Food Bioscience 45 (2022) 101508

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Food Bioscience
journal homepage: www.elsevier.com/locate/fbio

Development of a mixed fruit beverage and pulsed light treatment thereof

to obtain a microbially safe and enzymatically stable product
Snehasis Chakraborty *, Sagar Mahale, Rishab Dhar, Somnath Basak
Food Engineering and Technology Department, Institute of Chemical Technology Matunga (E), Mumbai, 400 019, India

A R T I C L E I N F O A B S T R A C T

Keywords: A mixed fruit beverage was formulated according to D-optimal mixture design using apple ber juice (X1, 45–59%
D-optimal mixture design v/v) - an underexplored fruit, carambola juice (X2, 1–20% v/v), and black table grape juice (X3, 35–40% v/v).
Numerical optimization The crisp overall acceptability (COA) from 16 semi-trained panelists and overall hedonic score (OSH) from
Non-thermal processing
consumers were estimated. The best formulation was apple ber: carambola: black table grape juices = 59.0: 3.5:
Pasteurization
Microbial safety
37.5 v/v with COA and OSH values of 3.23 (on a scale of 5) and 7.55 (on a scale of 9), respectively. Further, the
Enzymatic stability efficacy of pulsed light (PL) (18–30 W cm− 2 for 33–167 s; total fluence 600–5000 J cm− 2) and thermal treatments
(90 ◦ C for 1–5 min) were studied on various attributes of the mixed beverage. The beverages processed at 90 ◦ C
for 5 min (thermal treatment) and 5000 J cm− 2 (PL treatment at 30 W cm− 2 for 167 s) showed no microbial count
with a complete enzyme inactivation. The thermally pasteurized sample (90 ◦ C/5 min) was more brown
(browning index = 70.15) than the PL pasteurized beverage (browning index = 65.98). Interestingly, the PL
pasteurized (5000 J cm− 2) beverage showed 25%, 27%, and 19% more vitamin C, antioxidants, and phenolic
compounds, respectively, than the thermally pasteurized (90 ◦ C/5 min) beverage. The PL-treated beverage had a
greater sensory appeal than the thermally treated beverage. PL-treatment can be an efficient non-thermal
pasteurization treatment for ensuring complete microbial and enzymatic inactivation with maximal retention
of nutritional and sensory attributes of the blended beverage.

1. Introduction
India ranks second in the world in total fruit production, but only
2–3% of this is converted to processed products (National Horticulture
Board, 2019). Apart from the major fruits such as grapes, pineapple,
pomegranates, mangoes, bananas, and oranges, there is a long list of
underutilized fruits produced across the country. Underutilized means
that the fruit variety is locally grown and remains underexplored besides
having a limited shelf-life. The difficulties faced during post-harvest
operations also make these fruits underutilized and restrict their com­
mercial uses (Monika, 2014).
Apple ber (Zizyphus mauritiana), also referred to as Indian jujube, is
one of the underutilized fruits widely grown in India. The juice from
apple ber is an excellent source of vitamin C (500–600 mg/L) (Füzfai
et al., 2004). Carambola (Averrhoa carambola) also comes under the
underutilized fruit category depending upon its commercial use in India
to date. Carambola, also locally recognized as star fruit, is rich in phy­
tochemicals, vitamins, minerals like potassium, and organic acids,
mainly oxalic acid. Its use in reducing hyperglycemia and hyperlipid­
emia is noteworthy (Shui & Leong, 2006). Apple ber and carambola
have a history of safe consumption in India and are consumed locally
due to their pharmacological benefits (Prakash et al., 2021; Sharma
et al., 2015). The black table grape is an excellent source of phenolics
and antioxidants, mainly anthocyanins, flavonoids, and resveratrol
(Rolle et al., 2013). Today’s consumer demand for nutritionally
balanced fruit juice. Therefore, a blend of these three fruit juices may
have the potential to fulfill that demand.
Developing a fruit juice blend has been exercised by many re­
searchers, including some tropical fruits (Jaeger et al., 2009). Moreover,
in most of the studies, the formulation was optimized based on the
sensory acceptance of the same. The examples include
pomegranate-amla-muskmelon based mixed beverage (Bhalerao et al.,
2020), amla-pineapple-coconut water (Dhar, Bhalerao, et al., 2021),
litchi based mixed fruit beverage (Jayachandran, Chakraborty, & Rao,
2015a, 2015b), mixed fruit squash (Jothi et al., 2014), and mixed berry
juices (Schiassi et al., 2020). However, optimization of the formulation

* Corresponding author.
E-mail addresses: sc.chakraborty@ictmumbai.edu.in (S. Chakraborty), sammahale193@gmail.com (S. Mahale), rishabdhar94@gmail.com (R. Dhar), somnath.
basak17@gmail.com (S. Basak).

https://doi.org/10.1016/j.fbio.2021.101508
Received 26 June 2021; Received in revised form 3 November 2021; Accepted 12 December 2021
Available online 14 December 2021
2212-4292/© 2021 Elsevier Ltd. All rights reserved.
S. Chakraborty et al. Food Bioscience 45 (2022) 101508

taking apple ber, carambola, and table grape juices has not been Table 1
attempted. Crisp overall acceptability (COA), overall hedonic scores (OSH), and mixture
The primary concern of the mixed fruit juice processing industry is its components for each formulation obtained from D-optimal mixture design.
limited shelf-life (Lan et al., 2021). A 5-log reduction in the pertinent Sample Juice components in 100 mL Overall Crisp overall
microorganism has been recommended for fruit juice pasteurization No. mixture hedonic acceptability
(Lima Tribst et al., 2009). The thermal treatment at 80–95 ◦ C for 1–10 score (OSH) (COA)
Apple Carambola Grape
min is generally applied for fruit juice pasteurization. However, the ber (mL) (mL)
negative impact of intense heat treatment on the bioactive compounds (mL)
encourages the exploration of an alternative technique for the juices S1 59.0 3.5 37.5 7.80 ± 3.29 ± 0.05f
(Chakraborty et al., 2016). Pulsed light (PL) is a non-thermal technique 0.12e
that involves the exposure of the food under a compound spectrum S2 59.0 3.5 37.5 7.77 ± 3.20 ± 0.04f
0.17e
(wavelength of 100–1100 nm) of UV, visible and infrared counterparts S3 55.6 4.4 40.0 7.64 ± 2.98 ± 0.02e
(Xiaokang et al., 2020). PL technique has been applied in various fruit 0.25e
juices (Mahendran et al., 2019; Dhar, Basak, et al., 2021). The juice S4 55.6 4.4 40.0 7.82 ± 2.86 ± 0.03d
matrices tested under PL exposure were amla-pineapple-coconut 0.32e
S5 56.8 5.8 37.4 6.79 ± 2.81 ± 0.02d
water-based mixed fruit beverage (Dhar & Chakraborty, 2020), pine­
0.34d
apple juice (Vollmer et al., 2020), amla juice (Chakraborty et al., 2020), S6 59.0 6.0 35.0 7.17 ± 3.06 ± 0.03e
apple juice (Muñoz et al., 2012), and orange juices (Pataro et al., 2011). 0.41c
These studies established the potential of the PL technique for achieving S7 59.0 6.0 35.0 7.84 ± 3.02 ± 0.03e
the pasteurization of fruit juices while affecting the phytochemicals 0.19e
S8 52.3 7.7 40.0 6.14 ± 2.73 ± 0.05abc
minimally within the matrix. 0.13ab
The present study desires to formulate a mixed fruit beverage using S9 55.4 8.4 36.2 6.34 ± 2.84 ± 0.07d
three mixture components: apple ber, carambola, and black table grape 0.16b
juices. The formulation was optimized, employing mixture design and S10 51.9 11.0 37.1 6.04 ± 2.84 ± 0.02d
0.18b
sensory acceptance as the response. Further, the efficacy of pulsed light
S11 48.1 11.9 40.0 6.41 ± 2.76 ± 0.01bc
(PL) and similar thermal treatments on various attributes of the mixed 0.23b
beverage was evaluated. The quality attributes tested were microbial S12 50.0 15.0 35.0 7.02 ± 2.72 ± 0.02a
traits, spoilage enzymes, color profile, vitamin C, total phenolic content, 0.29cd
and the antioxidant capacity of the mixed fruit beverage. S13 45.0 15.6 39.4 6.68 ± 2.70 ± 0.03a
0.24d
S14 45.0 15.6 39.4 6.78 ± 2.66 ± 0.05a
2. Materials and methods 0.19d
S15 46.6 18.4 35.0 5.90 ± 2.84 ± 0.01d
2.1. Raw materials and chemicals 0.33a
S16 46.6 18.4 35.0 5.78 ± 2.79 ± 0.02b
0.27a
Mature and fresh fruits (12–13◦ Brix) such as apple ber (Zizyphus
mauritiana), black table grapes (Vitis vinifera, Sharad seedless variety), # Crisp overall acceptability (COA) was measured through defuzzification of
carambola (Averrhoa carambola) were purchased (each 5 kg) from the sensory data provided by 16 semi-trained panelists in 5-point scale. Overall
hedonic score (OSH) was given by 60 consumers in 9-point hedonic scale.
local supplier near Matunga, Mumbai, India. All the fruits were free from
Different superscript letters within one column indicate significant (p ≤ 0.05)
any kind of physical or insect damage. The fruits were stored at 10 ◦ C for
differences among means determined by Tukey’s test.
a maximum of 4 days. The chemicals used in the experiment were of
analytical grade, and the microbial media were of bacteriological grade.
The chemicals and microbial media were purchased from HiMedia 2.3. Sensory analysis
Laboratories Pvt. Ltd., India.
The sensory analysis of the formulated beverages was conducted in
two parts. The sixteen formulations from the D-optimal mixture design
2.2. Preparation of mixed fruit beverage (MFB)
were placed in front of 60 untrained (27 male and 33 females; 23–55
years) and 16 semi-trained people (9 male and 7 females; 22–50 years)
The fruits were first manually deseeded and crushed through a home
for sensory evaluation. The protocol for sensory analysis was followed as
grinder (Model HL1645, Philips, India) at 6000 rpm for 5 min. The fruit
per Bureau of Indian Standards IS 6273: Guide for sensory evaluation of
pulp was further filtered using a stainless-steel screen 110 press-filter
foods, which does not mandate ethical approval (IS-6273, 1971). The
(Microfilt India, Mumbai, India) with a mesh size of 100 μm. The
evaluation performed by untrained people was termed a consumer test,
apple ber, carambola, and table grape juices’ average yield were 41, 68,
whereas the other was referred to as a panel test. The panelists were
and 60% w/w, respectively. The D-optimal mixture design was applied
selected from the staff and scholars at ICT Mumbai, India, who showed
for developing the mixed beverage (Jayachandran et al., 2015a, 2015b;
enthusiasm for sensory analysis. The samples were presented randomly
Bhalerao et al., 2020). The three components were: apple ber juice (X1,
in 100 mL flasks and were given scores by the assessors. In brief, the
45–59% v/v), carambola juice (X2, 1–20% v/v), black table grape juice
people for the panel test were screened out through the triangle test.
(X3, 35–40% v/v) (Table 1). The lower limits were selected to achieve a
After training for two weeks, the evaluation of those panelists became
minimum soluble solid content of 10 ◦ Bx, the minimum total soluble
reproducible. The method adapted for the sensory evaluation has been
solids prescribed by the Food Safety and Standards Authority of India
detailed by Bhalerao et al. (2020). The sensory attributes the panelists
(FSSAI, 2011). The upper limits for each were set concerning the pre­
decided to analyze were taste, appearance, aroma, after-taste, and
liminary sensory analysis. For instance, the presence of carambola
consistency of the beverage. The definitions of these attributes were
beyond 20% led to a sour taste that most of the panelists did not accept.
evolved from standards set by the International Organization for Stan­
Setting this 20% as the upper limit of carambola, the limits for other
dardization (ISO) (2008). Initially, from a set of sensory attributes, the
fruits were adjusted considering a maximum utilization of apple ber (an
top five critical attributes were screened out per the panelists’ agree­
under-explored fruit) and a TSS >10 oBx. As discussed further, the
ment. A 100-mL sample was served for sensory analysis, and the pan­
response assigned for modeling was the overall acceptance from the
elists provided their preference for each attribute of a sample on a
sensory analysis in the hedonic scale.

2
S. Chakraborty et al.
5-point categorized scale. Here, the categories referred to were ‘poor,’
‘fair,’ ‘good,’ ‘very good,’ and ‘excellent,’ respectively. They also
mentioned the importance of each sensory attribute while evaluating the
beverage. The categorizations were ‘not at all important,’ ‘somewhat
important,’ and ‘important,’ ‘very important,’ and ‘extremely important,
’ respectively. Fuzzy logic was applied on these sensory datasets from
the panelists using the methods detailed in Shrivastava and Chakraborty
(2018). For each formulation, the outcome from Fuzzy logic was a set of
similarity values (Sm) in the Fuzzy scale. The final categories for Fuzzy
scales were: ‘not satisfactory, ‘satisfactory,’ and ‘good,’ ‘very good, and
‘excellent’; the respective weightage (wF) assigned to each category was
1, 2, 3, 4, and 5. Further, a single parameter, called the crisp overall
acceptability (COA), was calculated using a method of weighted aver­
aging (Ross, 2005)
∑ borty et al., 2020; Vollmer et al., 2020). The fluence rate was varied at
S m × wF
COA = i ∑ (1)
Sm
i
The consumer acceptability or rating was conducted through a he­
donic scale. The consumers were asked to provide a score (SH) for each
of the sensory attributes (taste, appearance, aroma, after-taste, and
consistency) on a 9-point hedonic scale (1-dislike extremely, 9-like
extremely). Besides, the assessors were also asked to rate each sensory
attribute’s importance (IA) to judge the beverage. The scale for IA ranged
between 1 and 5 (1-not at all important, 5-extremely important).
Further, the overall hedonic scores (OSH) were calculated according to
Eq. (2) (Chakraborty et al., 2014).
∑ min (holding time). A thermocouple monitored the sample core tem­
SH × IA
OSH = i ∑ (2)
IA
i
The COA and OSH values for each formulation were considered as
two responses for the formulations evolved from the D-optimal mixture
design.
2.4. Response surface modeling
The response surface model was developed for both the responses as
a function of three constituents (x1, x2, and x3) of the blend. A special
cubic model (Eq. (3)) was the best fit model compared to others based on
the coefficient of determination (R2), adjusted R2, lack of fit, and min­
imum sum of squares (SSE) values.
COA ​ or ​ OSH = q1 x1 + q2 x2 + q3 x3 + q4 x1 x2 + q5 x1 x3 + q6 x2 x3 + q7 x1 x2 x3
(3)
q1 to q7 are the regression coefficients; x1, x2, and x3 are the L-pseudo
coded values of the apple ber, carambola, and table grape juice,
respectively.
2.5. Numerical optimization
The steepest ascent method was adopted in numerical optimization,
and it was iterated to obtain the maximum desirability value (Di) by
modifying X1 and X2 (Peinado et al., 2015). Being a mixture, fixing X1
and X2 automatically provides the value of X3. COA and OSH were taken
as responses to maximize the value of Di (Eq. (4)).
([ ]
COA − LCOA r1
[ ] )
OSH − LOSH r2 r +r
(4)
1
Di =
UCOA − LCOA
× ​
UOSH − LOSH
1 2
where L and U are the lower and upper limits of the individual responses
as denoted by the suffixes, r1 and r2 are the relative weightage of COA
and OSH, respectively, given in 5-scale (1-least weightage; 5-highest
weightage). The proportion of mixture components providing a
maximum value of Di was considered as the optimum blend. Further, the
optimized blends were analyzed to validate the predicted response
3
Food Bioscience 45 (2022) 101508
values.
2.6. Pulsed light treatment of MFB
The optimized blend was processed in batch mode pulsed light (PL)
equipment (X–1100 System, Xenon Corporation, Wilmington, MA, USA)
equipped with a linear xenon lamp (LH-840, type-B), as described by
Vollmer et al. (2020). A 50-mL beverage sample at 20 ◦ C was kept at 2.5
cm below the lamp on a glass cylinder, opened longitudinally, with a
sample layer thickness of 5 mm. A K-type thermocouple monitored the
surface temperature of the juice. A radiometer recorded the energy
irradiated (fluence rate) to the sample.
A two-factor-three-level full factorial design was applied for the PL
treatment performed at 2 kV with a pulse frequency of 3 Hz (Chakra­
18, 24, and 30 W cm− 2, and the samples were exposed for 33, 100, and
167 s (Table 4). The pulses applied corresponding to 33, 100, and 167 s
were 100, 300, and 500, respectively. The ON and OFF time was fixed at
144, 188, and 236 μs for the fluence rate of 18, 24, and 30 W cm− 2,
respectively. As a whole, there were 9 PL treatment combinations with
total fluence varied between 600 and 5000 J cm− 2.
2.7. Thermal treatment of MFB
The thermal treatment was given to a 50 mL sample packed into a
transparent LDPE pouch (80 μm thickness) dipped into a thermostatic
(T±0.5 ◦ C) water bath. The treatments were performed at 90 ◦ C for 1–5
perature. The come-up and cooling times were 127 s and 25 s, respec­
tively. The samples showing a 5-log reduction in resistant natural
microflora and complete inactivation of spoilage enzymes were reported
further.
2.8. Physicochemical characterization
The physicochemical profile of the beverage was assayed according
to the protocol described by Chakraborty et al. (2020). The pH was
assessed by a bench-top pH meter (Labserv, Fisher Scientific, India). The
total soluble solids (TSS, ◦ Brix) were measured using a handheld
refractometer (Erma 0–32 ◦ B, Japan). The CIE L*a*b* color profile was
recorded in reflection mode by a HunterLab colorimeter (LabScan XE,
Hunter Associates Laboratory, USA). The total color difference (ΔE*)
and browning index (BI) were estimated using Eqs. (5) and (6),
respectively.
√̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅
( * )2 ( )2
L1 − L*0 + (a*1 − a*0 ) + b*1 − b*0 (5)
2
ΔE =
[ ]
31 a* + 1.75 × L*
BI = (6)
0.172 5.645 × L* + a* − 3.012 × b*
The suffixes “1” and “0” in Eq. (5) stand for treated and unprocessed
(control) samples, respectively.
The phytochemical assays were conducted to estimate the total
phenolic content (TPC), antioxidant capacity (AOC), and concentration
of vitamin C or ascorbic acid (AA) in the sample. The spectrophoto­
metric method was employed using the protocol detailed by Chakra­
borty et al. (2015a) for all three assays. TPC was determined by taking
gallic acid as a reference compound and expressed as mg of gallic acid
equivalent (GAE). The antiradical scavenging potential (antioxidant
capacity) was quantified as mg of gallic acid equivalent antioxidant
capacity (GAEAC). L-ascorbic acid served as a standard compound for
vitamin C, and it was expressed as mg AA.
2.9. Microbial enumeration
The indigenous microbiota enumerated were aerobic mesophiles
S. Chakraborty et al. Food Bioscience 45 (2022) 101508

(AM), total coliforms (TC), and yeasts & molds (YM). The serial dilution-
pour plate enumeration method was adopted for this purpose (Chakra­
borty et al., 2015b). The AM and TC were incubated in plate count agar
(HIMEDIA® M091) and violet-red bile agar (HIMEDIA® M049),
respectively, for 24 h at 37 ◦ C. Yeast & malt agar (HIMEDIA® M424) was
used as a culture medium for YM, and the plates were incubated at 30 ◦ C
for 48 h. For the enumeration, 0.1 mL of the sample was plated out from
the serial dilution of 100 to 10− 6. The microbial count was expressed as a
colony-forming unit (CFU), and the detection limit was set as 10 CFU
mL− 1 samples.
2.10. Enzyme assay
The enzymatic assay was conducted for two oxidoreductases, viz.
polyphenol oxidase (PPO) and peroxidase (POD), following the protocol
discussed by Chakraborty et al. (2015b). The enzyme activity (A) was
expressed in %residual activity taking the enzyme activity in the un­
treated sample (A0) as reference.
2.11. Sensory analysis of PL and thermally treated beverages
A consumer sensory test using untrained assessors was conducted on
untreated, thermally treated, and PL-treated samples. The protocol for
the consumer sensory test has been briefed in section 2.3. The hedonic
scores (OSH) were used to understand the change in sensory attributes of
the beverage after thermal and PL treatments.
2.12. Statistical analysis
The treatments and analysis for all the samples were duplicated,
resulting in 4 replications for each. Analysis of variance (ANOVA) was
applied to visualize the significant difference between the mean values
at a 95% confidence interval. Tukey’s HSD test was employed for this
purpose. The principal component analysis (PCA) was carried out in
SPSS-v20 (IBM SPSS Statistics, USA).
0.91 (adjusted R2 = 0.86) and 0.85 (adjusted R2 = 0.74), respectively
(Table 2). Both the models showed an insignificant lack of fit (plof>0.09)
and a moderate F-value (>7.8). These data vouch for the special cubic
model as an adequately fit for both the groups of sensory data.
The model coefficients significantly influenced the COA and OSH
were linear components and the only interaction term between apple ber
and carambola juice (X1X2). However, the cubic interaction term (X1 ×
X2 × X3) was significant for the OSH model but not for COA. The positive
influence of the mixture components on the sensory properties was
evident from the coefficients in Table 2. The interaction between the
mixture components can also be visualized through the contours in
Fig. 1. This reflects that initially, the sensory score of the beverage
increased with an increase in either of the mixture components. How­
ever, after attaining an optimum, the sensory score (OSH) was
compromised due to suppressing the sweet taste of table grape juice by
the sour carambola component. It is reflected by the antagonistic
(negative) cubic term (X1 × X2 × X3) in the OSH model. Keeping the table
grape juice fixed, the interaction between apple ber and carambola
juices showed a synergistic (coefficient was 5.61) for OSH and antago­
nistic (coefficient was − 2.27) for COA. A similar trend between the
mixture components was reported for litchi-coconut water-lemon juice-
based mixed beverages (Jayachandran et al., 2015a, 2015b). The
antagonistic contribution of the cumulative mixture term was equally
supported in the case of pomegranate-muskmelon-amla-based mixed
fruit beverages (Bhalerao et al., 2020).
The numerical optimization utilized the maximum amount of
underexplored fruit juice such as apple ber juice. The relative weightage
for X1, X2, and X3 was decided as 4, 3, and 3, respectively (Table 3). The
optimized formulation was X1: X2: X3 = 59.0: 3.5: 37.5 with the
maximum desirability value of 0.898. The estimated COA and OSH
values were 3.2 and 7.6, respectively. Further, the optimized beverage’s
actual COA and OSH values were 3.23 and 7.55, respectively (Table 3).

3. Results & discussion

The crisp overall acceptability (COA) for the formulation ranged Table 2
between 2.66 and 3.29 (on a scale of 5), and it varied with the compo­ Estimated component coefficients for the special cubic model (Eq. (3)) and the
corresponding ANOVA data obtained from the D-optimal mixture design
sition of the beverage (Table 1). An increase in the apple ber juice
coupled with response surface methodology.
component in the formulation led to a higher COA value. For instance,
the beverage with X1: X2: X3 = 55.6: 4.4: 40.0 (sample S3) had the COA Term in Eq. 3 Model for overall hedonic Model for crisp overall
score (OSH) acceptability (COA)
value of 2.98 ± 0.02, whereas the blend with X1: X2: X3 = 48.1: 11.9:
40.0 (sample S11) scored the COA value of 2.76 ± 0.01. On the other Coefficient estimates (at p < Coefficient estimates (at p <
0.05) when mixture 0.05) when mixture
hand, the formulation scored a lower COA value when the carambola
components are in components are in
juice in the blend was more. Taking an example, the COA value for the
beverage with X1: X2: X3 = 46.6: 18.4: 35.0 (sample S16) was 2.79 ± L-Pseudo Actual L-Pseudo Actual form
form form form
0.02, whereas the blend with X1: X2: X3 = 59: 6: 35 (sample S6) had the
COA score of 3.06 ± 0.03. On a similar note, the consumers’ overall X1: Apple ber juice 6.88 − 0.47 3.68 0.067
X2: Carambola 5.18 − 21.55 2.94 1.309
hedonic scores (OSH) for the formulations ranged between 5.78 and
juice
7.84. The maximum OSH was obtained for the formulation with X1: X2: X3: Grape juice 20.97 0.18 1.22 − 0.0811
X3 = 59: 6: 35 (sample S7), whereas the lowest OSH value was estimated X1 × X2 5.61 0.50 − 2.27 − 0.0298
for the blend with X1: X2: X3 = 46.6: 18.4: 35.0 (sample S16). The trend X1 × X3 − 0.13** 0.01 0.61** 1.012 ×
followed by OSH was similar to the COA. To summarize, the OSH and 10− 3
X2 × X3 − 11.35** 0.59 0.93** − 0.0277
COA values, representative of consumer and panel preferences, indi­ X1 × X2 × X3 − 95.28 − 0.014 4.61** 6.722 ×
cated higher preferences for blends with minimal carambola juice and a 10− 4
maximum of apple ber juice. p-value (model) 0.0002 – 0.0132
F-value (model) 16.41 – 7.89
df (model) 6 6
3.1. Optimizing the formulation for MFB
p-value (lof) 0.1924 0.0941
df (lack of fit) 4 4
It was evidenced that the interaction between the individual com­ df (pure error) 5 5
ponents significantly influenced the sensory acceptance of the formu­ R2 (model) 0.91 – 0.85
lations. The responses viz. crisp overall acceptability (COA) and overall R2adj (model) 0.86 – 0.74

hedonic score (OSH) of the formulations were satisfactorily fitted to the df, degree of freedom; lof, lack of fit; **All model terms are not significant (p >
special cubic model (Eq. (3)). The R2 values for the OSH and COA were 0.05).

4
S. Chakraborty et al. Food Bioscience 45 (2022) 101508

Fig. 1. Contour plots showing the relative interaction among the three constituents of the mixed fruit beverage such as apple ber, carambola, and grape juices on two
different sensory scores; (a) overall hedonic score (OSH) obtained from 60 consumers, and (b) crisp overall acceptability (COA) from 16 semi-trained panelists.

Table 3
Summary of the numerical optimization criteria, optimized condition for obtaining the maximum crisp overall acceptability (COA) and overall hedonic score (OSH).
Variables Goal Lower limit Upper limit Rel Imp (ri) Optimization condition

Predicted Actual

Apple ber juice (mL) Maximize 45 59 4 59.0 59.0

Carambola juice (mL) In range 3.5 18.4 3 3.5 3.5
Grape juice (mL) In range 35 40 3 37.5 37.5
Crisp overall acceptability (COA) Maximize 2.65 3.28 5 3.20 3.23 ± 0.22
Overall hedonic score (OSH) Maximize 5.60 7.83 5 7.60 7.55 ± 0.24
Desirability (Di) Maximum 0 1 NA 0.898 NA

NA, not applicable.

3.2. Effect of pulsed light and thermal treatments on various attributes of
MFB
The intensities or fluence of the pulsed light treatment (PLT) con­
ditions varied between 600 and 5000 J cm− 2 (Table 4). At a fixed fluence
rate, the surface temperature was higher when the number of pulses was
more. The juice surface temperature varied from 30 ◦ C to 49 ◦ C when the
fluence varied from 600 J cm− 2 to 5000 J cm− 2, respectively. However,
the temperature within the sample was 21–32 ◦ C s (Table 4).
3.2.1. pH, acidity, and TSS of juice
The optimized formulation had the total soluble solids (TSS) of 12.0
± 0.2 ◦ Bx. The corresponding pH of the mixed fruit beverage was 3.90 ±
0.02 with a titratable acidity of 0.29 ± 0.01% citric acid (anhydrous).
The pH of the pulsed light-treated beverage ranged between 3.85 and
3.93, reflecting that the effect of pulsed light on the pH was not
statistically significant (p > 0.05). The pH of thermally treated samples
ranged between 3.89 and 3.92, which belonged to the same statistical
subgroup (p > 0.05). The titratable acidity also followed the same trend
as pH (statistically insignificant) (Table 5). The TSS range for PL and
thermally treated beverages were 11.7–12.2 ◦ Bx and 12.1–12.3 ◦ Bx,
respectively. Therefore, both the thermal and PL treatments did not
influence pH, TSS, and titratable acidity in the beverage. A similar trend
on acidity and TSS has been reported in the case of pineapple juice
(Vollmer et al., 2020), amla juice (Chakraborty et al., 2020), and
pineapple-amla-coconut water-based mixed fruit beverages (Dhar &
Chakraborty, 2020). The energy associated with the PL intensity
exposed to the juice was insufficient to induce ionic dissociation
releasing hydronium ions in the medium; thereby, the pH and acidity
remained unaffected. This might also be true for low dissociation
enthalpy of high molecular weight sugar into the monomeric unit. The
enthalpy associated with the thermal treatment of 90 ◦ C for 5 min was

Table 4
Summary of the pulsed light treatment details employed for the mixed fruit beverage.
Voltage Sample Treatment time Number of ON or OFF time Total Fluence (J/ Average Fluence rate (W/ Maximum temperature rise (ΔT,
(kV) (s)a pulses (μs) cm2)b cm2) ◦
C)c

2.0 PL1 33 100 146 600 18.00 ± 0.01 10.0 ± 0.1

PL2 100 300 146 1800 17.1 ± 0.1
PL3 167 500 146 2999 20.6 ± 0.2
2.0 PL4 33 100 188 801 24.00 ± 0.00 12.5 ± 0.2
PL5 100 300 188 2402 19.3 ± 0.2
PL6 167 500 188 4004 25.4 ± 0.1
2.0 PL7 33 100 236 1000 30.00 ± 0.01 16.8 ± 0.2
PL8 100 300 236 3000 22.7 ± 0.2
PL9 167 500 236 5000 29.0 ± 0.3
a
The pulse frequency was 3 Hz (3 pulses per second).
b
Total fluence = average fluence per pulse × number of pulses; Fluence rate = total fluence/treatment time.
c
This is the surface temperature of the beverage measured during the treatment and the sample temperature before the onset of pulsed light treatment was 20 ◦ C.

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S. Chakraborty et al. Food Bioscience 45 (2022) 101508

Table 5
Summary of the various physicochemical properties, color, and sensory attributes of the beverage after thermal and PL treatments.
Sample pH TSS (oBx) Titratable acidity (% citric acid) L* a* b* ΔE* BI OSH
a a a a g c d
UT 3.90 ± 0.02 12.0 ± 0.2 0.29 ± 0.01 21.84 ± 0.24 3.73 ± 0.13 6.68 ± 0.16 – 70.72 ± 0.9 7.5 ± 0.4d
TP1 3.92 ± 0.01a 12.0 ± 0.1a 0.28 ± 0.02a 25.83 ± 0.25f 4.91 ± 0.07h 6.83 ± 0.05d 4.16 ± 0.6b 69.39 ± 0.7cd 6.2 + 0.6b
TP2 3.89 ± 0.10a 12.1 ± 0.2a 0.29 ± 0.01a 25.87 ± 0.19f 4.85 ± 0.11h 7.34 ± 0.13f 4.23 ± 0.5b 70.15 ± 0.9d 5.8 ± 0.4a
PL1 3.91 ± 0.01a 12.0 ± 0.2a 0.29 ± 0.01a 22.19 ± 0.07b 3.45 ± 0.03f 6.65 ± 0.02c 0.8 ± 0.5a 70.11 ± 0.8d 7.5 ± 0.3d
PL2 3.90 ± 0.02a 12.0 ± 0.1a 0.28 ± 0.02a 24.48 ± 0.05e 1.32 ± 0.04d 6.73 ± 0.01cd 3.92 ± 0.5b 66.75 ± 0.8d 7.4 ± 0.2d
PL3 3.88 ± 0.02a 12.2 ± 0.1a 0.28 ± 0.01a 26.55 ± 0.02h − 0.31 ± 0.03b 7.08 ± 0.02e 6.56 ± 0.4d 64.86 ± 0.4a 7.0 ± 0.2c
PL4 3.86 ± 0.01a 12.1 ± 0.2a 0.29 ± 0.01a 23.34 ± 0.04d 3.26 ± 0.07e 6.38 ± 0.03b 0.84 ± 0.5a 68.64 ± 0.7c 7.3 ± 0.4d
PL5 3.88 ± 0.02a 12.2 ± 0.2a 0.28 ± 0.02a 26.04 ± 0.05f 1.38 ± 0.03d 5.97 ± 0.03a 4.02 ± 0.3b 64.89 ± 0.5a 7.3 ± 0.2d
PL6 3.87 ± 0.01a 12.2 ± 0.1a 0.28 ± 0.02a 27.49 ± 0.03g − 0.53 ± 0.02a 6.84 ± 0.04d 6.85 ± 0.5d 63.96 ± 0.9a 6.9 ± 0.3c
PL7 3.86 ± 0.01a 12.1 ± 0.1a 0.28 ± 0.00a 22.55 ± 0.02c 3.57 ± 0.06f 6.59 ± 0.02c 1.23 ± 0.4a 69.86 ± 0.7d 7.2 ± 0.4d
PL8 3.85 ± 0.02a 12.2 ± 0.2a 0.28 ± 0.01a 25.73 ± 0.03f 0.76 ± 0.02c 7.33 ± 0.03f 5.43 ± 0.4c 66.59 ± 0.9b 7.2 ± 0.3d
PL9 3.85 ± 0.01a 12.2 ± 0.0a 0.28 ± 0.02a 27.54 ± 0.03g − 0.52 ± 0.03a 8.21 ± 0.06g 7.02 ± 0.6d 65.98 ± 0.8ab 6.7 ± 0.4c

UT: Untreated beverage; TP1: Thermal treatment (90 ◦ C|4 min); TP2: Thermal treatment (90 ◦ C|5 min).
Different superscript letters within one column indicate significant (p ≤ 0.05) differences among means determined by Tukey’s test.

not adequate to induce any change in acidity and TSS of the beverage.
3.2.2. Microbial quality
The untreated beverage showed the aerobic mesophilic count (AMC)
and yeasts and mold count (YMC) of 5.75 and 5.15 log10 cfu mL− 1,
respectively (Fig. 2). The thermal treatment of 90 ◦ C for 4 min was
sufficient to reduce the YMC below the detection limit (DL of 10 cfu
mL− 1); AMC required 1 min more (90 ◦ C for 5 min) for the same. In the
case of pulsed light treated (PLT) samples, the samples treated with at
least 2400 J cm− 2 (24 W cm− 2 for 100 s) showed YMC below the
detection limit. Concomitantly, aerobic mesophiles showed higher
resistance, and a total fluence of 5000 J cm− 2 (30 W cm− 2 for 167 s) was
able to reduce the AMC below DL. The coliform count was below DL
even at the lower limit of the domain (600 J cm− 2). In this sense, the
beverages thermally processed at 90 ◦ C for 5 min (TP2 in Fig. 2) and PLT
at 30 W cm− 2 for 167 s (PL9 in Fig. 2) were considered microbially safe
samples. In the literature, Vollmer et al. (2020) reported a 5-log10
reduction in both YMC and AMC in pineapple juice while PL was treated
at 1479 J cm− 2. Chakraborty et al. (2020) recorded a similar extent of
inactivation for YM in amla juice at 1504 J cm− 2. On the other hand, for
a cocktail of fruit juices, Dhar and Chakraborty (2020) reported more
than 5-log reduction in both AMC and YMC after a PL treatment of 3143
J cm− 2. Similarly, Perscianowski (2021) also reported a 5.8-log reduc­
tion in E. coli P36 in kale juice at an absorbed fluence dose of 108.3 mJ
cm− 2.
The PL-induced lethality in the microbial count is demonstrated by
three similar effects: photo-chemical, photo-thermal, and photo-
physical effects (Ferrario et al., 2014). The UV portion (mainly UV-C)
is responsible for photo-chemical lethality within the microbial cell,
whereas the infra-red part contributes to the photo-thermal damage to
the cells. The modification in cytoplasmic membrane permeability
through shrinkage results from the photo-physical effect induced by
pulsation of high-energy light (Krishnamurthy et al., 2010). The
extended PL exposure (30 W cm− 2 for 167 s) to the sample led to a
temperature rise of 29 ◦ C, which is the contribution of the photo-thermal
effect. A higher resistance shown by aerobic mesophiles towards PL
exposure might be due to the variation in matrix pH, varying phenotype,
and genotype characteristics.

3.2.3. Enzyme activity

Fig. 2. Effect of various pulsed light and thermal treatments on aerobic mes­
ophiles (AM) and yeasts & molds (YM) population in the optimized mixed fruit
juice. The different alphabets above each bar denote that the corresponding
mean value differs significantly from the other mean value at a 95% confi­
dence interval.
The thermally treated (90 ◦ C for 4 min) beverage sample showed
10% PPO and 17% POD activity, whereas no enzyme activity was
detected in the sample after the treatment of 90 ◦ C/5 min (Fig. 3). Both
oxidoreductases (PPO and POD) in the beverage sample were sensitive
towards PL exposure. For instance, PPO inactivation was varied from 56
to 100%, and POD inactivation ranged between 67 and 100% within the
domain of 600–5000 J cm− 2. Obviously, at a specific fluence rate (say
30 W cm− 2), a greater extent of inactivation in both PPO and POD was
obtained for an increased number of pulses. At 24 W cm− 2, 62.4, 77.2,
and 87.9% PPO inactivation were reported when the sample was treated
at 801, 2402, and 4004 J cm− 2, respectively (Fig. 3). For the same sets of
treatment conditions, the extent of POD inactivation was 73.1, 83.1, and
99.2%, respectively. The sample treated at 30 W cm− 2 for 167 s (total
fluence of 5000 J cm− 2) was devoid of PPO and POD activity, and it can
be considered as the enzymatically stable sample. PPO and POD were
reported to offer resistance towards PL treatment to varying extents.

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S. Chakraborty et al. Food Bioscience 45 (2022) 101508

cm− 2 (24 W cm− 2 for 33 s). The PL treated samples exposed for a longer
treatment time (167 s) showed lower BI values but higher ΔE* values.
For a fixed fluence rate, the samples treated for a longer duration
showed higher lightness (L*) and lower redness (a*) values (Table 5).
This led to a modified ΔE* value for those samples. Cserhalmi et al.
(2006) categorized the color change in fruit products as well visible when
3.0 < ΔE* < 6.0. In this sense, the color change in the samples treated
within 1800–2402 J cm− 2 was well visible, whereas the samples exposed
at 3000–5000 J cm− 2 showed a greatly visible (6.0 < ΔE* < 12.0) color
change. Vollmer et al. (2020) recorded a minimal color change in
pineapple juice while exposed to 1472 J cm− 2 of PL treatment. Dhar and
Chakraborty (2020) reported a maximum ΔE* and ΔBI values of 6.4 and
5.6, respectively, in the mixed fruit beverage when exposed to 3142 J
cm− 2 of PL treatment. A significant change in a* values has been
observed in PL-treated coconut water (Donsingha & Assatarakul, 2018).
The color change in the pulsed light treated samples might result
from enzymatic browning reactions happening between polyphenols
and oxidoreductases. On the other hand, the nonenzymatic browning,
such as Maillard reaction products, could influence the browning index
and ΔE* in the thermally treated samples. The maximum temperature
rises in PL treated sample was 29 ◦ C, which may not be sufficient to
induce the heat-induced nonenzymatic browning in the sample (Chak­
raborty et al., 2020).

3.2.5. Antioxidant capacity and total phenolics

The untreated juice sample had a total phenolic content (TPC) of
722.7 mg GAE⋅L− 1 with an antioxidant capacity (AOC) of 622.4 mg

Fig. 3. Effect of various pulsed light and thermal treatments on polyphenol

oxidase (PPO) and peroxidase (POD) activity in the optimized mixed fruit juice.
The different alphabets above each bar denote that the corresponding mean
value differs significantly from the other mean value at a 95% confi­
dence interval.

Complete inactivation of PPO and POD was not achieved in pineapple

juice, even after treatment of 1479 J cm− 2 (Vollmer et al., 2020). In the
case of PL treated, amla juice showed no PPO and POD activity after
being treated at 3012 J cm− 2 (Chakraborty et al., 2020). However, a PL
exposure of 3143 J cm− 2 resulted in a 41 and 51% inactivation in PPO
and POD inside a fruit beverage cocktail (Dhar & Chakraborty, 2020).
The varying extent of inactivation of oxidoreductases in the sample
after thermal and PL treatments might be attributed to the mechanism of
denaturation in the enzyme. Thermal stress leads to the unfolding of
quaternary or tertiary structures and the loss of functional groups in the
enzyme (Fortea et al., 2009). On the other hand, enzyme inactivation by
PL exposure happens due to UV-induced photo-oxidation of protein,
leading to unfolding or aggregation of the functional group (Pellicer
et al., 2018; Wang et al., 2017).

3.2.4. Total color change (ΔE*) and browning index (BI)

The color profile of the untreated juice was L* = 21.84, a* = 3.73,
and b* = 6.68 with a browning index (BI) value of 70.72. The total color
change (ΔE*) value changed between 0.8 and 7.0 within the PL domain
of 600–5000 J cm− 2. For the thermal treatments of 90 ◦ C/4 min and
90 ◦ C/5 min, the ΔE* values were 4.16 and 4.23, respectively. The
respective BI values were 69.39 and 70.15, respectively (Table 5). The
PL treated samples exposed for 33 s showed the minimum ΔE* values. Fig. 4. Effect of various pulsed light and thermal treatments on total phenolic
However, the sample treated at 1000 J cm− 2 (30 W cm− 2 for 33 s) content (TPC) and antioxidant capacity (AOC) of the optimized mixed fruit
showed the higher ΔE* value (1.23) than the sample treated at 800 J juice. The different alphabets above each bar denote that the corresponding
values differ significantly at a 95% confidence interval of the mean.

7
S. Chakraborty et al.
GAEAC⋅L− 1. The thermal pasteurization at 90 ◦ C/3 min resulted in 31%
and 33% loss in TPC and AOC, respectively (Fig. 4). The enzymatically
stable thermally pasteurized juice (treated at 90 ◦ C/4 min) showed 36
and 39% losses in TPC and AOC, respectively. The TPC content in the PL-
treated juices was significantly decreased within 10–17%, whereas
retention of AOC was varied between 87 and 98%. Typically, a higher
loss in both TPC and AOC was reported at a fixed fluence rate in the
sample exposed to PL for a longer duration. The PL fluence of 5000 J
cm− 2 resulted in a maximum loss of 17% in TPC, whereas maximum
AOC loss (13%) in the juice was observed at 4004 J cm− 2 (24 W cm− 2 for
167 s) (Fig. 5). Given retention of phenolic compounds and antioxidant
capacity, PL treated samples were superior to the equivalent thermally
treated juice samples. A similar trend has been reported in amla juice
(Chakraborty et al., 2020), pineapple juice (Vollmer et al., 2020), mixed
fruit beverages (Dhar & Chakraborty, 2020). Retention in AOC in PL
treated (12–14 J cm− 2) apple samples was reported by Palgan et al.
(2011). About 93% correlation was there between TPC and AOC in the
samples. The reduction in the TPC and AOC in the PL-treated samples
might be due to UV portion-induced modifications in the phenolic
structures. The temperature rise due to the infra-red spectrum of the PL
and the photo-oxidation of phenolic compounds might show a
compromised AOC and TPC in the juice samples (Ghosh et al., 2014).
3.2.6. Vitamin C
The untreated beverage had a vitamin C content of 622.4 mg L− 1.
The thermal treatment at 90 ◦ C resulted in 41 and 47% loss in AA when
treated for 4 and 5 min, respectively (Fig. 5). The corresponding loss was
4–22.2% within the PL domain. A more significant AA degradation was
observed in the sample exposed to a higher fluence level. For instance,
the sample recorded 5.0, 8.8, and 13.1% loss in AA content when treated
at 24 W cm− 2 for 33, 100, and 167 s, respectively (Fig. 5). The maximum
AA loss of 22.2% was obtained for the PL-treated sample exposed to an
intensity of 5000 J cm− 2. The sample showed a compromised AA content
for a particular exposure time when treated at a higher fluence rate. For
example, for an exposure time of 100 s, the AA losses were 6.3, 8.8, and
15.4% when the fluence rate was 18, 24, and 30 W cm− 2, respectively.
PL-induced degradation of AA has been reported in the case of different
fruit juices. A maximum 29% loss in AA was obtained in pineapple juice
after being exposed at 1479 J cm− 2 (Vollmer et al., 2020). A similar
trend has been reported in the case of amla juice (Chakraborty et al.,
2020) and its mixed fruit beverage (Dhar & Chakraborty, 2020). The
negative influence of the UV-C component of the PL spectrum might be
the significant contributor to this degradation (Tran & Farid, 2004).
Besides, the photo-thermal effect, aerobic and metal-induced
Fig. 5. Effect of various pulsed light and thermal treatments on vitamin C
content of the optimized mixed fruit juice. The different alphabets above each
bar denote that the corresponding values differ significantly at a 95% confi­
dence interval of the mean.
8
Food Bioscience 45 (2022) 101508
photo-oxidation of AA might be the other reasons behind this trend
(Tikekar et al., 2011). The oxidases might also induce the degradation of
AA during the PL treatment, which eventually showed a compromised
AA content in the sample.
3.2.7. Sensory analysis
The OSH value of the untreated beverage was 7.5 ± 0.4. The values of
OSH for thermal and PL-treated beverages were in the range of 5.8–6.2
and 6.7 to 7.5, respectively (Table 5). The higher sensory acceptance of
PL treated beverage blends can be attributed to the higher L* values and
lower BI values. Interestingly, the OSH values of most of the PL-treated
beverages did not show any statistical difference from the untreated
beverage. In other words, the assessors placed the PL-treated beverages
on par with the untreated beverage. The pH, TSS, and titratable acidity
of the beverage also remained constant after the treatments, which was
also indicated in the consistency attributes of the beverages. To
conclude, the PL-treated beverages received higher hedonic scores than
the thermally treated beverages, which were also reflected in the
physicochemical attributes and color profile of the beverages.
3.2.8. Principal component analysis
The principal component analysis (PCA) suggested that two mutually
orthogonal principal components (PC1 and PC2) can describe more than
86.8% variability within the data set. The rotated biplot between the
PC1 and PC2 depicts that PC1 explains 62.3% variability, whereas PC2
recognizes 24.5% disparity within the entire dataset (Fig. 6). The ther­
mal sensitivity of phytochemicals is reflected by the position of un­
treated (UT) and thermally treated samples (TP1 and TP2) on the
opposite quadrant of PC1 (Fig. 6). Among the PL counterparts (<1000 J
cm− 2), PL1, PL4, and PL7 are clustered together, near the untreated
sample. This might be due to the minimal changes in biochemical at­
tributes and color profile compared to the raw samples (Michalak et al.,
2020). Among the responses, the AOC, AA, and TPC are clustered
together, depicting a similar sensitivity towards thermal and PL stress.
The correlation between these TPC, AA, and AOC data was more than
92%. A high correlation of 91% between ΔE* and L* was obtained, along
Fig. 6. Biplots between two principal orthogonal components (PC1 and PC2) in
a rotated space obtained from PCA of various quality attributes of the mixed
fruit juice during thermal and pulsed light (PL) treatments. The nomenclature
used for the treatments is summarized in Table 4. L*, lightness; a*, redness; b*,
yellowness; ΔE*, total color change; BI, browning Index; AA, ascorbic acid;
AOC, antioxidant activity; TPC, total phenolic content; PPO, polyphenol oxi­
dase activity; POD, peroxidase activity; AMC, aerobic mesophilic count; YMC,
yeast and mold count. (For interpretation of the references to color in this figure
legend, the reader is referred to the Web version of this article.)
S. Chakraborty et al.
with a 90% correlation between BI and a*. The modulation in lightness
values in the PL treated samples was majorly responsible for color
change in the beverage. However, a higher BI was observed for the
samples with more significant redness indices (a*). The data relating to
each microbial group and enzyme were highly correlated (>90%). This
indicates that a higher intensity of PL resulted in the inactivation of the
microbe and the enzyme. The top three commonality coefficients for
PC1 were 0.353, 0.351, and 0.345 for PPO, AMC, and YMC, respectively.
This vouches for the importance of enzymatic stability and microbial
safety of the product while comparing various quality attributes among
thermal and PL-treated samples.
4. Conclusions
The D-optimal mixture design coupled with panel and consumer
sensory data suggested the optimized blend as a mixture of apple ber:
carambola: black table grape juices = 59.0: 3.5: 37.5 v/v. The thermal
treatment (90 ◦ C/5 min) and pulsed light treatment of 167 s at 30 W
cm− 2 (5000 J cm− 2) resulted in complete inactivation of natural
microbiota (aerobic mesophiles, yeasts, and molds) and spoilage en­
zymes (polyphenol oxidase and peroxidase). The PL pasteurized (5000 J
cm− 2) sample retained 25, 27, and 19% more vitamin C, antioxidants,
and phenolic compounds, respectively, than the thermally pasteurized
(90 ◦ C/5 min) beverage. Although complete inactivation of microbes
and enzymes was possible by PL and thermal treatments, reports indi­
cate the possibility of growth of microbial spores or revival in enzyme
activity during storage. Therefore, the evaluation of the shelf-life of the
pulsed light treated beverage is of great importance.
Ethical approval
Ethics approval was not required for this research.
Data availability statement
Research data are not shared.
CRediT authorship contribution statement
Snehasis Chakraborty: Conceptualization, Supervision, Writing –
original draft, Revision - Reviewing and Editing, Funding acquisition.
Sagar Mahale: Methodology, Formal analysis, Investigation, Valida­
tion, Resources. Rishab Dhar: Methodology, Investigation, Writing –
review & editing, Visualization. Somnath Basak: Investigation, Writing
– review & editing, Revision - Reviewing and Editing.
Declaration of competing interest
The authors have declared no conflicts of interest for this article.
Acknowledgment
The project is funded by the Science & Engineering Research Board
(SERB), Department of Science and Technology, India, under the grant
ECR/2016/001414.
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