Food Chemistry 178 (2015) 186–194

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

Effect of extrusion on phytochemical profiles in milled fractions

of black rice
Huihui Ti, Ruifen Zhang, Mingwei Zhang ⇑, Zhencheng Wei, Jianwei Chi, Yuanyuan Deng, Yan Zhang
Sericultural and Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences, Key Laboratory of Functional Foods, Ministry of Agriculture, Guangdong Key Laboratory
of Agricultural Products Processing, Guangzhou 510610, PR China

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

Article history: The phytochemical profile and antioxidant activities of unprocessed and extruded milled fractions of
Received 21 May 2014 black rice were investigated. Extrusion increased the free phenolics, anthocyanins and oxygen radical
Received in revised form 14 November 2014 absorbance capacity (ORAC) and decreased the bound forms. The total phenolics, anthocyanins and ORAC
Accepted 19 January 2015
increased by 12.6%, 5.4% and 19.7%, respectively, in bran. Extrusion decreased both free and bound
Available online 24 January 2015
phenolics and anthocyanins while ORAC values decreased by 46.5%, 88.4% and 33.1%, respectively, in pol-
ished rice and by 71.2%, 87.9% and 14.7%, respectively, in brown rice. A total of seven phenolics, gallic,
Keywords:
chlorogenic, vanillic, caffeic, syringic, p-coumaric and ferulic acids, were detected in both forms. Cyanidin
Black rice
Extrusion
3-glucoside (Cy-3-G), cyanidin 3-rutinoside and peonidin 3-glucoside were also detected with Cy-3-G
Phenolic content found in the highest amounts in unprocessed and extruded rice bran. These results provide the basis
Anthocyanin content for the development of different milled fractions of extruded black rice with balanced nutritional charac-
Antioxidant activity teristics for today’s functional food markets.
Ó 2015 Published by Elsevier Ltd.

1. Introduction which have been shown to have beneficial effects in preventing

chronic diseases. As consumers’ awareness of health increases,
Epidemiological studies have consistently shown that the new processed foods containing black rice materials (rice bran,
regular consumption of foods high in whole grains may contribute polished rice and brown rice of black rice) require the presence
to maintaining good health and possibly reduce the risk of chronic of bioactive ingredients to satisfy the demands of health-conscious
diseases such as cardiovascular disease, type II diabetes, obesity consumers.
and cancer (Okarter & Liu, 2010). Whole grain phytochemicals The extrusion of whole grains is becoming more popular in the
are assumed to be the key source of the health benefits of whole food and pharmaceutical industries. Extrusion is a technology that
grains (Wang, Chen, Xie, Ju, & Liu, 2013). affects the microstructure, chemical characteristics, and macro-
Whole-grain phytochemicals consist of carotenoids (lutein, zea- scopic shape of a product. It is a mechanical process that exposes
xanthin, cryptoxanthin, and carotene), phenolics and vitamin E. the extruded material to high temperatures, shear forces and pres-
Phenolics include phenolic acids (p-coumaric, caffeic, ferulic, vanil- sures over a short period of time. The temperatures encountered by
lic, and syringic acids) as well as flavonoids (flavonols, flavonones, the food material in the barrel of the extruder are high enough to
catechins, and anthocyanins) (Goufo & Trindade, 2014; Liu, 2007). gelatinize starch, denature protein and form complexes between
Whole-grain phenolics possess potent antioxidant activities and starch, lipids and proteins (Wolfe & Liu, 2003). In particular, phen-
are able to scavenge free radicals that may increase oxidative stress olics may undergo various changes, thus altering the antioxidant
and potentially damage large biological molecules such as lipids, activity of the products. Many studies have reviewed and docu-
proteins and DNA. Black rice is a kind of typical whole grain, and mented the effects of extrusion on the phenolic contents and anti-
is considered a good source of fiber, minerals and phytochemicals oxidant activity of different foods included thermally processed
as well as basic nutrients (Zhang, Zhang, Zhang, & Liu, 2010). Tra- legumes (Han & Baik, 2008; Xu & Chang, 2008) and extruded barely
ditionally, black foods, particularly black rice, have long been (Sharma, Gujral, & Singh, 2012). It is also well documented that the
favored by Chinese people. The popularity of black rice is partly total phenolic content and antioxidant activity of table beets, green
due to its particular phytochemicals, especially anthocyanins, beans and corn were affected by thermal processing (Jiratanan &
Liu, 2004; Parra, Saldivar, & Liu, 2007).
⇑ Corresponding author. Tel.: +86 2087237865; fax: +86 2087236354. Beneficial phytochemicals in grain and processed grain samples
E-mail address: [email protected] (M. Zhang). are distributed in rice in the free, soluble-conjugated and bound

http://dx.doi.org/10.1016/j.foodchem.2015.01.087
0308-8146/Ó 2015 Published by Elsevier Ltd.
 H. Ti et al. / Food Chemistry 178 (2015) 186–194 187

forms. Most of these free and bound compounds are found in 2.3. Extrusion
different parts of the grain, particularly in distinct fractions
obtained from milling the grains (Onyeneho & Hettiarachehy, Extrusion was performed using a Werner and Pfleiderer Con-
1992). A previous study investigated the phenolics and anthocya- tinua 37 co-rotating twin-screw extruder (Stuttgart, Germany).
nin phytochemicals and their antioxidant activity in different The screw diameter, (L/D) ratio and die diameter were 37 mm,
milled fractions (bran, polished rice and brown rice) of black rice 27/1 and 6 mm, respectively. The feed rate (25 kg/h) and screw
(Kong & Lee, 2010). It should be noted that the extraction proce- speed (200 rpm) were kept constant. The extrusion was carried
dure used in this study may have led to an underestimation of out at 120 °C with the temperature of different barrel zones set
the total phenolics and antioxidant activity because the bound at 60, 100 and 120 °C. The feed moisture was conditioned to 12–
fraction was not included in the analysis (Butsat & Siriamornpun, 17%. The extrudates were cooled to room temperature, packed in
2010). Thus, a better understanding of how thermal processing polyethylene bags and later milled to flour using a grinder (Sujata,
influences the stability of phenolics in black rice and their distribu- India) to a particle size <250 lm and stored at 20 °C until further
tion in the extrudate is required. analysis.
The objectives of the present investigation were to determine
the content of free and bound phytochemicals (phenolics and 2.4. Extraction of free phenolic compounds
anthocyanins) and their antioxidant activities and to analyze the
composition and content of individual phytochemicals in both free Free phenolic compounds in the raw or extruded flours were
and bound forms in different milled fractions (bran, polished rice extracted using the method of Sun, Chu, Wu, and Liu (2002). For
and brown rice) of black rice before and after extrusion. each sample, a total of 0.5 g rice bran and 2 g polished/brown rice
were blended with 50 mL of chilled acidified methanol (95% meth-
anol and 1 M HCl 85:15, v/v). The extract was homogenized using
an XHF-D homogenizer at 10,000 rpm for 5 min in an ice bath
2. Materials and methods
(Ningbo Xin-zhi-Bio Technology Co. Ltd., Ningbo, China) and cen-
trifuged at 2500g for an additional 10 min. The extraction was
2.1. Chemicals and reagents
repeated. The supernatants were removed, pooled and concen-
trated by evaporation at 45 °C. The free phenolics were then made
Analytical grade methanol (MeOH), ethanol (EtOH), hexanes,
up to a final volume of 10 mL with chilled acidified methanol and
ethyl acetate, hydrochloric acid (HCl), acetic acid (HAc), potassium
stored at 20 °C until further analysis.
chloride (KCl), sodium acetate (NaAc), sodium carbonate (Na2CO3),
sodium hydroxide (NaOH), potassium phosphate monobasic (KH2-
2.5. Extraction of bound phenolic compounds
PO4) and potassium phosphate dibasic (K2HPO4) were purchased
from Mallinckrodt Chemicals (Phillipsburg, NJ, USA). 20 ,70 -Dichloro-
The bound phenolic compounds in the different milled fractions
fluorescin diacetate (DCFH-DA), fluorescein disodium salt, apige-
of black rice and their extrudates were extracted according to the
nin, sodium borohydride (NaBH4, reagent grade), chloranil
methods of Naczk and Shahidi (1989) and Sun et al. (2002). The
(analytical grade), vanillin (analytical grade), and catechin hydrate
residue from the free extraction was then digested with 40 mL of
were purchased from Sigma–Aldrich Inc. (St. Louis, MO, USA). Tet-
2 M NaOH at room temperature. The oxygen was removed under
rahydrofuran (THF, analytical grade) and aluminum chloride (AlCl3-
nitrogen gas and the sample was shaken for 1 h. The mixture
6H2O, analytical grade) were purchased from Fisher Scientific (Fair
was extracted using hexanes to remove lipids and then neutralized
Lawn, NJ, USA). Folin–Ciocalteu reagent, 6-hydroxy-2,5,7,8-tetram-
with concentrated hydrochloric acid. The remaining extract was
ethylchroman-2-carboxylic acid (Trolox), trifluoroacetic acid (TFA,
then extracted 5 times with ethyl acetate. The combined superna-
chromatographic grade) and acetonitrile (chromatographic grade)
tants were evaporated under vacuum at 45 °C to dryness. The
were purchased from Sigma. Gallic acid was purchased from ICN
bound phenolics were reconstituted with distilled water to a final
Biomedicals Inc. (Aurora, OH, USA). 2,20 -Azobis (2-amidinopro-
volume of 10 mL and then stored at 20 °C until further analysis.
pane) dihydrochloride (ABAP) was purchased from Wako Chemi-
cals USA Inc. (Richmond, VA, USA). Cyanidin 3-glucoside (Cy-3-G),
2.6. Determination of total phenolic content
cyanidin 3-rutinoside (Cy-3-R), and peonidin 3-glucoside (Pe-3-G)
were purchased from Polyphenols Laboratories (Sandens, Norway).
The total phenolic content of each extract was determined using
the Folin–Ciocalteu (FC) colorimetric method described by
Dewanto, Wu, Adom, and Liu (2002). Briefly, 0.5 mL of distilled
2.2. Grain samples and sample preparation water and 0.125 mL of the above extract were added to a test tube.
FC reagent (0.125 mL) was added to the mixture and it was allowed
Black rice samples (Oryza sativa var. Heiyounian) were to react for 6 min. Then, 1.25 mL of 7% aqueous sodium carbonate
obtained from the Experimental Station of the Rice Research solution and distilled water were added to make up the total vol-
Institute of Guangdong Academy of Agricultural Sciences in ume to 3 mL. The mixture was incubated for 90 min at room tem-
2012. They were sown in late March and harvested in mid-July. perature. The absorbance was read at 760 nm using a Shimadzu
The rice grains were then air-dried to a moisture content of UV-1800 spectrometer (Shimadzu Inc., Kyoto, Japan). Free, bound
approximately 13% and stored at room temperature for 3 months. and total phenolic contents were measured by comparison to a
The rice samples were milled to separate the husk from the standard curve of gallic acid solutions and were expressed as mg
brown rice. The husk was not included in the analysis. The brown gallic acid equivalents (GAE) per 100 g dry weight (DW) of sample.
rice was then polished using a rice milling machine (Satake Co.,
Hiroshima, Japan) to obtain approximately 10% (w/w) rice bran 2.7. Determination of total anthocyanin content
(bran/embryo) and approximately 90% (w/w) polished rice. The
bran, polished rice and brown rice samples were sieved by pass- The total anthocyanin content of each extract was determined
ing through a 60-mesh sieve using a Cyclone Sample Mill (UDY using a spectrophotometric pH differential protocol adapted from
Corporation, Fort Collins, CO, USA) and stored at 40 °C until fur- that of Wolfe and Liu (2003). Briefly, the above extracts were
ther analysis. mixed thoroughly with 0.025 M potassium chloride (pH 1) buffer.
188 H. Ti et al. / Food Chemistry 178 (2015) 186–194

The absorbance of the mixture was then measured at 515 and Pe-3-G in the spiked samples were 95.8%, 97.4% and 97.3%,
700 nm against a distilled water blank. Similarly, the above respectively.
extracts were dissolved with sodium acetate buffer (pH 4.5) and
the absorbance of these solutions was measured at the same wave- 2.10. Determination of antioxidant activity
lengths. The anthocyanin content was calculated by:
The antioxidant activity was determined using the oxygen radi-
Total anthocyanins ðmg=100 g sample DWÞ cal absorbance capacity (ORAC) assay according to a modified
¼ A  MW  DF  1000=ðe  CÞ method from our laboratory (Zhang et al., 2010). Briefly, the above
extract dilutions were prepared daily in 75 mM phosphate buffer
where e is the molar absorptivity of Cy-3-G = 26,900; C is the concen- (pH 7.4). The assay was performed in black-walled 96-well plates
tration of the buffer in mg/mL; A is absorbance = (A515  A700)pH 1.0 (Corning Scientific, Corning, NY, USA). The outside wells of the plate
 (A515  A700)pH 4.5; MW is the molecular weight of Cy-3- were not used to limit well to well variation because of evaporation.
G = 449.2; and DF is the dilution factor. The anthocyanin content Each well contained 20 lL extract or 20 lL Trolox standard
was expressed as the mean (mg Cy-3-G equivalents per 100 g sam- (range = 6.25–50 lm) and 200 lL fluorescein (final concentra-
ple DW) ± SD for triplicate samples. tion = 0.96 lm). The reaction mixture was incubated at 37 °C for
20 min. Then, 20 lL of 119 mM ABAP solution was added using a
2.8. Determination of individual phenolic acids multichannel pipette. The fluorescence intensity was measured
from the top using a Fluoroskan Ascent FL plate-reader (Thermo
Gallic, chlorogenic, vanillic, caffeic, syringic, p-coumaric and Labsystems, Franklin, MA, USA) at an excitation wavelength of
ferulic acids in the sample extracts were quantified using an Agilent 485 nm and emission wavelength of 538 nm for 35 cycles every
1200 HPLC system (Waldbronn, Germany) equipped with an auto- 4.5 min. ORAC was expressed as lmol Trolox equivalents per gram
sampler and Agilent 1200 series VWD detector, using a DW.
250  4.6 mm, 5-lm Agilent Zorbox SB-C18 column (Palo Alto,
CA, USA). Briefly, the gradient elution was conducted with 0.4% 2.11. Statistical analysis
aqueous solution acetic acid (solution A) and acetonitrile (solution
B), programmed as follows: 0–40 min, solution B 5–25%; 40– Data were reported as mean ± SD for triplicate determinations
45 min, solution B 25–35%; 45–50 min, solution B 35–50% at a con- of each sample. ANOVA and the Student–Newman–Keuls (SNK) q
stant flow rate of 1.0 mL/min and a 20 lL sample was injected for all post hoc test were performed. Statistical significance was defined
analyses. The total HPLC run time was 50 min. The UV detection at the level of p < 0.05. All of the statistical analyses were
was set at a wavelength of 280 nm. Samples were filtered through performed using SPSS statistical package version 13.0 (SPSS Inc.
a 0.25-lm membrane filter (Millipore, Billerica, MA, USA) before Chicago, IL, USA).
analysis. Individual phenolic acids were identified on the basis of
retention time and the chromatography of authentic standards. 3. Results
The percentage recovery for these phenolic acids was 94–99%.
3.1. Effect of extrusion on total phenolic content
2.9. Determination of anthocyanin composition
The effects of extrusion on the free, bound and total phenolic
Cy-3-G, Cy-3-R, and Pe-3-G in the sample extracts were quanti- content and the percentage contribution of each fraction to the
fied on a 2695 HPLC system (Waters Corp., Milford, MA, USA) total phenolic content in different milled fractions of unprocessed
equipped with a model 996 photo-diode array detector and model and extruded black rice flours are presented in Table 1.
717 plus autosampler (Waters Corp.), using a C18 column The use of extrusion increased the free phenolic content of rice
(150  3 mm, 3 lm, Phenomenex, Torrance, CA, USA). Briefly, the bran by 17.0% (p < 0.05) and reduced the bound phenolic content
gradient elution was conducted with 4.5% formic acid (solution by 15% (p < 0.05), but overall, the total phenolic content increased
A) and methanol (solution B). The following gradient was used: by 12.6% (p < 0.05). However, there was a significant reduction in
solvent B, 0 min, 10%; 30 min, 25%; 34 min, 33%; 42 min, 90%; the phenolic content of the polished and brown rice samples. The
and 45–50 min, 10%, at a constant flow rate of 0.4 mL/min. A sam- free, bound and total phenolic content decreased (p < 0.05) by
ple volume of 10 lL was injected for all analyses. The total HPLC 53.4%, 25.0% and 46.5%, respectively, in polished rice and by
run time was 50 min. The UV detection was set at a wavelength 79.5%, 24.5% and 71.2%, respectively, in brown rice.
of 520 nm. Samples were filtered through a 0.25-lm membrane fil- Extrusion treatment significantly influenced the contribution of
ter (Millipore) before analysis. Each anthocyanin was identified on the different milled fractions to the total phenolics in brown rice.
the basis of retention time and the chromatography of authentic These data indicated that both the ratio of total phenolics in rice
standards. The percentage recoveries for Cy-3-G, Cy-3-R, and bran to that in polished rice and the ratio of phenolics in rice bran

Table 1
Total phenolic content and the percentage contributions of free and bound fractions to the total phenolics in milled fractions of black rice before and after extrusion.

Milled fraction Extrusion Phenolics (mg of GA equiv/100 g DW)

Free Bound Total
Rice bran Before 2945.0 ± 114.0 (85.8) 487.5 ± 11.4 (14.2) 3432.0 ± 106.0
After 3449.0 ± 17.2 (89.3)⁄ 414.8 ± 9.0 (10.7)⁄ 3863.0 ± 15.1⁄
Polished rice Before 80.1 ± 1.2 (75.8) 25.5 ± 0.7 (24.2) 105.7 ± 1.6
After 37.3 ± 0.9 (66.0)⁄ 19.2 ± 2.1 (34.0)⁄ 56.4 ± 2.4⁄
Brown rice Before 545.8 ± 3.0 (84.9) 97.2 ± 3.1 (15.1) 643.0 ± 6.1
After 111.9 ± 2.5 (60.4)⁄ 73.4 ± 1.9 (39.6)⁄ 185.3 ± 2.7⁄

Note: ⁄Denotes a significant difference between mean values before and after extrusion (p < 0.05), n = 3. Percentage of total phenolics shown in parentheses.
 H. Ti et al. / Food Chemistry 178 (2015) 186–194 189

to brown rice were both increased. In the unprocessed fractions, In the rice bran fraction, gallic, chlorogenic and syringic acids
the free, bound and total phenolic content in the rice bran were were detected either at low levels or not at all in the bound form.
5.4, 5.0 and 5.3 times higher than those in the brown rice, The free gallic acid content decreased significantly by 45.0% in the
respectively, and 36.8, 19.1 and 32.5 times higher than those in extruded fraction, while the free chlorogenic acid content
the polished rice, respectively. The free, bound and total phenolic increased significantly (p < 0.05) by 27.1% in the extruded fraction.
content in brown rice were 6.8, 3.8 and 6.1 times higher than those The free, bound and total syringic acid content increased by 61.4%,
in the polished rice, respectively. The rice bran fraction had the 8.2% and 58.7%, respectively, after extrusion. The vanillic and feru-
highest free, bound and total phenolic content followed by the lic acids were mainly distributed in the bound form with the free
brown rice, then the polished rice. In the extruded samples, form of vanillic acid decreasing by 35.6% and the bound form
the free, bound and total phenolic content in the rice bran were increasing by 1.9% after extrusion, but overall the total vanillic acid
30.8, 5.7 and 20.8 times higher than those in the brown rice, content after extrusion remained approximately the same. The free
respectively, and 92.5, 21.6 and 68.4 times higher than those in ferulic acid content decreased by 4.5% while in the extruded frac-
the polished rice, respectively. The free, bound and total phenolic tion, the bound and total amount of ferulic acid increased by
content in the brown rice were 3.0, 3.8 and 3.3 times higher than 14.1% and 13.7%, respectively. The caffeic and p-coumaric acids
those of the polished rice, respectively. were distributed in both the free and bound forms. After extrusion,
the free and bound caffeic acid content decreased by 39.5% and
5.5%, respectively, with the total content after extrusion being sig-
nificantly lower than that before extrusion. The free, bound and
3.2. Effect of extrusion on phenolics composition
total p-coumaric acid content increased by 14.7%, 16.2% and
15.7%, respectively, in the extruded fractions.
Seven phenolic compounds, gallic, protocatechuic, chlorogenic,
In the polished rice fractions, gallic, chlorogenic, caffeic and
caffeic, syringic, p-coumaric and ferulic acids, were detected in
syringic acids were found only in the free form. The amounts of
the free and bound forms in the unprocessed and extruded frac-
gallic, chlorogenic, caffeic and syringic acids decreased signifi-
tions of different milled black rice fractions (Fig. 1). The amounts
cantly (p < 0.05) by 23.0%, 65.5%, 30.0% and 60.5%, respectively,
of these seven compounds and the percentage contribution of free
after extrusion. Vanillic, p-coumaric and ferulic acids were present
and bound fractions to the total are presented in Table 2. Extrusion
mainly in the bound form. Compared with the unprocessed frac-
had no effect on the composition of these seven phenolic com-
tion, the free content of vanillic, p-coumaric and ferulic acids in
pounds in the free or bound forms in the rice bran, polished rice
the extruded fraction decreased by 41.6%, 57.9% and 50.8%, while
and brown rice fractions, but the content of these seven phenolics
the bound content increased by 4.6%, 9.1% and 5.5%, respectively.
were significantly different (p < 0.05).
Similar to what was observed for the bound forms, the total
amounts of vanillic, p-coumaric and ferulic acids changed very
COOH little after extrusion.
Gallic, chlorogenic, caffeic and syringic acids were only detected
in the free form in brown rice. The amounts of gallic, chlorogenic,
R1 R3 caffeic and syringic acids in the extruded fraction significantly
R2 decreased (p < 0.05) by 50.8%, 75.1%, 50.5% and 59.2%, respectively,
compared with those in the unprocessed fraction. The vanillic, p-
coumaric and ferulic acids were detected mainly in the bound
Gallic acid; R 1 =OH, R2 =OH, R3 =OH
Protocatechuic acid: R 1 =H, R 2=OH, R 3 =OH form. The free content of vanillic, p-coumaric and ferulic acids in
Vanillic acid: R1 =H, R 2 =OH, R3 =OCH 3 the extruded fraction were significantly lower than those in the
unprocessed fraction. The bound p-coumaric acid content
increased by 20.9% after extrusion but the amounts of bound vanil-
R3 COOR 4 lic and ferulic acids remained approximately the same. The total
amounts of vanillic and ferulic acids in the extruded fraction were
R2 similar to those in the unprocessed fraction but the total p-couma-
R1 ric acid content decreased by 10.2%.

Caffeic acid: R1 =OH, R2 =OH, R 3=OH, R 4=H

Coumaric acid: R 1=H, R 2=OH, R 3=H, R 4=H 3.3. Effect of extrusion on the total anthocyanin content
Ferulic acid: R 1=OCH 3, R2 =OH, R3 =H, R4 =H
Chlorogenic acid: R 1 =OH, R2 =OH, R3 =H, R4 =quinate The effects of extrusion on the free, bound and total anthocya-
nin content and the percentage contribution of each fraction to the
R1 total in different milled fractions of unprocessed and extruded
OH black rice flours are presented in Table 3. The data indicates that
+ anthocyanins existed mainly in the free form.
HO O In the rice bran fraction, extrusion caused a significant increase
(p < 0.05) of 5.5% in the free anthocyanin content, but a significant
R2 decrease (p < 0.05) of 30.8% in the bound anthocyanin content, but
R3 overall an increase (p < 0.05) of 5.4% in the total anthocyanin con-
tent. In the polished rice fraction, extrusion caused significant
decreases (p < 0.05) in free, bound and total anthocyanin content
Cyanidin 3-glucoside: R1 =OH, R2 =glucoside, R 3=OH of 89.3%, 40.6% and 88.4%, respectively. Similar to the polished rice
Cyanidin 3-rutinoside: R 1=OH, R 2 =rutinoside, R 3 =OH
Peonidin 3-glucoside: R1 =OCH 3 , R 2=glucoside, R 3=OH
fraction, in the brown rice fraction, extrusion caused almost com-
plete loss (p < 0.05) of 88.2%, 25.2% and 87.9%, of free, bound and
Fig. 1. Chemical structures of phenolic and anthocyanin compounds detected in total anthocyanin content, respectively. In the extruded black rice,
milled fractions of black rice before and after extrusion. the free fraction contained most of the total anthocyanin content
190 H. Ti et al. / Food Chemistry 178 (2015) 186–194

Table 2
Phenolic acid content and the percentage contributions of free and bound fractions to the total phenolics in different milled fractions of black rice before and after extrusion.

Milled fraction Extrusion Free Bound Total

Gallic acid Rice bran Before 42.4 ± 2.5 nd 42.4 ± 2.5
After 23.3 ± 1.0* nd 23.3 ± 1.0*
Polished rice Before 2.3 ± 0.1 nd 2.3 ± 0.1
After 1.8 ± 0.1* nd 1.8 ± 0.1*
Brown rice Before 17.3 ± 0.2 nd 17.3 ± 0.2
After 8.5 ± 0.2* nd 8.5 ± 0.2*
Chlorogenic acid Rice bran Before 7706.8 ± 377.2 nd 7706.8 ± 377.2
After 9799.1 ± 827.9* nd 9799.1 ± 827.9*
Polished rice Before 119.7 ± 2.4 nd 119.7 ± 2.4
After 41.3 ± 1.5* nd 41.3 ± 1.5*
Brown rice Before 1588.3 ± 83.0 nd 1588.3 ± 83.0
After 395.4 ± 12.1* nd 395.4 ± 12.1*
Vanillic acid Rice bran Before 59.2 ± 3.2 (6.4) 859.5 ± 36.6 (93.6) 918.7 ± 35.1
After 38.2 ± 0.5 (4.2)* 876.1 ± 23.9 (95.8) 914.3 ± 23.4
Polished rice Before 11.2 ± 1.0 (3.5) 308.9 ± 13.2 (96.5) 320.2 ± 12.9
After 6.62 ± 0.4 (2.0)* 323.3 ± 12.1 (98.0) 329.9 ± 11.6
Brown rice Before 17.3 ± 0.5 (5.8) 281.9 ± 13.7 (94.2) 299.2 ± 14.1
After 6.1 ± 0.5 (2.1)* 284.4 ± 10.8 (97.9) 290.6 ± 11.0
Caffeic acid Rice bran Before 30.3 ± 2.6 (61.2) 19.2 ± 0.5 (38.8) 49.6 ± 2.5
After 18.3 ± 1.6 (50.1)* 18.3 ± 1.7 (49.9) 36.7 ± 3.4*
Polished rice Before 1.0 ± 0.1 nd 1.0 ± 0.1
After 0.7 ± 0.0* nd 0.7 ± 0.0*
Brown rice Before 9.9 ± 0.2 nd 9.9 ± 0.2
After 4.9 ± 0.3* nd 4.9 ± 0.3*
Syringic acid Rice bran Before 181.2 ± 7.7 (94.9) 9.7 ± 0.5 (5.1) 190.9 ± 9.7
After 292.6 ± 9.2 (96.5)* 10.5 ± 0.3 (3.5) 303.0 ± 9.1*
Polished rice Before 7.6 ± 0.7 nd 7.5 ± 0.7
After 2.9 ± 0.0* nd 2.9 ± 0.1*
Brown rice Before 42.6 ± 5.5 nd 42.6 ± 5.5
After 17.4 ± 2.3* nd 17.4 ± 2.3*
Coumaric acid Rice bran Before 77.5 ± 2.9 (33.4) 154.4 ± 6.2 (66.6) 231.9 ± 6.1
After 89.0 ± 2.0 (33.2)* 179.3 ± 6.8 (66.8)* 268.3 ± 7.6*
Polished rice Before 1.8 ± 0.1 (22.2) 6.5 ± 0.8 (77.8) 8.4 ± 0.8
After 0.8 ± 0.1 (10.4)* 7.1 ± 0.4 (89.6) 7.9 ± 0.4
Brown rice Before 12.8 ± 0.3 (37.3) 21.5 ± 1.4 (62.7) 34.3 ± 0.6
After 4.7 ± 0.3 (15.6)* 26.0 ± 1.0 (84.4)* 30.8 ± 0.8*
Ferulic acid Rice bran Before 35.8 ± 2.2 (2.0) 1737.9 ± 104.3 (98.0) 1773.8 ± 102.1
After 34.2 ± 0.1 (1.7) 1981.9 ± 37.2 (98.3) 2016.2 ± 37.2*
Polished rice Before 5.9 ± 0.1 (7.2) 76.5 ± 5.7 (92.8) 82.5 ± 5.8
After 2.9 ± 0.2 (3.5)* 80.7 ± 1.0 (96.5) 83.6 ± 1.0
Brown rice Before 13.1 ± 0.5 (5.0) 249.9 ± 2.9 (95.0) 263.1 ± 2.9
After 3.5 ± 0.1 (1.5)* 237.7 ± 12.2 (98.5) 241.2 ± 12.1

Values expressed as lg/g DW (mean ± standard deviation, n = 3). Percentage of total phenolic acid content shown in parentheses.
*
Denotes a significant difference between mean values before and after extrusion (p < 0.05).

Table 3
Total anthocyanin content and the percentage contributions of free and bound fractions to the total anthocyanins in milled fractions of black rice before and after extrusion.

Milled fraction Extrusion Anthocyanin (mg of Cy-3-G equiv/100 g DW)

Free Bound Total
Rice bran Before 2109.0 ± 55.0 (99.6) 8.7 ± 0.5 (0.4) 2117.0 ± 55.0
After 2225.0 ± 35.0 (99.7)⁄ 6.0 ± 0.1 (0.3)⁄ 2231.0 ± 35.0⁄
Polished rice Before 19.7 ± 0.3 (98.4) 0.3 ± 0.0 (1.6) 20.0 ± 0.3
After 2.1 ± 0.2 (91.8)⁄ 0.2 ± 0.0 (8.2)⁄ 2.3 ± 0.2⁄
Brown rice Before 282.0 ± 1.9 (99.5) 1.5 ± 0.1 (0.5) 283.5 ± 2.0
After 33.2 ± 2.9 (96.8)⁄ 1.1 ± 0.04 (3.2)⁄ 34.3 ± 2.9⁄

Note: ⁄Denotes a significant difference between mean values before and after extrusion (p < 0.05), n = 3. Percentage of total anthocyanins shown in parentheses.

(91.8–99.7%), while the bound fractions only contributed a small 5.8 and 7.5 times higher, than those in the brown rice fractions and
proportion (8.2–0.3%) to the total. were 107.1, 29.0 and 105.9 times higher, respectively, than those
Similar to what was observed for the phenolic components, the in the polished rice fractions. The free, bound and total phenolic
contribution of different milled fractions to total anthocyanins in content in brown rice were 14.3, 5.0 and 14.2 times higher, respec-
brown rice changed significantly. Specifically, the data indicated tively, than those in the polished rice fractions. In the extruded
that the ratio of total anthocyanins in rice bran to that in polished fractions, the free, bound and total anthocyanin content in rice
rice and in brown rice were increased. In the unprocessed fractions, bran were 67.0, 5.5 and 65.0 times higher, respectively, than those
the free, bound and total anthocyanin content in rice bran were 7.5, in the brown rice fraction and 1059.5, 30.0 and 961.6 times higher,
 H. Ti et al. / Food Chemistry 178 (2015) 186–194 191

respectively, than those in the polished rice fraction. The free, In the brown rice fraction, compared with those in the unprocessed
bound and total anthocyanin content in brown rice were 15.8, fractions, the free, bound and total ORAC values decreased
5.5 and 14.9 times higher, respectively, than those in the polished (p < 0.05) by 16.7%, 9.2% and 14.7%, respectively.
rice fraction. The free, bound and total anthocyanin content were Extrusion treatment had slight effects on the contribution of
highest in rice bran, then in brown rice, followed by polished rice different milled fractions to the total antioxidant activity in brown
in the unprocessed and extruded black rice fractions. The free, rice. The data supported the fact that the proportions of different
bound and total phenolic content were significantly different in milled fractions changed only slightly after extrusion. In the unpro-
the extruded rice bran, brown rice and polished rice of black rice cessed fractions, the free, bound and total ORAC values in rice bran
compared with those in the unprocessed milled fractions were 5.1, 1.6 and 4.1 times higher, respectively, than those in the
(p < 0.05). brown rice fraction and 27.2, 7.5 and 21.8 times higher, respec-
tively, than those in the polished rice fraction. The free, bound
3.4. Effect of extrusion on anthocyanin composition and total ORAC values in brown rice were 5.4, 4.8 and 5.2 times
higher than those in the polished rice fractions, respectively. In
Cy-3-G, Cy-3-R and Pe-3-G (Fig. 1) were detected in the the extruded fractions, the free, bound and total ORAC values in
unprocessed and extruded fractions of different milled black rice rice bran were 7.3, 2.1 and 5.8 times the values in the brown rice
fractions (Table 4). In the black rice bran fraction, compared with fractions, respectively, and 51.7, 11.9, and 38.7 times the values
the unprocessed fraction, the Cy-3-G, Cy-3-R and Pe-3-G content in the polished rice fractions, respectively. The free, bound and
in the extruded fraction increased significantly (p < 0.05) by total ORAC values in brown rice were 7.1, 5.7 and 6.7 times higher
55.0%, 24.6% and 40.9%, respectively. In the polished rice fraction, than the values in the polished rice fractions, respectively. In the
the Cy-3-G, Cy-3-R and Pe-3-G content in the extruded fraction unprocessed fractions, the rice bran had the highest total ORAC
decreased significantly (p < 0.05) by 86.8%, 87.5% and 90.2%, values followed by the brown rice with the polished rice having
respectively, compared with those in the unprocessed fraction. the lowest total ORAC values. In the extruded fractions, the ORAC
Similarly, the Cy-3-G, Cy-3-R and Pe-3-G content in the brown rice value ordering was similar.
fraction after extrusion decreased significantly (p < 0.05) by 86.6%,
88.5% and 87.2%, respectively, compared with those in the unpro- 4. Discussion
cessed fraction.
4.1. Effect of extrusion on phenolics
3.5. Effect of extrusion on antioxidant activity
The total phenolics in unprocessed rice bran, polished rice and
The effects of extrusion on the free, bound and total antioxidant brown rice from black rice were 3432, 105.7 and 643.0 mg gallic
activity and the percentage contribution of each fraction to the acid equiv/100 g, respectively. These results suggest that the con-
total in different milled fractions of unprocessed and extruded centrations of phenolics were increased by the embryo in the rice
black rice flours are presented in Table 5. bran. The phenolic content in the rice bran fraction were 5.3 and
After extrusion, the free, bound and total ORAC values of rice 32.4 times higher than that in the corresponding brown and pol-
bran increased (p < 0.05) by 19.6%, 20.6% and 19.7%, respectively. ished rice fractions. This clearly supports previous reports that in
However, in the polished rice fraction, the free, bound and total rice, the phenolic content was mostly concentrated in the bound
ORAC values decreased (p < 0.05) by 37.0%, 23.3% and 33.1%, forms attached to cell wall materials (Adom & Liu, 2002). Shen,
respectively, compared with those in the unprocessed fractions. Jin, Xiao, and Bao (2009) reported that the total phenolic content

Table 4
Anthocyanin content in milled fractions of black rice before and after extrusion.

Milled fraction Extrusion Anthocyanins (mg/100 g DW)

Cy-3-G Cy-3-R Pe-3-G
Rice bran Before 527.6 ± 30.1 47.4 ± 5.5 219.9 ± 11.2
After 818.0 ± 53.7⁄ 62.9 ± 4.5⁄ 372.4 ± 22.9⁄
Polished rice Before 7.6 ± 0.0 0.8 ± 0.1 4.1 ± 0.1
After 1.0 ± 0.1⁄ 0.1 ± 0.0⁄ 0.4 ± 0.04⁄
Brown rice Before 87.9 ± 6.4 8.7 ± 0.2 42.1 ± 2.5
After 11.8 ± 0.6⁄ 1.0 ± 0.1⁄ 5.4 ± 0.4⁄

Note: ⁄Denotes a significant difference between mean values before and after extrusion (p < 0.05), n = 3.

Table 5
Total antioxidant activity and the percentage contributions of free and bound fractions to the total antioxidant activity in milled fractions of black rice before and after extrusion.

Milled fraction Extrusion ORAC (lmol Trolox equiv/ g DW)

Free Bound Total
Rice bran Before 587.5 ± 59.6 (90.1) 64.9 ± 18.0 (9.9) 652.4 ± 62.8
After 702.8 ± 60.3 (90.0)⁄ 78.3 ± 22.2 (10.0)⁄ 781.1 ± 59.5⁄
Polished rice Before 21.6 ± 4.9 (71.5) 8.6 ± 2.4 (28.5) 30.2 ± 5.4
After 13.6 ± 3.6 (67.3)⁄ 6.6 ± 1.7 (32.7) 20.2 ± 4.8⁄
Brown rice Before 116.2 ± 36.3 (73.7) 41.4 ± 9.1 (26.3) 157.6 ± 34.5
After 96.8 ± 14.5 (72.0)⁄ 37.6 ± 8.5 (28.0) 134.4 ± 17.7⁄

Note: ⁄Denotes a significant difference between mean values before and after extrusion (p < 0.5), n = 3. Percentage of total antioxidant activity shown in parentheses.
192 H. Ti et al. / Food Chemistry 178 (2015) 186–194

of six varieties of black rice grains ranged from 841 to 1245 mg gal- free phenolic acids during the extrusion process may in turn, pro-
lic acid equiv/100 g DW, with an average value of 1056 mg gallic mote polymerization of phenolics and tannins leading to reduced
equiv/100 g DW. It should be noted that these rice bran values extractability and antioxidant activity (Brennan, Brennan,
were obtained using only aqueous organic extracts (free forms of Derbyshire, & Tiwari, 2011; Joanne, David, & Len, 2000; Ragaee,
phenolics) of rice, while potentially significant amounts of bound Seetharaman, & Abdel-Aal, 2014). At the same time, bound pheno-
forms remained in the residue resulting in values that are signifi- lics are continuously released from the cell walls and could be
cantly lower than those found in the present study. transformed to free phenolics during the extrusion process. How-
The free phenolic content in rice bran, endosperm and whole ever, changes in the organizational structure of the extruded mate-
brown rice contributed 85.8%, 75.8% and 84.9% to the total, respec- rials may also lead to the increases in extractability. These factors
tively. This was consistent with the results of our previous study in each play a role during the extrusion process and affect each milled
rice bran from 12 black rice varieties (Zhang et al., 2010) where the fraction differently. Although the bound phenolic content in the
contribution of free phenolic content to the total ranged from polished and brown rice fractions decreased significantly
88.2% to 95.6% and those of Kong and Lee (2010) where in rice (p < 0.05), its contribution to the total phenolics in the extruded
bran, endosperm and whole grain from two Korean black varieties, samples was higher than in the unprocessed samples (Table 1).
the contributions were 91.2%, 71.1% and 90.1%, respectively. The The total content also decreased because of a lower level of bound
data from the present study also supported that the free phenolic phenolics in the polished and brown rice.
content existed mainly in the black rice bran/embryo fraction. This Whole brown rice consists of the starchy endosperm, germ and
fraction of black rice contributed most of the phenolics, which are bran (including aleurone), while rice bran (including the germ) is a
the primary source of health benefits. Therefore, this may partly good source of vitamins, phytochemicals, fiber and minerals that
explain the association between increased whole-grain consump- impart beneficial effects for human health. Some nutritional phyto-
tion and the reduced incidence of certain chronic diseases chemicals in rice bran exist mostly as glycosides, indicating they
(Jacobs, Meyer, Kushi, & Folsom, 1999; Jacobs, Slavin, & are linked to various sugar moieties, or they form other complexes
Marquart, 1995). by linking to carbohydrates, lipids, organic acids, amines and other
After extrusion, Table 1 clearly shows that the process caused phenols. Moreover, phytochemicals (phenolics and anthocyanins)
significant loss of the bound phenolic content. The percentage con- are commonly present in the bound form and as components of
tribution of free phenolics to the total was 89.3% in the final complex structures such as hydrolyzed tannins and lignins. Pheno-
extruded rice bran product, an increase of 3.5% compared with lic acids are sources of dietary phenols, which are linked to cellu-
the unprocessed material. Obviously, the extrusion process for lose, lignin, and proteins through ester bonds. Seven individual
raw black rice bran increased the percentage contribution of the phenolic acids (gallic, protocatechuic, chlorogenic, caffeic, syringic,
free phenolics to the total while decreasing the contributions from p-coumaric and ferulic acids) were detected in this study. Chloro-
the bound phenolics. The free phenolics may have increased genic and ferulic acids were the major phenolic acids observed in
because extrusion of the porous black rice bran leads to higher the black rice bran. Most of the ferulic acid was detected in the
extraction efficiency. At the same time, the cell wall friction dam- bound form, while chlorogenic acid was found only in the free
age during the extrusion process causes the release of bound phy- form. Adom and Liu (2002) reported that >93% of ferulic acids were
tochemicals. The breakdown of conjugated phenolics into free present in the bound form in corn, wheat, oats and rice, which
phenolics releases soluble proteins, fibers and polysaccharides. A agreed with the results of the present study. The extrusion treat-
similar increase has been observed in the total phenolics of cooked ment increased the total ferulic acid in rice bran and decreased
chickpeas, yellow peas, green peas and soybeans (Han & Baik, the free ferulic acid in the polished and brown rice. The total chlor-
2008). Nayak, Liu, Berrios, Tang, and Derito (2011) reported that ogenic acid content was higher in the extruded fraction than in the
extruded products prepared from purple potato and dry pea flours unprocessed fraction, mostly because of an increase in the free
had significantly higher phenolic content than the raw formula- form of the acid. Chlorogenic acid was not detected in the bound
tions. The contribution of free phenolics varied from 88% to 92% form likely because of decomposition aided by alkaline hydrolysis.
in the extruded products and from 69% to 84% in the raw samples. According to a previous report (Nayak et al., 2011), analysis of the
The increases in total phenolic content in black rice bran described change in the phenolic composition of rice suggested that it was
herein agreed with these previous reports. Additionally, our results caused by the breakdown of conjugated phenolics, the release of
also indicate that each milling fraction of whole black rice has a free phenolic acids and the formation of complex phenolic
different distribution of phenolics and they are modified by compounds.
extrusion.
In contrast, unlike rice bran, the bound phenolic content 4.2. Effect of extrusion on anthocyanins
decreased in the extruded polished and brown rice. Reductions
were also detected in the free and total phenolic content when Significant differences in anthocyanin content were observed
the polished and brown rice were extruded. The differences may between the different milled fractions. Free anthocyanins were
be due to the different phenolic compositions and content in the observed to be the dominant form in both the milled fractions
milled fractions of black rice. As reported herein, the phenolics in and the whole grain (brown rice). The contributions of the free
rice bran were 5.3 and 32.5 times higher, and the anthocyanins anthocyanins of rice bran, polished rice and brown rice, to the total
in rice bran were 7.5 and 105.9 times higher, respectively, than anthocyanins were 99.6%, 98.4% and 99.5%, respectively, indicating
those in brown and polished rice. The decreases in total phenolic that bound anthocyanins were only found in small amounts in all
content of corn and green beans were consistent with the trend fractions (Table 3). Free anthocyanins were found in large amounts
in polished and brown rice in our work (Jiratanan & Liu, 2004; in the black rice bran fraction. Kong and Lee (2010) found that the
Parra et al., 2007). Use of an extrusion thermal process will inevi- total anthocyanins in rice bran, polished rice and brown rice were
tably lead to a decrease of phenolics because of either phenolic 10.7, 0.2 and 1.9 mg Cy-3-G equiv/g DW, respectively. These values
compound decomposition caused by the high extrusion tempera- are obviously lower than the results presented herein, perhaps
ture or alterations in their molecular structure which may lead because of differences in the rice varieties and growing conditions
to a reduction in their chemical reactivity or a decrease in their between the two studies leading to significant differences in
extractability because of increased polymerization (Altan, anthocyanin content. After extrusion, the total anthocyanin con-
McCarthy, & Maskan, 2009). More specifically, decarboxylation of tent increased significantly in the black rice bran, while the total
 H. Ti et al. / Food Chemistry 178 (2015) 186–194 193

anthocyanin content decreased significantly in the polished and both before and after extrusion. The nutritional and health proper-
brown rice. This increase in the anthocyanins in the free black rice ties of black rice before and after its extrusion provide numerous
bran fraction may have come partly from the release of bound possibilities for its processing and use. Because of its decreased
anthocyanins during processing. The losses of anthocyanins in phytochemical content after extrusion, the polished rice fraction
the polished and brown rice may be attributed to the degradation is best suited to ordinary consumption. By comparison, the high
or decomposition of anthocyanins arising from thermal processing content of phenolics and anthocyanins in black rice bran and
(Hiemori, Koh, & Mitchell, 2009). Quantitative comparisons of brown rice both prior to and after extrusion makes them higher
anthocyanins before and after extrusion indicate that extrusion value rice products. Moreover, compared with other thermal pro-
had no significant effect on anthocyanin stability or retention. cessing methods, extrusion has less effect on the phytochemical
However, the amounts of anthocyanins were significantly different content of the raw materials because of the use of a shorter expo-
before and after extrusion. In black rice, anthocyanins are a group sure time to the high temperature. Therefore, the extrusion process
of natural colorants that are located in the aleurone layer. In a pre- may be useful for extensive processing of black rice in the food
vious study, Cy-3-G and Pe-3-G were identified as major antioxi- industry.
dant compounds in black rice (Hu, Zawistowski, Ling, & Kitts, The extrusion process can inevitably produce both positive and
2003). Our results also support the view that Cy-3-G and Pe-3-G negative influences on the bioactive compounds of the extrudates.
contribute the most to the anthocyanin content of black rice bran Some researchers reported that mild extrusion conditions (high
(Zhang et al., 2010), but less so in whole black rice (brown rice). moisture content, low residence time, low temperature) improved
Whole rice contains only minor amounts of Cy-3-R. The amounts the nutritional quality, while high extrusion temperatures (200 °C)
of Cy-3-G, Cy-3-R and Pe-3-G in the extruded fractions were signif- and low moisture content (<15%) can adversely impair nutritional
icantly higher than those in the unprocessed fractions (p < 0.05), quality (Singh, Gamlath, & Wakeling, 2007). Sharma et al. (2012)
which is contrary to the results that showed that decreases were found that among barley extrudates, the highest total phenolic
observed in black rice (Hiemori et al., 2009). Increases in the antho- content and antioxidant activity was exhibited by extrudates pre-
cyanin content of black rice bran extrudates could be attributed to pared using a low temperature and high moisture process. Regard-
the disruption of plant cell walls, thus resulting in better extract- less, controlling the temperature and moisture content are both
ability. The decreases of the predominant component, Cy-3-G, in necessary to prevent the loss of active substances and meet the
polished and brown rice support the results of Hiemori et al. needs of the food processing industry, such as pasting and curing.
(2009), which indicated that Cy-3-G is degraded into proto-
catechuic acid after heat treatment. 5. Conclusions

4.3. Effect of extrusion on antioxidant activity In conclusion, this study investigated the distributions of the
total phytochemicals (phenolics and anthocyanins) and antioxi-
Phenolic compounds contribute to antioxidant activity and dant activities in both free and bound forms of black rice. It also
reports examining free and bound antioxidant activities in rice explored how they contribute to the total content in the unpro-
bran, polished rice and whole black rice (brown rice) are limited. cessed and extruded milled fractions of black rice in terms of the
The percentage contribution of the free fraction in rice bran, pol- changes in the individual phenolic content. The total phenolics,
ished rice and whole black rice to the total ORAC values was anthocyanins and antioxidant activity in black rice bran increased
90.1%, 71.5% and 73.7%, respectively, which is similar to the contri- after extrusion processing, while the total phenolics, anthocyanins
butions of free phenolics to the total phenolic content. The bran and antioxidant activities in polished and brown rice were signifi-
fraction of black rice contributed the most to the antioxidant activ- cantly reduced by extrusion processing. This likely indicates that
ity. Our results indicate that black rice bran has a higher antioxi- the phenolics and anthocyanins were destroyed by extrusion treat-
dant activity than white rice bran. The free and bound ment. After extrusion, the milled fractions of black rice varied due
antioxidant activities in polished rice and whole black rice to differences in their phenolic composition and content. These
reported here using the ORAC assay could not be compared to val- results should serve as a starting point for developing different
ues in the literature because of the different analytical methods extruded fractions of whole black rice (brown rice) as products
used in previous reports. However, the total ORAC values of black with balanced nutritional value.
rice bran contained similar contributions from free (90.0%) and
bound (10.0%) forms as for the total phenolics in the extruded rice Acknowledgments
bran products. Increases in the total ORAC values followed the
same pattern as the total phenolics of the extruded products. This work was supported by the National Key Technology
Extrusion increased the free, bound and total ORAC values in black Research and Development Program for the 12th Five-year Plan
rice bran. The release of bound phytochemicals to free forms and (Nos. 2012BAD33B10 and 2012BAD33B08), the National ‘‘948’’
the formation of Maillard reaction products with antioxidant project (2014-Z52), the Special Fund for Public Welfare Industry
properties can lead to an increase in the total antioxidant activity. (Agriculture) (Research projects 201303071 and 201403063-2),
Previous studies reported that total ORAC values increased by 27– and the Innovation Capacity Construction Special Project of the
114%, 12–67% and 25–40%, respectively, in extruded green peas, Main Scientific Research Organization of Guangdong province
yellow peas and chickpeas (Xu & Chang, 2008), which is consistent (2011).
with the 19.7% increase in black rice bran observed in the present
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