ArticlePDF Available

Antioxidant effect of astaxanthin on phospholipid peroxidation in human erythrocytes


Abstract and Figures

Phospholipid hydroperoxides (PLOOH) accumulate abnormally in the erythrocytes of dementia patients, and dietary xanthophylls (polar carotenoids such as astaxanthin) are hypothesised to prevent the accumulation. In the present study, we conducted a randomised, double-blind, placebo-controlled human trial to assess the efficacy of 12-week astaxanthin supplementation (6 or 12 mg/d) on both astaxanthin and PLOOH levels in the erythrocytes of thirty middle-aged and senior subjects. After 12 weeks of treatment, erythrocyte astaxanthin concentrations were higher in both the 6 and 12 mg astaxanthin groups than in the placebo group. In contrast, erythrocyte PLOOH concentrations were lower in the astaxanthin groups than in the placebo group. In the plasma, somewhat lower PLOOH levels were found after astaxanthin treatment. These results suggest that astaxanthin supplementation results in improved erythrocyte antioxidant status and decreased PLOOH levels, which may contribute to the prevention of dementia.
Content may be subject to copyright.
Antioxidant effect of astaxanthin on phospholipid peroxidation
in human erythrocytes
Kiyotaka Nakagawa
*, Takehiro Kiko
, Taiki Miyazawa
, Gregor Carpentero Burdeos
Fumiko Kimura
, Akira Satoh
and Teruo Miyazawa
Food and Biodynamic Chemistry Laboratory, Graduate School of Agricultural Science, Tohoku University,
Sendai 981-8555, Japan
Life Science Institute, Yamaha Motor Company Limited, Shizuoka 437-0061, Japan
(Received 19 July 2010 Revised 25 November 2010 Accepted 26 November 2010 First published online 31 January 2011)
Phospholipid hydroperoxides (PLOOH) accumulate abnormally in the erythrocytes of dementia patients, and dietary xanthophylls
(polar carotenoids such as astaxanthin) are hypothesised to prevent the accumulation. In the present study, we conducted a randomised,
double-blind, placebo-controlled human trial to assess the efficacy of 12-week astaxanthin supplementation (6 or 12 mg/d) on both astax-
anthin and PLOOH levels in the erythrocytes of thirty middle-aged and senior subjects. After 12 weeks of treatment, erythrocyte astaxanthin
concentrations were higher in both the 6 and 12 mg astaxanthin groups than in the placebo group. In contrast, erythrocyte PLOOH
concentrations were lower in the astaxanthin groups than in the placebo group. In the plasma, somewhat lower PLOOH levels were
found after astaxanthin treatment. These results suggest that astaxanthin supplementation results in improved erythrocyte antioxidant
status and decreased PLOOH levels, which may contribute to the prevention of dementia.
Key words: Astaxanthin: Phospholipid hydroperoxides: Erythrocytes: Dementia
We have previously confirmed that higher levels of
phospholipid hydroperoxides (PLOOH), the primary
oxidation products of phospholipids (PL)
, accumulate
abnormally in the erythrocytes of dementia patients
. Such
erythrocytes with high levels of lipid hydroperoxides have
been postulated to have a decreased ability to transport
oxygen to the brain, which may impair blood rheology,
thus facilitating dementia
(4 8)
. Recently, we have developed
an HPLC method to determine erythrocyte carotenoid
. Using this method, we gathered evidence that
accumulation of polar oxygenated carotenoids (xanthophylls)
occurs predominantly in human erythrocytes
, and that a
decrease in xanthophylls and an increase in PLOOH levels in
erythrocytes correlate with the severity of dementia
. These
findings led to the hypothesis that xanthophyll supplementation
may minimise the accumulation of erythrocyte PLOOH, and that
xanthophylls could be used therapeutically as drugs or func-
tional foods to prevent the disease. Although there is still
scarce information on whether orally administered xantho-
phylls are distributed to human erythrocytes and actually inhibit
erythrocyte PLOOH formation, our recent human study has
revealed antioxidant properties of the xanthophyll lutein
towards erythrocyte PLOOH formation
. Animal studies
have also supported this hypothesis
Among xanthophylls, astaxanthin has recently received
attention for its potent antioxidant activity
. Astaxanthin
is naturally synthesised by plants and algae, and is now com-
mercially available as a food supplement from Haematococcus
. The recommended daily intake is estimated to be
112 mg/d; however, there is not much information regarding
the bioavailability of astaxanthin in humans. To the best of
our knowledge, the occurrence and antioxidant roles of astax-
anthin in human erythrocytes have not been reported.
In this investigation of whether administered astaxanthin is
distributed to erythrocytes and inhibits erythrocyte PLOOH
formation, we conducted a randomised, double-blind, pla-
cebo-controlled human trial. The efficacy of 12-week astax-
anthin supplementation (6 or 12 mg) on both astaxanthin
and PLOOH levels in the erythrocytes of thirty middle-aged
and senior subjects was investigated. For erythrocyte astax-
anthin analysis, a newly developed HPLC coupled with
tandem MS (MS/MS) method was applied. Our findings (the
inhibitory effect of astaxanthin on erythrocyte PLOOH)
* Corresponding author: K. Nakagawa, fax þ 81 22 717 8905, email
Abbreviations: CL, chemiluminescence; MRM, multiple reaction monitoring; PCOOH, phosphatidylcholine hydroperoxide; PEOOH,
phosphatidylethanolamine hydroperoxide; PL, phospholipid; PLOOH, phospholipid hydroperoxide.
British Journal of Nutrition (2011), 105, 1563–1571 doi:10.1017/S0007114510005398
q The Authors 2011
British Journal of Nutrition
would provide new insights into the possible application of
astaxanthin as an anti-dementia agent.
Subjects and methods
Subjects and materials
The present study was conducted according to the
guidelines laid down in the Declaration of Helsinki, and all
procedures involving human subjects were approved by the
ethics committee of Anti-Aging Science (Tokyo, Japan; ethics
no. I030807). All subjects were recruited from the Anti-Aging
Science volunteer database, and written informed consent
was obtained from all subjects. Exclusion criteria included
pregnancy, lactation and severe medical illness.
Two doses of the test materials were prepared by filling soft
capsules with astaxanthin-rich Haematococcus pluvialis oil
; Yamaha Motor Company, Limited, Shizuoka,
. Compositions of the test materials were 75 mg Pure-
sta oil 80 and 145 mg olive oil/capsule for the low dose (equiv-
alent to 6 mg astaxanthin dialcohol); and 150 mg Puresta oil 80
and 70 mg olive oil/capsule for the high dose (equivalent to
12 mg astaxanthin dialcohol). To prepare placebo capsules,
150 mg maize oil was used instead of 150 mg Puresta oil 80.
Placebo capsules were coloured to appear identical to test
Supplementation trial
A 12-week randomised, double-blind, placebo-controlled trial
was conducted. A total of thirty healthy subjects (fifteen men
and fifteen women), between 50 and 69 years of age (mean
56·3 (
SD 5·3) years), with a BMI of 27·5 (SD 2·1) kg/m
recruited, and randomly received 0 (placebo), 6 or 12 mg
astaxanthin. During the 12-week trial, subjects ingested one
of the three astaxanthin-dosed (0, 6 or 12 mg) capsules with
an appropriate amount of water once daily after breakfast.
Before and after the supplementation period (weeks 0 and
12, respectively), anthropometric data (e.g. height, body
weight and blood pressure) and blood samples were collected
from the subjects after they had fasted overnight, adverse
effects were assessed by interviews and self-reports, and com-
pliance was checked by self-reports and returned capsule
counts. Throughout the study period, subjects were instructed
to maintain their usual lifestyle (avoid excessive eating and
drinking, intense exercise and lack of sleep). Dietary intake,
alcohol consumption and physical activity (pedometer
count) during the 3 d before each blood collection (weeks 0
and 12) were also assessed by self-reports.
Measurement of erythrocyte astaxanthin and other
Blood samples were subjected to centrifugation at 1000 g
for 10 min at 48C. After the plasma and buffy coat were removed,
erythrocytes were washed three times with PBS (pH 7·4) to pre-
pare packed cells. For the determination of erythrocyte caroten-
oids (including astaxanthin), packed cells (2·5 ml) were diluted
with 2·5 ml of water and were mixed with 5 ml of 80 m
ethanolic pyrogallol, 1·0 ml of 1·8
M-aqueous KOH and 40 ml
of 1 m
M-ethanolic echinenone (internal standard)
. After the
addition of 1·25 ml of 0·1
M-aqueous SDS, the sample was
mixed with 15 ml of hexanedichloromethane (5:1) to extract
erythrocyte carotenoids. The extract was purified by a Sep-
Pak silica cartridge (Waters, Milford, MA, USA), and then was
subjected to HPLC-MS/MS for the determination of astaxanthin.
The HPLC-MS/MS apparatus consisted of a liquid chromato-
graph (Shimadzu, Kyoto, Japan) and a 4000 QTRAP MS/MS
instrument (Applied Biosystems, Foster City, CA, USA). The
MS/MS parameters (e.g. collision energy) were optimised with
an astaxanthin standard under positive atmospheric pressure
chemical ionisation. The standard and erythrocyte extracts
were separated with a C30 carotenoid column (4·6 £ 250 mm,
5 mm; YMC, Kyoto, Japan). The column was eluted using a
binary gradient consisting of the following HPLC solvents: A,
methanolmethyl tert-butyl ether water (83:15:2; containing
M-ammonium acetate); B, methanolmethyl tert-butyl
etherwater (8:90:2; containing 2·6
M-ammonium acetate).
The gradient profile was as follows: 0 12 min, 1045 % B
linear; 1224 min, 45100 % B linear; 24 38 min, 100 % B. The
flow rate was adjusted to 1 ml/min, and the column temperature
was maintained at 258C. Astaxanthin was detected in the post-
column by MS/MS with multiple reaction monitoring (MRM)
for the transition of the parent ion to the product ion. The
concentration of erythrocyte astaxanthin was calculated
using the standard curve of astaxanthin and was adjusted by
the percentage recovery of the added echinenone (internal
standard). For the determination of other carotenoids,
erythrocyte extracts were analysed by HPLC coupled with
UV diode array detection and atmospheric pressure chemical
ionisation MS
Measurement of erythrocyte phospholipid hydroperoxides
For the determination of erythrocyte PLOOH
(1 3)
, total lipids
were extracted from packed cells with a mixture of 2-propanol
and chloroform containing butylated hydroxytoluene. PLOOH
(i.e. phosphatidylcholine hydroperoxide (PCOOH) and phos-
phatidylethanolamine hydroperoxide (PEOOH)) in the total
lipids were measured by HPLC with chemiluminescence
(CL) detection. The column was a 4·6 £ 250 mm, 5 mm Fine-
pak SIL NH2-5 (Japan Spectroscopic Company, Tokyo,
Japan), the eluent was 2-propanolmethanolwater
(135:45:20), and the flow rate was 1 ml/min. Post-column CL
detection was carried out using a CLD-100 detector (Tohoku
Electronic Industries Company, Sendai, Japan). A mixture of
luminol and cytochrome c in 50 m
M-borate buffer (pH 10·0)
was used as a hydroperoxide-specific post-column CL reagent.
Calibration was carried out using PCOOH or PEOOH
Other biochemical measurements
For plasma samples, astaxanthin, other carotenoids and
PLOOH were determined by HPLC-MS/MS, HPLC-UV
(1 3)
, respectively. Tocopherols in erythrocytes
K. Nakagawa et al.1564
British Journal of Nutrition
and plasma were measured by HPLC with fluorescence
. Also, blood chemistries such as haematological
and blood biochemical parameters were analysed using
standardised methods.
Statistical analysis
Data are expressed as means and standard deviations.
One-way ANOVA was used to compare means among
the three groups. If a statistically significant difference
(P, 0·05) was detected, Dunnett’s test was performed for com-
parison between the control group and one of the two astax-
anthin groups. For comparison of baseline (week 0) and
post-dosing (week 12) values in each treatment group, a
paired t test was used. These statistical analyses were done
using SPSS for Windows (SPSS Inc., Chicago, IL, USA).
As mentioned in the introduction, the distribution of astax-
anthin in erythrocytes has not been reported, mainly due
to the lack of an analytical method. We, therefore, developed
an HPLC-MS/MS method to analyse for erythrocyte astax-
anthin before we conducted the 12-week astaxanthin sup-
plementation study. In brief, an astaxanthin standard was
analysed by MS/MS with flow injection, and astaxanthin
showed an intense molecular ion at m/z 597 (M þ H)
Product ion scanning was conducted for the ion and astax-
anthin-specific fragment ions (e.g. m/z 147) were identified.
These ions (m/z 597 and 147) allowed us to quantitatively
determine erythrocyte astaxanthin concentrations using
HPLC-MS/MS with MRM (Fig. 1(a)).
In the human trial, before administration (baseline),
there were no significant differences in anthropometric para-
meters among the three groups (Table 1). The trial was
completed without any of the subjects withdrawing. The
mean compliance to the prescribed dose for the 6 mg astax-
anthin, 12 mg astaxanthin and placebo groups was 99·5
SD 3·7), 98·1 (SD 1·9) and 98·7 (SD 2·4) %, respectively. The
average energy intake during the 12-week trial, as calculated
from the self-reports, did not statistically differ among the
groups. Furthermore, no significant differences were observed
among the groups in the intake of each type of nutrient
(carbohydrate, protein, fat, cholesterol and fibre), alcohol
consumption and pedometer counts. Data from all thirty
subjects were, therefore, included in the statistical analysis.
The results of the physical, haematological and blood bio-
chemical measurements before and after 12 weeks of dosing
are shown in Tables 2 and 3. Some parameters (i.e. Hb, hae-
matocrit, mean corpuscular volume, mean corpuscular Hb,
uric acid, total cholesterol, LDL, fasting glucose and Hb
showed changes from baseline in the astaxanthin groups
Time (min)
Relative intensity (%)
10 20 30
Time (min)
Relative intensity (%)
Fig. 1. Typical multiple reaction monitoring (MRM) and chemiluminescence (CL) chromatograms of (a) astaxanthin and (b) phospholipid hydroperoxides (PLOOH)
in erythrocytes taken (
) before and ( ) after 12-week supplementation of astaxanthin (12 mg/d). Erythrocyte astaxanthin and PLOOH were determined
by HPLC-MS/MS with MRM and HPLC-CL, respectively. Peak identifications are as follows: 1, astaxanthin; 2, phosphatidylcholine hydroperoxide; 3, phosphatidy-
lethanolamine hydroperoxide.
Table 1. Baseline characteristics of the study subjects in the 0 (placebo), 6 and 12 mg astaxanthin groups
(Mean values and standard deviations)
0 mg 6 mg 12 mg
Background factors Mean
SD Mean SD Mean SD P *
Age (years) 56·6 4·4 56·3 6·6 56·1 5·1 0·979
Total number of subjects 10 10 10
Men 5 5 5
Women 5 5 5
Height (cm) 159 11 160 8 164 7 0·407
Weight (kg) 70·3 9·3 70·5 8·1 74·4 5·3 0·429
BMI (kg/m
) 27·7 2·1 27·4 2·2 27·6 2·1 0·946
Systolic blood pressure (mmHg) 133 92 124 16 134 18 0·371
Diastolic blood pressure (mmHg) 83·1 10·9 82·3 9·8 90·7 15·0 0·250
* One-way ANOVA test among groups.
Astaxanthin inhibits lipid peroxidation 1565
British Journal of Nutrition
and/or the placebo group. However, these changes were
small and were observed to be within the normal range irre-
spective of the test materials administered. Moreover, no
differences were noted among the three groups in any par-
ameters at baseline or post-dosing. Therefore, the physical
and metabolic states of the subjects were considered to be
randomised homogeneously throughout the trial.
In the typical MRM chromatogram of the erythrocyte extract
taken before and 12 weeks after supplementation, astaxanthin
was clearly detected (Fig. 1(a)). After supplementation, the
erythrocyte astaxanthin concentration significantly increased
and was higher than that of the placebo group (Table 4).
On the other hand, in a typical CL chromatogram of erythro-
cyte total lipids, PCOOH and PEOOH were identified as the
predominant PLOOH forms (Fig. 1(b)). After supplementation,
erythrocyte PLOOH concentration decreased and was lower
than that of the placebo group (Table 4). In plasma, the
only detectable PLOOH was PCOOH, and a somewhat
lower PCOOH level was found after astaxanthin supplemen-
tation (Table 5). In both erythrocytes and plasma, astaxanthin
supplementation did not affect the levels of other carotenoids
(except for small changes in lycopene and lutein) and
tocopherols. These results suggest that upon ingestion of
astaxanthin, it is absorbed, distributed and accumulated in
erythrocytes, where it acts as an antioxidant molecule, thereby
reducing PLOOH, an index of oxidative stress.
In recent years, medical and nutritional experts have seriously
considered the antioxidant properties of food constituents,
since the reactive oxygen species-mediated peroxidation of
Table 2. Changes in physical and haematological parameters before and after the
12-week administration of 0, 6 or 12 mg astaxanthin
(Mean values and standard deviations)
0 mg 6 mg 12 mg
Parameters Mean
SD Mean SD Mean SD P
Weight (kg)
Before 70·3 9·3 70·5 8·1 74·4 5·3 0·429
After 69·0 9·2 69·6 7·5 74·3 5·9 0·248
BMI (kg/m
Before 27·7 2·1 27·4 2·2 27·6 2·1 0·946
After 27·1 2·2 27·1 2·2 27·6 2·5 0·872
Systolic blood pressure (mmHg)
Before 133 18 124 16 134 18 0·371
After 134 16 127 17 134 16 0·555
Diastolic blood pressure (mmHg)
Before 83·1 10·9 82·3 9·8 90·7 15·0 0·250
After 81·4 9·7 84·2 9·2 86·1 9·1 0·532
Heart rate (bpm)
Before 70·5 12·5 71·8 11·2 70·9 4·0 956
After 68·7 11·6 71·1 7·1 75·6 6·2 0·213
Leucocytes (£ 10
Before 4·6 0·6 5·4 1·0 5·7 1·7 0·096
After 4·9 0·8 5·1 0·8 6·1 1·9 0·112
Erythrocytes (£ 10
Before 4·5 0·4 4·7 0·3 4·7 0·4 0·587
After 4·5 0·4 4·7 0·3 4·7 0·5 0·375
Hb (g/l)
Before 144 11 145 11 142 27 0·916
After 137** 11 142 13 141 23 0·799
Haematocrit (%)
Before 44·9 3·1 45·4 3·8 44·1 6·9 0·839
After 42·8** 2·9 44·1 4·1 43·9 5·2 0·750
Corpuscular volume (fl)
Before 99·1 6·3 96·0 3·3 94·8 12·9 0·522
After 96·3** 4·7 94·7** 3·3 94·0 12·1 0·783
Corpuscular Hb (pg/cell)
Before 31·7 2·1 30·8 1·1 30·5 5·3 0·735
After 30·9** 1·7 30·4** 1·2 30·3* 5·1 0·193
Corpuscular Hb concentration (%)
Before 32·0 0·8 32·1 0·8 32·0 2·1 0·983
After 32·1 0·6 32·1 0·6 32·0 2·1 0·988
Platelets (£ l0
Before 237 47 225 41 261 67 0·320
After 212 55 228 28 264 82 0·151
bpm, Beats/min.
Mean values were significantly different in the paired t test between before and after astaxanthin
administration: *P , 0·05, **P, 0·01.
One-way ANOVA test among groups.
K. Nakagawa et al.1566
British Journal of Nutrition
biological molecules (e.g. lipids) has been postulated to
induce a variety of pathological events such as atherogenesis,
ageing and dementia. Although many in vitro studies on the
antioxidant properties of food constituents have been
reported, little is known about the biological functions of diet-
ary antioxidants in vivo (especially in humans), except for a
few major antioxidants (e.g. tocopherols and ascorbic acid).
Since the bioavailability of food constituents is limited by
their digestibility and metabolic fate, oral administration
trials are favoured in evaluating their biological functions.
The present randomised, double-blind, placebo-controlled
human trial shows that when human subjects ingest asta-
xanthin, it is absorbed, distributed and accumulated in
erythrocytes, where it exhibits antioxidative effects (inhibition
of erythrocyte PLOOH). It is interesting to note that the
antioxidative effect observed in the present study was
Table 3. Changes in blood biochemical parameters before and after the 12-week administration
of 0, 6 or 12 mg astaxanthin
(Mean values and standard deviations)
0 mg 6 mg 12 mg
Parameters Mean
SD Mean SD Mean SD P
Total protein (g/l)
Before 73·0 2 ·3 74·6 2·1 71·9 3·5 0·950
After 72·6 2·1 73·9 1·8 71·5 3·8 0·162
Albumin (g/l)
Before 45·0 1 ·6 45·7 1·1 44·2 2·2 0·161
After 44·6 1·0 45·4 1·1 43·8 2·2 0·081
Total bilirubin (mg/l)
Before 8·0 2·9 7·0 2·1 7·0 2·7 0·615
After 7·4 1·9 7·2 2·9 7·3 2·9 0·986
GOT (U/l)
Before 22·6 6 ·9 25·0 9·2 23·0 8·4 0·786
After 21·0 5·6 21·3 3·9 23·6 11·0 0·700
GPT (U/l)
Before 25·7 17·6 30·6 15·2 24·1 12·9 0·620
After 21·2 13·0 24 ·2 8·0 25·0 15·6 0·779
ALP (U/l)
Before 225 67 238 55 203 72 0·489
After 225 58 236 58 197 60 0·336
g-GTP (U/l)
Before 40·8 23·6 35·1 15·2 41·3 31·5 0·820
After 37·3 19·4 30·7 11·4 41·6 38·5 0·638
Urea (mg/l)
Before 129 25 144 31 146 24 0·340
After 139 29 128 25 135 29 0·671
Creatinine (mg/l)
Before 7·0 1·5 7·3 1·4 7·8 2·1 0·543
After 7·0 1·5 7·2 1·5 8·0 1·9 0·404
Uric acid (mg/l)
Before 52·0 11·8 62·6 19·2 57·8 16·8 0·357
After 53·1 10·4 57 ·9* 16·0 56·3 13·7 0·721
Total cholesterol (mg/l)
Before 2180 440 2260 390 2030 230 0·372
After 2040* 430 2150 320 1960 150 0·401
HDL (mg/l)
Before 588 147 688 190 588 121 0·268
After 581 130 655 147 562 105 0·250
LDL (mg/l)
Before 1350 350 1320 370 1170 200 0·396
After 1220* 370 1240 290 1120 190 0·629
TAG (mg/l)
Before 1250 520 1110 780 1250 730 0·869
After 1140 410 1160 670 1360 1140 0 ·790
Fasting glucose (mg/l)
Before 1010 60 1020 110 1010 60 0·948
After 1040** 50 1040 110 1050 110 0·985
Before 5·1 0·3 5·1 0·3 5·1 0·3 0·982
After 5·2 0·3 5·2 0·2 5·2** 0·3 1·000
GOT, glutamic oxaloacetic transaminase; GPT, glutamic pyruvic transaminase; ALP, alkaline phosphatase;
g-GTP, g-glutamic transpeptidase.
Mean values were significantly different in the paired t test between before and after astaxanthin administration:
*P, 0·05, **P, 0·01.
One-way ANOVA test among groups.
Astaxanthin inhibits lipid peroxidation 1567
British Journal of Nutrition
produced by a relatively short-term supplementation with
astaxanthin (12 weeks).
In the present study, since the distribution of astaxanthin
in erythrocytes had not previously been reported, we
developed an HPLC-MS/MS method to analyse the erythrocyte
astaxanthin content before we conducted the astaxanthin sup-
plementation study. Using MS/MS, we found that protonated
astaxanthin tended to generate product ions (e.g. m/z 147).
Table 4. Changes in carotenoids, phospholipid hydroperoxides (PLOOH) and tocopherol contents in
erythrocytes taken before and after the 12-week administration of 0, 6 or 12 mg astaxanthin
(Mean values and standard deviations)
0 mg 6 mg 12 mg
Parameters Mean
SD Mean SD Mean SD P
Carteonoids (pmol/ml packed cells)
Before 4·0 1·7 2·9 1·3 2·8 1·0 0·112
After 3·6 1·2 35·7**†† 11·1 44·9**†† 24·0 0·000
Before 43·3 6·8 39·8 7·5 39·5 6·1 0·391
After 43·2 8·5 42·1* 8·0 40·6 7·4 0·765
Before 7·8 2·1 7·1 1·4 7·6 2·1 0·662
After 7·8 2·6 6·9 1·2 7·4 1·8 0·603
Before 10·5 2·6 9·9 3·0 9 ·6 2·1 0·735
After 10·1 2·4 9·5 2·7 9·4 2·6 0·802
Before 1·2 0·4 0·9 0·4 1·1 0·3 0·234
After 1·1 0·4 0·9 0·3 1·1 0·3 0·151
Before 3·9 0·3 3·8 0·4 3·7 0·4 0·678
After 3·9 0·4 3·8 0·5 3·8 0·6 0·902
Before 0·7 0·1 0·6 0·1 0·7 0·2 0·231
After 0·7 0·1 0·6 0·1 0·6* 0·2 0·273
Before 65·5 10·3 59·5 11·1 59·5 9·1 0·407
After 64·7 11·4 94·2**†† 20·9 102·4**†† 20·6 0·000
Non-polar carotenoidsk
Before 5·7 0·6 5·3 0·7 5·5 0·7 0·350
After 5·7 0·7 5·3 0·6 5·5 0·9 0·421
Total carotenoids
Before 71·3 10·6 64·8 11·6 65·1 9·8 0·397
After 70·4 11·7 99·4**†† 21·5 107·9**†† 20·5 0·000
PLOOH (pmol/ml packed cells)
Before 8·8 6·6 8·7 4·6 12·3 7·0 0·340
After 9·1 6·3 5·2** 2·7 6·6** 2·3 0·122
Before 5·1 2·4 4·6 3·6 6·3 6·5 0·706
After 5·8 2·6 2·8*† 1·8 3·0† 2·5 0·011
Total PLOOH{
Before 13·9 7·6 13·3 6·1 18·6 11·6 0·353
After 14·9 8·3 8·0**† 3·8 9·7**† 4·0 0·031
Tocopherols (nmol/ml packed cells)
Before 7·8 2·4 7·2 2·6 6·9 1·0 0·663
After 8·4 1·9 7·8 0·9 7·6 0·8 0·365
Before 0·9 0·4 0·8 0·5 0·9 0·3 0·872
After 0·8 0·3 0·8 0·4 0·8 0·3 0·998
Total tocopherols
Before 8·7 2·3 8·0 2·6 7·8 1·1 0·652
After 9·2 2·0 8·6 1·0 8·4 0·9 0·372
PCOOH, phosphatidylcholine hydroperoxide; PEOOH, phosphatidylethanolamine hydroperoxide.
Mean values were significantly different in the paired t test between before and after astaxanthin administration:
*P, 0·05, **P, 0·01.
Mean values were significantly different in Dunnett’s test between the control group and one of the two astaxanthin
groups: P , 0·05, ††P, 0·01.
One-way ANOVA test among groups.
§ Xanthophylls are the sum of astaxanthin, lutein, zeaxanthin and b-cryptoxanthin.
k Non-polar carotenoids are sum of a-carotene, b-carotene and lycopene.
{ PLOOH are the sum of PCOOH and PEOOH.
K. Nakagawa et al.1568
British Journal of Nutrition
The product ion indicated that MRM could be adapted to the
HPLC-MS/MS analysis of astaxanthin. Under optimised con-
ditions, the detection limit of the HPLC-MS/MS with the
MRM method was very sensitive at 0·02 pmol astaxanthin/
injection. The characteristics and advantages of our HPLC-
MS/MS method are as follows. The method was selective
and sensitive enough to measure astaxanthin in erythrocytes
(Fig. 1(a)) as well as in the plasma. Also, the method was
sufficiently simple and convenient to be applicable to a
large number of samples. The method, therefore, would be
a powerful tool for studying the metabolic fate of astaxanthin
as well as its bioavailability.
Until now, there are few reports concerning human
erythrocyte carotenoids. Some studies have successfully
detected erythrocyte carotenoids (mainly b-carotene)
while other studies have been unable to detect these
Table 5. Changes in carotenoids, phospholipid hydroperoxides (PLOOH) and tocopherols contents
in plasma before and after the 12-week administration of 0, 6 or 12 mg astaxanthin
(Mean values and standard deviations)
0 mg 6 mg 12 mg
Parameters Mean
SD Mean SD Mean SD P
Carteonoids (pmol/ml)
Before 9 4 6 3 8 6 0·383
After 8 4 86**†† 30 109**†† 49 0·000
Before 520 82 480 88 473 75 0·398
After 529 104 489 107 484 108 0·584
Before 140 37 127 25 137 37 0·663
After 139 46 124 22 134 33 0·607
Before 283 71 266 80 259 56 0·740
After 273 66 256 73 253 71 0·802
Before 131 43 100 45 130 37 0·182
After 130 44 97 39 127 37 0·147
Before 673 50 662 70 651 68 0·742
After 679 63 667 80 660 103 0·883
Before 153 20 131 26 136 35 0·210
After 143* 19 122* 25 128* 38 0·275
Before 952 174 880 183 877 156 0·549
After 950 178 956** 188 980** 202 0·931
Non-polar carotenoidsk
Before 957 89 892 106 917 111 0·375
After 951 102 887 97 916 144 0·475
Total carotenoids
Before 1909 238 1772 261 1794 253 0·434
After 1901 237 1843* 264 1896* 320 0·872
PLOOH (pmol/ml)
Before 26·2 8·0 18·7† 5·6 18·2* 6·4 0·021
After 27·4 7·2 15·6*†† 3·4 13·9†† 4·4 0·000
Tocopherols (nmol/ml)
Before 44·7 9·3 42·8 17·6 36·9 5·6 0·328
After 45·1 10·5 39·5 8·5 38·9 10 ·7 0·326
Before 4·1 2·0 3·2 0·9 3·8 1·9 0·483
After 3·8 0·8 3·1 0·8 3·8 1·4 0·302
Total tocopherols
Before 48·8 8·7 46·0 17·4 40·6 6·6 0·314
After 48·8 10·4 42·6 8·8 42·7 11 ·0 0·304
PCOOH, phosphatidylcholine hydroperoxide; PEOOH, phosphatidylethanolamine hydroperoxide.
Mean values were significantly different in the paired t test between before and after astaxanthin administration:
*P, 0·05, **P, 0·01.
Mean values were significantly different in Dunnett’s test between the control group and one of the two astaxanthin
groups: P,0·05, ††P, 0·01.
One-way ANOVA test among groups.
§ Xanthophylls are the sum of astaxanthin, lutein, zeaxanthin and b-cryptoxanthin.
k Non-polar carotenoids are sum of a-carotene, b-carotene and lycopene.
Astaxanthin inhibits lipid peroxidation 1569
British Journal of Nutrition
. Incorporation of a carotenoid (b-carotene) into
erythrocytes after oral supplementation has been described
in some reports
. However, there has been no study evalu-
ating whether administered carotenoids other than b-carotene
are distributed to erythrocytes, except for our recent human
study of the xanthophyll lutein
. In the present study,
using the newly developed HPLC-MS/MS analysis method,
incorporation of astaxanthin into erythrocytes after oral sup-
plementation was established (Fig. 1(a)). Because both the
erythrocyte and plasma astaxanthin concentrations increased
(Tables 4 and 5), it seems likely that astaxanthin in plasma
lipoprotein particles is partly transferred into erythrocytes.
By this hypothesis, astaxanthin would be located on the
outer region of plasma lipoproteins, which would facilitate
its transfer to erythrocytes. On the other hand, the concen-
trations of endogenous antioxidants (i.e. carotenoids and
tocopherols) showed virtually no change before and after
astaxanthin supplementation (Tables 4 and 5). This is advan-
tageous for elucidating the antioxidant contribution of astax-
anthin. By the way, it was known that blood carotenoid
concentration in females is somewhat higher than that in
males. In the present study, we compared sex difference in
blood carotenoids, but no statistical differences were observed
between males and females in each carotenoid at baseline or
In the present study, to evaluate peroxidisability, we
measured the PLOOH content. Because PLOOH are the
primary oxidation products of PL, an increase in PLOOH
directly reflects in vivo oxidative stress
(1 3,22,23)
. As has been
observed, astaxanthin supplementation clearly reduced the
erythrocyte PLOOH concentration (Fig. 1(b)), indicating that
astaxanthin incorporation into erythrocytes attenuated PL per-
oxidation of erythrocyte membranes. On the other hand, the
antioxidant effect of astaxanthin seemed to be more apparent
on the erythrocyte membrane, as compared with the plasma
(Tables 4 and 5). Erythrocytes are rich in PUFA in their PL
bilayer, and contain high concentrations of molecular
oxygen and ferrous ions as constituents of oxyhaemoglobin.
The oxidation of Hb is accompanied by the formation of
superoxides, a source of reactive oxygen species. Therefore,
erythrocyte membrane PL would be more susceptible to per-
oxidation than other organelle membranes, even though
they are protected by several antioxidative systems such as
superoxide dismutase, catalase and glutathione peroxidase.
For plasma PCOOH, unexpectedly, at baseline (week 0),
groups taken 6 or 12 mg astaxanthin showed significantly
less PCOOH than the group taken 0 mg astaxanthin. Because
no differences were observed among the three groups in other
parameters (e.g. haematological and blood biochemical
values), it might be other factor(s) affecting plasma PCOOH
before the start of the study. This possibility needs further
In the present study, when comparing erythrocyte PCOOH
between the placebo and astaxanthin groups, PCOOH levels
after astaxanthin supplementation (5·2 and 6·6 pmol/ml
packed cells for 6 and 12 mg astaxanthin groups, respectively)
were somewhat lower but not statistically significant as com-
pared with those of the placebo group (9·1 pmol/ml packed
cells; P¼0·122; Table 4). In contrast, PEOOH levels after astax-
anthin supplementation (2·8 and 3·0 pmol/ml packed cells for
6 and 12 mg astaxanthin groups, respectively) were signifi-
cantly lower (P¼0·011) than those of the placebo group
(5·8 pmol/ml packed cells). PLOOH (sum of PCOOH and
PEOOH) is, therefore, considered to show significant changes
(P¼0·031) between the placebo (14·9 pmol/ml packed cells)
and astaxanthin groups (8·0 and 9·7 pmol/ml packed cells
for 6 and 12 mg astaxanthin groups, respectively). Considering
these, the antioxidant effect of astaxanthin appears likely to be
through the reduction of erythrocyte PEOOH rather than
PCOOH. This possibility needs to be clarified in future studies.
On the other hand, for the efficacy of astaxanthin, inhibitory
effects of the 6 mg astaxanthin group on PCOOH and
PEOOH were as good as or even better than those of the
12 mg astaxanthin group (Tables 4 and 5). Concentrations of
erythrocytes and plasma astaxanthin were not different
between the 6 and 12 mg astaxanthin groups, suggesting that
6 mg astaxanthin is effective enough to show antioxidative
benefit in vivo. Thus, to estimate the optimal dose of astax-
anthin, we are now conducting a human study by administer-
ing 36 mg astaxanthin to volunteers.
Among the carotenoids (xanthophylls), astaxanthin has
recently received increased scientific interest due to its
potent antioxidant activity and hence possible anti-metabolic
syndrome, anti-brain ageing and anti-atopic dermatitis
(24 26)
. We have previously found that there was a
higher accumulation of PLOOH in the erythrocytes of dementia
. Erythrocytes high in lipid hydroperoxides have been
suggested to have a decreased ability to transport oxygen to the
brain and may impair blood rheology, thus facilitating demen-
(4 8)
. In the present study, orally administered astaxanthin
was incorporated into erythrocytes, and erythrocyte PLOOH
levels decreased. On the basis of these points, it seems that simi-
lar to lutein
, astaxanthin has the potential to act as an import-
ant antioxidant in erythrocytes, and thereby may contribute to
the prevention of dementia. This possibility warrants the testing
of astaxanthin in other models of dementia with a realistic pro-
spect of its use as a human therapy.
The present study was supported in part by Grants-in-Aid for
Scientific Research (KAKENHI; 20228002) of the Japanese
Society for the Promotion of Science ( JSPS; Tokyo, Japan).
K. N. was involved in data collection, data analysis, data
interpretation, literature search and manuscript preparation.
T. K., T. M., G. C. B., F. K. and A. S. were involved in data col-
lection, data analysis and data interpretation. A. S. and T. M.
were involved in the study design, data interpretation and
review of the manuscript. None of the authors has conflicts
of interest with respect to the present study.
1. Miyazawa T, Yasuda K, Fujimoto K, et al. (1988) Presence
of phosphatidylcholine hydroperoxide in human plasma.
J Biochem 103, 744 746.
K. Nakagawa et al.1570
British Journal of Nutrition
2. Miyazawa T, Suzuki T, Fujimoto K, et al. (1992) Chemilumi-
nescent simultaneous determination of phosphatidylcholine
hydroperoxide and phosphatidylethanolamine hydroper-
oxide in the liver and brain of the rat. J Lipid Res 33, 10511059.
3. Miyazawa T, Suzuki T, Yasuda K, et al. (1992) Accumu-
lation of phospholipid hydroperoxides in red blood cell
membranes in Alzheimer disease. In Oxygen Radicals,
pp. 327330 [K Yagi, M Kondo, E Niki and T Yoshikawa,
editors]. Amsterdam: Elsevier Science Publishing.
4. Bosman GJ, Bartholomeus IG, de Man AJ, et al. (1991)
Erythrocyte membrane characteristics indicate abnormal
cellular aging in patients with Alzheimer’s disease. Neurobiol
Aging 12, 13 18.
5. Goodall HB, Reid AH, Findlay DJ, et al. (1994) Irregular
distortion of the erythrocytes (acanthocytes, spur cells) in
senile dementia. Dis Markers 12, 2341.
6. Solerte SB, Ceresini G, Ferrari E, et al. (2000) Hemorheologi-
cal changes and overproduction of cytokines from immune
cells in mild to moderate dementia of the Alzheimer’s type:
adverse effects on cerebromicrovascular system. Neurobiol
Aging 21, 271 281.
7. Ajmani RS, Metter EJ, Jaykumar R, et al. (2000) Hemo-
dynamic changes during aging associated with cerebral
blood flow and impaired cognitive function. Neurobiol
Aging 21, 257 269.
8. Mohanty JG, Eckley DM, Williamson JD, et al. (2008) Do red
blood cell-b-amyloid interactions alter oxygen delivery in
Alzheimer’s disease? Adv Exp Med Biol 614, 29 35.
9. Nakagawa K, Kiko T, Hatade K, et al. (2008) Development of
an HPLC-based assay for carotenoids in human red blood
cells: application to clinical studies. Anal Biochem 381,
10. Nakagawa K, Miyazawa T, Kiko T, et al. (2010) Effect of
carotenoids towards lipid peroxidation in vivo. J JSMUFF 6,
123130 [in Japanese].
11. Nakagawa K, Kiko T, Hatade K, et al. (2009) Antioxidant
effect of lutein towards phospholipid hydroperoxidation in
human erythrocytes. Br J Nutr 102, 1280 1284.
12. Nakagawa K, Fujimoto K & Miyazawa T (1996) b-Carotene
as a high-potency antioxidant to prevent the formation of
phospholipid hydroperoxides in red blood cells of mice.
Biochim Biophys Acta 1299, 110 116.
13. Asai A, Nakagawa K & Miyazawa T (1999) Antioxidative
effects of turmeric, rosemary and capsicum extracts on
membrane phospholipid peroxidation and liver lipid metab-
olism in mice. Biosci Biotechnol Biochem 63, 2118 2122.
14. Higuera-Ciapara I, Fe
lix-Valenzuela L & Goycoolea FM
(2006) Astaxanthin: a review of its chemistry and appli-
cations. Crit Rev Food Sci Nutr 46, 185 196.
15. Hussein G, Sankawa U, Goto H, et al. (2006) Astaxanthin,
a carotenoid with potential in human health and nutrition.
J Nat Prod 69, 443449.
16. Guerin M, Huntley ME & Olaizola M (2003) Haematococcus
astaxanthin: applications for human health and nutrition.
Trends Biotechnol 21, 210 216.
17. Satoh A, Ishikura M, Murakami N, et al. (2010) The
innovation of technology for microalgae cultivation and its
application in functional foods and the nutraceutical
industry. In Biotechnology in Functional Foods and Nutra-
ceuticals, pp. 313 329 [D Bagchi, FC Lau and DK Ghosh,
editors]. London: Taylor & Francis Group, CRC Press.
18. Ikeda S, Tohyama T, Yoshimura H, et al. (2003) Dietary
a-tocopherol decreases a-tocotrienol but not g-tocotrienol
concentration in rats. J Nutr 133, 428434.
19. Fotouhi N, Meydani M, Santos MS, et al. (1996) Carotenoid and
tocopherol concentrations in plasma, peripheral blood mono-
nuclear cells, and red blood cells after long-term b-carotene
supplementation in men. Am J Clin Nutr 63, 553558.
20. Norkus EP, Bhagavan HN & Nair PP (1990) Relationship
between individual carotenoids in plasma, platelets and
red blood cells (RBC) of adult subjects. FASEB J 4, A1774.
21. Murata T, Tamai H, Morinobu T, et al. (1992) Determination
of b-carotene in plasma, blood cells and buccal mucosa by
electrochemical detection. Lipids 27, 840843.
22. Miyazawa T (1989) Determination of phospholipid hydro-
peroxides in human blood plasma by a chemilumines-
cence-HPLC assay. Free Radic Biol Med 7, 209217.
23. Moriya K, Nakagawa K, Santa T, et al. (2001) Oxidative stress
in the absence of inflammation in a mouse model for
hepatitis C virus-associated hepatocarcinogenesis. Cancer
Res 61, 4365 4370.
24. Uchiyama A & Okada Y (2008) Clinical efficacy of astax-
anthin-containing Haematococcus pluvialis extract for the
volunteers at risk of metabolic syndrome. J Clin Biochem
Nutr 43, 38 43.
25. Satoh A, Tsuji S, Okada Y, et al. (2009) Preliminary clinical
evaluation of toxicity and efficacy of a new astaxanthin-
rich Haematococcus pluvialis extract. J Clin Biochem Nutr
44, 280 284.
26. Satoh A, Kawamura T, Horibe T, et al. (2009) Effect of the
intake of astaxanthin-containing Haematococcus pluvialis
extract on the severity, immunofunction and physiological
function in patients with atopic dermatitis. J Environ Derma-
tol Cutan Allergol 3 429438 [in Japanese].
Astaxanthin inhibits lipid peroxidation 1571
British Journal of Nutrition
... Interventional studies of the effects of supplementation with xanthophylls on lowering atherosclerosis risk * BAP, biological antioxidant potential; CRP, C-reactive protein; DBP, diastolic blood pressure; HDL, high density lipoprotein; HDL-C, high density lipoprotein-cholesterol; IMT, Intima-media thickness; ISP, isoprostane; LDL, low density lipoprotein; LDL-C, low density lipoprotein-cholesterol; MCP-1, monocyte chemoattractant protein-1; MDA, Malondialdehyde; PLOOH, Phospholipid hydroperoxide; RCT, Randomized controlled trial; SBP, systolic blood pressure; SOD, superoxide dismutase; TAC, total antioxidant capacity; TC, total cholesterol; TG, triglyceride Human intervention studies pointed out that xanthophylls reduced oxidative stress and lipid peroxidation in healthy subjects, overweight subjects, and smokers[82,[84][85][86][87]. A reduction of fatty acid oxidation was observed after the oral intake of a low dose of astaxanthin (8 mg/d) for 3 months in healthy non-smoking young men (19-33 years)[85]. ...
... A reduction of fatty acid oxidation was observed after the oral intake of a low dose of astaxanthin (8 mg/d) for 3 months in healthy non-smoking young men (19-33 years)[85]. In an RCT on healthy aging subjects (50-59 years), the administration of astaxanthin (6 or 12 mg/d) for 12 weeks significantly decreased PLOOH in erythrocytes and improved erythrocyte antioxidant status[87]. In addition, previous results showed that LDL oxidation rate decreased in response to astaxanthin supplementation (1.8, 3.6, 14.4, 21.6 mg/d) for 14 days in a dose-dependent manner in healthy young subjects(Iwamoto et al. 2000). ...
... Also, the increase in Nrf2 level increases the expressions of genes implicated in repair/removal of damaged DNA/proteins that enhance cell survival and potently protects neuronal cultures from Aβ toxicity. 85 Nakagawa et al 86 found that AST supplementation resulted in improved erythrocyte antioxidant status, reduced phospholipid hydroperoxides (PLOOH) levels, that resulted from lipid peroxidation, and reduced oxidative DNA damage in N-methyl-D-aspartate (NMDA)-induced excitotoxicity model, suggesting that astaxanthin may contribute to the prevention of dementia. Malondialdehyde (MDA) formed by free radicals during ionization events in the body, and the level of MDA in the brain increases during oxidative stress and it is used as a marker of lipid peroxidation. ...
Full-text available
Background: Astaxanthin (AST) is a second-generation antioxidant with anti-inflammatory and neuroprotective properties and could be a promising candidate for Alzheimer's disease (AD) therapy, but is shows poor oral bioavailability due to its high lipophilicity. Purpose: This study aimed to prepare and evaluate AST-loaded nanostructured lipid carriers (NLCs), for enhanced nose-to-brain drug delivery to improve its therapeutic efficacy in rat model of AD. Methods: AST-NLCs were prepared using hot high-pressure homogenization technique, and processing parameters such as total lipid-to-drug ratio, solid lipid-to-liquid lipid ratio, and concentration of surfactant were optimized. Results: The optimized AST-NLCs had a mean particle size of 142.8 ± 5.02 nm, polydispersity index of 0.247 ± 0.016, zeta potential of -32.2 ± 7.88 mV, entrapment efficiency of 94.1 ± 2.46%, drug loading of 23.5 ± 1.48%, and spherical morphology as revealed by transmission electron microscopy. Differential scanning calorimetry showed that AST was molecularly dispersed in the NLC matrix in an amorphous state, whereas Fourier transform infrared spectroscopy indicated that there is no interaction between AST and lipids. AST displayed a biphasic release pattern from NLCs; an initial burst release followed by sustained release for 24 h. AST-NLCs were stable at 4-8 ±2°C for six months. Intranasal treatment of AD-like rats with the optimized AST-NLCs significantly decreased oxidative stress, amyloidogenic pathway, neuroinflammation and apoptosis, and significantly improved the cholinergic neurotransmission compared to AST-solution. This was observed by the significant decline in the levels of malondialdehyde, nuclear factor-kappa B, amyloid beta (Aβ1‑42), caspase-3, acetylcholinesterase, and β-site amyloid precursor protein cleaving enzyme-1 expression, and significant increase in the contents of acetylcholine and glutathione after treatment with AST-NLCs. Conclusion: NLCs enhanced the intranasal delivery of AST and significantly improved its therapeutic properties.
Full-text available
Astaxanthin (AX), a lipid-soluble pigment belonging to the xanthophyll carotenoids family, has recently garnered significant attention due to its unique physical properties, biochemical attributes, and physiological effects. Originally recognized primarily for its role in imparting the characteristic red-pink color to various organisms, AX is currently experiencing a surge in interest and research. The growing body of literature in this field predominantly focuses on AXs distinctive bioactivities and properties. However, the potential of algae-derived AX as a solution to various global environmental and societal challenges that threaten life on our planet has not received extensive attention. Furthermore, the historical context and the role of AX in nature, as well as its significance in diverse cultures and traditional health practices, have not been comprehensively explored in previous works. This review article embarks on a comprehensive journey through the history leading up to the present, offering insights into the discovery of AX, its chemical and physical attributes, distribution in organisms, and biosynthesis. Additionally, it delves into the intricate realm of health benefits, biofunctional characteristics, and the current market status of AX. By encompassing these multifaceted aspects, this review aims to provide readers with a more profound understanding and a robust foundation for future scientific endeavors directed at addressing societal needs for sustainable nutritional and medicinal solutions. An updated summary of AXs health benefits, its present market status, and potential future applications are also included for a well-rounded perspective
Haematococcus pluvialis is well known for its most potent carotenoid, Astaxanthin, which protects cells from free radicals and oxidative stress due to its antioxidative nature. Due to the change in lifestyles, several chronic illnesses have recently increased uncontrollably. People are thus making an effort to control their lifestyle choices by taking natural supplements for preventative healthcare. Microalgae are useful in the biomedical and pharmaceutical sectors since they may create a wide range of bioactive chemicals. Among the wide advantages and bioactive compounds present in H. pluvialis, the king of antioxidant, astaxanthin, particularly finds application in the areas of nutrition and human health. The biomass of the H. pluvialis is rich in proteins, has low-fat content and is a good source of vital vitamins and minerals like magnesium, calcium, iron, and zinc. This microalga's bioactive chemicals have been proven to offer a wide range of health advantages, such as the treatment of cardiovascular disease, macular degeneration, anti-aging, antibacterial, anti-inflammatory, antitumor activities, etc. Moreover, astaxanthin being a naturally occurring antioxidant and non-toxic, it encourages cell-to-cell interaction, however, due to its limited bioavailability and susceptibility to environmental conditions it lacks a suitable drug delivery mechanism. As a result, astaxanthin-loaded nano carriers have been extensively studied for use in medical applications. This chapter will discuss more on the current developments and outcomes of produced formulations in the field of H. pluvialis medication delivery.Keywords Haematococcus AstaxanthinAntioxidantTherapeuticsDrug delivery
Nowadays, there is an emergent interest in new trend-driven biomolecules to improve health and wellbeing, which has become an interesting and promising field, considering their high value and biological potential. Astaxanthin is one of these promising biomolecules, with impressive high market growth, especially in the pharmaceutical and food industries. This biomolecule, obtained from natural sources (i.e., microalgae), has been reported in the literature to have several beneficial health effects due to its biological properties. These benefits seem to be mainly associated with Astaxanthin's high antioxidant and anti-inflammatory properties, which may act on several brain issues, thus attenuating symptoms. In this sense, several studies have demonstrated the impact of astaxanthin on a wide range of diseases, namely on brain disorders (such as Alzheimer's disease, Parkinson, depression, brain stroke and autism). Therefore, this review highlights its application in mental health and illness. Furthermore, a S.W.O.T. analysis was performed to display an approach from the market/commercial perspective. However, to bring the molecule to the market, there is still a need for more studies to increase deep knowledge regarding the real impact and mechanisms in the human brain.HIGHLIGHTSAstaxanthin has been mainly extracted from the algae Haematococcus pluvialisAstaxanthin, bioactive molecule with high antioxidant and anti-inflammatory propertiesAstaxanthin has an important protective effect on brain disordersAstaxanthin is highly marketable, mainly for food and pharmaceutical industries.
Full-text available
Background: Atherosclerosis can develop as a result of an increase in oxidative stress and concurrently rising levels of inflammation. Astaxanthin (AX), a red fat-soluble pigment classified as a xanthophyll, may be able to prevent the vascular damage induced by free radicals and the activation of inflammatory signaling pathways. The objective of the current study is to assess the effects of AX supplementation on cardiometabolic risk factors in individuals with coronary artery disease (CAD). Methods: This randomized double-blind placebo-controlled clinical trial was conducted among 50 CAD patients. Participants were randomly allocated into two groups to intake either AX supplements (12 mg/day) or placebo for 8 weeks. Lipid profile, glycemic parameters, anthropometric indices, body composition, Siruin1 and TNF-α levels were measured at baseline and after 8 weeks. Results: Body composition, glycemic indices, serum levels of TNF-α, Sirtuin1 did not differ substantially between the AX and placebo groups (p > 0.05). The data of AX group showed significant reduction in total cholesterol (-14.95 ± 33.57 mg/dl, p < 0.05) and LDL-C (-14.64 ± 28.27 mg/dl, p < 0.05). However, TG and HDL-C levels could not be affected through AX supplementation. Conclusion: Our results suggest that AX supplementation play a beneficial role in reducing some components of lipid profile levels. However, further clinical investigations in CAD patients are required to obtain more conclusive findings. Clinical trial registration:, identifier IRCT20201227049857N1.
Full-text available
Astaxanthin (3,3-dihydroxy-β, β-carotene-4,4-dione) is a ketocarotenoid synthesized by Haematococcus pluvialis/lacustris, Chromochloris zofingiensis, Chlorococcum, Bracteacoccus aggregatus, Coelastrella rubescence, Phaffia rhodozyma, some bacteria (Paracoccus carotinifaciens), yeasts, and lobsters, among others However, it is majorly synthesized by Haematococcus lacustris alone (about 4%). The richness of natural astaxanthin over synthetic astaxanthin has drawn the attention of industrialists to cultivate and extract it via two stage cultivation process. However, the cultivation in photobioreactors is expensive, and converting it in soluble form so that it can be easily assimilated by our digestive system requires downstream processing techniques which are not cost-effective. This has made the cost of astaxanthin expensive, prompting pharmaceutical and nutraceutical companies to switch over to synthetic astaxanthin. This review discusses the chemical character of astaxanthin, more inexpensive cultivating techniques, and its bioavailability. Additionally, the antioxidant character of this microalgal product against many diseases is discussed, which can make this natural compound an excellent drug to minimize inflammation and its consequences.
Vitamin A has an important role to play in vision, bone growth, reproduction, cell division, and cell differentiation. With the focus on Vitamin A and Carotenoids, this book includes the latest research in these areas and starts with an overview putting the compounds in context with other vitamins, supplementation and discussing the importance of beta-carotene. Details of the chemistry, structure and biochemistry of the compounds begins with nomenclature followed by information on encapsulation, thermal degradation and occurrence. Developments in analytical and bioanalytical techniques concerning these compounds in plant, milk and human tissue systems are covered in detail. Finally, the book covers the extensive functions and effects of Vitamin A on eg developmental growth, immune function, cancer risk, the brain and lungs as well as vision. Delivering high quality information, this book will be of benefit to anyone researching this area of health and nutritional science. It will bridge scientific disciplines so that the information is more meaningful and applicable to health in general. Part of a series of books, it is specifically designed for chemists, analytical scientists, forensic scientists, food scientists, dieticians and health care workers, nutritionists, toxicologists and research academics. Due to its interdisciplinary nature it could also be suitable for lecturers and teachers in food and nutritional sciences and as a college or university library reference guide.
Full-text available
Peroxidised phospholipid-mediated cytotoxity is involved in the pathophysiology of many diseases; for example, phospholipid hydroperoxides (PLOOH) are abnormally increased in erythrocytes of dementia patients. Dietary carotenoids (especially xanthophylls, polar carotenoids such as lutein) have gained attention as potent inhibitors against erythrocyte phospholipid hydroperoxidation, thereby making them plausible candidates for preventing diseases (i.e. dementia). To evaluate these points, we investigated whether orally administered lutein is distributed to human erythrocytes, and inhibits erythrocyte PLOOH formation. Six healthy subjects took one capsule of food-grade lutein (9.67 mg lutein per capsule) once per d for 4 weeks. Before and during the supplementation period, carotenoids and PLOOH in erythrocytes and plasma were determined by our developed HPLC technique. The administered lutein was incorporated into human erythrocytes, and erythrocyte PLOOH level decreased after the ingestion for 2 and 4 weeks. The antioxidative effect of lutein was confirmed on erythrocyte membranes, but not in plasma. These results suggest that lutein has the potential to act as an important antioxidant molecule in erythrocytes, and it thereby may contribute to the prevention of dementia. Therefore future biological and clinical studies will be required to evaluate the efficacy as well as safety of lutein in models of dementia with a realistic prospect of its use in human therapy.
Full-text available
Astaxanthin (Ax), a carotenoid ubiquitously distributed in microorganisms, fish, and crustaceans, has been known to be a potent antioxidant and hence exhibit various physiological effects. We attempted in these studies to evaluate clinical toxicity and efficacy of long-term administration of a new Ax product, by measuring biochemical and hematological blood parameters and by analyzing brain function (using CogHealth and P300 measures). Ax-rich Haematococcus pluvialis extracts equivalent to 4, 8, 20 mg of Ax dialcohol were administered to 73, 38, and 16 healthy adult volunteers, respectively, once daily for 4 weeks to evaluate safety. Ten subjects with age-related forgetfulness received an extract equivalent to 12 mg in a daily dosing regimen for 12 weeks to evaluate efficacy. As a result, no abnormality was observed and efficacy for age-related decline in cognitive and psychomotor functions was suggested.
Full-text available
The quantification of phospholipid hydroperoxides in biological tissues is important in order to know the degree of peroxidative damage of membrane lipids. For this purpose, optimal conditions for the chemiluminescent simultaneous assay of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) in rat liver and brain were determined. A chemiluminescence detection-high performance liquid chromatography (CL-HPLC) method that incorporates cytochrome c and luminol as a post-column hydroperoxide-specific luminescent reagent was used (Miyazawa et al. 1987. Anal. Lett. 20: 915-925; Miyazawa. 1989. Free Radical Biol. Med. 7: 209-217). An n-propylamine-bound silica column with hexane-2-propanol-methanol-water 5:7:2:1 (v/v/v/v) (flow rate 1.0 ml/min) as eluant was used to determine both PCOOH and PEOOH, which were separated from each other and from other lipids and lipid-soluble antioxidants. High reproducibility and sensitivity as low as 10 pmol hydroperoxide-O2 were observed with a mixture of 10 micrograms/ml cytochrome c and 2 micrograms/ml luminol in 50 mM borate buffer (pH 10.0, flow rate 1.1 ml/min) as luminescent reagent and a post-column mixing joint temperature of 40 degrees C. Using the established analytical conditions, it was confirmed that both PCOOH (1324 +/- 122 pmol/g liver, 114 +/- 18 pmol/g brain, mean +/- SD) and PEOOH (728 +/- 89 pmol/g liver, 349 +/- 60 pmol/g brain, mean +/- SD) are present in the liver and brain of Sprague-Dawley rats bred on a slightly modified AIN-76A semisynthetic diet for 3 months. The phospholipid hydroperoxide content in the rat liver was shown to be affected by dietary oils, but not significantly affected in the brain.
We previously showed that alpha- and gamma-tocotrienols accumulate in adipose tissue and skin but not in plasma or other tissues of rats fed a tocotrienol-rich fraction extracted from palm oil containing alpha-tocopherol and alpha- and gamma-tocotrienols. To clarify the nature of tocotrienol metabolism, we studied the distribution of alpha- or gamma-tocotrienol in rats fed alpha- or gamma-tocotrienol without alpha-tocopherol, and the effect of alpha-tocopherol on their distribution. Wistar rats (4-wk-old) were fed a diet with 50 mg alpha-tocotrienol/kg alone or with 50 mg alpha-tocopherol/kg in expt. 1, and a diet with 50 mg gamma-tocotrienol/kg alone or with 50 mg a-tocopherol/kg in expt. 2, for 8 wk. alpha-Tocotrienol was detected in various tissues and plasma of the rats fed alpha-tocotrienol alone, and the alpha-tocotrienol concentrations in those tissues and plasma decreased (P < 0.05) by the dietary alpha-tocopherol in the rats fed alpha-tocotrienol with alpha-tocopherol. However, gamma-tocotrienol preferentially accumulated in the adipose tissue and skin of the rats fed gamma-tocotrienol alone, and the dietary alpha-tocopherol failed either to decrease (P greater than or equal to 0.05) gamma-tocotrienol concentrations in the adipose tissue and skin or to increase (P greater than or equal to 0.05) in the urinary excretion of 2,7,8-trimethyl-2(2'-carboxym ethyl)-6-hydroxycroman, a metabolite of gamma-tocotrienol, in the rats fed gamma-tocotrienol with alpha-tocopherol. These data suggest that alpha-tocopherol enhances the alpha-tocotrienol metabolism but not the gamma-tocotrienol metabolism in rats.
Peroxidized phospholipid-mediated cytotoxicity is involved in the pathophysiology of many diseases; for example, there is an abnormal increase of phospholipid hydroperoxides in red blood cells (RBCs) of dementia patients. Dietary carotenoids have gained attention as potent inhibitors of RBC phospholipid hydroperoxidation, thereby making them plausible candidates for preventing disease. However, the occurrence of carotenoids in human RBCs is still unclear. This is in contradistinction to plasma carotenoids, which have been investigated thoroughly for analytical methods as well as biological significance. In this study, we developed a method to analyze RBC carotenoids using high-performance liquid chromatography (HPLC) coupled with ultraviolet (UV) diode array detection (DAD) and atmospheric pressure chemical ionization (APCI) mass spectrometry (MS). Under optimized conditions that included extraction, separation, and detection procedures, six carotenoids (lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, beta-carotene, and lycopene) were separated, detected by DAD, and concurrently identified based on APCI/MS and UV spectra profiles when an extract from human RBCs was subjected to HPLC-DAD-APCI/MS. The amounts of carotenoids varied markedly (1.3-70.2 nmol/L packed cells), and polar oxygenated carotenoids (xanthophylls) were predominant in RBCs. The HPLC-DAD-APCI/MS method would be a useful tool for clinical studies for evaluating the bioavailability of RBC carotenoids.
The beta-carotene concentrations in plasma, blood cells and buccal mucosal cells were determined by high-performance liquid chromatography with electrochemical detection. This method was 1,000 times more sensitive than the conventional spectrophotometric method. Polymorphonuclear cells and red blood cells had lower beta-carotene levels than the other cells. After oral administration of 580 mg/day of all-trans beta-carotene to human male volunteers for a week, the beta-carotene concentrations in all cell types increased at least several times above the original levels.