jfq_347 742..

757

THE EFFECTS OF DIFFERENT DEBITTERING METHODS ON

THE PRODUCTION OF LUPIN BEAN SNACK FROM BITTER
LUPINUS ALBUS L. SEEDS

MUSTAFA ERBAS1

Department of Food Engineering, Faculty of Engineering

Akdeniz University
07070 Antalya, Turkey

Received for Publication January 21, 2009

Accepted for Publication February 23, 2010

ABSTRACT

The aim of this study was to remove the alkaloids from bitter lupin
(Lupinus albus L.) seeds by different debittering methods and to produce a
lupin bean snack. During production of the snack, the total alkaloid content of
seeds decreased significantly (P ⱕ 0.01) from 14.4 g/kg to a level that was
undetectable. A major alkaloid in lupin seeds was identified as lupanine. It was
present at a concentration of 12.513 g/kg, which constituted 87% of the total
alkaloid content. The other alkaloids that were identified were sparteine,
albine, a-isolupanine and multiflorine. The concentrations of the alkaloids
decreased gradually during the debittering process. The values for dry matter,
protein, lipid, starch and ash content of seeds changed in a statistically
significant manner (P ⱕ 0.05, P ⱕ 0.01) during production of the snack.
Aqueous debittering at room temperature was found to be a more favorable
process because it resulted in more acceptable sensorial properties than
alkaline and thermal debittering methods.

PRACTICAL APPLICATIONS

Bitter lupin seeds (Lupinus albus L.) cannot be consumed directly

because they naturally contain high amounts of toxic alkaloids, which cause
bitterness, despite their high protein content. The aim of this study is to remove
bitterness from lupin seeds with different debittering methods and to produce
a lupin bean snack.

1
Corresponding author. TEL: +90-242-3106575; FAX: +90-242-2274564; EMAIL: erbas@
akdeniz.edu.tr

Journal of Food Quality 33 (2010) 742–757.

742 DOI: 10.1111/j.1745-4557.2010.00347.x
© 2010 Wiley Periodicals, Inc.
 SNACK FROM BITTER LUPINUS ALBUS L. SEEDS 743

INTRODUCTION

The seeds of members of the genus in the family Fabaceae constitute an

important part of the diet of the human population. They provide approxi-
mately 20% of the total protein that is consumed worldwide (ANZFA 2001;
Jimenez-Martinez et al. 2001). The major cultivated species of Lupinus
(lupins) genus belonging to Fabaceae are L. albus L. (white lupin), L. angus-
tifolius L. (blue lupin), L. luteus L. (yellow lupin) and L. mutabilis (Andean
lupin or tarwi). The first three of these are commonly cultivated in Mediter-
ranean countries and Australia, while L. mutabilis is cultivated in South
America (Allen 1998; Petterson 1998; Mulayim et al. 2002). L. albus seeds
have a high protein content ranging from 33 to 47%, according to genotype
and location (Huyghe 1997). This protein is rich in essential amino acids,
especially lysine. In addition, they can contain more than 150 g/kg of lipid and
dietary fiber, but this value depends on genotype and location (Petterson 1998;
Erbas et al. 2005; Sujak et al. 2006; Uzun et al. 2007). However, the usage of
bitter lupin seeds has been limited by the presence of toxic and bitter quino-
lizidine alkaloids (ANZFA 2001; Jimenez-Martinez et al. 2001). The quino-
lizidine alkaloids result in moderate acute toxicity and the LD50 level of
lupanine, which is the most abundant of these alkaloids in lupin, is 0.41 g/kg
for oral intake in rats (ANZFA 2001; Sanchez et al. 2005).
Lupins can be divided into sweet lupins, which contain low levels of
alkaloids, and bitter lupins, which contain higher levels of alkaloids. Debitter-
ing, which is an ancient procedure, involves the elimination of anti-nutritional
factors to improve the nutritive value, and it is widely used to wash out the
bitter components of seeds. Due to the fact that the alkaloids of lupin are
water-soluble, the alkaloid level in bitter lupins (0.05–4 g/kg) can be easily
decreased to levels that are safe for human consumption, by boiling the seeds
and then soaking them in water (ANZFA 2001). However, this processing may
unfortunately remove a large proportion of the soluble proteins, minerals,
flavonoids, monosaccharides and sucrose from the seeds, in addition to the
undesirable bitter and toxic alkaloids and the flatulence-inducing oligosaccha-
rides. The debittered lupin seeds can be also referred to as sweet lupin when
the alkaloid content has been reduced to less than 0.2 g/kg, which is current
maximum permitted concentration and adequate for safe consumption
(ANZFA 2001). In addition, sweet lupins have been to grow sweet genetic
varieties with low alkaloid contents that range from 0.08 to 0.12 g/kg.
However, this has ecological disadvantages because bitterness is a dominant
genetic characteristic (Allen 1998; Reinhard et al. 2006).
Lupin seeds can be used as a source of supplementary protein and fiber in
existing or novel food products. Joray et al. (2007) have investigated the
development of coated snacks from L. albus seeds. Debittered lupin seeds or
744 M. ERBAS

flour can be used in baking, and in the production of pasta, emulsified meat and
a variety of other food products to increase their nutritional value and to
improve aroma and texture. Also, lupin can be a good choice for vegetarians as
regards protein abundance (Mohamed and Rayas-Duarte 1995; Lampart-
Szczapa et al. 1997; Papavergou et al. 1999; Pollards et al. 2002; Chapleau
and Lamballerie-Anton 2003; Johnson et al. 2003; Sanchez et al. 2005;
Lampart-Szczapa et al. 2006; Martinez-Villaluenga et al. 2006). In addition,
protein isolates can be produced from lupin seeds. The protein extracted from
lupin seeds has good nutritional and functional properties such as high emul-
sifying, water-binding and foaming capacities (Mohamed and Rayas-Duarte
1995; El-Adawy et al. 2001).
Lupin seeds have been used as a food source by the European, Mediter-
ranean, Middle Eastern and Andean people since ancient times, especially in
soups, stews, salads and snacks. In some European and Middle Eastern coun-
tries, pickled lupin seeds, which are also known as “lupini beans,” are com-
monly sold in brine, in a similar manner to olives, for human consumption
(Petterson 1998; Papavergou et al. 1999; Pollards et al. 2002). This product
can be consumed as a snack with or without the husk. White lupin seeds that
have been boiled and soaked in water until they have been debittered and then
salted are known as “tirmis” or “termiye” in Turkey. They are similar to the
lupini beans.
The objective of this study was to remove the bitter alkaloids from lupin
seeds by the use of different debittering methods, and to introduce production
of a lupin bean snack.

MATERIALS AND METHODS

Raw Material
The lupin seeds (L. albus L.) were cultivated in the Gebiz village
(37°08′N, 30°58′E), which is located in the province of Antalya, Turkey, in
2005, and obtained from a local producer. The material was derived from the
landrace of the region.

Debittering of Lupin Seeds and Production of a Lupin Bean Snack

The production process for the lupin bean snack consisted of cleaning,
boiling, debittering and salting stages. Extraneous material and immature and
damaged seeds were removed first. The cleaned seeds were boiled in water
(1:3, seeds : water) for 75 min to destroy thermolabile anti-nutritional factors,
such as trypsin inhibitors and to soften the seeds. The boiled lupin seeds were
then strained and divided into three portions, to which different debittering
 SNACK FROM BITTER LUPINUS ALBUS L. SEEDS 745

methods were applied. The first group was debittered with water at room
temperature (~25C), the second group was debittered with 0.5% NaHCO3
(sodium bicarbonate) solution at room temperature (~25C) and the third group
was debittered with hot water (65C). The first, second and third groups were
coded as N (normal), A (alkaline) and T (thermal), respectively. The lupin
seeds, during the debittering process, were covered fully with debittering
liquids and these steps were renewed subsequently in 12-h intervals for 144 h.
After debittering, the seeds were washed gently with water (10 s) and trans-
ferred to salt water (6%, NaCl) for 12 h. The brine was then removed to yield
the lupin bean snack. Samples were taken for subsequent analysis at each
stage, and each time the debittering liquid was changed. The samples were
stored at -18C for later analysis.

Physical Analysis of Seeds

The weight and volume of 1000 seeds were determined by measuring
scale and filled with water cylinder. The water activity of the seeds was
measured with a water activity meter (Testo 650, Lenzkirch, Germany). The
hardness test was performed by pressing horizontally on a seed, using a texture
analysis device (TA TX Plus; Stable Micro Systems, Surrey, UK) that was
equipped with an SMS5 cylinder probe, until the first crack formed. Seed color
(L*, a* and b* values) was analyzed by the CIELAB system using a CR-400
chromameter (Konica Minolta, Osaka, Japan) that was equipped with a mea-
suring head and DP-400 data processor. The L* value represents the light–dark
spectrum with a range of 0 (black) to 100 (white), the a* value represents the
green–red spectrum with a range of -60 (green) to +60 (red) and the b* value
represents the blue–yellow spectrum with a range of -60 (blue) to +60
(yellow). The chromameter was calibrated using a white ceramic calibration
tile. Ten independent samples were analyzed and two readings from each
sample were recorded. The color differences between the raw lupin seeds and
the lupin bean snack were calculated using the following equation:
DE = (Da2 + Db2 + DL2)0.5, where DL, Da and Db are the differences in color
values between the raw and processed seeds (Ozdemir and Devres 2000).

Proximate Analysis of Seeds

The amount of lost dry matter during the debittering process was calcu-
lated from the weight difference between 100 dried seeds that were sampled
before and after snack production. Prior to the chemical analyses, the seeds
were dried and milled into a fine powder. The amount of dry matter was
determined by drying the samples at 105C to a constant weight (44–01 AACC
2000). The nitrogen content was determined by the Kjeldahl method and
multiplied by a factor of 6.25 (Lampart-Szczapa et al. 2003) to determine the
746 M. ERBAS

crude protein content (46–12 AACC 2000). The ash content was determined
by incineration of the sample at 925 °C to constant weight (08–01 AACC
2000). The lipid content was determined by the Soxhlet method (30–25 AACC
2000). The crude fiber content of the samples was determined, after the lipid
had been removed, by acid–base digestion to remove the starches and proteins
(32–10 AACC 2000). The starch content was determined by the polarimetric
method (TSE 2000). The pH values were measured using a pH meter (WTW
537, Weilheim, Germany) in 5 g samples, after homogenization of the seeds in
45 mL distilled water. The titratable acidity was determined by titration with
0.1 M NaOH up to Ph 8.1, and expressed in terms of sulfuric acid. All the
measurements were expressed as a function of dry weight.

Measurement of Alkaloids
The dried and milled seeds (0.5 g) were weighed in a tube (PPCO,
Nalgene, Rochester, NY), and homogenized in 10 mL petroleum ether for
1 min at 3,000 rpm using an Ultra-Turrax T-25 homogenizer (IKA Labortech-
nik, Staufen, Germany). They were then incubated at 40C for 30 min, centri-
fuged in a 3K30 centrifuge (Sigma Laborzentrifugen, Harz, Germany) at
4,500 ¥ g for 5 min, and the supernatant was removed. This process was
repeated twice, with the exception of the incubation at 40C, to remove the
lipids. The defatted and dried samples were homogenized in 5 mL trichloro-
acetic acid solution (0.3 M) using an Ultra-Turrax T-25 homogenizer, centri-
fuged at 4,500 ¥ g for 15 min and the supernatant collected in a beaker. This
process was also repeated twice to extract the alkaloids from the solid phase.
The supernatants that had been collected in the beaker were made alkaline by
adding 1 mL 10 M NaOH, followed by gentle agitation and resting. The
alkaloids were extracted from the alkalized supernatant with 5 mL dichlo-
romethane four times. The combined 20 mL of dichloromethane extract was
evaporated in a tube at 40C, and the resulting alkaloids were dissolved in 1 mL
methanol. The volume of 1 mL of each sample was injected in parallel into a
Fisons HRGC Mega 2 Series gas chromatograph (Milan, Italy) that was
equipped with a capillary column (HP-1, 19091Z-212, Agilent, Santa Clara,
CA; 25 m length, 0.32 mm i.d., 1.05 mm film thickness). The injector block
and flame ionization detector (FID) were maintained at 250C and 300C,
respectively. The oven temperature was increased gradually from 150C to
235C at 5C/min (Muzquiz et al. 1994; Muzquiz 2000; Sanchez et al. 2005).
The pressure of the carrier gas (helium) was 150 kPa, and the pressures of the
hydrogen and dry air that were used in the FID were 50 and 90 kPa, respec-
tively. The peaks were identified using sparteine as a standard (MP Biomedi-
cals, Solon, OH), and retention times were compared. To measure the degree
of recovery, sparteine was added into some samples and extracted using the
 SNACK FROM BITTER LUPINUS ALBUS L. SEEDS 747

same method. The recovery was higher than 95%. The standard calibration
curve was prepared using sparteine in methanol. The response was linear over
the range 0.0000091–0.091 g/L and the regression coefficient was 0.99. The
results were calculated as a function of dry weight.

Sensory Evaluation
The lupin bean snacks that had been produced by the different debittering
methods were subjected to sensory analysis. The snacks were placed on white
plastic plates. They were coded and served to panelists at random on a white,
light bench at 3:00 p.m. The snacks were evaluated by six trained panelists
(three female, three male; 24–33 years old) who were familiar with the char-
acteristics of this type of snack, and were studying as research assistants at the
Department of Food Engineering of Akdeniz University, Turkey. The panelists
participated in a group discussion to establish terms that described the char-
acteristics of the lupin bean snack. The descriptive terms those were selected
as appearance, color, smell, texture, taste, bitterness and overall. The accept-
ability of the lupin snack was scored by the panelists on a 5-point scale
(1 = disliked extremely, 5 = liked extremely). Sensory scores obtained from
the six trained panelists were averaged and analyzed by analysis of variance
(ANOVA) with respect to debittering methods.

Statistical Analysis
The physical and chemical analyses were performed on samples from the
four stages of production of the lupin bean snack, which corresponded to raw,
boiled, debittered seeds and the final product (snack). However, analysis of the
alkaloids was performed for each stage, as well as for each change of the
debittering liquid because alkaloids have restricted effects in the consumption
of lupin. In the research, production of the lupin bean snack were realized in
two replicates, and analyses were duplicated. The data were analyzed by
ANOVA using the SAS statistical software package (v.7.00, SAS Institute Inc.,
Cary, NC) to compare the means with respect to debittering time and method.
The Duncan’s multiple range test was used to determine significant differences
at the 5% level. Results are given as the mean ⫾ standard deviation.

RESULTS AND DISCUSSION

Changes in Properties of Seeds during Processing of Lupin Bean Snack

The weights of 1,000 raw and 1,000 processed seeds were 319 and 702 g,
respectively. The increases in the weight and volume of the seeds during the
748 M. ERBAS

production of the snack were 120 and 60%, respectively. These increases were
due to the high hydration capacity of the lupin proteins (Chapleau and
Lamballerie-Anton 2003; Lampart-Szczapa et al. 2006). The gains in the
weight and volume of the seeds showed a similar trend for all the debittering
methods. It has been reported that the seeds absorb an amount of water that is
approximately equal to twice their original weight during the debittering
process (Jimenez-Martinez et al. 2001). Solomon (2007) has reported similar
findings with respect to the weight gain of lupin seeds that had been soaked in
water at different temperatures.
The loss of dry matter from the seeds during the process was ~260 g/kg.
This loss was a result of the loss of fiber (~60 g/kg), which occurred because
of separation of the softened husks and soluble material (~200 g/kg) during the
debittering process. The detached husks could be observed as small particles in
all the changes of extraction liquid during debittering. In addition, the extrac-
tion liquids became yellowish in color because of the presence of pigments.
Jimenez-Martinez et al. (2001) have reported that during the process of deb-
ittering, 120–270 g/kg of solids were lost, which depended on the specific
treatment that was used. The other changes in the physical and chemical
properties of the seeds, which depended on the processing stage and time, are
shown in Table 1. All the physical and chemical properties of the seeds were
affected significantly (P ⱕ 0.05, P ⱕ 0.01) by the processing stage and time.
The greatest changes in the properties of the seeds occurred during the debit-
tering stage. Dry matter was decreased by the introduction of water into the
seeds and the loss of soluble components and detached husks. Therefore, the
water activity of the seeds increased, whereas the hardness decreased. During
the debittering process, the components of the seeds were modified at different
rates. The protein and starch contents of seeds increased by 67 and 18 g/kg,
respectively. These increases may have been caused by the fact that oligosac-
charides, minerals, alkaloids, flavonoids and fiber were removed when the
debittering liquid was changed. The crude fiber content decreased by ~60 g/kg
during the debittering process because of the loss of the husks. The ash content
of the seeds decreased as a result of the decrease in crude fiber content and the
removal of minerals by the extraction process. The acidity of the seeds
decreased because of the loss of soluble material. The lipid content increased
by ~10 g/kg in the boiled seeds as compared to the raw seeds. This was
proportional the result of the loss of water-soluble solids during boiling. The
common decrease in lipid content that was observed during the snack produc-
tion may have been caused by removal of the lipid-rich embryo together with
the detached husk.
The production process caused a difference in color (DE) of 16 units
when the raw and processed seeds were compared. The color values (L*, a*
and b*) of the seeds changed from cream (71.3, 5.9 and 17.1; raw seeds) to
 SNACK FROM BITTER LUPINUS ALBUS L. SEEDS 749

TABLE 1.
THE PHYSICAL AND CHEMICAL PROPERTIES OF THE LUPIN SEEDS CHANGES DURING
THE DIFFERENT STAGES OF PRODUCTION OF THE LUPIN BEAN SNACK†

Properties Processing stage and time (h) Sign.

Raw (0) Boiled (1) Debittered (144) Snack (156)

Dry matter 896a ⫾ 2.4 411b ⫾ 4.5 291d ⫾ 6.9 311c ⫾ 7.9 **
Protein 413d ⫾ 1.2 451c ⫾ 4.5 516a ⫾ 9.8 480b ⫾ 10.0 **
Crude fiber 146a ⫾ 1.9 138a ⫾ 2.1 93.0b ⫾ 12.5 87.7b ⫾ 11.8 **
Lipid 98.7b ⫾ 0.2 109a ⫾ 0.2 87.2c ⫾ 4.2 79.4d ⫾ 1.7 **
Starch 27.7b ⫾ 1.3 48.0a ⫾ 0.2 47.9a ⫾ 0.9 46.1a ⫾ 0.7 **
Ash 25.7b ⫾ 0.6 22.3c ⫾ 0.0 23.3c ⫾ 1.5 31.3a ⫾ 0.5 **
Total alkaloids 14.4a ⫾ 0.2 6.2b ⫾ 0.2 0.01c ⫾ 0.0 LOQ **
Acidity (as H2SO4) 10.9a ⫾ 0.2 8.8b ⫾ 0.2 2.0c ⫾ 0.7 2.3c ⫾ 0.4 **
pH 5.40b ⫾ 0.01 5.52b ⫾ 0.02 6.47a ⫾ 0.23 6.29a ⫾ 0.14 **
Water activity 0.57c ⫾ 0.01 0.92b ⫾ 0.00 0.96a ⫾ 0.02 0.94ab ⫾ 0.01 **
Hardness (kg) ⱖ60a ⫾ 0.00 3.69b ⫾ 0.01 2.93c ⫾ 0.34 3.21c ⫾ 0.27 **
Color difference, DE – 12.5b ⫾ 0.64 14.8ab ⫾ 2.15 16.0a ⫾ 1.64 *

† Properties are given as the mean ⫾ standard deviation. The means are represent average for three
treatments (N, A and T). Chemical properties are expressed as a function of dry weight (g/kg). LOQ
represents the limit of quantitation.
Sign: Statistical significance: * P ⱕ 0.05, ** P ⱕ 0.01, n = 6 (two replicates for each of three debit-
tering methods).
Superscript letters beside the mean values denote values in the same line that are significantly different
by the Duncan’s multiple range test (P < 0.05).

golden yellow (63.2, 8.6 and 32.9; processed seeds) during snack production,
when water at room temperature was used. Lupin seeds contain high levels of
carotenoids and zeaxanthin, which give the cotyledon its bright yellow color
(El-Difrawi and Hudson 1979). The color changes may be a result of the
Maillard reaction, in addition to the loss of flavonoid pigments, which have
been shown to be present in lupin (Allen 1998).
In the raw seeds, the total concentration of alkaloids was 14.4 g/kg, and
this was reduced to below the detection limit (0.0000091 g/L) by the debitter-
ing process. It has been reported that the total alkaloid content of Turkish bitter
landraces of L. albus is 19.1 g/kg (Muzquiz et al. 1994). The changes in the
levels of seed components that occurred during debittering in this study were
similar to those that were described by other researchers (Jimenez-Martinez
et al. 2001; Torres et al. 2005). The protein content of the debittered lupin
seeds was determined to be 480 g/kg, and it was similar to other lupin species
point of view protein abundant. Furthermore, the crude fiber and lipid contents
were similar to those found in the seeds of other lupin species (Jimenez-
Martinez et al. 2001). Sujak et al. (2006) have reported that the protein, lipid,
crude fiber and ash contents of L. albus seeds are 363, 115, 144 and 34 g/kg,
750 M. ERBAS

respectively. Lupin seeds are low in starch (23 g/kg), whereas the seeds of
other common legumes have high starch content (Cerning-Beroad and Filiatre
1976). Therefore, the debittered and defatted lupin seeds may be a potential
source of material for the production of dietetic foods because they have a high
protein and dietary fiber content and low starch content as compared with other
legumes and cereals. Also, lupin seeds could be used to enrich different types
of food products with respect to protein and dietary fiber.

Changes in Alkaloid Content of Seeds during Production of Lupin

Bean Snack
The alkaloid content of the seeds showed significant differences
(P ⱕ 0.01) between the different stages of production of the snack. The
changes are shown in Table 2. Five components were identified during gas
chromatography that, together, exceeded 95% of the total alkaloid content. The
major alkaloid in the lupin seeds was identified as lupanine with a concentra-
tion of 12.513 g/kg, which constituted 87% of the total alkaloid content.
Sparteine, albine, a-isolupanine and multiflorine were the other alkaloids that
were identified in this study. The alkaloid content decreased gradually during
the debittering process; sparteine was eliminated first, after 12 h in boiling
water, and lupanine was eliminated last, after 120 h. However, the debittered
seeds were soaked in water for an additional 24 h in order to ensure that they
were safe to consume. The amount of a-isolupanine did not show a steady
decrease; rather, its level also increased at certain stages during the production
process, which could have been caused by isomer transformations among the
alkaloids. Cuadra et al. (1994) reported that alkaloids underwent isomeriza-
tion. In lupin seeds, lupanine is the precursor of the other alkaloids (Sanchez
et al. 2005).
The results of this study with respect to the alkaloids are in agreement
with the findings of previous studies (Cuadra et al. 1994; Sanchez et al. 2005).
Muzquiz et al. (1994) examined 29 bitter ecotypes of L. albus, and reported
that lupanine was the main alkaloid (0.5–18.8 g/kg), and albine and multiflo-
rine were the other major alkaloids. Albine, a-isolupanine, lupanine, multiflo-
rine and 13-hydroxylupanine were found in all ecotypes, and their levels in the
Turkish ecotype were 0.2, 0.1, 11.2, 0.9 and 0.6 g/kg, respectively. Sanchez
et al. (2005) reported that the lupanine level in L. albus was 13.6 g/kg.
Sparteine has been found in some ecotypes of L. albus that are cultivated in
Europe (Petterson 1998).

Comparison of Different Debittering Methods

Some physical and chemical properties of the seeds are shown in Table 3
to allow comparison of the debittering methods. The values for protein, crude
 TABLE 2.
ALKALOID CONTENT OF THE LUPIN SEEDS CHANGES DURING THE DIFFERENT STAGES OF PROCESSING OF THE LUPIN
BEAN SNACK†

Time (h) and stage Sparteine Albine a-isolupanine Lupanine Multiflorine

a a a a
0 (Raw) 0.018 ⫾ 0.007 0.091 ⫾ 0.027 0.150 ⫾ 0.015 12.513 ⫾ 0.064 1.603a ⫾ 0.087
1 (Boiled) 0.011b ⫾ 0.001 0.048ab ⫾ 0.014 0.093bcd ⫾ 0.017 5.672b ⫾ 0.028 0.395b ⫾ 0.132
12 LOQ 0.034ab ⫾ 0.014 0.045cde ⫾ 0.021 3.117c ⫾ 0.702 0.159bc ⫾ 0.067
24 0.016ab ⫾ 0.007 0.018e ⫾ 0.003 0.869d ⫾ 0.092 0.052c ⫾ 0.013
36 0.007b ⫾ 0.002 0.037ed ⫾ 0.015 0.521d ⫾ 0.126 0.050c ⫾ 0.019
48 0.007b ⫾ 0.003 0.077bcd ⫾ 0.024 0.261d ⫾ 0.058 LOQ
60 LOQ 0.108ab ⫾ 0.035 0.165d ⫾ 0.036
72 0.059cde ⫾ 0.019 0.052d ⫾ 0.017
84 0.099abc ⫾ 0.025 0.034d ⫾ 0.009
96 0.043cde ⫾ 0.010 0.014d ⫾ 0.005
108 0.007e ⫾ 0.003 0.007d ⫾ 0.002
120 LOQ 0.008d ⫾ 0.002
132 LOQ
144 (Debittered) LOQ
156 (Snack) LOQ
Sign. ** ** ** ** **
SNACK FROM BITTER LUPINUS ALBUS L. SEEDS

Sign: Statistical significance: ** P ⱕ 0.01, n = 6 (two replicates for each of three debittering methods).
† Alkaloid content is given as the mean ⫾ standard deviation as a function of dry weight (g/kg). The means are represent average for three treatments
(N, A and T). LOQ represents the limit of quantitation.
Superscript letters beside the mean values denote values in the same column that are significantly different by the Duncan’s multiple range test (P < 0.05).
751
752 M. ERBAS

TABLE 3.
SOME PHYSICAL AND CHEMICAL PROPERTIES OF THE LUPIN SEEDS TREATED BY
THE DIFFERENT DEBITTERING METHODS†

Properties Debittering methods Sign.

N A T

Dry matter 354a ⫾ 27.5 353a ⫾ 27.6 358a ⫾ 27.4 –

Protein 469a ⫾ 15.4 475a ⫾ 18.2 451b ⫾ 10.5 *
Crude fiber 112b ⫾ 11.1 102c ⫾ 15.5 134a ⫾ 5.8 **
Lipid 96.6a ⫾ 4.4 91.2b ⫾ 5.0 92.9b ⫾ 4.4 *
Starch 42.8a ⫾ 3.4 41.7a ⫾ 3.2 42.7a ⫾ 3.3 –
Ash 25.5a ⫾ 1.5 26.2a ⫾ 1.5 25.2a ⫾ 1.3 –
Total alkaloid 2.5a ⫾ 1.0 2.8a ⫾ 1.1 2.4a ⫾ 1.0 –
Acidity (as H2SO4) 6.7a ⫾ 1.2 5.9ab ⫾ 1.5 5.4b ⫾ 1.7 *
pH 5.69b ⫾ 0.11 6.08a ⫾ 0.26 5.98a ⫾ 0.21 *
Water activity 0.84a ⫾ 0.06 0.84a ⫾ 0.06 0.85a ⫾ 0.06 –
Hardness (kg) 17.4a ⫾ 9.30 17.1a ⫾ 9.35 17.8a ⫾ 9.20 –
Color difference, DE 15.9a ⫾ 1.25 16.8a ⫾ 1.50 10.6b ⫾ 0.90 **

Sign: Statistical significance: * P ⱕ 0.05, ** P ⱕ 0.01, –: P ⱖ 0.05, n = 8 (two replicates for each of
the four processing stages (raw, boiled, debittered seeds and snack).
† Properties are given as the mean ⫾ standard deviation. Chemical properties are expressed as a
function of dry weight (g/kg).
Superscript letters beside the mean values denote values in the same line that are significantly different
by the Duncan’s multiple range test (P < 0.05).
N: Normal aqueous debittering at room temperature. A: Alkaline (0.06 M NaHCO3) debittering at room
temperature. T: Thermal aqueous debittering at 65C.

fiber and lipid content, acidity, pH, and color differences of the seeds did show
significant differences (P ⱕ 0.01, P ⱕ 0.05), whereas between the different
debittering methods, the values for dry matter, starch, ash, water activity and
hardness did not differ significantly (P ⱖ 0.05). There were no significant
(P ⱖ 0.05) differences between the total alkaloids content of the lupin seeds
produced by different debittering treatments.
The acidity and pH values were different in the A group because of the
alkaline treatments, and the color difference was different in the T group
because of the high temperature treatments at 65C. The color of seeds in the T
group was darker than the others. This darkness might be resourced that
Maillard reaction, non-enzymatic browning reaction between reducing sugar
and free amino groups, was encouraged by high temperature. The protein
content was low in the T group, it is possible that this differences resourced
from loss water-soluble nitrogen contain compounds due to increase the solu-
bility and cellular permeability in high temperature. The crude fiber contents
were the lowest in the A group that is treated with sodium bicarbonate because
of the alkali effect. The lowness might be due to fact that alkaline effect
 SNACK FROM BITTER LUPINUS ALBUS L. SEEDS 753

detached husk of lupin seed. Saxena et al. (1990) has reported sodium bicar-
bonate solutions have been used for dehulling of pulses. The lipid contents in
the A and T treatments were determined lower than the N. The decrease in lipid
content in the A could be a result of the removal of the lipid-rich embryo
together with the detached husk. This decrease in T could be arose from the
temperature’s lipid-melting effect. All of the lost components mentioned
earlier were eliminated during the changing of debittering water.

Sensory Evaluation of Lupin Snacks

All of the sensorial properties of the lupin snack were affected signifi-
cantly (P ⱕ 0.01, P ⱕ 0.05) by the debittering methods (Table 4). The snack
that was produced by debittering with water at room temperature (N) showed
the highest degree of preference because it had a golden yellow color, and a
good taste and texture, although it is same statistic group with the A that was
produced alkaline debittering. The panelists found the snack that was prepared
by thermal treatment (T) to be unacceptable. All of the sensorial properties of
the T snack were worse than those of the others (N and A). This may be a result

TABLE 4.
SENSORIAL PROPERTIES OF LUPIN BEAN SNACKS
HAVE BEEN PRODUCED BY THE DIFFERENT
DEBITTERING METHODS†

Properties Debittering methods Sign.

N A T

Appearance 4.6a ⫾ 0.4 3.9a ⫾ 0.1 1.8b ⫾ 0.4 *

Color 4.2a ⫾ 0.2 4.4a ⫾ 0.2 1.4b ⫾ 0.2 **
Smell 4.3a ⫾ 0.3 4.1a ⫾ 0.3 2.6b ⫾ 0.6 *
Texture 4.5a ⫾ 0.4 4.3a ⫾ 0.6 2.3b ⫾ 0.1 *
Taste 4.6a ⫾ 0.1 4.0a ⫾ 0.3 2.0b ⫾ 0.4 **
Bitterness 4.3a ⫾ 0.0 4.0a ⫾ 0.4 2.7b ⫾ 0.5 *
Overall 4.6a ⫾ 0.3 3.9a ⫾ 0.2 2.1b ⫾ 0.6 *
Acceptance yes yes no

Sign: Statistical significance: * P ⱕ 0.05, ** P ⱕ 0.01, n = 2 (two

replicates, snack).
† Properties are given as the mean ⫾ standard deviation in a range
from 1 to 5.
Superscript letters beside the mean values denote the values in the
same line that are significantly different by the Duncan’s multiple
range test (P < 0.05).
N: Normal aqueous debittering at room temperature. A: Alkaline
(0.06 M NaHCO3) debittering at room temperature. T: Thermal
aqueous debittering at 65C.
754 M. ERBAS

of the browning reaction, auto-oxidation and thermal degradation of the seed

components, which occur during debittering at high temperature. The T snack
had an unpleasant smell and taste, and was dark in color. The color of food is
one of the most important attributes for its acceptance. Therefore, the color of
the lupin bean snack was a useful and practical attribute for the comparison of
the debittering methods and snack quality. None of the panelists stated any
bitterness in the snacks, although T got a lower bitterness score. It might have
sourced that bitterness all of snacks have acceptable level.

CONCLUSIONS

In conclusion, the debittering methods that were used in this study

enabled the successful removal of alkaloids from lupin seeds. The normal
aqueous debittering method, which used water at room temperature, was found
to be the superior process because it was more straightforward to perform and
resulted in more acceptable sensorial properties than the other two methods.
This method could be used to produce a commercial lupin bean snack. The
absence of a bitter taste and the presence of a golden yellow color (L*, a* and
b*; 63.2, 8.6 and 32.9, respectively) were considered to be important quality
criteria. The lupin bean snack provides a suitable and safe means of consump-
tion of lupin seeds and a valuable nutritional source, especially as a dietetic
food, because of its high protein and dietary fiber content and its low starch
content.

ACKNOWLEDGMENTS

This research was supported financially by the Akdeniz University

Research Fund (Project Number 2004.01.0104.005).

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