Journal of Food Engineering 224 (2018) 62e70

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Journal of Food Engineering

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

Kinetic modeling of ascorbic acid degradation of pineapple juice

subjected to combined pressure-thermal treatment
Santosh Dhakal a, V.M. Balasubramaniam a, b, *, Huseyin Ayvaz a, 1,
Luis E. Rodriguez-Saona a
a
Department of Food Science & Technology, The Ohio State University, Columbus, OH 43210, USA
b
Department of Food Agricultural and Biological Engineering, The Ohio State University, Columbus, OH 43210, USA

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

Article history: A study was conducted to investigate and model kinetic degradation of ascorbic acid in freshly prepared
Received 12 December 2016 pineapple juice subjected to various pressure (0.1, 300,450 and 600 MPa)-thermal (30, 75, 85 and 95
C)
Received in revised form treatment combinations. Experiments were conducted using a semi-custom made high pressure kinetic
26 November 2017
tester as well as an aluminum thermal kinetic tester. Thermal degradation of ascorbic acid was described
Accepted 19 December 2017
Available online 22 December 2017
with simple first order kinetics. The thermal rate constants (k 75 -95 ◦C, 0.1 MPa) and activation energy (Ea )
for ascorbic acid degradation reaction varied in the range of 0.004e0.006 per min and 14.22e29.78 kJ/
mol, respectively. Within the experimental conditions of the study (300e600 MPa at 30
C for holding
Keywords:
Ascorbic acid
times up to 15 min) high pressure processing did not alter ascorbic acid content (535.5e564.5 mg/kg).
High-pressure processing Combined pressure-thermal treatment (300e600 MPa at 75e95
C) degraded ascorbic acid with
Kinetics increasing thermal intensity and was modeled using first order fractional conversion kinetics model. The
½A
Thermal processing lower asymptote value ( ½A∞ ), rate constants (k 75 -95 ◦C, 600 MPa) and Ea were in the range of 77e85%, 0.108
0
Pressure-thermal treatment to 0.138 per min and 17.4e43.8 kJ/mol, respectively. Similarly, pressure sensitivity (DV s ) was ~0
Pineapple juice and 2.99 cm3/mol at 30 and 95
C, respectively. Knowledge gained from the study can be useful for food
processors to optimize high pressure treatment conditions for pineapple juice products.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction As high temperature induces degradation of ascorbic acid,

Modern health conscious consumers demand that, besides
ensuring microbiological safety, processed juices should also pro-
mote health, protect taste, flavor, bioactive compounds and nutri-
tional quality attributes (Tomas-Barberan and Gil, 2008). Ascorbic
acid is an essential nutrient as well as an antioxidant (Grosso et al.,
2013; Riccioni et al., 2012; Traber and Stevens, 2011). Unfortunately,
ascorbic acid present in freshly prepared juice is thermodynami-
cally unstable, and it undergoes deterioration during processing
and storage. In addition to loss of nutritive value, degraded prod-
ucts from ascorbic acid could form undesirable color in juice due to
browning reactions and thus deteriorate the quality (Bharate and
Bharate, 2014).
* Corresponding author. 333 Parker Food Science and Technology, 2015 Fyffe
Court, The Ohio State University, Columbus, OH 43210, USA.
E-mail address: balasubramaniam.1@osu.edu (V.M. Balasubramaniam).
1
Present address: Department of Food Engineering, Canakkale Onsekiz Mart
University, Canakkale 17020, Turkey.
increased interest has been focused on retention of ascorbic acid in
fruit juice and beverages with the application of alternative lethal
agents like high pressure. High pressure processing (HPP;
400e600 MPa at or below room temperature) is a recently
commercialized alternative food processing technology. It primar-
ily utilizes pressure (at ambient or chilled temperatures) as a lethal
agent to destroy vegetative forms of spoilage and pathogenic mi-
croorganisms during food pasteurization. HPP has been demon-
strated effectively in retaining variety of nutrients and bioactive
constituents in foods. In recent years, combined pressure-thermal
treatment has also been investigated for development of shelf-
stable low acid foods (pH > 4.5) or extended shelf life (ESL) foods
with retention of nutrients during processing. For acidic foods
(pH < 4.5), combined pressure-thermal treatment of modest in-
tensity (400e600 MPa at 45e90
C) is used to destroy acidophilic
spoilage bacteria or mold spores (Lee et al., 2002, 2006; Silva et al.,
2012; Balasubramaniam et al., 2016). During combined pressure-
thermal treatment, synergistic, antagonistic or additive effect
could occur on rate processes of food constituents, depending upon

https://doi.org/10.1016/j.jfoodeng.2017.12.016
0260-8774/© 2017 Elsevier Ltd. All rights reserved.
 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70 63

the pressure-temperature regime of these thermodynamic state
variables (Balasubramaniam et al., 2015; Gupta et al., 2011).
Kinetic models can be used as tools to describe quantitative
biochemical and nutritional changes in foods during processing.
Kinetic models can be linear or non-linear forms of rate law
equations. The rate process of food constituents is usually defined
by zero, first or second order kinetics (Heldman, 2013). In literature,
ascorbic acid degradation has been described by first order kinetics
(Dhuique-Mayer et al., 2007; Hiwilepo-van Hal et al., 2012; Nisha
et al., 2004; Polydera et al., 2003; Tola and Ramaswamy, 2015;
Vieira et al., 2016). Sometimes, a special case of first order kinetics,
known as fractional conversion model (Verbeyst et al., 2013) and
the nth order reaction model (Barba et al., 2012) have also been
reported.
Pineapple is one of the most popular non-citrus tropical fruits
that possess a wide array of nutritive and bioactive compounds
with antioxidants like ascorbic acid (USDA 2016 release 28). How-
ever, the systematic studies on combined pressure-thermal kinetics
of ascorbic acid degradation in pineapple juice are limited. There-
fore, the objective of this study was to evaluate the pressure effects
(with and without addition of heat) on ascorbic acid in pineapple
juice. In addition, kinetic parameters (rate constant, activation en-
ergy and activation volume) were estimated to develop a kinetic
model that could describe the changes in ascorbic acid at various
pressure (0.1e600 MPa)-temperature (30e95
C) regimes.
2. Materials and methods
(b) at constant temperature i.e 95
C at 300, 450 and 600 MPa for
0, 2, 5, 10 and 15 min.
Such pressure-thermal treatment combinations have been re-
ported to destroy acidophilic spoilage bacteria or mold spores
during the treatment of shelf-stable fruit products (Lee et al., 2002,
2006; Silva et al., 2012).
2.4. Thermal processing
Pineapple juice (6 mL) was filled in aluminum thermal death
time (TDT) tubes (inside diameter 15 - mm, 3 - mm thickness and
height 39 - mm) (Luechapattanaporn et al., 2004; Rajan et al.,
2006). TDT tubes were immersed in a constant temperature wa-
ter bath (Cole- Parmer) and processed at 75e95
C for 0e60 min
treatment times. Three replicates were used for each of the two
independent batches. A K-type thermocouple was inserted into one
of dummy TDT tube containing juice (control) for monitoring and
recording test sample temperature using a data logger thermom-
eter (HH 374, Omega Engineering Inc., Stamford, Connecticut, USA).
Come-up time for the thermal treatment was estimated as
1e1.25 min (75e95
C). Representative temperature history of
pineapple juice is presented in Fig. 1A.
After thermal treatment, TDT tubes were removed and imme-
diately cooled by immersing in ice bath. The samples were stored at
4
C up to 24 h prior to conducting ascorbic acid analysis.

2.1. Chemical and reagents

Certificed ACS grade acetic acid (99.9%), HPLC grade water,

Certified sodium hydroxide solution (1N) and hydrochloric acid
solution (1N) were purchased from Fisher Scientific (Pittsburg, PA, 90
USA). Ascorbic acid (99.5%), Reagent grade Sulphuric acid (96.3%), 75
Temperature(oC)

and Metaphosphoric acid (65%), HPLC grade water were purchased

from Sigma-Aldrich (St. Louis, MO, USA) and Phenolphathalein (1%) 60
indicator was purchased from LabChem Inc (Zelienople, PA, USA). 45
30
2.2. Pineapple juice and sample preparation
15
Whole pineapples (Ananas comosus) were purchased in bulk
0
from a local market in Columbus, Ohio (USA), peeled, cut in suitable 0 15 30 45 60
chunks, and juiced in a laboratory scale low speed (80 rpm) juicer
Time(min)
(VRT 400 HDS, Omega Products, Inc., Harrisburg PA, USA). The juice
was subsequently filtered with 170 mm sieve and stored in 200 mL
Low-density polyethylene (LDPE) pouches at 4
C up to a maximum
of 24 h until processing.

2.3. Experimental design 600 80

75
500
Temperature (oC)
Pressure (MPa)

Thermal processing of juice samples was conducted at 75, 85 70

400
and 95
C for treatment times of 0, 15, 30, 45 and 60 min. The 65
maximum temperature of 95
C was selected since pasteurization 300
60
process of shelf stable high acid fruit products (pH < 4.5) are typi- 200 55
cally conducted below 100
C (Silva and Gibbs, 2004). For high 100 50
pressure processing, pressure level of 300, 450 and 600 MPa at
30
C were selected for holding times of 0, 2, 5, 10 and 15 min. In 0 45
0 2.5 5 7.5 10 12.5 15 17.5
this study, the maximum pressure level of 600 MPa was selected as
Time (min)
it represents typical treatment condition for commercial high-
pressure pasteurization (Martínez-Monteagudo and Fig. 1. Representative pressure/temperature history during processing of pineapple
Balasubramaniam 2016). Likewise, experiments on combined juice.
pressure-thermal treatment were conducted as follows A. Thermal processing for 60 min ( / 75
C;  85
C; d 95
C; 0.1 MPa).
B. High pressure processing (600 MPa at 75
C) for 15 min ( d ) pressure, ( )/
temperature.
(a) under constant pressure i.e. 600 MPa at 75, 85 and 95
C for
0, 2, 5, 10 and 15 min, and
64 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70

2.5. High pressure processing as:

A high-pressure kinetic tester (PT-1 Avure Technologies Inc, 0 kT;P

Ao /Ao




!degraded products (1)
Kent, WA) was used for high-pressure treatment of pineapple juice.
Experiments were conducted using the procedure similar to those 0
Here, Ao is the ascorbic acid concentration in untreated (control)
described in our previous studies (Daryaei and Balasubramaniam, pineapple juice, Ao is its concentration at zero holding time. Then,
2013; Nguyen et al., 2014). Briefly, pineapple juice (2.75 mL) was the general rate law equation was used to evaluate the reaction
filled in several sterile flexible pouches. After removing the order as shown below (Heldman, 2013):
entrapped air bubbles, the packages were heat-sealed. Duplicate
pouches containing pineapple juice were then placed inside the d½A
insulated syringe (sample holder) and the void volume of the sy- rA ¼  ¼ kT;P ½An (2)
dt
ringe was filled with water. The syringe containing pouches were
then preheated or precooled to the specific temperature based on where, rA is the rate of ascorbic acid degradation, [A] is the ascorbic
the heat of compression values of the sample and target final acid concentration, kT;P is the rate constant at given temperature (T)
process temperature (Patazca et al., 2007; Rasanayagam et al., and pressure (P) and n (0, 1 and 2) is the order of reaction.
2003). Once the samples were loaded inside the pressure cham- The experimental data were normalized with ascorbic acid
ber, pressure-temperature history in the vicinity of sample within ½A
content at zero-minute holding time (½A t ). The integrated zero or-
the syringe was recorded. Representative pressure treatment his- 0

tory of pineapple juice is presented in Fig. 1B. The samples were der reaction could then be expressed as,
stored at 4
C.up to 24 h prior to ascorbic acid analysis.
½At 
¼ 1  kT;P ½A0 t (3)
2.6. Determination of pH, acidity and
Brix ½A0

where, ½At is the ascorbic acid concentration at time t and Ao is the

The pH was measured using a pH meter (Accumet XL 15, Fisher
initial concentration (zero holding time) of the reactant (ascorbic
Scientific, Mississauga, ON, Canada). Acidity of pineapple juice was
acid).
measured by titrating it with 0.1 N NaOH using phenolphthalein
Similarly, the integrated first - order reaction could be expressed
indicator (Helrich, 1990). oBrix value of the juice was measured by a
as,
refractometer (Reichert Analytical Instrument, Japan). Measure-
ments were taken at room temperature (z22
C). The total soluble   
½At ½A0 ¼ exp  kT;P t (4)
solids, pH and acidity of freshly prepared (control) pineapple juice
were estimated as 12.5 to 13 oBrix, 3.4 to 3.6 and 0.7e0.8% (w/w) as
and for the integrated second order reaction, it could be expressed
citric acid, respectively. These values were not also significantly
as,
different after various thermal or pressure-thermal treatment
conditions (data not shown).  1
½At ½A0 ¼ (5)
1 þ ½A0 kT;P t
2.7. Determination of ascorbic acid
In addition, a first order fractional conversion kinetics suggested
Ascorbic acid was determined by high-performance liquid by Verbeyst et al., (2013) was also evaluated:
chromatography (HPLC) as described by Landers (2014) and
 
Rodriguez et al. (1992). A volume of 4 mL of each juice sample was ½At ½A ½A  
mixed with 8 mL of an extraction solution (mixture of 3% meta- ¼ ∞ þ 1  ∞ exp  kT;P t (6)
½A0 ½A0 ½A0
phosphoric acid and 8% acetic acid). The resulting mixture was
centrifuged at 8000 g for 10 min at 4
C. The supernatant was where, ½A∞ is the non-zero equilibrium ascorbic acid concentration
collected and filtered through a 0.45 mm membrane filter into a at infinite process holding time.
2.5 mL amber glass vial.
Ascorbic acid levels were determined using a Schimadzu LC-10
(Kyoto Japan) system equipped with a SPD-M10AVP detector, a 2.8.1. Temperature sensitivity on rate constant
SIL-10A auto injector, an LC-10AS pump and degasser. Twenty mL of The temperature sensitivity of ascorbic acid degradation rate
each filtrate was injected into the system per each run. Isocratic was measured using Arrhenius model as suggested by Heldman
separation of ascorbic acid was achieved on a reverse phase C18 (2013):
column (3,5 mm, 4,6  150 mm2; Waters Corp., Milford, MA, USA).
Mobile phase was HPLC grade water acidified to pH 2.2 with sul-  
Ea
phuric acid with the flow rate was fixed at 1.0 mL/min. Elution of kT;P ¼ kA exp  (7)
RT
ascorbic acid was monitored at wavelength 245 nm. Identification
of the ascorbic acid was carried out by HPLC by comparing the where, kA  represent Arrhenius constant (frequency factor), Ea is
retention time and UVevisible absorption spectrum with those of the activation energy, R is the universal gas constant (8.3144 J/(mol
the standard solution prepared in metaphosphoric acid-acetic acid K)) and T is the temperature (K).
mixture using LC-10 software (Bellefonte, PA, USA). Calibration Equation (7) could also be reparametrized as;
curves were prepared at concentrations of 0, 50, 100, 150 and
" !#
200 mg/mL of ascorbic acid standard solution. This analysis was
0 Ea 1 1
done in duplicate for each sample. kT;P ¼ kref T;P exp   (8)
R T Tref
2.8. Kinetic models 0
where kref T;P
is the reaction rate constant at reference temperature,
Degradation of ascorbic acid in pineapple juice can be described Tref (363 K) and process pressure, P (0.1 or 600 MPa).
 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70 65

2.8.2. Pressure sensitivity on rate constant 1.1

The effect of pressure (P) on rate constant (kT;P ) at isothermal
conditions (T) was quantified by using Eyring-Polanyi equation 1
proposed by Martinez-Monteagudo and Saldan ~ a, 2014) as follows:
   0.9
vLn kT;P DV s
¼ (9)

[At]/[Ao]
vP T RT 0.8

where DVs is the activation volume. Using reference pressure, Pref 0.7
(500 MPa), Equation (9) could be reparametrized as;

 0.6
DV s -15 CUT
kT;P ¼ k"refT;P exp  P  Pref   (10) 0 15 30 45 60
RT 1 - 1.25 min Treatment time (min)

where, k"refT;P is the reaction rate constant at reference temperature

Fig. 2. Effect of thermal treatment on ascorbic acid in pineapple juice. Curves are fitted
(30 or 95
C) and reference pressure, Pref .
with first order kinetics. (⋯)75
C, (- - -) 85
C, (e) 95
C and (⋮) come up time.
Treatment time [process come-up time (CUT) and holding time].
2.8.3. Integrated kinetic models
The rate constant (kT;P ) presented in equations (8) and (10) were
combined into first order (equation (4)) and fractional conversion was lost during thermal (75e95
C) process come-up time. Subse-
model (equation (6)) to capture all the experimental data points. quently, the pineapple juice showed significant loss in ascorbic acid
For instance, the resulting first order (11 and 12) and first order content with increase in process temperature as well as treatment
fractional (13 and 14) equation were then expressed as; time. In comparison to unprocessed samples, samples treated at
75
C for 60 min lost 25% (w/w basis) ascorbic acid. When the
" !# !
 0 Ea 1 1 process temperature increased to 95
C, ascorbic acid loss was
½At ½A0 ¼ exp  kref T;P exp   t (11) increased up to 39% (w/w).
R T Tref
In literature, loss of ascorbic acid in pineapple juice/puree
  ranged between 25 and 60% depending upon the process temper-
 DVs
  ature and treatment time (Achinewhu and Hart, 1994; Chakraborty
½At ½A0 ¼ exp  k"refT;P :exp P  Pref t (12)
RT et al., 2015; Taoukis et al., 1998). Other factors that could also affect
the loss during thermal processing are sample preparation,
  " !# ! handling and transportation, as these operations could entrapped
½At ½A∞ ½A∞ 0 Ea 1 1
¼ þ 1 exp  kref T;P exp   t oxygen and enhance degradation at different rates (Uckiah et al.,
½A0 ½A0 ½A0 R T Tref 2009).
(13) Table 1A summarizes the degradation rate constants of ascorbic
acid in pineapple at different temperatures. We selected first order
reaction kinetics for which radj 2 and RMSE were ranged between

    0.88 to 0.92 and 0.0250 to 0.0391, respectively. The results showed

½At ½A∞ ½A DVs
 
that the first order rate constant (kT;P ; 0.0041 to 0.0060 per min)
¼ þ 1  ∞ exp  k"refT;P :exp P  Pref t
½A0 ½A0 ½A0 RT increased by 1.25 times with every 10
C rise in temperature from
(14) 75 to 95
C. Similarly, using the integrated Arrhenius model
0
(equation (11)), the rate constant (k90 C; 0:1 MPa ) was estimated to be
0.0057 per min. This values lies within the range reported by
various researcher for different fruit juices. For instance, the lowest
0
2.9. Data analysis rate constant (~k90 C; 0:1 MPa z 0.0025 per min) was reported for
amla and citrus juices by Nisha et al. (2004) and Dhuique-Mayer
0
The kinetic parameters were evaluated by non-linear regression et al. (2007). Highest rate constant (~k90 C; 0:1 MPa 0.036 per min)
method, without transforming the nonlinear equation into linear value was reported for orange juice (Vieira et al., 2016).
forms to obtain more precise estimate of the true population mean Arrhenius plot was also linear (r2 ¼ 0.99) within the investigated
of the original data as suggested by Boekel (1996). Equations were process temperature range (Fig. 3). Ea , the index of temperature
fitted by using Curve Fitting Tool Box, Matlab R 15b (Mathworks sensitivity of ascorbic acid degradation in pineapple juice at at-
Inc., Sherborne, MA, USA) and the regression analysis was per- mospheric pressure (0.1 MPa) was estimated in the range of
formed by Levenberg-Marquardt algorithms. 14e30 kJ/mol (Table 2). The lowest value of Ea was reported by
The statistical tools that were used to evaluate the adequacy of Nisha et al. (2004) for amla juice (17 kJ/mol). Ea required for
fit of the models were r2adj, RMSE, and significance (at 95% confi- ascorbic acid degradation in oranges and clementine were reported
dence level) of model parameters (½A∞ =½Ao ; kref ; Ea and DV s ) as 21 and 36 kJ/mol (Dhuique-Mayer et al., 2007; Vieira et al., 2016).
suggested by Boekel (1996). Models highest r2adj values and lowest Highest value (58 kJ/mol) was reported for guava pulp (Hiwilepo-
RMSE values were used as selection criteria. van Hal et al., 2012). Therefore, our results indicated that thermal
sensitivity of ascorbic acid in pineapple juice is similar to those of
3. Results and discussion oranges and clementine.

3.1. Thermal kinetics 3.2. High-pressure treatment effects at ambient temperature

The effect of thermal treatment on pineapple juice ascorbic acid The effect of pressure treatment (300e600 MPa at 30
C) on
content is shown in Fig. 2. A minor fraction of ascorbic acid (2e5%) ascorbic acid in pineapple juice is presented in Fig. 4. The ascorbic
66 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70

Table 1A
Comparison of reaction kinetic models and rate constantsa at different pressure-temperature combinations.

Rate process and Combined pressure- thermal treatment Thermal treatment

statistical
parameters

P (MPa) 600 450 300 0.1

T (oC) 75 85 95 95 95 95 85 75

kapp ¼ kT,P/[A]o 0.010 (0.009, 0.012 (0.01, 0.013 (0.012, 0.012 (0.011, 0.012 0.0052 (0.0049, 0.0044 (0.0040, 0.0039
(0th order) 0.011) 0.013) 0.015) 0.014) (0.010, 0.013) 0.0060) 0.0049) (0.0033,
0.0044)
r2adjusted 0.84 0.86 0.89 0.92 0.86 0.92 0.86 0.92
RMSE 0.0194 0.0219 0.0245 0.0167 0.0221 0.0362 0.0366 0.0245
kT,P 0.0107 (0.009, 0.013 (0.011, 0.015 (0.013, 0.014 (0.012, 0.013 (0.011, 0.0063 (0.0056, 0.0050 0.0042
(1st order) 0.012) 0.014) 0.017) 0.015) 0.014) 0.0069) (0.0044, 0.0056) (0.0038,
0.0046)
r2adjusted 0.86 0.88 0.90 0.93 0.87 0.91 0.88 0.92
RMSE 0.0182 0.0203 0.0228 0.0151 0.0212 0.0391 0.0347 0.0250
kapp ¼ kT,P. [A]o 0.011 (0.010, 0.014 (0.012, 0.016 (0.014, 0.015 (0.013, 0.014 (0.012, 0.0073 (0.0062, 0.0057 0.0046
(2nd order) 0.013) 0.016) 0.018) 0.017) 0.015) 0.0082) (0.0050, 0.0064) (0.0041,
0.0050)
r2adjusted 0.88 0.89 0.91 0.94 0.88 0.89 0.88 0.91
RMSE 0.0172 0.0189 0.0211 0.0136 0.0205 0.0428 0.0338 0.0260

kT,P is the ascorbic acid degradation rate constant, units per min for first order kinetics.
[A]o is the concentration of the ascorbic acid at zero minute holding time.
kapp is the apparent rate constant (units per min; normalized with [A]o to get the same units for all reaction orders).
r2adj is the adjusted coefficient of determination and RMSE is the root mean squared error.
a
Values in parenthesis represent 95% confidence interval.

Table 1B
Evaluation of first order fractional kinetic model and its parametersa at different pressure-temperature combinations.

Kinetic/statistical Combined pressure- thermal treatment Thermal treatment

parameters

P (MPa) 600 450 300 0.1

T (oC) 75 85 95 95 95 95 85 75
s s
kT,P (per min) 0.138 (0.061, 0.122 (0.047, 0.108 (0.029, 0.093 (0.026, 0.085 (0.003, 0.0063 0.014 0.0042s
0.216) 0.198) 0.187) 0.160) 0.167) (-0.01, 0.022) (-0.004, (-0.01,
0.032) 0.019)
[A∞]/[A]o 0.855 (0.816, 0.820 (0.760, 0.77 (0.687, 0.858) 0.783 (0.687, 0.78 (0.653, 0.91) 0.0000s (2, 0.56 0.0000s
0.890) 0.880) 0.879) 2) (0.14,0.97) (-3, 3)
r2adjusted 0.94 0.94 0.94 0.96 0.89 0.90 0.88 0.91
RMSE 0.0117 0.0139 0.0182 0.0107 0.0196 0.0402 0.0344 0.0256

kT,P is the ascorbic acid degradation rate constant at specific temperature and pressure.
[A∞]is the non-zero equilibrium ascorbic acid concentration at infinite treatment time.
[A]o is the concentration of the reactant (ascorbic acid) at zero minute holding time.
r2adj is the adjusted coefficient of determination.
RMSE is the root mean squared error.
s
Parameter estimate is not significant (p > .05).
a
Values in parenthesis represent 95% Confidence interval.

acid in HPP treated pineapple juice ranged from 540.5 to

566 mg kg1. Within the experimental conditions of the study,
1/T ( per Kelvin) there was no significant difference (p ¼ .098) in ascorbic acid con-
-5 tent between HPP (300e600 MPa at 30
C) samples and raw juice
0.00268 0.00271 0.00274 0.00277 0.0028 0.00283 0.00286 (control; 550 ± 14.5 mg kg1) irrespective of pressure holding times
-5.1 or pressure-come-up time. This indicated that pressure treatment
-5.2 at room temperature was not sufficient to cross the activation
barrier to initiate the ascorbic acid degradation reaction. In agree-
Ln(kTP)

-5.3
ment to our study, retention of ascorbic acid in HPP products like
-5.4 grapefruit juice, blood orange juice, mango pulp and bayberry juice
have been reported by various researchers (Kaushik et al., 2014;
-5.5
Torres et al., 2011; Uckoo et al., 2013; Yu et al., 2013).
-5.6 In contrast to our result, some previous studies showed that HPP
100 95 90 85 80 75 treated blueberry juice and fruit purees (strawberry, blackberry and
T (°C) pineapple) could reduce up to 8% ascorbic acid with increase in
pressure level from 200 to 600 MPa (Barba et al., 2013; Patras et al.,
Fig. 3. Arrhenius plot of Ln (kT,P) versus 1/T for heat treated pineapple juice. kT,P is the
2009; Chakraborty et al., 2015). Variation in pressure sensitivity
first order ascorbic acid degradation rate constant and T is the absolute temperature
(kelvin). Error bars represents 95% confidence interval. between juices was not unexpected due to several intrinsic and
 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70 67

Table 2
Estimation of Arrhenius and Eyring-Polanyi parametersa from integrated reaction kinetic models.

Model/Statistical Thermal Processing Combined Pressure thermal treatment

parameters (From integrated 1st order kinetics) (From integrated 1st order fractional kinetics)

Arrhenius model parameters Arrhenius model parameters

0
k90C;0:1MPa (per min) 0.0056 (0.0053, 0.0060) 0.116 (0.073, 0.160)
[A∞]/[A]o N/A 0.797 (0.76, 0.834)
Ea (kJmol1) 22.02 (14.2, 29.8) 30.6 (17.4, 43.8)
r2adjusted 0.90 0.94
RMSE 0.0332 0.0150
Eyring-Polanyi model parameters Eyring-Polanyi model parameters
k"95C;500MPa (per min) N/A 0.0961 (0.053, 0.139)
[A∞]/[A]o 0.773 (0.716, 0.829)
DVs(cm3mol1) 2.99 (5.26, 0.72)
r2adjusted 0.93
RMSE 0.0168

k"90C;0:1MPa is
ascorbic acid degradation rate constant at reference temperature (Tref ¼ 90
C).
[A∞]is the non-zero equilibrium ascorbic acid concentration at infinite treatment time.
[A]o is the concentration of the reactant (ascorbic acid) at zero minute holding time.
Ea is the activation energy.
r2adj is the adjusted coefficient of determination.
RMSE is the root mean squared error.
k"95C;500MPa is ascorbic acid degradation rate constant at reference pressure (Pref 500 MPa).
DVs is the activation volume.
N/A means not applicable.
a
Values in parenthesis represent 95% confidence interval.

57
Ascorbic acid (mg/100g juice)

56
1.1
55
54
1
53
[At]/[Ao]

52
0.9
51
50
-2.5 CUT 0 2.5 5 7.5 10 12.5 15 0.8
Control
0.4 - 0.8 min Treatment time (min)
0.7
-2.5CUT 0 2.5 5 7.5 10 12.5 15
Fig. 4. Effect of pressure treatment (HPP) at 30
C on ascorbic acid in pineapple juice. 0.8 min Treatment time (min)
(B)300 MPa, (,) 450 MPa, (D) 600 MPa, (◊) control (freshly prepared raw juice) and
(⋮) come up time. Error bars represents 95% confidence interval. No significance dif-
ference (p > .05) was observed between HPP samples and control at all pressure
holding time.

extrinsic factors such as composition, dissolved oxygen, pH, fruit 1.1

maturity and oxidation-reduction potential (Nagy, 1980). In this
study, we did not consider degradation of ascorbic acid in pressure- 1
thermal treated juice during extended shelf life. The degradation of
[At]/[Ao]

ascorbic acid in the product during extended storage conditions is

0.9
further influenced by the storage conditions (including storage
temperature, barrier properties of the packaging material) and this
merits future investigation. 0.8

0.7
3.3. Combined pressure-thermal kinetics -2.5CUT 0 2.5 5 7.5 10 12.5 15
0.4 - 0.8 min Treatment time (min)
The effect of combined pressure (600 MPa)-thermal (75e95
C)
treatment on ascorbic acid is presented in Fig. 5A. During pressure Fig. 5. Effect of combined pressure thermal treatment on ascorbic acid in pineapple
come-up time, similar to thermal processing, 2e5% loss of ascorbic juice. Curves are fitted with 1st order fractional conversion kinetics. Treatment time
acid was observed as process temperature increased from 75 to [(⋮) process come-up time (CUT) and holding time].
A. at constant pressure. (/ B) 600 MPa at 75
C, (- - - ,) 600 MPa at 85
C, (e D)
95
C at a pressure of 600 MPa. Increasing thermal intensity under
600 MPa at 95 oC.
pressure monotonically increased rate of ascorbic acid degradation. B. at constant temperature (⋯ B) 300 MPa at 95
C, (- - - ,) 450 MPa at 95
C, (e D)
The loss was rapid in early stages of pressure holding time, then 600 MPa at 95
C.
gradually slowed down, and became almost constant, possibly
reaching close to the equilibrium state. Consequently, the loss of
68 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70
ascorbic acid in pineapple juice at 75
C (for 15 min) was 13% (w/w)
when compared with untreated juice. Total ascorbic acid loss was
increased to 24% (w/w) at 95
C after 15 min holding time.
Our results showed that the ascorbic acid loss in pineapple juice
during combined pressure-thermal treatment were lower than
those reported in the literature. For instance, Taoukis et al. (1998)
reported up to 60% reduction of ascorbic acid in pineapple juice
processed at 75
C 600 MPa for 40 min. Chakraborty et al. (2015)
reported up to 24% reduction of ascorbic acid in pineapple puree
processed at 70
C-600 MPa for 20 min.
In addition to process parameters, other factors may contributed
for the variation in ascorbic acid loss within the same fruit (pine-
apple). It could be possible that ascorbic acid is less prone to
degradation in fruit juice than in more complex matrix like puree
owing to higher number of oxidizing enzymes and pro-oxidants in
the latter matrix. Next, the rate of pressurization could also influ-
ence the stability of ascorbic acid. Since, ascorbic acid degradation
occurred at faster rate during early phase of treatment process, the
slower pressurization rate might increase the degradation of
ascorbic acid. The higher losses of ascorbic acid reported by
Chakraborty et al. (2015) could also be due to slower pressurization
rate (5 MPa/s) as compared to our study (~11.12 MPa/s). Unfortu-
nately, the pressurization rate was not reported by Taoukis et al.
(1998). Next, the variation in selection of assay method can also
influence the result. In our study, ascorbic acid was determined by
HPLC method, whereas, Chakraborty et al. (2015) and Taoukis et al.
(1998) assayed ascorbic acid by colorimetric method using decol-
orizing reagent (2,6 dichlorophenol-indophenol dye), and by
enzymatic test kit, respectively.
3.4. Evaluation of reaction order and rate constant
In contrast to thermal kinetics, the experimental data of ascorbic
acid degradation with combined pressure thermal treatments can
be better described using first order fractional conversion model
(r2adjusted ¼ 0.89 to 0.96 and RMSE ¼ 0.0107 to 0.0196) rather than
simple first order kinetic model (Table 1B). In literature, this special
type of first order kinetics has also been reported in ozonized
strawberry juice (Tiwari et al., 2009) and other thermally treated
fruit and fruit products (Vieira et al., 2000; Vikram et al., 2005;
Verbeyst et al., 2013). In present study, the first order fractional
0
conversion rate constants (~k7595C;600MPa ) were in the range of
0.108e0.138 per min and independent of process temperatures
(Table 1B). Degradation of ascorbic acid during combined pressure-
thermal treatment may be influenced by other factors, such as
oxygen diffusion and its concentration in the sample (Verbeyst
et al., 2013).
½A
Likewise, the lower plateau concentration ( ½A∞ ) during com-
0
bined pressure thermal treatment was estimated in the range of
77e85%. These values were approximately equal to experimental
retention values at maximum holding time (15 min). Accordingly,
our results showed that combined pressure-thermal treatment
(600 MPa at 75e95
C) could be a better option when thermal
treatment (0.1 MPa at 75e95
C) needs considerably longer treat-
ment time (>40 min) to preserve the juice. For instance, treatment
time of ~62 min at 88
C is needed to achieve 5 log reduction of
ascospores of Neosartorya fischeri, a spoilage fungus in thermally
treated pineapple juice (Silva and Gibbs, 2004).
Therefore, in spite of higher rate constant, combined pressure-
thermal treatment could still preserve higher fraction of ascorbic
acid than thermal treatment. This type of specific retention char-
acteristics could not be explained from simple first order kinetics.
These results also illustrated that proper selection of kinetic model
is critical to understand ascorbic acid degradation process, which,
otherwise, would lead to erroneous interpretation.
3.5. Evaluation of temperature sensitivity
The Ea of ascorbic acid degradation in pineapple juice during
combined pressure-thermal treatment ranged from 17.4 to 43.8 kJ/
mol (Table 2). It is similar to that estimated during thermal pro-
cessing as described in Sub-section 3.1. This value was lower than
that reported by Taoukis et al. (1998) who conducted at lower
treatment intensity range (0.1e600 MPa at 40e75
C) in pineapple
juice (45e71 kJ/mol). Variation in Ea values is not an unexpected
result and has also been reported for other juices in the literature.
For instance, Johnson et al. (1995) reported Ea as high as 128 kJ/mol
for orange juice while Vieira et al. (2016) reported as low as 21 kJ/
mol for the same juice.
3.6. Evaluation of pressure sensitivity
With HPP (300e600 MPa at 30
C) treated pineapple juice, the
pressure sensitivity of ascorbic acid (DV s ~0) was considered
negligible as the corresponding rate constants were negligible (~0)
at all pressure levels. Fig. 5B shows the effect of change in pressure
on ascorbic acid at constant elevated temperature (95
C). In
contrast to HPP at 30
C, the degradation rate of ascorbic acid at
95
C showed a modest increase (DV s z-0.72 to 5.2 cm3/mol)
with increase in pressure level (Fig. 6). Our results are also com-
parable with DV s reported by Taoukis et al. (1998) in the range
of 3.26 to 6.14 cm3/mol in different juices (grape fruit, pine-
apple) and model foods. In addition, DV s having comparable
values ranging from 3.1 to 9.69 cm3/mol were also reported for
other antioxidants like anthocyanins in raspberry paste and blue-
berry juice (Verbeyst et al., 2011; Buckow et al., 2010). The negative
DV s could be due to oxidative reaction favored by combined
pressure-thermal treatment (GarcíaTorres et al., 2009).). It could be
possible that as the aerobic reaction was ceased due to depletion of
oxygen concentration in the medium, the oxidative reaction was
thermodynamically unfavorable and the ascorbic acid retention
value reached the asymptotic point. The effect of dissolved oxygen
concentration and temperature on ascorbic acid autooxidation has
been reported (Eison-perchonok and Downes, 1982). More studies
are necessary to understand the combined effect of dissolved ox-
ygen concentration, and pressure-thermal treatment on ascorbic
acid autooxidation.
4. Conclusion
Within the experimental conditions of this study, pressure
treated pineapple juice (300e600 MPa at 30
C) did not induce
-1.5
Ln(kT,P, per min)
-2.5
-3.5
250 300 350 400 450 500 550 600
Pressure (MPa )
Fig. 6. Eyring- Polanyi plot of Ln (kT,P) versus P for combined pressure-thermal treated
pineapple juice at 95
C. kT,P is the ascorbic acid degradation rate constant (first order
fractional) and P is the pressure (MPa). Error bars represents 95% confidence interval.
 S. Dhakal et al. / Journal of Food Engineering 224 (2018) 62e70
ascorbic acid degradation reaction and preserved ascorbic acid
irrespective of applied pressure level (up to 600 MPa) and holding
time (up to 15 min). Thermal treatment alone (at 0.1 MPa) degraded
(up to 39% loss) ascorbic acid as a function of temperature
(75e95
C) and treatment time (up to 60 min), following first order
kinetics. Combined pressure-thermal treatment increased the
ascorbic acid degradation rate and can be described using first or-
der fractional conversion model. Pressure sensitivity (DV s )was
 0 and -2.99. cm3/mol at 30 and 95
C, respectively. Activation
energy (Ea) for ascorbic acid degradation for both thermal and
combined pressure-thermal treatments were similar. More
research is needed to understand the influence of product attri-
butes (pH, sugars, salts, reactant (ascorbic) concentration, dissolved
oxygen, oxidative enzymes, amino acids and metal catalyst) on the
oxidative degradation of ascorbic acid during combined pressure-
thermal treatment. Such studies help the processors to formulate
pressure-thermal processed juices that retain maximum ascorbic
acid.
Acknowledgements
Research support to The Ohio State University Food Safety En-
gineering laboratory (u.osu.edu/foodsafetyeng/) was provided, in
part, by USDA National Institute for Food and Agriculture HATCH
project OHO01323 and the food industry. Authors are grateful to
the OSU Dr. Rodriguez-Saona's food analysis laboratory for access to
HPLC equipment. References to commercial products or trade
names are made with the understanding that no endorsement or
discrimination by The Ohio State University is implied.
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