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Active-Italia: Astaxanthin: Sources, Extraction, Stability, Biological Activities and Its Commercial Applications

Astaxanthin: Sources, Extraction, Stability, Biological Activities and Its Commercial Applications

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There is currently much interest in biological active compounds derived from natural resources, especially compounds that can efficiently act on molecular targets, which are involved in various diseases. Astaxanthin (3,3′-dihydroxy-β, β′-carotene-4,4′-dione) is a xanthophyll carotenoid, contained in Haematococcus pluvialis, Chlorella zofingiensis, Chlorococcum, and Phaffia rhodozyma. It accumulates up to 3.8% on the dry weight basis in H. pluvialis.

Abstract
Our recent published data on astaxanthin extraction, analysis, stability studies, and its biological activities results were added to this review paper.

Based on our results and current literature, astaxanthin showed potential biological activity in in vitro and in vivo models.

These studies emphasize the influence of astaxanthin and its beneficial effects on the metabolism in animals and humans. Bioavailability of astaxanthin in animals was enhanced after feeding Haematococcus biomass as a source of astaxanthin.

Astaxanthin, used as a nutritional supplement, antioxidant and anticancer agent, prevents diabetes, cardiovascular diseases, and neurodegenerative disorders, and also stimulates immunization.

Astaxanthin products are used for commercial applications in the dosage forms as tablets, capsules, syrups, oils, soft gels, creams, biomass and granulated powders.

Astaxanthin patent applications are available in food, feed and nutraceutical applications. The current review provides up-to-date information on astaxanthin sources, extraction, analysis, stability, biological activities, health benefits and special attention paid to its commercial applications.

Introduction
Astaxanthin is a xanthophyll carotenoid which is found in various microorganisms and marine animals.

It is a red fat-soluble pigment which does not have pro-Vitamin A activity in the human body, although some of the studies reported that astaxanthin has more potent biological activity than other carotenoids.

The United States Food and Drug Administration (USFDA) has approved the use of astaxanthin as food colorant in animal and fish feed.

The European Commission considers natural astaxanthin as a food dye. Haematococcus pluvialis is a green microalga, which accumulates high astaxanthin content under stress conditions such as high salinity, nitrogen deficiency, high temperature and light.

Astaxanthin produced from H. pluvialis is a main source for human consumption. It is used as a source of pigment in the feed for salmon, trout and shrimp.

For dietary supplement in humans and animals, astaxanthin is obtained from seafood or extracted from H. pluvialis.

The consumption of astaxanthin can prevent or reduce risk of various disorders in humans and animals. The effects of astaxanthin on human health nutrition have been published by various authors.

In our previous reviews, we included recent findings on the potential effects of astaxanthin and its esters on biological activities.

The use of astaxanthin as a nutritional supplement has been rapidly growing in foods, feeds, nutraceuticals and pharmaceuticals.

This present review paper provides information on astaxanthin sources, extraction methods, storage stability, biological activities, and health benefits for the prevention of various diseases and use in commercial applications.

(…) 8. Biological Activities of Astaxanthin and Its Health Benefits
8.1. Antioxidant Effects
An antioxidant is a molecule which can inhibit oxidation. Oxidative damage is initiated by free radicals and reactive oxygen species (ROS).

These molecules have very high reactivity and are produced by normal aerobic metabolism in organisms. Excess oxidative molecules may react with proteins, lipids and DNA through chain reaction, to cause protein and lipid oxidation and DNA damage which are associated with various disorders. This type of oxidative molecules can be inhibited by endogenous and exogenous antioxidants such as carotenoids.

Carotenoids contain polyene chain, long conjugated double bonds, which carry out antioxidant activities by quenching singlet oxygen and scavenging radicals to terminate chain reactions.

The biological benefits of carotenoids may be due to their antioxidant properties attributed to their physical and chemical interactions with cell membranes. Astaxanthin had higher antioxidant activity when compared to various carotenoids such as lutein, lycopene, α-carotene and β-carotene reported by Naguib et al.

The antioxidant enzymes catalase, superoxide dismutase, peroxidase and thiobarbituric acid reactive substances (TBARS) were high in rat plasma and liver after feeding Haematococcus biomass as source of astaxanthin.

Astaxanthin in H. pluvialis offered the best protection from free radicals in rats followed by β-carotene and lutein. Astaxanthin contains a unique molecular structure in the presence of hydroxyl and keto moieties on each ionone ring, which are responsible for the high antioxidant properties.

Antioxidant activity of astaxanthin was 10 times more than zeaxanthin, lutein, canthaxanthin, β-carotene and 100 times higher than α-tocopherol.

The oxo functional group in carotenoids has higher antioxidant activity without pro-oxidative contribution.

The polyene chain in astaxanthin traps radicals in the cell membrane, while the terminal ring of astaxanthin could scavenge radicals at the outer and inner parts of cell membrane.

Antioxidant enzyme activities were evaluated in the serum after astaxanthin was supplemented in the diet of rabbits, showing enhanced activity of superoxide dismutase and thioredoxin reductase whereas paraoxonase was inhibited in the oxidative-induced rabbits.

Antioxidant enzyme levels were increased when astaxanthin fed to ethanol-induced gastric ulcer rats.

8.2. Anti-Lipid Peroxidation Activity
Astaxanthin has a unique molecular structure which enables it to stay both in and outside the cell membrane. It gives better protection than β-carotene and Vitamin C which can be positioned inside the lipid bilayer. It serves as a safeguard against oxidative damage by various mechanisms, like quenching of singlet oxygen; scavenging of radicals to prevent chain reactions; preservation of membrane structure by inhibiting lipid peroxidation; enhancement of immune system function and regulation of gene expression.

Astaxanthin and its esters showed 80% anti-lipid peroxidation activity in ethanol induced gastric ulcer rats and skin cancer rats. Astaxanthin inhibited lipid peroxidation in biological samples reported by various authors.

8.3. Anti-Inflammation
Astaxanthin is a potent antioxidant to terminate the induction of inflammation in biological systems.

Astaxanthin acts against inflammation. Algal cell extracts of Haematococcus and Chlorococcum significantly reduced bacterial load and gastric inflammation in H. pylori-infected mice. Park et al. reported astaxanthin reduced the DNA oxidative damage biomarker inflammation, thus enhancing immune response in young healthy adult female human subjects.

Haines et al. reported lowered bronchoalveolar lavage fluid inflammatory cell numbers, and enhanced cAMP, cGMP levels in lung tissues after feeding astaxanthin with Ginkgo biloba extract and Vitamin C.

Another study showed astaxanthin esters and total carotenoids from Haematococcus exerted a dose-dependent gastroprotective effect on acute, gastric lesions in ethanol-induced gastric ulcers in rats.

This may be due to inhibition of H1, K1 ATPase, upregulation of mucin content and an increase in antioxidant activities. Astaxanthin showed protective effect on high glucose induced oxidative stress, inflammation and apoptosis in proximal tubular epithelial cells.

Astaxanthin is a promising molecule for the treatment of ocular inflammation in eyes as reported by the Japanese researchers.

Astaxanthin can prevent skin thickening and reduce collagen reduction against UV induced skin damage.

8.4. Anti-Diabetic Activity
Generally, oxidative stress levels are very high in diabetes mellitus patients. It is induced by hyperglycemia, due to the dysfunction of pancreatic β-cells and tissue damage in patients.

Astaxanthin could reduce the oxidative stress caused by hyperglycemia in pancreatic β-cells and also improve glucose and serum insulin levels.

Astaxanthin can protect pancreatic β-cells against glucose toxicity. It was also shown to be a good immunological agent in the recovery of lymphocyte dysfunctions associated with diabetic rats.

In another study, ameliorate oxidative stress in streptozotocin-diabetes rats were inhibited by the combination of astaxanthin with α-tocopherol.

It is also inhibited glycation and glycated protein induced cytotoxicity in human umbilical vein endothelial cells by preventing lipid/protein oxidation.

Improved insulin sensitivity in both spontaneously hypertensive corpulent rats and mice on high fat plus high fructose diets was observed after feeding with astaxanthin.

The urinary albumin level in astaxanthin treated diabetic mice was significantly lower than the control group.

Some of the studies demonstrated that astaxanthin prevents diabetic nephropathy by reduction of the oxidative stress and renal cell damage.

8.5. Cardiovascular Disease Prevention
Astaxanthin is a potent antioxidant with anti-inflammatory activity and its effect examined in both experimental animals and human subjects.

Oxidative stress and inflammation are pathophysiological features of atherosclerotic cardiovascular disease.

Astaxanthin is a potential therapeutic agent against atherosclerotic cardiovascular disease.

The efficacy of disodium disuccinate astaxanthin (DDA) in protecting mycocardium using mycocardial ischemia reperfusion model in animals was evaluated.

Myocardial infarct size was reduced in Sprague Dawley rats, and improved in myocardial salvage in rabbits after four days of pre-treatment with DDA at 25, 50 and 75 mg/kg body weight [89,90].

Astaxanthin was found in rat mycocardial tissues after pretreatment with DDA at dosage of 150 and 500 mg/kg/day for seven days.

Astaxanthin effects on blood pressure in spontaneously hypertensive rats (SHR), normotensive Wistar Kyoto rats (NWKR) and stroke prone spontaneously hypertensive rats (SPSHR) were reported.

Astaxanthin was found in the plasma, heart, liver, platelets, and increased basal arterial blood flow in mice fed with astaxanthin derivative.

Human umbilical vien endothelial cells and platelets treated with the astaxanthin showed increased nitric oxide levels and decrease in peroxynitrite levels.

Mice fed 0.08% astaxanthin had higher heart mitochondrial membrane potential and contractility index compared to the control group.

Astaxanthin effects on paraoxonase, thioredoxin reductase activities, oxidative stress parameters and lipid profile in hypercholesterolemic rabbits were evaluated. Astaxanthin prevented the activities of those enzymes from hypercholesterolemia induced protein oxidation at the dosages of 100 mg and 500 mg/100 g.

8.6. Anticancer Activity
The specific antioxidant dose may be helpful for the early detection of various degenerative disorders.

Reactive oxygen species such as superoxide, hydrogen peroxide and hydroxyl radical are generated in normal aerobic metabolism. Singlet oxygen is generated by photochemical events whereas peroxyl radicals are produced by lipid peroxidation.

These oxidants contribute to aging and degenerative diseases such as cancer and atherosclerosis through oxidation of DNA, proteins and lipids.

Antioxidant compounds decrease mutagenesis and carcinogenesis by inhibiting oxidative damage to cells.

Cell–cell communication through gap junctions is lacking in human tumors and its restoration tends to decrease tumor cell proliferation.

Gap junctional communication occurs due to an increase in the connexin-43 protein via upregulation of the connexin-43 gene.

Gap junctional communication was improved in between the cells by natural carotenoids and retinoids.

Canthaxanthin and astaxanthin derivatives enhanced gap junctional communication between mouse embryo fibroblasts.

Increased connexin-43 expression in murine fibroblast cells by β-carotene was reported.

Astaxanthin showed significant antitumor activity when compared to other carotenoids like canthaxanthin and β-carotene.

It also inhibited the growth of fibrosarcoma, breast, and prostate cancer cells and embryonic fibroblasts.

Increased gap junctional intercellular communication in primary human skin fibroblasts cells were observed when treated with astaxanthin.

Astaxanthin inhibited cell death, cell proliferation and mammary tumors in chemically induced male/female rats and mice.

H. pluvialis extract inhibited the growth of human colon cancer cells by arresting cell cycle progression and promoting apoptosis reported by Palozza et al.

Nitroastaxanthin and 15-nitroastaxanthin are the products of astaxanthin with peroxynitrite, 15-nitroastaxanthin anticancer properties were evaluated in a mouse model. Epstein-Barr virus and carcinogenesis in mouse skin papillomas were significantly inhibited by astaxanthin treatment.

8.7. Immuno-Modulation
Immune system cells are very sensitive to free radical damage.

The cell membrane contains poly unsaturated fatty acids (PUFA). Antioxidants in particular astaxanthin offer protection against free radical damage to preserve immune-system defenses.

There are reports on astaxanthin and its effect on immunity in animals under laboratory conditions however clinical research is lacking in humans. Astaxanthin showed higher immuno-modulating effects in mouse model when compared to β-carotene.

Enhanced antibody production and decreased humoral immune response in older animals after dietary supplementation of astaxanthin was reported.

Astaxanthin produced immunoglobulins in human cells in a laboratory study.

Eight week-supplementation of astaxanthin in humans resulted in increased blood levels of astaxanthin and improved activity of natural killer cells which targeted and destroyed cells infected with viruses.

In this study, T and B cells were increased, DNA damage was low, and C-reactive protein (CRP) was significantly lower in the astaxanthin supplemented group.

Source of the Article

Ranga Rao Ambati, Siew-Moi Phang, Sarada Ravi and Ravishankar Gokare Aswathanarayana.

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