Food Chemistry 128 (2011) 10871093

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Effect of microwave pretreatment on the kinetics of ascorbic acid degradation

and peroxidase inactivation in different parts of green asparagus
(Asparagus ofcinalis L.) during water blanching
Hong Zheng, Hongfei Lu
College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China

a r t i c l e

i n f o

Article history:
Received 6 November 2010
Received in revised form 17 February 2011
Accepted 30 March 2011
Available online 12 April 2011
Keywords:
Asparagus ofcinalis L.
Ascorbic acid
POD
Blanching
Microwave

a b s t r a c t
In this study, the effects of two blanching conditions on ascorbic acid (AA) and peroxidase (POD) in different segments of asparagus (bud, upper, middle, and butt) were investigated. The blanching treatments
were: blanching in water at 70, 80 and 90 C (WB); microwave heating (900 W, 30 s) followed by water
blanching (MW + WB). AA degradation and POD inactivation in all segments of asparagus for both treatments are well described by rst-order models. The degradation rate of AA and POD is gradually
increased from butt to bud segment of asparagus. In addition, MW pre-treatment could increase the Ea
of AA degradation and decrease the Ea of POD inactivation during water blanching of asparagus. Therefore, it is recommended that the different segments of asparagus should be subjected to different blanching times, and MW pre-treatment could be applied for alleviating AA degradation and accelerating POD
inactivation during blanching, cooking and pasteurisation in water.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
Asparagus (Asparagus ofcinalis L.) is a green vegetable with
high antioxidant activity compared with other commonly consumed vegetables (Vinson, Hao, Su, & Zubik, 1998). However, this
vegetable is highly perishable due to its high respiratory activity
that continues after harvesting. Therefore, it is often processed to
reduce post-harvest losses and increase the economic return for
farmers. Blanching is one of the many processes that take place
during the preparation of untreated vegetables for preservation
processes such as canning and freezing. Hot-water blanching is
by far the most popular and commercially adopted process as it
is the simplest and most economical technique. Its primary purpose is to inactivate enzymes and destroy vegetative microbial
cells, allowing stabilisation and product quality retention during
storage (Canet, 1989). Despite its preservative benets, it may also
have some negative effects on product quality, such as excessive
loss of texture, undesirable changes in colour, nutritional losses
and degradation of antioxidant properties (Arroqui, Rumsey, Lopez,
& Virseda, 2001; Oboh, 2005; Selman, 1994).
Peroxidase (POD) catalyses the oxidation reactions using either
peroxides or oxygen as the hydrogen acceptor (Hemeda & Klein,
1991). It is recognised as being one of the most heat-stable
enzymes and is widely used as an index of blanching (Baheci,
Serpen, Gkmen, & Acar, 2005). If this enzyme is inactivated, other
Corresponding author. Tel.: +86 0579 8228 2284.
E-mail address: luhongfei63@yahoo.com.cn (H. Lu).
0308-8146/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.03.130

enzymes responsible for tissue degradation will also be inactivated

(Koksal & Glin, 2008; Soysal & Sylemez, 2005). In addition,
ascorbic acid (AA) is an important component of our nutrition
and reportedly reduces the risk of arteriosclerosis, cardiovascular
diseases and some forms of cancer (Harris, 1996) because of its
antioxidant capacity (Glin, Bursal, Sehitoglu, Bilsel, & Gren,
2010). However, AA is one of the most labile vitamins. It is easily
oxidised by air, heat, oxidising enzymes, and multivalent cations
(Arroqui, Rumsey, Lopez, & Virseda, 2002; Gkmen & Serpen,
2007). It is generally observed that if AA is well preserved, the other
nutrients are also well retained (Lin, Durance, & Scaman, 1998).
Therefore, this research used POD and AA as the indicator of
enzymes and nutrients to evaluate asparagus quality during water
blanching after microwave pre-treatment, respectively. The objectives of the present work were (i) to evaluate the kinetics of AA degradation and POD inactivation in different parts of green asparagus
during two blanching treatments, namely, water blanching and
microwave heating followed by water blanching, and (ii) to study
the effect of microwave pre-treatment in alleviating AA loss and
accelerating POD inactivation during water blanching of asparagus.

2. Materials and methods

2.1. Sample preparation and blanching process
Fresh asparagus (A. ofcinalis L. var. UC800) was harvested from
a local farm in Jinhua (Zhejiang, PR China) and transported under

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H. Zheng, H. Lu / Food Chemistry 128 (2011) 10871093

refrigeration at 8 C for 30 min to the laboratory. Spears of uniform

diameter (0.5 0.1 cm) at the base and length (20 cm) were used in
this experiment. The spears, after being sorted for size and length,
were washed with tap water and drained. All spears were divided
into two equal groups subjected to microwave (30 s) and
non-microwave heating. In the microwave treatment, every time
approximately 200 g of asparagus was heated in the microwave
oven (Galanz WD900ASL23-5S, 23L, 900 W). After cooling to room
temperature, the spears were separately placed in two distilled
water baths set at 70, 80 and 90 C. The spears were separately
removed at different blanching times and immediately cooled in
running water at 5 C. Prior to measurement of ascorbic acid and
POD activity, the spears were cut into four segments (bud, upper,
middle, and butt segments) at room temperature as described in
our previous study (Lu et al., 2010).
2.2. Chemicals

2006; Frias & Oliveira, 2001; Giannakourou & Taoukis, 2003; Uddin, Hawlader, Ding, & Mujumdar, 2002; Zheng & Lu, 2011) for
evaluating AA degradation during food processing and storage.
Therefore, the kinetic data of POD activity inactivation and AA
degradation were analysed with rst-order kinetics using the following equations:

C t =C 0 expk  t

where Ct and C0 are the POD activity or AA content at time t and

zero, respectively; k is the rst-order rate constant; and t is the
blanching time (s).
The Arrhenius equation is the most widely accepted method of
accounting for the temperature dependence of the rate constant in
food systems. The temperature and the rate constant k are related
according to the Arrhenius equation:

K A expEa =RT

Ascorbic acid (purity P99%), 2,6-dichlorophenol indophenol

(purity P99%), guaiacol (purity P98%), H2O2 solution (purity
P30%) and bovine serum albumin (purity P99%) were purchased
from Shanghai Sangon Biological Engineering Technology & Services Co., Ltd. (Shanghai, China).

where Ea is the activation energy of the reaction; R is the gas

constant; T is the absolute temperature; and A is the pre-exponential constant.
The decimal reduction time (D-value) is dened as the treatment time needed for 90% degradation of its original value. The
D-value was calculated as follows:

2.3. Determination of ascorbic acid (AA)

The AA content in the spear segments was determined based

upon the quantitative discolouration of 2,6-dichlorophenol indophenol titrimetric method as described in AOAC methodology
No. 967.21 (AOAC 2000, chap. 45). The AA content was measured
in triplicate and expressed as mg/100 g fresh weight (FW).

where k is the rst-order rate constant.

2.4. Preparation of enzyme extracts

Crude extract preparation was carried out according to the
method described by Gonalves, Pinheiro, Abreu, Brando, and Silva
(2007). Raw and blanched asparagus samples (5 g each) were
weighed into 45 ml of 1 M sodium chloride solution. The samples
were ground gently in a mortar surrounded by chipped ice. The
homogenate was centrifuged in polypropylene tubes at 3500 rpm,
using a cold centrifuge (Hettich, Tottlingen, Germany), at 4 C for
10 min. The supernatants were then ltered using 1.2 lm
membrane lters (Whatman) and kept on ice until the analysis.
2.5. Determination of peroxidase (POD) activity and protein content
PPO activity was assessed in triplicate based on a modied
method of Gkmen, Baheci, Serpen, and Acar (2005). The substrate solution was a mixture of 0.1 ml of guaiacol and 0.1 ml of
H2O2, made up to 100 ml with 0.1 M acetate buffer (pH 5.50).
The substrate solution (4.9 ml) was transferred into a cuvette and
the reaction was started by adding 0.1 ml of crude enzyme extract.
One unit of POD activity was dened as an increase in absorbance
of 0.001 at 420 nm per minute per mg of protein under the assay
conditions at 37 C (Sisecioglu et al., 2010).
Protein content of the enzyme extracts was determined according to Bradford (1976) using bovine serum albumin as a standard
(Glin, Kfrevioglu, & Oktay, 2005)
2.6. Kinetic models of POD activity inactivation and AA degradation
For the inactivation of POD activity, Morales-Blancas, Chandia,
and Cisneros-Zevallos (2002) reported rst-order inactivation
kinetics showed a good t during thermal treatments of green
asparagus. In addition, the rst-order kinetic model has been also
applied by numerous researchers (Burdurlu, Koca, & Karadeniz,

2:303
k

2.7. Statistical analysis

Data were expressed as the mean standard deviation of three
measurements. Differences between means were analysed with
Duncans test using the SAS software package (version 8.1). A signicant difference was considered at the level of p < 0.05.
3. Results and discussion
3.1. Kinetic models to describe changes in AA and POD activity of
different parts of asparagus
First-order kinetic models were used to treat the resulting data
of AA degradation in different parts of asparagus during WB and
MW + WB as represented in Fig. 1. These models seemed to be suitable to describe AA degradation in this study because of the high
determination coefcients (R2 = 0.81630.9426). In fact, many
researchers reported that AA degradation of food materials are well
described by rst-order models (Frias & Oliveira, 2001; Giannakourou & Taoukis, 2003; Zheng & Lu, 2011). Fig. 2 shows that the
degradation of POD activity in all segments of asparagus also
followed rst-order reaction kinetics (R2 = 0.89680.9942). For
other vegetables such as carrots, potatoes, tomato and watercress,
a rst-order model was also used to describe the enzyme inactivation (Anthon & Barrett, 2002; Anthon, Sekine, Watanabe, & Barrett,
2002; Cruz, Vieira, & Silva, 2006).
Fig. 3 illustrates contour plots of rate constants (k) of AA
degradation and POD inactivation as a function of blanching temperature and different parts of asparagus for both treatments. As
can be seen from Fig. 3A and B, the k of AA loss increases with increased blanching temperature, but it decreases gradually from
bud to butt segment of asparagus. This result suggests that bud
segment is more liable to lose AA than other segments, which in
accordance with our previous ndings in green asparagus during
thermal treatments (Zheng et al., 2011). The loss in AA content
during blanching could be attributed to the fact that AA is very soluble in water and not stable at high temperature (Nagy & Smooth,
1977). Therefore, one possible explanation for these results is that

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Fig. 1. Residual ascorbic acid changes in bud (A), upper (B), middle (C), and butt (D) segments of asparagus during water blanching (WB) and microwave heating (900 W, 30 s)
followed by water blanching (MW + WB) at 70 C (j), 80 C (d), and 90 C (N).

the bud segment of asparagus provides a poorer texture than other

segments, which increases the penetration of AA and decreases
heat resistance. Torreggiani and Bertolo (2001) revealed that fruit
texture improvement could improve pigment, colour, vitamin,
and aroma retention during air dehydration and frozen storage.
In addition, Lau, Tang, and Swanson (2000) observed that the reaction rate constants of the thermal degradation of asparagus texture

gradually increased from the base to the tip of the asparagus

spears. According to Fig. 3C and D, the results are similar to those
obtained on the degradation of AA, that is, the k of POD inactivation
increased with blanching temperature and decreased from bud to
butt segment of asparagus. Therefore, it is recommended that the
bud segment of asparagus should be subjected to a relatively short
period of time during blanching, cooking and pasteurisation for

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H. Zheng, H. Lu / Food Chemistry 128 (2011) 10871093

Fig. 2. Residual POD activity changes in bud (A), upper (B), middle (C), and butt (D) segments of asparagus during water blanching (WB) and microwave heating (900 W, 30 s)
followed by water blanching (MW + WB) at 70 C (j), 80 C (d), and 90 C (N).

preserving AA content and inactivating POD activity. Moreover, the

different segments of asparagus should be blanched with different
times for preserving its quality.
Activation energies (Ea) were calculated by using Arrhenius
plots of POD inactivation and AA degradation in asparagus given
in Fig. 4 and found higher in the middle and butt segments than
that of the bud and upper segments (Table 1). A similar result

was obtained in our earlier study (Zheng et al., 2011) for AA loss
during thermal treatment of asparagus. High Ea in the middle
and butt segments indicated that POD inactivation and AA degradation was more temperature dependent than the other segments.
A possible reason for this is to be found in the texture of different
segments of asparagus, that is, a better texture of the middle and
butt segments improve resistance to heat. Hayakawa and Timbers

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1091

Fig. 3. Contour plots showing the effect of blanching temperature and different parts of asparagus on rate constant of ascorbic acid degradation (A and B) and POD
inactivation (C and D) during water blanching (WB) and microwave heating (900 W, 30 s) followed by water blanching (MW + WB).

Fig. 4. Arrhenius plots of ascorbic acid degradation (A and B) and POD inactivation (C and D) in different segments of asparagus (bud, upper, middle and butt) during water
blanching (WB) and microwave heating (900 W, 30 s) followed by water blanching (MW + WB).

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H. Zheng, H. Lu / Food Chemistry 128 (2011) 10871093

Table 1
Activation energies (Ea) of ascorbic acid (AA) degradation and peroxidase (POD)
inactivation in bud, upper, middle, and butt segments of asparagus during water
blanching (WB) and microwave heating (900 W, 30 s) followed by water blanching
(MW + WB).
Ea (kJ mol1)

AA
POD

WB
MW + WB
WB
MW + WB

Bud

Upper

Middle

Butt

18.44ba
23.12a
15.52a
15.23a

19.14b
22.70a
20.66a
20.59a

21.07a
22.68a
23.86a
18.51b

20.81a
21.29a
22.83a
20.61b

a
Different lower case letters indicate signicant differences (P < 0.05) between
WB and MW + WB.

Fig. 6. The percentage retention of ascorbic acid (AA) in bud, upper, middle, and
butt segments of asparagus during MW + WB (microwave heating followed by
water blanching) and WB (water blanching) at 70 C (A), 80 C (B), and 90 C (C) for
inactivating 90% of POD. Different lower case letters indicate signicant differences
(P < 0.05) between WB and MW + WB.

Ea of POD inactivation was decreased during MW + WB, but there

were no signicant differences for the bud and upper segments.
The reason for these results will need further study before they
can be satisfactorily accounted for.
3.2. The effect of blanching method on the degradation of AA and POD
activity in different parts of asparagus
Fig. 5. The D-value of POD inactivation in bud, upper, middle, and butt segments of
asparagus for both treatments (WB: water blanching; MW + WB: microwave
heating followed by water blanching) at 70 C (A), 80 C (B), and 90 C (C). Different
lower case letters indicate signicant differences (P < 0.05) between WB and
MW + WB.

(1977) and Lau et al. (2000) reported that the hardness of asparagus gradually decreased from the base to the tip of asparagus.
According to Table 1, the Ea of AA degradation during water
blanching was increased for all segments of asparagus after microwave pre-treatment, although a non-signicant difference was
observed for the middle and butt segments. On the contrary, the

POD is the most thermally resistant enzyme in vegetables, and

for this reason, complete inactivation of POD requires considerably
long time, resulting in heavy loss of nutrients. Therefore, Singh and
Chen (1980) reported that 90% inactivation of POD as sufcient to
prevent any deterioration in food and recommended the same as
the optimum level for blanching. Fig. 5 shows the D-value of POD
inactivation in different segments of asparagus for both
treatments. These results suggest that signicant decreases
(P < 0.05) in the D-value of POD in all segments of asparagus are
observed during MW + WB at all temperatures used in this study.
This phenomenon results from a decrease in the Ea of POD inactivation during MW + WB, although this is the subject of further

H. Zheng, H. Lu / Food Chemistry 128 (2011) 10871093

study. In addition, the percentage retentions of AA in all segments

of asparagus during MW + WB are statistically signicantly higher
than those during WB for inactivating 90% of POD, as shown in
Fig. 6. This result should consider that, on the one hand, the k of
POD inactivation was increased during MW + WB, and on the other
hand, the Ea of AA degradation was also increased after microwave
pre-treatment. Therefore, microwave pre-treatment could alleviate
AA loss and accelerate POD inactivation during water blanching of
asparagus. Soysal and Sylemez (2005) also reported that
microwave heating was more effective for inactivating POD and
preserving ascorbic acid in carrot than thermal treatment. Via
et al. (2007) suggested that microwaves pre-blanching showed
no deleterious effect on ascorbic acid content of Brussels sprouts.
4. Conclusions
AA degradation and POD inactivation in blanched asparagus
followed rst-order kinetics. The bud segment of asparagus should
be blanched to a relatively short period of time for preserving AA
content and inactivating POD activity. In addition, microwaves
may be an effective pre-treatment process for use prior to water
blanching to reduce the degradation of AA and to accelerate the
inactivation of POD and thus maintain produce quality. Future
work will mainly cover the effectiveness of microwave pre-treatment to other fruits and vegetables during water blanching and
their reasons.
Acknowledgements
This research is supported by Postgraduate Innovation Research
Projects of Zhejiang Province (YK2008038). The authors would like
to thank Mr. Heqiang Lou, Ms. Hedan Li and Ms. Lingling Jiang in
the College of Chemistry and Life Science at Zhejiang Normal University (Jinhua, China) for chemical analysis.
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