Journal of Food Engineering 93 (2009) 59–65

Contents lists available at ScienceDirect

Journal of Food Engineering

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

Kinetic study of saponins B stability in navy beans under different

processing conditions
John Shi a,*, Sophia Jun Xue a, Ying Ma b, Dong Li c, Yukio Kakuda d, Yubin Lan e
a
Guelph Food Research Center, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Canada N1G 5C9
b
School of Food Science and Engineering, Harbin Institute of Technology, Harbin 150090, China
c
College of Engineering, China Agricultural University, Beijing, China
d
Department of Food Science, University of Guelph, Guelph, Canada N1G 2W1
e
USDA-ARS, College Station, TX 77845, USA

a r t i c l e i n f o a b s t r a c t

Article history: Saponins are rich in the legumes which are known to provide many health benefits for human beings.
Received 24 June 2008 Saponin B is the main component in the saponins group present in navy beans. The stability of saponin
Received in revised form 2 December 2008 B during food processing is a key issue in evaluating the quality and nutrition of food products. The effects
Accepted 29 December 2008
of different soaking and cooking methods and conditions on the stability of saponin B were investigated.
Available online 19 January 2009
The effects of the soaking process on saponin reduction followed a first order kinetic model. The soaking
time and the seed-to-water ratio significantly affected the stability of saponin B during the soaking pro-
Keywords:
cess. Short time soaking and lower seed-to-water ratio would keep more saponin B in the soaked beans.
Navy bean
Saponins
The cooking medium and methods greatly influenced saponin B degradation during cooking. Water–oil
Kinetic degradation mixed cooking media enhanced saponins stability in the seeds during the cooking process, as compared
Soaking to a water-only cooking medium. Combined soaking and ordinary cooking induced more saponin degra-
Thermal processing dation in ordinary cooked seed samples. An autoclave cooking method eliminated most of the saponin B
from the autoclaved beans.
Crown Copyright Ó 2009 Published by Elsevier Ltd. All rights reserved.

1. Introduction
There is a long-standing controversy about saponins’ functions
in foods because saponins have generally been considered as unde-
sirable antinutrient components in legumes (Gestetner et al., 1968;
Khalil and El-Adawy, 1994). However, recent extensive studies of
their biological activity in vivo have shown that saponins’ stronger
bioactivities are associated with several health benefits. The results
suggested that saponins in the diet have a wide spectrum of activ-
ity as antifungal and antibacterial agents, in lowering of blood cho-
lesterol, and inhibition of cancer cell growth (Rao and Sung, 1995;
Jeon and Sung, 2000; Berhow et al., 2000; Oh and Sung, 2001;
Matsuura, 2001; Lee et al., 2005; Wang et al., 2007). Legumes are
one of the richest and least expensive natural sources of saponin
in the human diet.
Saponins possess both water-soluble and lipid-soluble compo-
nents. They consist of a lipid-soluble nucleus, having either a ste-
roid or a triterpenoid aglycone structure, with one or more side
chains of water-soluble carbohydrates. Based on their aglycone
structure, saponins are generally categorized into three main
groups as groups A, B, and E (Heng et al., 2006). The main saponin
* Corresponding author. Tel.: +1 519 780 8035; fax: +1 519 829 2602.
E-mail address: shij@agr.gc.ca (J. Shi).
components in legumes are the group B saponins. The group B sap-
onins have the aglycone, sapogenol B, as sapogenin and which
make up a majority of the saponins in legumes (Gurfinkel and
Rao, 2002; Hubert et al., 2005; Shiraiwa et al., 1991). However,
legumes are generally subjected to a cooking process before they
are consumed. Soaking and cooking are common processing meth-
ods for bean products in the food industry. Processing methods and
operating conditions tend to modify the composition and availabil-
ity of nutrients in the finished products (Van der Poel, 1990; Della
et al., 1994). The stability of saponins in the beans undergoing pro-
cessing directly influences the quality and nutritional value of the
end products. With advancements in qualitative and quantitative
analysis techniques, the precise analyses of saponin contents in
processed legume products is becoming more and more significant
to the food industry. More systemic and detailed reports on this as-
pect are needed. The processing methods and operating conditions
could affect the stability of saponins in the final bean products. Our
knowledge about the extent and nature of saponins’ kinetic degra-
dation is incomplete. The objective of the present study therefore
was to determine the degradation kinetics for saponins in beans
under various processing procedures that are commonly used in
the food industry, i.e. different soaking conditions and cooking
methods, and the effects of various thermal process methods and
conditions on the characteristics of the saponins during different

0260-8774/$ - see front matter Crown Copyright Ó 2009 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2008.12.035
60 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65
process stages using navy beans as a model material. This research
focused on the major saponins B in navy bean seeds.
2. Materials and methods
2.1. Plant materials
Dry raw navy bean (Phaseolus vulgaris L.) seeds were obtained
from the H.J. Heinz Company Canada Ltd. The dry beans were first
screened to get a uniform size free from broken or spoiled seeds.
The selected seeds were stored in a refrigerator at 4 °C in airtight
containers to prevent moisture absorption before applying various
treatments.
2.2. Processing methods
The processing methods used to study the kinetic degradation
of saponins were soaking, ordinary cooking, autoclaving, and their
combinations. In the soaking process, navy beans were soaked in
distilled water at different seed-to-water ratios with different
soaking times. In the ordinary cooking processes, effects of differ-
ent cooking media on the changes in saponins in the cooked unsoa-
ked and soaked beans were investigated. Comparisons of different
cooking methods on the degradation of saponins during the ther-
mal processes were determined. The experiments were prepared
and tested in triplicate in each step. The initial saponin B content
in the unprocessed dried navy beans was used as a control to com-
pare with the treated navy bean samples.
2.2.1. Soaking
Navy bean seeds were soaked in distilled water (pH 6.2) for 3, 6,
9, or 12 h at room temperature, respectively. To avoid the possible
problem that the soaking water may cause germination in the
seeds, the dry seed to soaking water ratio must be selected such
that the beans are completely covered by water. Therefore, the ra-
tios of dry beans to soaking water (called seed-to-water ratio), 1:3
and 1:7 (w/w), were chosen. Any incompletely soaked seeds were
discarded. The soaked bean seeds were rinsed in distilled water,
and then air-dried in a fume hood for 24 h at room temperature.
The dried beans were immediately milled into flour (8-mesh size)
and stored in dark brown polyethylene bottles at 30 °C until fur-
ther extraction and analysis.
2.2.2. Ordinary cooking
Two cooking media, distilled water and water–oil mixture (1:1,
v/v), were selected to study the effects of different cooking media
on the kinetic degradation of the saponins B group under different
cooking times. The ratio of seed-to-cooking media was fixed at the
1:4 ratio (cooking media is four times the weight of soaked seeds)
that is normally used in the traditional bean cooking process. After
soaking 50 g dry navy beans in distilled water at the seed-to-water
ratio of 1:3 for 6 or 12 h at room temperature, the soaked bean
seeds were rinsed with distilled water and dried with a paper towel,
then put in round mouth tall beakers fitted with condensers. After
adding 200 g distilled water or water–oil mixture, the samples were
cooked on a hot plate for 15, 25, or 35 min at 100 °C. The cooking
times chosen for this study are commonly used in the food industry
for bean processing. Cooking of unsoaked control seeds was also
performed by the same treatment method as described above. After
cooking, the medium was discarded and the beans were air-dried
overnight at room temperature. The dried cooked bean seeds were
ground into flour (8-mesh) while being cooled with liquid nitrogen
to avoid thermally induced degradation generated during milling.
The bean flours were stored in dark brown polyethylene bottles
at 30 °C, pending further extraction and analyses.
2.2.3. Autoclaving (pressure) cooking
The unsoaked and soaked (soaking for 6 h at a seed-to-water ra-
tio of 1:3) seeds were autoclaved (MRH Biotechnical Ltd., Boston,
MA, USA) in distilled water at 1.41 kg/cm2 pressure at a tempera-
ture of 121 °C for 15, 25, or 35 min at the seed-to-cooking media
ratio of 1:4 (w/w). The cooking times and ratios of seed-to-cooking
media used in the autoclave were the same as those used for ordin-
ary cooking. After pressure cooking, the media were discarded, and
the seeds were air-dried at room temperature, and then subse-
quently milled into flour (8-mesh size) with liquid nitrogen to
avoid mechanically induced thermal degradation during milling.
2.3. Extraction of saponins B
The saponins B in different processed navy bean seeds and con-
trol unprocessed seeds were extracted according to the method of
Khalil and Ei-Adawy (1994) with some modifications. Five grams
samples of bean flour from each of the different processes were ex-
tracted twice by refluxing for 4 h with 50 mL of 70% (v/v) aqueous
ethanol in a water bath at 60 °C. The extracts were centrifuged at
3000 rpm for 15 min. The supernatants were collected and dried
in a rotary evaporator at 45 °C for further hydrolysis. The dried
supernatants were dissolved in 8 mL of 1.5 N hydrochloric acid–
methanol solutions and hydrolyzed for 2.5 h at 75 °C. The hydroly-
sis mixtures were cooled, and then purified and concentrated by
solid phase extraction (SPE). Seven millilitres of hydrolyzed sam-
ples were passed through an SPE extraction cartridge (Strata
C18-E; 55 lm, 70 Å; 500 mg/6 mL; Phenomenex, USA), precondi-
tioned with methanol (5 mL) and distilled water (5 mL), and subse-
quently rinsed with 5 mL of distilled water to remove unbound
material. The fraction containing sapogenol B was eluted with
5 mL methanol. The fraction was then filtered through a 0.45 lm
syringe filter for HPLC analysis.
2.4. HPLC analysis
The total content reductions in saponins B were evaluated by
quantifying their common hydrolysate, sapogenol B. The reverse
phase HPLC (Agilent 1100, Scientific Equipment Source, ON, Can-
ada) with a UV detector was used to analyze the quantitative vari-
ations of saponins B at different conditions during soaking,
ordinary cooking, and autoclave. A Nova-Pak C18 column
(3.9  150 mm, 4 lm particle size; waters, USA) was used for sep-
aration. The mobile phase consisted of 75% acetonitrile and 25%
deionized water containing 0.05% TFA. The system was run isocrat-
ically at a flow rate of 1 mL/min. The sample injection volume was
10 lL and UV absorbance was monitored at 210 nm. Chromato-
grams were recorded and integrated with Agilent Chemstation
software. In order to quantify the saponin B, standard sapogenol
B (ChromDex, CA, USA) was prepared at 1 mg/mL (1000 ppm) in
methanol as a stock solution. Serial dilution of the stock solution
using methanol was performed to prepare series standard working
solutions with concentrations from 10 to 500 lg/mL. The represen-
tative chromatogram of sapogenol B standard and sapogenol B
obtained from navy bean samples are illustrated in Fig. 1A and B.
All analyses were carried out in triplicate and reported on a dry
matter basis.
2.5. Kinetic modeling of degradation
In comparison with the effects of different process methods and
conditions on saponins B contents in processed beans, the kinetic
phenomenon of saponins B under different conditions was demon-
strated by the plot of the first order degradation. The degradation
rate constant (k) was calculated as the slope of the linear plot. A
 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65
Fig. 1. (A) Representative chromatogram of standard sapogenol B (1000 lg/mL). (B) Representative chromatogram of sapogenol B obtained from navy bean samples by
hydrolyzing group B saponins.
first-order kinetic model was applied for characterizing saponins B
stability according to the following reaction:
dC
 ¼ kC
dt
The kinetic degradation of saponins to treatment times can be
obtained by integrating Eq. (1) as:
lnðC=C 0 Þ ¼ kt
where C0 is the initial saponins B content in unprocessed dry raw
navy bean seeds and C is the remaining saponins B contents in trea-
ted bean seeds after process time ‘t’. The k is the kinetic degradation
constant of saponins B (mg/g beans per min or h) and can be deter-
mined as the slope from plotting ln (C/C0) vs. t.
The reduction in saponin B contents was determined as a per-
centage of the decrease over the initial saponins B content in
unprocessed dry raw navy beans.
Soaking effects on saponins B reduction (%) by leaching =
(C0  Cs)/C0  100%
Cooking effects on saponins B reduction (%) by degradation and
leaching = (C0  Cs  Cc)/C0  100%, where C0 is the initial saponins
B content (mg/g) in unprocessed raw navy bean, Cs is the saponins
B content in the soaked seed samples, and Cc is the saponins B con-
tent in the cooked seed samples.
2.6. Statistical analysis
All experiments were conducted using factorial experimental
designs and analyzed in triplicate with a completely randomized
design. All statistical analyses were done by using SAS 8.2 software
from SAS Institute Inc., Cary, NC, USA. Kinetic data were analyzed
by regression analysis. ANOVA was used to determine the differ-
ence and interaction effects on saponin B contents among different
treatments such as the main factors and their combined effects,
including the various soaking times, seed-to-water ratios, and dif-
Table 1
The effects of soaking conditions on saponins B contents in navy beans.
Soaking time (h) Seed-to-water: 1:3
Saponins B contents (mg/g)
0 7.62 ± 0.19a
3 7.58 ± 0.17a
6 7.55 ± 0.14a
9 7.26 ± 0.13b
12 7.14 ± 0.16c
Means with the same superscript letters are not significantly different at p < 0.05 levels.
‘‘–” is not applicable.
61
ferent cooking media and cooking methods. Duncan’s multiple-
range comparison tests were conducted to determine the signifi-
cant effects of different processes on saponins B content of the
ð1Þ unprocessed and variously processed seed samples. Statistical sig-
nificance was accepted at p < 0.05.
3. Results and discussion
ð2Þ
3.1. Effects of soaking processes on degradation of saponins
The initial saponins B content in raw navy beans used in this
study was 7.62 ± 0.19 mg/g as shown in Table 1. The reduction of
saponins B amount occurs through a leaching process when the
beans are soaked in water. The results of Duncan’s tests showed
that 6 h or less soaking times did not significantly (p < 0.05) change
the saponin contents in the soaked seeds when the seeds were
soaked in distilled water at the seed-to-water ratio of 1:3 (w/w)
compared to their initial saponins B contents. However, increasing
the seed-to-water ratio to 1:7 at a soaking time of 6 h caused a sig-
nificant reduction of 3.1% saponins B content in the soaked seeds.
Moreover, when the soaking time was prolonged to 9 or 12 h, there
resulted significant reductions in the amount of saponins B in the
seeds soaked in distilled water at both the seed-to-water ratio of
1:3 and 1:7. The results suggested that the saponins B remained
stable for short soaking times with a limited amount of soaking
media. The dry beans initially absorb water to soften their hard
coats. Long time soaking treatments allow the beans to continu-
ously absorb water to breakdown the oligosaccharides and further
soften the bean tissue matrix to complete hydration (Upadhyay
and Garcia, 1988; Shimelis and Rakshit, 2007).
Saponins are soluble in water. The hydrated bean seeds allow
more water to penetrate more deeply into their matrix to release
saponins from the bean tissue matrix by simple diffusion (leach-
ing). These phenomena could explain the reason that the short
Seed-to-water: 1:7
Reduction (%) Saponins B contents (mg/g) Reduction (%)
– 7.62 ± 0.19a –
0.5 7.56 ± 0.10a 0.8
0.9 7.38 ± 0.11b 3.1
3.9 6.98 ± 0.13c 6.6
6.3 6.85 ± 0.11c 10.1
62 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65
time and less water (lower seed-to-water ratio) conditions did not
complete the hydration of beans hence did not generate significant
reductions of saponins contents. The highest reduction of 10.1%
saponins B content was observed in the seeds soaked for 12 h at
a seed-to-water ratio of 1:7. Simultaneously, foams were observed
in the soaking water when the seeds were soaked for more than
6 h, which indicated that saponin leaching occurred in the soaking
media. Saponins have the ability to foam in aqueous solutions. The
saponins B are more soluble in water due to their sugar chain
structure composition. Therefore, water leaching resulted in a loss
of saponins B (Gurfinkel and Rao, 2003; Shi et al., 2004) in soaked
beans during the soaking process. Longer soaking times and ex-
cesses water also allow a greater amount of sugar to dissolve,
resulting in the increased permeability of cell membranes and seed
coat to maximize the amount of saponins leached from the tissue
into the soaking media. However, ANOVA analysis showed that
no significant differences in saponin B contents were found in
the seeds soaked for 9 and 12 h at a seed-to-water ratio of 1:7. Pro-
longing soaking times would not appear to influence saponin
leaching after the bean tissues have been totally hydrated. The re-
sults suggest that the synergistic effects of soaking time and seed-
to-water ratios have more pronounced effects on the loss of sapo-
nin B contents during short soaking treatments.
The phenomenon of saponin kinetic mass transfer (leaching)
under different soaking conditions followed the first order kinetic
model with a correlation coefficient (R2) greater than 0.93
(Fig. 2). The results indicated that the seed-to-water ratios greatly
affected the stability of saponins B in the soaked seeds. The rate of
saponins B loss (in mg/g of sample) for soaked seeds at the seed-to-
water ratios of 1:3 and 1:7 were 0.0117/h and 0.0073/h, respec-
tively. This was evidence to confirm the synergistic effects of
seed-to-water ratio and soaking times on the reduction in the sap-
onins B contents.
From the kinetic mass transfer explanation for saponins B
leaching, the large amount of soaking media increases the diffusion
rate of the saponins, thus more water-soluble saponins were re-
leased from the bean tissue matrix into soaking media. Similar
trends were observed by previous researchers working with kidney
beans (Shimelis and Rakshit, 2007), moth beans (Khokhar and
Chauhan, 1986), chickpeas (Jood et al., 1986), and black grams
(Kataria et al., 1988). Kataria et al. (1989) reported that prolonging
the soaking time from 12 to 18 h did not have any significant influ-
ence on the saponin contents of mung-bean seeds during soaking.
However, in comparison with their range of 11–65% saponins
reduction in legumes by soaking processes (Khokhar and Chauhan,
1986; Kataria et al., 1989; Curl et al., 1985; Duhan et al., 2001;
0.00
-0.04
Ln (C/C0)
-0.08
-0.12
3 6
Soaking time (hour)
Fig. 2. Effects of soaking conditions on the kinetic degradation of saponins B in various soaked seeds (} is beans soaked at the seed-to-water ratio of 1:3, h is beans soaked at
the seed-to-water ratio of 1:7).
Drumm et al., 1990), our experimental results showed a lower
reduction in saponin contents (0.5–10.1%). The variability in the
observed amount of saponins reduction might be attributed to dif-
ferences in the bean tissues, pressing treatments, and/or to the ini-
tial saponin component compositions of these raw bean seeds
tissues.
3.2. Effects of different cooking media on the stability of saponins B
Navy beans are commonly cooked in water or cooked with meat
for meals. Therefore, the distilled water and water–oil mixture
were selected to study the effects of different cooking media on
the kinetic degradation of saponins B for the unsoaked and soaked
beans (soaked for 6 or 12 h at the seed to water ratio of 1:3) under
ordinary cooking conditions. Statistical results shows that the
seeds cooked in the distilled water or the water–oil mixture signif-
icantly reduced (p < 0.05) saponins B contents in all the cooked
seeds excepted for the samples (7.59 ± 0.17 mg/g) that are the
unsoaked seeds cooked in the distilled water for 15 min, as com-
pared to the initial saponins B contents (7.62 ± 0.19 mg/g). A com-
bination of soaking and cooking processes caused a high loss of
saponins B contents in the processed beans. The thermal treat-
ments have greater effects on saponins degradation because sapo-
nins B are a monodesmoside saponin and have a sugar chain linked
to the C-3 position of its aglycones (sapogenol B) with a hydrogen
atom at C-21 position (Heng et al., 2006). The linkage bond be-
tween sugar chain and sapogenol B would be broken, and the agly-
cones may also be decomposed when high energy is applied, such
as heating at higher temperatures. Therefore, thermolabile sapo-
nins degraded during cooking (Shi et al., 2004; Heng et al., 2006).
The water-soluble and oil-soluble characteristics of saponins B is
another reason to cause more saponins to be released from the
seed tissue matrix into the cooking media, resulted in a high de-
gree of degradation of saponins B. Therefore, the prolonged time
and excess thermal energy induced more saponins degradation
in the seeds. The cooking media also carried more saponins out
of the seed’s matrix during the cooking processes, especially when
the soaked seed was subjected to heating because of the softness
of the bean tissue permits the heat to more easily penetrate inside
of the seed’s matrix to degrade saponins B.
Fig. 3 shows the reductions of saponins in cooked bean seeds
under different cooking media. For the unsoaked seeds subjected
to ordinary cooking, the water–oil mixture showed more pro-
nounced effects on the reduction of saponins B as compared to
the water media. The oil media could maintain a higher tempera-
ture in the cooking media, enhancing the cooking media’s perme-
y 1:3 = -0.007x + 0.0227
2
R = 0.931
y 1:7 = -0.0111x + 0.0294
2
R = 0.9897
9 12
 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65 63

80 15 min 25 min 35 min

Saponins reduction (%)

60

40

20

0
water oil water oil water oil
unsoaked soaked 6 hrs soaked 12 hrs

Fig. 3. Effects of cooking media on the reductions of saponins B in cooked bean seeds.

ability into the matrix, and to drive degradation of saponins B dur-
ing the thermal process. However, progressive decreases in sapo-
nins B contents were found in the soaked seeds when cooked in
distilled water. One possible explanation for these phenomena lies
in the saponins B chemical structure. Saponins possess both lipo-
philic (aglycones) and hydrophilic (sugar chain) components. How-
ever, the saponins are more soluble in water than in lipid when the
sugar-chains are attached to the aglycones. The lipophilic structure
of the aglycone is also more stable than the sugar chain under ther-
mal treatment and hydration (Oakenfull and Sidhu, 1990; Oda
et al., 2003). Therefore, more water-soluble saponins have been
leached into the media. Simultaneously, products of decomposed
saponins that were subsequently oxidized by oxygen in the water
media caused a high degradation of saponin. More oxygen is pres-
ent in water than in oil media. The results suggest that the oil med-
ia is more favourable toward stabilizing saponins B in the beans, as
compared to water media traditionally used in the cooking process.
The degradation of saponins B in different cooking media under
thermal processes also followed the first-order kinetic degradation
model with a correlation coefficient (R2) greater than 0.94 (Figs. 4
and 5). The values of kinetic degradation rate constants (k) were
determined as the slope of the linear regression line from the plot
of the degradation of saponin contents in the samples vs. cooking
times. The values of k increased with increasing soaking time
and the cooking times. The results agreed with the conclusion that
the length of soaking and cooking are important factors that
greatly influenced the saponins B stability in the beans during
the process. Saponins degraded rapidly in the seeds after soaking
and were then subsequently subjected to thermal processes. In
comparing the kinetic degradation rate of saponins B in those
seeds cooked in water or a water–oil mixture, the values of the ki-
netic degradation rate constants were higher in the seeds first
soaked when followed by cooking in water than by cooking in a
water–oil mixture. These phenomena confirmed that oil media en-
hanced saponins B stability in beans during the cooking processes.
Similar results were noted in the saponin contents in moth bean
(Khokhar and Chauhan, 1986), edible dry beans (Oda et al.,
2003), soybean flour (Tarade et al., 2006), and kidney beans (Shi-
melis and Rakshit, 2007) during bean processing.
3.3. Effects of cooking methods on stability of saponins B
Table 2 shows a comparison of the cooking methods’ effects on
the kinetic degradation of saponins B in navy bean seeds during
thermal processing. The kinetic degradation of saponin did not fol-
lowed the first order kinetic model (R2 < 0.83) when the beans

0.2

-0.2
Y unsoaked = -0.015x + 0.2378
2
Ln (Cn/Co)

-0.4 R = 0.9656

-0.6
Y soaking 6hrs = -0.025x + 0.2823
2
-0.8 R = 0.9742

-1
Y soaking 12hrs = -0.0328x - 0.0244
-1.2 2
R = 0.9684
-1.4
15 20 25 30 35
Cooking time (min)

Fig. 4. Kinetic degradation of saponins in different combined soaking and ordinary cooking in water cooking media (} unsoaked seeds, h soaking for 6 h, 4 soaking for 12 h).
64 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65

-0.1

Ln (Cn/Co)
) -0.2

-0.3 Yunsoaked = -0.0155x + 0.2109

R 2 = 0.9971
-0.4
Ysoaking 6hrs = -0.0152x + 0.1045
2
R = 0.9401
-0.5

Y soaking 12hrs = -0.0161x + 0.0127

-0.6
R2 = 0.9437
15 20 25 30 35
Cooking time (min)

Fig. 5. Kinetic degradation of saponins in different combined soaking and ordinary cooking in water–oil cooking media (} unsoaked seeds, h soaking for 6 h, 4 soaking for
12 h).

Table 2
Comparison of ordinary and autoclave cooking effects on reduction of saponins B contents in treated seeds.

Cooking methods Ordinary Autoclave

Treatment Cooking time (min) Saponin (mg/g) Reduction (%) Saponin (mg/g) Reduction (%)
Unsoaked 0 7.62 ± 0.19a – 7.62 ± 0.19a –
15 7.59 ± 0.14a 0.39 2.02 ± 0.11i 73.49
25 6.86 ± 0.09c 9.97 N/D 100
35 5.62 ± 0.16f 26.25 N/D 100
Presoaked 6 h 0 7.55 ± 0.14a – 7.55 ± 0.14a –
15 7.05 ± 0.09b 6.62 0.58 ± 0.07k 92.32
25 5.12 ± 0.17d 32.19 N/D 100
35 4.28 ± 0.13g 43.31 N/D 100
Presoaked 12 h 0 7.14 ± 0.16b – 7.14 ± 0.16b –
15 4.12 ± 0.13e 42.30 N/D 100
25 3.29 ± 0.19h 53.92 N/D 100
35 2.14 ± 0.11i 70.03 N/D 100

Means with the same superscript letters are not significantly different at p < 0.05 levels.
‘‘–” is not applicable.
N/D is not detectable.

were cooked using an autoclave. The results showed that the deg-
radation of saponins B increased with increasing cooking time. For
the soaked and cooked beans in the ordinary cooking process, en-
hanced permeability of the seed membrane as a result of the soak-
ing processes improved the cooking process, which also had a
limitation on increasing the leached saponins in the boiling water.
The highest reduction in saponin contents in ordinary cooked sam-
ples was 70.03% when the seeds were first soaked for 12 h and then
subjected to cooking for 35 min. However, there were still some
saponin B contents remaining after 35 min cooking time. The low-
est content in the ordinary cooked seeds was 2.14 ± 0.11 mg/g
when the beans were soaked for 12 h and then cooked for
35 min. For autoclave cooking, no detectable amounts of saponin
B were found when the seeds were autoclaved for 25 min or longer.
Simultaneously, mashed cooked seeds were obtained after auto-
claving for 35 min because longer times under high pressure and
temperature caused the destruction of the navy bean seed matrix.
The thermally induced reduction of saponin contents was 73.49%
for an unsoaked bean autoclaved for 15 min, and 92.32% reduction
was found in the beans soaked for 6 h and then autoclaved for
15 min. The results suggested that autoclaving could destroy most
of saponins B in the beans, especially for the long-time-soaked
beans, even if only autoclaved for 15 min. The soaking process en-
hanced the effectiveness of the reduction of saponins B on the sub-
sequent ordinary cooking method. However, this effectiveness was
not as pronounced for the autoclave process. The differences in
saponin B degradation between ordinary cooking and autoclaving
might be attributed to the high energy level in the combination
of high pressure and high temperature in the autoclave, causing
those components such as aglycones and sugar to decompose,
resulting in a serious degradation of saponins in the beans. Because
aglycones possess a heat stable structure, they did not totally
decompose during ordinary cooking. Therefore, there were still
some amounts of saponin B that remained in the ordinary cooked
samples. These results are consistent with the reported studies by
Duhan et al. (2001), Sharma and Sehgal (1992), and Drumm et al.
(1990), all of whom reported that pressure cooking had a greater
reducing effect than ordinary cooking on the saponin contents of
pigeon peas and faba beans.
4. Conclusion
Processing methods and conditions greatly change the saponin
contents in beans during food preparation and processing. Kinetic
 J. Shi et al. / Journal of Food Engineering 93 (2009) 59–65
analysis results showed that the soaking times and seed-to-water
ratios significantly influenced the quantity of saponins B leached
out from the beans’ matrix during the soaking processes. Short
soaking times with less soaking media probably favoured stabiliz-
ing the saponins B in the beans. Cooking media and methods sig-
nificantly changed the saponin contents in different cooked
beans. Oil media seem to be less harmful on the degradation of
saponins B during the cooking processes. The pre-soaking process
enhanced subsequent thermally induced degradation of the sapo-
nins and caused a high loss of the contents during the cooking pro-
cesses. Autoclaving destroyed most of the saponins B in processed
beans. The lowest amount of saponin contents were found in pro-
cessed beans when beans that had been soaked for 6 h were sub-
jected to autoclaving for less than 15 min. These data would be
helpful for the food industry in finding a suitable method for main-
taining the content of health-promoting components or reducing
antinutrient substances in processed legume products.
References
Berhow, M.A., Wagner, E.D., Vaughn, S.F., Plewa, M.J., 2000. Characterization and
antimutagenic activity of soybean saponins. Mutation Research/Fundamental
and Molecular Mechanisms of Mutagenesis 448, 11–22.
Curl, C.L., Price, K.R., Fenwick, G.R., 1985. The quantitative estimation of saponins in
pea (Pisum sativum L.) and soya (Glycine max). Food Chemistry 18, 241–250.
Della, V., Balle, G., Quillien, L., Gueguen, J., 1994. Relationships between processing
conditions and starch and protein modifications during extrusion-cooking of
pea flour. Journal of Science and Food Agriculture 64, 509–517.
Drumm, T.D., Gray, J.I., Hosfield, G.L., Uebersax, M.A., 1990. Lipid, saccharide,
protein, phenolic acid and saponin contents of flour market classes of edible dry
beans as influenced by soaking and canning. Journal of Science of Food 51, 285–
297.
Duhan, A., Khetarpaul, N., Bishnoi, S., 2001. Saponin content and trypsin inhibitor
activity in processed and cooked pigeon pea cultivars. International Journal of
Food Sciences and Nutrition 52, 53–59.
Gestetner, B., Birk, Y., Tencer, Y., 1968. Fate of ingested soybean saponins and the
physiological aspect of their haemolytic activity. Journal of Agricultural and
Food Chemistry 16, 1031–1035.
Gurfinkel, D.M., Rao, A.V., 2002. Determination of saponins in legumes by direct
densitometry. Journal of Agricultural and Food Chemistry 50, 426–430.
Gurfinkel, D.M., Rao, A.V., 2003. Soyasaponins: the relationship between chemical
structure and colon anticarcinogenic activity. Nutrition and Cancer 47, 24–33.
Heng, L., Vincken, J.P., Hoppe, K., Koningsveld, G.A., Decroos, K., Gruppen, H., Boekel,
M.A.J.S., Voragen, A.G.J., 2006. Stability of pea DDMP saponin and the
mechanism of its decomposition. Food Chemistry 99, 326–334.
Hubert, J., Berger, M., Daydé, J., 2005. Use of simplified HPLC–UV analysis for
soyasaponins B determination: study of saponin and isoflavone variability in
65
soybean cultivars and soy-based health food products. Journal of Agricultural
and Food Chemistry 2005 (53), 3923–3930.
Jeon, H.S., Sung, M.K., 2000. Soybean saponins inhibit the formation of DNA adducts
colon and liver cells. Journal of Nutrition 130, 687S.
Jood, S., Chauhan, B.M., Kappor, A.C., 1986. Saponin content of chickpea and black
gram: varietals differences and effects of processing and cooking methods.
Journal of Science Food Agriculture 37, 1121–1124.
Kataria, A., Chauhan, B., Gandhi, S., 1988. Effect of domestic processing and cooking
on the antinutrients of black gram. Food Chemistry 30, 149–156.
Kataria, A., Chauhan, B., Punia, D., 1989. Antinutrients and protein digestibility
(in vitro) of mungbean as affected by domestic processing. Food Chemistry 32,
9–17.
Khalil, A.H., El-Adawy, T.A., 1994. Isolation, identification and toxicity of saponin
from different legumes. Food Chemistry 50, 197–201.
Khokhar, S., Chauhan, B.M., 1986. Antinutritional factors in moth bean (Vigna
aconififolia): varietal differences and effects of methods of domestic processing
and cooking. Journal of Food Science 51, 591–594.
Lee, S., Simons, A.L., Murphy, P.A., Hendrich, S., 2005. Soyasaponins lowered plasma
cholesterol and increased fecal bile acid in female golden Syrian hamsters.
Experimental Biology and Medicine 230, 472–478.
Matsuura, H., 2001. Saponins in garlic as modifiers of the risk of cardiovascular
disease. Journal of Nutrition 131, 5s–1000s.
Oakenfull, D., Sidhu, G.S., 1990. Could saponins be a useful treatment for
hypercholesterolaemia. European Journal of Clinical Nutrition 44, 79–88.
Oda, K., Matsuda, H., Murakami, T., Katayama, S., Ohgitani, T., Yoshikawa, M., 2003.
Relationship between adjuvant activity and amphipathic structure of
soyasaponins. Vaccine 21, 2154–2160.
Oh, Y.J., Sung, M.K., 2001. Soybean saponins inhibit cell proliferation by suppressing
PKC activation and induce differentiation of HT-29 human colon
adenocarcinoma cells. Nutrition and Cancer 39, 132–138.
Rao, A.V., Sung, M.K., 1995. Saponins as anticarcinogens. American Institute of
Nutrition (Am. Inst. Nutr.) 125, 717–724.
Sharma, A., Sehgal, S., 1992. Effect of processing and cooking on the antinutritional
factors of faba bean (Vicia faba). Food Chemistry 43, 383–385.
Shi, J., Arunasalam, K., Yeung, D., Kakuda, Y., Mittal, G., Jiang, Y., 2004. Saponins from
edible legumes: chemistry, processing, and health benefits. Journal of Medicinal
Food 7, 67–68.
Shimelis, E.A., Rakshit, S.K., 2007. Effect of processing on antinutrients and in vitro
protein digestibility of kidney bean (Phaseolus vulgaris L.) varieties grown in
east Africa. Food Chemistry 103, 161–172.
Shiraiwa, M., Harada, K., Okubo, K., 1991. Composition and structure of ‘‘group B
saponin” in soybean seed. Agriculture Biological Chemistry 55, 911–917.
Tarade, K.M., Singhal, R.S., Jayram, R.V., Pandit, A.B., 2006. Kinetics of degradation of
saponins in soybean flour (Glycine max) during food processing. Journal of Food
Engineering 76, 440–445.
Upadhyay, J.K., Garcia, V.V., 1988. Effect of soaking and cooking on reduction of
oligosaccharides of cowpea (Vigna unguichlata (L.) Walp.). Philippines Journal of
Food Science and Technology 12, 21–28.
Van der Poel, A.F.B., 1990. Effects of processing on antinutritional factors and
protein nutritional value of dry beans (Phaseolus vulgaris L.). Animal Feed
Science and Technology 29, 179–208.
Wang, Y., Zhang, Y., Zhu, Z., Zhu, S., Li, Y., Li, M., Yu, B., 2007. Exploration of the
correlation between the structure, hemolytic activity, and cytotoxicity of
steroid saponins. Bioorganic and Medicinal Chemistry 15, 2528–2532.