while, HDPE, LDPE_N, HDPE_N, Lmn, Lmn_N had influence
on FFA.
The Lmn_Th_N is the best treatment followed by Lmn_Th, HDPE_Th,
LDPE_Th, HDPE_Th_N, LDPE_Th_N for increasing the shelf life of pearl
millet flour as it is away from the direction of linear projection of vari-
ables AA, FFA, LA and PV. In a similar way, it was found that different
packaging and storage interventions influenced shelf life of yam flour
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
Fig. 5. (A) PCA grouping of treatments with linear projections of FFA, LA, AA,
PV; (B) PCA grouping of days of storage with linear projections of FFA, LA, AA,
PV; (C) PCA grouping of genotypes with linear projections of FFA, LA, AA, PV.
(Adebowale et al., 2017) and wheat flour (Majzoobi et al., 2011). In
finger millet, processing interventions significantly improved the func-
tional properties of flours (Navyashree et al., 2022).
4. Conclusion
Although pearl millet grain is storable for months, milled flour has
very short shelf life of 5 to 7 days due to high fat content, and lipolytic
action of enzymes. Lipolytic enzymes which were sequestered in com-
partments of cells in whole grain get access to lipids when milled to
flour. This leads to release of free fatty acids which are substrates for hy-
drolytic degradation by enzymes or oxidative damage by atmospheric
oxygen leading to peroxide formation which is the major cause of off
flavours and bitterness. In the present study, thermal treatment of grains
9
Food Chemistry Advances 2 (2023) 100190
Fig. 6. K-means clustering of treatments with linear projections of variables.
at 150 °C for 30 min before milling and storage in laminated pouches un-
der nitrogen atmosphere was found to extend shelf life by 60 days com-
pared to untreated samples stored in LDPE and HDPE packaging under
atmospheric air. Genotypes, treatments and storage period had signifi-
cant effect individually and interactively on all rancidity parameters like
lipase activity, FFA, alcoholic acidity and peroxide value compared to
control. Heat treatment inactivates enzymes and inactivation of lipase
leads to reduced formation of free fatty acids. Low FFA and modified
atmosphere further reduce formation of peroxides. Thermal treatment
had resulted in low AA, FFA throughout the storage period compared
to control. This integrated approach of heat treatment and storage in
laminated pouches under nitrogen atmosphere decreased FFA value by
10–15 fold, peroxide value by 2 fold and AA by 2–3 fold and extended
the shelf life of pearl millet flour by 60 days. K-means clustering has
clearly indicated the prominent role of thermal treatment and also the
influence of genotype and packaging on rancidity parameters in shelf
life extension.
Funding
This work was financially supported by the National Food Security
Mission (NFSM) Department of Agriculture, Cooperation and Farmers
Welfare (DAC & FW), New Delhi, India.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
CRediT authorship contribution statement
P.G. Padmaja: Conceptualization, Methodology, Formal anal-
ysis, Investigation, Writing – original draft, Funding acquisi-
tion. A. Kalaisekar: Formal analysis, Writing – original draft. R.
Venkateswarlu: Methodology, Investigation, Writing – original draft.
S. Shwetha: Investigation. B. Dayakar Rao: Writing – original draft,
P.G. Padmaja, A. Kalaisekar, R. Venkateswarlu et al.
Funding acquisition. V.A. Tonapi: Writing – original draft, Funding
acquisition.
Data Availability
Data will be made available on request.
Acknowledgments
Indian Council of Agricultural Research (ICAR), New Delhi, and
ICAR- Indian Institute of Millets Research (ICAR-IIMR) are sincerely ac-
knowledged for the facilities provided during the study.
Supplementary materials
Supplementary material associated with this article can be found, in
the online version, at doi:10.1016/j.focha.2023.100190.
References
Abdalgader, M. A. A., Ashraf, S. A., & Awadelkareem, A. M. (2019). Effect of natural and
synthetic antioxidant on shelf-life of different sudanese Pennisetum glaucum L. flours.
Bioscience Biotechnology Research Communications, 12(3), 652–657.
Adebowale, A. A., Owo, H. O., Sobukola, O. P., Obadina, O. A., Kajihausa, O. E., Ade-
gunwa, M. O., et al., (2017). Influence of storage conditions and packaging materials
on some quality attributes of water yam flour. Cogent Food Agriculture, 3(1), Article
1385130. doi:10.1080/23311932.2017.1385130.
Altman, N., & Krzywinski, M. (2015). Split plot design. Nature Methods, 12, 165–166.
doi:10.1038/nmeth.3293.
Arora, P., Sehgal, S., & Kawatra, A. (2002). The role of dry heat treatment in improving
the shelf life of pearl millet flour. Nutrition and Health, 16, 331–336.
Bhati, D., Bhatnagar, V., & Acharya, V. (2016). Effect of pre-milling processing techniques
on pearl millet grains with special reference to iron availability. Asian Journal of Dairy
and Food Research, 35(1), 76–80.
Bookwalter, G. N., Lyle, S. A., & Warner, K. (1987). Millet processing for improved sta-
bility and nutritional quality without functional changes. Journal of Food Science, 52,
399–402.
Chavan, J. K., & Kachare, D. P. (1994). Effects of seed treatment on lipolytic deterioration
of pearl millet flour during storage. Journal of Food Science and Technology, 31(1),
80–81.
Chaudhary, P., & Kapoor, A. C. (1984). Changes in the nutritional value of pearl millet
flour during storage. Journal of the Science of Food and Agriculture, 35, 1219–1224.
doi:10.1002/jsfa.2740351113.
Chinenye, O. E., Ayodeji, O. A., & Baba, A. J. (2017). Effect of fermentation (natural and
starter) on the physicochemical, anti-nutritional and proximate composition of pearl
millet used for flour production. American Journal of Bioscience and Bioengineering, 5,
12–16.
Demsar, J., Curk, T., Erjavec, A., Zupan, B., et al., (2013). Orange: Data mining tool box
in Python. Journal of Machine Learning Research, 14, 2349–2353.
Directorate of Millets Development. (2020). Department of agriculture, cooperation & farmers
welfare, ministry of agriculture & farmers welfare. Government of India.
Eskin, N. A. M., Przybylski, R., Eskin, N. A. M., & Robinson, D. S. (2001). Antioxidants and
shelf life of foods. In Food shelf life stability: Chemical, biochemical, and microbiological
changes (pp. 175–209). Boca Raton, FL: CRC Press.
Galliard, T., Allen, J. C., & Hamilton, R. J. (1999). Rancidity in cereal products. In Rancidity
in foods (pp. 140–156). Gaithersburg, Maryland: Aspen Publishers.
Gili, R. D., Penci, M. C., Torrez Irigoyen, R. M., Giner, S. A., & Ribotta, P. D. (2018). Effect
of wheat germ heat treatment by fluidised bed on the kinetics of lipase inactivation.
Food and Bioprocess Technology, 11, 1002–1011.
Gomez, M. I., Gupta, S. C., & Caballero, B. (2003). Millets. In Encyclo-
pedia of food sciences and nutrition (pp. 3974–3979). Academic Press.
doi:10.1016/B0-12-227055-X/00791-4.
Goswami, S., Asrani, P., Ansheef Ali, T. P., Kumar, R. D., Vinutha, T., Veda, K., et al.,
(2020). Rancidity matrix: Development of biochemical indicators for analysing the
keeping quality of pearl millet flour. Food Analytical Methods, 13, 2147–2164.
doi:10.1007/s12161-020-01831-2.
Hara, A., & Radin, N. S. (1978). Lipid extraction of tissue with low toxicity solvent. Ana-
lytical Biochemistry, 90(1), 420–426. doi:10.1016/0003-2697(78)90046-5.
Hintlian, C. B., & Hotchkiss, J. H. (1986). The safety of modified atmosphere packaging:
A review. Food Technology, 40(12), 70–76.
IS 12711. (1989). Method of determination of alcoholic acidity. New Delhi: Bureau of Indian
Standards.
Kadlag, R. V., Chavan, J. K., & Kachare, D. P. (1995). Effects of seed treatments and
storage on the changes in lipids of pearl millet meal. Plant Foods for Human Nutrition,
47, 279–285.
Kapoor, R., & Kapoor, A. C. (1990). Effect of different treatments on keeping quality of
pearl millet flour. Food Chemistry, 35, 277–286.
10
Food Chemistry Advances 2 (2023) 100190
Keying, Q., Changzhong, R., & Zaigui, L. (2009). An investigation on pretreatments for
inactivation of lipase in naked oat kernels using microwave heating. Journal of Food
Engineering, 95, 280–284. doi:10.1016/j.jfoodeng.2009.05.002.
Kwon, D. Y., & Rhee, J. S. (1986). A simple and rapid colorimetric method for determina-
tion of free fatty acids for lipase assay. Journal of American Oil Chemists’ Society, 63,
89–92.
Longvah, T., Ananthan, R., Bhaskarachary, K., & Venkaiah, K. (2017). Indian food com-
position tables. Hyderabad: National Institute of Nutrition, Indian Council of Medical
Research.
Majzoobi, M., Farahnaky, A., & Agah, S. (2011). Properties and shelf-life of part-and ful-
l-baked flat bread ‘barbari’ at ambient and frozen storage. Journal of Agricultural Sci-
ence and Technology, 13(6), 1077–1090.
Mazaheri, Y., Torbati, M., Damirchi, S. A., & Savage, G. P. (2019). Effect of roasting and
microwave pre-treatments of Nigella sativa L. seeds on lipase activity and the quality
of the oil. Food Chemistry, 274, 480–486. doi:10.1016/j.foodchem.2018.09.001.
Mazumdar, S. D., Gupta, S. K., Banerjee, R., Gite, S., Durgalla, P., & Bagade, P. (2016).
Determination of variability in rancidity profile of select commercial Pearl millet va-
rieties/hybrids. In Proceedings of the CGIAR research program on dryland cereals review
meeting October 5-6, 2016.
Mohajerkhorasani, S., Alami, M., Kashaninejad, M., & Shahiritabarestani, H. (2019). In-
creasing the shelf-life of millet flour by using heat-moisture and microwave treat-
ments. Iranian Journal of Food Science and Technology, 16(86), 83–93.
ChapterWiley (Eds.) Mullan, M., McDowell, D., Coles, R., & Kirwan, M. (2011). Modified
atmosphere packaging. In Food and beverage packaging technology edition (pp. 263–294)
Modified Atmosphere Packaging Publisher. ChapterWiley (Eds.).
Nantanga, K.K.M. (2006). Lipid stabilization and partial pre-cooking of pearl
millet by thermal treatments (12-105) MSc (Agric.)Thesis in Food Science
and Technology: Department of FoodScience, Faculty of Natural and Agri-
cultural Sciences, University of Pretoria. https://repository.up.ac.za/bitstream/
handle/2263/26680/dissertation.pdf?sequence=1.
Navyashree, N., Sengar, A. S., Sunil, C. K., & Venkatachalapathy, N. (2022).
White finger millet (KMR-340): A comparative study to determine the effect
of processing and their characterization. Food Chemistry, 374, Article 131665.
doi:10.1016/j.foodchem.2021.131665.
Ogori, A. F., Jatua, M. K., & Adamu, L. A. U. M. (2013). Effect of semi-densed polyethylene
storage on organoleptic and chemical characteristics of flours from soaked, malted
and their blend of millet grains (Pennesitum glacum). American Journal of Biological,
Chemical and Pharmaceutical Sciences, 1(7), 81–88.
Patel, K. V., & Parameswaran, M. (1992). Effect of heat treatment on lipid degradation in
bajra flour. Journal of Food Science and Technology, 29, 51–52.
Malviya, P. K. (2015). Shelf-life enhancement of pearl millet flour. Journal of Food Pro-
cessing Technology, 6, 10.
Raheem, D., Dayoub, M., Birech, R., & Nakiyemba, A. (2021). The contribution of ce-
real grains to food security and sustainability in Africa: Potential application of UAV
in Ghana, Nigeria, Uganda, and Namibia. Urban Science, 5, 8. doi:10.3390/urban-
sci5010008.
Reddy, V. P., Faubion, J. M., & Hoseney, R. C. (1986). Odor generation in ground, stored
pearl millet. Cereal Chemistry, 63, 403–406.
Rose, D. J., Ogden, L. V., Dunn, M. L., & Pike1, O. A. (2008). Enhanced lipid stability in
whole wheat flour by lipase inactivation and antioxidant retention. Cereal Chemistry,
85(2), 218–223.
Ruge, C., Changzhong, R., & Zaigui, L. (2012). The effects of different inactivation treat-
ments on the storage properties and sensory quality of naked Oat. Food and Bioprocess
Technology, 5, 1853–1859. doi:10.1007/s11947-011-0551-5.
Sawai, J., Sagara, K., Hashimoto, A., Igarashi, H., & Shimizu, M. (2003). Inactivation char-
acteristics shown by enzymes and bacteria treated with far-infrared radiative heating.
International Journal of Food Science and Technology, 38, 661–667.
Serba, D. D., Yadav, R. S., Varshney, R. K., Gupta, S. K., Mahalingam, G., Srivastava, R. K.,
Kole, C., et al., (2020). Genomic designing of pearl millet: A resilient crop for arid and
semi-arid environments. In Genomic designing of climate-smart cereal crops (pp. 221–
286). Cham: Springer. doi:10.1007/978-3-319-93381-8_6.
Thiam, A. A. (1977). Contribution to the study of the biochemical phenomena of millet and
sorghum flour determination. In Proceedings of the tropical products institute conference
papers. Institut Technologie Alimentaire (p. 69).
Varsha, R., & Narayanan, A. (2017). Storage stability of biofortified pearl millet flour.
International Journal of Agriculture Innovations and Research, 5(5), 709–713.
Vinutha, T., Kumar, D., Bansal, N., Krishnan, V., Goswami, S., Kumar, R. R., et al., (2022).
Thermal treatments reduce rancidity and modulate structural and digestive properties
of starch in pearl millet flour. International Journal of Biological Macromolecules, 195,
207–216. doi:10.1016/j.ijbiomac.2021.12.011.
Xu, B., Wang, L. K., Miao, W. J., Wu, Q. F., Liu, Y. X., Sun, Y., et al., (2016). Thermal
versus microwave inactivation kinetics of lipase and lipoxygenase from wheat germ.
Journal of Food Process Engineering, 39(3), 247–255. doi:10.1111/jfpe.12216.
Yadav, D. N., Anand, T., Kaur, J., et al., (2012a). Improved storage stability of pearl millet
flour through microwave treatment. Agricultural Research, 1, 399–404.
Yadav, D. N., Kaur, J., Anand, T., & Singh, A. K. (2012b). Storage stability and pasting
properties of hydrothermally treated pearl millet flour. International Journal of Food
Science and Technology, 47, 2532–2537. doi:10.1111/j.1365-2621.2012.03131.x.
Yoshimura, Y., Iijima, T., Watanabe, T., & Nakazawa, H. (1997). Antioxidative effect of
Maillard reaction products using glucose–glycine model system. Journal of Agriculture
and Food Chemistry, 45, 4106–4109.
Zhang, G., & Hamakar, B. R. (2005). Sorghum (Sorghum bicolor L. Moench) flour pasting
properties influenced by free fatty acids and protein. Cereal Chemistry, 82(5), 534–540.
doi:10.1094/CC-82-0534.