Food

Chemistry
Food Chemistry 102 (2007) 123–128
www.elsevier.com/locate/foodchem

Nutritional assessment of cookies supplemented

with defatted wheat germ
Muhammad Umair Arshad *, Faqir Muhammad Anjum, Tahir Zahoor
Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan

Received 20 February 2006; received in revised form 9 March 2006; accepted 27 April 2006

Abstract

Replacement of wheat flour with defatted wheat germ (DFWG) at levels of 0–25% was investigated for its effect on functional and
nutritional properties of cookies. The crude protein content of DFWG was as high as 27.8% with a highly valuable amino acid profile,
rich in essential amino acids, especially lysine (2.32 g/100 g). The physicochemical and sensory evaluation of cookies, revealed that up to
15% substitution of wheat flour with DFWG produced acceptable cookies similar to the control (100% wheat flour) cookies. The protein
quality of the cookies was assessed through weanling albino rats by feeding a diet of cookies for 10 days, which was formulated to supply
10% protein, with a casein diet as a control. The cookies containing 15% DFWG, were best regarding protein bioavailability in rats. The
protein efficiency ratio (PER), net protein utilization (NPU), biological value (BV) and true digestibility (TD) differed significantly
among diets containing cookies with 0–10% DFWG, and casein diet when fed to rats. Diets containing 15% DFWG have values, of these
parameters, similar to the casein diet.
 2006 Elsevier Ltd. All rights reserved.

Keywords: Defatted wheat germ; Cookies; Physicochemical composition; Protein quality

1. Introduction utilized (Shurpalekar & Rao, 1977). The germ provides

three times as much protein, seven times as much fat, fif-
The protein foods available commercially are obtained teen times as much sugars and six times as much mineral
from a range of animal and plant sources and are used as content than does wheat flour (Rao, Kumar, Rao, & Shur-
functional ingredients (Periago & Vidal, 1998). The paleker, 1980).
increased costs and limited supplies of animal proteins, Defatted wheat germ, after extraction of valuable wheat
have necessitated contemporary research efforts geared germ oil, is a high nutritive value protein material, which
towards the study of food properties and potential utiliza- contains about 30% protein (Ge, Sun, Ni, & Cai, 2000).
tion of protein from locally available food crops, especially Wheat germ protein has been classed with effectively supe-
from under-utilized or relatively neglected high protein oil- rior animal proteins and is rich in amino acids, especially
seeds and legumes (Enujiugha & Ayodele-Oni, 2003). the essential amino acids, lysine, methionine, and threo-
Wheat germ protein is reported to have a high nutritive nine, in which many cereals are deficient (Yiqiang, Aidong,
value, comparable to that of animal proteins. In the wheat & Tongyi, 1999). Therefore, wheat germ is a potential
grain, most nutrients, with the exception of starch, are con- nutritious food supplement; in particular, defatted wheat
centrated in germ but most of this is generally used in ani- germ is a source of natural high grade protein, which
mal feed formulations, due to which the precious wheat makes it a good enrichment component for many foods
germ source has not been amply, rationally, and efficiently (Moss, Murray, & Stenvert, 1984).
The enrichment of cereal-based foods with oilseed and
*
Corresponding author. legume protein has received considerable attention. Wheat
E-mail address: [email protected] (M.U. Arshad). bread and cookies are widely accepted and consumed in

0308-8146/$ - see front matter  2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2006.04.040
124 M.U. Arshad et al. / Food Chemistry 102 (2007) 123–128

many developing countries and therefore offer a valuable Cookies were prepared according to the procedure
supplementation vehicle for nutritional improvement; how- described by McWatters et al. (2003) with slight modifica-
ever, cookies have been suggested as a better use of com- tions. The basic ingredients used were 380 g of flour blend,
posite flour than bread because of their ready-to-eat 100 g vegetable shortening, 225 g of granulated cane sugar,
form, wide consumption and relatively long shelf-life 21 g of beaten whole egg, 3.75 g of salt, and 1.8 g of baking
(Lorens, Dilsaver, & Wolt, 1979). Protein enriched cookies powder. The dry ingredients were weighed and mixed thor-
are attractive for target areas, such as child-feeding pro- oughly in a bowl by hand for 3–5 min. Shortening was
grammes, low-income groups and disaster relief operations added and rubbed in until uniform. The egg was added
(Claughton & Pearce, 1989). Cookies with these character- and dough was thoroughly kneaded in a mixer for 5 min.
istics have been produced from blends of wheat and cow- The dough was rolled thinly on a sheeting board to a uni-
pea (McWatters, Ouedraogo, Resurrection, Hung, & form thickness (8.0 mm) and cut out using a round scorn
Philips, 2003) or soybean and wheat (Shrestha & Noom- cutter to a diameter of 35.0 mm. The cut out dough pieces
horm, 2002). However, there is no information on the use were baked on greased pans at 160 C for 15 min in a bak-
of DFWG in cookie-making. In this study, we attempted ing oven. The prepared cookies were cooled to room tem-
to assess the suitability of DFWG utilization for improve- perature (30 ± 2 C) and packed in high density
ment in quality and nutritive value of cookies, and to polyethylene bags.
strengthen the utilization of wheat germ.
2.4. Chemical composition of flours and cookies
2. Materials and methods
Amino acid content of DFWG was determined by using
2.1. Raw materials and treatment an autoanalytic apparatus according to the method of Yu
(1994). All reagents used were of analytical grade (BDH
Wheat was purchased from the Ayub Agricultural Chemicals, Poole, UK).
Research Institute (AARI), Faisalabad. Straight grade The moisture, crude protein, crude fat, total ash and
flour was prepared by a UDY cyclone mill in the Institute crude fibre contents of flours and cookies were determined
of Food Science and Technology, University of Agricul- by AACC (2000). Nitrogen-free extract (NFE) was calcu-
ture, Faisalabad. After milling of wheat, flour samples were lated by difference. The factors, n = 5.70 (for wheat flour),
packed in polypropylene bags and stored at room temper- and n = 6.25 (for cookies) were used for conversion of
ature for further study. Raw wheat germ was procured nitrogen to crude protein. Minerals, including, calcium,
from Sunny Flour Mills, Lahore, Pakistan. Wheat germ iron and potassium, were determined using an atomic
contains several enzymes, such as dipeptidase, proteinase, absorption spectrophotometer, AAS (Model 372, Perkin–
lipase, lipoxidase and phytase. In order to stabilize it, Elmer Ltd., Beaconsfield, UK) by dry-ashing, according
wheat germ was treated by ‘Characteristic Far Red’ ray to the procedure of the AACC (2000). All reagents used
technology to kill its enzymes, during which the tempera- were of analytical grade (BDH Chemicals, Poole, UK).
ture was 130–160 C for 20–25 min. Treatment of wheat
germ, under these conditions, is reported to be sufficient 2.5. Physical and organoleptic evaluation of cookies
enough to deactivate enzyme activity (Zwingelberg & Fret-
zdorff, 1996). Physical parameters, including, diameter, height, weight
and hardness, of cookies were measured on three replicates
2.2. Preparation of DFWG flour and mean values recorded. Cookie diameters and heights
were measured with a vernier caliper. Weights were deter-
Wheat germ oil was extracted by a solvent extraction mined using a Mettler digital top loading balance (PC
technique, using n-hexane (BP. 68 C). DFWG remaining 400; Mettler, Buchi Switzerland).
after oil extraction was crushed and passed through a Cookies were evaluated for colour, flavour, texture and
200-mesh sieve to obtain DFWG flour, which was used overall acceptability, according to the preference method of
as our experimental material. Ihekoronye and Ngoddy (1985). Ten (trained) judges par-
ticipated in the sensory evaluation of the cookies on a 5-
2.3. Blends formulation and preparation of cookies point hedonic scale. The coded cookie samples were ran-
domized and presented to the judges in the midmorning
Blends of wheat flour and DFWG flours containing 0%, on white plates in the sensory evaluation laboratory, Insti-
5%, 10%, 15%, 20% and 25% DFWG flour, on a replace- tute of Food Science and Technology, University of Agri-
ment basis, were prepared. The choice of these levels was culture, Faisalabad.
based on the report of Dreuiter (1978) that the maximum
level of wheat flour substitution that would produce an 2.6. Biological evaluation of protein quality
acceptable baked product was 25%. They were then packed
in polyethylene bags, sealed and stored in a freezer (0–5 C) Based on the results of physicochemical and sensory
until required. evaluation of cookies, four best treatments, including con-
 M.U. Arshad et al. / Food Chemistry 102 (2007) 123–128 125

trol (0% DFWG), were selected for protein quality evalu- 3. Results and discussion
ation by a rat bioassay. Protein diets were formulated
using flours from cookie samples containing 0% (diet A, 3.1. Amino acid profile of DFWG
control), 5% (diet B), 10% (diet C), and 15% (diet D).
Diets were prepared according to AOAC (1995) formula- The amino acid profile of DFWG is shown in Table 2.
tion. Their compositions are shown in Table 1. Weanling The amino acid varieties of DWGP were inclusive, and
male albino rats, 28 days old and weighing between 42 especially rich in essential amino acids and limited amino
and 45 g, were grouped by randomized block design into acids for the human body. Furthermore, the composition
six groups on the basis of weight, such that mean initial ratio of essential amino acids of DFWG, comparable to
weights did not differ by more than ±0.5 g. Each group egg and milk proteins, was basically close to the model
consisted of 10 rats and they were housed in individual value issued by the FAO/WHO with good amino acid equi-
wire-bottom cages that allowed for easy faecal collection librium, which was very significant from a nutritional point
and the measurement of food intake. The temperature of view.
of the animal room was 27 ± 1 C with alternate 12 h.
periods of light and dark. One group of 10 rats was fed 3.2. Chemical analyses of flours and cookies
casein (diet E). The other was fed a protein-free diet
(F), which consisted entirely of the basal diet, while the The chemical compositions of wheat flour and DFWG
remaining four groups were fed the experimental diets flour used for cookies preparation are shown in Table 3.
(A–D). The diets were supplemented with vitamins and Crude protein (28.9%), crude fibre (5.35%), ash (4.52%)
minerals to target requirements (Grant, Dorward, & Pusz- and mineral content are higher in the case of DFWG flour
tai, 1993). Rats were given free access to diet and water. than of wheat flour. A highly significant (P 6 0.05) differ-
Daily records, of weight gain or loss, food and protein ence was also obtained between control and DFWG-
intakes and urinary and faecal output by the rats, were enriched cookies for these chemical parameters (Table 4).
taken. Urinary and fecal nitrogen samples from the Results of the chemical analysis of cookies revealed that
appropriate test diet group were collected and analyzed protein contents of the cookies, prepared from defatted
by the Kjeldahl method (AOAC, 1995). wheat germ flour blends, were significantly higher than
The data obtained from this experiment were used to the protein content of control cookies. The protein content
calculate protein efficiency ratio (PER), true digestibility of the cookies prepared from these blends was also higher
(TD), biological value (BV) and net protein utilization (12.7–16.2%) than those (6–12%) reported for conventional
(NPU) by following the procedures outlined by Pellet cookies (Shrestha & Noomhorm, 2002).
and Young (1980). True digestibility is the apparent digest-
ibility corrected for metabolic N in the fecal material, and
biological value is the percentage of absorbed N that is
retained by the rats (after correction for urinary N and Table 2
Amino acid content of DFWG (g/100 g)
endogenous urinary N). Net protein utilization is derived
from true protein digestibility · biological value per 100. Amino acid Content Amino acid Content
Tryptophan 0.201 Valine 1.40
2.7. Statistical analysis Threonine 0.928 Arginine 4.76
Isoleucine 1.73 Histidine 0.838
Leucine 1.11 Alanine 1.66
Data were statistically analyzed using analysis of vari- Lysine 2.32 Aspartic acid 1.63
ance (ANOVA) according to Steel, Torrie, and Dickey Methionine 0.248 Glutamic acid 5.09
(1997). Means were separated by least significant difference Phenylalanine 1.04 Glycine 1.47
(LSD). Significance was accepted at p 6 0.05. Tyrosine 0.779 Serine 1.43
Total 26.6

Table 1
Composition of experimental diets fed to rats
Table 3
Dietsa Ingredients of diets (g/100 g) Chemical compositions of flours used for cookies preparation
Cookies Corn Mineral Vitamin Corn Casein Components Wheat flour DFWG flour
oil mixture mixture starch
Moisture (%) 10.0 ± 0.4 13.2 ± 0.3
A 83.7 10 2.5 1 2.8 – Crude protein (%) 11.5 ± 0.5 27.8 ± 0.4
B 78.2 10 2.5 1 8.3 – Crude fat (%) 0.92 ± 0.1 0.05 ± 0.01
C 73.7 10 2.5 1 12.8 – Crude fibre (%) 0.90 ± 0.15 5.35 ± 0.3
D 69.1 10 2.5 1 17.4 – Ash (%) 0.74 ± 0.1 4.52 ± 0.2
E – 10 2.5 1 76.5 10 NFE (%) 76.7 ± 1.2 58.0 ± 1.0
F – 10 2.5 1 86.5 – Calcium (mg/100 g) 32.9 ± 0.7 45.9 ± 0.3
a
Prepared from cookies containing 0% (A), 5% (B), 10% (C), 15% (D) Iron (mg/100 g) 0.3 ± 0.1 7.36 ± 0.8
defatted wheat germ flour; casein (E) and protein-free diet (F). Potassium (mg/100 g) 125 ± 1.1 1050 ± 2.0
126 M.U. Arshad et al. / Food Chemistry 102 (2007) 123–128

Table 4
Chemical compositions and mineral content of DFWG-supplemented cookies
Components Defatted wheat germ flour level in cookies (%)
0 5 10 15 20 25
Moisture (%) 8.37 ± 0.10b 8.50 ± 0.09ab 8.66 ± 0.08ab 8.79 ± 0.08ab 8.91 ± 0.08ab 9.02 ± 0.11a
Crude protein (%) 11.8 ± 0.13f 12.7 ± 0.14e 13.6 ± 0.15d 14.5 ± 0.16c 15.4 ± 0.16b 16.2 ± 0.17a
Crude fat (%) 14.15 ± 0.15a 14.09 ± 0.12ab 14.07 ± 0.12ab 14.02 ± 0.11ab 13.98 ± 0.11ab 13.94 ± 0.10ab
Crude fibre (%) 2.33 ± 0.06d 2.56 ± 0.07cd 2.76 ± 0.07bcd 3.01 ± 0.07abc 3.19 ± 0.08ab 3.27 ± 0.08a
Ash (%) 0.76 ± 0.02e 0.95 ± 0.03de 1.13 ± 0.04cd 1.29 ± 0.05bc 1.50 ± 0.05ab 1.690 ± 0.05a
NFE (%) 62.6 ± 1.90a 61.2 ± 1.60b 59.8 ± 1.50c 58.4 ± 1.30d 57.0 ± 1.22e 55.8 ± 1.21f
Calcium (mg/100 g) 50.1 ± 1.20f 50.8 ± 1.30e 51.3 ± 1.30d 52.0 ± 1.40c 52.4 ± 1.45b 53.1 ± 1.46a
Iron (mg/100 g) 1.85 ± 0.05e 2.21 ± 0.06de 2.48 ± 0.06cd 2.71 ± 0.06bc 3.01 ± 0.07ab 3.22 ± 0.07a
Potassium (mg/100 g) 105 ± 3.50f 152 ± 4.05e 196 ± 4.50d 238 ± 5.70c 276 ± 6.05b 306 ± 6.9a
Means in the same row bearing the same letter are not significantly different (p 6 0.05).

All the cookies supplemented with DFWG were found heights and diameters and increased weights; these effects
to be nutritious on the basis of these parameters. This increased with increasing level of replacement of wheat
was because the consumption of about 100 g of each prod- flour with DFWG flour. These results were similar to those
uct formulation would provide more than half of the rec- reported for cookies prepared from wheat–cowpea
ommended daily requirement for protein (25–30 g/day), (McWatters et al., 2003) and wheat–soybean (Shrestha &
as recommended by FAO/WHO (1973) for children aged Noomhorm, 2002) flour blends. There are several views
between 5 and 19 years. This fact suggests that cookies sup- on the mechanisms by which the diameter of cookies (i.e.
plemented with DFWG may be useful as food supplements spread) is reduced when wheat flour is supplemented with
for the alleviation or prevention of protein malnutrition in non-wheat flours. McWatters (1978) reported that rapid
developing countries. partitioning of free water to hydrophilic sites during mixing
Addition of DFWG resulted in an increase in ash values increased dough viscosity, thereby limiting cookie spread.
of cookies up to 1.69% and crude fibre content up to However, it has been suggested that spread ratio is affected
3.27%, while crude fat content was decreased to 13.94% by the competition of ingredients for the available water;
(25% of DFWG level). There were no significant differences flour or any other ingredient, which absorbs water during
in moisture contents of cookies. Supplementation of cook- dough mixing, will decrease it (Fuhr, 1962).
ies with DFWG also significantly (P 6 0.05) increased the
levels of calcium, iron and potassium to 53.1, 3.22 and 3.4. Organoleptic evaluation of cookies
306 mg/100 g, respectively (Table 4).
The effects of DFWG flour on the sensory quality of
3.3. Physical characteristics of cookies cookies are summarized in Table 6. Acceptable cookies
that closely resembled the control (100% wheat flour) cook-
Data on the physical characteristics of cookies are pre- ies were produced from wheat flour containing up to 15%
sented in Table 5. There were no significant (P 6 0.05) dif- DFWG flour. The results of sensory analysis showed that
ferences between the values obtained for spread ratio and scores assigned by the judges for texture were in good
hardness of cookies supplemented with 5–15% DFWG agreement with the measurements derived from the physi-
flour and the control (100% wheat flour) cookies: however, cal (hardness) test. The low overall acceptability of the
these parameters decreased significantly in the case of cookies from blends containing more than 15% DFWG
cookies supplemented with more than 15% DFWG. Blends flour was attributed, by the panellists, to a crumbly texture,
containing more than 15% DFWG flour produced softer a beany flavour and darkening. Colour darkening of cook-
cookies that required less force to compress. In general, ies is attributed to sugar caramelization and the Maillard
cookies made with wheat–DFWG flour blends had reduced reactions between sugars and amino acids (Alobo, 2001).

Table 5
Physical characteristics of DFWG-supplemented cookies
Parameters Defatted wheat germ flour level in cookies (%)
0 5 10 15 20 25
c c b b a
Weight (g) 10.76 ± 0.10 10.79 ± 0.09 11.09 ± 0.12 11.12 ± 0.11 12.46 ± 0.11 12.49 ± 0.13a
Diameter (cm) 9.35 ± 0.08a 9.05 ± 0.10b 7.35 ± 0.08c 6.98 ± 0.06d 4.30 ± 0.04e 4.10 ± 0.04f
Height (cm) 1.30 ± 0.02a 1.27 ± 0.03a 1.25 ± 0.02a 1.00 ± 0.001b 0.75 ± 0.02c 0.72 ± 0.01c
Spread ratio 7.19 ± 0.07a 7.13 ± 0.06a 7.10 ± 0.05a 7.08 ± 0.05a 5.73 ± 0.05b 5.69 ± 0.04b
Hardness (N) 56.51 ± 1.20a 55.96 ± 1.10a 54.68 ± 1.20a 53.91 ± 1.05a 47.89 ± 0.90b 43.56 ± 0.7c
Means in the same row bearing the same letter are not significantly different (p 6 0.05).
 M.U. Arshad et al. / Food Chemistry 102 (2007) 123–128 127

Table 6
Organoleptic characteristics of DFWG-supplemented cookies
Parameters Defatted wheat germ flour level in cookies (%)
0 5 10 15 20 25
Colour 6.80 ± 0.11a 6.30 ± 0.09b 6.40 ± 0.08ab 6.15 ± 0.08b 5.05 ± 0.07c 4.65 ± 0.07c
Flavour 7.50 ± 0.12a 7.50 ± 0.10a 7.45 ± 0.10ab 7.30 ± 0.10b 5.05 ± 0.05c 4.40 ± 0.05d
Texture 6.80 ± 0.10a 6.00 ± 0.08b 5.80 ± 0.07c 5.75 ± 0.07c 3.55 ± 0.05d 3.40 ± 0.05d
OAA 7.65 ± 0.10a 7.35 ± 0.11a 7.50 ± 0.10a 7.65 ± 0.10a 4.85 ± 0.04b 3.90 ± 0.04c
Means in the same row bearing the same letter are not significantly different (p 6 0.05).
A
Overall acceptability.

Table 7
Protein quality parameters of diets containing experimental cookies fed to rats
DietsA Protein quality parametersB
Weight gain (g/10days) Daily feed intake (g) PER NPU (%) BV (%) TD (%)
A 12.50 ± 0.8e 12.30 ± 0.5a 1.11 ± 0.03c 47.4 ± 2.10d 63.2 ± 2.80d 69.5 ± 2.35d
B 27.23 ± 1.0d 12.40 ± 0.5a 1.63 ± 0.05b 50.2 ± 2.40c 65.8 ± 2.80c 75.8 ± 3.10c
C 29.80 ± 1.2c 12.62 ± 0.6a 1.72 ± 0.05b 61.1 ± 2.30b 78.6 ± 2.82b 88.4 ± 3.25b
D 37.42 ± 1.3b 12.31 ± 0.7a 2.78 ± 0.07a 63.3 ± 2.30a 80.6 ± 3.05b 90.6 ± 3.30a
E 40.66 ± 1.3a 12.34 ± 0.7a 2.88 ± 0.08a 65.1 ± 2.81a 85.4 ± 3.25a 93.5 ± 3.30a
PER, protein efficiency ratio; NPU, net protein utilization; BV, biological value; TD, true digestibility.
Means in the same row bearing the same letter are not significantly different (p 6 0.05).
A
Prepared from cookies containing 0% (A), 5% (B), 10% (C) and 15% (D) defatted wheat germ flours; E, casein (control).
B
Values are averages (10 rats per diet) ± standard deviation.

Flavour was found to be a prominent factor in determining higher PER, NPU, BV and TD than did the A (control)
acceptability of DFWG-supplemented cookies. Panellists and B (5% DFWG) diets.
described the cookies containing more than 15% DFWG In short, increases were found in the two directly mea-
flour as having an aftertaste and a beany flavour. This sured parameters, BV and TD, and in the derived value
implies that there is a need to seek preliminary processing for NPU with the increase in DFWG level in cookies. Rats
methods (e.g. heat treatment) to improve the performance fed control diet (100% wheat flour cookies) showed poor
of these flours. Heat treatment (steaming for 30 min at protein quality indices, such as low NPU (47.4%), BV
100 C) was found to be effective for improving the flavour (63.2%) and TD (69.5%). Diets D and E had significantly
of cowpea flour used in cookie-preparation (McWatters, (P 6 0.05) higher values of PER (2.78 and 2.88, respec-
1985). tively) than had the remaining diets. The results suggested
that the nutritional quality of cookies improved with a 10–
3.5. Biological evaluation of protein quality 15% supplementation of DFWG flour. However, cookies
supplemented with DFWG at the 15% level were nutrition-
The cookies containing 5%, 10% and 15% DFWG along ally comparable with diets based on casein. It may be con-
with control (0% DFWG) were found to be the best, based cluded that the differences in protein quality parameters are
on the results of physicochemical and sensory evaluation, because of the essential amino acid content of DFWG pro-
and were subjected to protein quality evaluation through tein which is higher than the control diet containing 100%
a rat bioassay. The results of rats feeding trials, conducted wheat flour.
for the determination of protein quality of cookies, com- The results of this study have shown that highly nutri-
pared with casein, are presented in Table 7. These results tious cookies can be prepared by supplementing wheat
provide the first report of an improvement in the nutritive flour with DFWG flour at 5–15% levels, with increased
value of cookies as a result of DFWG addition. Rats fed on crude protein, calcium, potassium and iron contents. Also,
control diet had the lowest body weight gain (12.34 g), sig- diets based on supplemented cookies containing 15%
nificantly different (p 6 0.05) from the DFWG-supple- DFWG flour were nutritionally comparable with a diet
mented diets (B, C and D) and the casein diet (E). Daily based on casein, indicating that the under-utilized high
feed intakes were not significantly different (p 6 0.05) protein wheat germ could be processed into value-added
among rat groups (Table 7). products and used to combat malnutrition.
The relatively higher essential amino acid balance in
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