104 2012, Vol. 33, No.

04 食品科学 ※工艺技术

Ultrasound-Assisted Extraction and Heat Stability of Capsaicinoids from

Doubanjiang (Bean Paste)
JIA Hong-feng1，PENG De-chuan1，LIANG Ai-hua 1，HE Jiang-hong1，CHEN Zu-ming1，HE Zhi-fei2
(1. Department of Food Science, Sichuan Higher Institute of Cuisine, Chengdu 610100, China；
2. College of Food Science, Southwest University, Chongqing 400716, China)

Abstract ：A fast method has been developed for the extraction of capsaicinoids from Doubanjiang employing ultrasound-
assisted extraction (UAE). The effects of operating parameters such as solvent type, temperature, extraction time and number
of repeated extractions on the extraction rates of capaicin, dihydrocapsaicin and capsaicinoids were investigated. The optimum
extraction conditions were ultrasound power of 200 W, ultrasound frequency of 40 kHz, Doubanjiang/methanol ratio of 2.5:10
(g/mL), temperature of 70 ℃, and twice-repeated extractions for 5 min each time. The results of thermal stability experiments
indicated that capsaicinoid contents in Doubanjiang began to gradually decrease as the temperature increased from 130 ℃ and
exhibited a distinct decrease when the temperature was over 200 ℃.
Key words：Doubanjiang；ultrasound-assisted extraction；capsaicinoids；thermal stability；high performance-liquid
chromatography (HPLC)

豆瓣中辣椒素类物质的超声波提取及其热稳定性

贾洪锋 1 ，彭德川 1 ，梁爱华 1 ，何江红 1 ，陈祖明 1 ，贺稚非 2

(1.四川烹饪高等专科学校食品科学系，四川 成都 610100；2.西南大学食品科学学院，重庆 400716)

摘 要：对豆瓣中辣椒素类物质的超声波提取条件(提取溶剂、提取温度、提取时间和提取次数)和豆瓣中辣椒素类
物质的热稳定性进行研究。结果表明：最佳提取条件为超声波功率 200W、超声波频率 40kHz、2.5g 豆瓣采用 10mL
甲醇提取两次、提取温度 70℃、提取时间 5min；热稳定性实验表明，随着加热温度的升高，豆瓣中辣椒素类物
质的含量会逐渐下降，当温度达到 130℃时，豆瓣中辣椒素类物质的含量开始下降，当温度达到 200℃时，辣椒
素类物质的含量明显下降。
关键词：豆瓣；超声波提取；辣椒素；热稳定性；高效液相色谱
中图分类号：TS201.1；TS207.3 文献标识码：A 文章编号：1002-6630(2012)04-0104-05

Douban is a kind of fermented seasoning produced
using broad bean, hot pepper and some condiments for cui-
sine use in China. It attributes food of pungency and special
flavor when cooking. The major pungent compounds in
Douban are capsaicin and dihydrocapsaicin, they are respon-
[1-2]
sible for about 90% of the pungency . The techniques for
[3-6]
capsainoids extraction are maceration , magnetic stirring
Soxhlet[9], microwave-assisted extraction[10-11], supercritical
[12] [6,13]
CO2 extraction , and ultrasound-assisted extraction
The application of ultrasound-assisted extraction (UAE)
offers many advantages including the reduction of solvents,
temperature and the time for extraction, which is very useful
for the extraction of thermolabile and in unstable compounds[13].
While many papers have been published dealing with the
UAE of different plant materials[14-17]. Some studies reported
that heating temperature and the time had a remarkable influ-
[7-8]
, ence on capsaicinoids content in peppers[10,18]. However, the
influence of heating temperature and the time on
. capsaicinoids content in Douban has not been reported. In
this experiment, an ultrasound-assisted extraction method

收稿日期：2011-03-21
基金项目：四川省教育厅科研项目(10ZB084)；四川省教育厅川菜发展研究中心科研项目(CC09Z01)
作者简介：贾洪锋(1981 —)，男，讲师，硕士，研究方向为食品分析与检测。E-mail：jiahongfeng_cq@163.com
※工艺技术 食品科学
was optimized to extract the capsaicin and dihydrocapsaicin
from Douban, and the influence of heating temperature and
the time on capsaicinoids content in Douban had been
studied.
1 Materials and Methods
1.1 Materials, reagents and instruments
For the Douban sample, Linjiangsi ,, Douban was
,,
purchased from local retail markets in Chengdu, China.
Capsaicin (95%) and dihydrocapsaicin (90%) were
obtained from Sigma Chemical Co (St. Louis, MO, USA);
Methanol (HPLC-grade) was purchased from Fisher Chemi-
cal Co (Fairlawn, MA, USA); Ethanol (HPLC-grade) and ac-
etone (AR-grade) were purchased form Kelong Chemical Co
(Chengdu, China).
Ultrasonic bath (Shumei, KQ5200E, Jiangsu, China)；
Purkinje liquid chromatograph model L9 (Purkinje general,
Beijing, China) was consisted of a sample injector (Hamilton
710SNR 100μL SYR), a column (Pgrandsil-STC-C18, 250 mm×
4.6 mm, 5μm, Beijing, China), a UV detector (Purkinje general,
Beijing, China), and a column oven AT-330 (Autoscience,
Tianjin, China).
1.2 Extraction of capsaicin and dihydrocapsaicin
The ultrasound-assisted extraction of capsaicinoids
from Douban (2.5 g of triturated Douban mixed with 10 mL of
methanol) was carried out in an ultrasonic bath employing
different extraction conditions: power: 200 W; working
frequency: 40 kHz; temperature: 40－80 ℃; solvent: methanol,
ethanol and acetone; extraction time: 5 － 30 min; and extrac-
tion step was repeated 1 － 3 times on each procedure.
After filtration, each filtered solution was filled up to 10
mL with methanol. The extracts were passed through a
0.45μm Millipore nylon filter before HPLC analysis.
1.3 HPLC analysis
HPLC analysis was carried out on a Purkinje liquid
chromatograph model L9 (Purkinje general, Beijing, China).
The mobile phase was a mixture of methanol and water
(75:25, V/V) with a flow rate of 1.0 mL/min. The temperature of
separation column was maintained at 30.0 ℃, the injection
volume was 20μL, and the UV wavelength was 280 nm.
All samples were analyzed in triplicate and where
appropriate, values are reported as mean standards devia-
tion (SD). The standard deviation value was in the range of
5%.
1.4 Calculation
Quantification was performed of the capsaicin and
2012, Vol. 33, No. 04 105
dihydrocapsaicin present in Douban. Capsaicin and
dihydrocapsaicin were quantified from the calibration curves
(y=7575.5x+611.59 for capsaicin (R2=0.9992) and y=7761.2x－
329.58 for dihydrocapsaicin (R2=0.99932) obtained from the
standard solutions. The capsaicinoids content was calcu-
lated as the sum of capsaicin and dihydrocapsaicin by fol-
lowing equation[19]:
W=(Wcapsaicin +Wdihydrocapsaicin)/0.9
All samples were analyzed in triplicate and values are
reported as mean standards deviation (SD).
1.5 Stability of the capsaicinoids
A study was made of the stability of the capsaicinoids
in Douban employing different heating conditions: 2.5 g tritu-
rated Douban was weighed into glass beaker, and then heated
in drying oven, the temperature was controlled at 130 －
200 ℃, the heated time was 2 － 8 min.
2 Results and Analysis
2.1 Solvent
For developing a method of extraction, the selection of
solvent was the first step. The solvents that had been used for
extracting capsaicinoids from samples were methanol[5,20-21],
ethanol[10], acetone[4] and tetrahydrofuran[18-19], et al.
Capsaicin
Dihydrocapsaicin
30 Capsaicinoids
25
Content/(mg/kg)
20
15
10
5
0
Methanol Ethanol Acetone
Solvent
Fig.1 Results obtained employing different solvents in
ultrasound-assisted extraction (n=3)
In this experiment, the solvents that had been studied for
extracting the capsaicinoids from the sample matrix were
methanol, ethanol and acetone. Fig.1 showed the content of
capsaicinoids extracted with different solvents tested. It
c o u l d b e o b s e r v e d t h a t me t h a n o l e x t r a ct e d mo r e
capsaicinoids than ethanol and acetone. Therefore, it was
decided to use methanol as the extraction solvent in this
study.
2.2 Extraction temperature
Temperature affected many physical properties such as
viscosity, diffusivity, solubility, vapor pressure and surface
 106 2012, Vol. 33, No. 04 食品科学 ※工艺技术

tension. However, the main effect of ultrasounds was resulted in a better mass transfer of solute constituents from
cavitational bubbles collapse[13]. The effect of temperature on the plant materials to solvent. The disruption of plant cells by
the release of capsaicinoids extracted by methanol was microjet after the cavitation bubble collapsed could increase
shown in Fig.2. It was found that the increase in temperature the rate of solvent penetration into plant tissue. The release
from 40 to 70 ℃ enhanced the capsaicinoids content. And a rate of capsaicinoids was very high at the beginning due to
significant decrease was obtained when the temperature in- the effect of the capsaicinoids concentration gradient be-
creased to 80 ℃. tween the solvent and the plant material and the ease of
Capsaicin extraction from the outer part of particles in the early period.
Dihydrocapsaicin Afterwards the release rate of capsaicinoids considerably
Capsaicinoids
45 decreased because of the lower concentration gradient and
40
Content/(mg/kg)

35 because the remaining capsaicinoids were located in the in-

30
25 ner part[13]. Therefore, 5 min was selected as the extraction
20
15 time, since this was sufficient to extract most of the
10 capsaicinoids present in samples using less extraction time.
5
0 2.4 Extraction step
40 50 60 70 80
Temperature/℃ The ultrasound-assisted extraction of capsaicinoids
Fig.2 Results obtained employing different temperatures in from Douban was carried out in an ultrasonic bath repeated
ultrasound-assisted extraction (n=3) 1 － 3 times on each procedure.
The first time extraction was carried out as following
2.3 Extraction time conditions: 2.5 g of triturated Douban mixed with 10 mL of
Generally, by increasing the extraction time, the quantity methanol; power: 200W; working frequency: 40 kHz;
of analysis extracted is increased[10]. In this study, extraction temperature: 70 ℃; solvent: methanol; extraction time: 5 min.
times of 5, 10, 15, 20, 25 min and 30 min were evaluated. Seen After filtration, the final volume was adjusted to 10 mL by
from Fig.3, an increase of the content of capsaicinoids was addition of methanol.
obtained with the increase of the extraction time. The extrac- The second and third time of extraction was extracted
tion rate of capsaicinoids was very high during the first 5 from the residue of last time extraction with 10 mL of methanol.
min of the extraction. After that, the content of capsaicinoids Also, each filtrate was filled up to 10 mL with methanol.
gradually increased with the extraction time. The content of Each extracts was analyzed by HPLC respectively. The
capsaicinoids increased 15% when extracted 30 min com- amounts of capsaicinoids were the sum of the individual
pared with 5 min, but it was a waste of time (the extraction extracted capsaicinoids. The results were showed in (Fig.4).
time was increased 500%). In sequential extractions, 99% of the extracted
Capsaicin capsaicinoids was contained in the first extraction filtrate[20].
Dihydrocapsaicin But in this study, 99% of the extracted capsaicinoids was
Capsaicinoids
contained in the second extraction filtrate (Fig.4). So we chose
50
extracted twice as the extraction condition.
Content/(mg/kg)

40
30
20 Capsaicin
50
10 Dihydrocapsaicin
Content/(mg/kg)

40 Capsaicinoids
0
0 5 10 15 20 25 30 30
Time/min 20
Fig.3 Results obtained employing different extraction times in 10
ultrasound-assisted extraction (n=3)
0
1 2 3
UAE significantly improved the extraction yield. Since
Extraction step
ultrasound could accelerate swelling and hydration and Fig.4 Results obtained employing different extraction steps in
caused an enlargement in the pores of the plant cell walls, it ultrasound-assisted extraction (n=3)
※工艺技术
2.5 Stability of capsaicinoids
The capsaicinoids in Douban attribute food of pun-
gency and special flavor when cooking. In heated processing,
the degradation of capsaicinoids will influence the pungency
of Douban. So, we studied the stability of capsaicinoids in
Douban at heating temperatures ranging from 25 to 200 ℃.
70
60
Content/(mg/kg)
50
40
2 min
30
4 min
20 8 min
10
0 capsaicin[J]. Journal of Food Research and Development, 2006, 27(7):
25 130 150 170
Temperature/℃
Fig.5 Effects of different temperatures and heating times on the
stability of capsaicinoids in heated processing (n=3)
Fig.5 presented the average content (n=3) of
capsaicinoids in Douban that heated at temperatures ranging
from 25 to 200 ℃ for a period of 2 to 8 min. Compared with the
sample that heated at 25 ℃, a marked decrease on the con-
tent of capsaicinoids in samples was detected following the
increasing of temperature. The degradation of the
capsaicinoids began to be seen at 130 ℃. Similar results
could be observed in other studies describing degradation
of the capsaicinoids began to be seen at 150 ℃[10,18]. And a
very striking degradation of the capsaicinoids was observed
at 200 ℃. At the same heated temperature, the content of
capsaicinoids decreased following the increasing of heated
time except that at 150 ℃ and 200 ℃.
3 Conclusions
A fast method had been developed for the extraction of
utilizing ultrasound-assisted extraction. In this experiment,
methanol was a better solvent to extract capsaicinoids from
Douban compared with ethanol and acetone. The increased
temperature from 40 up to 70 ℃ enhanced the extraction of
capsaicinoids; however, a significant decrease was observed
when the temperature increased to 80 ℃. An increase of the
content of capsaicinoids was obtained with the increase of
the extraction time. The extraction rate of capsaicinoids was
very high during the first 5 min of the extraction. The suit-
able procedure for extracting capsaicinoids from Douban was
2.5 g of triturated Douban with 10 mL of methanol with a
食品科学 2012, Vol. 33, No. 04 107
controlled temperature of 70 ℃ for a period of 5 min; extrac-
tion step was repeated twice on each procedure. The ultra-
sonic bath was working at a power of 200W and a frequency
of 40 kHz. When heated, the degradation of the capsaicinoids
began to be seen at 130 ℃. And at the same heated
temperature, the content of capsaicinoids decreased follow-
ing the increasing of heated time except that at 150 ℃ and
200 ℃. This would allow us to select a suitable heat
temperature that would have the best flavor without causing
degradation of the capsaicinoids.
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