JOURNAL OF FOOD COMPOSITION AND ANALYSIS (2000) 13, 193}199

doi:10.1006/jfca.1999.0887
Available online at http://www.idealibrary.com on

ORIGINAL ARTICLE

Chemical Composition and Antinutrient Content of three

Lupinus Species from Jalisco, Mexico
M. A. Ruiz-LoH pez*, P. M. GarcmH a-LoH pez, H. Castan eda-Vazquez*, N. J. F. Zamora*,
P. GarzoH n-De la Mora-, J. Ban uelos Pineda*, C. Burbano?, M. M. Pedrosa?,
C. Cuadrado?, and M. Muzquiz?
*Centro Universitario de Ciencias Biolo& gicas y Agropecuarias, Universidad de Guadalajara, Las Agujas,
Nextipac, Zapopan, Jal. Km 15.5, Carretera Guadalajara-Nogales, Mexico; -Laboratorio de Bioqun& mica,
Depto. de Fisiologn& a, Centro Universitario de Ciencias de la Salud, Guadalajara, Jalisco, Mexico; ?Area de
Tecnologn& a de Alimentos, CIT-INIA, Aptdo. 8111,28080 Madrid, Spain

Received March 5, 1999, and in revised form January 24, 2000

In this study, the proximal chemical analysis and contents of antinutritional factors (lectins,
antitrypsin activity, cyanogenic glycosides, alkaloids, phytates and a-galactosides) of ¸upinus
exaltatus, ¸. re-exus, and ¸. mexicanus seeds were determined. The seeds of these lupins
comprised 384.1, 388.0, 367.0 g/kg protein, which contained all the essential amino acids for
human beings except glutamine and asparagine in their seed protein. Only ¸. exaltatus was free
from lectins. Trypsin inhibitor concentrations ranged from 1.12 to 2.05 TIU/mg. Cyanogenic
glycosides were present at low concentrations in the studied lupins. Total alkaloid content
ranged from 14.0 to 44.0 g/kg and phytate content ranged from 11.1 to 1.856 g/kg. The main
a-galactosides found in seeds were ra$nose, stachyose, verbacose and ajucose. Stachyose was the
predominant sugar in the studied species. Therefore, these wild lupins could be considered
a good source of protein after a suitable reduction in the content of alkaloids.
 2000 Academic Press

Key Words: wild lupins; proximal composition; antinutritional factors.

INTRODUCTION

In the past, Greeks, Romans, Egyptians and Andean people cultivated lupins for soil
improvement and human consumption. Today, people of some Mediterranean coun-
tries such as Spain and Italy eat domesticated lupins as snack (Cowling et al., 1998;
Gladstones, 1970) and in Chile, lupin #our is consumed as an adjunct with bakery
products (Ballester et al., 1988). In Peru, wild lupins were consumed by the Incas
and are still consumed by the current population. In Mexico, although wild lupins
are abundant throughout the country, neither wild nor domesticated lupins have
been consumed or cultivated. However, they could represent an important source of
good-quality protein and could be utilized as oil source.
Wild lupins are known to contain toxic alkaloids and several antinutritive factors
and must be properly processed before consumption (Rahma and Narasinga, 1984;

To whom correspondence and reprint requests should be addressed: Tel./Fax: (3) 682-0003. E-mail:
mruiz@maiz.cucba.udg.mx

0889}1575/00/030193#07 $35.00/0  2000 Academic Press

194 RUIZ-LOD PEZ E¹ A¸.

Muzquiz et al., 1993a, b); however the properties and chemical composition of Mexi-
can wild lupins are poorly understood. Therefore, the objective of the present work
was to determine the proximate composition and the antinutrient content of ¸upinus
exaltatus, ¸. re-exus, and ¸. mexicanus from the state of Jalisco, Mexico, in order to
compare them with those observed in lupins from other areas of the world.

MATERIAL AND METHODS

Material

Whole plants and pods of ¸. exaltatus, ¸. re-exus, and ¸. mexicanus were collected
from July to September of 1997 at several locations of the state of Jalisco, Mexico.
After collection, plants were botanically identi"ed and properly archived at the
Botanical Institute of the Universidad de Guadalajara for future reference. Samples of
seeds of the collected species were removed and dried for 48 h at 603C. After drying,
the seeds were pooled and ground into a #our using a mortar and pestle and stored at
103C until all assays were performed.

Methods

Proximate composition. Protein, moisture, ether extract, ash and crude "ber contents
were determined as described in the AOAC methods. Defatted seed #our from each of
the lupins under study were hydrolyzed with 6 N HCl or 4 N NaOH (tryptophan) at
1053C during 24 h in order to hydrolyze proteins to further identify amino acids by
thin-layer chromatography (Sotelo and Lucas, 1988).

¸ectins. Hemaglutinins were semiquantitatively determined in accordance with a modi-

"ed method (Sotelo and Lucas, 1988) using de"brinated sheep and rabbit red-blood cells.
After de"brination, cells were treated with 0.1% trypsin solution in 0.85% NaCl for
30 min at 373C. Suspensions of trypsinized and 0.85% NaCl-washed red-blood cells
were added to a serial dilution of a lupin extract which was prepared by shaking 1 g of
raw lupin #our with 10 mL of l% NaCl solution for 2 h at 253C. Results were
expressed as the maximal dilution capable of producing a hemaglutination reaction.

¹rypsin inhibitors. The activity of trypsin inhibitors in lupin seeds was determined
following a method described elsewhere (Kakade et al., 1974). The antitrypsin activity
was expressed as TIU/mg of sample.

Cyanogenic glycosides. The cyanogenic glycosides were determined by the measure-

ment of released HCN from lupin #our (Tejada De Hernandez, 1990).

Alkaloids. Lupin alkaloids were quantitatively analyzed as described elsewhere (Baer

et al., 1979). Soybean was used as a negative control in this assay.

Phytic acid. Inositol phosphates were determined according to Cuadrado et al. (1996).
Lupin samples (0.5 g) were extracted with 5 mL of 0.5 M HCl for 3 h at room tem-
perature. The hydrochloric acid extract was puri"ed and concentrated using an
ion-exchange SAX column (Varian). The inositol tri (IP3), tetra (IP4), penta (IP5) and
hexaphosphate (IP6) were measured by ion-pair reverse-phase HPLC, using a C18
column (PRP-1, 5 lm, 150;4.1 mm, Hamilton). The mobile phase was 0.8% tetra-
butylammonium hydroxide (TBN-OH) in methanol : water (51.5 : 48.5).
 ANTINUTRIENTS AND WILD LUPINS 195

a-galactosides. The a-galactosides were extracted by use of ion exchange columns

(Muzquiz et al., 1992). HPLC was carried out on a Spherisorb-S5-NH2 column
(250;4.6 mm i.d.) with acetonitrile/water 65 : 35 (v/v) 1 mL/min, as the mobile phase.
Individual sugars were quanti"ed by comparison with external standards of sucrose,
ra$nose, stachyose, ajucose and verbacose.

RESULTS AND DISCUSSION

To date, ¸upinus albus (white lupin), ¸. angustifolius (narrow-leafed lupin), ¸. luteus

(yellow lupin) from Europe and ¸. mutabilis (tarwi or pearl lupin) native of South
America have been domesticated and cultivated throughout the world for animal and
human consumption (Gross and Baer, 1977; Manzanares and Estrella, 1985; Gross,
1982). Although there exist 80 wild ¸upinus species in Mexico, none have been used as
food for human consumption or as animal feed as soybean commonly is.

Proximate Composition

The proximate chemical analysis of wild ¸. exaltatus, ¸. re-exus and ¸. mexicanus

seeds is shown in Table 1. Crude protein (CP) content was similar for all lupin seeds,
which also showed a high content of dry matter (DM), and crude "ber (CF). The ash
(A) content of ¸. re-exus (72.2 g/kg) was much higher than for the other two species,
and the ether extract (EE) content of ¸. mexicanus (80.2 g/kg) slightly higher. The
nitrogen-free extract (NFE) content was similar in the three species and ranged
between 321.6 and 343.7 g/kg (Table 1). The proximate composition of wild ¸.
exaltatus, ¸. re-exus and ¸. mexicanus seeds is within the range of values that have
been reported for domesticated lupins as well as other wild lupins (NAS, 1979).
¸. exaltatus, ¸. re-exus, and ¸. mexicanus had EE values lower than those of seed of
¸. mutabilis and the CF values were similar to those of other lupins, both values being
higher than those reported for the majority of cultivated legumes except for soybean
(Giral et al., 1978). This indicates that lupins contain more structural carbohydrates
than the majority of cultivated legumes (Aguilera and Trier, 1978).
The three species of wild lupins studied contained lower crude protein (37}39%)
than that found in soybean (44%), ¸. luteus or ¸. mutabilis (41}43%) (Guillaume,
1923; Herrera, 1944; Muzquiz et al., 1989a). In addition, the following amino acids,
lysine, aspartic acid, hystidine, serine, alanine, threonine, glycine, glutamic acid,
leucine, isoleucine, methionine, cystine, tryptophane, phenylalanine, valine, proline,

TABLE 1
Chemical proximate analysis of seeds of ¸upinus exaltatus, ¸. re-exus and ¸. mexicanus (g/kg dry matter
basis, N"3)

¸. exaltatus ¸. re-exus ¸. mexicanus

Dry matter 948.1 828.4 818.0

Protein (N;6.25) 384.1 388.0 367.0
Ether extract 71.1 66.1 80.2
Crude "ber 185.2 152.0 168.0
Ash 38.0 72.2 41.1
NFE 321.6 321.7 343.7

Nitrogen-free extract.
196 RUIZ-LOD PEZ E¹ A¸.

tyrosine and arginine, were found in wild lupin seeds. Therefore, all essential amino
acids are present in these seeds in agreement with the information reported in the
literature (Muzquiz et al., 1989b; Haq, 1993).

Antinutritive Factors

Concentration of lectins, trypsin inhibitors, cyanogenic glycosides and total alkaloids

are shown in Table 2. When sheep erythrocytes were used, a positive lectin response
was found only for ¸. mexicanus. Conversely, only ¸. re-exus gave a positive lectin
response (at 3 titer) when rabbit erythrocytes were used. Seeds of all the lupins gave
positive results when tested for trypsin inhibitors. ¸. exaltatus, ¸. re-exus and ¸. mexi-
canus were found to contain 2.05, 1.37 and 1.12 TIU/mg of #our, respectively. The titer
of lectins and content of trypsin inhibitors in seeds of wild lupin were lower than those
found in ¸. mutabilis and potatoes (Solanum tuberosum) cultivars (Shoeneberger et al.,
1982; Sotelo et al., 1998).
A negligible cyanogenic glycosides content in seeds of wild lupin species has been
reported (Kingsbury, 1964). The low concentration ( ( 0.01 mg HCN per 100 g of
#our) found in three of the lupin species studied here is in agreement with this report.
Total alkaloid concentration, determined as lupanine, for ¸. exaltatus, ¸. re-exus
and ¸. mexicanus was 14.0, 21.0 and 44.0 g/kg, respectively. However, the alkaloid
values of the three Jalisco wild lupin species are higher than the concentration
permitted for safe human consumption which is 0.2 g/kg (Chango et al., 1993).
Intoxication and birth defects in human beings and cattle have been reported after

TABLE 2
Toxic and antinutritional factors present in seeds of ¸upinus exaltatus, ¸. re-exus and ¸. mexicanus (N"3)

¸. exaltatus ¸. re-exus ¸. mexicanus

Lectins
Sheep blood * * 3
Rabbit blood * 3 *
Trypsin inhibitors
(TUI/mg of #our) 2.05$0.36 1.37$0.36 1.12$0.68
Cyanogenic glycosides
(mg HCN per 100 g of
#our) (0.01 (0.01 (0.01
Alkaloids (g/kg) 14$0.06 21$0.50 44$0.10

Titer"maximal dilution at which agglutination is observed.

Determined as lupanine.
Means$standard deviation of three determinations for each sample.

TABLE 3
Inositol phosphate content (g/kg) in seeds of ¸upinus exaltatus. ¸. re-exus and ¸. mexicanus (N"3)

Sample IP3 IP4 IP5 IP6 Total

¸. exaltatus n.d. n.d. 0.35$0.02 11.34$0.30 11.69$0.32

¸. re-exus n.d. n.d. 0.35$0.06 10.74$0.49 11.09$0.55
¸. mexicanus 0.011$0.000 n.d. 0.45$0.04 16.13$0.36 16.63$0.37

n.d."Undetectable.
Means$standard deviation of three determinations for each sample.
 ANTINUTRIENTS AND WILD LUPINS 197

TABLE 4
Sucrose and a-galactosides content (g/kg) in seeds of ¸upinus exaltatus, ¸. re-exus and ¸. mexicanus

Total
Sample Sucrose Ra$nose Stachyose Verbacose Ajucose a-galactosides

¸. exaltatus 12.33$0.14 9.52$0.27 41.77$1.96 15.60$0.68 0.79$0.01 67.68$2.92

¸. re-exus 9.92$0.71 5.40$0.49 36.99$1.03 24.87$1.03 1.56$0.09 68.82$3.35
¸. mexicanus 26.10$0.86 13.62$0.47 34.45$1.47 3.96$0.17 0.41$0.14 52.44$2.25

Means$standard deviation of three determinations for each sample.

ingestion of either green seeds or the water used to debitter lupins (Keeler, 1989;
Luque and Gutierrez-Rave, 1991).
The total content of phytates ranged from 11.09 g/kg for ¸. re-exus to 16.63 g/kg
for ¸. mexicanus. IP5 and IP6 were found in all three lupin species while IP3 only
appeared in ¸. mexicanus (Table 3). The relative proportion of IP6 was around 97% of
the total inositol phosphates, while IP5 was only around 3%. IP4 was not detectable
in any of the three lupins. All three wild lupin species had a higher phytate content
than that reported for ¸. luteus or ¸. albus (De la Cuadra et al., 1994). However,
phytate concentration was similar to those reported for ¸. angustifolius (Dagnia et al.,
1992) and ¸. albus cv. Multolupa (Camacho et al., 1991).
The oligosaccharide composition of the studied ¸upinus species is shown in Table 4.
The a-galactosides found in these seeds were ra$nose, stachyose, verbacose and
ajucose. Stachyose was the predominant sugar in all species. Verbacose content was
highest in ¸. re-exus (24.87 g/kg) while ra$nose content was highest in ¸. mexicanus
(13.62 g/kg). The ajucose content ranged from 0.41 g/kg in ¸. mexicanus to 1.56 g/kg
in ¸. re-exus, representing only the 0.78}2.27% of whole-seed oligosaccharides. The
sucrose content ranged from 9.92 g/kg in ¸. re-exus to 26.1 g/kg in ¸. mexicanus. The
total a-galactoside content of ¸. re-exus and ¸. exaltatus was similar to that reported
for ¸. albus and ¸. luteus (Saini, 1989; De la Cuadra et al., 1994). ¸. mexicanus showed
lower a-galactoside values than ¸. re-exus and ¸. exaltatus, similar to those reported
elsewhere for ¸. albus cv. Multolupa (Camacho et al., 1991).
In conclusion, our results have demonstrated that ¸. mexicanus, ¸. exaltatus
and ¸. re-exus could be an important source of high-quality proteins and edible oil
with negligible trypsin inhibitors, lectin and cyanogenic compounds. However, soak-
ing or some processing will be necessary to eliminate or reduce the risk of alkaloid
toxicity.

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