Industrial Crops and Products 80 (2016) 141–147

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Industrial Crops and Products

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

Rosemary extract can be used as a synthetic antioxidant to improve

vegetable oil oxidative stability
Yong Yang a,b,1 , Xiaoxiao Song a,1 , Xiaonan Sui a,c,1 , Baokun Qi a,c , Zhongjiang Wang a,c ,
Yang Li a,c,∗ , Lianzhou Jiang a,c,∗
a
College of Food Science, Northeast Agricultural University, Harbin 150030, China
b
Key Laboratory of Processing Agricultural Products of Heilongjiang Province, College of Food and Bioengineering, Qiqihar University, Qiqihar 161006, China
c
National Research Center of Soybean Engineering and Technology, Harbin 150030, China

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

Article history: The protecting ability of rosemary extract as a plant-based antioxidant and an alternative to synthetic
Received 16 August 2015 antioxidants against the oxidation of vegetable oils was investigated in the present study. The effect
Received in revised form of rosemary extract on improving the oxidative stability of oils was evaluated using the Rancimat and
13 November 2015
Schaal oven tests. Oils with or without incorporation of rosemary extract or synthetic antioxidants were
Accepted 18 November 2015
analyzed using the Rancimat method at 120 ◦ C to determine their induction periods (IP). The changes
Available online 6 December 2015
in antioxidant capacity, total phenolic content, peroxide value, fatty acid composition, and tocopherol
content were measured under the Schaal oven test during storage at 62 ◦ C. The IP of oils incorporated with
Keywords:
Rosemary extract
rosemary extract was significantly (p < 0.05) higher than blank oils and oils with synthetic antioxidants.
Soybean oil The incorporation of rosemary extract into oils effectively prevented the oils by increasing the antioxidant
Cottonseed oil capacity and total phenolic content, decreasing the peroxide value, and delaying the degradation of
Rice bran oil tocopherols and polyunsaturated fatty acids of oils. Results of this study suggested the potential use of
Oxidative stability rosemary extract as an effective alternative to synthetic antioxidants.
Rancimat and Schaal oven tests © 2015 Elsevier B.V. All rights reserved.

1. Introduction
Lipid oxidation is one of the most critical factors affecting the
shelf life and the quality attributes of oil (da Silva and Jorge, 2014).
Lipid oxidation causes undesirable changes in taste, odor, texture,
flavor, and appearance of foods, and also destroys fat-soluble vita-
mins (Gallego et al., 2013; Zeb and Murkovic, 2013). Furthermore,
the oxidative degradation of lipids can damage biological mem-
branes, enzymes and proteins, which may pose a direct threat to
human health (Malheiro et al., 2013).
In recent decades, synthetic antioxidants, such as butylated
hydroxyanisole (BHA) and butylated hydroxytoluene (BHT), are
extensively used in retarding the oxidative degradation of lipids
due to their high oxidative stability and low cost. However, safety
concerns regarding the use of synthetic antioxidants have increased
globally. Synthetic antioxidants are inappropriate for chronically
ill patients, and the prolonged usage of synthetic antioxidants is
harmful to humans, potentially provoking the onset of degenera-
∗ Corresponding author. Fax: +86 451 55190716.
E-mail addresses: liyanghuangyu@163.com (Y. Li), jlzname@163.com (L. Jiang).
1
The first, second, and third author contributed equally to this work.
tive diseases (Cordeiro et al., 2013a,b; Xu et al., 2001). The benefits
of adding natural, plant-based antioxidants to lipids in order to pre-
vent lipid oxidation have been highlighted in recent years. Natural
antioxidants retard oxidative rancidity via the following pathways:
(1) capturing of free radicals; (2) decomposing/deoxidizing perox-
ides; and (3) scavenging oxygen (Zheng et al., 2012; Sun et al., 2010,
2011a).
Rosemary (Rosmarinus officinais L.) extract is popular as a natural
antioxidant due to its strong antioxidant capacity and fat-soluble
property, and has been adopted formally into the European regu-
lations (Commission Regulation (EU), 2011). The use of rosemary
extract as a natural antioxidant was first reported by Rac and Ostric-
Matijasevic in 1955 (Rac and Ostric-Matijasevic, 1955). Wu et al.
(1982) found that rosemary extract as a natural antioxidant had a
better antioxidant capacity than BHT and BHA. Chen et al. (1992)
reported that all rosemary extracts showed strong inhibitory effects
on lipid oxidation. The antioxidant capacity of rosemary extract
was attributed to the presence of phenolic diterpenes that scav-
enge singlet oxygen, hydroxyl radicals, and lipid peroxyl radicals,
thereby preventing lipid oxidation (Gallego et al., 2013; Zhang et al.,
2010; Sun et al., 2011a,b). Three types of phenolic diterpenes are
found in rosemary extract: carnosic acid, carnosol, and rosmanol;
with carnosic acid present as the major phenolic diterpene (Frankel

http://dx.doi.org/10.1016/j.indcrop.2015.11.044
0926-6690/© 2015 Elsevier B.V. All rights reserved.
142 Y. Yang et al. / Industrial Crops and Products 80 (2016) 141–147
et al., 1996; Thorsen and Hildebrandt, 2003). In addition, a previous
study indicated that carnosic acid was the most active antioxidant
component in rosemary extracts (Terpinc et al., 2009).
The Rancimat test has been widely used in evaluating the antiox-
idant capacities of synthetic and natural antioxidants. The Rancimat
method offers a real alternative to the active oxygen method for
the determination of oxidative stabilities owing to the apprecia-
ble saving in labor (Läubli and Bruttel, 1986). The Rancimat test
works by measuring the changes in conductivity caused by volatile
low-molecular weight organic acids, such as acetic acid and formic
acid, which are produced from oil oxidation under elevated tem-
peratures (Cordeiro et al., 2013a,b). The time that elapses until the
secondary oxidation products are produced is known as the induc-
tion period. This is used to characterize the resistance of oil to
oxidation, and the longer the induction time, the more stable the
oil is to oxidation.
The purpose of this study was to evaluate the effectiveness
of rosemary extract, compared to synthetic antioxidants (BHA
and BHT), on retarding the oxidation of three types of vegetable
oils–soybean oil, rice bran oil, and cottonseed oil, using the Ranci-
mat method and Schaal oven test.
2. Materials and methods
2.1. Materials
Soybean oil, rice bran oil, and cottonseed oil without added
antioxidants were provided by Hao Koufu Co., Ltd., Shanxi, China.
Commercial rosemary extract with a very high carnosic acid
content of 70% were purchased from Hainan Shupu Science
and Technology Ltd. Butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), and tocopherol standards (␣-tocopherol, ␥-
tocopherol, and ␦-tocopherol) were purchased from Sigma–Aldrich
(St Louis, MO, USA). All other chemicals and reagents used were of
analytical grade.
2.2. Rancimat analysis
The Rancimat analysis was conducted using a Rancimat 892
(Metrohm, Herisau, Switzerland) instrument. Rosemary extract
was directly added at a concentration of 400 mg/kg to the oils and
stirred for 10 min at room temperature. A mixture of synthetic
antioxidants (50% BHA and 50% BHT) was added at their legal limit
of 200 mg/kg (Duh and Yen, 1997) as a positive control. Oils with-
out added antioxidants were considered as blank controls. Oils (5 g
each) were accurately weighed into each reaction vessel. The tar-
get temperature was set at 120 ◦ C and airflow rate was 20 L/h. The
results were expressed as induction period (IP), which was auto-
matically determined from the inflection point of the curve using
the software supplied by the company.
2.3. Schaal oven storage stability test
In the processing and marketing of vegetable oils, the shelf life of
oils (period of time until an oil develops rancidity) is an important
quality factor. This is measured by its oxidative stability, and meth-
ods for evaluating oxidative stability of oils are therefore necessary.
One widely used method for evaluating oils is the Schaal oven test,
for which elevated storage temperatures was used to destroy the
original physicochemical attributes of oils (Antolovich et al., 2002).
The Schaal oven test was employed in the current study to evalu-
ate the effectiveness of rosemary extract in retarding the oxidative
deterioration of oils. Blank oils and oils incorporated with rosemary
extract and synthetic antioxidants (BHA + BHT) were accurately
weighed (50 g ± 0.01 g) into amber bottles without headspace and
stored in an oven at the constant temperature of 62 ± 1 ◦ C for 24
days, with samples taken every 6 days for DPPH, ABTS, total phe-
nolic content, and peroxide value analyses. Oil samples taken on
Day 0 and 24 were analyzed for their fatty acid composition and
tocopherol content.
2.4. DPPH and ABTS antioxidant assay
The DPPH antioxidant assay was performed according to Sui
and Zhou (2014). A spectrophotometer (UV Mini 1240, Shimadzu,
Kyoto, Japan) blanked using methanol was used in the analysis. A
volume of sample or Trolox standard (0.1 mL) was added into 3.9 mL
of DPPH stock solution (6 × 10−5 M), and left to stand for two hours
in the dark at room temperature. The absorbance of the mixture was
measured at the end of the two hours using the spectrophotometer
at 515 nm. Total antioxidant capacity measured using DPPH assay
was reported as mg Trolox equivalents per mL of sample.
The ABTS assay was performed following the procedure of Sui
et al. (2014). ABTS radical cation solution was produced by reacting
potassium persulfate (2.45 mM) with ABTS stock solution (7 mM)
in equal volumes, and the reaction was left to stand for 12–16 h
in the dark before use. ABTS radical cation solution (1 mL) was
diluted with 60 mL methanol to achieve an absorbance reading of
0.70 (±0.02) at 734 nm. An aliquot of sample or Trolox (2.5 ␮L) was
allowed to react with the diluted ABTS radical cation solution (1 mL)
for seven minutes before measuring its absorbance at 734 nm using
the spectrophotometer. The total antioxidant capacity obtained
using the ABTS assay was reported as mg Trolox equivalents per
mL of sample.
2.5. Peroxide value measurement
The peroxide value (PV) indicates the state of primary oil oxi-
dation. The factors that result in a high PV value are the history of
oil and exposure to factors that cause oxidation, such as high tem-
peratures during production and storage (Hui and Chandan, 2007).
The peroxide value was determined according to our previous work
(Li et al., 2014). Oil samples (300 mg) were accurately weighed and
dissolved in 9.9 mL of chloroform/methanol mixture (7:3, v/v) fol-
lowed by the addition of 50 mL of xylenol orange (10 mM) and 50 ␮L
of iron (II) chloride solution. The mixture was left to react for 5 min
at room temperature before centrifuging at 1000 g for 5 min. The
absorbance was measured at 560 nm using the spectrophotometer.
Results were expressed in miliequivalents of active oxygen per kg
of oil.
2.6. Total phenolic content measurement
The total phenolic content of oils was measured using the
Folin–Ciocalteau method according to the work of Zheng et al.
(2012) with some modifications. A 100 mg aliquot of oil sample was
mixed with the Folin–Ciocalteau reagent (0.5 mL) and methanol
(2 mL). The mixture was shaken and 1.5 mL of 15% Na2 CO3 was
added into the mixture followed by shaking the mixture for 30s.
Distilled water was added into the mixture to make a final vol-
ume of 7 mL. The mixture was then incubated at 50 ◦ C for 20 min
before centrifuging at 2000 g for 10 min. After centrifugation, the
supernatant was transferred into a cuvette and its absorbance mea-
sured at 750 nm. The total phenolic content of the oil samples was
expressed as gallic acid equivalents.
2.7. Fatty acid composition analysis
The fatty acid composition was analyzed using an Agilent 7890
gas chromatograph coupled with an Agilent 5975 mass spec-
trometer (GC–MS; Agilent Technology, CA, USA), and equipped
with an HP-88 capillary column (100 mm × 0.25 mm id, 0.2 ␮m
 Y. Yang et al. / Industrial Crops and Products 80 (2016) 141–147 143

Table 1 synthetic antioxidants, indicating that rosemary extract was more

Induction period of the three types of oils obtained from the Rancimat test.
Oil samples Induction period (h)
Blank Rosemary extract
Soybean oil 2.2 ± 0.22c 3.4 ± 0.18a
Cottonseed oil 1.88 ± 0.2c 3.35 ± 0.15a
Rice bran oil 3.83 ± 0.07c 6.22 ± 0.21a
Mean values in the same row followed by the same superscript letters are not
significantly different (p > 0.05).
film thickness). Analysis of fatty acid composition was performed
according to our previous study (Li et al., 2013). To prepare fatty
acid methylesters (FAME), oils were saponified with 0.5 M KOH and
then methylated using 40% BF3 in methanol. The injection tem-
perature was set to 250 ◦ C. Helium was used as a carrier gas at a
pressure of 100 kPa. A split ratio of 1:30 was used. The oven tem-
perature was programmed as such: 80 ◦ C for 5 min, followed by an
increased to 150 ◦ C at 10 ◦ C/min, then held at 150 ◦ C for 2 min, fol-
lowed by another increase to 230 ◦ C at 5 ◦ C/min, and finally held at
230 ◦ C for 10 min. The ionization voltage was 70 eV, and a scanning
range of 50–550 m/z was used. Each fatty acid was quantified using
external standards.
2.8. Measurement of tocopherols
The tocopherol content was measured according to Bele et al.
(2013). Oil samples were diluted in a solution prepared using
methanol, hexane, and tetrahydrofuran at a volumetric ratio of
80:10:10. The mixture was vortexed and centrifuged, and an
aliquot of the supernatant was injected into a Waters 2695 Alliance
high performance liquid chromatography system, equipped with a
C18 column (Sunfire, 250 × 4.6 mm i.d., 5 ␮m; Waters, Wexford,
Ireland). A mixture containing acetonitrile and methanol (50:50)
was used as a mobile phase at a flow rate of 1 mL/min. Quantita-
tion of tocopherols was performed using a fluorescence detector at
290 nm excitation wavelength and 325 nm emission wavelength.
effective in stabilizing oil against oxidative deterioration compared
to synthetic antioxidants.
BHA + BHT
3.2. The antioxidant capacity of oils during storage
2.9 ± 0.12b
2.84 ± 0.13b
4.79 ± 0.18b The measurement of antioxidant capacity of oils is a widely used
method for the determination of the stability of vegetable oils (Zeb
and Murkovic, 2013). The change in antioxidant capacity of oils
measured by both DPPH and ABTS assays during storage is shown
in Fig. 1A and B and analyzed using ANOVA (data for day 6, 12, 18 is
not shown as no large difference between them). Initially, oils with
rosemary extract incorporated showed higher antioxidant capac-
ity than blank oils and oils with synthetic antioxidants (BHA + BHT)
incorporated. During the 24-day storage, the antioxidant capacity
of blank oils and oils with rosemary extract or synthetic antiox-
idants incorporated decreased significantly. On Day 24, soybean
oil and cottonseed oil with rosemary extract incorporated showed
significantly higher antioxidant capacity than that of oil with syn-
thetic antioxidants. However, the antioxidant capacity of rice bran
oil with either rosemary extract or synthetic antioxidants incorpo-
rated was not statistically different at the end of storage.
Carnosic acid is the major active compound in rosemary extract
and also the most active antioxidant component (Terpinc et al.,
2009). The chemical structure of carnosic acid shows that it has
two O-phenolic hydroxyl groups located at the benzene ring, and
in comparison BHA and BHT only have one (Fig. 1). Although the
antioxidant capacity of a compound is usually determined by the
number of O-phenolic hydroxyl groups found in its molecule, the
antioxidant capacity of a compound could also be affected by the
position of O-phenolic hydroxyl groups and the presence of other
functional groups in its molecular structure. Carnosic acid contains
two adjacent O-phenolic hydroxyl groups strained with an adjacent
carboxylic group (-CO2 H), which gives rise to its strong antioxidant
ability (Erkan et al., 2008).
3.3. Total phenolic content

2.9. Statistical analysis
All sampling and chemical analyses were performed in tripli-
cate. Results of this study were expressed as a mean of the triplicate
values with standard deviations. Statistical analysis was conducted
on the obtained results using one-way analysis of variance (ANOVA)
analysis and Duncan’s multiple range test with the help of SPSS
software (SPSS Inc., USA).
3. Results and discussion
3.1. Rancimat analysis
The Rancimat analysis aimed to measure the induction period
(IP) by detecting the formed volatile acids during oil oxidation
(Mathäus, 1996). The Results showing the effect of rosemary extract
and synthetic antioxidants (BHA + BHT) on the oxidative stability
of vegetable oils are shown in Table 1. Unsurprisingly, oils with-
out the addition of any antioxidants were the most easily oxidized
samples, as indicated by with the lowest IP values obtained. The IP
values of oils with added rosemary extract or synthetic antioxidants
were found to be significantly larger (p < 0.05) than that of the blank
oils. Initially, the rice bran oil showed a noticeable larger IP value
(3.83 h) than soybean (2.2 h) and cottonseed oil (1.88 h). This was
due to the presence of gamma oryzanol in rice bran oil, which has
been reported to have a very strong antioxidant capacity (Xu et al.,
2001). The IP values of the three types of oils with rosemary extract
incorporated were significantly higher than that of oil with added
The total phenolic content of oils was examined through 24 days
of storage (Fig. 1C) and analyzed using ANOVA labeled with lower
and uppercase letters (data for day 6, 12, 18 is not shown as no
large difference between them). The blank rice bran oil contained a
much higher total phenolic content (502.64 mg gallic acid equiv-
alents/kg sample) than soybean and cottonseed oil initially. On
Day 0, all three types of oils with rosemary extract incorporated
exhibited significantly (p < 0.05) higher total phenolic content than
their corresponding blank oils and oils with synthetic antioxidants
(BHA + BHT) incorporated. During storage, the total phenolic con-
tent of all oils with or without added antioxidants decreased with
increasing storage time, and had decreased significantly by Day 24.
Nevertheless, oils with rosemary extract incorporated still showed
the highest total phenolic content after storage. The decrease in
total phenolic content may be caused by the decomposition and
oxidation of phenolic compounds in oils which undergo qualita-
tive and quantitative modifications during storage (Gargouri et al.,
2014). In addition, the phenolic compounds in oils may act as
an antioxidant by donating H-atom(s) to free radicals which con-
tributes to its decrease. (Samaniego Sánchez et al., 2007).
3.4. Peroxide value analysis
Peroxides are the main initial products of oil oxidation, and
can be determined using the peroxide value (PV) (Malheiro et al.,
2013). A higher peroxide value implies a lower oxidative stability
(Naghshineh et al., 2010). The changes in PV values were shown in
Table 2. During the 24-day storage, PV values of all the oils with
144 Y. Yang et al. / Industrial Crops and Products 80 (2016) 141–147

Fig. 1. Changes in the antioxidant capacity measured by DPPH (A) and ABTS (B) assays, and total phenolic content (C) during storage. Letters in lower case denoted significant
differences (p<0.05) among blank oils and oils with rosemary extract or synthetic antioxidants incorporated on Day 0 and 24 (highlighted in green range), and letters in upper
case denoted significant difference for each type of oil on Day 0 or 24 (highlighted in blue range).

Table 2
Change in the peroxide value (meq/kg) of oils during storage.

Oil sample Storage time (days) Peroxide value (meq/kg)

Blank Rosemary extract BHA + BHT

Soybean oil 0 1.46 ± 0.09 a

1.45 ± 0.11
a
1.43 ± 0.04a
6 8.57 ± 0.09a 3.42 ± 0.09c 4.27 ± 0.14b
12 15.93 ± 0.13a 7.38 ± 0.11c 6.32 ± 0.16b
18 19.76 ± 0.21a 12.16 ± 0.17c 15.42 ± 0.22b
24 23.72 ± 0.51a 17.32 ± 0.15c 19.50 ± 0.20b
Cottonseed oil 0 4.80 ± 0.12a 4.47 ± 0.15b 4.60 ± 0.17ab
6 9.68 ± 0.13a 6.30 ± 0.17c 7.62 ± 0.12b
12 15.80 ± 0.13a 7.52 ± 0.14c 10.53 ± 0.15b
18 23.81 ± 0.12a 12.59 ± 0.13c 14.81 ± 0.16b
24 19.47 ± 0.18a 16.53 ± 0.24b 19.75 ± 0.66a
Rice bran oil 0 3.53 ± 0.23b 3.59 ± 0.15b 4.28 ± 0.17a
6 9.53 ± 0.14a 8.17 ± 0.14b 8.40 ± 0.15b
12 16.71 ± 0.16b 14.93 ± 0.10c 17.79 ± 0.15a
18 25.61 ± 0.13a 17.68 ± 0.16c 19.87 ± 0.13b
24 29.45 ± 0.61a 19.00 ± 0.19c 21.56 ± 0.23b

Mean values in the same row for each oil followed by the same superscript letters are not significantly different (p > 0.05).

and without added antioxidants increased sharply, except for the
blank cottonseed oil. The PV values of blank cottonseed oil showed
a decrease on Day 24 after the constant increase from Day 0 to
Day 18. The drop in the PV values was due to the unstable pri-
mary oxidation products (hydroperoxides) that are susceptible to
decomposition, forming carbonyl compounds (Shahidi and Zhong,
2010). This indicated the poor oxidative stability of blank cotton-
seed oil, which correlates with its short induction period indicated
by the Rancimat test.
On Day 24, the blank soybean oil and rice bran oil had sig-
nificantly (p < 0.05) higher PV values than their corresponding
oils with rosemary extract and synthetic antioxidants (BHA + BHT)
incorporated. In comparison, the PV values of blank cottonseed
oil (19.47 meq/kg) and cottonseed oil with synthetic antioxidants
(19.75 meq/kg) at the end of 24 days of storage were not signifi-
cantly different. The rosemary extract exhibited a greater ability
to retard the PV values of oils, i.e., oils with rosemary extract
incorporated showed significantly lower PV values than that of
oils with synthetic antioxidants incorporated. More specifically,
the PV values of soybean oil with rosemary extract and syn-
thetic antioxidants incorporated were 17.32 and 19.50 meq/kg,
respectively, and of cottonseed oil were 16.53 and 19.75 meq/kg,
respectively, and of rice bran oil were 19.00 and 21.56 meq/kg,
respectively.
3.5. Tocopherol content analysis
The tocopherol content in oils on Day 0 and 24 is shown in
Table 3. Tocopherols are monophenolic compounds that are often
found in edible oils (Choe and Min, 2009). They are highly soluble in
oil and are the most important antioxidants in oils. Among the three
types of oils, cottonseed oil was found to contain the most abun-
dant amount of total tocopherols (136.2 mg/100 mL), followed by
the soybean oil containing 95.03 mg/100 mL of total tocopherols.
However, the tocopherol content in rice bran oil was very low,
which is in agreement with published data (Posuwan et al., 2013).
After the 24-day storage, the tocopherol content in oils decreased
significantly. For blank soybean oil and cottonseed oil, their total
tocopherol content decreased from 95.03 and 136.2 mg/100 mL to
13.96 and 62.27 mg/100 mL respectively, while for blank rice bran
oil no tocopherols could be detected after storage. The incorpo-
ration of rosemary extract in oils slowed the rate of degradation
of tocopherols. In comparison, synthetic antioxidants (BHA + BHT)
exhibited a poorer ability to prevent degradation of tocopherols.
At the end of storage, individual tocopherols (␣-tocopherol, ␥-
tocopherol, and ␦-tocopherol) and total tocopherol content of oils
with rosemary extract incorporated were significantly higher than
oils with synthetic antioxidants incorporated. Chotimarkorn and
Silalai (2008) reported a similar decrease in tocopherol content
Table 3
Tocopherol content (mg/100 mL) of the three types of oils on Day 0 and 24* .

Tocopherols Soybean oil Cottonseed oil Rice bran oil

Blanka Blank ORa OSa Blank Blank OR OS Blank Blank OR OS

Day 0 Day 24 Day 24 Day 24 Day 0 Day 24 Day 24 Day 24 Day 0 Day 24 Day 24 Day 24

␣-Tocopherol 5.56 ± 1.05a nd 2.52 ± 0.72b nd 2.69 ± 0.41a nd nd nd 16.65 ± 0.95a nd 4.57 ± 1.56b 3.16 ± 0.23b
␥-Tocopherol 66.2 ± 2.76a 13.96 ± 0.25d 39.61 ± 1.54b 25.37 ± 3.29c 82.86 ± 1.17a 40.16 ± 2.18d 60.23 ± 3.89b 49.54 ± 2.11c 4.25 ± 1.02a nd 0.87 ± 0.77b nd
␦-Tocopherol 23.33 ± 3.04a nd 10.36 ± 1.50b 5.28 ± 1.01c 50.66 ± 0.76a 22.11 ± 2.27d 36.38 ± 2.84b 31.31 ± 1.52c nd nd nd nd
Total content 95.03 ± 1.25a 13.96 ± 0.25d 52.5 ± 2.81b 30.64 ± 2.96c 136.2 ± 2.03a 62.27 ± 2.37d 96.61 ± 6.55b 80.84 ± 1.22c 20.9 ± 0.77a nd 5.44 ± 1.03b 3.16 ± 0.21c

Y. Yang et al. / Industrial Crops and Products 80 (2016) 141–147

a
Blank, oil without antioxidant; OR, oil incorporated with rosemary extract; OS, oil incorporated with synthetic antioxidants (BHA + BHT).

Table 4
Fatty acid composition of the three types of oils on Day 0 and 24* .

Fatty acids Soybean oil Cottonseed oil Rice bran oil

Blanka Blank ORa OSa Blank Blank OR OS Blank Blank OR OS

Day 0 Day 24 Day 24 Day 24 Day 0 Day 24 Day 24 Day 24 Day 0 Day 24 Day 24 Day 24

C14:0b 0.08 ± 0.01b 0.29 ± 0.01a 0.09 ± 0.01b 0.17 ± 0.17ab 0.64 ± 0.06c 0.85 ± 0.10a 0.71 ± 0.05bc 0.78 ± 0.05ab 0.26 ± 0.01b 0.35 ± 0.07a 0.29 ± 0.02ab 0.3 ± 0.02ab
C16:0 8.73 ± 0.14d 17.65 ± 0.20a 10.13 ± 0.09c 11.34 ± 1.10b 22.1 ± 0.34c 39.04 ± 0.73a 28.06 ± 0.45b 30.84 ± 1.72b 17.18 ± 0.25c 19.24 ± 0.27a 17.64 ± 0.16b 17.91 ± 0.18b
C18:0 1.75 ± 0.07d 4.33 ± 0.20a 2.23 ± 0.11c 2.62 ± 0.19b 1.65 ± 0.09d 7.85 ± 0.06a 3.84 ± 0.08c 4.38 ± 0.14b 1.52 ± 0.15c 3.47 ± 0.25a 1.75 ± 0.05bc 1.89 ± 0.08b
C18:1 25.49 ± 0.38b 34.51 ± 10a 36.18 ± 1.27a 36.62 ± 1.36a 16.4 ± 0.42c 19.01 ± 1.46b 20.8 ± 0.74a 18.96 ± 0.78b 43.35 ± 1.11b 46.03 ± 0.6a 44.7 ± 0.96ab 43.99 ± 0.71b
C18:2 51.75 ± 0.3a 30.76 ± 2.01c 44.62 ± 0.35b 41.47 ± 1.34b 51.5 ± 0.59a 33.31 ± 1.27c 40.32 ± 1.1ab 37.72 ± 0.52b 35.41 ± 2.07a 29.67 ± 1.15b 35.38 ± 1.52a 32.24 ± 1.26b
C18:3 6.29 ± 0.21a 1.04 ± 0.12d 5.7 ± 0.30b 3.1 ± 0.13c 3.98 ± 0.50a 0.96 ± 0.50c 3.14 ± 1.30ab 2.23 ± 0.83b 1.23 ± 0.12a 0.58 ± 0.15c 1.05 ± 0.16ab 0.88 ± 0.14b
C20:0 0.79 ± 0.10a 0.97 ± 0.10a 0.82 ± 0.09a 0.86 ± 0.08a 0.35 ± 0.07d 1.43 ± 0.18a 0.68 ± 0.10c 0.98 ± 0.17b 0.71 ± 0.09b 1.56 ± 0.22a 0.77 ± 0.14b 0.88 ± 0.11b
SFA 11.35 ± 0.33c 23.24 ± 0.61a 13.27 ± 0.40bc 14.99 ± 1.55b 24.74 ± 0.57c 48.17 ± 0.98a 33.29 ± 0.70bc 36.98 ± 2.09ab 19.67 ± 0.51b 24.62 ± 0.82a 20.45 ± 0.38b 20.98 ± 0.40b
MUFA 25.49 ± 0.38b 34.51 ± 10a 36.18 ± 1.27a 36.62 ± 1.36a 16.4 ± 0.42c 19.01 ± 1.46b 20.8 ± 0.74a 18.96 ± 0.78b 43.35 ± 1.11b 46.03 ± 0.60a 44.7 ± 0.96ab 43.99 ± 0.71b
PUFA 58.04 ± 0.52a 31.8 ± 2.13d 50.32 ± 0.76b 44.57 ± 1.48c 55.48 ± 1.2a 34.27 ± 1.78c 43.46 ± 2.38a 39.95 ± 1.35b 36.64 ± 2.30a 30.25 ± 1.31c 36.43 ± 1.69a 33.12 ± 1.41b
a
Blank, oil without antioxidant; OR, oil incorporated with rosemary extract; OS, oil incorporated with synthetic antioxidants (BHA + BHT).
b
C14:0, myristic acid; C16:0, palmitic acid; C18:0, stearic acid; C18:1, oleic acid; C18:2, linoleic acid; C18:3, linolenic acid; C20:0, arachidic acid.

145
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when evaluating dough fried with soybean and rice bran oil during
a 10-day storage period at 60 ◦ C.
3.6. Fatty acid composition analysis
The changes in fatty acid composition reflected the oxidative
stability and nutritional quality of oils. The fatty acid composition
of oils with and without antioxidants on Day 0 and 24 is shown
in Table 4. The three oils contained variable amounts of individual
fatty acids and as well as saturated fatty acids (SFAs), monounsatu-
rated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs).
Linoleic acid (C18:2) was the dominant fatty acid of soybean oil
(51.75%) and cottonseed oil (51.5%), while rice bran oil contained
slightly more oleic acid (C18:1, 43.35%) than linoleic acid (35.41%).
The initial SFA content of soybean oil (11.35%) was much lower
than that of cottonseed oil (24.74%) and rice bran oil (27.41%). The
levels of C14:0, C16:0, C18:0, and C20:0 increased in all oils on Day
24, and some increases were significant while some were not. The
level of C18:1, C18:2 and C18:3 had increased for the three types
of oils with/without antioxidants. The incorporation of rosemary
extract in oils helped to preserve the integrity of the unsaturated
fatty acids, such as C18:1, C18:2 and C18:3. Although the synthetic
antioxidants (BHA + BHT) also had the same effect, they were less
effective.
After storage, the SFA contents of blank soybean oil (23.24%),
cottonseed oil (48.17%), and rice bran oil (24.62%) were twice as
high compared to their initial values. However, in oils with rose-
mary extract or BHA + BHT incorporated, the SFA contents of the
three types of oils were only increased slightly. Although not statis-
tically significant, rosemary extract was shown to be slightly more
effective in lowering the SFA contents than BHA + BHT. Interest-
ingly, on Day 24, the MUFA contents of all the oils had increased.
Sun-Waterhouse et al. (2011) also observed increased MUFA con-
tents in encapsulated oil with and without the incorporation of
caffeic acid stored at both 20 and 37 ◦ C, in their study evaluating
the stability of encapsulated olive oil with caffeic acid. The unsatu-
rated fatty acids of oil are known to be easily oxidized (Gargouri
et al., 2014; Marfil et al., 2008). An increase in the number of
double bonds or electron donating groups in unsaturated fatty
acids leads to a decrease in the oxidative stability (Choe and Min,
2006). At the end of storage, the amount of PUFA was significantly
reduced in all blank oils and oils with BHA + BHT incorporated.
However, the incorporation of rosemary extract into cottonseed
and rice bran oil did not significantly change their PUFA levels
(p > 0.05). On the other hand, the PUFA content in soybean oil with
rosemary extract incorporated decreased slightly from 58.04 to
50.32%, but was still significantly higher than that of soybean oil
with synthetic antioxidants (44.57%). The incorporation of rose-
mary extract into vegetable oils may be a feasible and effective
approach to protect the oil from oxidation during storage. The
incorporation of rosemary extract further offers oil and food indus-
tries a new possibility to preserve unsaturated fatty acids. Besides,
the incorporation of rosemary extract not only provided additional
protection to vegetable oils, but also improved their nutritional
values.
4. Conclusion
The effectiveness of rosemary extract in protecting oil from
oxidative rancidity was evaluated in this study. Results showed
that, in comparison with synthetic antioxidants, rosemary extract
was more effective in delaying the oxidation of the three types of
oils. The oils with rosemary extract incorporated exhibited much
better chemical stability, including higher induction period, antiox-
idant capacity, total phenolic content, lower peroxide value, and
slightly decreased tocopherols content and PUFAs. Therefore, the
use of rosemary extract could be an effective and way to protect
oils from oxidation.
Acknowledgements
The authors would like to acknowledge the support for this
study by the National Natural Science Foundation of China
(research grant number: 31430067 and 31571876), National
Key Technology Support Program (research grant number:
2014BAD22B01-02), Natural Science Foundation of Heilongjiang
Province of China (research grant number: ZD201302 and
C201331), Agricultural Key Technologies R & D Program of Qiqihar
Science and Technology Bureau (research grant number: NYGG-
201206-3) , and the Key Laboratory of Soybean Biology of Chinese
Education Ministry.
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