Journal of Food Engineering 120 (2014) 135145

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Journal of Food Engineering

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

Single and interactive effects of process variables on microwave-assisted

and conventional extractions of antioxidants from vegetable solid
wastes
Antonietta Baiano a, Luisa Bevilacqua a, Carmela Terracone a, Francesco Cont b,
Matteo Alessandro Del Nobile a,
a
b

Dipartimento di Scienze Agrarie, degli Alimenti e dellAmbiente, University of Foggia, Via Napoli 25, 71122 Foggia, Italy
Dipartimento di Economia, University of Foggia, Largo Papa Giovanni Paolo II, 71121 Foggia, Italy

a r t i c l e

i n f o

Article history:
Received 19 February 2013
Received in revised form 15 May 2013
Accepted 14 July 2013
Available online 25 July 2013
Keywords:
Asparagus
Cauliower
Celery
Chicory
Phenolic extraction

a b s t r a c t
The aim of this work was to study the single and interactive effects of process variables in microwaveassisted and conventional extraction of antioxidant compounds from asparagus, cauliower, celery,
and chicory wastes using water as solvent. The following variables were investigated: water/sample ratio,
extraction time in microwave-assisted extraction; water/sample ratio, extraction time, temperature in
conventional extraction. Concerning the microwave-assisted extraction, the highest phenolic recoveries
and the highest antioxidant activity were obtained at a solid-to-liquid ratio of 1:2 (w/w) and prolonging
the time of treatment up to 4 min. In the conventional extraction system, the highest yields were generally obtained at a solid-to-liquid ratio of 1:2 (w/w) and with the combination high temperature-low time
of treatment. Cauliower and chicory wastes showed the highest extraction yields when submitted to
microwaves and conventional extraction, respectively. Catechin, ascorbic acid, and quercetin 3-O-glucopyranoside were the main phenolic compounds identied in wastes. The application of conventional
extraction system gave higher extraction yields than the microwave-assisted one.
2013 Elsevier Ltd. All rights reserved.

1. Introduction
In the last few decades, as a consequence of the increased interest in healthy foods, a growing number of researches have been
performed on the recognition of molecules of plant origin (mostly,
antioxidants) that seem to play a role in the prevention of many
diseases (Hamid et al., 2010; Pandey and Rizvi, 2009; VladimirKnezevic et al., 2012).
Among bioactive substances, polyphenols play an important
role in humans, since they are unable to synthetize these compounds and need to satisfy their phenolic requirements through
a daily consumption of fruits and vegetables. Phenolic compounds
are known to be effective in prevention of chronic and acute diseases such as cancer, cardiovascular disorders, inammations
(Linseisen and Rohrmann, 2008).
Food products obtained from plants are important sources of
phenolic compounds. However, a signicant percentage of the
population in Western and developing countries are not consuming sufcient quantities of dietary polyphenols as a result of inadequate intakes (Martin and Appel, 2010). Furthermore, remarkable

Corresponding author. Tel./fax: +39 881 589242.

E-mail address: ma.delnobile@unifg.it (M.A. Del Nobile).
0260-8774/$ - see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jfoodeng.2013.07.010

amounts of phenolic compounds are lost during processing. Freshcut fruits and vegetables are submitted to a series of technological
operations responsible for physical, enzymatic, and chemical
changes that affect their nutritional and sensorial quality. Operations like sorting, cleaning, washing, peeling and cutting/chopping
can induce different types of stress and parameters such as the
phenolic compounds can suffer noticeable losses (Amarowicz
et al., 2009).
The term minimally processed vegetable is applied to any fresh
vegetable that has been physically altered from its original form,
but remains in a fresh state (Gomez-Lopez et al., 2009). Consumers
are increasingly demanding convenient, ready-to-use, and readyto-eat fruits and vegetables, having fresh-like quality, and containing only natural ingredients. In Europe, particularly in France but
also in the UK, the market for minimally processed fruit and vegetable grew explosively in recent decades (Barry-Ryan et al., 2007).
Since the increasing demand for processed plant products led to a
substantial increase in solid wastes, solutions for their recovery
must be found. In fact, these residues contain bioactive compounds
with potential application in foods, cosmetic and pharmacologic
formulations. Peels, skins, seeds, stems and other lignocellulosic
fractions, usually discarded, are attractive sources of antioxidants
(Laroze et al., 2008). Utilization of by-products could be a source

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A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

of revenue for the company, as these extracts may be sold to pharmaceutical companies and cosmetic manufacturers, or used by
food producers to obtain functional products.
These compounds can be extracted from the vegetable matrix.
Extraction with solvents is the technique most frequently used to
obtain crude extracts of antioxidants of plant origin. The most
common solvents are water, ethanol, methanol, acetone and ethyl
acetate, used pure or in mixture. The structural diversity of antioxidant compounds extracted affects both the returns of extraction
and the activity of the extracts in relation to the solvent used
(Marinova and Yanishlieva, 1997). Also the extraction protocols
are quite different and include conditions ranging from room temperature to boiling or reux. The antioxidant efcacy is related to
the nature of the extracted compounds and also to the type of solvent used. Consequently, for each vegetable matrix, it is necessary
to select solvents and conditions able to maximize yield and the
antioxidant activity of the extracts.
The conventional methods for extraction of antioxidant compounds include uid extraction and soxhlet extraction, which are
used for many decades but have the disadvantages to be very time
consuming and to require relatively large quantities of solvents
(Proestos and Komaitis, 2008). Microwave-assisted methods have
a range of advantages, including: rapidity; reductions in solvent
consumption; better chance of control, automation, and coupling
extraction on-line with analysis of the extracts. Microwave-assisted
extraction also is more environmental friendly, since it requires less
energy and can efciently use a wide range of solvents, including
non-toxic and low environmentally hazardous solvents ones.
The present study was aimed to investigate the single and interactive effects of selected process variables on yield and antioxidant
activity of aqueous extracts obtained through conventional and
microwave-assisted extraction from vegetable solid wastes. The
experiments were focused on matrices such as cauliowers, celery,
chicory and asparagus because the production of the corresponding minimally processed products gives high amounts of solid
waste (an average of 50%). The choice of the variables to be studied
(water/sample ratio, extraction time, and temperature in conventional extraction; water/sample ratio and extraction time in microwave-assisted extraction) depended on their impact on both
efcacy/efciency and energy cost of the process (Spigno et al.,
2007). Authors applied the conventional and microwave-assisted
extraction using water as a solvent because this solvent shows several advantages, including reduction of environmental hazards and
possibility to use the aqueous extracts to fortify bread and other
products whose process include the formation of a dough.
2. Materials and methods

2.3. Extraction systems

Microwave-assisted and conventional extraction systems were
applied according the experimental conditions reported below.
2.4. Microwave-assisted extraction of the antioxidant fraction
The extractions were performed with a domestic microwave
oven (JT 366, 6th sense Whirlpool Europe, Comerio, Italy).
Antioxidants were recovered from the solid waste using water
as a solvent. Three solid-to-water ratio were tested: 1:1, 1:2, and
1:4 (w/w). The nal standard mass was always equal to 300 g.
The resulting mixtures were treated by microwaves at 750 W for
2 or 4 min, according to the methods of Pan et al. (2003) and Proestos et al. (2008). The temperatures of the mixtures were measured
at the end of each treatment through a Pt100 temperature probe.
The mixtures were ltered through a common Whatman paper
and stored at 20 C until analysis.
2.5. Conventional extraction of the antioxidant fraction
The extractions were performed with by using a heating system
consisting of a heating magnetic stirrer, a hemispheric bowl that
allowed an uniform distribution of the heat, a digital thermoregulator, a Pt100 temperature probe (Velp Scientica, Usmate, Italy).
Antioxidants were recovered from the solid wastes using water
as a solvent. Three solid-to-water ratios were tested: 1:1, 1:2, and
1:4 (w/w). The nal standard mass was always equal to 300 g. The
resulting mixtures were treated at a temperature of 50 and 70 C
(the solvent was previously heated to the desired temperature)
respectively for 35 and 20 min. These conditions were selected
after an accurate study of the available literature (Marete et al.,
2009; Spigno et al., 2007). The mixtures were ltered through a
common Whatman paper and stored at 20 C until analysis.
2.6. Determination of the total phenolic content
The total phenolic compounds were determined according to
the FolinCiocalteau method (Gorinstein et al., 2000). In a test tube,
100 mL of phenolic extract or phenolic standard were mixed with
the FolinCiocalteau reagent (2 M, 100 mL) and, after 4 min, with
an aqueous solution of Na2CO3 (5%, 800 mL). The mixture was kept
for 20 min in a water bath set at 40 C, then the total phenol content
was determined colorimetrically at 765 nm. Quantication was
based on a standard curve built with 50100200400600800
1000 mg/L gallic acid (ExtraSynthese, Genay, France) aqueous solution. The total phenolic content was expressed as milligrams of gallic acid equivalents per kilogram of fresh material.

2.1. Plant materials

2.7. HPLC antioxidant prole
The solid wastes originating by the production of ready-to-use
vegetables were supplied by Futuragri Soc. Coop. Agric. (Foggia,
Italy). They included: defective spears and parts of shoots (asparagus, Asparagus ofcinalis, cv. UC 157), external leaves and stems
(cauliower, Brassica oleracea, cv. Vedis e Mildis), external leaves
and stems (chicory, Cichorium intybus L., cv. Puntarelle di Galatina),
external leaves and stems (celery, Apium graveolens, cv. Darklet).
The moisture contents were very high for all the types of solid
wastes. In a decreasing order: celery 92.5%, chicory 92.4%, asparagus 91.8%, and cauliower 89.6%.
2.2. Pretreatment
Before extraction, the solid wastes were nely chopped (5
8 mm-size) by a 30 s-treatment in a domestic cutter (Philips HR
1396/55, Milano, Italy).

The HPLC analysis of the extracts was carried out according to

Yildiz et al. (2008), using a HPLC binary system consisting of a degasser mod. G1322A, a binary pump mod. G1312A, an autosampler
mod. G1329A equipped with a 20-lL loop, and a diode array detector mod. G1315D (Agilent, Santa Clara, CA, USA).
The stationary phase was a Gemini C18 110A analytical column
(250 mm  4.6 mm i.d., 5 lm, Phenomenex, Castelmaggiore, Italy).
The mobile phases for chromatographic analysis were (A) methanol and (B) 0.2% of o-H3PO4 in bidistilled water at constant ow
rate of 1 mL/min. The gradient program of solvent was as follows:
7% A for 8 min, 730% A in 5 min, 3066% A in 35 min, 6675% A in
7 min. Identication of the antioxidant compounds was performed
by comparing the spectra and the relative retention times of the
sample peaks with those obtained by injection of 30 pure standards. Quantication was made on the basis of calibration curves

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A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

Table 1
Microwave-assisted extraction: single and interactive effects of solid-to-liquid ratio and extraction time on antioxidant content and antioxidant activity of cauliower, celery,
chicory, and asparagus extracts. In the last column, the temperatures of the extracts, measured at the end of the extraction, have been reported.
Wastes

Extraction conditions

Cauliower

Solid-to-liquid ratio
1:1 (w/w)
284 b
1:2 (w/w)
468 c
1:4 (w/w)
243 a
Signicance

Extraction time
4 min
360 b
2 min
303 a
Signicance

Solid-to-liquid ratio  extraction time
1:1 (w/w)  4 min
310 24 c
1:1 (w/w)  2 min
257 14 b
1:2 (w/w)  4 min
450 7 d
1:2 (w/w)  2 min
486 8 d
1:4 (w/w)  4 min
321 55 c
1:4 (w/w)  2 min
165 41 a
Signicance


Celery

Chicory

Asparagus

Total phenolic content

(mg gallic acid/kg of
fresh material)

DPPH assay (mmol

Trolox eq/kg of
fresh material)

ABTS assay
(mmol Trolox eq/kg of
fresh material)

0.24 a
0.43 c
0.36 b


19714 a
35736 b
35746 b


0.43 b
0.27 a


36795 b
2449 a


0.31 0.03
0.13 0.03
0.44 0.05
0.41 0.07
0.58 0.10
0.20 0.04


c
a
d
d
e
b

Solid-to-liquid ratio
1:1 (w/w)
106 b
0.07 b
1:2 (w/w)
137 c
0.11 c
1:4 (w/w)
66 a
0.04 a
Signicance


Extraction time extraction time
4 min
116 b
0.11 b
2 min
91 a
0.03 a
Signicance


Solid-to-liquid ratio  extraction time solid-to-liquid ratio  extraction time
1:1 (w/w)  4 min
132 20 e
0.10 0.02 c
1:1 (w/w)  2 min
81 8 c
0.02 a
1:2 (w/w)  4 min
155 13 f
0.16 0.02 d
1:2 (w/w)  2 min
120 21 d
0.02 0.01 a
1:4 (w/w)  4 min
60 5 a
0.04 0.01 b
1:4 (w/w)  2 min
72 7 a,b
0.03 0.01 a,b
Signicance


Solid-to-liquid ratio solid-to-liquid ratio
1:1 (w/w)
169 b
0.04 a
1:2 (w/w)
279 c
0.12 c
1:4 (w/w)
127 a
0.09 b
Signicance


Extraction time extraction time
4 min
261 b
0.11 b
2 min
123 a
0.02 a
Signicance


Solid-to-liquid ratio  extraction time solid-to-liquid ratio  extraction time
1:1 (w/w)  4 min
214 3 e
0.07 0.02 b
1:1 (w/w)  2 min
125 14 b
0.01 a
1:2 (w/w)  4 min
392 32 f
0.21 0.06 d
1:2 (w/w)  2 min
167 11 c
0.01 a
1:4 (w/w)  4 min
176 34 d
0.09 c
1:4 (w/w)  2 min
69 13 a
0.10 b,c
Signicance


Solid-to-liquid ratio solid-to-liquid ratio
1:1 (w/w)
159 b
0.06 c
1:2 (w/w)
210 c
0.02 b
1:4 (w/w)
145 a
0.01 a
Signicance


Extraction Time Extraction Time
4 min
204 b
0.05 b
2 min
138 a
0.02 a
Signicance


Solid-to-liquid ratio  extraction time solid-to-liquid ratio  extraction time
1:1 (w/w)  4 min
195 9 c
0.09 0.02 d
1:1 (w/w)  2 min
122 11 a
0.02 0.01 b,c
1:2 (w/w)  4 min
245 16 d
0.03 0.01 c
1:2 (w/w)  2 min
175 9 b
0.01 0.01 a,b
1:4 (w/w)  4 min
173 11 b
0.01 a
1:4 (w/w)  2 min
117 23 a
0.01 0.01 a,b
Signicance



21354 4280
18074 2553
41568 5566
29070 4428
46361 8132
25890 4363


a
a
c
b
d
b

Temperature at
the end of the
extraction (C)

63
58
68
53
69
56

9668 a
9298 a
11965 b

11166 b
9159 a

10516 3143 b,c
8651 1688 a,b
10554 1926 b,c
6785 2644 a
12925 3104 c
11165 2666 b,c


68
55
74
60
71
52

8262.71 a
23165.03 b
26937.81 c

23869.38 b
10293.09 a

10290 1348 b
6236 684 a
31679 4640 d
9543 1434 b
28092 5256 c
24341 952 c


83
48
82
63
83
55

20082 a
30312 b
39601 c

33982 b
2289 a

20200 2795 a
19955 4099 a
41555 9843 c
21478 2701 a
43120 2645 c
32564 7840 b


In column, within each type of wastes, different letters indicate signicant differences at p < 0.05 by LSD multiple range test.
The asterisks indicate signicant differences at p < 0.05 by LSD multiple range test.

67
52
67
57
71
56

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A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

of the pure standards. Results were expressed as mg per kilogram

of fresh material.

kg of fresh material. The Trolox calibration was built by analyzing

6 methanolic solutions in the range 0.051 mM.

2.8. Evaluation of antioxidant activity by the ABTS
+ assay

2.10. Statistical analysis

The antioxidant activity was measured through the ability of

the extracts to scavenge the ABTS radical cation (reaction followed
by evaluating the reduction of the absorption at 734 nm), according to Re et al. (1999). Briey, ABTS was dissolved in redistilled
water at a concentration of 7 lmol/L. ABTS
+ radical cation was
produced by the reaction of the ABTS stock solution with a
2.45 mmol/L potassium persulfate solution and keeping the
mixture at room temperature in the dark for 16 h. The resulting
solution was diluted with ethanol to an absorbance of 0.70
(0.02) at 734 nm. One milliliter of diluted ABTS
+ solution
(A734nm = 0.70 0.02) was added to 30 lL of the extracts. Then,
the absorbance was taken exactly 4 min after initial mixing (Yang
and Guixing, 2011). The results were expressed as mmol Trolox eq
per kilogram of fresh material.

The extractions of the phenolic compounds were carried out at

least three times and analyses were performed at least in triplicate.
The averages and the standard deviations were calculated using
Excel software ver. 11.5.1 (Microsoft, Redmond, WA). The Analysis
of Variance (ANOVA) at p < 0.05 followed by the LSD test was applied to highlight signicant differences among samples. All the
statistical analyses were made through the software Statistica version 7 (Statsoft, Tulsa, OK). The comparisons between conventional
and microwave-assisted extraction were made between extracts
obtained in conditions that, based on preliminary experimental
tests carried out by the authors, can be considered as equivalent.
In particular, for each solid-to-liquid ratio, the comparisons were
made between the following treatments: 750 W  2 min (microwaves) and 50 C  35 min (conventional); 750 W  4 min (microwave) and the 70 C  20 min (conventional).

2.9. Evaluation of antioxidant activity by the DPPH
assay

3. Results and discussion
The 1,1-diphenyl-2-picrylhydrazyl (DPPH) method was used to
measure the free radical scavenging capacity of extracts
(Brand-Williams et al., 1995). A volume of 1.75 mL of a 6.10 5 M
methanolic solution of the stable organic radical DPPH (DPPH)
was added to 0.25 mL aliquots of phenolic extracts diluted, when
necessary, with known volumes of the extraction solution. The
absorbance at 515 nm was read at regular intervals of time until
the end of the reaction (no changes of the absorbance values in
60 s). Results were expressed as mmol of Trolox equivalents per

3.1. Microwave-assisted extraction

As can be inferred from Table 1 the recoveries of antioxidant
compounds were in a decreasing order: cauliower, chicory, asparagus, and celery. The same tables highlight that the solid-to-liquid
ratio and the time of treatments exerted signicant single and
interactive effects on the amount of antioxidants extracted from
the wastes and the antioxidant activity of the extracts. For all types

Table 2
Concentrations (mg/kg of fresh material) of the phenolic compounds obtained by microwave-assisted extraction.
Ascorbic Catechin Chlorogenic Quercetin 3-Oacid
acid
glucopyranoside

Chicoric Hesperidin Myricetin Quercetin 3acid

O-rutinoside

Isorhamnetin 3- Gallic Caffeic Rosmarinic

O-rutinoside
acid
acid
acid

Cauliower
1:1 (w/w)  4 min

86 5 d

1:1 (w/w)  2 min

23 5 b

91 56
c
62 a

1:2
1:2
1:4
1:4

15 6 b
40 4 c
a
18 5 b

22 6
15 6
18 7
15 2

b 61 b
b 72 b
b 82 b
b
a

23 5
b
21 a
2a
21 a
21 a

a
31
62
3a
71
81

a
a
13 2 b
a
a
a

a
21
31
21
52
a

a
a
a
b

a
a
a
a
a
81 b

a
a
a
a
10 1 b
a

10 1 c
62 b
10 3 c
a
a
a

15 6 b
a
a
a
a
a
a
a
a
6b
a
53 b

a
a
a
62 b
a
a

a
56 8 b
a
71 15 b.c
83 20 c
92 19 c

Solid-to-liquid
ratio  extraction
time

(w/w)  4 min
(w/w)  2 min
(w/w)  4 min
(w/w)  2 min

Celery
1:1 (w/w)  4 min
1:1 (w/w)  2 min
1:2 (w/w)  4 min
1:2 (w/w)  2 min
1:4 (w/w)  4 min
1:4 (w/w)  2 min

Chicory
1:1 (w/w)  4 min
1:1 (w/w)  2 min
1:2 (w/w)  4 min
1:2 (w/w)  2 min
1:4 (w/w)  4 min
1:4 (w/w)  2 min

24 3 b
a
a
a
a
a

61
62
a
52
92
73

Asparagus
1:1 (w/w)  4 min
1:1 (w/w)  2 min
1:2 (w/w)  4 min
1:2 (w/w)  2 min
1:4 (w/w)  4 min
1:4 (w/w)  2 min

a
b
b
a
b
b
b
b
b

48 7 c

In column, within each type of solid wastes, different letters indicate signicant differences at p < 0.05.

7 3 b 10 2 b
a

a
a
a
a

a
a
a
a

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A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

Table 3
Conventional extraction: single and interactive effects of solid-to-liquid ratio and extraction time and temperature on antioxidant content and antioxidant activity of cauliower,
chicory, celery, and asparagus extracts. In the last column, the temperatures of the extracts, measured at the end of the extraction, have been reported.
Wastes

Extraction conditions

Cauliower

Solid-to-liquid ratio
1:1 (w/w)
408 b
1:2 (w/w)
346 a
1:4 (w/w)
342 a
Signicance

Extraction timetemperature
70 C 20 min
437 b
50 C 35 min
289 a
Signicance

Solid-to-liquid ratio  extraction timetemperature
1:1 (w/w)  70 C 20 min
529 23 c
1:1 (w/w)  50 C 35 min
262 22 a
1:2 (w/w)  70 20 min
440 57 b
1:2 (w/w)  50 C 35 min
252 34 a
1:4 (w/w)  70 C 20 min
335 83 a
1:4 (w/w)  50 C 35 min
407 7 b
Signicance


Celery

Chicory

Asparagus

Total phenolic content

(mg gallic acid/kg of
fresh material)

Solid-to-liquid ratio
1:1 (w/w)
112 a
1:2 (w/w)
200 b
1:4 (w/w)
101 a
Signicance

Extraction timetemperature
70 C 20 min
177 b
50 C 35 min
96 a
Signicance

Solid-to-liquid ratio  extraction timetemperature
1:1 (w/w)  70 C 20 min
139 13 b
1:1 (w/w)  50 C 35 min
57 1 a
1:2 (w/w)  70 20 min
290 67 c
1:2 (w/w)  50 C 35 min
109 9 b
1:4 (w/w)  70 C 20 min
100 25 b
1:4 (w/w)  50 C 35 min
102 1 b
Signicance

Solid-to-liquid ratio
1:1 (w/w)
434 a
1:2 (w/w)
831 b
1:4 (w/w)
490 a
Signicance

Extraction timetemperature
70 C 20 min
673 b
50 C 35 min
497 a
Signicance

Solid-to-liquid ratio  extraction timetemperature
1:1 (w/w)  70 C 20 min
656 32 c
1:1 (w/w)  50 C 35 min
212 14 a
1:2 (w/w)  70 20 min
868 219 d
1:2 (w/w)  50 C 35 min
794 190 c.d
1:4 (w/w)  70 C 20 min
494 51 b
1:4 (w/w)  50 C 35 min
486 17 b
Signicance

Solid-to-liquid ratio
1:1 (w/w)
220 a
1:2 (w/w)
624 c
1:4 (w/w)
392 b
Signicance

Extraction timetemperature
70 C 20 min
387 a
50 C 35 min
439 b
Signicance

Solid-to-liquid ratio  extraction timeTemperature
1:1 (w/w)  70 C 20 min
243 7 b
1:1 (w/w)  50 C 35 min
197 15 a
1:2 (w/w)  70 20 min
557 37 e
1:2 (w/w)  50 C 35 min
690 38 f
1:4 (w/w)  70 C 20 min
360 18 c
1:4 (w/w)  50 C 35 min
430 44 d
Signicance


DPPH assay (mmol

Trolo  eq/kg of
fresh material)

ABTS assay (mmol

Trolox eq/kg of
fresh material)

0.32 b
0.25 a
0.26 a


25177 a
26071 a
36267 b


0.40 b
0.11 a


36880 b
23054 a


0.44 0.04
0.13 0.02
0.38 0.05
0.09 0.01
0.37 0.02
0.10 0.02


d
b
c
a
c
a,b

28801 5547 b
20586 4182 a
46224 12078 c
18959 3550 a
45000 8900 c
29718 4403 b


0.07 b
0.08 b
0.05 a


5015 a
14239 c
6635 b


0.09 b
0.03 a


11148 b
4000 a


0.09 0.02
0.02 a
0.13 0.04
0.02 0.01
0.04 0.01
0.06 0.01


c
d
a
b
b

6231 1216 b
3314 635 a
21035 4789 c
2589 504 a
6997 2011 b
6093 1189 b


0.34 b
0.04 a
0.04 a


41429 b
15063 a
12781 a


0.25 b
0.04 a


35207 b
12777 a


0.58 0.10
0.03 0.01
0.04 0.01
0.04 0.01
0.05 0.02
0.04 0.01


b
a
a
a
a
a

58597 1095 c
9735 2098 a
11085 1958 a
18295 3513 b
16534 3567 b
9528 1986 a


0.09 b
0.06 a
0.08 b


10736 a
14044 b
21646 c


0.10 b
0.02 a


17788 b
12508 a


0.12 0.02
0.04 0.01
0.10 0.01
a
a
0.12 0.02


c
b
c

In column, within each type of wastes, different letters indicate signicant differences at p < 0.05 by LSD multiple range test.
The asterisks indicate signicant differences at p < 0.05 by LSD multiple range test.

14330 2927 b
7441 2217 a
18237 3328 c
10231 1922 a
20582 3862 c,d
2341 6852 d


140

A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

of wastes, the highest recoveries of phenolics were obtained with a

solid-to-liquid ratio equal to 1:2 (w/w) and a time of treatment of
4 min. The highest values of antioxidant activity were detected in
the same conditions (solid-to-liquid ratio 1:2 and 4 min) with
the exception of the values referred to the ABTS assay applied to
celery and asparagus and of those concerning the DPPH assay applied to the asparagus. The increase of extraction yields from 1:1
to 1:2 (w/w) solid-to-liquid ratio was consistent with literature
and with the principle of the mass transfer, whose driving force
is represented by the concentration gradient between solid and liquid phase (Radojkovic et al., 2012). This gradient increases as the
solid-to-liquid ratio decreases down to a limit value beyond which
the extraction yield does not increase. The reduction of the phenolic recovery detected with the further decrease of the solid-to-liquid ratio from 1:2 to 1:4 (w/w) was also consistent with
literature. It could be explained with the change of the activity
coefcients (and thus of the solubility of the antioxidant compounds), which is a consequence of the change of the composition
of the solution and of the interactions between extracted compounds and solvent (Tan et al., 2011; Frank et al., 1999). Concerning the extraction time, it is of primary importance to minimize
energy and cost of the extraction process. Nevertheless, according
to previous works, the phenolic concentration of extracts increased
with time of contact, as explained by the Ficks second law of diffusion. Furthermore, as highlighted by Table 1, and as one would

expect, the temperatures measured at the end of the microwave

treatments increased as time of treatment increased and, consequently, the increase of temperature had a further positive effect
on the extraction yields (Bucic-Kojic et al., 2011). These results also
suggested that phenolics from solid wastes of asparagus, cauliower, celery, and chicory and the corresponding antioxidant
activity are relatively stable under high temperature conditions.
They also conrm the ndings of Liazid et al. (2007), who studied
the effects of microwave irradiation on the stability of over 20 phenolic compounds, founding that all of them were stable for up to
20 min at 100 C.
As reported above, temperature increased as time of treatment
increased. Furthermore, temperature was not affected by the solidto-liquid ratio since the total mass did not change. Concerning the
4 min treatment, the highest mean temperatures (82 C) were detected on chicory wastes. The mean temperature measured on the
other wastes at the end of that treatment was equal to 69 C. No
differences were detected between wastes in the case of the
2 min-treatment since the mean temperature detected was always
equal to 55 C. Concerning the phenolic proles (Table 2), 4, 4, 6,
and 1 compounds were identied respectively in cauliower, celery, chicory, and asparagus extracts. According to the data of Table 2, the molecules detected in higher concentrations were:
ascorbic acid in cauliower and chicory, catechin in celery extracts,
and quercetin 3-O-rutinoside in asparagus extracts.

Table 4
Concentrations (mg/kg of fresh material) of the phenolic compounds obtained by conventional extraction.
Solid-to-liquid
ratio  extraction time
temperature time
Cauliower
1:1 (w/w)  70 C 20 min

Ascorbic
acid

1:1 (w/w)  50 C 35 min

361 139 25 12 21 11 c
d
a,b
a
14 6 a
a

1:2 (w/w)  70 20 min

198 2 c

1:2 (w/w)  50 C 35 min

94 59 b

1:4 (w/w)  70 C 20 min

367 161 43 11 15 7 b.c

d
b
116 58 b 39 7 b
a

22 11 b

1:4 (w/w)  50 C 35 min

Celery
1:1 (w/w)  70 C 20 min
1:1 (w/w)  50 C 35 min
1:2 (w/w)  70 20 min

Chicory
1:1 (w/w)  70 C 20 min
1:1
1:2
1:2
1:4
1:4

(w/w)  50 C 35 min
(w/w)  70 20 min
(w/w)  50 C 35 min
(w/w)  70 C 20 min
(w/w)  50 C 35 min

Asparagus
1:1 (w/w)  70 C 20 min
1:1 (w/w)  50 C 35 min
1:2 (w/w)  70 20 min
1:2 (w/w)  50 C 35 min
1:4 (w/w)  70 C 20 min
1:4 (w/w)  50 C 35 min

29 1
a.b
20 a

11 2 b
a

15 3 b 12 3 b
51 a
a
40 16 19 4 c
d
81 a
a
22 2 c
a
15 1 b
a

1:2 (w/w)  50 C 35 min

1:4 (w/w)  70 C 20 min
1:4 (w/w)  50 C 35 min

Catechin Chlorogenic Quercetin 3-O- Chicoric Coumaric Myricetin Apigenin Isorhamnetin Gallic Caffeic Quercetin
acid
acid
3-O
acid
glucopyranoside acid
acid
3-Orutinoside
rutinoside

77 33 b
a
a
a
a

72 b
10 2 b
a
a
a

a
a
a
a
a

a
a
a

83
b
42
a
a

42
a
a

81
b

14 1 b
a
41 16 c

61b

a
63 b
52 b

3b
a
3b
a
3b

10 01
b
a
a
a
a
a

In column, within each type of solid wastes, different letters indicate signicant differences at p < 0.05.

a
a
a

a
a
a
a
a

83 17 a
77 14 a
104 b
120 8 c
123 19 c
131 24 c

A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

141

Fig. 1. Statistical comparison between the two extraction methods in terms of total phenolic yield. Comparison between: (a) the 750 W  2 min-microwave assisted
treatment and the 50 C  35 min conventional one; (b) the 750 W  4 min-microwave assisted treatment and the 70 C  20 min conventional one different letters indicate
signicant differences at p < 0.05 by LSD multiple range test.

3.2. Conventional extraction

According to Table 3, the highest recovery of antioxidant compounds was obtained for chicory, followed by asparagus, cauliower, and celery. Also in the case of the conventional
extraction, the solid-to-liquid ratio and the time of treatments exerted signicant single and interactive effects on both the amount
of antioxidant compounds extracted from the wastes and the antioxidant activity of the extracts. However, these effects were specic for each of the considered vegetable wastes.
The highest recovery of antioxidant compounds was obtained
with treatments performed at a solid-to-liquid ratio of 1:2 (w/w),
at 70 C for 20 min. The only exception was represented by the
asparagus wastes whose highest extraction yields were obtained
by heating at 50 C for 35 min.
Data from Table 3 highlight that the effects of solid-to-liquid
ratio, time, and temperature of treatment on the antioxidant

contents of the extracts were not always consistent and often statistically not signicant. The highest extraction yields were observed when the combination high temperature-low time of
treatment was applied. This result can be explained by the increase
of the diffusion coefcient and, as a consequence, by the increase of
the diffusion rate of phenolic compounds from the solid to the liquid phase (Jokic et al., 2010).
Concerning the antioxidant activity measured according to the
DPPH assay, the highest values were generally detected on the extracts obtained with treatments performed at a solid-to-liquid ration of 1:1 (w/w), at 70 C for 20 min. Instead, antioxidant values
measured according to the ABTS assay depended on the specic
type of waste.
Concerning the antioxidant proles (Table 4), 6, 4, 7, and 1 compounds were identied respectively in cauliower, celery, chicory,
and asparagus extracts. The molecules detected in higher concentrations and in most of the samples were: ascorbic acid and

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A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

catechin in cauliower; catechin in celery and chicory; quercetin

3-O-rutinoside in asparagus extracts.
3.3. Comparison between the two extraction systems
It can be of interest to compare the microwave-assisted extraction with the conventional one in terms of total phenolics extracted (Fig. 1). The two extraction systems generally gave
different extraction yields from the same vegetable waste. In particular, for each value of the solid-to-liquid ratio, the statistical
comparisons were made between the 750 W  2 min-microwave
assisted treatment and the 50 C  35 min-conventional one
(Fig. 1a), and between the 750 W  4 min-microwave assisted
treatment and the 70 C  20 min-conventional one (Fig. 1b). As
highlighted by Fig. 1, the extraction yields were higher when the
conventional system was applied in most of the operating conditions. Only in few cases there were no signicant differences between the two extraction methods. These results are in contrast
with the ndings of Liazid et al. (2007) and Inglett et al. (2010),

who highlighted the ability of microwaves to destroy the cellular

structure of the vegetable matrix in a more effective way than
the conventional heating, and to release greater amounts of phenolic compounds from the bounded fraction. Durmaz et al. (2012), instead, stated that there was no signicant difference between
microwave and conventional extraction in terms of phenolic content and antioxidant activity when optimum conditions were compared. Nevertheless, data are in agreement with those of Biswas
et al. (2012), who found that conventional heat extractions can
be more efcient than the microwave-assisted one in extracting
antioxidants from beans, and with those of Waksmundzka-Hajnos
et al. (2007), who highlighted the better performance of a soxhlet
apparatus. This minor efciency of the microwave-assisted extraction could be due to inadequate stirring of the solvent by the
microwaves (Eskilsson et al., 1999). Furthermore, the dielectric
properties of the solvent inuence the irradiation time optimization. Longer exposure with microwave absorbing solvents like
water may reduce the concentrations of compounds sensitive to
heat (Afoakwah et al., 2012). Unlike classical conductive heating

Fig. 2. Statistical comparison between the two extraction methods in terms of antioxidant activity measured through the DPPH assay. Comparison between: (a) the
750 W  2 min-microwave assisted treatment and the 50 C  35 min-conventional one; (b) the 750 W  4 min-microwave assisted treatment and the 70 C  20 minconventional one Different letters indicate signicant differences at p < 0.05 by LSD multiple range test.

A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

methods, microwaves heat the whole sample simultaneously and

the advantage of microwave-assisted extraction is the disruption
of weak hydrogen bounds promoted by the dipole rotation of the
molecules. Nevertheless, a higher viscosity of the medium lowers
this mechanism by affecting molecular rotation and this effect
could explain our results, in particular when the 1:1 (w/w) solidto-liquid ratio was applied. In any case, microwave-assisted
extraction is able to extract phenolic compounds more quickly
than the conventional procedure. Previous researches demonstrated that yield depends primarily on the plant material from
which the phenolic fraction was extracted and that an extraction
method can be better than another one depending on the vegetable matrix (Camel and Bermond, 1999).
According to DPPH assay (Fig. 2), the antioxidant activity values
of microwave and conventional extracts did not follow a general
rule while the ABTS assay (Fig. 3) show higher values in most of
the extracts obtained through microwaves. DPPH measure the
overall radical-scavenging activity of a sample since it is not
specic to any particular antioxidant component while ABTS is

143

generally used to screen the relative radical-scavenging abilities

of phenolics. Thus, in the current operating conditions, even if
the extracts obtained through microwaves had lower phenolic contents than the conventional extracts, microwaves preserved the
antioxidant activity more effectively than the conventional extraction. This behavior can be explained by the nding that time of
heating is another important factor that inuences the microwave-assisted extraction process. It is well known that the amount
of analyte extracted can be increased by increasing the extraction
time but, in microwave assisted extraction, there is an associated
risk of degradation of thermolabile components (Al-Harahsheh
and Kingman, 2004) due to the already cited prolonged exposure
with microwave absorbing solvents.
The statistical comparisons of data from Tables 3 and 4 are not
reported, nevertheless the two extraction systems affected in a different way also the antioxidant proles as a consequence of the
different mechanisms of action that they exerted on the vegetable
tissues and the different distribution of compounds between the
free and conjugated phenolic fraction. In fact, most of the phenolic

Fig. 3. Statistical comparison between the two extraction methods in terms of antioxidant activity measured through the ABTS assay. Comparison between: (a) the
750 W  2 min-microwave assisted treatment and the 50 C  35 min-conventional one; (b) the 750 W  4 min-microwave assisted treatment and the 70 C  20 minconventional one Different letters indicate signicant differences at p < 0.05 by LSD multiple range test.

144

A. Baiano et al. / Journal of Food Engineering 120 (2014) 135145

compounds are present in conjugated forms, mainly bound to sugars. It is well known that the type of conjugation affects solubility
of phenolic compounds, which can be found in a soluble or in an
unsoluble bound form and their proportion is different in the different investigated vegetable tissues. In fact, according to the
Database on polyphenol content in foods (http://www.phenol-explorer.eu): cauliower contained avonol and avone derivatives
that can be extracted by acid hydrolysis; avones can be obtained
from celery, always by acid hydrolysis; chicory contains free phenolic acid and avones and avonols present as both glycosides
and glucuronides; and asparagus contain glucosides and phenolic
acids in form of ester and ether linked to the cell wall and released
by alkaline hydrolysis. An interesting data is represented by cauliower ascorbic acid contents, which were in reversed trend between microwave-assisted and conventional extraction methods.
In fact, the extraction yield decreased with time of treatment in
microwave extraction while increased with time in the conventional one, even though the extraction temperatures were similar.
These peculiar trends were in agreement with the already cited
ndings of Al-Harahsheh and Kingman (2004) and Afoakwah
et al. (2012).

4. Conclusion
On the basis of the experimental evidences, the higher extraction yields were generally obtained by means of the conventional
system. This statement represents a strong difference respect to
previous works that highlighted, among the advantages of the
microwave-assisted extraction, the higher extraction efciency.
In the case of microwave-assisted extraction, the highest antioxidant recoveries and the highest antioxidant activity were obtained with a solid-to-liquid ratio equal to 1:2 (w/w) and
prolonging the treatment up to 4 min. The effects of both solidto-liquid ratio and time of treatments were statistically signicant.
The cauliower wastes showed the highest extraction yields when
treated by microwaves.
In the conventional extraction system, the highest yield were
generally obtained applying a solid-to-liquid ratio equal to 1:2
(w/w) and the combination high temperature-low time of treatment. In fact, the high temperatures promoted the extraction of
the compounds responsible for antioxidant activity by improving
diffusion phenomena. The highest antioxidant activity measured
according to the DPPH assay was detected in the extracts obtained
in conditions such as: solid-to-liquid ratio equal to 1:1 (w/w), temperature 70 C, time 20 min. Among wastes, chicory showed the
highest extraction yields when treated according to the conventional system.
Summarizing these results, it is possible to state that the optimum parameters and conditions for microwave-assisted extraction were solid-to-liquid ratio 1:2 (w/w) and time of treatment
4 min at 750 W, while conventional extraction generally gave the
best performance at a solid-to-liquid ratio of 1:2 (w/w), temperature 70 C, time 20 min.
Ascorbic acid, catechin, and quercetin 3-O-glucopyranoside
were the main antioxidant compounds detected.

Acknowledgements
One of the authors, Dr. Luisa Bevilacqua, wants to express their
gratitude to REGIONE PUGLIA-P.O PUGLIA FSE 2007-2013 RITORNO AL FUTURO-BORSE DI RICERCA (Avviso 19/2009) for its nancial support.

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