You are on page 1of 5

Available online at www.sciencedirect.

com

Polymer Degradation and Stability 93 (2008) 268e272


www.elsevier.com/locate/polydegstab

Effects of ultrasonic extraction on the physical and chemical properties


of polysaccharides from longan fruit pericarp
Bao Yang a, Yueming Jiang a,*, Mouming Zhao b, John Shi c, Lingzhao Wang d
b

a
South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, Guangdong 510650, PR China
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, Guangdong 510640, PR China
c
Food Research Center, Agriculture and Agri-Food Canada, Guelph ON N1G 5C9, Canada
d
School of Ocean, Huaihai Institute of Technology, Lianyungang 222005, PR China

Received 13 July 2007; received in revised form 29 August 2007; accepted 13 September 2007
Available online 2 October 2007

Abstract
An ultrasonic technique was employed to extract polysaccharides from longan fruit pericarp (PLFP). Effects of ultrasonic power, time and
temperature on the extraction of PLFP were examined. Different effects of ultrasonic time were observed at two different ultrasonic power of 120
and 300 W. A higher recovery rate of PLFP at an ultrasonic power of 300 W was obtained as compared with 120 W. The recovery rate of PLFP
was slightly increased by elevating the ultrasonic temperature up to 60  C. The highest recovery rate of PLFP was achieved at 120 W and 70  C
for 20 min. Furthermore, PLFP I and PLFP IIeIV were prepared by hot-water extraction and ultrasonic extraction, respectively, and then used
for the analyses of physical and chemical properties. Analysis by differential scanning calorimetry showed that the onset temperature, peak temperature, conclusion temperature and melting enthalpy (DH ) of PLFP by hot-water extraction were lower than those by ultrasonic extraction.
These results suggested that rearrangement of PLFP microstructure could occur and development of a higher proportion of crystalline regions
might be induced by the ultrasonic treatments. The highest DH (8.02 J/g) and two endothermic peaks were observed in the thermogram of PLFP
II. Scanning electron micrographs revealed more aggregated particles in PLFP III and IV compared with PLFP I and II. However, no apparent
differences were found from the spectra of these four PLFP samples at a range of 195e550 nm, which indicated that ultrasonic treatment might
not cause significant chemical modification of groups in the PLFP chain.
2007 Elsevier Ltd. All rights reserved.
Keywords: Longan; Polysaccharide; Ultrasonic extraction; Recovery; Differential scanning calorimetry

1. Introduction
Polysaccharides play an important role in the growth and
development of living organisms. They have been widely studied in recent years due to their unique biological, chemical and
physical properties [1]. Thus, polysaccharides as important
components of therapeutic drugs and skin care products could
be used in modern medicine [2,3].
Recently, ultrasonic extraction was tested as an alternative
method for isolating polysaccharides from plant materials [4].
It is reported that extraction efficiency is enhanced greatly by
* Corresponding author. Tel.: 86 20 37252525; fax: 86 20 37252831.
E-mail address: ymjiang@scbg.ac.cn (Y. Jiang).
0141-3910/$ - see front matter 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.polymdegradstab.2007.09.007

ultrasonic treatments. The ability of ultrasonic treatment to


improve the recovery of polysaccharides is mainly attributed
to its facilitation of mass transfer between immiscible phases
through agitation, especially at low frequency [5]. The most
important mechanical effects of ultrasonic treatments are microjetting and microstreaming [6,7]. The former is a consequence of cavitation at or near large solid surfaces, resulting
in bubble collapse in an asymmetric way. The latter is mainly
a consequence of cavitation in the presence of suspended powder. Enhancement of plant material hydration and fragmentation are the primary benefits of ultrasonication that result in
a high recovery of polysaccharides. However, ultrasonic treatment can have a detrimental effect on polysaccharides to
some extent, depending on the applied power and operating

B. Yang et al. / Polymer Degradation and Stability 93 (2008) 268e272

269

parameters [8], and can lead to changes in structure and other


properties of polysaccharides [9]. It is well known that the
bioactivity and functional properties of polysaccharides are related to the structure and molecular weight [10]. Therefore,
a greater understanding of the effects of ultrasonic treatments
on the physical and chemical nature of polysaccharides is
required.
Longan (Dimocarpus longan Lour.) is an exotic fruit in
Southeast Asia, especially in China [11]. Our previous work
indicated that longan fruit pericarp tissues were comprised
of extensive quantities of polysaccharides. In the present
study, an ultrasonic technique was employed to extract
PLFP. Furthermore, the recovery of PLFP extraction was
evaluated, while the thermodynamic and morphological properties as well as the UVevis spectra of PLFP samples prepared both by hot-water extraction and ultrasonic extraction
were determined using differential scanning calorimetry
(DSC), scanning electron microscopy and UVevis spectrophotometry, respectively.

China, 40 kHz). Four grams of pericarp powders were extracted in 100 ml of distilled water in a 150-ml conical flask.
The flask was held in an ultrasonic cleaner with water and exposed to 5, 10, 20, 30 or 40 min of ultrasonication at 25, 40,
55 or 70  C and 120 or 300 W of ultrasonic power. The subsequent filtration and purification of polysaccharides were
conducted according to the hot-water extraction method as
described above. PLFP I was extracted by hot-water extraction and PLFP IIeIV were extracted by ultrasonic treatments
using the lowest or highest ultrasonic power and temperature.
Their preparation conditions are as follows: PLFP I: 20 min
and 25  C; PLFP II: 120 W, 20 min and 25  C; PLFP III:
120 W, 20 min and 70  C and PLFP IV: 300 W, 20 min and
25  C.
The contents of polysaccharides were determined by the
phenolesulfuric acid method [14]. Glucose was used to
make a standard curve. The recovery of PLFP was expressed
as milligrams of glucose equivalents (GE) per gram of pericarp tissues on dry weight (DW) basis.

2. Experimental

2.4. Thermodynamic analysis

2.1. Plant materials

The thermodynamic characteristics of PLFP samples were


determined using a Diamond DSC (PerkineElmer, Massachusetts, USA). PLFP I, II, III or IV was diluted with 5-volumes
of distilled water. The mixture (10 mg) was weighed and then
placed in an aluminium pan. The pan was sealed hermetically
and heated from 30 to 160  C at a rate of 10  C/min. The onset
temperature (To), peak temperature (Tp), conclusion temperature (Tc) and melting enthalpy (DH ) were recorded.

Fresh longan (D. longan Lour. cv. Shixia) fruits at the commercially mature stage were purchased from a commercial
market in Guangzhou, China. Fruits were selected for uniformity of shape and colour.
2.2. Chemicals
Glucose, phenol and sulfuric acid were obtained from the
Guangzhou Reagent Co. (Guangzhou, China). All other chemicals used were of analytical grade.
2.3. Extraction and quantification of PLFP
PLFPs were extracted by the hot-water isolation method of
Yang et al. [12]. The longan fruit pericarp tissues were airdried and crushed using a mortar and pestle. The crushed pericarp tissues were passed through a 60-mesh stainless steel
sieve to obtain the pericarp powder. Four grams of pericarp
powders were extracted for 5, 10, 20, 30 or 40 min in
100 ml of distilled water in a 150-ml conical flask submerged
in a water bath at 25, 40, 55 or 70  C, and then filtered through
a Whatman No. 1 paper (Whatman Co., Shanghai, China). The
filtrate was concentrated to 25 ml using a rotary evaporator
(BC-R203, Shanghai Biochemical Equipment Co., Shanghai,
China) at 65  C under low pressure. The proteins in the extract
were removed using the Sevag reagent [13]. After removal of
the Sevag reagent, 100 ml of anhydrous ethanol was added and
the mixture was maintained overnight at 4  C to precipitate
polysaccharides. PLFP was then obtained by centrifugation
at 3860g for 15 min.
Extraction by ultrasonication was performed using an ultrasonic cleaner (SB-5200DTD, Xinzhi Biotech Co., Ningbo,

2.5. Morphological properties


PLFP samples were applied to copper stubs using doublesided adhesive tapes and coated with gold powders. Mounted
samples were observed using a JSM-6360LV scanning electron microscope (JEOL, Tokyo, Japan) at an accelerating
potential of 15 kV.
2.6. UVevis spectrophotometric analysis
The UVevis spectrophotometric analysis was conducted
according to the method of Zhao et al. [15] with a minor modification. Each PLFP sample (1 mg) was dissolved in 10 ml of
distilled water. The absorbance of each sample solution was
read over the range of 195e550 nm, using a UV-2102 PC
UVevisible spectrophotometer (Unico, Shanghai, China).
2.7. Statistical analyses
Data were expressed as means  standard errors (SE) of
three replicated determinations. Analysis of variance (ANOVA) was applied for determining significant difference at
P < 0.05 using OriginPro Version 7.5 software (OriginLab
Corporation, Northampton, USA).

B. Yang et al. / Polymer Degradation and Stability 93 (2008) 268e272

270

3. Results and discussion


3.1. Effects of ultrasonic power, time and temperature on
the recovery of PLFP
The effect of ultrasonic time on the recovery of PLFP is
shown in Fig. 1. Ultrasonic extraction was more efficient in
obtaining PLFP than hot-water extraction. Recovery rate of
PLFP by hot-water extraction increased gradually over the
40-min experiment. In contrast, the recovery rate of PLFP
by ultrasonication at 120 W reached a maximum rate after
just a 20-min extraction. Longer treatment time did not improve the rates of polysaccharide isolation. The recovery rates
of PLFP at 300 W and 25  C for 10, 20, 30 or 40 min were significantly (P < 0.01) lower than those at 120 W and 25  C for
the same ultrasonic treatment time. The highest recovery rate
(4.99 mg GE/g DW) of PLFP was obtained at 120 W for
20 min when 25  C was used.
The effect of ultrasonication temperature on the recovery
rate of PLFP is presented in Fig. 2. The recovery rate of
PLFP was significantly (P < 0.01) higher via ultrasonication
at 120 W than at 300 W and those obtained by hot-water extraction. Using an ultrasonic power of 120 W or hot-water extraction, the recovery rate of PLFP slightly increased with
elevating ultrasonic temperature. However, using an ultrasonic
power of 300 W, it increased with elevating ultrasonic temperature up to 60  C and then decreased. The highest recovery
rate (5.49 mg GE/g DW) of PLFP was achieved at 120 W
and 70  C when a 20-min ultrasonic treatment time was used.
The mechanism of ultrasonic extraction involves two physical processes: the dissolution of the extracted polysaccharides
near the particle surface (rinsing) and the diffusion of polysaccharides to the liquid extract (slow extraction) [16]. Increasing
the ultrasonic time could enhance these two activities, resulting
in a high extraction recovery rate, which might account for the
increase observed in the recovery rate of PLFP at 120 W as
the treatment time was extended from 5 to 20 min. The lower recovery rate of PLFP at 300 W may relate to degradation effects
of polysaccharides by ultrasonic wave. It was reported that
higher ultrasonic power resulted in strong degradation effects
[17,18]. At high ultrasonic temperature, the liquid viscosity

Recovery of PLFP
(mg GE/g DW)

5.5
5
4.5
4
3.5
3
20

30

40

50

60

70

Ultrasonication temperature (C)


Fig. 2. Effect of ultrasonication temperature on the recovery rate of PLFP. The
ultrasonic time used was 20 min. -, :: ultrasonic extraction at 120 and
300 W, respectively. A: hot-water extraction. Data are presented as means 
SE of three replicated determinations.

and density decreased resulting in a more rapid mass transfer


[19]. Furthermore, high ultrasonic temperature can lead to an increase in the number of cavitations within these tissues and surface contact areas [20]. Thus, application of high ultrasonic
temperature resulted in enhanced extraction efficiency.
3.2. Thermodynamic properties
The To, Tp, Tc and DH of PLFP I were determined to be
125, 131, 140  C and 2.5 J/g, respectively. They were lower
than those of PLFP IIeIV, which indicated that ultrasonic
treatment caused modifications in the microstructure of
PLFP. The increased Tc of ultrasonic-treated PLFP implied
that more extended junction zones of PLFP occurred [21].
The apparent DH calculated from the DSC curves indicated
the required energy for disrupting hydrogen bonds within the
junction zones [22]. A stronger carbohydrateewater interaction and better-organized microstructure can lead to a higher
DH [23]. PLFP II had the highest DH (8.02 J/g), followed
by PLFP III (4.8 J/g) and PLFP IV (3.3 J/g). These differences
may be due to a moderate ultrasonic treatment resulting in

Recovery of PLFP
(mg GE/g DW)

6
5
4
3
2
0

10

20

30

40

Ultrasonication time (min)


Fig. 1. Effect of ultrasonication time on the recovery rate of PLFP. The ultrasonication temperature used was 25  C. -, :: extraction at 120 and 300 W,
respectively. A: hot-water extraction. Data are presented as means  SE of
three replicated determinations.

Fig. 3. DSC thermograms of PLFP IeIV. 1: PLFP I, 2: PLFP II, 3: PLFP III
and 4: PLFP IV.

B. Yang et al. / Polymer Degradation and Stability 93 (2008) 268e272

a better-organized microstructure of PLFP. Unlike the other


three samples, two endothermic peaks at 134 and 138  C
were observed in the thermogram of PLFP II (Fig. 3). The
sharp rising curve at the end of the thermogram was due to
the transition of free water into gaseous state. According to
the endothermic transition of PLFP, the physical nature of
the polysaccharide might be a partially crystalline polymer
consisting of crystalline and amorphous regions [24]. As documented by some researchers [25], the crystal perfection phenomenon was related to an increase in the melting
temperature and enthalpy value. This study suggested that ultrasonic treatment might lead to a higher proportion of crystalline regions in PLFP but it requires to be determined further,
for example, by X-ray diffraction.
3.3. Morphological properties
The surfaces of gold-coated PLFP IeIV were imaged by
a scanning electron microscope. The micrographs at 6500fold magnification are presented in Fig. 4. The image of
PLFP I contains clear and small particles distributed discretely
(Fig. 4A). After ultrasonic treatment at 120 W and 25  C for
20 min, smaller particles developed (Fig. 4B). In Fig. 4C
and D, the shape of some particles was unclear while many aggregated particles appeared, which might be due to the degradation and aggregation effects of polysaccharides by
ultrasonic treatment [26,27]. Szu et al. [28] reported that

271

ultrasonic irradiation for depolymerisation was effective in


preparation of homogeneous and low molecular weight polysaccharides. It is generally accepted that ultrasonic irradiation
can depolymerise the polysaccharide due to the cavity effect
while in the vicinity of a collapsing cavity rapid motion of solvent generates shearing forces that break chemical bonds in
the polymer [28]. In this study, the increase in ultrasonic
power from 120 to 300 W at 25  C resulted in more aggregates
of PLFP than the elevation of sonication temperature from 25
to 70  C when an ultrasonic power of 120 W was used. The
result suggested that ultrasonic power was an effective factor
for modifying the physical property of PLFP. The change in
the morphological characteristic among these four PLFP samples might contribute to the modification in the molecular
weight and other physical and chemical properties.
In the continual search for improved degrading or re-associating pathways, recent emphasis has been given to ultrasound [29]. It has been pointed out that the sonochemical
method is potentially useful in carbohydrate chemistry. Firstly,
the mechanical effect of ultrasonic wave improves heterogeneous reactions of polysaccharides in terms of smoother experimental conditions. Secondly, due to the easy formation
of transient reactive species new transformation can be designed [30]. After the degradation of PLFP induced by ultrasonic treatment the microstructure was rearranged, which led
to the formation of smaller particle aggregation and was in
agreement with the result of Zhao et al. [31].

Fig. 4. Scanning electron micrographs of PLFP IeIV. A: PLFP I, B: PLFP II, C: PLFP III and D: PLFP IV.

272

B. Yang et al. / Polymer Degradation and Stability 93 (2008) 268e272

Acknowledgements
The financial support provided by National Natural Science
Foundation of China (No. 30425040), 11th five-year National
Key Technology R&D Program (Nos. 2006BAD27B03 and
2006BAD27B04) and Guangzhou Scientific Research Foundation (Nos. 2004Z1-E0061 and 2005Z2-E0151) is highly appreciated. We are grateful to Dr. Andrew Macnish of University
of California for his assistance in the improvement of the revised manuscript.
References

Fig. 5. The UVevis spectra of PLFP IeIV at a range of 195e550 nm. 1: PLFP
I, 2: PLFP II, 3: PLFP III and 4: PLFP IV.

3.4. UVevis spectra


Absorption of UVevis radiation by a molecule is related to
excitation of the outer electrons [32]. UVevis absorption is an
effective tool for chemical characterization because an adequate analysis of UVevis spectra may provide very interesting
information on the chemical structure of an analyte. The UVe
vis spectra of PLFP IeIV are presented in Fig. 5. No apparent
differences existed among these four spectra at a range from
195 to 550 nm, which indicated that ultrasonic treatment
may not cause significant chemical modification of groups in
the PLFP chain. The UVevis spectra of these four PLFP samples showed a predominant peak at 199e200 nm while
a slowly decreasing profile at 260e280 nm might indicate
the occurrence of small amounts of proteins [33].

4. Conclusions
Ultrasonic power, time and temperature are the important
factors influencing the extraction of PLFP. The highest recovery rate (5.49 mg GE/g DW) of PLFP was achieved at 120 W
and 70  C for 20 min. Four PLFP samples were prepared by
hot-water and ultrasonic extractions. The DSC analysis
showed that the To, Tp, Tc and DH of PLFP I were lower
than those of PLFP IIeIV, which meant that ultrasonic treatment caused changes in the microstructure of PLFP. PLFP II
had the highest DH (8.0 J/g) and exhibited two endothermic
peaks which were unlike the other three samples. Scanning
electron micrographs indicated the presence of many aggregated particles in PLFP III and IV. However, no apparent differences existed among the UVevis spectra of these four
PLFP samples at a range of 195e550 nm. Further investigations should be considered to obtain more understanding of
the effect of ultrasonic treatment on the chemical structure
of PLFP.

[1] Schepetkin IA, Quinn MT. Int Immunopharmacol 2006;6:317.


[2] Wang Q, Fang Y. J Chromatogr B 2002;812:219.
[3] Deters A, Dauer A, Schnetz E, Fartasch M, Hensel A. Phytochemistry
2001;58:949.
[4] Hromadkova Z, Ebringerova A. Ultrason Sonochem 2003;10:127.
[5] Vinatoru M, Toma M, Radu O, Filip PI, Lazurca D, Mason TJ. Ultrason
Sonochem 1997;4:135.
[6] Tsochatzidis NA, Guiraud P, Wilhelm AM, Delmas H. Chem Eng Sci
2001;56:1831.
[7] Velickovic DT, Milenovic DM, Ristic MS, Veljkovic VB. Ultrason Sonochem 2006;13:150.
[8] Zhou C, Ma H. J Agric Food Chem 2006;54:2223.
[9] Mislovicova D, Masarova J, Bendzalova K, Soltes L, Machova E. Ultrason Sonochem 2000;7:63.
[10] Zhang M, Zhang L, Cheung PCK, Ooi VEC. Carbohydr Polym
2004;56:123.
[11] Jiang YM, Zhang ZQ, Joyce DC, Ketsa S. Postharvest Biol Technol
2002;26:241.
[12] Yang B, Wang JS, Zhao MM, Liu Y, Wang W, Jiang YM. Carbohydr Res
2006;341:634.
[13] Navarini L, Gilli R, Gombac V, Abatangelo A, Bosco M, Toffanin R.
Carbohydr Polym 1999;40:71.
[14] Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. Anal Chem
1956;28:350.
[15] Zhao MM, Yang B, Wang JS, Li BZ, Jiang YM. Food Chem
2006;98:539.
[16] Vinatoru M. Ultrason Sonochem 2001;8:303.
[17] Li J, Guo S, Li X. Polym Degrad Stab 2005;89:6.
[18] Sivakumar M, Pandit AB. Ultrason Sonochem 2001;8:233.
[19] Hemwimol S, Pavasant P, Shotipruk A. Ultrason Sonochem 2006;13:543.
[20] Palma M, Barroso CG. Anal Chim Acta 2002;458:119.
[21] Lazaridou A, Biliaderis CG, Izydorczyk MS. Food Hydrocolloids
2003;17:693.
[22] Lazaridou A, Biliaderis CG. Food Hydrocolloids 2004;18:933.
[23] Chung HJ, Lee EJ, Lim ST. Carbohydr Polym 2002;48:287.
[24] Prasertsan P, Dermlim W, Doelle H, Kennedy JF. Carbohydr Polym
2006;66:289.
[25] Lionetto F, Maffezzoli A, Ottenhof MA, Farhat IA, Mitchell JR. J Food
Eng 2006;75:258.
[26] Wasikiewicz JM, Yoshii F, Nagasawa N, Wach RA, Mitomo H. Radiat
Phys Chem 2005;73:287.
[27] Spengler J, Jekel M. Ultrasonics 2000;38:624.
[28] Szu SC, Zon G, Schneerson R, Robbins JB. Carbohydr Res 1986;152:7.
[29] Wu HC, Shen FW, Hong X, Chang WV, Winet H. Biomaterials
2003;24:871.
[30] Kardos N, Luche JL. Carbohydr Res 2001;332:115.
[31] Zhao LJ, Li J, Guo SY, Du Q. Polymer 2006;47:2460.
[32] Pons MN, Le Bonte S, Potier O. J Biotechnol 2004;113:211.
[33] Lange R, Balny C. BBA-Protein Stuct Mol Enzym 2002;1595:80.