Ultrasonics Sonochemistry 11 (2004) 379–384

www.elsevier.com/locate/ultsonch

The direct effect of ultrasound on the extraction of date

syrup and its micro-organisms
a,*
M.H. Entezari , S. Hagh Nazary b, M.H. Haddad Khodaparast b

a
Department of Chemistry, Ferdowsi University of Mashhad, 91775 Mashhad, Iran
b
Department of Food Science and Technology, Ferdowsi University of Mashhad, 91375 Mashhad, Iran
Received 22 April 2003; accepted 11 October 2003
Available online 19 November 2003

Abstract

In the date syrup industry, date fruits are mixed with a suitable amount of water at a temperature greater than 50
C for about 1
h. This condition is not sufficient for killing the micro-organisms present in the fruit. In addition, Overheating for a long time can
damages nutritious materials and also changes the final product’s color. Ultrasound was applied for improving the quantity and
quality of the extraction, and also to overcome the health problem. In this research, the following variables were examined: date
fruit/water ratio, ultrasonic intensity and temperature. The results showed that sonication under the proper conditions can lead to a
higher extraction in a shorter time with a better physical quality of the product. Most importantly, the sonication significantly
decreased the microbial count in comparison to the classical method. This study also confirmed the presence of anti-microbial
substances in date fruit, and that ultrasonic waves can accelerate their effects.
 2003 Elsevier B.V. All rights reserved.

Keywords: Ultrasound; Extraction; Date syrup; Micro-organism; Anti-microbial

1. Introduction used for the date syrup production with economical

The total amount of date fruit production in the world
with main producer countries in 1997 is shown in Fig. 1.
The highest amount of date fruit production belongs to
Iran with 20% of the total world production [1]. Kabkab
date is a type of Iranian date which has a high amount of
wastes; this is due to its high moisture and tenacity. Only
35% of this date is used as first class, with the remainder
used for second and third degree date and wastes [2].
Therefore, this research was focused on this type of date.
Date fruit as a nutritive product contains sugar sub-
stances about (70.6–76.3%), proteins (1.9–3%), fat (0.2–
2.8%), minerals (1.3%) and vitamins [3].
In food technology, date syrup as the main and
general byproduct of date is used for foodstuffs such as
jams, marmalades, concentrated beverages, chocolates,
ice cream, confectioneries, honey, and etc. As the
Kabkab date has a high amount of wastes, it can be
*
Corresponding author. Fax: +98-511-843803.
E-mail address: moh_entezari@yahoo.com (M.H. Entezari).
advantageous.
In date’s syrup industry the fruit are mixed with water
and heated for around 1 h at 50
C, when the main
components, sugars, are extracted. This method destroys
some nutritive components and darkens the product’s
color. The temperature used is not high enough to kill the
micro-organisms naturally present in syrup and there-
fore this product raises some health problems.
Acoustic cavitation is the formation, growth and
collapse of bubbles in liquids induced by ultrasonic
waves. A number of papers have dealt with the ultra-
sonically assisted extraction of different materials [4–9].
High efficiency and shortening of extraction time are the
main positive effects of such extractions. The most
important proposed mechanism for these behaviors is
the intensification of mass transfer and easier access of
the solvent to the content of the cell. In addition, the
collapse of cavity near the cell wall can lead to cell
disruption and releases the cell contents into the
extraction medium [10–12]. Ultrasound can also inacti-
vate a range of micro-organisms. An investigation

1350-4177/$ - see front matter  2003 Elsevier B.V. All rights reserved.
doi:10.1016/j.ultsonch.2003.10.005
380 M.H. Entezari et al. / Ultrasonics Sonochemistry 11 (2004) 379–384
Fig. 1. Total amount of date fruit production in the world with main
producer countries in 1997.
explored the effects of ultrasonic waves on bacteria,
fungi, and viruses [13]. Ultrasound in the range of 20–
100 kHz frequency is mainly used in industrial processes
[14] and that those with higher frequency are mainly
used for detection and analyses [15].
The main objective of this work is to focus on the
comparison of conventional and ultrasonically assisted
extraction of Kabkab date syrup. In this study also the
effect of ultrasound on the micro-organisms under dif-
ferent applied conditions are discussed.
2. Experimental
2.1. Materials and Instruments
• Date fruits: Kabkab dates with an average contain of
sugar of 65% (analyzed by Lane–Eynons Method)
was used in all experiments. To prevent useful com-
ponent losses it was stored in a cold room at 10
C.
• Ultrasound device: ÔMisonix’ model XL-2020, 20
kHz, 500 W, horn type machine with 12.7 mm tip
diameter.
• Accurate refractometer was used for determination
of soluble solids with main component of sugar in
date’s syrup.
• Mother microbial culture: It was prepared by incu-
bating crushed date fruit plus water at 30
C in open
vessel for 24 h. Then nutrient broth (DIFCO) plus 1%
glucose medium enriched it for 24 h at 30
C.
2.2. Applied methods
The dates were sliced to pieces with the approximate
dimensions of 1 cm · 1 cm · 0.3 cm. Then the material
obtained was used for four extraction methods. The
water/fruits ratio was 3/1 (w/w) and 9/1 (w/w). In a
typical experiment (9/1), in an Erlenmeyer flask (100 ml)
5 g of date pieces and 45 g of water were mixed and
extracted using method described below.
First method: magnetic stirring of mixture.
Second method: mixture water/fruits let to stand
without mixing.
Third method: Sonication, 20 kHz, 25% from total
power of the ultrasonic device.
Fourth method: Sonication, 20 kHz, 10% from total
power of the ultrasonic device.
All experiments were performed at two temperatures:
15 and 35
C.
In first method, the experiments were carried out in
an Erlenmeyer flask (100 ml) using a magnetic stirrer at
rotation speed of 420 rpm. In ultrasonically method, the
samples were extracted by continuous ultrasonic waves
with 20 kHz frequency in two levels of intensity. The
volume and the kind of vessel was the same as classical
method and the distance from probe tip to vessel’s
bottom was set to 1 cm.
All experiments were performed in sterile conditions
for 90 min and samples were withdrawn at 10 min
intervals. Sampling for microbial cultures was carried
out at the start and end of extraction. Then samples
inoculated in two solid culture mediums consist of
nutrient agar (DIFCO) with 2% glucose and malt ex-
tract agar (MERK), for aerobic mesophyle micro-
organisms and fungi growth respectively.
Abbreviation for samples:
S1–S4: Control samples, stirring
S5–S8: Control samples, silent
U1–U4: Sonication, power setup at 25%
U5–U8: Sonication, power setup at 10%
The conditions for the samples are as follows:
S1, U1, S5, U5: the water/fruits ratio ¼ 3/1, the tem-
perature ¼ 15
C
S2, U2, S6, U6: the water/fruits ratio ¼ 9/1, the tem-
perature ¼ 15
C
S3, U3, S7, U7: the water/fruits ratio ¼ 3/1, the tem-
perature ¼ 35
C
S4, U4, S8, U8: the water/fruits ratio ¼ 9/1, the tem-
perature ¼ 35
C
The produced syrup was filtered by filtering cloth and
then centrifuged at 5000 rpm for 15 min.
3. Results and discussions
3.1. Classical extraction
These experiments were conducted by mechanical
stirring. Fig. 2 demonstrates the extraction by Brix/total
mass of the date fruit against of time for the silent and
mixing extractions. It is clear that the extraction rate is
 M.H. Entezari et al. / Ultrasonics Sonochemistry 11 (2004) 379–384
Fig. 2. Brix/total mass variation among 90 min of extraction in Control
samples (Mixe ¼ S and silent ¼ s): S1; S2;
S4; s5; s6; s7; s8.
higher by mixing as compared to the silent one. The
reason for the low extraction rate in the silent method
can be due to the formation of saturated layer of ex-
tracted materials around the date fruit and this phe-
nomenon can almost stop the extraction of materials.
The mechanical stirring removes this layer from around
the pieces of date fruit and then the pieces contact with
new solvent, resulting in a higher extraction rate. This
figure also shows the extent of increasing from silent to
mixing which is different from this system to other sys-
tems. The extraction rate is affected by changing the
temperature and the ratio of water to date fruit for both
classical methods. The classical extraction is based on
the diffusion process and this physical property is in-
creased by increasing the temperature. Therefore, in
both classical methods better extraction were achieved
at higher temperature. The water/fruits ratio also af-
fected differently the extraction rate in both classical
methods. Higher extraction efficiency was achieved at
lower ratio in silent method and at higher ratio in
mixing method. This difference can also be explained by
the more contact of solid phase (date) to liquid phase
(water) in the mixing method at higher ratio and in the
silent case the more important is the higher gradient
of concentration which achieved at lower ratio.
3.2. Ultrasonic extraction
These experiments were carried out under different
variables such as intensity, the ratio of solid to liquid
phase, and temperature. Fig. 3 demonstrates the effect of
these variables on the extraction rate. Higher intensity
was more effective than lower one on the extraction rate.
This could be explained that bubbles collapse more
violently at higher intensity than lower one, leading to
more extraction.
The ratio had different effects, with higher intensity
the ratio of 9/1 was better than 3/1, but this behavior
was reversed with lower intensity. This could be ex-
381
Fig. 3. Brix/total mass variation among 90 min of extraction in Soni-
cated samples: U1; U2; U3; U4; U5;
S3;
U6; U7; U8.
plained by the combination of ultrasonic extraction and
thermal extraction. In higher ratio of water to fruit date
(9/1); the ultrasonic effect is more pronounced than
thermal effect. This is due to the better performance of
acoustic cavitation in dilute solution (9/1) than viscous
one (3/1).
Ultrasonic cavitation is more effective at lower tem-
peratures; therefore the optimum condition for the
extraction was achieved at higher intensity, higher ratio,
and lower temperature. In ratio of 3/1, due to the high
viscosity of the solution the effect of ultrasound on
extraction was negligible and the thermal extraction was
predominant and this leads to more extraction at higher
temperature than lower one.
3.3. Comparison of ultrasonic and classical extraction
The sample U2 was the best from point of view of the
rate, the yield, and the quality of extracted materials. In
this sample the ultrasonic waves at high intensity
introduced directly to the solution with the water/fruits
ratio of 9/1 at 15
C. Fig. 4 compares the samples U2
and S2 of which the latter one is the same as the first one
Fig. 4. Comparison of ultrasonic extraction with classical method
under the same conditions: S2; U2.
382 M.H. Entezari et al. / Ultrasonics Sonochemistry 11 (2004) 379–384
except it was carried out without ultrasound. As it is
shown, the Brix/total mass for sample U2 at 10 and 20
min is equal to 40 and 90 min of the sample S2,
respectively. This figure clearly confirms that the ultra-
sonic extraction is more effective than classical extrac-
tion in terms of the yield and the rate. The more
important point is the higher initial slope in ultrasonic
extraction, which leads to the completion in the shorter
time (about 20 min). In this period of time, the taste of
sonicated sample was sweeter than conventional one.
This confirms that the ultrasonic extraction can be car-
ried out not only in the shorter time but also in the lower
temperature. Both of these parameters are very impor-
tant in food industry.
With regard to the products’ color, there was also
difference between classical and ultrasonic extraction.
Control samples had a red color which was changed to a
stronger color at higher temperature. Samples under
sonication had a cream color at lower temperature but
at higher temperature the color changed to brown.
From point of view of the color, sonicated sample is
better than control samples. In industry, the color of
the control sample is changed to cream by the addition
of chemicals.
4. Micro-organism and ultrasound
4.1. Total aerobic mesophyl count
Fig. 5 demonstrates the percent of total microbial
count for different samples. These samples behaved
differently and can be divided to three groups. In control
samples as a first group (S1–S4), a decrease in the total
microbial count is observed. This reduction could be
explained by the presence of anti-microbial components
in the date fruit. The anti-microbial effect is varied from
12% to 52% on these samples. The different effects can
be related to the concentration of antimicrobial com-
Fig. 5. Reduction of total microbial count under classical method and
sonication: TC 90.
ponents which is higher at 3/1 ratio than 9/1 one. The
experimental results also show that the lower tempera-
ture is more effective (S1) for reduction than higher one
(S3).
In the second group (U1–U4), ultrasonic waves with
high intensity removed more micro-organisms. A com-
bination of physical and chemical mechanisms which
occur during acoustic cavitation might be responsible
for the reducing of micro-organisms. The chemical
mechanism for this reduction is attributed to attack by
free radicals, especially hydroxyl and also due to the
hydrogen peroxide produced. The physical mechanism
can be attributed to the disruption of cell membrane
arising from bubble implosion. Ultrasound can also
facilitate the disaggolomeration of micro-organism
clusters in solution which lead to increase the killing of
them by anti-microbial compounds from date syrup.
In the third group (U5–U8), ultrasonic waves with
low intensity reduced micro-organism with a lower rate.
This is due to the cavitation process producing milder
conditions. Between the sonication samples, U1 was the
best for destroying the micro-organisms. Under this
condition 85% of the total microbial count was de-
stroyed after 90 min. Between the classical samples, S1
was the best and 52% of micro-organisms have de-
stroyed. Comparison of these two results clearly shows
that under sonication 33% more micro-organisms were
destroyed than with the classical method.
For illustrating the microbial view before and after
sonication, inoculated date syrup sample was spread on
the lam surface and then stained by gram manner. Fig. 6
shows the mixed culture of micro-organisms of the
mixture which contains date fruit and water before
sonication, i.e. at the starting point of extraction. Fig. 7
Fig. 6. The picture of mixed culture of micro-organisms before soni-
cation of the date syrup.
 M.H. Entezari et al. / Ultrasonics Sonochemistry 11 (2004) 379–384
Fig. 7. The picture of mixed culture of micro-organisms after sonica-
tion of the date syrup.
demonstrates the mixed culture of micro-organisms
after sonication of the date syrup at the end of extrac-
tion. The comparison of these two Figs. 6 and 7 clarified
that the ultrasounic wave disperses and damages the
microbial cells and also discharges the cells’ contents.
This can be explained by the physical and chemical
effects of ultrasound. As the syrup is a nutrient for mi-
cro-organisms, some additives to prevent microbes
growing might be necessary.
4.2. Fungi count
Fig. 8 demonstrates the percent of fungi count for
different samples. As for Fig. 5, there are three different
groups. The most important point is that the ultrasonic
waves with low intensity not only did not reduce the
fungi count, but also enhanced the growth of their cells
and increases fungi count. This could be because the low
Fig. 8. Reduction of fungi count under classical method and sonica-
tion: Fungi 90.
383
intensity cannot destroy the membrane of the cell, but
can facilitate the increase of mass transfer between in-
side and outside the cell. In other words, the ultrasonic
wave increases the metabolism of the cells. From point
of view of reduction of fungi count, sample U2 (high
intensity) was the best and under this condition 69% of
the fungi count has destroyed. Comparison of U2 and
S2 also shows an increase of 28% on the fungi destruc-
tion under sonication. In contrast, ultrasonic waves
with low intensity increases the fungi count to about
11–13.4%.
5. Conclusion
From the point of view of extraction, the ultrasonic
waves with high intensity and low temperature were
better for date fruit; condition U2 (at 25% of power, at
15
C, and at water/fruits ratio of 9/1) was selected as
the best for the extraction. The important result is that
the time of extraction is shorter under sonication, a
very important point in food industry. The time of
extraction has a key role on the quality of extracted
materials such as the color and the nutritious material
of products. This investigation also confirms the pres-
ence of anti-microbial substances in date fruit, and that
ultrasonic waves enhance their effectiveness. With re-
gard to micro-organisms, the ultrasonic waves with
high intensity reduced the total microbial counts. An-
other interesting point is that the effect of ultrasonic
waves with low intensity on the fungi count is to in-
crease its number.
References
[1] Date fruit position in Iran and world illustrated by figures,
Agriculture Ministry of Iran, 2001.
[2] A. Saifcordi, et al., To survey of converting process of Bushehr’s
Kabkab, Sharif University of Tehran, 1992.
[3] M. Vosooughy, I. Alamzadeh, To survey of preparation of new
product from date fruit, Agriculture Ministry of Iran, 1993.
[4] Z. Hromadkova, A. Ebringerova, P. Valachovic, Ultrasound-
assisted extraction of water-soluble polysaccharides from the roots
of valerian, Ultrason. Sonochem. 9 (2002) 37–44.
[5] M. Toma, M. Vinatoru, L. Paniwnyk, T.J. Mason, Investigation
of the effects of ultrasound on vegetal tissues during solvent
extraction, Ultrason. Sonochem. 8 (2001) 137–142.
[6] M. Vinatoru, An overview of the ultrasonically assisted extraction
of bioactive principles from herbs, Ultrason. Sonochem. 8 (2001)
303–313.
[7] I. Panchev et al., Improving pectin technology, extraction using
ultrasonic treatment, Int. J. Food Sci. Technol. 23 (1988) 337–341.
[8] L.L. Wang, Ultrasound extraction of proteins from autoclaved
soy been flakes, J. Food Sci. 40 (1975) 549–551.
[9] H. Fukase et al., Effect of ultrasonic in soy been Protein
extraction, J. Appl. Phys. 33 (1994) 3088–3090.
[10] T.J. Mason, Y. Zhao, Enhanced extraction of tea solids, Ultra-
sonics 32 (1994) 375–377.
384 M.H. Entezari et al. / Ultrasonics Sonochemistry 11 (2004) 379–384

[11] A.E. De Maggio, J.A. Lolt, J. Pharm. Sci. 53 (1964) 945.
[12] K.J. Moulton, L.C. Wang, J. Food Sci. 47 (1982) 1127.
[13] G. Scherba, R.M. Weigel, W.D. O’Brien Jr., Quantitative
assesment of the germicidal efficacy of ultrasonic energy, Appl.
Environ. Microbiol. 57 (1991) 2079–2084.
[14] T.J. Mason, L. Paniwnyk, The uses of ultrasound in food
processing, in: Advances in Sonochemistry, vol. 4, JAI Press
Inc., 1996, pp. 177–203.
[15] J.D. Mc Clements, Ultrasonic characterization of foods, drinks,
Food Sci. Nutrition 37 (1997) 1–46.