International Journal of Gastronomy and Food Science 7 (2017) 20–26

Contents lists available at ScienceDirect

International Journal of Gastronomy and Food Science

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

Scientific Paper

Nutritional, textural and sensory quality of biscuits supplemented with MARK

spinach (Spinacia oleracea L.)☆
⁎
Narsing Rao Galla, Prabhakara Rao Pamidighantam , Balaswamy Karakala, Math
Rudrayya Gurusiddaiah, Satyanarayana Akula
Council of Scientific and Industrial Research-Central Food Technological Research Institute, Resource Centre, Habshiguda, Habshiguda, Uppal Road,
Hyderabad 500 007, Telangana, India

A R T I C L E I N F O A BS T RAC T

Keywords: Spinach (Spinacia oleracea L.) leaves were dried in a cabinet tray dryer at 55 °C and ground to pass through
Spinach powder 220 µm mesh to obtain spinach powder (SP). Fresh spinach leaves yielded 6.5% of SP possessing 28.70%
Nutritional composition protein, 8.8% crude fibre. The powder was rich in essential minerals like calcium (1336 mg/100 g), iron (30 mg/
Antioxidant activity 100 g) and phosphorous (336 mg/100 g). Methanolic extracts (30–180 µL) of SP exhibited antioxidant activity
Biscuits
as assayed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2′-azino-bis(3-ethylbenzthia-
Texture analysis
Sensory quality
zoline-6-sulphonic acid (ABTS) assay and ferric reducing power. Biscuits were prepared using 5%, 10%, and
15% SP and evaluated for their nutritional, textural, sensory quality and sorption behavior. Moisture sorption
isotherm of SP indicated non-hygroscopic nature with an initial moisture content (IMC) of 8.6%, which
equilibrated at 64% relative humidity (RH), whereas, biscuits were observed to be hygroscopic with an IMC of
0.94–1.26%, which equilibrated between 5–30% RH for control sample (CB), 5%, 10% and 15% RH for SP
supplemented biscuits respectively. Textural quality revealed that hardness and breaking strengths increased
with increased addition of SP. Sensory studies of biscuits showed that 5% supplementation of spinach powder
was more acceptable.

Introduction vegetables popular in India were studied. Total chlorophylls ranged

Spinacea oleracea is a leafy vegetable popularly called spinach in
India and consumed in large quantities. Production of spinach world-
wide has reached 20.79 million tons in 2011 and China dominate the
scene with 18.78 million tons (UN FAO., 2011). Spinach leaf is eaten by
people mainly for its characteristic green colour, nutritional content
such as carotenes, vitamin C and minerals such as calcium and iron.
Bioavailability of minerals such as calcium and iron from green leafy
vegetables were assessed and found to be > 25% (Sheetal et al., 2006).
It was observed that the total, insoluble dietary fibre and mineral
contents increased with maturity stages such as tender to mature and
then to a coarse stage of spinach (Punna and Paruchuri, 2004; Khader
and Rama, 1998).
Ascorbic acid and beta-carotene contents in green leafy vegetables
were affected to a greater extent in conventional, pressure cooker and
microwave cookers and deterioration of colour was higher in pressure
cooked samples (Kala and Jamuna, 2004). Effects of blanching and air
drying (60 °C) on colour and vitamin content of different leafy
from 10% to 44% and carotenoids from 42 to 79 µg/g in samples.
Though total chlorophyll levels decreased during drying, chlorophyll-A
was found to be more stable than chlorophyll-B. Retention of carote-
noids was found to be 60–70% in all vegetables except honagone,
where it was only 36% after drying. Beta-carotene and ascorbic acid
were susceptible to oxidation and heat during blanching and drying,
although use of a potassium metabisulphite (KMS) significantly
reduced losses (Premavalli et al., 2001; Negi and Roy, 2000).
Hypolipidemic and hypocholesterolemic effects of fibre from spinach
and mustard leaves were investigated on rats. It was observed that fibre
from spinach helped to significantly lower plasma cholesterol and
glyceride levels and increase the phospholipids (Vadhera et al., 2003).
Phenolic compounds and total antioxidant capacity of salad spinach
was found to be lower than the other vegetables studied (Umma Khair
et al., 2012).
Supplementation of other minor ingredients for enhancement of
nutritional benefits has been practiced the world over. Soy and flax
seed are extensively used for enhancement of protein content and fibre

Peer review under responsibility of AZTI-Tecnalia.

⁎
Corresponding author.
E-mail address: pgp_66@hotmail.com (P.R. Pamidighantam).

http://dx.doi.org/10.1016/j.ijgfs.2016.12.003
Received 27 June 2016; Accepted 19 December 2016
Available online 22 December 2016
1878-450X/ © 2017 AZTI-Tecnalia. Production and hosting by Elsevier B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/).
N.R. Galla et al.
and ω−3 fatty acids respectively in foods. Whole wheat flour was
substituted with 25% and 50% of flaxseed flour (or 12.5% and 25% of
the total flour) in a master mix to evaluate banana bread and peanut
butter and observed that acceptable bread and butter cookies were
produced without affecting characteristics such as flavour, texture, and
mouth feel (Angela et al., 1999). Protein, dietary fibre and ω−3 fatty
acid contents in muffins prepared by incorporating 20% of raw flaxseed
powder were 10.2%, 12.8% and 12.85% respectively and the values
were higher for raw flaxseed powder application, when compared to
roasted flaxseed powder (Chetana et al., 2010). Supplementation of
fenugreek flour in biscuits at 10% level in biscuits was found to be
acceptable with good puffing, increased contents of protein, dietary
fibre, calcium and iron. However expansion in biscuits was found to be
lower than the control (Shalini and Sudesh, 2005). Application of
Murraya koenigii leaf powder at 10% level in cookies resulted in highly
acceptable product in terms of organoleptic, textural and nutritional
qualities (Drisya et al., 2015). Supplementation of defatted mustard
flour at 5%, 10%, 15% and 20% levels in biscuit preparation resulted in
enhanced protein content (2.5 times), fibre and with a reduction in fat
content. Sensory evaluation of biscuits with 15% defatted mustard flour
scored highest for most of the attributes (Tyagi et al., 2007). Biscuits
prepared by supplementation with fenugreek flour at 10% level
resulted in products with enhanced mineral contents of calcium and
iron and they were found to be organoleptically acceptable (Shalini
et al., 2005). Protein and fibre contents in muffins increased with
supplementation of raw flaxseed powder and defatted mustard flour in
the range from 5% to 20% (Chetana et al., 2010; Tyagi et al., 2007).
Reducing the fat content increased the fracture stress of the biscuits,
which indicates that the method provides a useful tool for discriminat-
ing among short-dough biscuits of various compositions (Baltsavias
et al., 1999).
The texture depends on the formulation and the used cooking
regimes (Maache-Rezzoug et al., 1998). Starch is the major structural
element in biscuits, with the fat or sugar playing the structuring role.
The overall quality is largely determined by the type of used fat. In
commercial biscuit samples, significant texture differences were found
as a function of the location of puncturing, as well as the sides of
baking. In biscuits porosity increased with their moisture content, area
of expansion and colour was influenced by relative humidity to a minor
extent (Mandala et al., 2006).
In India green leafy vegetables are popular and they are considered
the most economical vegetables available throughout the year.
However, their application in processed foods for enhancement of
nutritional characteristics has not been explored. The aim of present
study was to prepare a dehydrated leafy vegetable from spinach and
evaluate for its chemical and mineral composition, antioxidant proper-
ties and sorption isotherms. The dehydrated spinach powder was
supplemented in wheat biscuits and their organoleptic, textural,
nutritional and sorption characteristics were investigated.
Materials and methods
Materials
Whole spinach (20 kg) was collected from vegetable market in
Hyderabad, India. Bakery flour (Ushodaya, Hyderabad, India) hydro-
genated fat (Dalda, Bunge India), salt (Annapurna, Hindustan
Unilever), sugar (Reliance Sugar & Chemicals Pvt. Ltd.) and skimmed
milk powder (Sagar, Amul, Gujarat, India) were procured from a
departmental store. Chemicals and solvents used in the study were of
analytical and laboratory grade and purchased from Sd Fine-Chem Ltd.
(Mumbai, India).
Preparation of spinach powder (SP)
Whole spinach leaf was washed with water, soaked in sodium
21
International Journal of Gastronomy and Food Science 7 (2017) 20–26
hypochlorite solution (40 mg/L) for 30 min and dried in a cabinet tray
dryer (Chemida, Mumbai, India) at 55 ± 2 °C for 8 h. The dried
material was ground to powder using a high speed mixer (Sumeet
Domestic Plus, M/s. Sumeet, Nashik, India), passed through BS 72
(220 µm) mesh and dehydrated whole spinach powder (SP) was
obtained. The powder was packed in metallized polyester polyethylene
(MPE) laminate pouches (12 µm metallized polyester, 7.5 µm poly-
ethylene)laminated pouches of size 14×12 cm2 were used for packing
and stored at 4 °C for further chemical analysis and application studies.
DPPH radical scavenging activity
2, 2-Diphenyl-1-picrylhydrzyl (DPPH) radical scavenging activity of
extracts of SP (1 g/100 mL) was measured by using aliquots of 30, 60,
90, 120, 150, 180 µL (Nanjo et al., 1996). Methanolic solution of DPPH
(0.004%) (4 mL) was added and vortexed (Remi, Mumbai, India) for
30 s. The contents were incubated at room temperature (RT) 30 ± 2 °C
for 30 min. The decrease in colour intensity during incubation was
measured in terms of optical density at 517 nm. The percent inhibition
of colour was compared with those of BHT and Trolox equivalent to 5–
30 µg.
ABTS assay
2, 2-Azinobis {3-ethyl-benzothiazoline-6-sulfhonic acid}(ABTS) re-
agent solution (7 mM and the aqueous potassium persulphate
(2.45 mM) were prepared (Re et al., 1999). ABTS stock reagent was
prepared by mixing equal quantities (1:1) of above solutions and
vortexed for 30 s. Further, the stock solution was incubated at RT in
dark for 12–16 h and stored in refrigerator at 3 ± 1 °C. The ABTS stock
solution 1 mL was diluted with 40 mL of distilled water to obtain an
optical density (control) of 0.700 ( ± 0.02) at 734 nm at RT. SP extracts
in methanol (30–180 µL) was added to 3.0 mL of diluted ABTS
solution and incubated at RT in dark for 6 min and OD was recorded
at 734 nm. The % ABTS inhibition was calculated based on control and
sample ODs and compared with BHT and Trolox equivalent to 5–
30 µg.
Ferric reducing power
The ferric reducing power of SP extracts in methanol was measured
according to the reported method in literature (Yildrim et al., 2001).
The extracts (30–180 µL) were added to the mixture of 1 mL methanol
of phosphate buffer in different test tubes. Potassium ferricyanide (1%)
solution (2.5 mL) was added to the test tubes and vortexed for 30 s.
Later, the contents were incubated for 20 min in hot water (50 °C).
After incubation, 2.5 mL of trichloro acetic acid (10%) was added and
centrifuged at 8000 rpm for 10 min. The aliquot of 2.5 mL was mixed
with 2.5 mL of distilled water and 0.5 mL of ferric chloride (0.1%). The
absorbance of the solutions was measured by reading optical density at
700 nm. The ferric reducing power of SP extracts was compared with
those of BHT and Trolox equivalent to 5–30 µg/mL.
Preparation of spinach powder supplemented biscuits (SB)
Control biscuits (CB) and spinach powder supplemented biscuits
(SB) were prepared according to the method reported in the literature
with minor modifications (Tyagi et al., 2007). Spinach powder (SP) was
supplemented at 5%, 10% and 15% levels in bakery flour and dough
was prepared using hydrogenated fat, sugar, salt, ammonium carbo-
nate and water. The dough was sheeted and cut into circular moulds of
3 cm diameter and 1 cm thickness. The moulds were placed in
aluminum trays and exposed to baking temperatures of 190–195 °C
for 15 min. The resultant spinach powder supplemented biscuits (SB)
were cooled to room temperature 30 ± 2 °C. The biscuits were also
prepared without addition of SP, which served as control biscuits (CB).
N.R. Galla et al. International Journal of Gastronomy and Food Science 7 (2017) 20–26

The biscuits were packed in 25 µm polyethylene (PE) and metallized

polyester (2.5 µm) polyethylene (7.5 µm) (MPE) laminated pouches of Fresh spinach leaves (20 kg)
14×12 cm2 size with the unit packs of 10 biscuits and stored at RT (30
± 3 °C).
Washing with water to remove dirt

Physico-chemical composition
Cleaned spinach leaf
The bulk density of SP was measured by noting the volume Sodium hypochlorite treatment (40 ppm) 30 min
occupied by 20 g sample in a 100 mL graduated cylinder. L*, a* and
b* values of SP, CB and SB were analyzed by a colour measuring system
Cutting into smaller bits (3 cm length)
(Hunter Ultra scan, Hunter Laboratory, USA).
Proximate and mineral composition of SP, CB and SB were carried
out using standard methods (Ranganna, 1986). Protein content was Drying at 55 ± 2 °C using tray drier for 8 h 2kg/tray
estimated by using standard micro-Kjeldahl method. In this method a Grinding and passing through BS 72 mesh
sample of 1 g was digested using a mixture of copper sulphate,
potassium sulphate and selenium dioxide in sulphuric acid till all the
Spinach powder (DSP) 1.28 kg (6.40 ± 1.25 %)
organic matter is digested and the nitrogen is converted into ammo-
nium sulphate. The digested sample is made up to a known volume and
ammonia from a small aliquot of the sample (10 mL) is distilled by
Packed in metallised polyester polyethylene pouches
addition of excess of alkali. The distillate is collected into 2% boric acid
solution. Nitrogen content in the distillate is determined by titrating Fig. 1. Flowchart depicting the unit operations for the preparation of spinach powder
with standard 0.1 N hydrochloric acid and the protein content was (SP).
computed using the conversion factor of 6.25 (Pellett and Young,
1980).
The percent carbohydrate content and the energy value were Table 1
calculated by difference using the following equations: Antioxidant activity of methanol extracts of spinach powder (SP), BHT and troloxa.

% Carbohydrates = [100–(Moisture+Total ash + Protein + Fiber+Fat)] Quantity (µg/ DPPH (% ABTS (% FRP
mL) inhibition) inhibition) (OD)
(1)
30 22.56 47.37 0.051
Energy (kcal/100g) = 4 (% Protein + % Carbohydrate) + 9(% Fat) (2)
60 23.94 48.08 0.075
90 24.69 56.32 0.095
120 26.70 61.60 0.116
150 28.18 63.85 0.127
Mineral content 180 36.71 68.34 0.144

BHT
Minerals such as calcium, iron and phosphorous in SP, CB and SB
30 45.31 35.79 0.188
were determined by preparing solutions of total ash in 6 M hydro- 60 48.09 41.97 0.422
chloric acid. Calcium content was estimated by precipitating it as 90 60.35 50.28 0.505
calcium oxalate and titrating with standard potassium permanganate 120 62.87 58.49 0.631
solution; iron content was estimated using colourimetric method using 150 85.15 62.79 0.721
180 69.08 70.36 0.814
UV–Visible spectrophotometer (Shimadzu, UV-160A model) at
480 nm (AOAC, 1995). Phosphorous was analyzed by developing Trolox
colour using ammonium molybdate and 2, amino −6, naphthol 5 37.12 56.88 0.047
sulphonic acid. The blue colour developed was read at 650 nm in 10 42.93 68.92 0.056
15 50.88 73.07 0.088
UV–Visible spectrophotometer (Shimadzu, UV-160A model) and ex-
20 59.01 86.70 0.103
pressed as phosphorus mg/100 g. 25 68.20 99.14 0.146
30 84.62 – 0.152
Equilibrium moisture content-relative humidity (EMC-RH) studies a
Values are average of triplicate analysis with ± SD (0.001–1.5).
DPPH: 2,2-diphenyl-1-picrylhydrazyl 1,1-diphenyl-2-picrylhydrazyl radical.
The sorption behavior of SP, CB and SB were determined by ABTS: 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid.
exposing the samples (5 g) to different relative humidity (RH) ranging FRP: Ferric reducing power.
from 10%, 30%, 50%, 70%, 90% and 100% maintained by using OD: Optical density.
appropriate concentrations of sulphuric acid in glass desiccators at BHT: Butylated hydroxyl toluene.
Trolox: (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid).
room temperature (RT) 30 ± 2 °C. The samples were observed visually
for change in colour, lump formation and mold growth during the
study. The samples were weighed at regular intervals till they attained
where A is the weight of experimental sample and B is its initial
constant weight. Moisture sorption isotherms were plotted for the
moisture content (IMC)
equilibrium moisture content (EMC) and the corresponding equili-
brium relative humidity relative humidity (ERH) (Ranganna, 1986). D−C
Equilibrium Moisture Content (EMC%), = × 100
Lump formation and mold growth during the study was noted, D (4)
calculated the moisture content at this stage and expressed as critical where D is the weight of sample after equilibration
moisture content (CMC). The dry solids and EMC were calculated using
following equations:
Texture measurements
A × 100 − B
Dry solids of the sample (C) =
100 (3) Texture analysis of biscuits was carried out using Brookfield texture

22
N.R. Galla et al. International Journal of Gastronomy and Food Science 7 (2017) 20–26

Bakery flour (4 kg)

Blending of hydrogenated fat (2 kg), salt (32 g), sugar (1 kg), water (1000 g), milk powder (400 g),

ammonium carbonate (60 g) in a planetary mixer, blending of dehydrated spinach powder replacing

5, 10 and 15% levels of bakery flour

Bakery dough

Conditioning 30 min

Conditioned dough

Kneading forming of sheet and cutting of biscuit

Baking at 190-195 C for 15 min.

________________________________________________________

Control biscuits Biscuits with 5% Biscuits with 10% Biscuits with 15%
(CB) SP (SB-5) SP (SB-10) PS (SB-15)
_________________________________________________________

Packaging in metallised polyester polyethylene pouches

10 biscuits per pack (average total yield 6.4 kg)

Fig. 2. Flowchart of the unit operations for the preparation of spinach supplemented biscuits.

analyzer (Model CT3, Middleboro, USA) and breaking strength and Results and discussion
compression tests were performed using test probes namely TA-7 and
TA-25 respectively. Yield and bulk density of spinach powder

Unit operations involved in the preparation of spinach powder are

Sensory evaluation of biscuits
The CB, SB-5%, SB-10% and SB-15% were subjected to sensory
evaluation of ten trained panelist from CSIR-CFTRI-RC, Hyderabad,
India on their familiarity with the biscuits were used for the evaluation.
The parameters evaluated such as appearance, colour, flavour, aroma,
taste, texture and overall acceptability. The coded samples were served
in clean white plastic plates at RT in individual booths with adequate
florescent lights. Sample presentation to the panelists was at random
and one at a time. They were provided samples to eat and check how
much they liked or disliked each one and rate them as such. The
panelists were given enough water to rinse their mouths between each
serve. The sensory evaluations were collected on 9-point Hedonic scale.
The panelists scored for different parameters with a maximum score of
9 for like extremely, 8-like very good, 7-like good, 6-like moderately, 5-
neither like nor dislike, 4-dislike moderately, 3- dislike fairly, 2-dislike
very much and 1 for dislike extremely (Amerine et al., 1965).
Statistical analysis
Nutritional composition, sorption isotherm studies, textural analy-
sis and sensory quality of biscuits were carried out in triplicate and
mean values with standard deviation (SD) were computed by using
Microsoft Excel, 2007.
presented in Fig. 1. Processing of fresh spinach leaves on dehydration
yielded 6.5 ± 1.25% of spinach powder (SP). with an average bulk
density of 0.6634 ± 0.0014 g/cc.
Antioxidant activity of spinach powder
Antioxidant activity of spinach powder could be established by all
the three methods employed, i.e. DPPH radical scavenging activity
ABTS assay and ferric reducing power (Table 1). In all the methods
increasing concentration of SP resulted in higher activity. The activity
was more pronounced in the method of ABTS and FRP as seen from the
data. It was observed that inhibition of DPPH and ABTS radicals was to
the extent of 36.75% and 68.34% with 180 µg/mL of extract respec-
tively. In FRP, the increase in optical density was much higher and the
OD of 0.051 at 30 µL increased to 0.144 for a concentration of 180 µg/
mL. However, the activity was much lower when compared to standard
antioxidants such as BHT or Trolox. Earlier studies on antioxidant
activity of leafy vegetables showed maximum DPPH scavenging activity
and reducing power was exhibited by Murraya koenigii when com-
pared with Amaranthus sp., Centella asiatica and Trigonella foenum
graecum (Sheetal and Jamuna, 2009). The ethanolic extracts of leafy
vegetables namely cabbage (Brassica oleraceavar. capitata), coriander
leaves (Coriandrum sativum), hongone (Alternanthera sessilis), and
spinach (Spinacia oleracea) analyzed for antioxidant activity in heated

23
N.R. Galla et al. International Journal of Gastronomy and Food Science 7 (2017) 20–26

protein content by 2.5 times along with a reduction in fat and an

increase in fibre contents. However, the sample with 15% defatted
mustard flour was the most acceptable product with similar sensory
and texture attributes of control (Tyagi et al., 2007).
Colour values of dehydrated spinach showed greenness indicated by
a negative value for a*(−3.36) and the biscuits supplemented with SP
could not retain the greenness during baking. Other Hunter colour
parameters varied widely on increasing addition of spinach. The
changes in Hunter L*, a* and b* and darkening of products may be
due to browning of spinach carbohydrates during baking (Table 2).
Addition of spirulina powder, whole wheat and sorghum flours in
biscuits increased its protein, fibre as well as antioxidant potential
however, high levels of fortification ( > 7% spirulina powder and > 30%
sorghum flour) adversely affected the colour, textural and sensory
attributes of biscuits (Parul et al., 2015).

Experimental sorption isotherms

(CB) (SB-5%) (SB-10%) (SB-15%) Experimental sorption isotherms of SP, SB-5%, SB-10% and SB-
15% were presented in Fig. 3. These studies indicated that the PP was
non-hygroscopic in nature but biscuits supplemented with SP have
60
shown to be hygroscopic in nature. Initial moisture contents (IMC) and
PP CB 5% PSB 10% PSB 15% PSB critical moisture content (CMC) of SP were 8.63% and 17.61%, which
% Equilibrium moisture content

50
equilibrated at 64% and 69% % RH respectively. The CMC of the above
samples were 17.61%, 8.45%, 9.6% and 10.2%, which equilibrated in
40
the range 70–81% RHs. Increasing addition of PP has further increased
the stability towards humidity. The present study indicated that the PP
30
was non- hygroscopic and biscuits were slightly hygroscopic in nature.
Hence, PE or MPE pouches with high moisture barrier properties for
20
storage at RT may be recommended. Biscuits prepared using defatted
protein powder with a CMC of 20% were reported in literature
10
(Balaswamy et al., 2009). In our earlier studies on seed protein
concentrates from sterculia and bael, critical moisture contents of
0
10 30 50 70 90 100 9.2%, 8.76%, which equilibrated at 72% and 68% RH respectively were
% Relative humidity reported (Narsing Rao and Rao, 2010; Narsing Rao et al., 2011).

Fig. 3. (A) Photographs of control biscuits (CB) and spinach powder supplemented
biscuits (SB) (5–15% levels); (B)Sorption isotherms of dehydrated spinach powder (SP),
control biscuits (CB), PP supplemented biscuits 5–15% (SB-5%, SB-10% and SB-15%).
refined sunflower and groundnut oils showed enhanced stability
against peroxide formation for 4 weeks (Shyamala et al., 2005). The
reducing power of the leafy vegetables followed the order: spinach <
cabbage < coriander leaves < hongone leaves.
Texture measurements
Texture analysis of biscuits using Brookfield texture analyzer for
parameters such as breaking strength and compression tests indicated
addition of SP has affected the textural quality. Hardness increased
Table 2
Nutritional composition and colour characteristics of spinach powder (SP), control
biscuits (CB) and spinach powder supplemented biscuits (SB)a.

Nutritional composition of SP and SB Biscuits

Photographs of fresh spinach leaves and the dehydrated spinach Parameters, % SP CB SB-5 SB-10 SB-15
powder are presented in Fig. 1. Flow chart depicting the unit opera- Moisture 8.60 0.94 1.12 1.22 1.26
tions for the preparation of spinach supplemented biscuits is presented Total ash 18.81 2.06 3.45 4.87 5.97
in Fig. 2. The photographs of biscuit products with varying concentra- Crude fat 5.69 28.09 26.40 25.10 24.76
tions of spinach are presented in Fig. 3. Crude protein 27.8 5.88 7.54 8.88 10.79
Crude fibre 8.82 0.89 1.22 2.31 2.87
Nutritional composition of SP and SB-5–15% are presented in
Carbohydrates 30.28 62.14 60.27 57.62 54.35
Table 2. Spinach powder possessed good quantities of protein 27.8%, Energy (kcal/100 g) 294.33 524.89 508.84 492.20 483.40
fibre 8.8% along with minerals such as calcium 1336 mg/100 g, iron
30 mg/100 g and phosphorous 336 mg/100 g. Increasing addition of Minerals (mg/100 g)
spinach (5–15%) has shown good enhancement in protein, minerals Calcium 1336 98.66 203.75 337.89 460.18
Iron 30 2.85 3.74 5.64 7.89
and fibre in biscuits when compared to control. Filip and Vidrih (2015)
Phosphorus 336 54.44 77.42 101.47 132.23
produced high-protein pasta using 40% of pea protein isolate and 3%
dry spinach powder. The pasta products respectively possessed 36.4 g Colour characteristics
and 39.6 g protein per 100 g of dry matter when compared to 10% in L* 55.36 66.15 47.96 42.07 40.74
a* −3.35 11.86 1.52 0.40 0.61
plain durum wheat pasta and would yield energy of 386 and 366 Kcal
b* 13.91 33.95 14.26 5.58 4.42
per 100 g respectively, and a reduction in carbohydrates (44.6, 40.4%)
from 77% in wheat pasta. Incorporation of defatted mustard flour at a
Values are average of triplicate analysis with ± SD (0.001–1.5).
5%, 10%, 15% and 20% in biscuit preparation resulted in enhanced

24
N.R. Galla et al. International Journal of Gastronomy and Food Science 7 (2017) 20–26

Table 3 Acknowledgements
Texture analysis and sensory quality of spinach powder supplemented biscuits.
The authors thank the Director, CSIR-CFTRI, Mysore and CFTRI-
Parameters CB SB-5% SB-10% SB-15%
RC staff for their support and encouragement in carrying out the work.
Texture analysis
Hardness (g) 2138 ± 4.04 2395 ± 5.29 2498 ± 7.00 3403 ± 6.51 References
Breaking strength 90 ± 3.06 808 ± 8.08 360 ± 8.14 318 ± 6.51
(Load in g)
Agu, H.O., Ayo, J.A., Paul, A.M., Folorunsho, F., 2007. Quality characteristics of biscuits
made from wheat flour. Niger. Food J. 25 (2), 19–27.
Sensory scores
Amerine, M.A., Pangborn, R.M., Roessler, E.B., 1965. Principles of Sensory Evaluation of
Appearance 8.71 ± 0.62 7.67 ± 0.78 6.68 ± 1.21 6.60 ± 1.21
Food. Academic Press, New York.
Colour 8.33 ± 0.65 7.42 ± 1.00 6.63 ± 1.46 6.50 ± 1.51 Angela, F., James, H., Julie, A.A., Sheila, E.S., 1999. Incorporation of flaxseed into a
Flavour 8.00 ± 1.13 7.33 ± 0.89 6.54 ± 1.20 6.33 ± 1.60 master mix and evaluation of banana bread and peanut butter cookies. J. Am. Diet.
Texture 8.17 ± 0.83 7.92 ± 0.67 7.13 ± 0.91 6.83 ± 1.19 Asso. 99 (9), 29.
Taste 8.25 ± 0.87 7.42 ± 0.90 6.21 ± 1.50 5.92 ± 1.62 UN Food and Agriculture Organization, 2011. Production of Spinach by countries. FAO,
Overall acceptability 8.08 ± 1.00 7.42 ± 0.90 6.54 ± 1.41 6.00 ± 1.65 Geneva.
AOAC, 1995. Official Methods of Analysis 16th edn. Association of official Analytical
a
Values are average of triplicate analysis with ± SD. Chemists, Washington, DC.
CB: Control biscuits (without supplementation of SP). Balaswamy, K., Prabhakara Rao, P.G., Narsing Rao, G., Rao, D.G., Jyothirmayi, T., 2009.
Physico-chemical composition & functional properties of roes from some fresh
SB: Spinach powder supplemented biscuits.
water fish species & their application in some foods. Elec. J. Env. Agric. Food Chem.
8, 806–812.
Baltsavias, A., Jurgens, A., Van, Vliet, T., 1999. Properties of shortdough biscuits in
from upon addition of spinach at 5–15% levels (Table 3). A similar relation to structure. J. Cereal Sci. 29, 245–255.
trend was noticed in cookies where hardness increased with increased Chetana, R., Sudha, M.L., Khyrunnisa, B., Ramasarma, P.R., 2010. Nutritional
addition of dehydrated Murraya koenigii powder (Drisya et al., 2015). characteristics of linseed/flaxseed (Linum usitatissimum) and its application in
muffin making. J. Texture Stud. 41 (4), 563–578.
However, breaking strength showed a higher value of load (808 g) for Drisya, C.R., Swetha, B.G., Velu, V., Indrani, D., Singh, R.P., 2015. Effect of dried
5% application when compared to control, which decreased to 360 and Murraya koenigii leaves on nutritional, textural and sensory characteristics of
318 g respectively when 10, 15% PP were incorporated. The higher cookies. J. Food Sci. Technol. 2 (1), 500–506.
Filip, S., Vidrih, R., 2015. Amino acid composition of. Food Technol. Biotechnol. 53 (3),
levels of spinach might have added higher fibre resulting in lower 298–306.
binding of carbohydrates and further lesser breaking strengths. Though Kala, A., Jamuna, P., 2004. Nutrient composition and sensory profile of differently
firmness tends to increase with fibre content in biscuits, there were cooked green leafy vegetables. Int. J. Food Prop. 7 (3), 659–669.
Khader, V., Rama, S., 1998. Selected mineral content of common leafy vegetables
some exceptions (Piteira et al., 2006). Hardness of the high fibre and consumed in India at different stages of maturity. Plant Food Hum. Nutr. 53 (1),
high protein biscuits by incorporation of spirulina and sorghum flours 71–81.
varied between 1545.14 and 2298.14 g and fracturability of the high Maache-Rezzoug, Z., Bouvier, J.M., Allaf, K., Patras, C., 1998. Effect of principal
ingredients on rheological behaviour of biscuit dough and on quality of biscuits. J.
fibre and high protein biscuit ranged between 0.341 and 0.644 mm
Food Eng. 35, 23–42.
indicating high content of fibre enhanced its fracturability (Parul et al., Mandala, I.G., Ioannou, C.A., Kostaropoulos, A.E., 2006. Textural attributes of
2015). In biscuits, application of breadfruit flour at 10% level showed commercial biscuits. Effect of relative humidity on their quality. Int. J. Food Sci.
Technol. 41, 782–789.
an increase of breaking strength from 139.8 to 299.2 g (Agu et al.,
Nanjo, F., Goto, K., Seto, R., Suzuki, M., Sakai, M., Hara, Y., 1996. Scavenging effects of
2007). These parameters indicate that crispness increased with in- tea catechins and their derivatives on 1, 1, -diphenyl-2picrylhydrazyl radical. Free
crease of SP in biscuits. However, the addition of SP at 5% level did not Radic. Biol. Med. 21, 895–902.
affect hardness though breaking strength increased. At higher levels of Narsing Rao, G., Rao, D.G., 2010. Chemical and functional characterization of gum
karaya (Sterculia urens L.) seed meal. Food Hydrocolloid. 24 (5), 479–485.
SP, breaking strength was comparable with that of control, however Narsing Rao, G., Balaswamy, K., Prabhakarara Rao, P.G., Rao, D.G., Satyanarayana, A.,
overall acceptability scores decreased. 2011. Nutritional quality, fatty acids, amino acids and functional characteristics of
bael (Aegle marmelos l.) seed protein concentrate. J. Food Drug Anal. 19 (4),
528–535.
Negi, P.S., Roy, S.K., 2000. Effect of blanching and drying methods on beta-carotene,
ascorbic acid and chlorophyll retention of leafy vegetables. LWT Food Sci. Technol.
Sensory analysis 33 (4), 295–298.
Parul, S., Rakhi, S., Alok, J., Prasad, R., Anuj Kumar, G., 2015. Optimization of a process
The data related to sensory evaluation was presented in Table 3. In for high fibre and high protein biscuit. J. Food Sci. Technol. 52 (3), 1394–1403.
Pellett, L.P., Young, V.R., 1980. Nutritional evaluation of protein foods. UN: Univ. Food
case of all attributes SB-5% scored almost nearer to with the control Nutr. Bull. (Suppl. 4), (Tokyo).
samples, when compared to SB-10 and SB-15%. Darkness in the Piteira, M.F., Maia, J.M., Raymundo, A., Sousa, I., 2006. Extensional flow behaviour of
samples increased with increase in PP content of biscuits. Flavour natural fibre-filled dough and its relationship with structure and properties. J. Non-
Newton. Fluid Mech. 137, 72–80.
decreased as SP increased and bitterness was noticed as after taste in
Premavalli, K.S., Majumda, T.K., Madhura, C.V., 2001. Processing effect on colour and
SB-15%. Overall scores indicated highly acceptable for SB-5%, 10% and vitamins of green leafy vegetables. J. Food Sci. Technol. 38 (1), 79–81.
15% and comparable to control (CB). It was observed that supplemen- Punna, R., Paruchuri, R.U., 2004. Effect of maturity and processing on total, insoluble
and soluble dietary fiber contents of Indian green leafy vegetables. Int. J. Food Sci.
tation of Murraya koenigii leaf powder at 10% level was found to be
Nutr. 55 (7), 561–567.
acceptable in cookies with an pleasant curry leaf flavour and quality Ranganna, S., 1986. Hand Book of Analysis & Quality Control for Fruits & Vegetable
(Drisya, 2015). Products 2nd edn.. Tata McGraw-Hill Publishing Company Limited, New Delhi.
Re, R., Pellegrini, N., Proteggente, A., Yang, M., Rice-Evans, C., 1999. Antioxidant
activity applying an improved ABTS radical cation decolourization assay. Free Radic.
Biol. Med. 26 (9–10), 1231–1237.
Shalini, H., Sudesh, J., 2005. Organoleptic and nutritional evaluation of wheat biscuits
Conclusions supplemented with untreated and treated fenugreek flour. Food Chem. 90 (3),
427–435.
It has been successfully demonstrated that dehydrated spinach Sheetal, G., Jyothi, L., Jamuna, P., 2006. In vitro bioavailability of calcium and iron from
selected green leafy vegetables. J. Sci. Food Agric. 86 (13), 2147–2152.
powder could be safely employed as a protein, mineral and fibre source Sheetal, G., Jamuna, P., 2009. Studies on Indian green leafy vegetables for their
in biscuits at 5% level. Protein content up to 28.2% was achieved by the antioxidant activity. Plant Foods Hum. Nutr. 64 (1), 39–45.
addition of 5% of dehydrated spinach. A reduction in fat content in the Shyamala, B.N., Sheetal, G., Jyothi Lakshmi, A., Jamuna, P., 2005. Leafy vegetable
extracts—antioxidant activity and effect on storage stability of heated oils. Innov.
biscuits can be highlighted with increased addition of spinach. The Food Sci. Emerg. Technol. 6 (2), 239–245.
spinach incorporated biscuits can be used as an alternative source to Tyagi, S.K., Manikantan, M.R., Harinder Singh, O., Gurlen, K., 2007. Effect of mustard
meet the fibre requirements. flour incorporation on nutritional, textural and organoleptic characteristics of

25
N.R. Galla et al. International Journal of Gastronomy and Food Science 7 (2017) 20–26

biscuits. J. Food Eng. 80 (4), 1043–1050.
Umma Khair, S.K., Shinya, O., Emiko, Y., Yoshiya, M., 2012. Phenolic acids, flavonoids
and total antioxidant capacity of selected leafy vegetables. J. Funct. Food 4 (4),
979–987.
Vadhera, S., Sangeeta, W., Giridhar, S., 2003. Hypocholesterolemic/hypolipidemic
effects of fibre from leafy vegetables (spinach and mustard). J. Food Sci. Technol. 40
(5), 531–533.
Yildrim, A., Mavi, A., Kara, A.A., 2001. Determination of antioxidant and antimicrobial
activities of Rumex cripsus L. extracts. J. Agric. Food Chem. 49, 4083–4089.

26