Combined Vacuum Impregnation and

Electron-Beam Irradiation Treatment to Extend

the Storage Life of Sliced White Button
Mushrooms (Agaricus bisporus)
Zeynep Sevimli Yurttas, Rosana G. Moreira, and Elena Castell-Perez

Abstract: This study assessed the application of an antibrowning solution using vacuum impregnation (VI) and then
electron-beam irradiation as a means to extend the shelf life of sliced white button mushrooms (Agaricus bisporus). A
preliminary study helped to determine the best antibrowning solution and VI process parameters. Mushroom slices were
impregnated with 2 g/100 g ascorbic acid + 1 g/100 g calcium lactate; 2 g/100 g citric acid + 1 g/100 g calcium lactate;

E: Food Engineering &

1 g/100 g chitosan + 1 g/100 g calcium lactate; and 1 g/100 g calcium lactate at different vacuum pressures and times and

Physical Properties
atmospheric restoration times. Selection of the antibrowning solution and VI parameters was based on texture and color
of the mushroom slices. Next, the slices were irradiated at 1 kGy using a 1.35-MeV e-beam accelerator. Physicochemical,
sensory, and microbial quality of mushrooms was monitored for 15 d at 4 ◦ C. The best impregnation process in this study
was 2 g/100 g ascorbic acid and 1 g/100 g calcium lactate at 50 mm Hg for 5 min and an atmospheric restoration time
of 5 min. The control (untreated) samples suffered structural losses throughout storage. Only the vacuum impregnated-
irradiated samples had acceptable color by the end of storage. Sensory panelists consistently preferred the samples produced
with VI and irradiation because exposure to ionizing radiation inhibited growth of spoilage microorganisms.

Keywords: dose uniformity ratio, dosimetry, quality, spoilage

Practical Application: Extending the shelf life of mushrooms is important for their marketing and distribution and
reliable preservation methods are still needed. Treating sliced mushrooms with vacuum impregnation and electron-beam
irradiation introduces physiologically active components such as beneficial antioxidants, vitamins, and cations, while
assuring safety and maintaining quality.

Introduction However, these techniques have associated drawbacks including

White button mushrooms have an impact on treatment and
prevention of breast cancer, mainly in postmenopausal women;
they seem to inhibit the activity of aromatase, an enzyme involved
in estrogen production (Teichmann and others 2007; Wani and
others 2009).
The mushroom industry presents about 5% to 25% of its fresh
produce as slices and consumer demand for ready-to-use vegeta-
bles has increased the market for sliced mushrooms (Brennan and
Gormley 1998). However, it is challenging to maintain their qual-
ity because of the lack of cuticle to protect them. Their larger
surface area broadens the spoilage problems. Some critical changes
include browning and softening (Brennan and Gormley 1998;
Sommer and others 2010).
Preservation of mushrooms has been attempted using packag-
ing (Lopez-Briones and others 1992), chemicals (Sapers and others
2001), washing (Cliffe-Byrnes and O’Beirne 2008), tyrosine in-
hibitors (Singh and others 2010), ozone (Yuk and others 2007),
and irradiation (Roy and Bahl 1984; Beaulieu and others 2002).
MS 20130805 Submitted 6/14/2013, Accepted 9/26/2013. Authors are with effects such as texture loss due to tissue softening (Koorapati and
Dept. of Biological and Agricultural Engineering, Texas A&M Univ., College Sta- others 2004; Niemira and Fan 2009; Moreira and Castell-Perez
tion, TX 77843-2117, U.S.A. Direct inquiries to author Castell-Perez (E-mail:
ecastell@tamu.edu).
safety, discoloration, and off-flavors production, and may be un-
suitable for use on an industrial scale (Duan and others 2010).
In the last decades, vacuum impregnation (VI) has been used
as a means to introduce liquids into porous foods. This technique
alters the product’s composition, physical, and chemical properties
to improve its nutritional value (Fito and others 2001; Hironaka
and others 2011). Furthermore, VI of a coating solution may be
an alternative to improve the dispersion retention and to form a
thicker, more effective coating. Vargas and others (2009) observed
that chitosan-based edible coatings applied to fresh-cut carrots by
VI enhanced all the positive effects of the coating.
Sometimes, effective preservation of fresh-cut products may be
achieved using a combination of several treatments (Garcia and
Barrett 2002). Irradiation using gamma rays and electron beams
maintains the produce quality and enhances shelf life in 2 ways:
it kills spoilage organisms and retards plant ripening (Moreira and
Castell-Perez 2012; Mami and others 2013). The recommended
dose for enhancing the shelf life of mushrooms in different coun-
tries ranges from 1 to 3 kGy (Akram and Kwon 2010). However,
irradiation at doses greater than 1 kGy can induce negative quality
2012). Thus, the need to explore the synergistic effect of VI and
irradiation in extending the shelf life of fresh-cut mushrooms.



C 2013 Institute of Food Technologists
R

doi: 10.1111/1750-3841.12308 Vol. 79, Nr. 1, 2014 r Journal of Food Science E39
Further reproduction without permission is prohibited
 Fresh-cut mushroom shelf-life extension . . .

The objectives of this study were to (1) determine the best VI groups without any treatment served as controls. The impregnated
setup in terms of mushroom color and texture, and (2) evaluate and control samples were placed into plastic containers (Ziploc R,

the effect of VI and e-beam irradiation on the physicochemical, 591 mL, with plastic lids) and stored at 4 ◦ C for analysis at days 0,
microbiological, and sensory quality of fresh-sliced mushrooms. 4, 9, 12, and 15 of storage.
Impregnated liquid fraction (X). Sample weight before
Materials and Methods (W1 ) and after (W2 ) the impregnation treatment was measured
and impregnated liquid fraction (X) values were calculated. The
Experimental design impregnated liquid fraction represents the total external liquid that
In a preliminary study, mushroom slices were vacuum impreg- penetrates into the tissue. It was calculated by the weight differ-
◦
nated (VI) with several solutions. During storage at 4 C, color ence of the samples before and after the treatment (Ortiz and
and texture were monitored and the best conditions (impregnation others 2003) using the following formula:
solution, vacuum pressure and duration, and atmospheric restora-
tion time) determined. The best conditions were then used to W2 − W1
design the next set of experiments, which consisted of 1 set with X= × 100 (1)
W1
3 experiments: (1) irradiation, (2) VI, and (3) VI plus irradiation
E: Food Engineering &

(see Irradiation experiment section). Ten slices were used for each treatment with 8 replications. Due
Physical Properties

Four groups of samples were used for product quality and shelf to the sheer amount of samples required, the change in mushroom
life analyses: (1) control (fresh samples without any treatment), (2) composition was calculated after the impregnation process only
impregnated, (3) irradiated, and (4) impregnated-irradiated slices. after determining the most effective VI process. USDA food com-
The experimental design was a 4 × 4 × 2 experiment consisting position values were used as reference (USDA 2011). The change
of 3 factors: impregnating solution (4 different solutions), vacuum in water content after impregnation was confirmed by measuring
pressure (50, 75, 100, and 125 mm Hg), pressure duration (5 and the moisture content of the control and the selected impregnated
10 min), and atmospheric pressure restoration (5 and 10 min). The sample.
experiment was conducted in duplicate. Moisture content. Moisture content of impregnated (ascor-
bic acid + calcium lactate) and control slices were determined by
Sample preparation weight loss after drying in a vacuum oven at 70 ◦ C until con-
Locally grown button mushrooms (Agaricus bisporus) were stored stant weight (AOAC 1990). The samples were randomly chosen.
at 10 ◦ C and 95% relative humidity in plastic bags, washed under Each sample’s weight was recorded before and after drying. More-
tap water, dried with absorbent paper, and sliced (Farberware, over, the weight of canisters was recorded for higher accuracy.
Hillsboro, Tex., U.S.A.). The head sides and long stems where The samples (cap and stem) were first chopped into small pieces
discarded. The equipment (slicer, knife, beakers, and strainers) (approximately 10 g) and then placed in aluminum canisters prior
was sanitized using a 300 μL/L chlorine solution. Slices thickness the drying process. After removal from the vacuum oven, the sam-
was 6.5 ± 0.3 mm with a 3 to 5 mm cap length. ples were placed in a desiccator before recording the final weight.
Measurements were carried out at room temperature in triplicate
Preparation of impregnation (antibrowning) solutions for the control and the selected treatment.
Antibrowning solutions. To prepare the chitosan solution,
0.5 g/100 g acetic acid (Glacial, Mallinckrotd Baker Inc., Paris, Electron-beam irradiation
Ky., U.S.A.) and 1 g/100 g calcium lactate pentahydrate (Sigma- Uniform dose distribution within a sample is important when
Aldrich, St. Louis, Mo., U.S.A.) was dissolved in distilled wa- designing irradiation treatments for food. Uniformity is described
ter at room temperature. Next, chitosan (medium molecular by a low dose uniformity ratio (DUR), the ratio between the
weight, Sigma-Aldrich) was added at 1 g/100 g concentration maximum and minimum dose absorbed by the sample. Although
(Hernandez-Munoz and others 2006). Ascorbic acid (L-Ascorbic the ideal DUR should be close to 1, accepted DUR values for
acid, Sigma-Aldrich) and citric acid (Citric acid Anhydrous, Fisher practical applications are between 1.5 and 2 (IAEA 2002). A dose
Scientific, Fair Lawn, N.J., U.S.A.) solutions were prepared by dis- mapping study was conducted using an ion farm chamber to deter-
solving 2 g/100 g of each acidulant and 1 g/100g calcium lactate mine dose uniformity of the irradiation setup. Irradiation dosage
pentahydrate (Sigma-Aldrich) in distilled water at room temper- was measured by placing Radiochromic film dosimeters (Far West
ature. Similarly, 1 g/100g of calcium lactate pentahydrate was Technology Inc., Goleta, Calif., U.S.A.) at the front, center, and
dissolved in distilled water at room temperature. backside of the mushroom slices, for a total of 3 dosimeters.
Preliminary study. The best impregnation solution, time, and The radiochromic films were read after stabilization using a Ra-
pressure combinations were determined by monitoring color and diochromic reader model 92 (Far West Technology Inc.). The
texture during 15 d at 4 ◦ C. DUR in this study was 1.44, ensuring uniform dose distribution
Impregnation procedure. A VI system composed of a vac- within the mushroom slice. The entrance dose was approximately
uum pump (Emerson Motor Div., St. Louis) and a vacuum glass 0.45 kGy, maximum dose was approximately 0.65 kGy, and back-
desiccator (Pyrex R , Brazil) was used. Sliced mushrooms were im- dose was 0.6 kGy for an applied dose of 0.5 kGy. For a target of
mersed in beakers (approximately 13 slices per beaker, a strainer 1 kGy (this study), the same relationship was applied, with en-
used to keep them immersed) containing the different impreg- trance, maximum and backdoses of 0.90, 1.30, and 1.2 kGy, re-
nation solutions (ascorbic acid, citric acid, calcium lactate, and spectively (DUR = 1.44.) It should be noted that sample thickness
chitosan), 1 solution at a time. During the vacuum step, different was reduced to 3.91 ± 0.16 mm because the DUR was too high
pressures (50, 75, 100, and 125 mm Hg) were applied for 5 and with samples 6.5 mm thick. This sample thickness was used for
10 min and afterward, the atmospheric pressure was restored for 5 the shelf life study.
or 10 min. The impregnated samples were then drained and ex- Irradiation experiment. A group of 4 different samples
cess liquid was removed from the surface with a paper towel. The was evaluated to determine the best treatment: (1) control (no

E40 Journal of Food Science r Vol. 79, Nr. 1, 2014

Fresh-cut mushroom shelf-life extension . . .

treatment), (2) impregnated (2 g/100 g ascorbic acid + 1
g/100 g calcium lactate), (3) irradiated, and (4) impregnated-
irradiated slices. Samples were prepared using the same proce-
dure described above. Prior to irradiation, a single slice was
placed in a plastic bag (Mylar) (Zip SealTM , 8.64 × 10.16 cm,
48GaPET/PE/0.00035Foil/LLDPE, Transilwrap Co., Franklin
Park, Ill., U.S.A) and sealed. The slices were irradiated at 1 kGy,
the maximum allowed by the FDA for fresh produce. We did
not irradiate at lower or higher doses because it has been proven
ineffective (Sapers and others 1994; Koorapati and others 2004).

Table 1–Effect of impregnation treatment (ascorbic acid and citric acid + calcium lactate) on texture (firmness) of sliced mushrooms
impregnated at different vacuum pressures and different atmospheric restoration times during storage at 4 ◦ C.

Time 50 mm Hg 50 mm Hg 75 mm Hg 75 mm Hg 100 mm Hg 100 mm Hg 125 mm Hg 125 mm Hg

(days) Control –5 min –10 min –5 min –10 min –5 min –10 min –5 min –10 min
2 g/100 g ascorbic acid + 1 g/100 g calcium lactate solution (maximum force in newton)

0 a b,c,d b,c a,d a c b,c b a

w 30.297 w 20.05 w 18.846 w 25.19 x 26.74 w 15.562 w 18.76 w,x 19.244 w,x 28.127
1 (2.092) (1.663) (1.562) (0.756) (0.559) (0.686) (1.473) (0.864) (0.394)
4 b a b b c a a a 3b,c
w 28.257 w 20.346 y 30.552 w,x 28.274 y 35.33 x 22.342 w,x 21.585 x 23.162 w,x 32.09
(1.010) (1.904) (2.270) (1.568) (1.938) (1.271) (2.423) (1.809) (1.920)

E: Food Engineering &

9 h a,b a b,c c,g c,f a,c,e a,c,d b,d,e,f,g
y 47.79 x 25.787 x 24.748 x 29.39 y 35.59 y 35.027 y 32.287 y 30.505 x 35.86

Physical Properties
(1.237) (1.020) (1.337) (0.871) (1.548) (2.822) (2.770) (2.950) (3.226)
12 f e,f b,c,d a,b d,e a,b,c a c,d,e b,c,d
x 42.753 y 39.704 y 32.816 w,x 29.05 y 35.616 y 29.353 x 25.236 y 35.1 w,x 32.53
(2.409) (1.924) (1.331) (1.836) (1.998) (0.750) (2.594) (3.910) (1.904)
15 e a b b,c,e c a a a a,b,c
x 43.185 w 20.122 y 32.97 y 34.246 x 25.027 x 20.805 w 18.457 w 18.032 w 25.727
(2.315) (0.751) (1.008) (2.413) (1.510) (1.430) (1.858) (0.978) (2.908)
2 g/100 g citric acid + 1 g/100 g calcium lactate solution (maximum force in newton)
0 w 30.297a x 23.881
a
x 31.328
c,d
x,y 35.573
d
x 29.297
b,c
x 26.127
a,b
x 31.145
b,c,d
x 28.87
a,b,c
x 30.116
b,c
1 (2.092) (2.177) (2.655) (2.039) (2.495) (2.036) (1.906) (2.323) (2.243)
4 b b c b c e d c f
w 28.257 y 35.040 y 49.355 x 32.613 y 48.436 y 61.34 y 53.315 y 46.656 z 67.543
(1.010) (1.897) (1.351) (1.952) (1.559) (0.789) (1.858) (0.800) (1.174)
9 h a,b b,c c c b,c a,b a a
y 47.79 y 38.500 z 43.705 y 46.236 y 46.32 z 43.385 z 37.55 x 34.623 x,y 34.83
(1.237) (2.116) (0.106) (3.136) (0.494) (0.007) (0.961) (1.773) (2.851)
12 f a,b b a,b a,b a,b a,b a,b a,b
x 42.753 x,y 47.840 x,z 24.820 x,y 50.545 x,y 45.77 x,y,z 39.86 x,y,z 58.775 z 55.805 y 39.31
(2.409) (10.534) (1.796) (3.500) (4.341) (3.464) (7.177) (3.655) (3.705)
15 e a a,b a,b a,b a,b a,b a b
x 43.185 y 45.723 x,y,z 33.96 x,y 28.415 x,y 52.46 x,y,z 26.36 x,y,z 18.415 z 56.01 w 19.605
(2.315) (2.956) (4.723) (8.478) (3.775) (3.535) (5.607) (3.054) (1.859)
1
Standard deviation.
a,b,c
Means within a row, which are not followed by a common superscript letter, are significantly different (P < 0.05).
within a column, which are not followed by a common subscript letter, are significantly different (P < 0.05).
w,x,y,z Means

Table 2––Effect of impregnation treatment (calcium lactate alone and chitosan + calcium lactate) on texture (firmness) of sliced
mushrooms impregnated at different vacuum pressures and different atmospheric restoration times during storage at 4 ◦ C.

Time 50 mm Hg 50 mm Hg 75 mm Hg 75 mm Hg 100 mm Hg 100 mm Hg 125 mm Hg 125 mm Hg

(days) Control –5 min –10 min –5 min –10 min –5 min –10 min –5 min –10 min
1 g/100 g calcium lactate solution (maximum force in newton)

0 a,b b a a a,b a,b a a b

w 30.297 y 34.61 y 25.125 y 29.106 y 30.443 y 32.313 z 27.525 y 24.84 z 36.147
1 (2.092) (0.573) (0.946) (0.965) (1.336) (2.370) (1.932) (0.47) (1.556)
4 a,b,c a,b,c,d b,c,d e d c,d d a a,b
w 28.257 x 29.74 z 31.87 z 40.717 y 33.65 y 33.408 y 33.57 y 26.226 x 27.773
(1.010) (3.109) (3.387) (2.368) (1.828) (1.374) (1.584) (1.740) (0.575)
9 e d b b,c a c a a b,c
y 47.79 z 40.503 y,z 26.97 y 31.71 x 19.666 y 33.066 x 14.943 x 15.03 y 31.478
(1.237) (1.752) (2.036) (1.528) (1.605) (1.720) (3.334) (1.58) (0.712)
12 b a a a a a
x 42.753 w 15.35 x 15.94 x 11.675 x 15.87 x 19.675
(2.409) (0.883) (0.806) (0.926) (1.256) (0.968) ∗∗∗ ∗∗∗ ∗∗∗
15 x 43.185
(2.315) ∗∗∗ ∗∗∗ ∗∗∗ ∗∗∗ ∗∗∗ ∗∗∗ ∗∗∗ ∗∗∗

1 g/100 g chitosan + 1 g/100 g calcium lactate solution (maximum force in newton)

0 w 30.297c y 31.972
c,d,e
y 25.52
a,b
x,y 36.182
e
x 29.9
b,c
y 22.47
a
y,z 32.426
c,d,e
y,z 30.787
c,d
y 35.17
d,e
1 (2.092) (2.012) (1.858) (2.173) (2.326) (0.980) (1.578) (1.622) (1.341)
4 a,b a,b,c b,c d c c c a a,b
w 28.257 x 21.02 y 28.615 y 38.713 x 31.583 z 44.81 y 30.56 z 37.846 y 30.8067
(1.010) (1.230) (2.525) (2.896) (2.323) (1.258) (1.654) (2.838) (2.731)
9 a,c c b b,c,d,e a,c a,d a,b,c a,b,c a,b,c
y 47.79 z 49.88 y 29.303 x 30.525 y 45.41 z 41.975 z 43.77 y 26.53 z 60.595
(1.237) (0.494) (0.241) (1.421) (0.895) (0.601) (7.254) (4.652) (3.486)
12 d b,c c,d c,d b,c a b,c a,b c,d
x 42.753 y 31.536 z 35.996 x,y 34.85 x 28.35 x 15.946 y,z 31.24 x,y 23.853 y 34.23
(2.409) (3.195) (2.296) (3.255) (1.456) (3.506) (1.103) (2.896) (5.624)
15 a b a,b a,b a,b a,b a,b
x 43.185 x 13.07 x,y 33.68 x 25.9 z 14.23 x 16.785 x 15.695
(2.315) ∗∗∗ (2.701) (3.422) (3.733) ∗∗∗ (4.101) (3.288) (5.395)
1
Standard deviation.
a,b,c
Means within a row, which are not followed by a common superscript letter, are significantly different (P < 0.05).
within a column, which are not followed by a common subscript letter, are significantly different (P < 0.05).
w,x,y,z Means

Vol. 79, Nr. 1, 2014 r Journal of Food Science E41

 Fresh-cut mushroom shelf-life extension . . .

Table 3–pH, color L∗ , and maximum force values for the con- Irradiation was carried out at room temperature using a 1.35 MeV
trol, impregnated, irradiated, and impregnated-irradiated sam- Van de Graaff e-beam accelerator. After irradiation, samples were
ples stored at 4 ◦ C for 15 d.
stored at 4 ◦ C up to 15 d for shelf life analysis.
Time Impregnated-
(days) Control Impregnated Irradiated Irradiated Quality parameters
pH Soluble solids. Soluble solids concentrations in the (a) im-
c a,b b,c a
pregnated, (b) irradiated, (c) impregnated-irradiated, and (d) con-
0 y 6.51 x 6.17 y 6.38 x 6.05
1 (0.08) (0.12) (0.08) (0.01)
trol samples were determined at room temperature using a hand-
4 y,x 6.30
a
x 6.18
a,b
y,x 6.30
a
x 6.15
a held refractometer (Brix 35HP, Reichert Analytical Instrument,
(0.08) (0.08) (0.11) (9.862) Inc., Buffalo, N.Y., U.S.A) and expressed in ◦ Brix scale. About 20
a a a a
9 x 6.18 x 6.07 x 6.17 y 6.23 to 25 g of mushrooms were placed in stomacher bags and squeezed
(0.12) (0.07) (0.10) (0.10) to yield around 10 g of juice. Measurements were carried out at
b a a a
x 6.18 x 6.11 x 6.20 x,y 6.19 room temperature and in triplicate for each treatment and control
12 (0.12) (0.06) (0.09) (7.796)
15 x 6.20
a
x 6.11
a
x 6.15
a
x 6.17
a group.
(0.03) (0.07) (0.03) (0.02) PH. The pH was measured using a digital pH meter (Cole
E: Food Engineering &

Parmer, pH 500 series, #59003-20, Singapore) calibrated with

Color L∗
Physical Properties

standard solutions, pH 4, 7, and 10 before the experiment. About

0 b a,b c a
z 70.701 z 70.142 z 73.333 w 68.503 20 to 25 g of mushrooms were placed in stomacher bags and
1 (2.501) (1.633) (2.185) (2.650)
c a b,c a,b
squeezed to yield around 10 g of juice. Measurements were carried
4 y 67.902 y 61.037 y 65.051 x,y,z 61.821 out at room temperature and in triplicate for each treatment and
(1.900) (4.298) (4.262) (6.275)
9 x 59.994
b
w 53.637
a
y 65.627
c
y 56.516
a,b control group.
(4.210) (2.599) (3.095) (5.109) Color. The color properties of the sliced mushrooms were de-
12 w,x 55.743
a,b
x 57.230
b
x 53.226
a
y 58.077
b termined using 20 pieces from the treated and control samples
(6.598) (2.454) (3.243) (3.384) at each sampling interval at room temperature using a Lab Scan
15 a b a,b c
w 52.631 w,x,y 57.216 x 55.186 z 63.525 XE colorimeter (Hunter Lab, Inc, Va., U.S.A.) calibrated with a
(3.293) (4.835) (2.823) (2.388)
Maximum force [N]
standard plate (Y = 94.00, x = 0.3578, y = 0.4567). Since our ob-
jective was to test whether the treatments maintained the whiteness
of the mushroom slices, only L∗ values (lightness) were reported.
0 b a b b
x 28.849 w 23.040 w 30.348 w 30.235
1 (5.571) (3.252) (4.178) (5.604)
a a a a The cap of the mushroom slices was placed in the aperture of the
4 x,y 34.095 y 37.328 w,x 35.985 w 33.563
(4.958) (6.322) (7.757) (9.862) colorimeter. New samples were used for each sampling interval
9 y 37.150
a,b
w,x,y 32.185
a
x,y 42.789
b
w 35.313
a,b (days 0, 4, 9, 12, and 15) to avoid microbial cross-contamination
(5.707) (8.353) (6.155) (10.574) of the slices.
b a c a,b
12 y 36.640 w,x 28.199 y,z 49.718 w 30.605 Texture. A shear test was applied with the Warner–Bratzler
(5.622) (6.168) (8.239) (7.796) probe at 1.0 mm/s using a Texture Analyzer (TA.XT2i, Texture
15 b,c a c a,b
y 44.210 x,y 31.781 z 51.575 w 39.670 Technologies Corp., Scardale, N.Y.). Texture was defined as the
(8.742) (6.538) (6.696) (7.378)
maximum force required to shear (cut) the samples. A preliminary
1
Standard
a,b,c
deviation. study showed that better results were obtained when the cap length
Means within a row, which are not followed by a common superscript letter, are
significantly different (P < 0.05). was reduced to 3 ± 0.2 cm by cutting the sides of the mushroom
w,x,y,z Means within a column, which are not followed by a common subscript letter, are slices and then combining 3 slices together. Thus, 3 slices (slice
significantly different (P < 0.05).

Figure 1–Mushroom slices treated under

different conditions (day 0 at 4 ◦ C.)

E42 Journal of Food Science r Vol. 79, Nr. 1, 2014

Fresh-cut mushroom shelf-life extension . . .
thickness approximately 6.5 ± 0.3 mm, total thickness approxi-
mately 19.5 mm) were used for the impregnation experiments. As
stated earlier, we had to adjust the thickness of the slices when
conducting the irradiation tests to ensure proper dose uniformity
within the slices. Therefore, samples consisted of 5 (instead of 3)
slices with total thickness of 19.5 mm. Ten replications were done
for each shear test at room temperature.
Microbiological analysis. Total aerobic plates, psy-
chrotrophic, and yeast and mold counts were determined on
days 0, 4, 9, 12, and 15 of storage. Under sterile conditions, 10
g of mushroom slices (cap and stem) from each treatment were
stomached inside a sterile stomacher bag, mixed with 90 mL
of 0.1 g/100 g buffered peptone water, and homogenized for
1 min; afterward, 10-fold dilutions were made in this diluent.
All counts were performed using petrifilms (3M yeast and mold
count plates, 3M aerobic plate count (APC), 3M microbiology,
St. Paul, Minn., U.S.A.). APCs were incubated at 37 ◦ C for 48 h;
psychrotrophic count plates at 4 ◦ C for 7 d; and yeast and mold
count plates were incubated at 20 ◦ C for 7 d. After incubation,
colonies were enumerated and results reported as log colony
forming units (CFU)/g of sample. The experiments were carried
out in triplicate.
Sensory evaluation. Thirty-five students, faculty, and staff
members at Texas A&M Univ. formed the consumer panel.
Evaluation of control, impregnated, irradiated, and impregnated-
irradiated slices was carried out under the same conditions on days
1, 9, and 15 of storage. The samples were placed into white plastic
plates labeled with 3 random digits and presented to the panelists
(Meilgaard and others 1998) who were asked to score the samples
based on odor, color, firmness, and overall quality using a 9-point
hedonic scale (a score of 1 represents “dislike extremely” and a
score of 9 represents “like extremely”). Scores higher than 5 were
considered acceptable.
Statistical analysis. Data analysis was performed using SPSS
software (version 20.0 for Windows 2011). Statistical differences
between variables were analyzed for significance by one-way anal-
ysis of variance using Tukey’s multiple range tests. Statistical sig-
nificance was expressed at the P < 0.05 level.
Results and Discussion
Preliminary study on VI
Color. By the end of the study (day 15), impregnation with
citric acid, calcium lactate alone, and chitosan solutions by VI did
not help maintain the lightness of mushroom slices, with L∗ val-
ues significantly (P < 0.05) lower than the control group (data
not shown). This effect was true for all combinations of vacuum
pressure and restoration time tested, demonstrating that these an-
tibrowning agents are ineffective and higher concentrations of
these agents may be required to achieve the desired browning in-
E: Food Engineering &
Physical Properties
hibition effect, which may induce flavor changes in the produce
(Harris and others 2007).
The 2 g/100 g ascorbic acid solution containing 1 g/100 g
calcium lactate applied at a 50 mm Hg for 5 min and 5 min
restoration time was the most effective VI treatment in terms of
color. L∗ values ranged from 60.56 on day 0 to 52.61 on day
15 compared to the control (72.91 and 48.6 on days 0 and 15,
respectively). This finding is not surprising because ascorbic acid is
the most effective antibrowning solution (Wang and others 2013)
because of the interaction of the antibrowning agent with calcium
and the mushroom’s tissue (Wang and others 2013). Perez-Cabrera
and others (2011) found similar results on a study with minimally
processed pears.
Texture. The maximum force to shear the control samples on
day 0 was around 30 N. On day 15, the force value was significantly
(P < 0.05) higher since the samples were very rubbery and hard to
shear. By day 15, the slices impregnated with ascorbic acid had the
most similar texture characteristics to the control group on day 0,
with differences (P < 0.05) due to the different conditions applied
during VI (applied vacuum pressures and atmospheric restoration
times) (Table 1).
Figure 2–Mushroom slices treated
under different conditions (day 15
at 4 ◦ C.)
Vol. 79, Nr. 1, 2014 r Journal of Food Science E43
 Fresh-cut mushroom shelf-life extension . . .

Table 4–Aerobic, psychrotrophics, and yeast and mold plate impregnated samples had moisture contents of 92.2% and 92.9%
counts for the control, impregnated, and impregnated- wet basis, respectively.
irradiated samples stored at 4 ◦ C for 15 d.
Next, we explored whether the above VI treatment combined
Aerobic plate count with irradiation at 1 kGy would help preserve the physicochemi-
Time Impregnated- cal, sensory, and microbiological quality of the mushroom slices.
(days) Control Impregnated Irradiated
0 c 4.891b
Combination Treatment: VI and Irradiation at 1 kGy
x 5.653 y N/D
1 (0.179) (0.098)
c b
Effect on quality attributes
4 y 7.164 x, y 4.980 N/D
(0.593) (0.458) PH. The pH of fresh mushrooms was approximately 6.5
9 y, z 7.817
c
x 6.560
b
N/D
(Table 3). The pH values of all the impregnated samples were
(0.404) (0.22) significantly (P < 0.05) lower than the nonimpregnated (control
b b
12 z 8.632 x, y, z 7.431 N/D and irradiated alone) samples, because of the acidic nature of ascor-
(0.169) (0.728) bic acid. On day 15, all samples had a pH of 6.2, still acceptable
15 c b
z 8.693 z 7.984 N/D
(0.206) (0.041)
for consumption (US FDA/CFSAN 2012).
Soluble solids (o Brix). All samples had ◦ Brix values between
E: Food Engineering &

Psychrotrophic plate count

Physical Properties

c
5.0 and 6.0 (data not shown) but no significant (P > 0.05) differ-
0 w 7.016 5.437b
x N/A
1 (0.134) ences were found among the samples.
(0.389)
4 7.684c b Color. By day 15, lightness values between the control and the
x x 5.331 N/D
(0.225) (0.159) treated samples were different (P < 0.05) (Table 3). On day 9, the
9 y 9.320
c
y 7.915
b
N/A impregnated samples had higher L∗ values than the control and
(0.059) (0.106) irradiated samples, probably due to the precipitation of calcium
12 c b
y, z 9.433 y 8.203 N/A in the cell walls, which may have increased the sample’s opacity.
(0.14) (0.616)
c b Furthermore, dark brown spots and dark staining occurred on the
15 z 9.774 y 8.815 N/A
(0.141) (0.379) surface of the controls. Figure 1 and 2 show the appearance of
Yeast and molds plate count the slices on days 0 and 15 of storage, respectively. Figure 2 shows
b the beneficial effect of the combined vacuum impregnation and
0 x 3.383 x 2.761b N/A
1 (0.686) (0.111) irradiation treatment in the color of the samples. Although ascorbic
4 x 3.620b x 3.043
b
N/D
acid is effective in preventing enzymatic browning once it oxidized
(0.573) (0.245) completely, darkening of samples occurred in impregnated samples
9 b b
x 4.412 x, y 3.960 N/D due to melanin formation. Irradiation seems to reduce browning
(0.206) (0.499) by slowing down enzyme oxidation (Niemira and Fan 2009).
12 b b 2.536a
x, y 5.625 x, y 4.621 x, y
(0.206) (0.927) (0.629)
Texture. Irradiation induced softening of the sliced mush-
15 y 4.458
b
y 4.985
b
y 3.370
a rooms (Table 3). This is a problem when exposing fruits and
(0.342) (0.091) (0.438) vegetables to ionizing radiation that causes depolymerization of
1
cellulose, hemicelluloses, starch, and pectin, which results in tis-
Standard deviation.
a,b,c
Means within a row, which are not followed by a common superscript letter, are sue softening (Niemira and Fan 2009; Moreira and Castell-Perez
significantly different (P < 0.05). 2012). VI also induced softening due to loss of structure. How-
w,x,y,z Means within a column, which are not followed by a common subscript letter, are
significantly different (P < 0.05). ever, application of the antibrowning solution (2 g/100 g ascorbic
Detection limit = 2.39 CFU/g. acid) containing 1 g/100 g calcium lactate maintained the firm-
N/A = No values observed, N/D = Under detection limit.
ness of the sliced mushrooms (approximately 35 N). Impregnated-
irradiated mushroom slices had more fresh-like texture than the
Impregnation with citric acid (Table 1) yielded samples with dif- irradiated slices, demonstrating the beneficial effect of VI with a
ferent texture characteristics. Similarly, calcium lactate alone was calcium-containing solution when samples are going to be irra-
not effective when applied by VI due to loss of quality in terms of diated. The controls and nonimpregnated samples suffered con-
structural losses (Table 2). The structural deformation was caused siderable structural loss (very soggy); hence, the blade could not
by the pressure changes during the VI process as observed by other shear the samples.
authors (Perez-Cabrera and others 2011). Increasing vacuum pres-
sure and vacuum time resulted in more structure deformation due Effect on Microbial Quality
to the applied pressures. Impregnation with chitosan (Table 2) Three sample groups were evaluated: (1) untreated control, (2)
produced sticky and rubbery samples by day 9 with force values impregnated, and (3) impregnated-irradiated.
lower (P < 0.05) than the controls by day 15. In brief, ascorbic
acid + calcium lactate impregnation at 50 mm Hg for 5 min and Aerobics
5 min restoration time effectively maintained the firmness of the Impregnation with ascorbic acid–calcium lactate solution was
sliced mushrooms stored at 4 ◦ C for 15 d. This VI treatment was effective in reducing the aerobics counts (P < 0.05) (Table 4). As
also effective in increasing the amount of physiologically active expected, the combined VI and irradiation treatment was effective
components in the mushroom composition with calcium increas- (P < 0.05) in reducing microbial growth (Kooropati and others
ing by 10 times and ascorbic acid by 150 times, compared to the 2004). The trend of microbial growth for the control and impreg-
USDA database values of 0.003 g/100 g and 0.0021 g/100 g, re- nated samples was similar, with an increase in counts with stor-
spectively (USDA 2011). After 5 min under 50 mm Hg and 5 min age time. Impregnation treatment alone reduced counts only by
restoration time, W1 = 22.96 ± 2.29 g, W2 = 27.19 ± 3.17, and 1-log compared to the combined treatment (under the detection
X = 18.42 ± 3.04%. Control and ascorbic acid–calcium lactate– limit).

E44 Journal of Food Science r Vol. 79, Nr. 1, 2014

Fresh-cut mushroom shelf-life extension . . .
Psychrotrophics
The organisms usually responsible for spoilage of mushrooms
are Gram-negative, psychrotrophic bacteria, particularly belong-
ing to the Pseudomonae family. These microorganisms are more
susceptible to irradiation than other types of spoilage bacteria
(Koorapati and others 2004). The combined VI and irradia-
tion treatment was very effective (P < 0.05) in reducing psy-
chrotrophic microbial growth, under the detection limit by day 9
(Table 4). Ascorbic acid impregnation reduced psychrotrophs by
1.5 logs, though this treatment alone is not sufficient to stop their
growth.
Yeast and molds
There was a significant (P < 0.05) difference in counts among
the control, impregnated, and impregnated-irradiated samples on
day 0 (Table 4). After day 12, there was a drastic increase in growth,
showing an approximately 2-log counts increase by day 15. This
finding suggests that the 1.0 kGy dose may be insufficient to
inactivate yeast and molds in sliced mushrooms. It is known that
yeasts are more resistant to irradiation than molds (da Silva Aquino
2012).
Figure 3–Sensory (A) color scores; (B) odor scores; (C) texture scores; and (D) overall quality scores for control, impregnated, irradiated, and impregnated-
irradiated samples stored at 4 ◦ C during 15 d. A score of 1 = dislike extremely, 5 = neither like nor dislike, and 9 = like extremely. A value above 5 is
considered acceptable.
Effect on sensory attributes
Sensory results indicated that the impregnated, irradiated,
and impregnated-irradiated samples were acceptable (P < 0.05)
(Figure 3). On day 1, there was no difference (P > 0.05) among
the color acceptability of the samples (Figure 3A). The control
samples were unacceptable to the panelists by day 15 of stor-
age (scores approximately 4.0), while the impregnated, irradiated,
and impregnated-irradiated samples were acceptable (scores ap-
proximately 6.0). This finding is supported by the objective color
measurements (Table 3). The odor scores of the control were
significantly (P < 0.05) different after day 1 (Figure 3B). Since
irradiation inhibited microbial growth, no odor changes were ob-
served in the irradiated and impregnated-irradiated samples with
scores > 6.0. Mushroom slices lost their original mushroom smell
with time, but treated samples did not exhibit unfavorable odor
E: Food Engineering &
changes.
Physical Properties
Throughout the evaluation period, the controls were less ac-
ceptable in terms of their texture attributes (Figure 3C). Although
the objective texture measurements showed clear changes in firm-
ness of the treated samples (Table 2), the panelists found all the
samples acceptable throughout storage (scores > 6.0).
The controls exhibited significant (P < 0.05) quality loss and
became unacceptable by day 15 of storage (Figure 3D). The
Vol. 79, Nr. 1, 2014 r Journal of Food Science E45
 Fresh-cut mushroom shelf-life extension . . .
impregnated, irradiated, and impregnated-irradiated samples had
consistently higher scores than the control group with the
impregnated-irradiated samples receiving slightly (though not sig-
nificant) higher scores ( >6.0).
Product appearance is the most appealing attribute to the con-
sumers. Since irradiation prevented microbial growth and re-
duced microbial-induced browning, irradiated and impregnated-
irradiated samples were consistently rated higher.
Conclusions
Although calcium is a texture enhancer for fresh-cut fruits and
vegetables, it was not effective in sliced mushrooms when applied
using VI and spoilage by fungi resulted in loss of quality. Irradi-
ation treatment alone caused softening of the sliced mushrooms;
however, impregnation with 1 g/100 g calcium lactate–ascorbic
E: Food Engineering &
acid solution helped to maintain mushroom firmness.
Physical Properties
Microbiological analysis demonstrated that e-beam irradiation
had an impact on reducing aerobic and psychrotrophic popula-
tions, whereas higher doses were required for inhibition of yeasts
and molds.
Sensory tests indicated consumers’ acceptance of the impreg-
nated, irradiated, and impregnated-irradiated mushrooms. Prod-
uct appearance was the most important quality attribute. Irradiated
and impregnated-irradiated samples had the highest overall sensory
scores because of reduced microbial-induced browning.
Results from this study demonstrated that e-beam irradiation
and VI of a solution containing 2 g/100 g of ascorbic acid and
1 g/100 g of calcium lactate can extend the shelf life of sliced
mushrooms.
Future work includes evaluation of the effect of impregnation-
irradiation on poyphenoloxidase activation to understand the
mechanism of browning inhibition; irradiation at doses higher than
1 kGy to reduce growth of yeasts and the effect on quality; eval-
uation of combination of ascorbic acid and citric acid solutions;
and optimization of impregnation solution and irradiation dose
combinations.
References
Akram K, Kwon J. 2010. Food irradiation for mushrooms: a review. J Korean Soc Appl Biol
Chem 53(3):257–65.
AOAC. 1990. Official methods of analysis of the association of office analytical chemists. 15th
ed. Washington, D.C.: Association of Official Analytical Chemists.
Beaulieu M, D’Aprano G, Lacroix M. 2002. Effect of dose rate of gamma irradiation on
biochemical quality and browning of mushrooms Agaricus bisporus. Radiat Phys Chem 63:311–
5.
Brennan M, Gromley TR. 1998. Extending the shelf life of fresh sliced mushrooms. Dublin.
Final Report Project ARMIS 4196.
Cliffe-Byrnes V, O’Beirne D. 2008. Effects of washing treatment on microbial and sensory
quality of modified atmosphere (MA) packaged fresh sliced mushroom (Agaricus bisporus).
Postharvest Biol Technol 48:283–94.
Da Silva Aquino, KA. 2012. Sterilization by gamma irradiation. In: Adrovic F, editor. Gamma
irradiation. Croatia: InTech Europe.
E46 Journal of Food Science r Vol. 79, Nr. 1, 2014
Duan Z, Xing Z, Shao Y, Zhao X. 2010. Effect of electron-beam irradiation on postharvest
quality and selected enzyme activities of the white button mushroom, Agaricus bisporus. J Agric
Food Chem 58:9617–21.
Fito P, Chiralt A, Betoret M, Gras MC, Martinez-Monzon J, Andres A, Vidal D. 2001. Vacuum
impregnation and osmotic dehydration in matrix engineering application in functional fresh
food development. J Food Eng 49(2–3):175–83.
Garcia E, Barret DM. 2002. Preservative treatments for fresh-cut fruits and vegetables. In:
Lamikanra O, editor. Fresh-cut fruits and vegetables: science, technology, and market. Chapter
9. Boca Raton, FL: CRC Press LLC, p 268–300.
Harris LJ, Yada S, Mitchman E. 2007. Publication 8229. University of California at Davis, p 20.
Hernandez-Munoz P, Almenar E, Ocio MJ, Gavara F. 2006. Effect of calcium dips and chitosan
coatings on postharvest life of strawberries. Postharvest Biol Technol 39:247–53.
Hironaka K, Kikuchi M, Koaze H, Sato T, Kojima M, Yaamamoto K, Yasuda K, Mori M, Tsuda
M. 2011. Ascorbic acid enrichment of whole potato tuber by vacuum-impregnation. Food
Chem 127:1114–8.
IAEA. 2002. Dosimetry for food irradiation. Technical reports series 409. Vienna, Austria: Intl
Atomic Energy Agency.
Koorapati A, Foley D, Pilling R, Prakash A. 2004. Electron-beam irradiation preserves the
quality of white button mushroom (Agaricus bisporus) slices. J Food Sci 69(1):25–9.
Lopez-Briones G, Varoquaux P, Chambroy Y, Bouquant J, Bureau G, Pascat B. 1992. Stor-
age of common mushroom under controlled atmospheres. Intl J Food Sci Technol 27:493–
505.
Mami Y, Gholamali P, Farhood Z, Mahmood G, Vahid S. 2013. Improvement of shelf-life
and postharvest quality of white button mushroom by 6o Co gamma-ray irradiation. Plant
Knowledge J 2(1):1–7.
Meilgaard M, Civille GV, Carr BT. 1998. Sensory evaluation techniques. 3rd ed. Boca Raton,
FL.: CRC Press. p 387.
Moreira RG, Castell-Perez ME. 2012. Irradiation applications in fruits and other fresh pro-
duce. In: Rodrigues S, Fernandes FAN, editors. Advances in fruit processing technologies
contemporary food engineering series. Chapter 7. Boca Raton, FL: CRC Press. p 203–17.
Niemira BA, Fan X. 2009. Irradiation enhances quality and safety of fresh and fresh-cut fruits
and vegetables. In: Niemira BA, Doona CJ, Feeherry FE, Gravani RB, editors. Micro-
bial safety of fresh produce. Chapter 10. Ames, IA: IFT Press, Wiley-Blackwell. p 191–
204.
Ortiz CF, Salvatori DM, Alzamora SM. 2003. Fortification of mushroom with calcium by
vacuum impregnation. Latin Am Appl Res 33:281–7.
Perez-Cabrera L, Chafer M, Chiralt A, Gonzalez-Martinez C. 2011. Effectiveness of antibrown-
ing agents applied by vacuum impregnation on minimally processed pear. LWT-Food Sci
Technol 44(10):2273–80.
Roy MK, Bahl N. 1984. Studies on gamma irradiation preservation of Agaricus bisporus. Mush-
room J 144:411–4.
Sapers GM, Miller LR, Miller FC, Cooke PH, Choi S. 1994. Enzymatic browning control in
minimally processed mushrooms. J Food Sci 59(5):1042–7.
Sapers GM, Miller RL, Pillizota V, Mattrazo AM. 2001. Antimicrobial treatments for minimally
processed cantaloupe melon. J Food Sci 66(2):345–9.
Singh P, Langowski HC, Wani AA, Saengerlaub S. 2010. Recent advances in extending the
shelf life of fresh Agaricus mushrooms: a review. J Sci Food Agric 90:1393–402.
Sommer I, Schwartz H, Solar S, Sontag G. 2010. Effect of gamma-irradiation on flavor 5’-
nucleotides, tyrosine, and phenylalanine in mushrooms (Agaricus bisporus). J Food Chem
123:171–4.
Teichmann A, Dutta PC, Staffas A, Jägerstad MJ. 2007. Sterol and vitamin D2 concentrations
in cultivated and wild grown mushrooms: effects of UV irradiation. LWT-Food Sci Technol
40:815–22.
USDA. 2011. U. S. Food and Drug Administration. National nutrient database for standard
reference. http://ndb.nal.usda.gov/ndb/foods/list. Accessed September 2013.
Vargas M, Chiralt A, Albors A, Gonzalez-Martinez C. 2009. Effect of chitosan-based edible coat-
ings applied by vacuum impregnation on quality preservation of fresh-cut carrot. Postharvest
Biol Technol 51:263–71.
Wang S, Lin T, Man G, Li H, Zhao L, Wu L, Liao X. 2013. Effects of anti-browning combinations
of ascorbic acid, citric acid, nitrogen and carbon dioxide on the quality of banana smoothies.
Food Bioprocess Technol 6: DOI: 10.1007/s11947-013-1107-7.
Wani AM, Hussain PR, Meena RS, Dar MD, Mir MA. 2009. Effect of gamma irradiation and
sulphitation treatments on keeping quality of white button mushroom Agaricus bisporus (J. Lge).
Intl J Food Sci Technol 44:967–73.
Yuk HG, Yoo MY, Yoon JW, Marshall DL, Oh DH. 2007. Effect of combined ozone and
organic acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on
enoki mushroom. Food Control 18:548–53.