Food Chemistry 214 (2017) 242247

Contents lists available at ScienceDirect

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

A method for producing superfine black tea powder with enhanced

infusion and dispersion property
Weihua Xiao, Yang Zhang, Chongxin Fan 1, Lujia Han
College of Engineering, China Agricultural University, Box 191, Beijing 100083, PR China

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

Article history: Tea is rich in healthy components including polyphenols, caffeine, gallic acids, and others. Current tech-
Received 19 February 2016 nology of tea infusion and extraction leads to more than 40% soluble solids wasted in spent leaf. To
Received in revised form 19 June 2016 increase the bioaccessibility of black tea, we report a method of pulverization treatments including
Accepted 12 July 2016
general and superfine grinding to reduce the particle size. In comparison with coarsely ground black
Available online 14 July 2016
tea powders (BTPs), the superfine ground BTP with medium diameter 6.9 lm resulted in significant
higher infusion yield of total polyphenols (TPP), caffeine, and water-soluble carbohydrate (WSC). The
Keywords:
total water-soluble solids (WSS) of superfine BTP infusion increased markedly by twice due to the
Black tea powder
Superfine grinding
accelerated diffusion and enhanced solubility. High correlation between particle size and sedimentation
Particle size ratio suggested improved dispersion stability of superfine BTP. The optimal dispersion of 0.1% superfine
Infusion BTP in water was obtained by combination of homogenization and 0.08% CMC-Na formulation with
Dispersion 27.05% centrifugal sedimentation ratio.
2016 Published by Elsevier Ltd.

1. Introduction
Tea is the most widely consumed beverage in the world after
water (McKay & Blumberg, 2002). Apart from being refreshing,
tea also confers a number of health benefits including preventing
the development of cancer, cardiovascular disease, and other
pathologies (Hodgson et al., 2012; McKay & Blumberg, 2002; Pan
et al., 2013; Tijburg, Mattern, Folts, Weisberger, & Katan, 1997;
Yang & Landau, 2000). The refreshing and health benefits of tea
are primarily related to the water soluble components such as
polyphenols, caffeine, amino acids (de Mejia, Ramirez-Mares, &
Puangpraphant, 2009). Numerous factors are known to affect the
release of active components of tea, such as the variety, growing
environment, manufacturing conditions, and grade (particle size)
of leaf tea particles (Astill, Birch, Dacombe, Humphrey, & Martin,
2001). Among them, the particle size is a key parameter associated
with the diffusion of active components from tea leaves.
Superfine grinding has the ability to reduce the particle size of
food ingredients to within the range of 1 nm100 lm (Zhao
et al., 2009). Compared with traditional mechanical grinding meth-
ods, superfine grinding is reported to increase the extraction and
dispersibility (Park, Imm, & Ku, 2001; Zhao, Yang, Gai & Yang,
Corresponding author.
E-mail address: hanlj@cau.edu.cn (L. Han).
1
Present address: Shenyang Xinxu Trading Co., Ltd, Shenyang, Liaoning 110148,
PR China.
2009; Zhu, Huang, Peng, Qian, & Zhou, 2010). The underlying
mechanism can be integrated from the increase of particle surface
area and the breakdown of the cell walls, both leading to improved
bioavailability and bioactivity in vivo or in vitro (Hu, Chen, & Ni,
2012; Tao et al., 2014). It is reported that superfine grinding of
green tea markedly increased the extraction of total polysaccha-
ride, leading to improved antioxidant activity against OH
(Hu et al., 2012). The dispersibility of green tea powder was
effectively enhanced by decreasing mean particle diameter to less
than 10 lm (Park et al., 2001). Considering that black tea accounts
for about 67% of the worlds tea production (Chang, 2015), we
investigate the effects of grinding on the black tea properties
including the morphology, the chemical components, infusion
property and the dispersion performance.
2. Materials and methods
2.1. Materials
Raw black tea (Qimen black tea) was purchased from Shenzhen
Shenbao Huacheng Company (Shenzhen, China). The moisture con-
tent of the original tea sample was 8.0% (w/w). Four stabilizing
agents, sodium carboxymethyl cellulose (CMC-Na), sodium
alginate, agar, and xanthan gum were purchased from Hefei Siyou
Food Industry and Trade (Hefei, China). The standard compounds
of caffeine were obtained from Sigma Aldrich (C0750). All other

http://dx.doi.org/10.1016/j.foodchem.2016.07.096
0308-8146/ 2016 Published by Elsevier Ltd.
 W. Xiao et al. / Food Chemistry 214 (2017) 242247
reagents (Beijing chemical plant, Beijing, China) were of analysis
grade.
2.2. Grinding method
Raw black tea were milled into coarse black tea powder (BTP)
particles by a Retsch ZM100 centrifugal grinding mill (Germany)
through apertures sized in 1.00, 0.50, 0.25, 0.12 mm at
12,000 rpm. Different particle sizes of coarsely ground BTPs were
obtained, denoted as CG1.00, CG0.50, CG0.25, and CG0.12 accord-
ing to the apertures sizes. CG1.00 was then mixed with zirconium
oxide balls (610 mm in diameter) in a volume ratio of 1:2 and
ground for 8 h to prepare the superfine powder using a vibration
ball mill CJM-SY-B (Qinhuangdao Taiji Ring Nano-Products Co.
Ltd, Hebei, China). During the ball milling, the tank was cooled to
30 C with recycled coolant liquid. The BTP samples were
air-dried at 45 C for 48 h and then stored in a refrigerator in
polyethylene bags for further experiments and measurements.
The moisture contents of the BTPs from coarse to fine were
5.70%, 5.75%, 5.59%, 4.18%, and 6.13% respectively.
2.3. Particle size measurements
The particle size distribution was measured from 0.04 lm to
2000 lm using a LS230 laser diffraction particle size analyzer
(Beckman Coulter, Brea, CA, USA) at room temperature with water
as solvent.
Particle size distribution is characterized by the median diame-
ter (D50) and by the span factor [(D90
D10)/D50], where D10, D50,
or D90 values represent 10%, 50% or 90% cumulative percentiles
of particles (from 0 to 100%) undersize particle size distribution
(Mingard et al., 2009). Two measurements were carried out for
each sample.
2.4. Cell wall breakage ratios
Generally, a diameter of 10  20 lm may be regarded as a
typical cell size for plants (Metzler, 2003). When mean particle size
D50 > 10 lm, cell wall breakage ratio U was calculated using the
following equation (Liu, 2007):
U 1
1
10=D50 3 1 the sediment ratio based on the weight of BTPs. The supernatant
When D50 < 10 lm, U = 100%.
2.5. Scanning electron microscope (SEM)
BTPs samples were spread on a conductive adhesive carbon
tape, pasted on a sample stub. The morphological characteristics
of different sized BTPs were investigated by a Hitachi S3400 model
SEM (Hitachi, Japan) at 15 kV.
2.6. BTP infusion preparation and component analysis
To monitor the dissolution of different components during the
BTPs infusion, 0.5 g of BTP was put into a glass flask with 75 ml
of water (25 C) and was mixed thoroughly. The flask was placed
in a water bath at 25 C without agitation.
The mixture was filtered through a 0.45 lm films after 5, 10, 15,
30, 45, 60, and 90 min of leaching for the analysis of water-soluble
carbohydrate (WSC), total polyphenols (TPP), and caffeine. Mean-
while, the parallel sample solutions were filtered to collect the
residue for water-soluble solids (WSS) determination.
WSS were determined by oven drying of the filtered residue at
105 C to constant weight according to the Chinese National
Standard GB/T 8305 (2013).
243
WSC was determined by the anthronesulfuric acid method
using glucose as a standard, as described by McDonald and
Henderson (1964). A Shimadzu UVvis 2550 spectrometer
(Shimadzu, Kyoto, Japan) was used for the UVvis spectroscopic
analysis. The detector wavelength for water-soluble carbohydrate
was 620 nm.
TPP were determined according to the Folin-Ciocalteu method
(Singleton & Rossi, 1965) with gallic acid as calibrant. The detector
wavelength was set to 765 nm.
The caffeine content of the BTP infusions was determined
according to ISO 14502-2 (2005). A Hitachi HPLC system (L-7000,
Hitachi, Japan) was used to analyze the solution using binary
gradient elution at a flow rate of 1 mL/min. The mobile phase A
was 9% (v/v) acetonitrile, 2% (v/v) acetic acid with 20 lg/mL EDTA.
The mobile phase B was 80% (v/v) acetonitrile, 2% (v/v) acetic acid
with 20 lg/mL EDTA). An Agilent ZORBAX SB-C18 column
(4.6  250 mm, 5 lm) was used for separation and the detector
wavelength was set to 278 nm.
All the results were expressed as a mass percentage of BTPs on
dry basis.
2.7. Dispersion methods and sediment ratios measurement
BTPs were dispersed in deionized water at tea-water ratio of
1:1000 and mixed with agitation for 10 min at room temperature
to form 0.1% tea-water mixture. To improve the dispersity of
superfine BTP, Four stabilizers were investigated including sodium
carboxymethyl cellulose (CMC-Na), sodium alginate, agar, and
xanthan gum. The stabilizers were added respectively to the
tea-water mixture to reach concentrations of 0.08
0.24% (w/w).
To investigate the effect of homogenization on the dispersion
of superfine BTP, an ATS GL-20G-C high-pressure homogenizer
(ATS Engineering, Canada) was utilized at 500 bars for the 0.1%
tea-water mixture with or without stabilizer.
The mixtures of different treatments were then collected for the
measurement of particle sizes and sediment ratios.
The BTPs suspended mixture prepared with above dispersion
procedure were allowed to settle by standing 2 h to determine
gravitational sediment ratio or centrifuge at 4000 rpm for 10 min
to determine the centrifugal sediment ratio. The supernatant was
removed and the sediment was dried and weighed to determine
after centrifugation was also used for particle size measurement.
2.8. Statistical analysis
All experiments were conducted in duplicates. Results were
reported as their replicate means standard deviation (SD). The
differences in mean were calculated using the Duncans multiple-
range tests with 95% confidence limit (p < 0.05) by SPSS 10.0 software.
The correlation analysis was done by Microsoft Excel 2010.
3. Results and discussion
3.1. Particle size distribution of BTPs and cell wall breakage ratio
Table 1 and Fig. 1 supplementary showed the particle size dis-
tributions of ground BTPs. The particle size of the superfine BTP
was in the range of 2.4
13.1 lm with a median particle size of
6.9 lm. The particle size distributions of coarsely ground powder
fractions of CG1.00, CG0.50, CG0.25, and CG0.12 were much wider,
ranging from 9.4 to 1211.0 lm and their median particle sizes
were 580.7, 306.5, 242.5, and 154.5 lm, respectively. In addition,
the superfine BTP had a significantly narrower particle size
distribution (Span = 1.6) than those of coarsely ground BTPs
244 W. Xiao et al. / Food Chemistry 214 (2017) 242247

Table 1 threaded conduit structures could be clearly observed in CG1.00.

Particle size distributions and cell wall breakage ratios of BTPs. The leaf veins were mostly separated from mesophyll in CG0.50,
Sample CG1.00 CG0.50 CG0.25 CG0.12 Superfine CG0.25, and CG0.12, with the length of ca. 200 lm. After superfine
D50 (lm) 580.7 306.5 242.5 154.5 6.9 grinding, the particle size was significantly reduced and the
D90 (lm) 1211.0 611.2 542.4 399.4 13.1 uniformity was greatly improved as the leaf vein was also pulver-
D10 (lm) 145.3 48.9 22.3 9.4 2.4 ized. The SEM results were highly consistent with the particle size
Span 1.8 1.8 2.1 2.5 1.6 analysis data. Partial aggregation was observed in the superfine
Cell wall breakage 3.0 5.9 11.5 22.1 100.0
ratio (%)
BTP due to high specific surface area and electrostatic interaction
(Jha, Jha, Chaudhury, Rana, & Guha, 2014).

(Span = 1.8
2.5). That the average particle size of superfine BTP
was less than the cell diameter indicates that the cell walls of
superfine BTP were mostly broken.
The cell wall breakage ratio was calculated using Eq. (1) and the
results were showed in Table 1. All the tea leaf cells were ruptured
during the superfine grinding while only 3.022.1% of the cell wall
were broken by general grinding.
3.2. Morphology of BTPs
SEM images of 200 times magnification were obtained for the
five sizes of BTPs and given in Fig. 1. Vascular bundles, and
3.3. Infusion property of BTPs
The contents of TPP, caffeine, WSC, and WSS extracted as a func-
tion of infusion time were showed in Fig. 2. Generally, the typical
infusion curves of tea components displayed high diffusion
efficiency at the initial stage and gradually levelled off with
increasing time to 90 min. Superfine BTP reached the maximum
yield at 5 min while it took over 15 min for the coarsely ground
BTPs to obtain the infusion equilibrium. Superfine BTP yielded
significantly higher levels of TPP, caffeine, WSC, and WSS than
coarsely ground BTPs as shown in Fig. 2.
The TPP yields of BTPs increased by reducing particle size. TPP
in the superfine BTP infusion at 5 min was up to 7.0%, while that

Fig. 1. SEM images for various particle sizes of BTPs in 200 magnification. (A) CG1.00; (B) CG0.50; (C) CG0.25; (D) CG0.12; (E) superfine powder.
 W. Xiao et al. / Food Chemistry 214 (2017) 242247 245

10 CG1.00 CG0.50 CG0.25 A 12 CG1.00 CG0.50 CG0.25 C

CG0.12 Superfine CG0.12 Superfine

8 10

8
6

WSC (%)
TPP (%)

6
4
4

2
2

0 0
0 15 30 45 60 75 90 0 15 30 45 60 75 90
Time (min) Time (min)

4 CG1.00 CG0.50 CG0.25 B 60 CG1.00 CG0.50 CG0.25 D

CG0.12 Superfine CG0.12 Superfine
50
3
40
Caffeine (%)

WSS (%)
2 30

20
1
10

0 0
0 15 30 45 60 75 90 0 15 30 45 60 75 90
Time (min) Time (min)

Fig. 2. Infusions of different components from BTPs. (A) TPP; (B) caffeine; (C) WSC; (D) WSS.

of CG1.00 at the same time only reached 1.8%. Meanwhile, yields of
the equilibrium were also increased with decreasing particle size.
Similar trends were observed in the caffeine yields. Its commonly
accepted that smaller particles of high specific surface show fast
infusion and reduce the time needed for solute diffusion through
the particle matrix (Smith, 2003). The breakage of cell wall by
superfine grinding also greatly reduces the mass transfer resis-
tance and enhances the diffusion rate of chemical components
(Li, Wei, Liu & Yin, 2007).
For coarsely ground BTPs, the infusion rate of WSC and the
extraction yield at equilibrium both increased significantly with
decreasing particle size. Similar results were observed for the
extraction of polysaccharide from fine Polystictus versicolor powder
(Yuan & Chen, 2005). Research on the effect of superfine grinding
on the physical properties of oat bran soluble dietary fiber
indicated that the solubility of the oat bran soluble dietary fiber
was improved by superfine grinding from 5.9% to 16.6% (Zhang,
Liang, Pei, Gao, & Zhang, 2009). Therefore, the significant increase
in WSC yields of superfine BTP can be contributed to not only
the accelerated mass transfer, but also the depolymerization of
structural polysaccharides by superfine grinding.
The maximum extraction yield of superfine BTP is 47.9% at
90 min which is almost twice of those of coarsely ground BTPs.
Our previous study on wheat straw showed that superfine grinding
changed the microstructure of wheat straw by reducing the
crystallite size and crystallinity of cellulose (Yang, Ji, Xiao, & Han,
2014). Protein solubility of ginger powder was also reported to
increase with decreasing particle size from 300 to 8.34 lm (Zhao
et al., 2009). We can deduce that superfine grinding has modified
the physicochemical properties of macromolecules of BTP and
improved their solubility in water. The high ratio of WSS from
superfine BTP can be partly due to the conversion of insoluble
macromolecules e.g. cellulose and hemicellulose into soluble
fractions.
3.4. Sedimentation performance of BTPs
Particle size is known to influence the sedimentation behavior
and cloudiness of beverages (Neidhart et al., 2002). The correlation
between particle size and sedimentation has been used to deter-
mine the particle size distribution of finely divided materials such
as hydrated lime (Bishop, 1934). Table 2 showed the gravitational

Table 2
Gravitational and centrifugal sedimentation rates of BTPs.

Sample CG1.00 CG0.50 CG0.25 CG0.12 Superfine

Gravitational sedimentation rate (%) 72.92 2.02b 70.83 4.16b 69.79 2.11b 65.62 2.24b 48.96 3.01a
Centrifugal sedimentation rate (%) 77.45 2.74b 73.78 2.35ab 72.92 5.50ab 71.18 2.82a 70.03 3.39a

Data given as percentage of dry mass (w/w), mean standard deviation.

Entries in columns marked with different lowercase letters are significantly different by t test (p < 0.05).
246 W. Xiao et al. / Food Chemistry 214 (2017) 242247

and centrifugal sedimentation ratio of BTPs with different particle 100 10.0
Centrifugal sedimentation ratio
sizes. The gravitational sedimentation ratios decreased sharply 90 9.0
from 72.92% to 48.96% with particle size reducing from CG1.00 to D50
80 8.0
superfine. After standing for 2 h, the aqueous solutions of coarsely
ground BTPs were totally transparent while that of the superfine 70 7.0
BTP was cloudy, which indicated a large amount of BTP particles 60 6.0
suspended in water. A high correlation (r2 = 0.98) between

m
%
50 5.0
gravitational sedimentation ratio and WSS (at 90 min) was
40 4.0
observed. There is a slight decrease in the centrifugal sedimenta-
tion ratio as particle size reduces. The above results suggested 30 3.0
that BTPs of smaller particles could be dispersed in water with 20 2.0
higher stability than those of larger particles. There is a high 10 1.0
correlation (r2 = 0.97) between particle size and centrifugal
0 0.0
sedimentation ratio. CK S H S+H

3.5. Dispersion optimization of superfine BTP
To further improve the dispersion stability of superfine BTP,
several downstream processes were investigated including
addition of stabilizers and homogenization.
Hydrocolloids or gums are widely used as stabilizers in the food
industry for their abilities to increase solution viscosity, retard the
precipitation of dispersed solid particles, and prevent aggregation
of dispersed particles (Valdez, 2012). Four commonly used stabiliz-
ers are CMC
Na, sodium alginate, agar, and xanthan gum. Their
effects on the centrifugation sedimentation ratio of superfine BTP
are shown in Fig. 3. The centrifugal sedimentation ratio decreased
significantly with increased concentration of stabilizers. Adding
low concentration of xanthan gum and CMC
Na (0.08%) signifi-
cantly reduced the centrifugal sedimentation ratio of superfine
BTP solution. The unfavorable aspect with xanthan gum is the high
viscosity at low concentration, which could influence consumers
preference. To maintain an acceptable viscosity, CMC
Na is a more
ideal stabilizer. It reduced the sedimentation of superfine BTP from
70.03% to 58.71% at low concentration of 0.08%.
Homogenization is another method that is widely used in food
production to increase suspension stability by reducing the particle
size (Song, Zhou, Fu, Chen, & Wu, 2013). The effects of homogeniza-
tion and stabilizer are shown in Fig. 4. It is clear that both homog-
enization and CMC
Na (0.08%) formula reduced the centrifugal
sedimentation ratio of superfine BTP, respectively. CMC
Na had
no obvious effect on the particle size distribution of BTP
suspension, while high-pressure homogenization effectively
Fig. 4. Centrifugal sedimentation ratio and median particle size of superfine BTP
with different treatments. CK, 0.1% superfine BTP suspension; S, 0.1% superfine BTP
suspension with 0.08% CMC
Na; H, 0.1% superfine BTP suspension with 500 bar
homogenization; S+H, 0.1% superfine BTP suspension with 0.08% CMC
Na formula
and 500 bar homogenization.
reduced the particle size of the suspension from 6.9 to 3.5 lm.
The high shear and pressure during homogenization could
potentially crush the BTP particle aggregation. Neidhart et al.
reported that the stabilizing effect of pectin can be enhanced by
homogenization treatments for fruit juices and drinks made from
tropical fruits (Neidhart et al., 2002). The combination of homoge-
nization and stabilizer was investigated by homogenizing the 0.1%
superfine BTP solution with 0.08% CMC-Na. The sedimentation
ratio further decreased to 27.05% and the median particle size
was reduced to 1.3 lm with better uniformity. The increased
viscosity of the outer phase increased the impact between the
aqueous and the solid phase, leading to the decrease in BTP particle
size (Gardouh, Ghorab, & Abdel-Rahman, 2012). The synergistic
effect of the stabilizer and homogenization treatment was
observed in our study.
The particle size of superfine BTP in the supernatant after cen-
trifugation was also measured as shown in Fig. 2 supplementary.
They were significantly less than those in the BTP suspension
before centrifugation. The particle sizes in the supernatant of
centrifuged BTP suspension after homogenization with and
without the stabilizer were very small, at 0.2 and 0.1 lm respec-
tively. This indicates the BTP particle size is at nanoscale in the
stable BTP suspension.

80
Centrifugal Sedimentation Rate (%)

70

60 Xanthan gum

50
CMCNa
40

30 Sodium alginate

20
Agar
10

0
0.08 0.12 0.16 0.20 0.24

Concentration (%)

Fig. 3. Effects of different stabilizers on centrifugal sedimentation ratio of superfine BTP.

 W. Xiao et al. / Food Chemistry 214 (2017) 242247
4. Conclusions
The superfine BTP showed markedly fast infusion rates and high
infusion yields for different water soluble components. This is due
to the cell wall breakage in microparticulation and the modifica-
tion of macromolecules into soluble contents. The superfine BTP
also showed a preferable dispersion stability as compared with
coarsely ground BTPs. Homogenization and addition of stabilizers
have further improved the dispersion properties of superfine BTP.
All these improvements result in higher utilization efficiency and
better sensory in black tea consumption.
Acknowledgments
This study was supported by the National Natural Science
Foundation of China (No. 31271611) and the Research Fund
for the Doctoral Program of Higher Education of China (No.
20130008130004).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2016.
07.096.
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