Food Chemistry 161 (2014) 296–304

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Fatty acid, carotenoid and tocopherol compositions of 20 Canadian lentil

cultivars and synergistic contribution to antioxidant activities
Bing Zhang a,b, Zeyuan Deng a,⇑, Yao Tang b,c, Peter Chen b,d, Ronghua Liu b, D. Dan Ramdath b, Qiang Liu b,
Marta Hernandez b, Rong Tsao b,⇑
a
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China
b
Guelph Food Research Centre, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, ON N1G 5C9, Canada
c
Key Laboratory of Food Nutrition & Safety (Tianjin University of Science & Technology), Ministry of Education, Tianjin 300457, China
d
Department of Food Science, Ontario Agricultural College, University of Guelph, Guelph, ON N1G 2W1, Canada

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

Article history: Understanding the profile of lipophilic phytochemicals in lentils is necessary to better understand the
Received 24 January 2014 health benefits of lentils. The fatty acid, carotenoid and tocopherol compositions and antioxidant activ-
Received in revised form 28 March 2014 ities of the lipophilic extracts of 20 lentil cultivars (10 red and 10 green) were therefore examined. Lentils
Accepted 2 April 2014
contained 1.52–2.95% lipids, of which 77.5–81.7% were unsaturated essential fatty acids. Total tocophe-
Available online 13 April 2014
rols ranged from 37 to 64 lg/g DW, predominantly c-tocopherol (96–98% of the tocopherol content), fol-
lowed by d- and a-tocopherol. trans-Lutein was the primary and major carotenoid (64–78%) followed by
Keywords:
trans-zeaxanthin (5–13%). Carotenoids and tocopherols showed weak correlation with 2,2-diphenyl-1-
Lentils
Fatty acids
picrylhydrazyl (DPPH) activity (r = 0.4893 and 0.3259, respectively), but good correlation when combined
Tocopherols (r = 0.6688), suggesting they may act synergistically. Carotenoids were found to contribute the most to
Carotenoids the strong antioxidant activity measured by photochemiluminescence (PCL) assay. Results from this
Antioxidant activities study contribute to the development of lentil cultivars and related functional foods with increased health
benefits.
Crown Copyright Ó 2014 Published by Elsevier Ltd. All rights reserved.

1. Introduction
Legumes are the oldest crops cultivated by humans and are one
of the most important foodstuffs consumed in Europe, the Middle
East, Africa, and South Asia (Gumienna, Lasik, & Czarnecki, 2009).
In some developing countries, legumes are consumed as a basic
staple food with high quality and low cost protein alternatives to
animal proteins (Marathe, Rajalakshmi, Jamdar, & Sharma, 2011).
Among legumes, lentils (Lens culinaris) have been gaining
increasing attention for their nutritive value. They are considered
as a potential whole food source for people affected by micronutri-
ent malnutrition (Thavarajah, Thavarajah, Sarker, & Vandenberg,
2009). Canada is by far the world’s largest lentil producer with
an annual production of approximately 4 megatons accounting
for 25% of the total world lentil output (Thavarajah et al., 2009).
On average, while global pulse consumption is declining, the
annual consumption of lentils is steadily increasing (Zou, Chang,
⇑ Corresponding authors. Tel./fax: +86 791 88304402 (Z. Deng). Tel.: +1 226 217
8108; fax: +1 226 217 8183 (R. Tsao).
E-mail addresses: zeyuandeng@hotmail.com (Z. Deng), rong.cao@agr.gc.ca
(R. Tsao).
Gu, & Qian, 2011). Lentils are an excellent source of both macronu-
trients, micronutrients and phytochemicals (Dueñas, Hernández, &
Estrella, 2002). Phytochemicals of lentils may provide potential
health benefits for humans. Epidemiological and interventional
studies suggest that legume consumption, including lentils, is
inversely associated with the incidence of several chronic diseases,
for example, coronary heart disease, type II diabetes mellitus, car-
diovascular diseases, cancer and aging (Amarowicz & Pegg, 2008;
Villegas et al., 2008). However, the use of lentils in food products
has been limited in western countries, due to traditional eating
customs, lack of consumer understanding, processing techniques
and available diversified food products. Incorporation of lentils
into western diets has been highly recommended (Aguilera et al.,
2010; Han & Baik, 2008).
While many research groups have focused their studies on bio-
active components and attributed the potential health benefits to
the antioxidant activities of hydrophilic phytochemicals such as
phenolics in lentils, available information on the bioactive compo-
sitions and antioxidant activities of lipophilic compounds from len-
tils are lacking (Dueñas, Sun, Hernández, Estrella, & Spranger,
2003; Xu & Chang, 2010). There are a few studies on other legumes
and cereals in terms of their fatty acid profile and lipophilic

http://dx.doi.org/10.1016/j.foodchem.2014.04.014
0308-8146/Crown Copyright Ó 2014 Published by Elsevier Ltd. All rights reserved.
 B. Zhang et al. / Food Chemistry 161 (2014) 296–304
phytochemical contents, such as tocopherols and carotenoids
(Boschin & Arnoldi, 2011; Konopka, Czaplicki, & Rotkiewicz,
2006). In these legumes and cereals, lutein was found to be the
main carotenoid (Abdel-Aal, Young, Rabalski, Hucl, & Fregeau-
Reid, 2007). Tocopherols and carotenoids are strong antioxidants.
Lutein is a non-provitamin A oxygen-containing carotenoid or xan-
thophyll that has been associated with reduced incidence of age-
related macular degeneration, cataracts, cancer, and cardiovascular
disease (Olmedilla, Granado, Blanco, Vaquero, & Cajigal, 2001;
Osganian et al., 2003). It has also been found to play significant
roles in promoting the health of eyes and skin (Yao et al., 2013).
Understanding the profile of lipophilic phytochemicals in pulse
foods, particularly lentils is therefore necessary, in order to better
understand how these micronutrients in lentils contribute to
health benefits and how these bioactive components are retained
at maximal levels throughout the food processing chain.
The new trend in pulse consumption and its potential health
benefits require a closer look at the possible bioactive components
in grain legumes like lentils. There are many commercial lentil cul-
tivars, but the largest collection of such are found in Canada. In the
present study, we have selected 20 popular red and green Canadian
lentil cultivars, and conducted a comprehensive analysis of the
fatty acid profile, the contents of total and individual carotenoids,
tocopherols, as well as the antioxidant activities of the lipophilic
extracts that contained these compounds using two chemical-
based models, i.e. the photochemiluminescence (PCL) and 2,2-
diphenyl-1-picrylhydrazyl (DPPH) assays. Results from this study
will provide support for selecting lentil cultivars with improved
nutritional value, and lead to further study on how these bioactive
components contribute to human health.
2. Materials and methods
2.1. Plant materials
The 20 lentil cultivars and breeding lines used for this study
were received from Saskatchewan Pulse Growers (Canada) on Sep-
tember 26, 2012. The whole lentil samples were ground into fine
powder, and stored in sealed plastic bags at –4 °C prior to analysis.
The 20 tested lentils are categorised into 2 groups based on their
colours: 10 red lentils: Blaze, Redcliff, Maxim, Rouleau, Redbow,
Redberry, Impact, Imperial, Rosetown, and Dazil; 10 green lentils:
Imvincible, Greenland, Asterix, Imigreen, Impower, Improve, Sov-
ereign, Milestone, Eston and Plato.
2.2. Chemicals and reagents
All-trans-lutein and all-trans-zeaxanthin standards were pur-
chased from Indofine (Belle Mead, NJ); fluorescein, Trolox and
DPPH were obtained from Sigma (St. Louis, MO). All HPLC-grade
solvents, including methanol, methyl tert-butyl ether (MTBE), tet-
rahydrofuran, hexane and isopropanol were purchased from Cale-
don Laboratories (Georgetown, ON, Canada). All other chemical
reagents used were of analytical grade.
2.3. Colorimetric study
The colour of ground samples was measured at room tempera-
ture using a Minolta Chromometer (Chroma Meter CR-300; Minol-
ta Camera Co. Ltd., Osaka, Japan) according to the method of Li
et al. (2011). The chromometer consisted of an 8-mm-diameter
measuring area and diffuse illumination/viewing. The tristimulus
values of CIE L⁄, a⁄, b⁄ readings were calibrated against a standard
calibration white plate. CIE 1976 uniform colour space was taken
into account for the colorimetric analysis. Within the CIELAB uni-
297
form space, psychometric index of brightness L⁄ measures the
whiteness value of a colour and ranges from black at 0 to white
at 100, while chromaticity coordinate a⁄ is measured for redness
when positive and greenness when negative, and b⁄ indicates yel-
lowness when positive and blueness when negative. The values a⁄
and b⁄ were used to calculate the hue angle (H = arctan (b⁄/a⁄)) and
metric chroma value (C = (a⁄2 + b⁄2)1/2). The data of each measure-
ment are the average of triplicate measures on equidistant points
of the sample.
2.4. Extraction of lipophilic fraction
The lipophilic fraction of lentils was extracted from the finely
ground samples following a slightly modified version of the method
described by Villalobos Solis, Patel, Orsat, Singh, and Lefsrud (2013).
Briefly, ca. 1 g lentil sample was accurately weighed and placed in a
15-mL Teflon-lined screw-capped glass centrifuge tube and
extracted with 10 mL hexane/isopropanol (3:2, v/v) at room temper-
ature with constant rolling on a rotary shaker (Scientific Industries
Inc., Bohemia, NY) at 150 rpm for 5 h. The mixture was then vortexed
for 30 s followed by centrifugation (Eppendorf centrifuge 5810R,
Brinkman Instruments Inc., Westbury, NY) at 3000g for 10 min and
the supernatant was collected in a glass centrifuge tube. The residue
was re-extracted with the same method until it was colourless. The
combined supernatant was then evaporated under nitrogen stream
at room temperature, and re-dissolved in 1 mL tetrahydrofuran and
stored at –20 °C before being used for different analyses. All extrac-
tion experiments were conducted under subdued light, and the
extraction tubes were wrapped with aluminium foil to avoid sample
degradation by photooxidation.
2.5. Fatty acid analysis by gas chromatography
The fatty acid composition of the lipophilic fraction extracted
from lentils was analysed according to the method described by
Kramer et al. (1997). Fatty acid methyl esters (FAME) were pre-
pared using base-catalysed reactions. Briefly, 10 mg of lipids were
placed in a 15-mL glass tube equipped with Teflon-lined screw cap
and dissolved in 80 lL of toluene. One millilitre of NaOCH3/meth-
anol (0.5 N) was then added for the methylation and heated for
30 min at 50 °C. After cooling to room temperature, 1 mL of water
was added to the solution, and the esters were extracted with 2 mL
of hexane. FAME were analysed by GC (Model 6890; Hewlett–
Packard, Palo Alto, CA) using a CP-Sil 88 WCOT fused silica column
(100 m  0.25 mm i.d.  0.2 lm film thickness; Chrompack, Mid-
dleburg, The Netherlands). Column was operated at 45 °C for
4 min, then temperature-programmed at 13 °C/min to 175 °C, held
for 27 min, programmed at 4 °C/min to 215 °C, and finally held for
31 min; total run time was 80 min, the sample injection volume
was 1 ll, injection mode was splitless. Flame ionisation detection
(FID) was used.
2.6. Determination of tocopherols by HPLC
Tocopherols were analysed following a published procedure
with slight modification (Li, Tsao, Yang, Kramer, & Hernandez,
2007). Agilent Technologies 1100 series HPLC system equipped
with a quaternary pump, a degasser, a thermostatic autosampler,
a diode-array detector (DAD) and a fluorescence detector was used
for the analysis of tocopherols in the lipophilic extracts of lentils.
The latter detector was used for the quantification of individual
tocopherols. The detection was set at 295 nm for DAD, and
kexc = 290 nm, kemis = 330 nm, for the fluorescence detector. A Phe-
nomenex (Torrance, CA) silica column (250  4.6 mm, 5 lm) was
used for separation. The mobile phase contained 10% tert-butyl
methyl ether in hexane (v/v), and the flow rate was kept constant
298 B. Zhang et al. / Food Chemistry 161 (2014) 296–304
at 1.0 mL/min for a total run time of 25 min. The sample injection
volume was 10 lL.
2.7. Total carotenoid content
The total carotenoid contents (TCC) in the lentils were deter-
mined according to the method of Ndolo et al. with some modifi-
cation (Ndolo & Beta, 2013). The absorbance was measured at
450 nm using a UV–Vis plate reader (EL 340; Bio-Tek Instruments
Inc., Winooski, VT). The TCC was calculated on the basis of authen-
tic lutein, which is the main carotenoid in lentil, using the follow-
ing equation and expressed as lg/g dry weight (DW) sample.
C ¼ ðV  AÞ=ðS  WÞ½lg=g
where A is the absorbance reading at 450 nm, S is the regression
coefficient (the number that expresses the relationship which is
created based on concentration of lutein standard dilutions in lg/
mL and the absorbance), V is the total volume of extract (mL), W
is the sample dry weight (g).
2.8. Carotenoid composition by HPLC
The carotenoids in extracts were separated and quantified by
the same Agilent Technologies 1100 series HPLC system as stated
above. Identification of the compounds followed the method as
described by Abdel-Aal et al., with some modification (Abdel-Aal
et al., 2007). Separation was done using a C30 YMC Carotenoid col-
umn (250  4.6 mm, 5 lm; Waters, Mississauga, ON). The column
temperature was kept at 35 °C and eluted with a gradient mobile
system consisting of (A) methanol/methyl tert-butyl ether/distilled
water (81:15:4, v/v/v) and (B) methyl tert-butyl ether/methanol
(90:10, v/v). The linear gradient program was set as follows:
100% to 0% A within 60 min and then back to the original solvent
composition (100% A) within 5 min. The flow rate was 1 mL/min
and the UV/Vis absorbance of the peaks was detected and mea-
sured at 450 nm. The carotenoids were identified based on match-
ing retention time, similarity in elution order and UV/Vis spectra
with those of standards and spectra reported in the literature. All
carotenoids were quantified based on the corresponding curves
of the all-trans standards.
2.9. Antioxidant assays
2.9.1. DPPH assay
The DPPH radical-scavenging activity of the lipid extracts was
determined spectrophotometrically on a UV–Vis plate reader
according to the method described by Li et al. with slightly modi-
fications (Li, Deng, Liu, Loewen, & Tsao, 2013). Briefly, aliquots of
280 lL of 65 lM DPPH in methanol were mixed with 20 lL of
the standard Trolox or lipid extracts of lentils diluted 5-fold with
methanol in a 96-well plate. The mixtures were reacted for 3 h
under subdued light at room temperature. The absorbance of DPPH
radicals was read at 540 nm. All samples were tested in triplicate.
The scavenging of DPPH radical was calculated as equivalent units
of Trolox (lmol TE/g DW).
2.9.2. Photochemiluminescence (PCL) assay
The superoxide radical-scavenging capability of lipid extracts of
lentils was assayed using the ACL protocol with the PHOTOCHEMÒ
system (Analytik Jena, Berlin, Germany). The principle of this
method is based on the inhibition of photo-induced autoxidation
of luminol by antioxidants mediated from the radical superoxide
anion. The PCL assay was conducted via the method previously
described (Li, Deng, Liu, Loewen, & Tsao, 2012). Complete reagent
kits were purchased from the manufacturer. Briefly, the assay
was a mixture of 2.3 mL of reagent 1 (sample solvent), 0.2 mL of
reagent 2 (reaction buffer), and 25 lL of diluted reagent 3 (lumi-
nol) and reagent 4 (Trolox) for the calibration curve. To measure
the antioxidant activity of a sample, reagent 4 was simply replaced
by a sample solution. Trolox was used as a reference to evaluate
the superoxide radical-scavenging capability and the antioxidant
activities were expressed as lmol Trolox equivalent/g dry weight
lentil (lmol TE/g DW).
2.10. Statistical analysis
The analytical data were expressed as mean ± SD of triplicate
analysis for TCC and HPLC assay of independent extractions. One-
way analysis of variance (ANOVA) was used to compare the means.
Differences were considered significant at p < 0.05. All statistical
analyses were performed with Statistix for Windows version 9.0
(Analytical Software, Tallahassee, FL).
3. Results and discussion
3.1. Colorimetric study
Colour is one of the most important aspects in making food
choices, and a key characteristic for studying pigments in food.
As shown in Table 1, the lightness value L⁄ varied from 74.37 to
85.15 for the 20 cultivars, but was generally similar between the
red and green lentils. The index a⁄ varied significantly between
the two groups of lentils. As expected, all red lentils had a positive
a⁄ value whereas all green lentils had negative values. The hue
angle H measures the absence of red colour in lentils. A hue of
180° represents pure green and a hue of 0° represents pure red
(Arias, Lee, Logendra, & Janes, 2000). It was not surprising that
the hue angle H was greater in green lentils (ranging from 90.10
to 96.70) than that of red lentils (ranging from 71.23 to 80.37). In
general, there was a good correlation between colour parameters
and the carotenoid content. Previous studies have corroborated
that a⁄/b⁄ ratio was positively related with the lycopene content
(Georgé et al., 2011). Similar correlation was found between b⁄
and total lutein content in red lentils of the present study
(y = 2.2872x – 24.652, r2 = 0.8749). For example, Rouleau which
had the lowest total lutein content (Table 4) possessed the lowest
b⁄ value (12.57), while Impact, which had the highest total lutein
content (Table 4), was a close second to the highest b⁄ value
(16.90) in the red group (Table 1). The correlation was even better
in green lentils (y = 1.0799x – 7.2354, r2 = 0.8859). These results
indicate that the colour index b⁄ could be used to predict the lutein
content in lentils. Colour is a very important marketing factor
affecting the buying decision of consumers, and a key quality
parameter of food processing and product development. Compared
to chemical methods, colorimetric measurement is more conve-
nient for quick estimation of carotenoids content.
3.2. Fatty acids composition
Lipids and oils are important storage forms of carbon in many
seeds. The oil contents and composition of fatty acids of lentils
are summarised in Table 2. The oil yield determined for the 20 len-
til seed powders varied from 1.52% to 2.95%, which was greater
than those found in the literature (Boschin & Arnoldi, 2011).
The difference could be attributed to the type of cultivar, geograph-
ical origin and the genotype of the tested lentils. Crops grown
in a northern climate tend to have higher oil content (the
so-called northern vigour phenomenon) (Sediqi, 2012). The
lipophilic extracts mainly contained unsaturated fatty acids
(UFA) (77.46–81.72%) including mono-unsaturated fatty acids
(MUFA) (21.72–29.10%) and polyunsaturated fatty acids (PUFA)
 B. Zhang et al. / Food Chemistry 161 (2014) 296–304 299

Table 1
Colorimetric parameters of lentils.A

Cultivar L⁄ a⁄ b⁄ c H
Red lentils
Blaze 74.37 ± 1.27 3.88 ± 0.38 13.14 ± 1.78 14.51 ± 0.65 74.13 ± 0.40
Redcliff 78.54 ± 0.48 2.88 ± 0.20 16.84 ± 1.06 17.08 ± 1.08 80.37 ± 0.31
Maxim 79.62 ± 0.61 3.05 ± 0.13 14.69 ± 0.43 15.00 ± 0.45 78.37 ± 0.15
Rouleau 77.52 ± 0.40 4.97 ± 0.41 12.57 ± 1.22 13.51 ± 1.28 71.23 ± 0.06
Redbow 79.03 ± 0.35 3.61 ± 0.06 15.74 ± 0.16 16.15 ± 0.17 77.13 ± 0.06
Redberry 80.65 ± 0.70 3.87 ± 0.07 13.55 ± 0.26 14.08 ± 0.27 74.13 ± 0.25
Impact 77.01 ± 0.51 4.07 ± 0.08 16.90 ± 0.44 17.38 ± 0.45 76.53 ± 0.06
Imperial 80.17 ± 1.68 4.03 ± 0.09 18.33 ± 0.13 18.76 ± 0.12 77.63 ± 0.35
Rosetown 80.24 ± 0.37 3.74 ± 0.25 14.75 ± 0.64 15.22 ± 0.68 75.87 ± 0.32
Dazil 80.47 ± 0.55 3.23 ± 0.13 15.74 ± 0.22 16.06 ± 0.25 78.47 ± 0.25
Green lentils
Imvincible 80.85 ± 0.10 –1.51 ± 0.04 18.97 ± 0.68 19.03 ± 0.68 94.47 ± 0.29
Greenland 82.31 ± 0.73 –1.07 ± 0.06 16.27 ± 0.47 16.30 ± 0.47 93.70 ± 0.17
Asterix 81.54 ± 0.09 –1.29 ± 0.08 15.80 ± 0.38 15.85 ± 0.38 94.60 ± 0.40
Imigreen 83.56 ± 1.12 –2.15 ± 0.03 18.09 ± 0.40 18.22 ± 0.40 96.70 ± 0.17
Impower 85.15 ± 0.37 –2.01 ± 0.07 18.15 ± 0.64 18.25 ± 0.64 96.27 ± 0.25
Improve 80.58 ± 0.16 –0.29 ± 0.06 12.91 ± 0.34 12.91 ± 0.34 91.20 ± 0.26
Sovereign 78.82 ± 0.54 –0.22 ± 0.03 10.93 ± 0.11 10.93 ± 0.11 90.10 ± 0.17
Milestone 77.32 ± 1.25 –0.69 ± 0.02 15.46 ± 0.98 15.47 ± 0.98 92.47 ± 0.15
Eston 77.56 ± 0.76 –0.18 ± 0.02 13.56 ± 0.24 13.56 ± 0.24 90.67 ± 0.06
Plato 83.07 ± 0.79 –0.11 ± 0.08 13.56 ± 0.59 13.66 ± 0.59 90.43 ± 0.31
A
Data are expressed as the mean ± SD, n = 3.

(50.62–58.08%), with an insignificant amount of saturated fatty
acids (SFA). In all lentil cultivars, 18:2n–6 was the dominant fatty
acid ranging from 40.73% to 47.06%, followed by 18:1 (20.11–
28.00%), 16:0 (12.67–14.82%) and 18:3n–3 (9.00–13.28%) (Table 2).
These results agree with those reported by Zia-Ul-Haq et al. in their
study on 4 breeding lines grown in Pakistan (Zia-Ul-Haq et al.,
2011). The high levels of unsaturated fatty acids make lentil a suit-
able legume for nutritional applications. The potential benefits of
PUFA include prevention of cardiovascular diseases and other
health problems (Ajayi & Ajayi, 2009; Faremi & Ekanem, 2011).
and pulse, such as oats, barley, quinoa, and common beans
(Peterson, 1995; Ryan, Galvin, O’Connor, Maguire, & OBrien,
2007). When lentils are incorporated into diets and consumed reg-
ularly, they can therefore contribute a significant portion to the
recommended intake of vitamin E (i.e. 15 mg/day for adults in
USA). The minimum daily intake of vitamin E is set at 3 and
4 mg for women and men, respectively (Monsen, 2000).
3.4. Identification and quantification of carotenoids in lentils

The carotenoid composition of lentils has not been well studied.

3.3. Separation, identification and quantification of tocopherols in
lentils by HPLC
Lentils are also considered to be an excellent source of tocophe-
rols. Tocopherols (vitamin E) are essential antioxidants that have
been known to possess positive effects on human health. Among
the major tocopherols, a-, c- and d-tocopherols have been com-
monly reported in lentils (Grela & Günter, 1995), b-tocopherol
has only been identified in a very limited number of lentil cultivars
(Fernandez-Orozco, Zieliński, & Piskuła, 2003). a-Tocopherol is
believed to be the main contributor to the vitamin E activity
(Boschin & Arnoldi, 2011), whereas c-tocopherol is considered
most efficient in preventing food autoxidation, with a higher anti-
oxidant capacity than a-tocopherol in some model systems
(Amarowicz & Pegg, 2008; Boschin & Arnoldi, 2011). Moreover,
recent studies have shown that c-tocopherol has some potential
functions in the detoxification of nitrogen dioxide and protection
against cardiovascular disease (Amarowicz & Pegg, 2008).
Table 3 shows the concentration of tocopherols in all lentil vari-
eties. c-Tocopherol was the most abundant isomer, accounting for
96–98% of the total tocopherol content, followed by a- and d-
tocopherol. The total and individual tocopherols differed signifi-
cantly among all lentil cultivars. Imvincible and Greenland con-
tained the highest total tocopherols at 63.6 and 64.4 lg/g dry
weight (DW) of seeds, respectively, whereas the lowest total
tocopherol content was found in Sovereign (37.6 lg/g DW) and
Plato (37.4 lg/g DW). These values are consistent with those
reported by Boschin and Arnoldi (2011). Total tocopherol content
of lentils is significantly higher than that reported for other cereals
In this study, we compared four typical solvent systems used in the
literature for lipid or carotenoid extraction from legumes or grains,
and optimised one solvent system for lentil samples. A solvent sys-
tem containing hexane/isopropanol (3:2, v/v) had the highest
carotenoids contents as compared with other three solvents (hex-
ane with water-saturated butanol, MTBE, MTBE with tetrahydrofu-
ran). The identification of individual carotenoids was accomplished
by comparing the retention time, elution order and UV/Vis spectral
data with those of authentic standards and those reported in the
literature or by this laboratory (Abdel-Aal et al., 2007; Li et al.,
2013). The HPLC chromatograms of mixed carotenoid standards
(lutein and zeaxanthin) and a typical lentil lipophilic extract are
shown in Fig. 1. The two major peaks in the extract were identified
as lutein and zeaxanthin. No b-carotene was detected in any of the
20 lentil cultivars. The total carotenoid contents (TCC) and individ-
ual carotenoid compositions of the 20 lentil cultivars collected in
Canada are shown in Table 4. The TCC as determined by the color-
imetric method ranged from 5.32 to 28.1 lg/g DW, while the total
carotenoid index (TCI, sum of all carotenoid concentrations by
HPLC) ranged from 4.64 to 19.6 lg/g DW. A strong correlation
(r2 = 0.9101, p < 0.05) and relatively close results were observed
between the TCC and TCI, suggesting lentils are good source of
carotenoids, particularly lutein and zeaxanthin. The highest TCC
was found in the red lentil cultivar Impact (28.1 lg/g DW), fol-
lowed by red lentil cultivar Imperial (21.9 lg/g DW) and green len-
til cultivar Imvincible (19.0 lg/g DW), while the lowest TCC was
observed in red lentil cultivar Rouleau (5.32 lg/g DW) and green
lentil cultivar Sovereign (6.02 lg/g DW). Although the average
TCC value (15.1 lg/g DW) for red lentils was higher than that for
300 B. Zhang et al. / Food Chemistry 161 (2014) 296–304

Table 2
Fatty acid composition (relative percent) of the lipophilic extracts from different lentil cultivars.

Fatty acid Cultivar (red)

Blaze Redcliff Maxim Rouleau Redbow Redberry Impact Imperial Rosetown Dazil
14:0 0.14 ± 0.01 0.25 ± 0.01 0.22 ± 0.01 0.23 ± 0.01 0.33 ± 0.02 0.20 ± 0.01 0.23 ± 0.01 0.26 ± 0.02 0.22 ± 0.01 0.23 ± 0.01
15:0 0.17 ± 0.01 0.13 ± 0.01 0.14 ± 0.01 0.15 ± 0.02 0.14 ± 0.01 0.14 ± 0.01 0.16 ± 0.02 0.15 ± 0.02 0.14 ± 0.01 0.17 ± 0.02
16:0 14.34 ± 0.15 14.82 ± 0.20 13.60 ± 0.14 13.19 ± 0.17 14.16 ± 0.19 13.32 ± 0.12 14.62 ± 0.15 13.64 ± 0.17 13.89 ± 0.15 14.35 ± 0.21
16:1 0.62 ± 0.04 0.08 ± 0.00 0.10 ± 0.01 0.23 ± 0.02 0.32 ± 0.02 0.09 ± 0.00 0.13 ± 0.01 0.11 ± 0.01 0.09 ± 0.01 0.64 ± 0.05
17:0 0.15 ± 0.01 0.12 ± 0.01 0.24 ± 0.01 0.13 ± 0.01 0.14 ± 0.01 0.14 ± 0.01 0.24 ± 0.02 0.22 ± 0.02 0.13 ± 0.01 0.21 ± 0.01
17:1 0.60 ± 0.02 0.08 ± 0.01 0.16 ± 0.01 0.25 ± 0.01 0.28 ± 0.02 0.08 ± 0.01 0.19 ± 0.01 0.14 ± 0.01 0.07 ± 0.00 0.59 ± 0.03
18:0 1.73 ± 0.06 1.29 ± 0.04 1.66 ± 0.06 1.15 ± 0.07 1.51 ± 0.05 1.31 ± 0.07 1.50 ± 0.04 1.73 ± 0.08 1.13 ± 0.03 1.24 ± 0.05
18:1 20.11 ± 0.15 21.90 ± 0.31 23.86 ± 0.19 27.25 ± 0.52 25.38 ± 0.36 24.15 ± 0.28 21.15 ± 0.23 25.45 ± 0.53 22.13 ± 0.35 22.06 ± 0.19
19:0 0.09 ± 0.01 0.03 ± 0.00 0.03 ± 0.00 0.02 ± 0.00 0.02 ± 0.00 0.03 ± 0.00 0.03 ± 0.00 0.03 ± 0.00 0.02 ± 0.00 0.02 ± 0.00
18:2n-6 44.44 ± 1.35 44.13 ± 1.51 43.39 ± 1.33 41.67 ± 1.45 40.73 ± 1.09 45.41 ± 1.67 42.86 ± 1.28 42.17 ± 0.97 47.06 ± 1.68 41.56 ± 1.06
20:0 0.86 ± 0.03 0.53 ± 0.03 0.45 ± 0.02 0.48 ± 0.01 0.65 ± 0.03 0.42 ± 0.02 0.57 ± 0.03 0.57 ± 0.02 0.43 ± 0.02 0.70 ± 0.01
20:1 0.20 ± 0.01 0.91 ± 0.05 0.74 ± 0.02 0.91 ± 0.03 0.89 ± 0.03 0.79 ± 0.02 0.71 ± 0.02 0.77 ± 0.01 0.91 ± 0.03 0.81 ± 0.03
18:3n-3 13.28 ± 0.17 11.66 ± 0.12 11.69 ± 0.16 9.32 ± 0.11 10.44 ± 0.09 9.84 ± 0.11 12.68 ± 0.11 10.58 ± 0.14 9.67 ± 0.10 12.11 ± 0.11
21:0 0.56 ± 0.02 0.21 ± 0.01 0.17 ± 0.01 0.24 ± 0.02 0.26 ± 0.01 0.21 ± 0.01 0.22 ± 0.02 0.20 ± 0.01 0.20 ± 0.01 0.55 ± 0.01
20:2n-6 0.11 ± 0.01 0.08 ± 0.00 0.08 ± 0.00 0.08 ± 0.01 0.08 ± 0.00 0.08 ± 0.00 0.09 ± 0.00 0.08 ± 0.00 0.09 ± 0.00 0.10 ± 0.01
22:0 0.51 ± 0.03 0.57 ± 0.02 0.42 ± 0.02 0.47 ± 0.01 0.49 ± 0.02 0.47 ± 0.03 0.49 ± 0.02 0.41 ± 0.02 0.49 ± 0.02 0.40 ± 0.01
20:3n-6 0.11 ± 0.01 0.12 ± 0.01 0.15 ± 0.02 0.12 ± 0.01 0.13 ± 0.00 0.16 ± 0.01 0.18 ± 0.01 0.12 ± 0.00 0.13 ± 0.00 0.12 ± 0.00
22:1 0.14 ± 0.01 0.17 ± 0.01 0.13 ± 0.00 0.17 ± 0.01 0.15 ± 0.01 0.13 ± 0.00 0.14 ± 0.00 0.11 ± 0.00 0.18 ± 0.02 0.13 ± 0.01
23:0 0.29 ± 0.01 0.23 ± 0.02 0.19 ± 0.01 0.19 ± 0.01 0.22 ± 0.02 0.20 ± 0.01 0.20 ± 0.01 0.16 ± 0.01 0.20 ± 0.01 0.23 ± 0.02
24:0 0.47 ± 0.02 0.42 ± 0.02 0.38 ± 0.02 0.36 ± 0.01 0.45 ± 0.02 0.35 ± 0.02 0.41 ± 0.01 0.32 ± 0.01 0.33 ± 0.02 0.36 ± 0.01
20:5n-3 0.14 ± 0.01 0.02 ± 0.00 0.02 ± 0.00 0.02 ± 0.00 0.02 ± 0.00 0.04 ± 0.00 0.11 ± 0.01 0.11 ± 0.00 0.02 ± 0.00 0.12 ± 0.00
24:1 0.05 ± 0.00 0.05 ± 0.01 0.04 ± 0.00 0.06 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.04 ± 0.00
26:0 0.50 ± 0.02 0.41 ± 0.01 0.37 ± 0.02 0.59 ± 0.02 0.50 ± 0.01 0.55 ± 0.02 0.50 ± 0.02 0.58 ± 0.01 0.49 ± 0.01 0.35 ± 0.01
Others 0.39 ± 0.01 1.79 ± 0.07 1.77 ± 0.04 2.72 ± 0.10 2.67 ± 0.05 1.85 ± 0.07 2.55 ± 0.05 2.06 ± 0.06 1.94 ± 0.04 2.91 ± 0.08
Oil yield (%) 2.46 ± 0.19 1.89 ± 0.15 1.52 ± 0.17 2.18 ± 0.16 2.95 ± 0.15 1.96 ± 0.15 2.13 ± 0.17 2.09 ± 0.15 2.05 ± 0.18 2.60 ± 0.14
R SFA 19.81 ± 0.38 19.01 ± 0.38 17.87 ± 0.33 17.20 ± 0.36 18.87 ± 0.39 17.34 ± 0.33 19.17 ± 0.35 18.27 ± 0.39 17.67 ± 0.29 18.81 ± 0.37
R MUFA 21.72 ± 0.23 23.19 ± 0.39 25.03 ± 0.23 28.87 ± 0.59 27.06 ± 0.44 25.28 ± 0.31 22.36 ± 0.27 26.61 ± 0.56 23.42 ± 0.41 24.27 ± 0.31
R PUFA 58.08 ± 1.55 56.01 ± 1.64 55.33 ± 1.51 51.21 ± 1.58 51.40 ± 1.18 55.53 ± 1.79 55.92 ± 1.41 53.06 ± 1.11 56.97 ± 1.78 54.01 ± 1.18
R UFA 79.80 ± 1.78 79.20 ± 2.03 80.36 ± 1.74 80.08 ± 2.07 78.46 ± 1.62 80.81 ± 2.10 78.28 ± 1.68 79.67 ± 1.67 80.39 ± 2.19 78.28 ± 1.49
14:0 0.21 ± 0.01 0.30 ± 0.02 0.26 ± 0.01 0.24 ± 0.01 0.23 ± 0.01 0.27 ± 0.01 0.26 ± 0.01 0.35 ± 0.03 0.31 ± 0.01 0.28 ± 0.02
15:0 0.17 ± 0.01 0.14 ± 0.01 0.14 ± 0.01 0.17 ± 0.01 0.15 ± 0.01 0.15 ± 0.01 0.14 ± 0.01 0.11 ± 0.01 0.15 ± 0.02 0.13 ± 0.01
16:0 14.76 ± 0.20 13.95 ± 0.18 13.28 ± 0.18 13.69 ± 0.16 13.00 ± 0.12 12.67 ± 0.13 13.70 ± 0.17 14.54 ± 0.17 14.15 ± 0.21 14.18 ± 0.15
16:1 0.66 ± 0.03 0.91 ± 0.03 0.11 ± 0.01 0.10 ± 0.01 0.83 ± 0.05 0.67 ± 0.04 0.09 ± 0.01 0.75 ± 0.08 0.56 ± 0.03 0.09 ± 0.01
17:0 0.21 ± 0.01 0.10 ± 0.01 0.12 ± 0.01 0.22 ± 0.01 0.23 ± 0.01 0.22 ± 0.02 0.10 ± 0.01 0.13 ± 0.01 0.13 ± 0.01 0.12 ± 0.01
17:1 0.62 ± 0.04 0.83 ± 0.03 0.09 ± 0.00 0.14 ± 0.01 0.85 ± 0.02 0.72 ± 0.04 0.07 ± 0.01 0.69 ± 0.04 0.48 ± 0.02 0.07 ± 0.01
18:0 1.10 ± 0.05 1.26 ± 0.03 1.15 ± 0.07 1.16 ± 0.05 1.06 ± 0.04 1.10 ± 0.05 1.02 ± 0.04 1.44 ± 0.06 1.38 ± 0.03 1.07 ± 0.02
18:1 21.17 ± 0.47 25.32 ± 0.44 22.92 ± 0.56 26.10 ± 0.68 23.25 ± 0.33 25.41 ± 0.74 28.00 ± 0.82 24.35 ± 0.39 23.99 ± 0.46 25.57 ± 0.63
19:0 0.03 ± 0.00 0.04 ± 0.00 0.02 ± 0.00 0.04 ± 0.01 0.02 ± 0.00 0.05 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.04 ± 0.00
18:2n-6 42.92 ± 1.24 40.97 ± 1.18 45.46 ± 1.39 41.49 ± 1.15 42.12 ± 1.03 42.52 ± 1.31 41.64 ± 1.54 41.49 ± 1.62 42.64 ± 1.37 41.95 ± 1.22
20:0 0.84 ± 0.03 0.80 ± 0.02 0.39 ± 0.02 0.48 ± 0.02 0.88 ± 0.03 0.67 ± 0.01 0.34 ± 0.01 0.95 ± 0.03 0.68 ± 0.02 0.42 ± 0.02
20:1 0.78 ± 0.02 0.85 ± 0.02 0.84 ± 0.02 0.76 ± 0.03 0.76 ± 0.01 0.71 ± 0.01 0.78 ± 0.02 0.80 ± 0.03 0.77 ± 0.03 0.86 ± 0.02
18:3n-3 11.33 ± 0.12 9.39 ± 0.11 11.86 ± 0.09 11.38 ± 0.13 11.28 ± 0.15 9.83 ± 0.09 9.35 ± 0.13 9.00 ± 0.08 9.49 ± 0.10 10.74 ± 0.12
21:0 0.51 ± 0.02 0.40 ± 0.02 0.17 ± 0.01 0.17 ± 0.01 0.61 ± 0.03 0.26 ± 0.01 0.11 ± 0.01 0.39 ± 0.02 0.32 ± 0.01 0.17 ± 0.01
20:2n-6 0.10 ± 0.00 0.08 ± 0.01 0.10 ± 0.01 0.08 ± 0.00 0.08 ± 0.00 0.07 ± 0.00 0.07 ± 0.00 0.08 ± 0.00 0.08 ± 0.00 0.09 ± 0.00
22:0 0.37 ± 0.01 0.40 ± 0.02 0.39 ± 0.02 0.44 ± 0.02 0.37 ± 0.01 0.43 ± 0.01 0.41 ± 0.02 0.58 ± 0.03 0.49 ± 0.02 0.51 ± 0.02
20:3n-6 0.12 ± 0.01 0.15 ± 0.01 0.12 ± 0.01 0.13 ± 0.01 0.14 ± 0.01 0.11 ± 0.00 0.11 ± 0.00 0.11 ± 0.00 0.13 ± 0.01 0.12 ± 0.00
22:1 0.14 ± 0.01 0.13 ± 0.00 0.15 ± 0.01 0.17 ± 0.01 0.12 ± 0.00 0.13 ± 0.00 0.12 ± 0.00 0.11 ± 0.00 0.14 ± 0.01 0.14 ± 0.01
23:0 0.19 ± 0.01 0.18 ± 0.01 0.17 ± 0.01 0.18 ± 0.01 0.17 ± 0.00 0.19 ± 0.01 0.17 ± 0.01 0.20 ± 0.01 0.20 ± 0.01 0.21 ± 0.01
24:0 0.30 ± 0.01 0.37 ± 0.02 0.29 ± 0.01 0.36 ± 0.01 0.34 ± 0.01 0.40 ± 0.01 0.38 ± 0.01 0.44 ± 0.01 0.33 ± 0.01 0.47 ± 0.02
20:5n-3 0.11 ± 0.00 0.03 ± 0.00 0.02 ± 0.00 0.02 ± 0.00 0.18 ± 0.01 0.02 ± 0.00 0.03 ± 0.00 0.05 ± 0.00 0.02 ± 0.00 0.02 ± 0.00
24:1 0.03 ± 0.00 0.05 ± 0.00 0.05 ± 0.01 0.05 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.04 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.04 ± 0.00
26:0 0.55 ± 0.01 0.66 ± 0.03 0.33 ± 0.01 0.56 ± 0.01 0.62 ± 0.02 0.58 ± 0.03 1.04 ± 0.04 0.84 ± 0.02 0.98 ± 0.03 0.63 ± 0.02
Others 2.78 ± 0.05 2.69 ± 0.11 1.57 ± 0.05 1.87 ± 0.04 2.67 ± 0.10 2.78 ± 0.06 2.00 ± 0.05 2.53 ± 0.09 2.50 ± 0.06 2.08 ± 0.08
Oil yield (%) 2.61 ± 0.16 2.63 ± 0.16 1.93 ± 0.17 1.69 ± 0.13 2.41 ± 0.15 2.28 ± 0.15 2.53 ± 0.14 2.42 ± 0.17 1.96 ± 0.17 1.85 ± 0.13
R SFA 19.24 ± 0.37 18.60 ± 0.37 16.71 ± 0.36 17.71 ± 0.33 17.68 ± 0.29 16.99 ± 0.28 17.70 ± 0.34 20.01 ± 0.40 19.16 ± 0.38 18.23 ± 0.31
R MUFA 23.40 ± 0.57 28.09 ± 0.52 24.16 ± 0.62 27.32 ± 0.74 25.85 ± 0.41 27.68 ± 0.83 29.10 ± 0.86 26.73 ± 0.54 25.98 ± 0.55 26.77 ± 0.68
R PUFA 54.58 ± 1.37 50.62 ± 1.31 57.56 ± 1.50 53.10 ± 1.29 51.80 ± 1.20 52.55 ± 1.40 51.20 ± 1.67 50.73 ± 1.70 52.36 ± 1.48 52.92 ± 1.34
R UFA 77.98 ± 1.94 78.71 ± 1.83 81.72 ± 2.12 80.42 ± 2.03 77.65 ± 1.61 80.23 ± 2.23 80.30 ± 2.53 77.46 ± 2.24 78.34 ± 2.03 79.69 ± 2.02

green lentils (13.1 lg/g DW), no statistically significant difference
was observed between the two colour groups. Similar results were
found in TCI. The slightly higher TCC values than TCI values could
be attributed to the minor carotenoids not quantifiable with the
HPLC methods.
Among the carotenoids, only lutein, zeaxanthin and their iso-
mers were detected in all 20 lentil cultivars. All-trans-lutein, which
accounts for 64–78% of total carotenoids in lentils, was the primary
and predominant carotenoid, followed by all-trans-zeaxanthin,
which constitutes 5–13% of the total carotenoids. Apart from all-
trans isomeric forms, about 20% of the total carotenoids were cis-
isomers, mainly 9-cis-lutein, 9́-cis-lutein and 13-cis-lutein, possibly
formed during harvest, storage and extraction in the laboratory
(Table 4). The total lutein and zeaxanthin contents (total of all-
trans and cis-isomers) ranged from 4.32–17.3 lg/g DW and 0.32–
2.73 lg/g DW, respectively, and were significantly different
(p < 0.05) among investigated lentil cultivars. The cultivar Impact
was found to have the greatest amount of total lutein (17.3 lg/g
DW), while the cultivar Imperial had the highest concentration of
total zeaxanthin. The profiles and concentrations found in this
study were similar with those reported for peas (Edelenbos,
Christensen, & Grevsen, 2001; Holasova, Dostalova, Fiedlerova, &
Horacek, 2009) but markedly higher than most of those reported
for high-lutein grains and cereals (Abdel-Aal et al., 2007; Ndolo &
 B. Zhang et al. / Food Chemistry 161 (2014) 296–304 301

Table 3
Tocopherol composition and contents (lg/g DW) of lipophilic extracts from different lentil cultivars.A

Cultivar a-Tocopherol c-Tocopherol (lg/g DW) d-Tocopherol Total tocopherols

Red lentils
Blaze 0.47 ± 0.03 ef 50.3 ± 2.13 cd 1.25 ± 0.12 g 52.0 ± 2.28 cd
Redcliff 0.74 ± 0.05 i 41.6 ± 3.31 ab 0.58 ± 0.02 e 42.9 ± 3.38 ab
Maxim 0.70 ± 0.08 i 46.5 ± 2.58 bc 0.47 ± 0.06 cd 47.7 ± 2.72 bc
Rouleau 0.45 ± 0.02 ef 43.1 ± 1.59 b 0.59 ± 0.11 e 44.1 ± 1.72 b
Redbow 0.60 ± 0.04 h 41.6 ± 2.34 ab 0.47 ± 0.05 cd 42.7 ± 2.43 ab
Redberry 0.35 ± 0.02 d 54.6 ± 2.62 de 0.43 ± 0.04 b 55.4 ± 2.68 de
Impact 0.62 ± 0.02 h 53.1 ± 4.21 de 0.85 ± 0.10 f 54.6 ± 4.33 de
Imperial 0.43 ± 0.03 e 41.3 ± 3.61 ab 0.59 ± 0.03 e 42.3 ± 3.67 ab
Rosetown 0.90 ± 0.07 J 57.8 ± 3.84 e 0.58 ± 0.07 e 59.3 ± 3.98 e
Dazil 0.58 ± 0.02 gh 41.7 ± 1.06 ab 0.54 ± 0.03 de 42.9 ± 1.11 ab
Green lentils
Imvincible 0.76 ± 0.06 i 61.7 ± 3.52 ef 1.13 ± 0.09 g 63.6 ± 3.67 ef
Greenland 0.45 ± 0.05 ef 63.5 ± 1.87 f 0.39 ± 0.02 ab 64.4 ± 1.94 f
Asterix 0.16 ± 0.01 a 41.0 ± 2.06 ab 0.58 ± 0.06 e 41.7 ± 2.13 ab
Imigreen 0.34 ± 0.04 d 51.9 ± 1.58 cd 0.33 ± 0.02 a 52.6 ± 1.64 cd
Impower 0.20 ± 0.04 ab 42.0 ± 1.25 ab 0.49 ± 0.04 c 42.7 ± 1.33 ab
Improve 0.52 ± 0.01 fg 53.9 ± 2.30 de 0.45 ± 0.07 bc 54.9 ± 2.38 de
Sovereign 0.26 ± 0.02 bc 45.2 ± 2.12 bc 0.50 ± 0.06 cd 45.9 ± 2.20 bc
Milestone 0.35 ± 0.08 d 36.8 ± 1.03 a 0.46 ± 0.04 bc 37.6 ± 1.15 a
Eston 0.34 ± 0.08 d 45.4 ± 1.96 bc 0.58 ± 0.06 e 56.3 ± 2.10 bc
Plato 0.31 ± 0.05 cd 36.3 ± 1.14 a 0.75 ± 0.04 f 37.4 ± 1.23 a
A
Values are mean ± SD, n = 3. Values followed by the different letter in the same column are significantly different (p < 0.05).

tocopherols and carotenoids, were examined using the DPPH and

PCL methods. The DPPH assay measures the scavenging ability of
30 antioxidants towards DPPH radical, while the PCL assay measures
the superoxide radical-scavenging capability of antioxidants.
The antioxidant activities varied significantly among the lipo-
philic extracts of the 20 tested lentils. As shown in Fig. 2, the anti-
oxidant activities in the DPPH assay ranged from 3.61 to 4.48 lmol
TE/g DW, and the PCL values were from 2.73 to 6.23 lmol TE/g DW,
20 respectively. Attempts were made to analyse the correlation
between the concentration of the bioactive components and the
antioxidant activities using the Pearson’s correlation coefficient
(r2). Both carotenoids and tocopherols showed weak but significant
correlation with DPPH activity (r2 = 0.4893 and 0.3259, respec-
tively, data not shown). However, an improved correlation with
DPPH value was observed after combining the content of carote-
10 noids and tocopherols (r2 = 0.6687), indicating both carotenoids
and tocopherols contributed to the DPPH scavenging activity and
there is a synergistic effect between carotenoids and tocopherols.
A strong correlation between the carotenoids of cereal and DPPH
activity was found by others (Ndolo & Beta, 2013); however, con-
sidering the present finding of the relatively high levels of total
tocopherols in lentils, these compounds may play a role in the
0
overall radical-scavenging activity.
A good correlation was observed between the carotenoids con-
0 10 20 30 40 tent and PCL activity with r2 value of 0.8153 (data not shown),
whereas the correlation between the tocopherols content and the
min
PCL activity was extremely weak (r2 = 0.0331). The combination of
Fig. 1. LC-UV/vis chromatograms of carotenoids separated from a standard mixture carotenoids and tocopherols was not correlated with PCL activity
(A) and a typical lentil extract. (B) Peaks: UN = unknown; 1 = 15-cis-lutein; 2 = 13- (r2 = 0.4004). These results indicated that carotenoids, mainly lutein,
cis-lutein; 3 = 130 -cis-lutein; 4 = all-trans-lutein; 5 = all-trans-zeaxanthin; 6 = 9-cis-
rather than tocopherols, contributed to the antioxidant activity
lutein; 7 = 90 -cis-lutein; 8 = 9-cis-zeaxanthin.
potential in the PCL assay. This can be seen with the two cultivars
Beta, 2013). Lutein and zeaxanthin are stronger antioxidants than Imigreen and Impower, which showed similar PCL activity and con-
other commonly found carotenoids and play important roles in tained similar total carotenoid content, but significantly higher toc-
maintaining eye health and can protect cells from attacks by car- opherols for the former (p < 0.05, Tables 3 and 4, Fig. 2). The different
cinogens (Wang et al., 2006). antioxidant activities as observed in the DPPH and PCL assays were
also observed by others and considered to be caused by the different
mechanisms (Li et al., 2007; Wang et al., 2006).
3.5. Antioxidant activities and correlation with bioactive components No correlations were found between the contents of UFA, MUFA
and PUFA and antioxidant activities. This suggests that fatty acids
The antioxidant activities of the lipophilic extracts of lentils, in in lentils do not contribute to the antioxidant activity. Previous
which the major antioxidant components were confirmed to be study also showed that none of the UFA (i.e. 18:1, 18:2n–6,
 302
Table 4
Concentrations (lg/g DW) of carotenoids in different cultivars of lentils (n = 3).A

Compounds Blaze Redcliff Maxim Rouleau Redbow Redberry Impact Imperial Rosetown Dazil
Red lentils
15-cis-lutein 0.40 ± 0.06 a 0.39 ± 0.02 a Trace Trace 0.39 ± 0.12 a Trace 0.39 ± 0.05 a 0.40 ± 0.04 a Trace 0.38 ± 0.11 a
13-cis-lutein 0.54 ± 0.04 bc 0.72 ± 0.07 ef 0.53 ± 0.12 bc 0.40 ± 0.03 a 0.67 ± 0.16 de 0.44 ± 0.08 a 0.77 ± 0.21 f 0.74 ± 0.08ef 0.45 ± 0.09 a 0.68 ± 0.09 de
130 -cis-lutein 0.37 ± 0.03 a 0.37 ± 0.04 a Trace Trace 0.37 ± 0.12 a Trace 0.37 ± 0.05 a 0.37 ± 0.04 a Trace Trace
All-trans-lutein 4.90 ± 0.21 b 9.89 ± 0.53 ef 6.55 ± 0.15 d 3.07 ± 0.16 a 9.46 ± 0.42 ef 3.54 ± 0.23 a 14.3 ± 0.82 h 13.4 ± 0.56 h 6.06 ± 0.15 cd 9.25 ± 0.36 e
All-trans-zeaxanthin 0.40 ± 0.02 b 1.33 ± 0.06 g 0.89 ± 0.04 de 0.25 ± 0.04 a 1.05 ± 0.07 ef 0.31 ± 0.05 ab 2.09 ± 0.13 h 2.45 ± 0.25 i 0.65 ± 0.03 c 0.78 ± 0.15 d
9-cis-lutein 0.52 ± 0.02 ab 0.58 ± 0.03 b 0.45 ± 0.04 a 0.40 ± 0.11 a 0.55 ± 0.05 ab 0.39 ± 0.09 a 0.64 ± 0.22 b 0.57 ± 0.03 ab 0.46 ± 0.03 a 0.51 ± 0.06 ab
90 -cis-lutein 0.48 ± 0.03 a 0.50 ± 0.03 a 0.64 ± 0.09 bc 0.45 ± 0.08 a 0.70 ± 0.13 bcd 0.50 ± 0.12 a 0.81 ± 0.06 de 0.89 ± 0.12 e 0.60 ± 0.06 bc 0.52 ± 0.13 ab

B. Zhang et al. / Food Chemistry 161 (2014) 296–304

9-cis-zeaxanthin 0.08 ± 0.01 a 0.16 ± 0.02 ab 0.11 ± 0.01 a 0.07 ± 0.03 a 0.15 ± 0.04 ab 0.08 ± 0.02 a 0.25 ± 0.05 b 0.28 ± 0.07 b 0.12 ± 0.06 a 0.13 ± 0.04 a
Total lutein 7.21 ± 0.39 c 12.5 ± 0.72 de 8.17 ± 0.40 bc 4.32 ± 0.38 a 12.1 ± 1.00 de 4.87 ± 0.52 a 17.3 ± 1.41 f 16.4 ± 0.87 f 7.57 ± 0.33 bc 11.3 ± 0.75 d
Total zeaxanthin 0.48 ± 0.03 ab 1.49 ± 0.08 e 1.00 ± 0.05 cd 0.32 ± 0.07 a 1.20 ± 0.11 de 0.39 ± 0.07 a 2.34 ± 0.18 f 2.73 ± 0.32 f 0.77 ± 0.09 bc 0.91 ± 0.19 cd
Total cis-carotenoids 2.39 ± 0.19 b 2.72 ± 0.21 b 1.73 ± 0.26 a 1.32 ± 0.25 a 2.83 ± 0.62 b 1.41 ± 0.31 a 3.23 ± 0.64 b 3.25 ± 0.38 b 1.63 ± 0.24 a 1.84 ± 0.43 a
Total trans-carotenoids 5.30 ± 0.23 b 11.2 ± 0.59 e 7.44 ± 0.19 c 3.32 ± 0.20 a 10.5 ± 0.49 de 3.85 ± 0.28 a 16.4 ± 0.95 f 15.8 ± 0.81 f 6.71 ± 0.18 bc 10.0 ± 0.51 d
Total carotenoids index (TCI) 7.69 ± 0.42 b 13.9 ± 0.80 fg 9.17 ± 0.45 c 4.64 ± 0.45 a 13.3 ± 1.11 ef 5.26 ± 0.59 a 19.6 ± 1.59 h 19.1 ± 1.19 h 8.34 ± 0.42 bc 11.9 ± 0.94 de
Total carotenoids content (TCC) 9.58 ± 0.75 bc 18.4 ± 2.21 f 11.8 ± 1.37 c 5.32 ± 1.97 a 18.2 ± 1.99 f 9.71 ± 1.31 bc 28.1 ± 2.34 h 21.9 ± 1.40 g 13.6 ± 1.15 d 14.3 ± 1.03 de
Imvincible Greenland Asterix Imigreen Impower Improve Sovereign Milestone Eston Plato
Green lentils
15-cis-lutein 0.38 ± 0.06 a 0.37 ± 0.05 a Trace Trace Trace Trace Trace Trace Trace Trace
13-cis-lutein 0.67 ± 0.03 de 0.55 ± 0.05 bc 0.61 ± 0.05 cd 0.59 ± 0.14 cd 0.57 ± 0.06 bc 0.49 ± 0.08 ab 0.41 ± 0.10 a 0.49 ± 0.09 ab 0.48 ± 0.02 ab 0.44 ± 0.07 ab
130 -cis-lutein 0.37 ± 0.11 a 1.10 ± 0.12 a Trace 0.38 ± 0.05 a 0.37 ± 0.11 a Trace Trace Trace Trace Trace
All-trans-lutein 11.3 ± 0.62 g 9.01 ± 0.26 e 7.17 ± 0.31 d 9.77 ± 0.51 ef 10.4 ± 0.38 fg 5.28 ± 0.17 bc 3.62 ± 0.16 a 6.27 ± 0.22 cd 6.78 ± 0.46 d 5.59 ± 0.71 bc
All-trans-zeaxanthin 1.04 ± 0.08 ef 0.72 ± 0.11 cd 0.69 ± 0.06 c 0.97 ± 0.03 e 1.19 ± 0.06 fg 0.31 ± 0.03 ab 0.31 ± 0.03 ab 0.56 ± 0.08 bc 0.63 ± 0.12 c 0.49 ± 0.06 b
9-cis-lutein 0.63 ± 0.11 b 0.54 ± 0.08 ab 0.46 ± 0.01 a 0.49 ± 0.06 ab 0.49 ± 0.04 ab 0.43 ± 0.05 a 0.40 ± 0.03 a 0.48 ± 0.05 ab 0.47 ± 0.05 ab 0.43 ± 0.12 a
90 -cis-lutein 0.60 ± 0.06 bc 0.61 ± 0.14 bc 0.52 ± 0.06 a 0.56 ± 0.10 ab 0.57 ± 0.05 ab 0.49 ± 0.02 a 0.48 ± 0.11 a 0.53 ± 0.04 ab 0.55 ± 0.09 ab 0.46 ± 0.05 a
9-cis-zeaxanthin 0.12 ± 0.02 a 0.12 ± 0.02 a 0.11 ± 0.01 a 0.12 ± 0.02 a 0.14 ± 0.01 a 0.09 ± 0.03 a 0.08 ± 0.03 a 0.11 ± 0.05 a 0.12 ± 0.02 a 0.11 ± 0.03 a
Total lutein 13.9 ± 0.99 e 12.2 ± 0.70 d 8.76 ± 0.43 c 11.8 ± 0.86 d 12.4 ± 0.64 d 6.69 ± 0.32 b 4.91 ± 0.40 a 7.77 ± 0.40 bc 8.28 ± 0.62 bc 6.92 ± 0.95 b
Total zeaxanthin 1.16 ± 0.10 de 0.84 ± 0.13 c 0.80 ± 0.07 c 1.09 ± 0.05 d 1.33 ± 0.07 e 0.40 ± 0.06 a 0.39 ± 0.06 a 0.67 ± 0.13 b 0.75 ± 0.14 bc 0.60 ± 0.09 b
Total cis-carotenoids 2.77 ± 0.39 b 3.29 ± 0.46 1.70 ± 0.13 a 2.14 ± 0.37 ab 2.14 ± 0.27 ab 1.50 ± 0.18 a 1.37 ± 0.27 a 1.61 ± 0.23 a 1.62 ± 0.18 a 1.44 ± 0.27 a
Total trans-carotenoids 12.3 ± 0.70 e 9.73 ± 0.37 d 7.86 ± 0.37 c 10.74 ± 0.54 de 11.6 ± 0.44 de 5.59 ± 0.20 b 3.93 ± 0.19 a 6.83 ± 0.30 bc 7.41 ± 0.58 c 6.08 ± 0.77 bc
Total carotenoids index (TCI) 15.1 ± 1.09 g 13.0 ± 0.83 ef 9.56 ± 0.50 bc 12.9 ± 0.91 e 13.7 ± 0.71 ef 7.09 ± 0.38 b 5.30 ± 0.46 a 8.44 ± 0.53 bc 9.03 ± 0.76 c 7.52 ± 1.04 b
Total carotenoids content (TCC) 19.0 ± 1.48 f 19.0 ± 1.66 f 12.5 ± 0.12 cd 16.2 ± 0.17 e 16.4 ± 1.50 e 11.8 ± 0.27 cd 6.20 ± 0.44 ab 8.33 ± 0.98 ab 11.8 ± 0.48 cd 9.92 ± 0.20 bc
A
Values are mean ± SD, n = 3. Trace: determined with trace amounts. Values followed by the different letter in the same row are significantly different (p < 0.05).
 B. Zhang et al. / Food Chemistry 161 (2014) 296–304
A B
Sample cultivars
Sample cultivars
Imvincible
DPPH (µmol TE/g DW)
Fig. 2. Antioxidant activities of the lipophilic extracts of 20 different lentil cultivars. (A) DPPH assay, values are expressed as lmol Trolox equivalent/g DW lentil (lmol TE/g
DW); (B) PCL assay, values are expressed as lmol Trolox equivalent/g DW lentil (lmol TE/g DW); Values are means ± SD, n = 3. Values followed by the same letter in the same
assay are not significantly different (p < 0.05). Striped and dotted rectangles represent green and red lentils, respectively.
18:3n–3) in their free form possessed antioxidant activity (Li et al.,
2007).
In conclusion, in addition to the macronutrients, many micro-
nutrients and phytochemicals in lentils may also be important con-
tributors to human health. By examining the lipophilic
components of the most popular lentil cultivars grown in Canada,
we demonstrated that lentils are an excellent source of essential
fatty acids, tocopherols and carotenoids. This is the first compre-
hensive study to report on carotenoid compositions of lentils.
Lutein is the most abundant carotenoid in lentils, and its higher
concentration than in other high-lutein cereal grains suggests con-
sumption of lentils may help maintain eye health and alleviate
other health problems. Carotenoids and tocopherols, very likely
acting synergistically, were the main contributors to the total anti-
oxidant activities in the lipophilic fraction of the 20 lentil cultivars.
Acknowledgements
We thank Pulse Canada for providing the lentil samples. This
project is supported by the A-Base research (RBPI 1004; 2834) of
Agriculture & Agri-Food Canada and an Agri-Innovation Program
Grant with Pulse Canada (AIP#068).
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