LWT - Food Science and Technology 98 (2018) 77–84

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LWT - Food Science and Technology

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Effect of chia seed on glycemic response, texture, and sensory properties of T

Chinese steamed bread
Fan Zhu∗, Coline Chan
School of Chemical Sciences, University of Auckland, Private Bag 92019, Auckland 1142, New Zealand

A R T I C LE I N FO A B S T R A C T

Keywords: Chia (Salvia hispanica) seed is a source of diverse bioactive components with various potential health effects.
Salvia hispanica Chinese steamed bread (CSB) is a traditional staple among Asian population. In this report, intact white chia
In vitro starch digestibility seeds replaced wheat flour for northern-style CSB formulation at various levels up to 300 g/kg. The impact of
Rheology seed replacement on in vitro starch digestion, expected glycemic index (eGI), expected glycemic load (eGL),
Glycemic index
volumetric properties, colour, texture, and sensory quality of the CSB were studied. Chia seed addition reduced
Functional food
the eGI and eGL of CSB by up to 25% and 48%, respectively. Chia seeds increased the hardness of CSB and
reduced the specific volume of CSB by up to 211% and 27%, respectively. Sensory analysis showed that the
overall acceptance of CSB was not affected by chia seed addition. It may be concluded that intact chia seed
addition up to 300 g/kg can nutritionally enhance CSB without compromising the eating quality.

1. Introduction glucoronopyranosy residues branching at O-2 position of some of the β-

Among the “novel” and “healthy” functional food ingredients, chia
(Salvia hispanica L.) seed has gained increasing interest due to the un-
ique nutritional properties (de Falco, Amato, & Lanzotti, 2017). Chia of
the mint family Lamiaceae is native to central and southern Mexico and
Guatemala (Muñoz, Cobos, Diaz, & Aguilera, 2013; de Falco et al.,
2017). It was an important crop in the pre-Columbian time. Chia seeds
are oval with a very small size of ∼1–2 mm, and are mottle-coloured
with white, gray, brown, or black (Muñoz et al., 2013; de Falco et al.,
2017). The seed is consisted of proteins (190–230 g/kg), lipids
(300–330 g/kg), carbohydrates (260–410 g/kg), dietary fibre
(180–300 g/kg), and ash (40–50 g/kg) along with polyphenols derived
from caffeic acid, vitamins, and others (Muñoz et al., 2013; de Falco
et al., 2017). It contains more protein than any grain and has a higher
fibre content compared to flaxseed, quinoa, and amaranth. The seed has
been found to contain the highest known percentage of α-linolenic fatty
acid (ALA) of any plant source which can amount up to over 600 g/kg
of the total lipids (de Falco et al., 2017). In particular, when coming
into contact with water, chia seed excretes a mucilage of polysaccharide
(50–60 g/kg) with a molecular weight of 0.8–2 × 106 (Lin, Daniel, &
Whistler, 1994; Muñoz, Cobos, Diaz, & Aguilera, 2012). The poly-
saccharide was tentatively identified to have a basic unit of tetra-
saccharide. The main backbone is of (l→4)-β-D-xylopyranosyl-(l→4)-α-
D-glucopyranosyl-(l→4)-β-D-xylopyranosyl units with 4-O-methyl-α-D-
D-xylopyranosyl residues (Lin et al., 1994). The mucilage has a high
water holding capacity and can act as an active hydrocolloid with a
high water holding capacity, potentially improving dough properties of
breadmaking (Iglesias-Puig & Haros, 2013). Because of the attractive
nutritional and functional profile and also the positive opinions from
government agency (e.g., European Food Safety Authority), intact chia
seeds and flour have been readily added to Western bakery products
such as bread (Steffolani, Martinez, León, & Gómez, 2015).
Chinese steamed bread (CSB) (also called mantou or steamed bun) is
a staple, being widely consumed in China and among Asian population.
The basic ingredients of CSB are wheat flour, water, and yeast (Zhu,
2014). The ingredients are mixed to form a dough for fermentation. The
fermented dough is steamed to make CSB. There is a great opportunity
to develop “healthy” CSB with added functional ingredients (Zhu,
2014). For example, CSB of refined wheat flour, which is the most
consumed CSB form in China, has a high glycemic index (GI) (Jiao
et al., 2003). The large population of diabetic people in China may
benefit from eating CSB with a lower GI and a lower glycemic load (GL)
(Jenkins et al., 2002). A number of “healthy” ingredients such as black
tea have been incorporated into CSB formulation (Zhu, Sakulnak, &
Wang, 2016). Chia seed, as a novel functional food, remains to be used
for CSB formulation with enhanced nutritional benefits. The very di-
verse climate and ecological regions of China would be suitable for the
large scale cultivation of chia. In fact, chia is being developed in China

∗
Corresponding author.
E-mail address: fzhu5@yahoo.com (F. Zhu).

https://doi.org/10.1016/j.lwt.2018.08.016
Received 31 March 2018; Received in revised form 6 August 2018; Accepted 7 August 2018
Available online 07 August 2018
0023-6438/ © 2018 Elsevier Ltd. All rights reserved.
F. Zhu, C. Chan
for potential large scale cultivation (Gao, Tao, Ning, Rong, & Zhang,
2017).
In this study, intact chia seeds replaced wheat flour up to 300 g/kg
for CSB making. The impact on texture, shelf life, in vitro starch di-
gestibility, expected glycemic index (eGI), expected glycemic load
(eGL), and sensory properties of CSB were studied. The pasting and gel
textural properties of wheat flour affected by chia addition were also
tested. The impact of ground chia seed flour on the quality attributes of
CSB was also studied, and compared with that of intact chia seed to
assess the practical application of different chia products available from
the market.
2. Materials and methods
2.1. Materials
All-purpose wheat flour (Pams Food, Auckland, New Zealand),
whole white chia seeds (Macro, Bella Vista, Australia), ground chia
seeds (Healtheries, Auckland, New Zealand), and yeast (Edmons,
Auckland, New Zealand) were obtained from a local supermarket
(Countdown, Auckland, New Zealand). The proximate analysis
(moisture, 131 g/kg; ash, 7 g/kg; protein, 111 g/kg, and lipid, 1.3 g/kg)
of the wheat flour was conducted using the Approved AACCI Methods
44–40.01, 08–03.01, 46–12.01, and 30–25.01 (AACC International,
2000). The total starch content of the flour was 716 g/kg as determined
by a Megazyme total starch kit (Wicklow, Ireland). The nutritional
composition of the intact chia seeds and ground chia seeds according to
the suppliers was summarised in Supplementary Table 1. The two ap-
peared to be similar. The contents of dietary fibre, lipids, and protein of
the intact chia seeds were 380 g/kg, 351 g/kg and 180 g/kg, respec-
tively. The substitution levels of chia seeds in wheat flour were 25 g/kg,
50 g/kg, 75 g/kg, 100 g/kg, 150 g/kg, 200 g/kg, and 300 g/kg in dif-
ferent studies. Preliminary study showed that a higher percentage of
chia seed addition (> 300 g/kg) drastically decreased the eating quality
of CSB.
2.2. Analysis of wheat flour systems
2.2.1. Pasting analysis
The pasting properties of the flour samples were analyzed using an
MCR 301 Rheometer equipped with a starch cell (Anton Paar, Graz,
Austria). The procedure followed a previous report (Li & Zhu, 2017).
Wheat flour or flour-chia seed mixtures (3.0 g, db) were dispersed in
20 mL of water before subjecting to a heating and cooling cycle of
26 min. Peak viscosity (PV), hot paste viscosity (HPV), and cool paste
viscosity (CPV) were noted. Breakdown (BD = PV − HPV) and setback
(SB = CPV − HPV) were calculated (Li & Zhu, 2017).
2.2.2. Gel textural analysis
After pasting, the paste (5 mL) was transferred to a canister with a
screw cap. The canister was subjected to centrifugation (1000 × g,
5 min) to remove air bubbles and for a flat surface. The gel was stored at
4 °C for 24 h before texture profile analysis (TPA) by using a TA-XT2i
plus Texture Analyser (Stable Micro Systems Ltd, Godalming, UK) as
described previously (Li & Zhu, 2017). Hardness and adhesiveness of
the gels were recorded.
2.3. Production and analysis of Chinese steamed bread (CSB)
2.3.1. Production of CSB
The production of CSB followed a previous method (Jiang, Cong,
Yan, Kumar, & Du, 2010) with some modifications. Briefly, the dried
yeast (0.8 g) was dispersed in water (40 °C, 50 mL) for 10 min before
mixing with wheat flour (100 g). For the wheat flour-chia seed mixture
samples, the optimal amount of added water for dough formation was
determined by trial and error approach. The ingredients were mixed
78
LWT - Food Science and Technology 98 (2018) 77–84
and hand-kneaded for 3 min to form a dough before proofing at 37 °C
for 40 min (relative humidity of 80%). The proofed dough was then
hand-kneaded for 1 min, shaped, and proofed for another 15 min at
37 °C. The fully proofed dough was then steamed in a steaming chamber
(100 °C) for 20 min, and cooled for 30 min at room temperature before
subsequent analysis.
2.3.2. In vitro starch digestibility and expected glycemic index of CSB
The in vitro starch digestibility of CSB samples was measured fol-
lowing the method of Englyst, Kingman, and Cummings (1992). Briefly,
a piece of CSB (1.8 g) was made into a slurry (100 g/L) before the ad-
dition of sodium acetate buffer (10 mL, 0.1 M, pH 5.2) and a fresh en-
zyme mixture (5 mL) consisting of pancreatin (P1750, 500 μkat/g,
Sigma-Aldrich Chemical Co., St. Louis, USA), invertase (EC 3.2.1.26,
≥5 μkat/mg, Sigma-Aldrich Chemical Co., St. Louis, USA) and amy-
loglucosidase (3.3 μkat/mL, Megazyme, Wicklow, Ireland) at a ratio of
54:6:4 (volume basis). The sample was placed in a shaking water bath
(37 °C) for 2 h. An aliquot (0.1 mL) of sample was taken every 20 min
and 4 mL of 80% (by volume) ethanol was added to stop the hydrolysis.
The glucose concentration in the sample was determined by a Mega-
zyme D-glucose (glucose oxidase/peroxidase; GODPOD) assay kit
(Megazyme, Wicklow, Ireland). Rapidly digestible starch (RDS), slowly
digestible starch (SDS), and resistant starch (RS) fractions were quan-
tified as the percentage of total dry matter which was described by
Englyst et al. (1992).
The non-linear first order equation, C = C∞ (1−e-kt) was used to
describe the kinetics of the hydrolysis of the sample (Goñi, Garcia-
Alonso, & Saura-Calixto, 1997). C is the percentage of starch hydrolysed
at time t; C∞ is the percentage of starch hydrolysed at 120 min; k is the
kinetic constant; and t is time. The area under the hydrolysis curve
(AUC) was determined through curve fitting by Origin (version 8.5,
OriginLab Corporation, Northampton, MA, USA). The hydrolysis index
(HI) was calculated by dividing AUC of CSB sample with the AUC of
white bread (Homebrand, Countdown Supermarket, Auckland, New
Zealand) (as a reference sample). The HI of the white bread was set as
100% as a reference. The expected glycemic index (eGI) was calculated
from the equation eGI = 39.71 + 0.549HI (Goñi et al., 1997). The
expected glycemic load (eGL) per 100 g of CSB was calculated by
multiplying the eGI with the total starch percentage.
2.3.3. Specific volume and spread ratio of CSB
The specific volume was determined by Volscan VSP600 (Stable
Micro Systems Ltd, Godalming, UK). The spread ratio was derived from
width divided by height of CSB.
2.3.4. Water content and water activity of CSB
The AquaLab 4 TE water activity meter (Decagon Devices Inc.,
Pullman, Washington, USA) was used to measure the water activity of
CSB crumb. The Infrared Moisture Analyser MA35 (Sartorius,
Gottingen, Germany) was used to measure the moisture content of
crushed CSB crumb.
2.3.5. Colour of CSB
The colour of the crust and crumb of CSB was quantified by a CR-
300 chroma-meter (Minolta Camera Co., Osaka, Japan), following the
method of Zhu, Cai, Sun, and Corke (2008). Six different points on the
surface of the samples were randomly selected for the colour mea-
surement. Data were reported in the form of CIE L*, a*, and b* colour
space. The total colour difference (ΔE) against the control CSB without
chia seeds was calculated using the equation:
ΔE = (ΔL2 + Δa2 + Δb2)1/2
2.3.6. Texture profile analysis of CSB
Texture profile analysis (TPA) was conducted on CSB samples using
a TA-XT2i plus Texture Analyser equipped with an aluminium
F. Zhu, C. Chan LWT - Food Science and Technology 98 (2018) 77–84

cylindrical probe (35 mm in diameter) (Stable Micro Systems Ltd,
Godalming, UK). A pre- and post-test speed (5.0 mm/s) and test speed
(10.0 mm/s) were employed with a trigger force of 5 g and a com-
pression distance of 25 mm. The pause between the 1st and 2nd com-
pression was 1 s. The textural analysis was conducted on whole fresh
CSB 30 min after steaming and also on whole CSB stored at 4 °C for
24 h. The probe pressed from top of CSB to cause deformation. The
staling rate (%) was calculated by the increase in hardness of CSB after
storage divided by the hardness of the fresh CSB.
2.3.7. Microbiological shelf life of CSB
The microbiological shelf life of CSB was determined by the method
of Hager et al. (2012). Briefly, CSB was sliced transversely from the
centre to obtain a 25 mm slice. The slice was kept in a sealed zip lock
bag with a cut plastic dropper creating a small hole to allow comparable
aerobic conditions. The samples were put at room temperature
(∼20 °C) and inspected daily for mould growth for 5 days. The shelf life
of the CSB was defined as the period before the 1st appearance of
mould.
may facilitate the interactions among amylose-amylose and amylose-
mucilage during the cooling phase of pasting event. The pasted gel was
stored at 4 °C for 24 h. The development of gel texture of wheat flour at
this stage is largely due to the amylose retrogradation. The replacement
by chia seeds greatly decreased the hardness (HD) and adhesiveness
(ADH) of wheat flour gel (Table 1). The much reduced HD and ADH
could be largely attributed to the decreased amounts of starch and
protein available for the gelation. The protein in the chia seeds was
unavailable for gelation. The chia seeds in the form of solid particles
tended to disrupt the gel network. The gel formed from the chia seed
mucilage was apparently not as strong as that of the wheat flour. In-
deed, Capitani et al. (2015) found that the gel of chia seed mucilage was
of shear-thinning nature. The effect of chia seed replacement on the
pasting and gel texture properties of wheat flour largely agreed with
some of the previous studies of starch-hydrocolloid composites
(BeMiller, 2011).
3.2. Effect of chia seed on glycemic response and in vitro starch digestibility
of CSB

2.3.8. Sensory evaluation of CSB
A discriminative sensory testing of the fresh CSB was performed
with 19 panelists (9 females and 10 males). The panelists are of Asian
ethnicity with CSB as a regular part of the diet. Training was provided
at the beginning of the 1st evaluation session. The evaluation ques-
tionnaire of 15-cm line scales and the description of each terms on the
questionnaire are provided in Supplementary Table 2 and
Supplementary Table 3, respectively. The sensory evaluation was con-
ducted independently on 3 different days.
2.4. Data analysis
All the tests were conducted at least in triplicate. The differences
between means of different data were analyzed using the Tukey test
(p < 0.05) on SPSS software (IBM Corporation, New York, USA).
3. Results and discussion
3.1. Effect of chia seed on rheological and gel textural properties of wheat
flour
The replacement by chia seeds increased the peak viscosity (PV),
hot past viscosity (HPV), cool paste viscosity (CPV), and setback (SB) of
wheat flour during pasting event, while having little effect on the
breakdown (BD) (Table 1). The mucilage of chia seeds, which is mostly
concentrated in the first layers of the seed coat, is released upon the
contact with water (Muñoz et al., 2012). Upon hydration, the mucilage,
with a high degree of water binding capacity, develops a high viscosity
(Capitani et al., 2015). The increased SB indicated that the mucilage
The chia seed replacement up to 300 g/kg greatly affected the in
vitro starch digestibility and expected glycemic index (eGI) of CSB
(Table 2). In a dose-dependent manner, chia seed addition decreased
the contents of rapidly digestible starch (RDS) (from 278 g/kg down to
215 g/kg) and slowly digestible starch (SDS) (from 197 g/kg down to
95 g/kg). The resistant starch (RS) content was decreased from 236 g/
kg to 192 g/kg when the chia addition level was at 300 g/kg. The re-
duced RDS, SDS, and RS contents could be largely attributed to the
reduced total starch content of CSB as a result of chia replacement. The
presence of chia seed mucilage may increase the viscosity of the CSB
slurry (as shown in section 3.1 above), reducing the physical associa-
tions and interactions between starch and the hydrolytic enzymes
(Singh, Dartois, & Kaur, 2010). Another possibility is the leaching of
other bioactive components (e.g., polyphenols) from the chia seeds
during CSB making, which remain to be studied. The leached compo-
nents may interact with the starch or the enzymes, contributing to the
reduced starch hydrolysis (Singh et al., 2010).
The percentage of hydrolysed starch at the equilibrium (C∞) was
largely reduced by chia addition, though the kinetic constant (k) was
little affected. The expected glycemic index (eGI) of CSB without chia
addition was found to be 117, which was higher than that of the white
bread (eGI = 95). Jiao et al. (2003) analyzed the GI of a series of staple
cereal foods in humans from Tianjin City of China and found that the
CSB had a high GI of 98.3. The difference of GI between this study and
that of Jiao et al. (2003) may indicate the difference in the experi-
mental methods (e.g., in vitro vs in vivo) and also the CSB production
method (e.g., steaming and fermentation time). Nevertheless, these
studies all indicate that CSB has a high GI (> 70). The addition of chia
seeds at 300 g/kg significantly reduced the eGI of CSB by a percentage

Table 1
Pasting and gel textural properties of wheat flour incorporated with intact chia seeds.
Chia level (g/kg) Pasting Gel texture

PV (Pa·s) HPV (Pa·s) CPV (Pa·s) BD (Pa·s) SB (Pa·s) Hardness (g) Adhesiveness (g·s)

0 5.78 ± 0.04 a 2.38 ± 0.04 a 5.40 ± 0.09 a 3.40 ± 0.01 a 3.02 ± 0.05 a 29.4 ± 2.2 d −254 ± 25 f
25 6.63 ± 0.12 bcd 2.67 ± 0.09 ab 5.98 ± 0.14 bc 3.96 ± 0.03 a 3.32 ± 0.05 a 24.4 ± 1.5 c −262 ± 12 ef
50 6.27 ± 0.13 ab 2.62 ± 0.15 ab 5.86 ± 0.04 b 3.65 ± 0.28 a 3.25 ± 0.11 a 22.8 ± 0.2 c −272 ± 25 e
75 7.05 ± 0.04 cd 2.99 ± 0.19 bc 6.40 ± 0.16 c 4.06 ± 0.16 a 3.41 ± 0.03 a 21.8 ± 2.6 c −297 ± 21 d
100 7.26 ± 0.25 d 3.30 ± 0.08 cd 7.26 ± 0.08 d 3.96 ± 0.17 a 3.96 ± 0.01 b 20.7 ± 1.1 bc −331 ± 55 c
150 7.26 ± 0.20 d 3.41 ± 0.14 cd 7.32 ± 0.30 d 3.85 ± 0.06 a 3.91 ± 0.16 b 17.0 ± 0.7 ab −392 ± 77 b
200 8.05 ± 0.02 e 3.73 ± 0.34 de 7.88 ± 0.11 e 4.32 ± 0.36 a 4.15 ± 0.23 b 15.7 ± 2.5 a −472 ± 83 a
300 7.82 ± 0.19 e 4.01 ± 0.16 e 8.25 ± 0.37 e 3.96 ± 0.02 a 4.24 ± 0.21 b 15.5 ± 1.8 a −463 ± 45 a

Pasting and gel textural properties of wheat flour-intact chia seed blends are presented in this table; PV, peak viscosity; HPV, hot paste viscosity; CPV, cool paste
viscosity; BD, breakdown viscosity; SB, setback; gel texture of pasted flour was measured after 24 h storage at 4 °C; the number of repetitions for the experiment is 3;
the same letter in same column indicates no significant difference between the means at p < 0.05.

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F. Zhu, C. Chan LWT - Food Science and Technology 98 (2018) 77–84

Table 2
Starch nutritional fractions, hydrolysis kinetics, and expected glycemic index of Chinese steamed bread (CSB) incorporated with intact chia seeds.
−1
Chia level (g/kg) RDS (g/kg) SDS (g/kg) RS (g/kg) C∞ k (min ) eGI eGL

0 278 ± 14 c 197 ± 25 c 236 ± 13 b 0.48 ± 0.01 d 0.05 ± 0.01 a 117 ± 2 c 50 ± 2e

50 268 ± 13 bc 161 ± 19 bc 247 ± 11 b 0.41 ± 0.01 c 0.05 ± 0.01 a 106 ± 4 b 43 ± 2d
100 228 ± 20 ab 165 ± 12 bc 247 ± 14 b 0.38 ± 0.01 b 0.05 ± 0.00 a 98 ± 3 ab 38 ± 1c
200 222 ± 18 ab 102 ± 28 ab 249 ± 15 b 0.36 ± 0.02 ab 0.06 ± 0.02 a 95 ± 8 ab 32 ± 1b
300 215 ± 28 a 95 ± 30 a 192 ± 20 a 0.33 ± 0.01 a 0.06 ± 0.01 a 88 ± 3 a 26 ± 1a

In vitro starch digestibility of CSB fortified with intact chia seeds is presented in this table; RDS, rapidly digestible starch; SDS, slowly digestible starch; RS, resistant
starch; the percentages of the starch nutritional fractions are based on the total dry matter of the sample; C∞, percentage of hydrolysed starch at the equilibrium
(120 min) (based on the total dry matter); k, kinetic constant (min −1); HI, hydrolysis index (%); eGI, expected glycemic index; eGL, expected glycemic load; the
number of repetitions for the experiment is 3; the same letter in same column indicates no significant difference between the means at p < 0.05.

of 25% (Table 2), which could be readily attributed to the reduced
starch digestibility as discussed in the last paragraph. The eGI of CSB
with 300 g/kg of chia seeds still had a value of 88, which is considered
high. Processing methods to further reduce the eGI of CSB remain to be
studied. The chia seed addition also significantly lowered the expected
glycemic load (eGL) of CSB from 50 to 26 when the addition level was
at 300 g/kg. The decrease could be readily attributed to the reduced
wheat flour and thus the total starch content as well as the reduced
starch digestibility brought on by chia seed addition as discussed above.
3.3. Effect of chia seed on water activity, volumetric properties, and colour
of CSB
Chia seed replacement of wheat flour reduced specific volume (SV)
of CSB in a dose-dependent manner, though the spread ratio (SR) was
little affected (Table 3). For example, when the chia seed level in-
creased to 300 g/kg, the SV of CSB decreased from 1.878 (control) to
1.371 mL/g. The volume of CSB decreased from 273 mL (control) to
205 mL (with 300 g/kg chia) with increasing level of chia seeds. The
reduced SV could be attributed to replacement of wheat dough by chia
seeds and also to the gluten dilution effect induced by chia seed addi-
tion. The solid particles of chia seeds also disrupted the gluten and
starch network of CSB, reducing the gas holding capacity of the cells.
The mucilage may not be able to form an enough amount of network to
negate the gluten dilution effect. Similarly, a previous study showed
that addition of chia seeds (100 g/kg) decreased the SV of bread by 17%
(Steffolani et al., 2015). The denser structure of CSB with chia seeds
may also contribute to the decreased in vitro glycemic response of CSB
as shown in the above section (3.2). A higher density of CSB reduced
the chance of hydrolytic enzymes to easily penetrate/disperse into
bread matrix to interact with the starch (Fardet, Leenhardt, Lioger,
Scalbert, & Rémésy, 2006).
The moisture content and water activity of CSB were not affected by
the chia seed replacement (Table 3). This may be largely due to the
steaming process fully hydrating the CSB components. The high
moisture content and water activity of CSB largely contributed to the
short shelf life of CSB as discussed in section 3.5 below.
Chia seed replacement much altered the colour of both the crust and
crumb of CSB (Table 4, Fig. 1). Increasing chia concentration to 300 g/
kg decreased the lightness (L*) (from 81 to 68) and b* (from 16.0 to
10.1), while slightly increasing a* of the CSB crust. The total colour
difference (ΔE) tended to increase with the increasing chia replacement
level. Similar to the effect on crust colour, chia replacement at 300 g/kg
somewhat decreased the L* (from 74.7 to 72.8) and b* (from 14.2 to
10.2) of the crumb, while slightly increasing a*. The changes in some of
the colour parameters such as L* and ΔE of crumb appeared to be not in
a linear relationship with the chia replacement level. This may be due
to the uneven distribution of the seeds as the colour-meter only ran-
domly measured particular spots on the samples, rather than the entire
CSB. Chia seeds naturally come in a variety of colours from black-
spotted to white (Muñoz et al., 2013). Therefore, there is potential to
develop a variety of chia fortified CSB varying in colour. It should be
noted that the altered colour may not be an indication of a lower ac-
ceptance by the consumers from the viewpoint of new product devel-
opment, which is described in section 3.6 below. The non-uniform
colour of CSB may not be a negative quality attribute. For example, the
surface of some of the local CSB in China is decorated with sesame.
3.4. Effect of chia seed on textural properties of CSB
Increasing chia seed replacement in CSB increased the hardness
(from 36 N to 112 N) and chewiness (from 27 N to 60 N) of fresh CSB,
while decreasing the cohesiveness (from 0.825 to 0.606) (Table 5). The
large increase in CSB hardness could be mostly attributed to the gluten/
starch dilution effect from chia seed addition. The CSB crumb became
“denser” due to the decreased gas holding capacity of the cells. This is
also reflected by the decreased SV as described in section 3.3 above.
Chewiness is related to the energy needed to masticate a solid food item
until it is ready for swallowing (Iglesias-Puig & Haros, 2013). Addition
of the seeds increased the chewiness of CSB (from 27 N up to 60 N). This
reflects the denser structure of CSB with chia seeds which is chewier
and needs more energy and longer time to bite into small pieces in the
mouth. Similarly, in bread systems, addition of chia seeds increased the
hardness and chewiness of the products (Steffolani et al., 2015). For

Table 3
Specific volume, spread ratio, moisture content, and water activity of Chinese steamed bread (CSB) incorporated with intact chia seeds.
Chia level (g/kg) Specific volume (mL/g) Spread ratio Moisture (g/kg) Water activity

00 1.88 ± 0.05 e 1.754 ± 0.011 ab 416 ± 0a 0.983 ± 0.002 abc

25 1.80 ± 0.02 d 1.799 ± 0.018 ab 412 ± 13 a 0.980 ± 0.002 a
50 1.78 ± 0.03 d 1.821 ± 0.022 b 407 ± 27 a 0.982 ± 0.000 ab
75 1.70 ± 0.03 d 1.715 ± 0.012 ab 414 ± 71 a 0.986 ± 0.002 abc
100 1.67 ± 0.03 c 1.763 ± 0.011 ab 418 ± 71 a 0.984 ± 0.003 abc
150 1.66 ± 0.01 c 1.721 ± 0.009 a 415 ± 71 a 0.987 ± 0.001 c
200 1.53 ± 0.01 b 1.786 ± 0.009 ab 412 ± 71 a 0.987 ± 0.001 bc
300 1.37 ± 0.01 a 1.789 ± 0.012 ab 404 ± 17 a 0.988 ± 0.003 c

The parameters were measured on fresh CSB; the number of repetitions for the experiment is 3; the same letter in same column indicates no significant difference
between the means at p < 0.05.

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Table 4
Colour of crust and crumb of Chinese steamed bread (CSB) incorporated with intact chia seeds.
Chia level (g/kg) Crust Crumb

L* a* b* ΔE L* a* b* ΔE

00 +80.9 ± 0.2 e −0.64 ± 0.04 ab +16.3 ± 0.46 ef +74.7 ± 0.16 d −0.47 ± 0.10 b +14.2 ± 1.0 bc
25 +82.1 ± 0.2 e −0.85 ± 0.14 ab +15.1 ± 0.28 e 1.6 +77.5 ± 0.24 e −0.45 ± 0.12 b +13.7 ± 0.3 bc 3
50 +79.2 ± 0.6 d −0.57 ± 0.18 ab +14.6 ± 0.19 ef 2.5 +75.3 ± 0.83 d −0.26 ± 0.14 c +13.3 ± 1.0 b 1
75 +78.7 ± 0.3 cd −1.00 ± 0.02 a +15.5 ± 0.70 de 2.5 +75.8 ± 0.93 d −0.98 ± 0.02 a +15.3 ± 1.2 c 2
100 +77.6 ± 0.2 c +0.14 ± 0.52 cd +13.9 ± 0.59 cd 4.3 +69.5 ± 0.44 b +0.05 ± 0.02 d +9.6 ± 0.1 a 7
150 +78.1 ± 0.8 cd −0.22 ± 0.29 bc +13.4 ± 0.54 bc 4.1 +66.7 ± 0.98 a +0.22 ± 0.04 d +9.9 ± 0.4 a 9
200 +75.5 ± 0.8 b +0.25 ± 0.26 cd +12.5 ± 0.30 b 6.8 +72.3 ± 0.09 c −0.17 ± 0.04 c +10.3 ± 1.5 a 5
300 +68.1 ± 0.3 a +0.69 ± 0.05 d +10.1 ± 0.06 a 14.4 +72.8 ± 0.28 c +0.11 ± 0.02 d +10.2 ± 1.0 a 4

L*, lightness; a* represents green–red component; b* represents blue–yellow component; ΔE is derived from L*, a*, and b* and represents colour difference between
sample and the control; the number of repetitions for the experiment is 6; the same letter in same column indicates no significant difference between the means at
p < 0.05.

Fig. 1. Photos of fresh CSB fortified with intact chia seeds at different concentrations; A, 0 g/kg; B, 25 g/kg; C, 50 g/kg; D, 75 g/kg; E, 100 g/kg; F, 150 g/kg; G, 200 g/
kg; H, 300 g/kg; CSB, Chinese steamed bread; to show the scale, the height and width of sample A were determined as 53 mm and 94 mm, respectively.

example, a previous study showed that the addition of chia seeds at
100 g/kg increased the hardness of bread by 33% (Steffolani et al.,
2015). Cohesiveness is indicative of the strength of internal bonds
making up the bread crumb (Iglesias-Puig & Haros, 2013). Chia seed
replacement gradually decreased the cohesiveness of CSB in a dose-
dependent manner. This may reflect that the microstructure of bread
was much disrupted due to the gluten/starch matrix dilution by chia.
Overall, the mucilage network from chia seeds appeared to be not
strong enough to counter-balance the gluten dilution effect. It should be
noted there is a great diversity in CSB texture by consumer preference
in China (Zhu, 2014). The altered texture of CSB due to chia seed ad-
dition (e.g., increased hardness) may be a textural advantage to some
consumers as indicated by sensory evaluation described in section 3.6.
The staling of CSB after storage at 4 °C for 24 h was studied by TPA
(Table 5). The storage drastically increased the hardness, chewiness,
while decreasing the cohesiveness of CSB. The staling of CSB is largely
due to the starch retrogradation and water migration/redistribution
during the storage (Zhu, 2016). Chia seed replacement greatly reduced

Table 5
Textural properties of Chinese steamed bread (CSB) incorporated with intact chia seeds before and after storage.
Chia level (g/kg) Fresh CSB CSB stored at 4 °C for 24 h Staling rate (%)

Hardness (N) Chewiness (N) Cohesiveness Hardness (N) Chewiness (N) Cohesiveness

0 36 ± 1 a 27 ± 2a 0.825 ± 0.040 e 235 ± 6a 96 ± 2 a 0.464 ± 0.102 a 553 f

25 43 ± 1 b 32 ± 2 ab 0.837 ± 0.037 e 248 ± 3 ab 116 ± 6 ab 0.556 ± 0.021 a 477 e
50 51 ± 1 c 37 ± 1 bc 0.797 ± 0.008 d 272 ± 1c 120 ± 2 bc 0.542 ± 0.003 a 434 d
75 55 ± 5 cd 40 ± 3c 0.759 ± 0.010 c 255 ± 2b 111 ± 5 ab 0.543 ± 0.023 a 363 c
100 58 ± 1 d 40 ± 1c 0.765 ± 0.010 c 279 ± 5 cd 106 ± 5 ab 0.500 ± 0.026 a 381 c
150 73 ± 1 e 47 ± 2d 0.717 ± 0.029 b 289 ± 2d 126 ± 4 bc 0.534 ± 0.010 a 296 b
200 93 ± 2 f 58 ± 2e 0.698 ± 0.001 b 312 ± 6e 141 ± 5 c 0.546 ± 0.023 a 235 a
300 112 ± 6 g 60 ± 5e 0.606 ± 0.011 a * * * *

*, measurement was beyond measurable range of the texture analyser; the number of repetitions for the experiment is 3; values with the same letter in same column
have no significant difference between the means at p < 0.05.

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F. Zhu, C. Chan LWT - Food Science and Technology 98 (2018) 77–84

the staling rate of CSB from 553% (control) to 235% (300 g/kg chia),
largely in a dose-dependent manner (Table 5). Iglesias-Puig and Haros
(2013) also noted that addition of chia seeds or chia flour reduced the
staling of bread and the re-crystallization of amylopectin component.
The chia mucilage may interact with starch molecules, inhibiting the
retrogradation. The starch-protein interactions, which are considered a
factor of bread staling, may also be reduced by the presence of chia
seeds and the mucilage (Zhu, 2016). The fresh CSB with or without chia
seeds had rather similar water contents and water activity (Table 3).
The mucilage network with a high water holding capacity may slow the
water migration process during CSB staling. Furthermore, the increased
density of CSB fortified with chia seeds may limit the moisture re-dis-
tribution in CSB during the storage. Indeed, it has been well established
that some food hydrocolloids such as polysaccharides can be effective
antistaling agents in breadmaking (Ferrero, 2017).
3.5. Effect of chia seed on microbiological self-life of CSB
The CSB had very high moisture contents and water activity
(Table 3). Therefore, it has a rather limited shelf life of less than 3 days
(Supplementary Table 4). In this study, mould appeared in the control
CSB without chia seeds on the 5th day of storage. Supplementary
Table 4 showed the development of mould on slices of control CSB and
CSB with 20% chia seed addition. Overall, the chia seed replacement at
levels of < 200 g/kg had no significant effect on the microbiological
shelf life of CSB. This is not a surprise since no antimicrobial compo-
nents are found to be associated with the intact chia seeds and the
mucilage. When the replacement level was over 200 g/kg, a decrease of
shelf life to 2 days was recorded for the CSB (data not shown). This may
be attributed to the high water holding capacity of the chia mucilage,
facilitating the microbial growth during storage. Future work (e.g.,
using functional ingredients) on how to improve/maintain the micro-
biological shelf life of CSB with a high level of chia is needed.
3.6. Effect of chia seed on sensory properties of CSB
Chia seed replacement affected different sensory quality attributes
of CSB to different degrees (Table 6). Increasing chia replacement level
drastically decreased the whiteness of both skin and crumb. This agrees
with the results of colour-meter as described in section 3.3. Chia seeds
significantly decreased the smoothness of CSB due to the presence of
the intact seeds on the surface (Fig. 1).
Increasing chia level appeared to decrease the stickiness of CSB.
This may be due to the dilution effect of CSB starch-gluten network
caused by chia which resulted in reduced stickiness of the bolus during
oral processing. Increasing chia level also increased the hardness of
CSB. This agreed with the results of textural profile analysis as de-
scribed in section 3.4 above. The molecular basis for such an increase is
also discussed in that section. The chia replacement had no significant
effect on the cohesiveness and chewiness of CSB revealed by statistical
analysis (p < 0.05). Therefore, these changes, which could be detected
by instrumental analysis (TPA) shown in section 3.4, could not be
sensed by the panellists.
Chia seeds at a higher level (300 g/kg) increased the bitterness of
CSB. This may be due to the increased viscosity of CSB bolus/pieces
caused by the mucilage, and thus the reduced starch hydrolysis rate by
salivary α-amylase and less maltodextrins produced in the mouth. Also,
some components with bitter taste (e.g, polyphenols) may be released
as the seeds were broken down during chewing. In some types of CSB
such as those in southern part of China, sugar is added as an ingredient
for the CSB formulation (Zhu, 2014). Chia seeds at high levels may be as
well incorporated into the formulation of this kind of CSB, as the sugars
and other ingredients may mask the bitterness. Intact chia seeds with
the confinement of the bitterness in the whole seed would be more
suitable for the formulation of northern style CSB in which there is
usually no sugar addition. Chia seeds have a mild nutty taste. Indeed,
increasing chia levels to 300 g/kg and 200 g/kg increased the scores of
the earthy and nutty aroma.
For the overall acceptability which is the most important quality
attribute, CSB with chia seed replacement at all levels had similar
scores as the control (p < 0.05). Therefore, it is feasible to add up to
300 g/kg of chia seeds in CSB formulation for enhanced nutritional
benefits without compromising the eating quality. For consumers,
health-consciousness may even psychologically counter-balance some
of the negative aspects of CSB such as increased bitterness, which re-
main to be studied on a commercial scale.
3.7. Other considerations
Previous studies showed that the functional ingredients added into
CSB formulation may partially get lost due to the processing (e.g.,
steaming and fermentation) (Zhu et al., 2016). For example, the in vitro
antioxidant activities of black tea got partially lost in CSB due to the
processing (Zhu et al., 2016). Especially, chia seeds are rich in un-
saturated lipids and polyphenols which are susceptible to oxidation
under high temperature/moisture processing. Therefore, changes in the
nutrients of the chia seeds in CSB induced by processing remain to be
studied.
There is a trend to employ intact seeds to formulate bakery pro-
ducts. The nutrients of the seeds tend to be less bioavailable due to the
physical steric hindrance. The intactness of cell walls play a role in
reducing the nutrient release (Dhital, Bhattarai, Gorham, & Gidley,

Table 6
Sensory evaluation scores for fresh Chinese steamed bread (CSB) incorporated with intact chia seeds.
Quality parameters Proportion of intact chia seed in CSB

0 g/kg 25 g/kg 50 g/kg 100 g/kg 200 g/kg 300 g/kg

Whiteness Skin 8.0 ± 0.9 d 7.2 ± 1.2 cd 6.5 ± 1.2 c 5.0 ± 1.2 b 4.4 ± 1.4 b 2.0 ± 1.5 a
Crumb 8.1 ± 0.9 e 7.3 ± 1.1 de 6.6 ± 1.1 d 5.2 ± 1.2 c 4.1 ± 1.4 b 2.2 ± 1.6 a
Smoothness 8.5 ± 1.0 d 7.2 ± 1.8 c 6.5 ± 1.2 c 4.4 ± 1.9 b 3.2 ± 1.9 b 1.6 ± 1.6 a
Aroma Earthy 1.7 ± 1.5 a 2.8 ± 2.5 a 3.0 ± 2.0 a 3.1 ± 2.4 a 3.1 ± 2.3 a 3.5 ± 2.7 a
Nutty 1.6 ± 1.6 a 2.2 ± 2.1 a 2.3 ± 1.9 a 2.7 ± 1.9 a 3.5 ± 2.4 a 2.8 ± 2.7 a
Hardness 7.3 ± 1.6 c 5.5 ± 2.2 b 5.5 ± 1.7 b 3.9 ± 1.9 ab 4.2 ± 1.7 ab 2.9 ± 1.8 a
Cohesiveness 3.6 ± 2.7 a 3.3 ± 2.1 a 2.9 ± 2.0 a 3.2 ± 2.6 a 3.2 ± 2.4 a 3.6 ± 3.0 a
Stickiness 4.3 ± 2.6 a 4.0 ± 2.4 a 4.7 ± 2.5 a 3.7 ± 2.1 a 4.2 ± 2.1 a 4.0 ± 2.4 a
Chewiness 5.4 ± 1.7 a 5.3 ± 2.6 a 5.6 ± 2.0 a 5.8 ± 2.1 a 6.0 ± 2.1 a 5.8 ± 2.8 a
Sweetness 1.9 ± 1.5 a 2.8 ± 2.1 a 2.9 ± 2.5 a 1.9 ± 1.8 a 2.0 ± 1.7 a 2.5 ± 2.5 a
Bitterness 0.9 ± 1.5 a 0.9 ± 1.2 a 1.1 ± 1.6 a 1.6 ± 2.3 a 1.1 ± 1.5 a 1.8 ± 2.0 a
Acceptability 6.1 ± 2.0 a 6.0 ± 1.9 a 5.6 ± 1.1 a 5.0 ± 1.2 a 5.2 ± 1.8 a 5.1 ± 1.7 a

The number of repetitions for the experiment is 3; the same letter in same row indicates no significant difference between the means at p < 0.05.

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F. Zhu, C. Chan LWT - Food Science and Technology 98 (2018) 77–84

Table 7
Effect of chia seed flour replacement on physicochemical properties of wheat flour and Chinese steamed bread (CSB).
Chia level (g/kg) Hardness (g) (flour) eGI Specific volume (mL/g) Water activity L* (crust) Hardness (N) (fresh CSB) Hardness (N) (staled CSB)

00 29.4 ± 2.2 e 117 ± 2 e 1.880 ± 0.529 d 0.983 ± 0.002 a +80.9 ± 0.18 g 35.7 ± 0.6 a 234 ± 6b
25 16.8 ± 0.4 d – 1.677 ± 0.091 c 0.983 ± 0.004 a +76.1 ± 0.66 f 49.7 ± 4.2 b 212 ± 2a
50 15.6 ± 0.5 d 105 ± 3 d 1.620 ± 0.072 c 0.981 ± 0.000 a +71.5 ± 1.38 e 53.3 ± 3.8 b 228 ± 2 ab
75 15.4 ± 0.2 d – 1.580 ± 0.010 c 0.980 ± 0.002 a +64.7 ± 0.11 d 63.3 ± 3.2 c 226 ± 2 ab
100 12.9 ± 1.2 c 99 ± 4 c 1.453 ± 0.021 b 0.981 ± 0.001 a +63.6 ± 2.02 d 79.9 ± 1.8 d 262 ± 1c
150 11.4 ± 0.9 bc – 1.433 ± 0.006 b 0.982 ± 0.000 a +60.9 ± 0.21 c 92.9 ± 1.8 e 282 ± 7c
200 9.8 ± 1.2 b 84 ± 4 b 1.357 ± 0.153 ab 0.984 ± 0.002 a +56.7 ± 0.25 b 101.1 ± 0.6 f 328 ± 7d
300 6.3 ± 0.4 a 76 ± 5 a 1.287 ± 0.049 a 0.983 ± 0.001 a +51.9 ± 0.01 a 130.5 ± 1.4 g 341 ± 5d

Hardness (g) (flour), gel hardness of wheat flour; eGI, expected glycaemic index of CSB; specific volume of CSB (mL/g); L* (crust), lightness of the colour of CSB crust;
hardness (N) (fresh), hardness of fresh CSB; hardness (N) (staled), hardness of CSB stored at 4 °C for 24 h; -, not detected; the number of repetitions for the experiment
is 3 expect for the colour (6); values with the same letter in same column have no significant difference between the means at p < 0.05.

2016). The oral mastication may partially break down the intactness of
chia seeds in CSB. The nutrients such as proteins and unsaturated lipids
in the chia seeds may only be partially metabolised, though the ma-
jority of chia may enter the large intestine for fermentation. How the
nutrients of the intact chia seeds and the fermentation may improve
human health remain to be better studied.
Ground chia seed flour may be better utilised in human digestive
tract due to the much reduced particle size. Chia seed flour has been
used in the formulation of bread (Iglesias-Puig & Haros, 2013; Steffolani
et al., 2015). In this study, ground chia seed flour has been incorporated
in CSB formulation as well (Supplementary Fig. 1). Influence of chia
flour replacement up to 300 g/kg on some of the physicochemical
properties of wheat flour and CSB is summarised (Table 7). Overall, it
seemed that the chia flour had similar effects on the flour and CSB
quality as compared to intact chia seeds, though to different extents.
For example, chia flour decreased the gel hardness, specific volume,
lightness of CSB, while increasing the CSB hardness. In particular, the
chia flour decreased the eGI of CSB to a lager extent (e.g., up to 76) than
the chia seeds. This may be due to the interactions of starch/enzymes
with the components of chia flour which are more available for their
physicochemical contacts. The small difference in the nutritional
composition of both chia seed types may also have contributed to the
differences in the extents. However, sensory analysis of the CSB showed
that chia flour addition greatly reduced the overall acceptability
(Supplementary Table 5). The total score of acceptability of CSB for-
tified with ground chia flour was significantly lower than that of CSB
with chia seeds by ∼40% when the replacement level was 300 g/kg. In
particular, the bitterness of the former was much stronger than that of
the latter, possibly due to the much increased exposure of chia poly-
phenols and others in the mouth. The unsaturated lipids, which are
much more exposed in the flour, may be oxidized during the fermen-
tation and steaming of CSB, contributing to the decreased acceptability.
Therefore, methods of masking the bitterness of chia flour are needed in
order to make the chia flour a feasible ingredient for “healthy” CSB
formulation and practical applications.
Currently, the cost of chia seeds is relatively high. For example, in
New Zealand, the price of the intact white chia seed used in this study is
US$ 1.4 per 100 g. It would be expected that CSB with chia seed for-
tification is relatively costly. Large scale production of the seeds in
different areas such as China would be expected to decrease the pro-
duction cost of chia seed fortified CSB.
4. Conclusions
This study is the first report on the effect of chia seed on CSB
quality. Incorporation of chia seeds greatly influenced the physico-
chemical properties of wheat flour and CSB. Chia seed is a promising
ingredient for the formulation of “novel” and “healthy” CSB with ac-
ceptable eating quality and potential in reducing glycemic index and
glycemic load. The results of this study will be of importance to aid the
large number of people with chronic diseases such as diabetes. For
practical and commercial applications, the next step would be to con-
duct sensory studies on consumer panel.
Appendix A. Supplementary data
Supplementary data related to this article can be found at https://
doi.org/10.1016/j.lwt.2018.08.016.
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