Chemical Papers

https://doi.org/10.1007/s11696-020-01140-2

ORIGINAL PAPER

Comparison of different extraction techniques and optimization

of the microwave‑assisted extraction of saponins from Aralia elata
(Miq.) Seem fruits and rachises
Ya‑qin Tian1 · Heng‑tian Zhao2 · Xiu‑ling Zhang1 · Wen‑tao Zhang1 · Xiao‑chen Liu1 · Shi‑han Gao1

Received: 28 October 2019 / Accepted: 17 March 2020

© Institute of Chemistry, Slovak Academy of Sciences 2020

Abstract
The total saponins from Aralia elata (Miq.) Seem fruits and rachises were isolated by microwave-assisted extraction (MAE),
ultrasonic extraction, heat reflux extraction, and soxhlet extraction. These techniques were compared from four perspec-
tives: extraction kinetics, saponin yield, disintegration to the raw materials, and antioxidant activity. MAE had the highest
extraction efficiency, shortest time, and greatest disintegration to the raw materials. The extracts obtained with MAE had the
highest antioxidant activities, as the DPPH free radical scavenging abilities of the fruit and rachis extracts were 70.86 ± 1.33
and 50.85 ± 1.68 mg VC g−1 md, respectively. The MAE parameters were optimized by the response surface methodology.
The optimum conditions (92% and 91% ethanol, solid-to-liquid ratios of 1:30 and 1:22, microwave powers of 530 W and
400 W, microwave times of 40 s and 54 s) provided extraction yields of 2.98 ± 0.037 mg g−1 and 0.94 ± 0.026 mg g−1 from
the fruits and rachises, respectively.

Keywords Saponins · Microwave-assisted extraction · Antioxidant · Optimization · Aralia elata (Miq.) Seem

Introduction

Aralia elata (Miq.) Seem, commonly known as “ci long

ya” and “ci lao ya”, is a small edible and medicinal per-
ennial deciduous tree belonging to Araliaceae Aralia (Gao
Electronic supplementary material The online version of this
et al. 2019; Zhang et al. 2013c). The fruiting period is from
article (https​://doi.org/10.1007/s1169​6-020-01140​-2) contains
supplementary material, which is available to authorized users. August to September, and the fruits are black, 4 mm in diam-
eter, and generally spherical, although they have five edges
* Xiu‑ling Zhang (Shikov et al. 2016). Aralia elata is widely distributed in
zhangxiuling1968@126.com
northeastern China, North Korea, South Korea, Japan, and
Ya‑qin Tian Russia (Zhang et al. 2006, 2013b). Aralia elata is rich in a
tianyaqin19920902@126.com
variety of chemical components. The chemical constituents
Heng‑tian Zhao isolated from the root bark, stem bark, and leaves of A. elata
zhaohengtian@163.com
include saponins, flavonoids, fats, sugars, aromatic oils, vol-
Wen‑tao Zhang atile oils, amino acids, proteins, and inorganic compounds.
zhangwentaoneau@126.com
Many studies have shown that these constituents have vari-
Xiao‑chen Liu ous biological activities, such as antitumor (Liu et al. 2016;
282026796@qq.com
Otsuka et al. 2014), antioxidative (Wang et al. 2018; Zhang
Shi‑han Gao et al. 2013a), anti-inflammatory (Suh et al. 2007), and liver-
1667057510@qq.com
protective (Luo et al. 2015). However, there are few reports
1
College of Food Science, Northeast Agricultural University, on the fruits and rachises, and no methods of processing or
Harbin 150030, Heilongjiang, China utilizing the fruits and rachises have been reported.
2
Northeast Institute of Geography and Agroecology, Chinese Traditional extraction and isolation methods, such as
Academy of Sciences, Harbin 150030, Heilongjiang, China immersion, percolation, reflux, Soxhlet, and supercritical

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fluid extraction, face certain challenges in the extraction of
saponins due to their high energy consumption, long extrac-
tion times, and high equipment costs (Jesus et al. 2019; Wei
and Yang 2015). Microwave-assisted extraction (MAE) is a
green extraction technology that is widely used in labora-
tories. The microwave radiant energy is transferred to the
interior of the raw material through the extraction medium.
Since the raw material absorbs microwave energy, its inter-
nal temperature rises rapidly. Increased internal pressure
causes cell rupture, resulting in the rapid dissolution of the
target components (Rodriguez-Jasso et al. 2011). Many
studies have shown that the MAEs of saponins (Ramli et al.
2019), phenolics (Kwiatkowski et al. 2020), flavonoids
(Zhao et al. 2020), polysaccharides (Chen and Xue 2019;
Mirzadeh et al. 2020), and volatile oils (Chouaibi et al. 2019)
were superior to traditional extraction methods. At present,
MAE is widely used in the production of spices, seasonings,
natural products, cosmetics, Chinese herbal medicines, and
natural pigments.
The objective of this study was to develop and utilize the
fruits and rachises of A. elata, increasing the economic value
of this plant. In the first part of this study, the efficiencies
of MAE, ultrasonic extraction (UE), heat reflux extraction
(HRE), and soxhlet extraction (SE) were compared based on
their extraction kinetics and saponin yields. In the second
part, the degree of damage to the raw materials during the
extraction was determined by scanning electron microscopy.
The antioxidant activities of the extracts were also deter-
mined. Finally, the effects of MAE on the extraction yields
of saponins from A. elata fruits and rachises were evaluated.
The process parameters, including ethanol concentration,
solid–liquid ratio, extraction time, and microwave power,
were optimized.
Experimental
Chemicals and reagents
Methanol ­(CH 3OH), absolute ethanol (EtOH ≥ 99.8%),
vanillin, potassium persulfate (­K 2 S 2 O 8 ), acetic acid
­(CH3COOH), perchloric acid ­(HClO4), potassium ferricya-
nide ­(K3[Fe(CN)6]), and F
­ eSO4 were purchased from the
Shanghai Yuanye Biotechnology Corporation. Oleanolic
acid standard, 1,1-diphenyl-2-picrylhydrazyl (DPPH),
2,2’-azinobis(3ethylbenzothiaoline-6-sulphonic acid) diam-
monium salt (ABTS), and ascorbic acid (VC) were purchased
from Solarbio (USA).
Extraction procedures
The fruits and rachises of A. elata obtained from 10-year-old
trees were hand-harvested at optimum ripeness in August
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Chemical Papers
2018. Fruits and rachises were picked from the Chinese
Academy of Sciences in Harbin, China, and authenticated
by Prof. Xiu-ling Zhang from the Food College of Northeast
Agricultural University. Voucher specimens were depos-
ited in the Fruit and Vegetable Storage and Preservation
Laboratory, College of Food Science, Northeast Agricul-
tural University. The dried A. elata fruits nd rachises were
crushed with a disintegrator (JP1000B, Zhejiang, China) and
sieved (60 mesh). The obtained powder (5.0 g samples) was
extracted by different extraction methods (MAE, UE, HRE,
and SE).
Microwave‑assisted extraction (MAE)
MAE was carried out in a MARS-II (1000 W, 2450 MHz)
microwave-accelerated reaction system. The powder
(5.0 g) was accurately weighed into three-necked flasks.
For the comparison of different extraction methods, an
ethanol–water solution (80%) was added according to the
experimental design, and the samples were maintained at
room temperature for 30 min. A sample–solvent ratio of
1:20, microwave irradiation power of 350 W, irradiation
time of 90 s, and three extraction cycles were applied to the
samples. For optimization of the MAE of total saponins, the
extraction process was conducted under different conditions
including the following: ethanol concentration (60–100%),
solid–liquid ratio (1:5–1:80), irradiation time (10–60 s),
irradiation power (350–700 W), and number of extraction
cycles (1–5).
Ultrasonic extraction (UE)
The powder (5.0 g) was weighed into a triangular flask,
100 mL of 80% ethanol solution (80:20, v/v) was added, and
the mixture was allowed to stand for 30 min. The flask was
transferred to an ultrasonic bath (JB-031S, Shenzhen, China)
at 70 °C for 155 min. This process was repeated twice for a
total of three cycles.
Heat reflux extraction (HRE)
A sample of the powder (5.0 g) and 100 mL of 80% etha-
nol–water solution (80:20, v/v) were mixed in a round-bot-
tomed flask. The flask was placed in a water bath, and the
mixture was refluxed at 90 °C for 2 h. This process was
repeated twice for a total of three cycles.
Soxhlet extraction (SE)
A sample of the powder (5.0 g) was weighed into a Soxhlet
apparatus and extracted with 100 mL of ethanol–water solu-
tion (80:20, v/v) at 90 °C for 6 h. This process was repeated
twice for a total of three cycles.
Chemical Papers
Determination of the extraction yield
The yields of saponins from A. elata fruits and rachis were
determined according to the method described by Hiai
et al. (1976). Twenty microlitres of the extract was added
to a test tube, and the ethanol was evaporated in a water
bath at 70 °C. Then, 5% vanillin–glacial acetic acid solu-
tion (0.2 mL) and perchloric acid (0.8 mL) were added.
The mixtures were heated in a constant temperature water
bath at 60 °C for 15 min. The samples were transferred to
an ice bath, after 5 min, 5 mL of glacial acetic acid was
added, and the absorbance was measured at 560 nm using
an ultraviolet–visible spectrophotometer (Cary 50 Bio,
Victoria, Australia). The saponin yield was calculated as
mg of oleanolic acid equivalents per 1 g of dried sample.
Scanning electron microscopy (SEM)
The morphologies of the dry residues obtained from dif-
ferent extraction methods were assessed using a Maia-3-
Xmh field scanning electron microscope (Tescan3, Shang-
hai, China). The experimental method was based on that
described by Yan et al. (2010) and Lou et al. (2010) with
some modifications. The particles were fixed on tape and
sputter-coated with gold, and then, they were assessed by
SEM using an accelerating voltage of 15.0 kV (20 μm,
2000× magnification).
Antioxidant activity
The DPPH assay was conducted following the method
of Fattouch et al. (2007) with slight modifications. Two
millilitres of the sample extract and 2 mL of DPPH solu-
tion were reacted at room temperature for 30 min in the
dark, and the absorbance at a wavelength of 517 nm was
measured, with 70% methanol used as a blank control.
The results are expressed in mg ­VC equivalents per 1 g of
dried sample.
The FRAP assay was conducted as described by Ben-
zie and Strain (1996) and Huang et al. (2005) with slight
modifications. The absorbance at 593 nm was measured,
and 70% methanol solution was used as a blank control.
The results are expressed in mg of ­VC equivalents per 1 g
of dried sample.
The ABTS assay was conducted as described by Re
et al. (1999); namely, 3 mL of ­ABTS∙+ working solution
and 30 μL of sample extract were mixed at 30 °C for 6 min
in the dark, and the absorbance at 743 nm was measured;
methanol was used as a blank control. The results are
expressed in mg of Trolox equivalents per 1 g of dried
sample.
Box–Behnken design for extraction optimization
Optimization experiments for the MAE of total saponins
from A. elata (Miq.) Seem fruits and rachises were per-
formed by the response surface methodology (RSM) using
a three-level (− 1, 0, 1) four-factor Box–Behnken design.
Based on single-factor experiments, the ethanol concentra-
tion (X1), ratio of solid to liquid (X2), microwave irradiation
power (X3), and microwave irradiation time (X4) had a sig-
nificant effect on the yield of saponins. The four factors were
selected as independent variables, while the saponin yield
(Y) was the dependent variable (Tables 3, 4). The results
were analysed using the software Design Expert 8.0.6 Trial
(Stat-Ease, Inc.). A second-order polynomial model was fit-
ted to evaluate the experimental results:
k k k−1 k
∑ ∑ ∑ ∑
Y = 𝛽0 + 𝛽i X i + 𝛽ii X2ii + 𝛽ij Xi Xj
i=1 i=1 i j
where Y is the response variable; Xi and Xj are independent
variables; and β0, βi, βii, and βij are regression coefficients.
Statistical analysis
All experiments were repeated three times, and the data are
expressed as the mean ± standard deviation. The data were
subjected to Duncan’s test using one-way analysis of vari-
ance (ANOVA) in the statistical analysis software SPSS 20.0
to analyse the correlations between data.
Results and discussion
Extraction kinetics
The kinetics of the extraction of saponins from the fruits
and rachises of A. elata using different extraction methods
are shown in Fig. 1. In MAE, the yield of total saponins
increased sharply as the extraction time was increased
up to 50 s. When the irradiation times were 50–60 s, the
yields of total saponins decreased. The saponins dis-
solved in the solvent underwent chemical changes due to
the prolonged irradiation time and increased temperature
in the system, and the dissolution rate could not make up
for the rate of chemical changes, resulting in a decrease
in the extraction efficiency of the saponins (Chen et al.
2008). In UE, the yield of saponins increased sharply as
the extraction time was increased to 55 min. When the
extraction time exceeded 55 min, the yield of saponins
began to greatly decrease with increasing time because
over time, the energy carried by the high frequency ultra-
sonic waves will seriously damage the dissolved sapo-
nins, causing most of the saponins to undergo chemical
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Fig. 1  Extraction kinetics of total saponins from Aralia elata (Miq.) Seem fruits and rachises by different extraction techniques. The values are
the means of three replicates for each extraction. Bars on the symbols indicate the standard deviation of the means

Table 1  Comparison of different extraction techniques of saponins from Aralia elata (Miq.) Seem fruits and rachises
Material Extraction Temperature Power (W) Extraction time (s) Extraction yields (mg/g) Degree of damage
(°C)

Fruit MAE 40 350 50 1.22 ± 0.039a Serious damage

UE 70 200 3.3 × 103 0.75 ± 0.024c Obvious damage
SE 90 1200 1.44 × 104 1.11 ± 0.016b Slight damage
HRE 90 1000 3.6 × 103 1.12 ± 0.026b No damage
Rachis MAE 40 350 50 0.78 ± 0.030a Serious damage
UE 70 200 3.3 × 103 0.54 ± 0.010b Obvious damage
SE 90 1200 1.44 × 104 0.75 ± 0.018a Slight damage
HRE 90 1000 3.6 × 103 0.74 ± 0.013a No damage

Mean ± standard deviation (n = 3). Different lowercase letters indicate significant differences (p < 0.05)

changes (Xie et al. 2019; Zeković et al. 2017). In HRE The yields of total saponins from the MAE of A. elata
and SE, the saponin yield initially increased and finally fruits and rachises were 1.22 ± 0.039 and 0.78 ± 0.030 mg g−1,
remained constant. respectively, which were significantly higher than those of SE

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Fig. 2  Scanning electron Rachis Fruits

microscopy images (20 μm,
×2000 magnification) of the raw RW RW
material (RW) and the Aralia
elata (Miq.) Seem processed
by four different extraction
methods, heat reflux extraction
(HRE), soxhlet extraction (SE),
ultrasonic extraction (UE), and
microwave-assisted extrac-
tion (MAE). The left panels
are images of the Aralia elata
(Miq.) Seem fruit residues, and HRE HRE
the right panels are images of
the Aralia elata (Miq.) Seem
rachis residues

SE SE

UE UE

MAE MAE

(1.11 ± 0.016 and 0.75 ± 0.018 mg g−1), HRE (1.12 ± 0.026
and 0.74 ± 0.013 mg g −1), and UE (0.75 ± 0.024 and
0.54 ± 0.010 mg g−1). The results were consistent with those
of Dong et al. (2014). In addition, MAE had no temperature
requirement and could be carried out at room temperature,
which saved more energy than SE, HRE, and UE. Compared
with SE and HRE, MAE required a lower power and con-
sumed less power (Table 1). The extraction time of MAE was
significantly less than those of SE, HRE, and UE. Therefore,
MAE had the highest extraction efficiency.
SEM observations
The raw material (RW) and extraction residues from MAE,
UE, HRE, and SE were observed by SEM to evaluate the
degree of disintegration to the A. elata fruit and rachis pow-
ders by the four different extraction methods and to explore
the extraction mechanisms (Table 1 and Fig. 2). MAE caused
a large number of pores on the surface of the fruit residue and
serious cracks on the surface of the rachis residue. The sapo-
nins in the samples were rapidly transferred to the solvent

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Table 2  Analysis of the Material Extraction
antioxidant capacities of the
extracts of Aralia elata (Miq.)
Seem fruits and rachises
Fruit MAE
UE
SE
HRE
Rachis MAE
UE
SE
HRE
Mean ± standard deviation (n = 3). Different uppercase letters indicate significant differences between val-
ues in the same column (p < 0.05); different lowercase letters indicate significant differences between val-
ues in the same row (p < 0.05)
through the channels and cracks, increasing the saponin
extraction efficiency (Ren et al. 2015). Following UE, the
surfaces of the fruit and rachis residues showed obvious dis-
integration, and the mechanical damage caused by the cavita-
tion generated by the ultrasonic waves allowed the transfer
of the saponins to the solvent (Da Porto and Natolino 2018).
However, the degree of disintegration to the sample was less
than that caused by microwaves, and the extraction efficiency
was lower than that of MAE. In SE, the saponins were trans-
ferred to the solvent by osmosis and dissolution (Yan et al.
2010), and the surfaces of the residues were slightly cracked.
In HRE, the surfaces of the samples were flat and had few
voids, and the surfaces were not significantly different from
the surface of the RW. Therefore, MAE caused the greatest
disintegration to raw materials, resulting in a high extraction
efficiency and a short extraction time.
Determination of the antioxidant activities
The effects of the four extraction methods on the antioxidant
activities of the total saponins were evaluated by measuring
the ferric-reducing antioxidant power, DPPH free radical
scavenging activity, and ABTS free radical scavenging activ-
ity. ­DPPH· is a stable free radical that is often used in free
radical scavenging assays (Kedare and Singh 2011). For the
fruit extracts, the DPPH free radical scavenging activities of
the total saponins obtained by the four extraction methods
were in the following order: MAE > HRE > SE > UE. By
this measure, MAE was superior to the other three extrac-
tion methods, most likely because the saponins obtained by
MAE released more hydrogen ions, which could bind to the
lone pair of electrons on the substrate, converting the DPPH
radicals into stable compounds (Sparg et al. 2004). The fer-
ric-reducing antioxidant power of the extract obtained by
MAE was 27.92 ± 1.02 mg VC g−1, which was significantly
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Chemical Papers
DPPH FRAP ABTS The units
of ABTS are mg
mg VC/g md mg Trolox/g
md The units of FRAP are mg VC/g md Trolox/g md
50.85 ± 1.68Aa 27.61 ± 1.05Ac 14.03 ± 1.76Ad
41.81 ± 1.57ABa 10.16 ± 1.58Cb 9.24 ± 1.48Bb
37.74 ± 1.08Ba 27.92 ± 1.02Ab 5.18 ± 1.57Bc
41.81 ± 1.27ABa 19.31 ± 1.17Bc 13.65 ± 0.21Ad
76.44 ± 0.75Aa 29.75 ± 1.37Ab 28.58 ± 0.87Bb
47.24 ± 1.38Da 10.86 ± 1.48Dc 7.86 ± 0.37Cd
68.11 ± 1.32Ca 17.78 ± 0.79Cc 32.33 ± 2.55Ab
70.86 ± 1.33Ba 20.47 ± 0.93Bb 5.32 ± 0.96Dc
higher than those of the extracts obtained by the other three
methods. The ABTS free radical scavenging activities of
the samples obtained by MAE were the highest, followed
by those obtained by SE, and UE and HRE resulted in lower
activities. The effects of the four extraction methods on the
antioxidant activities of the total saponins from A. elata
rachises were similar to those from the fruits (Table 2). In
conclusion, the antioxidant activities of the extracts were
better preserved by MAE, and MAE was the best method
for extracting the saponins from A. elata fruits and rachises.
Results of the selection of variables
The extractions of total saponins from A. elata fruits and
rachises were studied by choosing the following five key
parameters: ethanol concentration, solid–liquid ratio, irra-
diation time, irradiation power, and number of extraction
cycles. As can be seen from Fig. 3, the ethanol concentra-
tion, solid–liquid ratio, irradiation time, and irradiation
power had significant effects on the yields of total saponins
from A. elata fruit and rachis, but the number of extrac-
tion cycles did not. Figure 3a shows that the yields of
total saponins increased dramatically as the ethanol con-
centration was increased from 60 to 100%. As shown in
Fig. 3b, the yields of total saponins increased dramatically
with increasing solid-to-liquid ratios and reached maxi-
mum values at a solid-to-liquid ratio of 1:20. These results
show that a higher ratio of solid to liquid did not always
increase the yield of the target compound, which may
be due to the larger amount of solvent causing excessive
swelling of the raw material and increasing the absorp-
tion of the target compound (Sarvin et al. 2018). As the
microwave irradiation power was increased from 350 W
to 560 W, the yield of total saponins from A. elata fruits
significantly increased. The yield of total saponins from
A. elata rachises initially increased and then decreased
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Fig. 3  a Effects of ethanol

concentration on the yields
of total saponins. b Effects of
solid–liquid ratio on the yields
of total saponins. c Effects of
microwave irradiation power on
the yields of total saponins. d
Effects of microwave irradia-
tion time on the yields of total
saponins. e Effects of extraction
cycles on the yields of total sap-
onins. Different letters within
each column indicate significant
differences at p < 0.05

with increasing microwave irradiation power (Fig. 3c).
Figure 3d shows that the yields of total saponins initially
increased and then decreased. Microwave irradiation for
50 s increased the maximum yields to 1.58 ± 0.018 mg g−1
(fruit) and 0.63 ± 0.010 mg g −1 (rachis), and extending
this time led to lower yields. Re-extraction using fresh
solvent facilitates the complete dissolution of the target
compound (Sarvin et al. 2018). Interestingly, the effects of
the number of extraction cycles on the yields of total sapo-
nins from A. elata fruits and rachises were not significant.
Therefore, the ethanol concentration, solid–liquid ratio,
irradiation time, and irradiation power were selected as
the extraction parameters of the RSM design.
Optimization of the extraction of total saponins
The three-level four-factor Box–Behnken plan for the ethanol
concentration, solid–liquid ratio, irradiation time, and irradia-
tion power was designed using Design Expert 8.0.6 Trial (Stat-
Ease, Inc.). The plan and results are listed in Tables 3 and 4.
Table 5 summarizes the adjusted coefficients of determination
Adj-R2, which were 0.991 and 0.9643, respectively, indicating

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Table 3  Optimization Run Extraction conditions Experimental result

experience plan of RSM Box–
Behnken design for Aralia elata Concentration Radio of solid/ Microwave irra- Microwave irra- Extraction yield (mg/g)
(Miq.) Seem fruits of ethanol (%) liquid (mg/ml) diation power (W) diation time (s)

1 95 20 630 40 1.36 ± 0.036

2 95 20 560 50 2.92 ± 0.014
3 95 30 560 60 2.17 ± 0.020
4 95 20 630 60 2.49 ± 0.025
5 95 10 560 40 1.72 ± 0.027
6 95 10 560 60 2.56 ± 0.033
7 95 30 560 40 2.82 ± 0.018
8 95 20 560 50 2.93 ± 0.035
9 95 10 630 50 1.43 ± 0.039
10 95 20 560 50 2.83 ± 0.036
11 100 20 630 50 1.84 ± 0.035
12 100 20 560 60 2.47 ± 0.052
13 100 10 560 50 2.32 ± 0.038
14 95 30 490 50 2.17 ± 0.029
15 95 20 560 50 2.89 ± 0.027
16 95 20 490 60 1.29 ± 0.034
17 90 20 490 50 1.43 ± 0.028
18 95 20 560 50 2.76 ± 0.053
19 100 20 560 40 1.91 ± 0.033
20 90 30 560 50 2.04 ± 0.034
21 90 10 560 50 1.12 ± 0.013
22 90 20 560 40 2.26 ± 0.074
23 95 30 630 50 2.30 ± 0.050
24 90 20 560 60 1.18 ± 0.049
25 95 10 490 50 1.73 ± 0.035
26 100 20 490 50 1.72 ± 0.035
27 100 30 560 50 1.73 ± 0.025
28 95 20 490 40 2.56 ± 0.030
29 90 20 630 50 1.33 ± 0.028

Mean ± standard deviation (n = 3)

the high reliability of the model. The nonsignificant value of
the lack of fit (p > 0.05) demonstrated that the unknown factors
had little effect on the results. The fitted second-order polyno-
mial equation of the yields of total saponins from A. elata fruit
(Y1) and rachis (Y2) was as follows:
Y1 = 2867.40 + 218.43X1 + 195.57X2
− 378.96X1 X2 + 409.36X1 X4 − 372.82X2 X4
+ 598.35X3 X4 − 679.48X12 − 335.12X22
− 641.26X32 − 250.49X42
Y2 = 958.23 − 51.28X1 + 22.51X2 + 18.96X3 − 47.21X4
+ 139.23X1 X3 − 104.43X1 X4 − 138.70X2 X3
− 118.10X3 X4 − 39.22X12 − 142.10X22
− 156.30X32 − 116.95X42
For A. elata fruits, the linear effect of ethanol con-
centration (X1) showed a significant (p < 0.001) positive
effect on the total saponins, while the ethanol concentra-
tion quadratic term (X12) had a significant (p < 0.001) neg-
ative effect on the total saponins, which was followed by
the irradiation power quadratic term (X23) and solid–liquid
ratio quadratic term (X 22). As the ethanol concentration
was increased, the yield of total saponins from A. elata
fruits significantly increased. An increased total phenolic
content with an increased methanol concentration was
also reported (Belwal et al. 2016). For A. elata rachises,
the linear effect of the ethanol concentration (X 1) and
microwave irradiation time (X 4 ) had negative effects.
However, a positive interaction between the ethanol con-
centration (X 1) and irradiation power (X 3) was observed
(Table 5). Thus, it is better to use a shorter microwave
irradiation time to allow the utilization of a higher ethanol

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Table 4  Optimization Run Extraction conditions Experimental result

experience plan of RSM Box–
Behnken design for Aralia elata Concentration Radio of solid/ Microwave irra- Microwave irra- Extraction yield (mg/g)
(Miq.) Seem rachises of ethanol (%) liquid (mg/ml) diation power (W) diation time (s)

1 90 10 420 50 0.82 ± 0.016

2 95 30 490 50 0.59 ± 0.081
3 100 20 420 60 0.59 ± 0.048
4 90 20 490 50 0.68 ± 0.091
5 95 10 420 60 0.69 ± 0.017
6 95 20 420 50 0.94 ± 0.026
7 95 10 350 50 0.45 ± 0.027
8 95 20 420 50 0.94 ± 0.034
9 95 20 490 40 0.87 ± 0.027
10 95 20 490 60 0.52 ± 0.024
11 95 30 420 40 0.77 ± 0.014
12 100 20 420 40 0.88 ± 0.011
13 95 10 420 40 0.71 ± 0.087
14 90 20 420 60 0.93 ± 0.061
15 95 20 420 50 0.98 ± 0.012
16 95 30 420 60 0.63 ± 0.010
17 95 20 420 50 0.97 ± 0.082
18 95 20 350 60 0.73 ± 0.020
19 90 20 420 40 0.81 ± 0.021
20 100 20 490 50 0.90 ± 0.025
21 95 10 490 50 0.76 ± 0.024
22 95 20 350 40 0.61 ± 0.023
23 90 20 350 50 0.91 ± 0.027
24 95 30 350 50 0.84 ± 0.022
25 100 20 350 50 0.57 ± 0.067
26 100 30 420 50 0.73 ± 0.052
27 100 10 420 50 0.71 ± 0.014
28 95 20 420 50 0.96 ± 0.056
29 90 30 420 50 0.84 ± 0.046

Mean ± standard deviation (n = 3)

concentration. Similar results were reported by other
studies, such as Piechowiak et al. (2020).
According to regression model analysis and the actual
conditions, the optimum conditions for the determina-
tion of total saponins from A. elata fruits were as fol-
lows: 92% ethanol, solid-to-liquid ratio of 1:30, micro-
wave power of 530 W, and microwave time of 40 s; the
optimum conditions for the determination of the total
saponins from A. elata rachises were as follows: 91%
ethanol, solid-to-liquid ratio of 1:22, microwave power
of 400 W, and microwave time of 54 s. The optimum
conditions were further verified, and the experiment was
repeated three times. The yields of total saponins from
A. elata fruits and rachises were 2.98 ± 0.037 mg g −1
and 0.94 ± 0.026 mg g−1, respectively. The experimental
value was close to the predicted value (the similarity was
greater than 95%), and the validity of the model was also
demonstrated.
Conclusion
At present, A. elata fruits and rachises are considered
waste, and productive uses have not been developed.
Four extraction methods (MAE, UE, HRE, and SE) were
used to extract the total saponins from A. elata fruits and
rachises, and the extraction time of MAE was shorter,
while the extraction efficiency was higher. Scanning elec-
tron microscopy (SEM) images showed that the cracks
and pores produced by MAE on the surface of the raw
material facilitated the rapid dissolution of the saponins,
improving the extraction efficiency and extraction yield.
In addition, the extracts obtained by MAE showed good

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