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Antioxidant activities of several Chinese medicine herbs

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The antioxidant activity (AA) of ethyl acetate extracts of Caesalpinia sappan, Lithospermum erythrorhizon, Anemarrhena asphodeloides, Paris polyphylla and Illicium verum were tested in refined peanut oil at 60 ± 0.5 °C. The concentrations of the extracts added were 0.20% (w/w). The rate of oxidation was assessed by the measurement of peroxide value (PV) and calculation of such characteristics as induction period (IP), when PV reaches 20 meq kg−1, protection factor (PF), which is the ratio of `IP of the sample with additive' and `IP of the sample without additive', and AA (the ratio of `IP increase of the sample with extract' and `IP increase of the sample with butylated hydroxytoluene'). All of C. sappan, L. erythrorhizon extracts and their combinations were found to be high effective in peanut oil. But the extracts of A. asphodeloides, P. polyphylla and I. verum slightly decrease the formation of peroxides in peanut oil as compared with pure oil.
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Antioxidant activities of several Chinese medicine herbs
Pan Yingming
a,*
, Liang Ying
b
, Wang Hengshan
a
, Liang Min
a
a
School of Chemistry and Chemical Engineering, Guangxi Normal University, 15 Yucai Road, Guilin 541004, PR China
b
The Eighth Department, Guilin Institute of Electronic Technology, Guilin 541004, PR China
Received 14 November 2003; received in revised form 9 February 2004; accepted 12 February 2004
Abstract
The antioxidant activity (AA) of ethyl acetate extracts of Caesalpinia sappan,Lithospermum erythrorhizon,Anemarrhena as-
phodeloides,Paris polyphylla and Illicium verum were tested in refined peanut oil at 60 0.5 °C. The concentrations of the extracts
added were 0.20% (w/w). The rate of oxidation was assessed by the measurement of peroxide value (PV) and calculation of such
characteristics as induction period (IP), when PV reaches 20 meq kg1, protection factor (PF), which is the ratio of ÔIP of the sample
with additiveÕand ÔIP of the sample without additiveÕ, and AA (the ratio of ÔIP increase of the sample with extractÕand ÔIP increase of
the sample with butylated hydroxytolueneÕ). All of C. sappan,L. erythrorhizon extracts and their combinations were found to be high
effective in peanut oil. But the extracts of A. asphodeloides,P. polyphylla and I. verum slightly decrease the formation of peroxides in
peanut oil as compared with pure oil.
Ó2004 Elsevier Ltd. All rights reserved.
Keywords: Caesalpinia sappan;Lithospermum erythrorhizon;Anemarrhena asphodeloides;Paris polyphylla;Illicium verum; Extracts; Antioxidant
activity; Peanut oil
1. Introduction
Governmental medical authorities and consumers are
concerned about the safety of their food and about the
potential effect of synthetic additives on their health.
During the last few decades an intensive testing of the
safety of synthetic food additives has been carried out
and many of them have been found to possess some
toxic activity (Bandoniene, Pukalskas, Venskutonis, &
Gruzdiene, 2000). As a result, search of natural substi-
tutes which in most cases are considered as generally
recognised as safe (GRAS) substances has increased
considerably.
Such a tendency can also be applied to synthetic an-
tioxidants, such us butylated hydroxytoluene (BHT),
butylated hydroxyanisole (BHA), etc. BHA has been
shown to cause lesion formation in the rat forestomach.
Moreover, several studies have shown that BHT may
cause internal and external haemorrhaging at high doses
that is severe enough to cause death in some strains of
mice and guinea pig (Shahidi & Wanasundara, 1992).
Accordingly, there is a strong argument for the effective
isolation of organic antioxidants from natural sources as
alternatives to prevent deterioration of foods (Kikuzaki
& Nakatani, 1993). The number of reports about iso-
lation and testing of natural, mainly of plant origin,
antioxidants has increased during the last twenty im-
mensely(Mallet, Cerrati, Ucciani, Gamisons, & Gruber,
1994; Scartezzini & Speroni, 2000; Xiong, Yang, Zhang,
& Xiao, 2001). These attempts have led to the devel-
opment of very effective natural antioxidants from
rosemary (Rosmarimus officinalis) and sage (Salvia offi-
cinalis), which are now available commercially and are
safe in food (Bishov, Masuoka, & Kapsalis, 1977;
Djarmati, Jankov, Schwirtlich, Djulinac, & Djordjevic,
1991; Pokorny, Nquyen, & Korczak, 1997).
A great number of different spices and aromatic herbs
have been tested for their antioxidant activity (AA),
however. there are still many plants, which were not
examined on this matter or the knowledge about their
antioxidative properties are very rarely. Caesalpinia
sappan,Lithospermum erythrorhizon,Anemarrhena
*
Corresponding author. Tel.: +86-773-584-6279; fax: +86-773-581-
2383.
E-mail address: panym2004@yahoo.com.cn (P. Yingming).
0308-8146/$ - see front matter Ó2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2004.02.002
Food Chemistry 88 (2004) 347–350
www.elsevier.com/locate/foodchem
Food
Chemistry
asphodeloides,Paris polyphylla and Illicium verum are
among such plants. These plants were investigated from
some other points of view, mostly regarding their me-
dicinal properties, pigment, essential oil and flavonoid
composition(Fukui, Feroj Hasan, Ueoka, & Kyo, 1998;
Niranjan Reddy, Ravikanth, Jansi Lakshmi, Suryan-
arayan Murty, & Venkateswarlu, 2003; Sy & Brown,
1998; Tuan & Ilangantileke, 1997; Zhang et al., 1999;
Zhou, Yang, Li, Wang, & Wu, 2003).
The present study was undertaken to perform the
screening of antioxidant properties of several Chinese
medicine herbs, C. sappan,L. erythrorhizon,A. asphod-
eloides,P. polyphylla and I. verum. For this purpose
ethyl acetate extracts obtained from these plants were
added to the peanut oil and oxidative deterioration
(formation of peroxides) of it was measured at different
time periods during storage in an oven at 60 0:5°C.
BHT established strong antioxidant effects, which were
used for comparison reasons.
2. Materials and methods
2.1. Materials
The following reagents were used: synthetic antioxi-
dant 2,6-di-tert-butyl-4-methylphenol (BHT) (C.P.,
Shanghai Huayuan Fine Chemical Industry CO.,LTD,
China), ethyl acetate (A.R., China National Medicine
Group, China), chloroform (A.R., China National
Medicine Group, China), acetic acid (A.R., pure, Gu-
angzhou Chemical Reagent Factory, China), potassium
iodide (A.R., China National Medicine Group, China),
sodium thiosulfate (A.R., China National Medicine
Group, China), starch soluble (A.R., China National
Medicine Group, China), potassium dichromate (A.R.,
Guangzhou Chemical Reagent Factory, China), sulfuric
acid (A.R., 98%, China National Medicine Group,
China).
C. sappan,L. erythrorhizon,A. asphodeloides,P. po-
lyphylla and I. verum were obtained from Guilin Phar-
maceuticals Group of China.
The fresh peanut oil was bought from Guilin country
market, and deeply refined. It contained no synthetic
antioxidants (acid value 3.2 mg KOH g1, linoleic acid
6% and peroxide value (PV) 0.85 meq kg1).
2.2. Methods
2.2.1. Preparation of plant extracts
The herbs were ground (max particle size 0.4 mm)
and 50 g of comminuted material extracted with 500 mL
ethyl acetate (A.R., China) in a Soxhlet apparatus dur-
ing 24 h. Solvent was vaporated in a RE-52AA rotav-
apour (Shanghai Yarong Biochemistry Instrument
Factory, China) by using a water bath (60 °C) and a
SHB-bA water-circulation multifunction vacuum pump
(Zhengzhou Great Wall Scientific Industry and Trade
CO., LTD, China). The extracts were finally dried in a
DZF-1B vacuum drier (Shanghai Yuejin Medical In-
strument CO., LTD, China) at 30 °C and 0.07 MPa. Dry
extracts were stored in a freezer until use. The yields of
the plant extracts were as follows: C. sappan (CE) –
10.32%, L. erythrorhizon (LE) – 13.42%, A. asphodelo-
ides (AE) – 16.57%, P. polyphylla (PE) – 11.70%, I. ve-
rum (IE) – 12.89%.
Then, they were mixed to form combinations of an-
tioxidants, L1C1E(MLE:MCE ¼1:1), L2C1E(MLE:MCE ¼
2:1), L3C1E(MLE:MCE ¼3:1), L1C2E(MLE:MCE ¼1:2),
L1C3E(MLE:MCE ¼1:3), L1C1A1E(MLE:MCE :MAE ¼1:
1:1), L2C1A1E(MLE:MCE :MAE ¼2:1:1), L3C1A1E(M
LE:
MCE:MAE ¼3:1:1), L1C2A1E(MLE :MCE :MAE ¼1:2:1),
L1C3A1E(MLE:MCE :MAE ¼1:3:1).
2.2.2. Introduction of extracts into the oil
Calculated amounts of the extracts (0.2% of the oil
weight) were added to the 50 g peanut oil. The additive
was mixed into the oil with a magnetic stirrer. Synthetic
antioxidant BHT were used as reference substances for
comparative purposes.
2.2.3. Methods of assessment of oil oxidation and stability
The oil samples (50 g each) were placed in open 100
mL volume beakers. The oxidative deterioration of
samples was studied by Schaal oven test as described by
Economou, Oreopoulou, and Thomopoulos (1991). The
experiments were repeated twice. When the differences
between the replicates were rather big, then the mea-
surements were repeated. However, such cases were ex-
ceptionally rare. In all cases standard deviation was in
the range of 2–8% from the mean. A blank sample was
prepared under the same conditions, without adding any
additives. The rate of autoxidation of peanut oil was
estimated according to the increase of its PV, which was
determined by the method as described by Bandoniene
et al. (2000).
The changes of the induction period (IP) of oil after
the addition of each extract, was determined as a func-
tion of its concentration in oil. The IP was considered as
the number of hours needed for the PV of the sample to
reach the value of 20 meq kg1(Wanasundara &
Shahidi, 1994). Protection factor (PF) values of peanut
oil and antioxidant activities (AA) of the extracts were
calculated by the following formulas:
PF ¼IPX
IPK
;
AA ¼IPXIPK
IPBHT IPK
;
where: IPX– induction period of sample with additive,
h; IPK– induction period of sample without additive, h;
348 P. Yingming et al. / Food Chemistry 88 (2004) 347–350
IPBHT – induction period of sample with added synthetic
antioxidant BHT, h. The following scale is proposed for
the PF values: 1.0–1.5 (very low), 1.5–2.0 (low), 2.0–2.5
(medium), 2.5–3.0 (high), >3.0 (very high) (Ahmad,
Hakim, & Shehata, 1983). Actually, PF is defined as a
stability value with additive divided by that of the blank
sample.
3. Results and discussion
The data for peanut oil autoxidation, measured as a
changes of PV, at 60 0:5°C after addition of extracts
of C. sappan,L. erythrorhizon,A. asphodeloides,P. po-
lyphylla,I. verum and their combinations are presented
in Table 1. The concentrations of the extracts in oil,
calculated on a dry weight basis, is 0.20% (w/w). It is
evident that all extracts and their combinations in gen-
eral showed some oil stabilising effect.
The extracts obtained from C. sappan,Lithospermum
erythrorhizon and their combinations were found to be
the most effective natural antioxidants. The effect of C.
sappan and L. erythrorhizon extracts and some of their
combinations on the stability of peanut oil during ac-
celerated oxidation storage conditions was better than
the effect of butylated hydroxytoluene (BHT) at the
same concentration. The most important finding of this
study was the strong activity of C. sappan and Litho-
spermum erythrorhizon extracts which was according to
our knowledge revealed for the first time. For instance,
PV of peanut oil with 0.20% of CE and LE after 20
days of storage was 22.50–24.10 meq kg1, whereas in
blank samples it increased to 114.78 meq kg1only
after 10 day of storage, in the samples with the extracts
from other herbs to 102–108 meq kg1. Having in mind
that BHT is a pure compound while the extracts are
complex mixtures containing ineffective substances in
terms of their antioxidative activity or even some
amounts of pro-oxidative compounds, it could be sug-
gested that C. sappan and L. erythrorhizon contains
very strong constituents retarding lipid peroxidation.
Therefore, the structures of these constituents in C.
sappan and L. erythrorhizon are target for further
investigations.
The relative antioxidant efficiencies of C. sappan, L.
erythrorhizon,A. asphodeloides,P. polyphylla and I. ve-
rum and their combinations are compared in Fig. 1
where IP of peanut oil is presented after the addition of
extracts in oil.
Data provided in Fig. 1 also show that C. sappan,L.
erythrorhizon extracts and their combinations are much
more effective in stabilizing peanut oil than other extracts
used in this experiment. The effectiveness of the other
plant extracts decreased followed this order: Anemarrhena
asphodeloides >Paris polyphylla >Illicium verum at a
concentration of 0.20%.
PFs and AAs of the extracts are presented in Table 2.
The effectiveness of antioxidants was compared accord-
ing to their stability values and PFs. The effectiveness of
antioxidants under the conditions used is ranged in the
following descending order: L2C1A1E>L
1C1A1E>L
3C1
A1E>L
1C2A1E>L
3C1E>LE>L
1C3E>CE>L
2C1E>
BHT>L
1C3A1E>L
1C2E>L
1C1E>AE>IE>PE>blank
sample.
C. sappan,L. erythrorhizon extracts and their com-
binations exhibited a ‘‘very high’’ AA (PF > 3), A. as-
phodeloides,P. polyphylla,I. verum exhibited ‘‘very low’’
AA (PF of 1–1.5). The structures of isolated constituents
Table 1
Effect of various extracts and their combinations (0.20%) on the formation of peroxides in peanut oil at 60 0:5°C
Additive PVs (meq kg1) after different storage time (days)
0124681012141720
Blank 0.92 2.10 3.82 21.2 54.61 84.46 114.78
BHT 0.92 2.29 2.88 4.34 6.08 8.07 11.13 15.06 19.50 22.88 26.52
LE 0.92 1.98 2.25 3.28 5.66 8.96 10.35 14.00 16.83 18.96 22.58
CE 0.92 2.04 2.76 4.33 6.24 8.45 11.42 14.23 16.46 19.98 24.08
AE 0.92 2.30 2.66 13.00 27.50 45.75 55.49 64.35 79.87 91.62 107.55
PE 0.92 2.51 3.13 14.23 29.65 46.22 58.67 72.44 81.65 90.12 103.50
IE 0.92 2.63 4.65 15.14 29.23 48.22 52.01 64.55 77.3 88.97 102.97
L1C1E 0.92 1.58 1.87 3.00 4.69 8.28 10.62 17.67 21.92 33.46 43.67
L2C1E 0.92 2.03 2.25 3.05 4.28 7.23 9.59 14.05 18.66 21.28 28.55
L3C1E 0.92 1.55 1.81 3.06 4.15 6.37 9.00 12.64 15.22 18.36 21.05
L1C2E 0.92 1.97 2.06 3.41 4.71 9.29 11.03 15.17 21.97 31.94 38.63
L1C3E 0.92 1.64 2.04 3.46 5.62 9.80 12.40 14.31 17.84 20.05 23.59
L1C1A1E 0.92 1.7 1.93 2.94 4.3 6.91 9.84 11.76 13.50 16.74 20.26
L2C1A1E 0.92 1.87 2.04 3.51 4.38 6.03 8.64 10.34 12.02 15.33 18.65
L3C1A1E 0.92 1.76 2.05 3.29 4.13 6.78 9.31 11.16 15.31 17.88 20.62
L1C2A1E 0.92 1.71 1.80 3.19 4.94 8.54 11.95 13.99 16.14 18.15 20.83
L1C3A1E 0.92 1.94 2.10 2.83 5.35 10.79 13.22 16.81 20.35 23.65 27.54
P. Yingming et al. / Food Chemistry 88 (2004) 347–350 349
need to be elucidated and assessed in order to obtain
more precise results.
4. Conclusion
The results of this study suggest that C. sappan,L.
erythrorhizon extracts and some their combinations were
more efficient than BHT at a similar concentration in
peanut oil at 60 0:5°C.
Strong AA of C. sappan,L. erythrorhizon extracts has
been reported for the first time, which gives a strong
impact for expanding the investigations of constituents
responsible for the protection of oil against oxidation.
Acknowledgements
This study was supported by the ’’foundation for
scholars come back from abroad’’ of Guangxi province
(No. 0009007).
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Table 2
AA of extracts (0.20%) and their effect on the stability of peanut oil
Additive PFaAAb
Without additive 1.00
BHT 3.75 1.00
LE 4.80 1.38
CE 4.53 1.28
AE 1.40 0.51
PE 1.23 0.08
IE 1.33 0.12
L1C1E 3.53 0.92
L2C1E 4.27 1.19
L3C1E 5.07 1.48
L1C2E 3.67 0.97
L1C3E 4.56 1.29
L1C1A1E 5.31 1.56
L2C1A1E 5.80 1.74
L3C1A1E 5.27 1.55
L1C2A1E 5.24 1.54
L1C3A1E 3.69 0.98
a
PF is the ratio of IP of the sample with additive with IP of the
sample without additive.
b
AA was calculated in comparison with BHT at the concentration
0.20%.
0
5
10
15
20
25
IP(days)
BHT LE CE AE PE IE
L1C1E L2C1E L3C1E L1C2E L1C3E L1C1A1E
L2C1A1E L3C1A1E L1C2A1E L1C3A1E
Fig. 1. Changes of the IP of peanut oil after the addition extracts and
BHT at concentrations 0.20%.
350 P. Yingming et al. / Food Chemistry 88 (2004) 347–350
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Oxidative stress overwhelms the antioxidant mechanisms of living systems, with active involvement in the pathogenesis of several diseases. Natives of Gangnim in the Plateau State of Nigeria may be unknowingly endowed with some protective advantages against oxidative stress for their habitual consumption of Artemisia annua tea. The antioxidant activities of A. annua extracts were determined using in vitro methods and the inhibitory potentials of twenty-nine (29) bioactive compounds of the plant against oxidative stress target proteins were assessed through molecular docking analysis. These extracts showed significantly high activities in scavenging nitric oxide, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing ferric (Fe3+) to ferrous (Fe2+) iron. Virtually, none of the bioactive compounds binds to the active site of the antioxidant protein targets. Rather, 72.41, 93.10 and 75.86% of these compounds bind with high binding affinity to the activator binding sites of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) respectively. 7,8-dimethylalloxazine (-8.10 kcal/mol) ranked highest as a prospective inhibitor of xanthine oxidase (XOX). The antioxidant activity exhibited by the extracts of the locally cultivated A. annua and the molecular interactions of its bioactive compounds against the protein targets used predict that oxidative stress inhibition could be effectively achieved with these phytochemicals.
... The above result correlates with Thangavel et al. and Pratima et al. [31,32] . Phenolic content of the plant is directly proportional to the antioxidant activity [33] as it absorbs the free radicals by decomposing peroxide [34,35] . Our results confirmed the above said statement. ...
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This review gives, for the first time, a systematic presentation and discussion on the chemistry and use of brazilein in foods. Processes of isolation, purification and quantification of this alternative pigment are firstly reviewed. Molecular structure and color stabilities as well as ways to enhance stability of the pigment are then discussed. Selected applications of the pigment in foods are given. Based on the review of the literature, future studies should focus on the isolation and purification of the pigment prior to its use in foods. Extraction yield and purity of brazilein obtained from the different methods should also be compared. Since the pigment is very sensitive to pH change, its stability should be enhanced prior to its use. Co-pigmentation is among the methods that exhibits potential for stability enhancement of the pigment.
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Investigation of natural antioxidant and synergistic activity of some food ingredients included twenty spices, herbs and plant protein hydrolyzates. Of the spices tested, clove, cinnamon, sage, rosemary, mace, oregano, allspice and nutmeg were highly antioxidant and when coupled with BHA, these spices acted as strong synergists. Autolyzed yeast proteins (AYP), when combined with some of these spices, significantly extended the stability of the model food system. Results of tests of freeze-dried model systems consisting of corn oil and carboxymethyl cellulose (CMC) stored at 65°C suggest possible applications for stabilizing oxygen sensitive foods against quality deterioration.
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