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Quenching activities of common hydrophilic and lipophilic antioxidants against singlet oxygen using chemiluminescence detection system

  • Fuji Chemical Industries Co. Ltd.

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The singlet oxygen quenching activities among common hydrophilic and lipophilic antioxidants such as polyphenols, tocopherols, carotenoids, ascorbic acid, coenzyme Q10 and α-lipoic acid were recorded under the same test condition: the chemiluminescence detection system for direct 1O2 counting using the thermodissociable endoperoxides of 1,4-dimethylnaphthalene as 1O2 generator in DMF : CDCl3 (9 : 1). Carotenoids exhibited larger total quenching rate constants than other antioxidants, with astaxanthin showing the strongest activity. α-Tocopherol and α-lipoic acid showed considerable activities, whereas the activities of ascorbic acid, CoQ10 and polyphenols were only slight; these included capsaicin, probucol, edaravon, BHT and Trolox. This system has the potential of being a powerful tool to evaluate the quenching activity against singlet oxygen for various hydrophilic and lipophilic compounds.
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VOLUME 11 (2007)
Quenching Activities of Common Hydrophilic and Lipophilic Antioxidants
against Singlet Oxygen Using Chemiluminescence Detection System
Yasuhiro Nishida*, Eiji Yamashita and Wataru Miki
Institute for Food Science Research, Fuji Chemical Industry CO., Ltd., 55 Yokohoonji, Kamiichi, Toyama 930-0397,
The singlet oxygen quenching activities among common hydrophilic and lipophilic antioxidants such as
polyphenols, tocopherols, carotenoids, ascorbic acid, coenzyme Q10 and α-lipoic acid were recorded under the
same test condition: the chemiluminescence detection system for direct 1O2 counting using the
thermodissociable endoperoxides of 1,4-dimethylnaphthalene as 1O2 generator in DMF : CDCl3 (9 : 1).
Carotenoids exhibited larger total quenching rate constants than other antioxidants, with astaxanthin showing
the strongest activity. α-Tocopherol and α-lipoic acid showed considerable activities, whereas the activities of
ascorbic acid, CoQ10 and polyphenols were only slight; these included capsaicin, probucol, edaravon, BHT
and Trolox. This system has the potential of being a powerful tool to evaluate the quenching activity against
singlet oxygen for various hydrophilic and lipophilic compounds.
1. Introduction
Living organisms possess defense mechanisms
against oxidative damage. One of the most important
ways is using an antioxidants, such as ascorbic acid,
polyphenols, coenzyme Q10 (CoQ10), tocopherols or
carotenoids [2], for quenching and/or scavenging
against reactive oxygen species (ROS).
Singlet oxygen (1O2) is a non-radical ROS with
one of the strongest activities. It directly damages onto
biological lipids, proteins and DNA, which are related
to serious diseases such as diabetes, hypertension and
cancer [1,2]. It would be valuable to search an effective
quencher against 1O2 and to develop its methodology.
Di Mascio [3] reported that lycopene showed the
highest activity among carotenoids and tocopherols by
Germanium photodiode detection system in EtOH :
CHCl3 : H2O (50 : 50 : 1) using the thermodissociable
endoperoxides of a naphthalene derivative (NDPO2) as
a 1O2 generator. One of the authors [4] evaluated the
activities of marine carotenoids and α-tocopherol in
two solvent systems, CDCl3 and CDCl3 : CD3OD (2 :
1) by the chemiluminescence detection system for
direct 1O2 counting using the thermodissociable
endoperoxides of 1,4-dimethylnaphthalene (EDN) as
1O2 generator. And it was found that astaxanthin
showed the strongest activity.
The activities of carotenoids and tocopherols were
revealed by both studies, whereas those of the other
compound groups remain largely unknown. We
therefore wanted to compare the activities of common
antioxidants in nature such as ascorbic acid,
polyphenols, α-lipoic acid, CoQ10 and others to those
of carotenoids and tocopherols. Here we report the
direct comparison of the quenching activities against
1O2 among the antioxidants with not only lipophilic but
hydrophilic property under the same test conditions.
*Corresponding author. E-mail:
BHT, butyleted hydroxytoluene; CoQ10, coenzyme Q10; EDN, endoperoxides of 1,4-dimethyl naphthalene;
EGCG, epigallocatechin gallate; LDL, low-density lipoprotein; NDPO2, naphthalene-1,4-dipropionate
endoperoxide; 1O2, singlet oxygen; QOL, quality of life; ROS, reactive oxygen species.
Fig. 1 Chemical structures of tested compounds
2. Experimental
2.1. Test compounds.
Astaxanthin, lutein, α-lipoic acid, ubiquinone-10
(CoQ10), caffeic acid, quercetin, resveratrol, gallic
acid, pyrocatechol, pyrogallol, BHT and sesamin were
purchased from Sigma-Aldrich (St. Louis, MO, USA),
L(+)-ascorbic acid, α-tocopherol, probucol,
canthaxanthin and lycopene from Wako Pure Chemical
Industries, Ltd. (Osaka, Japan ), β-cryptoxanthin from
Extrasynthase (Genay, France), Trolox and edaravon
(MCI-186) from Cosmo Bio Co., Ltd. (Tokyo, Japan)
and curcumin I, (-)-epigallocatechin gallate (EGCG)
and capsaicin from Nagara Science Co., Ltd. (Gifu,
Japan). β-Carotene was a gift from Prof. H. Hashimoto
of Osaka City University. Fucoxanthin was extracted
from the brown algae Undaria pinnatifida and
Laminaria japonica. Recrystallization and/or
chromatography of all these compounds resulted in
obtaining a purity greater than 99%.
2.2. Measurement of 1O2 quenching activity
As one of the authors previously reported [4],
thermodissociable EDN prepared from
1,4-dimethylnapthalene (purchased from
Sigma-Aldrich in St. Louis, MO, USA) was used as 1O2
generator. EDN was dissolved with CDCl3 and stored
at below 0 ºC until used. It could release molecular
oxygen in the singlet state (1Δg) at 37 ºC.
Chemiluminescence emissions from 1O2 were counted
with a chemiluminescence detector, AccuFlex Lumi
400 (Aloka, Japan).
One hundred eighty micro litters of CDCl3 or a
mixture of CDCl3 : CD3OD (2:1) or DMF containing
0.01 to 50,000 μM of each compound was placed in a
thermostated glass tube (12φ X 75 mm) at 37 ºC.
Chemiluminescence counting was started just after
addition of the EDN in CDCl3 at the final concentration
of 50 mM, and was counted for 60 seconds. Both
chemiluminescence counts of a control (S0) without
any test compound, and a sample (S) with the identical
test compound were recorded. The total quenching
constant, generally for total quenching by chemical
reaction and/or physical quenching, kT = kq + kr, was
analyzed on a Stern-Volmer plot, which is based on the
following equation [5],
S0/S = 1+ kTkd
-1[Q] (1)
where kq is the physical quenching rate constant, kr is
the chemical reaction rate constant, kd is the first-order
decay rate constant of singlet oxygen in the solvent,
and [Q] is concentration of the test compounds. Total
quenching constant kT was used to evaluate the activity
of the each test compound.
3. Results and Discussion
Within the range of the concentrations actually
tested each antioxidant was dissolved in the solvent at
37 ºC.
Fig. 2 showed the Stern-Volmer plots of
astaxanthin, β-carotene, lycopene, α-tocopherol and
α-lipoic acid in CDCl3, CDCl3 : CD3OD (2 : 1) and
DMF : CDCl3 (9 : 1). A high linearity meaning more
than 0.9 of r2 value was observed in the each plot. The
plots of other test compounds were similar to those
(data not shown).
Total quenching constant (kT = kq + kr) of each test
compound is shown in Table 1. The activities of
canthaxanthin (in CDCl3), α-carotene, β-cryptoxanthin,
fucoxanthin (in CDCl3), lycopene, lutein (in CDCl3 :
CD3OD (2 : 1)), α-tocopherol (in CDCl3 : CD3OD (2 :
1)), CoQ10 and α-lipoic acid were additionally
recorded by the same method as the former study [4]
and a similar tendency was observed. Briefly,
carotenoids showed stronger 1O2 quenching activities
than α-tocopherol as well as CoQ10 and α-lipoic acid
which are recognized as common antioxidants.
In the case of carotenoids, a number of conjugated
double bonds including C=C and C=O were found to
contribute the quenching activity. In CDCl3 represented
as the lipophilic system, lycopene showed the largest
value. And in CDCl3 : CD3OD (2 : 1) with more
hydrophilicity, astaxanthin did so.
Both hydrophilic and lipophilic common
antioxidants were directly compared in the new system
using DMF : CDCl3 (9 : 1). All carotenoids exhibited
larger kT value than other antioxidants. Moreover,
astaxanthin showed the strongest activity among
carotenoids tested. The hydroxyl groups in the
carotenoid molecule were found to contribute slightly
to the activity in the solvent, while the carbonyl groups
were also found in CDCl3 : CD3OD (2 : 1). The values
of α-tocopherol and α-lipoic acid were relatively large.
Ascorbic acid, CoQ10 and polyphenols such as EGCG
represented as catechins, quercetin as flavonoids,
curcumin as curcumnoids, resveratrol as stilbenoids,
gallic acid as tannins, sesamin as lignan, pyrocatechol,
caffeic acid and pyrogallol had weaker activities.
Weaker activities were also noted in capsaicin,
probucol and edaravon as medicines, BHT used as an
antioxidant food additive and Trolox
(6-hydroxy-2,5,7,8-tetramethyl- chroman-2-carboxylic
acid) which is a reference substance for ORAC
(Oxygen Radical Absorbance Capacity) value. They
might rather be singlet oxygen quenchers than free
radical scavengers against superoxide anion or
hydroxyl radicals. This system has the potential of
being a powerful tool to evaluate the quenching activity
against singlet oxygen for various hydrophilic and
lipophilic compounds.
Overall, astaxanthin exhibited the most potent
singlet oxygen quenching activity among the
compounds tested in this study because it showed a
stable superior property under the three different
conditions. Astaxanthin is widely distributed in fish
and shellfish, crustaceans, zoo- and phyto-planktons,
bacteria and so on, particularly in marine organisms. In
fact, the first isolation and identification was
accomplished in 1938 from the lobster, Astacus
gammarus [6], and numerous studies were carried out
over a long period. It is reported that the biological
activity of astaxanthin originated from potent 1O2
quenching and lipid peroxidation suppressing activities
[7]. Various human benefits for human health have
been recognized to date: immunomodulation [8],
anti-stress [9], anti-inflammation [10], LDL cholesterol
oxidation suppression [11], enhanced skin health [12],
improved semen quality [13], attenuation of eye fatigue
[14], sports performance and endurance enhancement
[15], limitations on exercised induced muscle damage
[16], limitations of diabetic nephropathy [17],
improvement of hypertension [18] and metabolic
syndrome [19]. Astaxanthin obviously plays an
important role in promoting QOL to prevent diseases
and maintain a healthy life.
Further study is needed for the evaluation of lipid
peroxidation suppressing activity among common
hydrophilic and lipophilic antioxidants based on this
Fig.2 Stern-Volmer plots of some tested compounds
4. Acknowledgments
We thank Dr. H. Hashimoto of Osaka City
University for supplying β-carotene. We are also
indebted to Dr. V. Wood and Ms. A. Miyashita of Fuji
Chemical Industry Co. Ltd. for valuable discussion.
5. References
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Azumane, A., Kaneko, K., and Yamaguchi, M., J.
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Table 1. Total singlet oxygen quenching rate constants
- Carotene
β- Carotene
β- Cryptoxanthin
Lutei n
L - Ascorbi c a cid
- Toc o p h er o l
- Lipoic acid
Ubiquinone- 10
( - ) - Epigallocatechin gallate
Gallic aci
Compound Tested Concentration
0.01- 15
0.01- 15
0.01- 15
0.01- 15
0.01- 15
0.01- 15
0.01- 15
0.01- 15
0.01- 15
20- 50,000
10- 20,000
10- 10,000
10- 3,000
10- 10,000
10- 10,000
10- 10,000
10- 6,000
10- 10,000
10- 10,000
10- 10,000
10- 10,000
10- 10,000
10- 5,000
10- 10,000
10- 10,000
10- 10,000
10- 20,000
OD (2:1) DMF/CDCl
... AX has antioxidant activity, a well-known characteristic of carotenoids. Aside from its ability to quench a number of reactive oxygen species (ROS) and reactive nitrogen species (RNS), and other free radicals, AX stands out among carotenoids due to its particularly strong singlet oxygen quenching activity [14][15][16]. AX is also well-known for strongly inhibiting the accumulation of lipid peroxides resulting from lipid peroxidation chain reactions [17,18]. In biological environments, AX has been detected in lipid droplets [19], cell membranes [20], or bound to proteins [18,[21][22][23], due to its highly lipophilic properties. ...
... The increase in fat oxidation at low intensity after ET was greater in AX (placebo 0. 23 ± 0.15 g vs. AX 0.76 ± 0.18 g), and was associated with reduced carbohydrate oxidation and improved exercise efficiency in men, but not in women. 14.4% (± 6.2%, p < 0.02), tibialis anterior muscle size (cross-sectional area, CSA) by 2.7% (± 1.0%, p < 0.01), and specific impulse increased by 11.6% (MVC/CSA, ± 6.0%, p = 0.05), respectively, whereas placebo treatment did not alter these characteristics (MVC, 2.9% ± 5.6%; CSA, 0.6% ± 1.2%; MVC/CSA, 2.4 ± 5.7%; all p > 0.6). ...
... * In addition to AX, other nutrients such as antioxidants were used in the study.Nutrients 2022,14,107 ...
Full-text available
Astaxanthin is a member of the carotenoid family that is found abundantly in marine organisms, and has been gaining attention in recent years due to its varied biological/physiological activities. It has been reported that astaxanthin functions both as a pigment, and as an antioxidant with superior free radical quenching capacity. We recently reported that astaxanthin modulated mitochondrial functions by a novel mechanism independent of its antioxidant function. In this paper, we review astaxanthin’s well-known antioxidant activity, and expand on astaxanthin’s lesser-known molecular targets, and its role in mitochondrial energy metabolism.
... The proposed mechanisms of astaxanthin antioxidant action are electron donation, bonding with free radicals to form less active forms and nonreactive products, transportation of radicals along its own carbon chain outside the cell, inhibition of ROS formation, and metal chelation by two adjacent oxygen atoms on the cyclohexene ring. All of these antioxidant activities have been reported by numerous in vitro and in vivo studies [6,8,23,45,51,[86][87][88][89]. ...
... Astaxanthin reduced the release of interleukins, cycloxygenase-2 and nitric oxide generation, and DNA damage induced by radioactivity as well as improved inflammatory-related pathways [110,[128][129][130][131][132][133][134]. It has been found to impart a healthy state by reducing oxidative stress, neutralizing singlet oxygen, scavenging free radicals, inhibiting lipid peroxidation, and improving immunity and muscle health [5,7,89,[135][136][137]. Several studies have also indicated the ability of astaxanthin from shrimp to reduce obesity, hypertension, hyperlipidemia, and heart-related ailments [138][139][140][141]. ...
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In recent years, the food, pharma, and cosmetic industries have shown considerable interest in bioactive molecules of marine origin that show high potential for application as nutraceuticals and therapeutic agents. Astaxanthin, a lipid-soluble and orange-reddish-colored carotenoid pigment, is one of the most investigated pigments. Natural astaxanthin is mainly produced from microalgae, and it shows much stronger antioxidant properties than its synthetic counterpart. This paper aims to summarize and discuss the important aspects and recent findings associated with the possible use of crustacean byproducts as a source of astaxanthin. In the last five years of research on the crustaceans and their byproducts as a source of natural astaxanthin, there are many new findings regarding the astaxanthin content in different species and new green extraction protocols for its extraction. However, there is a lack of information on the amounts of astaxanthin currently obtained from the byproducts as well as on the cost-effectiveness of the astaxanthin production from the byproducts. Improvement in these areas would most certainly contribute to the reduction of waste and reuse in the crustacean processing industry. Successful exploitation of byproducts for recovery of this valuable compound would have both environmental and social benefits. Finally, astaxanthin's strong biological activity and prominent health benefits have been discussed in the paper.
... (ii) Astaxanthin is one of the most powerful antioxidant molecules and can scavenge ROS and could act as a barrier that prevents lipids, pigments, and photosynthetic complexes oxidation. Astaxanthin showed an antioxidant activity against singlet oxygen far higher than β-carotene [23,62]. According to this hypothesis, singlet oxygen was detected in far higher amount in UVM4 compared to the strain that accumulates astaxanthin (Fig. 4C). ...
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Background Astaxanthin is a highly valuable ketocarotenoid with strong antioxidative activity and is natively accumulated upon environmental stress exposure in selected microorganisms. Green microalgae are photosynthetic, unicellular organisms cultivated in artificial systems to produce biomass and industrially relevant bioproducts. While light is required for photosynthesis, fueling carbon fixation processes, application of high irradiance causes photoinhibition and limits biomass productivity. Results Here, we demonstrate that engineered astaxanthin accumulation in the green alga Chlamydomonas reinhardtii conferred high light tolerance, reduced photoinhibition and improved biomass productivity at high irradiances, likely due to strong antioxidant properties of constitutively accumulating astaxanthin. In competitive co-cultivation experiments, astaxanthin-rich Chlamydomonas reinhardtii outcompeted its corresponding parental background strain and even the fast-growing green alga Chlorella vulgaris . Conclusions Metabolic engineering inducing astaxanthin and ketocarotenoids accumulation caused improved high light tolerance and increased biomass productivity in the model species for microalgae Chlamydomonas reinhardtii . Thus, engineering microalgal pigment composition represents a powerful strategy to improve biomass productivities in customized photobioreactors setups. Moreover, engineered astaxanthin accumulation in selected strains could be proposed as a novel strategy to outperform growth of other competing microalgal strains.
... AST is well recognized for its antioxidant potencies. It is 110 times potent antioxidant than vitamin E, 560 times than green tea catechins, 800 times than Co-Q 10, 3000 times than resveratrol and 6000 times than vitamin C [11][12][13]. It is strongly supposed as a completely unique antioxidant and a potent bioactive compound as it shows three concurrent novel peculiarities i.e., powerful anti-oxidation property, safe for human use and acquires best position within the cell membrane [13]. Structurally, AST is present in trans and cis (E and Z) isomeric forms (Fig. 2). ...
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... Miki [6] examined the quenching or scavenging effects of animal carotenoids against active oxygen species, singlet oxygen and hydroxy-radical, and against organic free radicals and reported that the potencies of astaxanthin are approximately 10 times stronger than those of other carotenoids (zeaxanthin, lutein, tunaxanthin, canthaxanthin and beta-carotene) and 100 times greater than those of alpha tocopherol. Nishida et al. [7] reported that astaxanthin exhibited the most potent singlet oxygen-quenching activity among the compounds tested (lutein, α-lipoic acid, ubiquinone-10(CoQ10), caffeic acid, quercetin, resveratrol, gallic acid, pyrocatechol, pyrogallol, BHT, sesamin, L(+)-ascorbic acid, α-tocopherol, probucol, canthaxanthin, lycopene, β-cryptoxanthin, Trolox, edaravon, curcumin, epigallocatechin gallate, capsaicin, β-carotene and fucoxanthin). ASTX inhibits lipid peroxidation 100 to 500-fold more strongly than vitamin E in vitro [8] and has a several-fold greater free radical antioxidant potency than vitamin E and β-carotene [9]. ...
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... As a result, the hypoxia-induced accumulation of M1 macrophages leads to synergistic cytotoxicity with the hypoxic response, and the DAMPs generated by the cytotoxicity further accumulate M1 macrophages, creating a vicious cycle including ROS-mediated oxidative stress. From these viewpoints, AX has a remarkable inhibitory effect on the progression of the fat peroxidation chain reaction and on the direct peroxidation of lipids by singlet oxygen [42,43]. It has also been reported that AX inhibits the activation of the classical NFκB pathway through various cytokines and NO stimulation in the strain of monocytes, RAW264.7, and thereby suppresses excessive inflammatory responses [44]. ...
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Recently, obesity-induced insulin resistance, type 2 diabetes, and cardiovascular disease have become major social problems. We have previously shown that Astaxanthin (AX), which is a natural antioxidant, significantly ameliorates obesity-induced glucose intolerance and insulin resistance. It is well known that AX is a strong lipophilic antioxidant and has been shown to be beneficial for acute inflammation. However, the actual effects of AX on chronic inflammation in adipose tissue (AT) remain unclear. To observe the effects of AX on AT functions in obese mice, we fed six-week-old male C57BL/6J on high-fat-diet (HFD) supplemented with or without 0.02% of AX for 24 weeks. We determined the effect of AX at 10 and 24 weeks of HFD with or without AX on various parameters including insulin sensitivity, glucose tolerance, inflammation, and mitochondrial function in adipose tissue. We found that AX significantly reduced oxidative stress and macrophage infiltration into AT, as well as maintaining healthy AT function. Furthermore, AX prevented pathological AT remodeling probably caused by hypoxia in AT. Collectively, AX treatment exerted anti-inflammatory effects via its antioxidant activity in AT, maintained the vascular structure of AT and preserved the stem cells and progenitor's niche, and enhanced anti-inflammatory hypoxia induction factor-2α-dominant hypoxic response. Through these mechanisms of action, it prevented the pathological remodeling of AT and maintained its integrity. Citation: Nawaz, A.; Nishida, Y.; Takikawa, A.; Fujisaka, S.; Kado, T.; Aminuddin, A.; Bilal, M.; Jeelani, I.; Aslam, M.R.; Nishimura, A.; et al. Astaxanthin, a Marine Carotenoid, Maintains the Tolerance and Integrity of Adipose Tissue and Contributes to Its Healthy Functions. Nutrients 2021, 13, 4374.
Various types of fish roe products are manufactured and consumed in Japan. Common products are salted or spicy seasoned walleye pollock roe, salted salmon roe, salted herring roe, salted-dried mullet roe, and flying fish roe. Although the raw materials are mostly imported, most of the products are produced in Japan, except for salmon roe products. All the products are distributed through chilled or frozen cold-chain with an estimated total of more than 70,000 ton/year. This chapter introduces the overall information about fish roe products and provides an overview of manufacture and storage methods, factors affecting quality, nutritional value, and hygiene control to reduce food risks. Fish roe allergies showing high frequency in Japan is also explained.
Objective The aim of the study is to investigate the therapeutic effects of astaxanthin (AST) and resveratrol (RVT) on multiorgan damage in an animal model of the supraceliac aortic ischemia-reperfusion (I/R). Methods In this study, 28 rats (n = 7/group), 200 to 250 g in weight, were randomized to four groups (1: Sham, 2: Control + I/R, 3: AST + I/R, and 4: RVT + I/R). Following the abdominal incision, aortic dissection was performed in the sham group without injury. Other groups underwent I/R injury via supraceliac aortic clamping (20 minutes) and reperfusion. The rats were administered olive oil (3 mL/kg) orally for 2 weeks before and 1 week after the laparotomy. Additionally, oral AST (10 mg/kg) or RVT (50 mg/kg) was given to the study groups. All rats were sacrificed on the 3rd week of the experiment after blood samples were taken for analysis. Multiple rat tissues were removed. Results We found that RVT increased total antioxidant status (TAS) and superoxide dismutase (SOD) levels, and decreased total oxidant status (TOS), oxidative stress index (OSI), myeloperoxidase (MPO), and malondialdehyde (MDA) levels, while AST increased the levels of TAS, decreased TNF-α, MDA, TOS, and OSI (p <0.05). Pathological investigations of the rat tissues revealed that both AST and RVT ameliorated tissue damage and apoptosis. Conclusion Our study suggests that AST and RVT might show therapeutic effects against oxidative tissue damage and apoptosis in an animal model of aortic I/R. Further studies are required. Key Points
Haematococcus lacustris is a green unicellular alga known for its diverse beneficial carotenoid, such as β-carotene, canthaxanthin, and astaxanthin, with varying compositions at different morphological stages. Each carotenoid might affect the lifespan of Caenorhabditis elegans by inhibiting reactive oxygen species (ROS). C. elegans were fed with carotenoids either using the Escherichia coli OP50 that had pre-consumed commercial carotenoids and the optimal Haematococcus medium containing a dissolved carotenoid. As a result, lifespan of C. elegans was extended by 1.3 times. Three carotenoids indirectly protected C. elegans against ROS by upregulating the expression of superoxide dismutase (maximum 9.8 times) and catalase (maximum 2.5 times). Carotenoids reduced the concentration of malondialdehyde, a product of lipid peroxidation induced by ROS, by 11.2 times in C. elegans. These results suggest that carotenoids could induce antioxidation effect and prolong the lifespan of C. elegans. And H. lacustris could be the most versatile producer of carotenoids.
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Human blood plasma and freshly isolated LDL were exposed to singlet oxygen (1O2) by thermal decomposition of synthetic endoperoxides. Exposure of blood plasma to 20 mM water-soluble 1O2 generator resulted in the depletion of ascorbate (100%), urate (75%), ubiquinol-10 (65%), protein thiols (50%), and bilirubin (25%), whereas under these conditions the levels of alpha-tocopherol, beta-carotene, and lycopene remained unchanged. The following rates of depletion were obtained by kinetic analysis (moles depleted per 100 mol of 1O2 consumed): protein thiols (5), urate (5), ascorbate (4), bilirubin (1), and ubiquinol-10 (0.008). In contrast, the rates of depletion using the lipid-soluble 1O2 generator were faster for bilirubin (13-fold), protein thiols (9-fold), ubiquinol-10 (8-fold), and ascorbate (5-fold), and slower for urate (2-fold). The formation of lipid hydroperoxides, including mostly cholesteryl linoleate hydroperoxide, was observed in 1O2-treated plasma (0.007-0.009 mol/100 mol 1O2) and LDL solutions (0.086 mol/100 mol 1O2). Based on competition kinetics, we estimate that 98% of 1O2 generated in the aqueous phase of plasma is quenched by components in this phase, mostly by plasma protein (63%; 6% by protein thiols), urate (9%; 5% by chemical quenching), and bilirubin (5%; 1% by chemical quenching). Ascorbate and ubiquinol-10 do not contribute to 1O2 quenching in plasma, and their oxidation is probably mediated secondary species. The remaining 1O2 generated in plasma (2%) diffuses into lipoprotein leading to the formation of lipid hydroperoxides with an efficiency of about 100-fold greater than that compared to aqueous generated 1O2. The principal 1O2 quenchers in LDL include apoB (42%), lycopene and beta-carotene (40%), and alpha-tocopherol (17%). The importance of carotenoids in the quenching of 1O2 in lipoprotein suggest that the beneficial effects of these compounds in health may in part be due to the elimination of this species in biology and medicine.
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Glucose and lipid metabolic parameters play crucial roles in metabolic syndrome and its major feature of insulin resistance. This study was designed to investigate whether dietary astaxanthin oil (ASX-O) has potential effects on metabolic syndrome features in an SHR/NDmcr-cp (cp/cp) rat model. Oral administration of ASX (50 mg/kg/day) for 22 weeks induced a significant reduction in arterial blood pressure in SHRcp. It also significantly reduced the fasting blood glucose level, homeostasis index of insulin resistance (HOMA-IR), and improved insulin sensitivity. The results also showed an improved adiponectin level, a significant increase in high-density lipoprotein cholesterol, a significant decrease in plasma levels of triglycerides, and non-esterified fatty acids. Additionally, ASX showed significant effects on the white adipose tissue by decreasing the size of the fat cells. These results suggest that ASX ameliorates insulin resistance by mechanisms involving the increase of glucose uptake, and by modulating the level of circulating lipid metabolites and adiponectin.
To understand the roles of carotenoids as singlet oxygen quenchers in marine organisms, quenching activities of eight major carotenoids, astaxanthin, canthaxanthin, β-carotene, zeaxanthin, lutein, tunaxanthin, fucoxanthin and halocynthiaxanthin were examined according to the method using a thermodissociable endoperoxide of 1,4-dimethylnaphthalene as a singlet oxygen generator. The second-order rate constant for the singlet oxygen quenching activity by each carotenoid was determined, suggesting that an increasing number of conjugated double bonds in carotenoid was proportional to greater quenching activity. The quenching activity of each carotenoid was found to be approximately 40 to 600 times greater than that of α-tocopherol. The potency of these carotenoids suggests that they may play a role in protecting marine organisms from active oxygen species.
Marine animals produce astaxanthin which is a carotenoid and antioxidant. In this study we determined the in vitro and ex vivo effects of astaxanthin on LDL oxidation. The oxidation of LDL was measured in a 1 ml reaction system consisting of increasing concentrations of astaxanthin (12.5, 25.0, 50.0 microg/ml), 400 microM V-70 (2, 2'-azobis(4-methoxy-2, 4-dimethylvaleronitrile)), and LDL (70 microg/ml protein). Astaxanthin dose, dependently significantly prolonged the oxidation lag time (31.5, 45.4, 65.0 min) compared with the control (19.9 min). For the ex vivo study 24 volunteers (mean age 28.2 [SD 7.8] years) consumed astaxanthin at doses of 1.8, 3.6,14.4 and 21.6 mg per day for 14 days. No other changes were made in the diet. Fasting venous blood samples were taken at days 0, +14. LDL lag time was longer (5.0, 26.2, 42.3 and 30.7% respectively) compared with day 0 after consuming astaxanthin at doses of 1.8, 3.6,14.4 and 21.6 mg for 14 days compared with day 0, but there was no difference in oxidation of LDL between day 0 (lag time 59.9+/-7.2 min) and day 14 (57.2+/-6.0 min) in the control group. Our results provide evidence that consumption of marine animals producing astaxanthin inhibits LDL oxidation and possibly therefore contributes to the prevention of atherosclerosis.
We investigated the effects of a dietary astaxanthin (ASX-O) on oxidative parameters in spontaneously hypertensive rats (SHR), by determination of the level of nitric oxide (NO) end products nitrite/nitrate (NO2-/NO3-) and lipid peroxidation in ASX-O-treated SHR. Oral administration of the ASX-O significantly reduced the plasma level of NO2-/NO3- compared to the control vehicle (p<0.05). The lipid peroxidation level, however, was reduced in both ASX-O- and olive oil-treated groups. We also analyzed the post-treatment effects of ASX-O on the vascular tissues by examining the changes in the aorta and coronary arteries and arterioles. The dietary ASX-O showed significant reduction in the elastin bands in the rat aorta (p<0.05). It also significantly decreased the [wall : lumen] aerial ratio of the coronary arteries. These results suggest that ASX-O can modulate the oxidative condition and may improve vascular elastin and arterial wall thickness in hypertension.
City University for supplying β-carotene. We are also indebted to Dr. V. Wood and Ms. A. Miyashita of Fuji Chemical Industry Co
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