Modern Scientific Press

Copyright © 2012 by Modern Scientific Press Company, Florida, USA

International Journal of Food Nutrition and Safety, 2012, 1(2): 75-98

International Journal of Food Nutrition and Safety

Journal homepage: www.ModernScientificPress.com/Journals/IJFNS.aspx

ISSN: 2165-896X Florida, USA

Review

Analytical Methods and Bioactivities of Lutein

Jing Zhou 1, Sha Li 1, Shan Wu 1, Xiang-Rong Xu 2, Gui-Fang Deng 1, Fang Li 1, Hua-Bin Li 1, *

1 Guangdong Provincial Key Laboratory of Food, Nutrition and Health, School of Public Health, Sun

Yat-Sen University, Guangzhou 510080, China 2 Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of

Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China

* Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.:

+86-20-87332391; Fax: +86-20-87330446.

Article history: Received 18 April 2012, Received in revised form 12 May 2012, Accepted 14 May

2012, Published 15 May 2012.

Abstract: Lutein is a fat-soluble carotenoid, and could be found in a wide range of natural

sources, such as algae, tea, herbs, fruits and vegetables. A variety of separation and

analytical methods have been developed for the determination of lutein, including solvent

extraction, supercritical fluid extraction, and high performance liquid chromatography.

Lutein has showed many bioactivities, such as antioxidant and free radicals scavenging,

eyes health protection, antitumor, anti-inflammation, and protective effects on diabetes and

cardiovascular diseases. The objective of this review is to provide an up-to-date coverage

of lutein with reference to analytical methods, natural sources and bioactivities, and special

attention is paid to its bioactivities.

Keywords: lutein; natural source; bioactivity; antioxidant; anti-inflammation; anticancer;

separation; determination.

1. Introduction

Lutein is classified into the carotenoids family and belongs to the group of xanthophylls, the

oxygenated derivatives of carotenes. It is fat-soluble and sensitive to light, heat, oxygen and acids, with

the ionone ring systems being substituted at both the 3 and 3′carbon. Its chemical structure is shown in

Fig. 1. Lutein can not be synthesized by human so it must be obtained through dietary sources. A great

amount of studies have found that lutein has a range of biological activities, for instance antioxidation,

Int. J. Food Nutr. Saf. 2012, 1(2): 75-98


76

preventing cataract and age-related macular degeneration, antitumor, anti-inflammation, and protective

effects on diabetes and cardiovascular diseases (Astner et al., 2007; Goodman et al., 2007; Moeller et

al., 2008; Muriach et al., 2008; Rafi and Shafaie, 2007; SanGiovanni et al., 2007; Sato et al., 2011).

This review provides an up-to-date coverage of lutein with reference to natural source, analytical

method and bioactivities, and special attention is paid to its bioactivities.

HO

OH

Figure 1. The chemical structure of lutein

2. Separation and Analytical Methods of Lutein

Several methods, such as solvent extraction, supercritical fluid extraction and high performance

liquid chromatography, have been developed for the extraction, separation and determination of lutein.

Solvent extraction is a traditional method for extraction of lutein from various materials, such as corn,

carrots, microalgae, marigold flower, and plasma. The solvent extraction methods of lutein involve one

or more organic solvents, including hexane, tetrahydrofuran, methanol, acetone, acetonitrile, ethanol,

and petroleum ether (Azevedo-Meleiro and Rodriguez-Amaya, 2007; Hojnik et al., 2008; Kim et al.,

2007). A byproduct of corn and soybean industries, ethyl lactate, was found to be a potential solvent

for extraction of lutein from white corn and carrots prepared as dried powders (Ishida and Chapman,

2009). In a study on the solvent extraction kinetics behavior of lutein from marigold flower petals, the

result showed that the intra-particle diffusion was the rate-governing step of the extraction process, and

the isolation of free lutein could be achieved by a singlet-step procedure, in which the optimal

parameters includes as follow: hexane as organic solvent, temperature 40 �, plant particle size < 0.315

mm, Rs (solvent/material) ＝5 L/kg, Ra (alkali solution/material) ＝ 3.75 L/kg and concentration of

alkali solution of potassium hydroxide in ethanol of 5% (w/v) (Hojnik et al., 2008). The most effective

solvent for extraction of lutein from marigold flower was reported to be methanol, in comparison with

utilizing hexane, acetone, petroleum ether, using astaxanthin as the internal standard (Bhattacharyya et

al., 2008). However, with regard to industrial applications in extraction of lutein, most of the

traditional organic solvents can not be helpful due to their toxic influence on human heath.

Supercritical fluid extraction is an alternative extraction method to conventional solvent

extraction method, and could be used for extraction of lutein (Gomez-Prieto et al., 2007; Machmudah

et al., 2008; Macias-Sanchez et al., 2010). In the field of supercritical fluid extraction techniques, the



77

prominent mobile phase is carbon dioxide in the form of supercritical fluid, which is non-toxic and low

environmental impact as well as easily separated from extracts by simply reducing the pressure. The

supercritical fluid extraction is sensitive to temperature and pressure conditions (Macias-Sanchezet al.,

2010), and increasing temperature could obtain higher extraction yields for lutein and cis-lutein from

Mentha spicata L. (Gomez-Prietoet al., 2007). The optimal conditions for supercritical fluid extraction

of lutein and zeaxanthin from a popular plant consumed as a nutritious food and traditional medicine in

China, daylily (Hemerocallis disticha) flowers were 80 � and 600 bar (Hsu et al., 2011). Except of the

appropriate increase of temperature and pressure, the extraction efficiency for lutein in the supercritical

fluid extraction-carbon dioxide process could be enhanced using ethanol and acetone called entrainer

or modifier (Kitada et al., 2009; Shi et al., 2010). Besides its strong power in extracting lutein from

various samples, the supercritical fluid extraction-carbon dioxide process can also used to re-crystallize

lutein from organic solvent solutions, and the best process conditions for extracting lutein from ethyl

acetate solutions has been studied, which came out to be pressure above 8 MPa, initial concentration

below 700 ppm and CO2 mol ratio above 96.4% (Miguel et al., 2008). Nevertheless, it is reported that

supercritical carbon dioxide was less effective than subcritical propane in the extraction of lutein

diesters from marigold (Tagetes erecta) flower petals (Skerget et al., 2010). Yet when compared to the

organic solvent extraction method, supercritical fluid extraction-carbon dioxide extraction achieved a

result of much higher content of lutein (Shi et al., 2010). In addition, other extraction methods have

been developed for extracting lutein or lutein esters, such as supercritical fluid extraction-carbon

dioxide combined with ultrasound (Gao et al., 2009), ultrasound-assisted solvent extraction (Yue et al.,

2006), and enzymatic treatment extraction (Li and Han, 2009).

A range of chromatographic techniques have been exploited to analyze lutein and its isomers as

well as associated pigments both quantitatively and qualitatively in many species like algae, higher

plants and in biological samples (e.g. plasma, milk). HPLC is widely used for the determination of

lutein and its derivatives (Al-Duais et al., 2009; Chu et al., 2010; Huang et al., 2010; Li et al., 2007;

Liu et al., 2007; Mamatha et al., 2011), and some examples are listed in Table 1.

In terms of the choice of the stationary phase, C30 and C18 columns with fillers (e.g. YMC,

polymeric silica) were usually taken into account, with C16-Amide column being selected sometimes,

in the size of 250 × 4.6 mm i.d. or 150 × 4.6 mm i.d. or 55 × 4 mm. Nonetheless, Kimura et al. (2007)

has compared the concentrations of lutein from sweet potato, cassava and maize samples obtained with

C30 and C18 columns, and found that the values were very similar. The results of the study utilizing

C16-Amide column supported the idea that C16-Amide column is more suitable for separating non-polar

hydrocarbon carotenes rather than the polar lutein. Some studies reported to use a pre-column or called

as a guard column, ranging from 10 × 4.6 mm i.d to 150 × 4.6 mm i.d. The particle size in the column



78

ranged from 3 ~ 5 µm. On the chromatographic conditions, gradient elution procedure was more

frequently manipulated than its isocratic counterpart. There is a large variety in solvent composition,

but the types of them vary only a little. The solvent system often involved with solvent including

methanol, ter-butyl methyl ether, acetonitrile, ethyl acetate, methylene chloride and H2O, in the form

of unitary, binary or ternary solvent system. The column temperature ranged from 20 � to 30 �, with

flow rate was controlled in the range of 0.2 ~ 1.3 mL/min. The injection volume was either 10 µm or

20 µm. Several detection techniques have been used for HPLC analysis of lutein. Due to its time-

saving, high accuracy and reproductivity, photodiode array detector are usually coupled with HPLC to

analyze lutein and associated pigments, monitoring at wavelengths from 420 to 480 nm. In addition,

HPLC coupled with ultraviolet-visible and mass spectrometric detection seems to be a promising

technique for identifying and quantifying lutein in natural products. The evidences came from a

research on structural and geometrical isomers of carotenoids in mature grapes (Crupi et al., 2010), in

which simultaneous determination of four isomers of lutein and two β-carotenes (trans- and cis-

isomers) as well as many other phytochemicals that could not be identified and differentiated in many

studies, was realized.

Table1. Determination of lutein and its derivates by HPLC

Sample Column Condition Detection Reference

caper leaves

(Capparis

spinosa)

Zorbax C18

(250 × 4.6 mm) gradient procedure: A: acetonitrile/

water (90/10 , v/v), B: ethyl acetate;

flow rate, 1 mL/min; injection

volume, 10 µL

PAD,

440 nm

Tlili et al.,

2009

daylily

flowers

(Hemero-

callis

disticha)

RP-YMC C30

column (250 ×

4.6 mm, 5 µm)

isocratic procedure: methanol/

methyl-tert-butyl ether (86/14, v/v);

flow rate, 1 mL/min; injection

volumn, 20 µL

multi-

wavelength

detector,

450 nm

Hsu et al.,

2011

Dunaliella

salina RP-YMC C30

column

(250 × 4.6 mm,

5 µm)

isocratic procedure: mixing

methanol-acetonitrile-water (84/14/2,

v/v/v) and methylene chloride (75/25,

v/v); flow rate, 1 mL/min; injection

volume, 20 µL

PAD, 450

nm

Hu et al.,

2008

fruit, macula

of egg,

vegetables

Purospher Star

(55 ×4 mm, 3

µm)

isocratic procedure:100﹪pure ACN;

column temperature, 25 ± 1℃;

flow rate, 0.4 mL/min

PAD, 448,

450 nm

Velpandian

et al., 2010

grape

(Negro-

amaro)

RP-YMC C30 (

250 × 3 mm, 5

µm); a pre-

gradient procedure: A: 0.05﹪TEA in

H2O, B: 0.05﹪TEA in methanol,

C: 0.05﹪TEA in ter-butyl methyl

UV–vis

detector,

447 nm;

Crupi et al.,

2010



79

column, YMC

C30 (20 × 3

mm, 5 µm)

ether; flow rate, 0.2 mL/min;

injection volumn, 10 µL

ESI-MS,

100 ~ 1200

m/z

Makino

(Gynos-

temma

pentaphy-

llum)

YMC C18

column (250 ×

4.6 mm, 5 µm)

gradient procedure: A: methanol/

acetonitrile/ water (84/14/2, v/v/v),

B: methylene chloride;

flow rate, 1.0 mL/minute;

column temperature, 25�

PAD, 450

nm;

ESI/APCI-

MS

Tsai et al.,

2010

leafy

vegetables

RP-Phenom-

enex C18 （ 250

× 4.60 mm, 5

µm)

gradient procedure: A: acetonitrile

and 0.05﹪TEA, B: methanol/ethyl

acetate (1/1, v/v); column

temperature, 25℃; flow rate, 1.0

mL/min; injection volumn,10 µL

UV-Vis

detector,

450 nm

Belanger et

al., 2010

Naranjilla or

Lulo

(Solanum

quitoense

Lam.)

RP-YMC

C30 (250 × 4.6

mm, 5 µm)

gradient procedure: A: water, B:

methanol, C: methyl tert-butyl ether;

column temperature, 25 �; flow rate,

1 mL/min; injection volume, 20 µL

PAD, 450

nm

Gancel et

al., 2008

Phyto-

plankton

C16-Amide

column (150

mm × 4.6 mm,

5 µm), a guard

column of the

same material

gradient procedure, A: methanol,

B: acetonitrile, C: 0.5 M ammonium

acetate; column temperature, 30 �

PAD, 440

and 450 nm

Jayaraman

et al., 2011

red blood

cells

YMC C30

(250 mm × 4.6

mm, 5 µm)


methyl tert-butyl ether/water

(83/15/2, v/v/v) containing 3.9

mmol/L ammonium acetate,

B: methanol/methyl tert-butyl

ether/water (8 /90 /2, v/v/v)

containing 2.6 mmol/L ammonium

acetate; column temperature: 20 �;

flow rate, 1 mL/min

PAD, 463

nm;

APCI-MS

scanning

range, 200

~ 800 m/z

Nakagawa

et al., 2008

pumpkin

(Curcurbita

moschata),

orange flesh

RP-YMC C30

(250 mm × 4.6

mm, 5 µm)


methyl tert-butyl ether/water

(81/15/4, v/v/v), B: methanol/

methyl-tert-butyl ether/water (4/92/4,

v/v/v); flow rate, 0.42 mL/min;

injection volumn, 20 µL

PAD,

450 nm

Shi et al.,

2010

potato YMC C30

(250 mm × 4.6

gradient procedure: methanol/methyl-

tert-butyl ether; flow rate,

PAD,

450 nm

Burgos et

al., 2009



80

Ultra-performance liquid chromatography (UPLC), usually coupled with photodiode array

detection or tandem mass spectrometric detection, are also used for the analysis of lutein and its

derivatives (Dugo et al., 2008; Gentili and Caretti, 2011; Granado-Lorencio et al., 2010; Karlina et al.,

2008; Pham and Hatcher, 2011; Rivera et al., 2011). A comparison between a ultra-performance liquid

chromatography and a conventional HPLC method came to a conclusion that performance of the

former transcended that of the latter in separating lutein from forage (Chauveau-Duriot et al., 2010).

Aside from chromatographic method, there also were other methods applied in the analyses of lutein

and its derivatives, such as ultraviolet-visible spectrophotometry, near infrared reflectance

spectroscopy, and fluorescence quenching method (Belefant-Miller and Grace, 2010; Bonierbale et al.,

2009; Chen et al., 2009; Davey et al., 2009; Kupper et al., 2007).

3. Natural Sources of Lutein

The most well-known natural source of lutein is marigolden flower and this pigment has been

reported to be present in a wide rang of natural sources, like algae, tea, herbs, fruits and vegetables

(Chandrika et al., 2011; Daly et al., 2010; Guil-Guerrero and Rebolloso-Fuentes, 2008; Kobori and

Arnaya, 2008; Ren and Zhang, 2008; Wang et al., 2010; Yahia et al., 2011). It has been reported that

green leafy vegetables were the major contributors of dietary lutein (Perry et al., 2009). Many

researchers have tried to established databases of individual carotenoids including lutein contents in

vegetables and fruits, and these data would be helpful for estimating the dietary intake of lutein and

provide a base for epidemiological studies on the relation between lutein intake and the risk of chronic

diseases (Aizawa and Inakuma, 2007; Chandrika et al., 2010; Dias et al., 2009; Mamathaet al., 2011;

Murillo et al., 2010). Some natural sources of lutein are given in Table 2. It is believed that the quality

and quantity of lutein in various species is likely to be affected by plant variety, portions, growing

environment and seasonality, storage conditions, maturity stages and processing methods (Hidalgo and

mm, 3 µm) 0.8 mL/min; injection volumn, 10 µL

spiny sea-

star

(Marthas-

terias

glacialis)

Luna RP-C18

(250 × 4.6 mm,

5 µm)

isocratic procedure: acetonitrile;

flow rate, 0.5 mL/min; injection

volume, 40 µL

PAD, 300

~ 650 nm;

APCI- MS,

200 ~ 1500

m/z

Ferreres et

al., 2010

Vitaceae

(Cyphos-

temma

digitatum)

leaves

RP-C30 (250 ×

4.6 mm, 5 µm);

C30 guard

column (10 ×

4.6 mm, 5µm)

gradient procedure: TBME/MeOH;

column temperature, 17 ± 1 �; flow

rate, 1.3 mL/min

PAD, 470

nm

Al-Duais et

al., 2009



81

Brandolini, 2008; Holasova et al., 2009; Lozano et al., 2011; Pinto et al., 2011; Provesi et al., 2011;

Wang et al., 2008; Yang et al., 2010; Zanatta and Mercadante, 2007; Zhang et al., 2009).

4. Bioactivities of Lutein

4.1. Antioxidant and Free Radicals Scavenging Activities

Many experiments have been done in the area of antioxidative assays of lutein and the pigment

has been identified as an important protective agent offering resistance to oxidative stress by

scavenging free radicals, inhibiting cell membrane damage and lipid peroxidation, etc. Several in vitro

researches confirmed that lutein was an effective scavenger of hydrogen peroxide (H2O2), superoxide

anion and free radicals, such as hydroxyl free radical, lipid free radical, 1,1-diphenyl-2-picrylhydrazyl

(DPPH) radical, and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS•+), as

well as an active inhibitor of lipid peroxidation and iron-mediated Fenton reaction (Bhattacharyya et

al., 2010; Hsuet al., 2011; Kumar et al., 2010). Another in vitro experimental study indicated that the

free radical scavenging ability of lutein exhibited in a concentration-dependent manner, and the

effective inhibition concentration (IC50) values were 7.54, 3.54, 7.70, 7.75 mg/mL for the 2,2-

diphenyl-1-picrylhydrazyl radicals, 2,2-azobis-3-ethylbenzthiazoline-6-sulphonic acid radicals,

inhibition of RBC hemolysis induced by peroxide, and inhibition of lipid peroxidation assays,

respectively (Fu et al., 2011). Lutein was also able to capture peroxynitrite and nitrogen dioxide

radicals from their molecules and so it may be reasonable to presume that this pigment could protect

DNA, protein and low density lipoprotein since peroxynitrite is known to be capable of inducing these

substances to alter (Tsuboi et al., 2010). This is in accordance with the results of a study, which

showed that lutein protected against DNA damage in SK-N-SH human neuroblastoma cells induced by

reactive nitrogen species, including S-nitrosoglutathione monoethyl ester, Na2N2O3 and 3-morpholino-

sydnonimine (Santocono et al., 2007). An in vitro study reported that lutein protected HT-29 cells from

deoxynivalenol-induced oxidative stress through suppressing expression of inflammatory genes, NF-

κB and cyclo-oxygenase-2, and cytotoxicity by maintaining cell viability and reducing lactate

dehydrogenase leakage (Krishnaswamy et al., 2010).

The efficacy of quenching free radicals of lutein was compared with other natural pigments or

products. The results of a fundamental study about the free radical scavenging activity of lutein, β-

carotene and violaxanthin purified from water spinach revealed that the free radicals scavenging ability

followed the order, considering the effective inhibition concentration (IC50) values: β-carotene > lutein

> violaxanthin for the 1,1-diphenyl-2-picrylhydrazyl radical assay; violaxanthin > lutein > β-carotene

for the 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation assay; violaxanthin > β-

carotene > lutein for the inhibition of RBC hemolysis induced by hydrogen peroxide and inhibition of



82

lipid peroxidation assays (Fu et al., 2011). When comparisons of in vitro antioxidant capacity among

the four commercially available natural products lutein, olive leaf extract, sesamol and ellagic acid

were carried out, antioxidant efficacy followed the order: ellagic acid > sesamol > olive leaf extract >

lutein for all the antioxidant test methods, 1,1-Diphenyl-2-picrylhydrazyl, 2,2-Azinobis(3-ethylbenzo-

thiazoline-6-sulfonic acid) radical cation, ferric reducing antioxidant capacity, oxygen reducing

antioxidant capacity and β-carotene-linoleic acid assays (Hayes et al., 2011).

Table 2. Some natural sources of lutein

Source Lutein (%) Method Reference

camu-camu fruit (Myrciaria

dubia)

peel

160.5 ± 93.1a µg/100 g FW

601.9 ± 75.6a µg/100 g FW

HPLC-PAD Zanatta and

Mercadante,

2007

C. maxima × C. moschata hybrid

‘Tetsukabuto’

C. pepo ‘Mogango’ flesh

56.6 ± 9.7 µg/g FW

9.8 ± 2.9 µg/g FW

HPLC-PAD Azevedo-Meleiro

and Rodriguez-

Amaya, 2007

caper (Capparis Spinosa)

leaves

buds

flowers

2346.3 ± 1292.9 µg/g FW

672.9 ± 402.7 µg/g FW

214 ± 117.9 µg/g FW

HPLC-PAD Tlili et al., 2009

daylily (Hemerocallis disticha)

flowers

245.61 ± 5.12 µg/g DW HPLC Hsuet al., 2011

jackfruit (Artocarpus

heterophyllus) pulp

37.02a µg/100 g FW HPLC-PDA de Faria et al.,

2009

jambolao fruits (Syzygium

cumini) pulp and peel

39.0 ± 2.2a µg/100 g FW HPLC-PAD-

MS/MS

Faria et al., 2011

Merlot grape berries (Vitis

vinifera cv. Merlot)

1.51 µg/g FW

1.73 µg/g FW

HPLC-PAD Kamffer et al.,

2010

spinach (Spinacia oleraceea)

variety Leopold leopard leaves

52.2 ± 0.9 mg/kg FW HPLC Bunea et al.,

2008

Violadeclinata herba aerial parts 896 µg/100g FW HPLC Toiu et al., 2009

Note: a All-trans-lutein; FW = fresh weight; DW = dried weight.

The role of lutein as an antioxidant was reinforced by the results of several in vivo tests (Sato et

al., 2011; Serpeloni et al., 2010; Sindhu et al., 2010). The results of one in vivo experiment

demonstrated that lutein exerted a hepatoprotective role, which was believed to be related to lutein’s

capacity to scavenging reactive oxygen radicals, in rats treated with paracetamol, carbon tetrachloride

or ethanol (Sindhuet al., 2010). Another in vivo study using ischemia-reperfusion model rats found that

lutein had a potent protective effect against oxidative injury (Sato et al., 2011). In addition, this



83

carotenoid was found to be capable of reducing the formation of crosslink and chromosome instability

induced by cisplatin as well as increasing glutathione levels in mice (Serpeloni et al., 2010). An

investigation on the individual carotenoid content by HPLC revealed that lutein was relatively lacking

in human skin as compared to plasma (Scarmo et al., 2010). Two randomized, double-blind, placebol-

controlled clinical trials discovered that lutein may improve skin health upon its increasing skin

hydration, lipid content and elasticity as well as decreasing skin lipid peroxidation activity (Maci 2007;

Palombo et al., 2007). However, it should be pointed out that extra supplementation with lutein or

compounds rich-in lutein may not significantly affect body antioxidant status or oxidative stress in

normal healthy subjects when sufficient amounts of antioxidants already exist. This has been verified

by an investigation on whether supplementation of lutein and/or green tea can alter overall antioxidant

status/oxidative stress in a well nourished population aged 50 – 70 years (Li et al., 2010).

4.2. Activities on Eyes Health

Potential benefits on eyes health of lutein have received extensive attention since this pigment

was reported as a component of the human retina together with zeaxanthin (Bone et al., 1985). A

number of in vitro tests, animals models, clinical trials or epidemiological studies have been conducted

to evaluate the potential protective effect of lutein against retinal neural damage, age-related macular

degeneration, cataract and so forth (Cho et al., 2008; Hu and Xu, 2008; Johnson et al., 2008; Li and

Lo, 2010; Sasaki et al., 2009). The most well-known activity of lutein has been implicated in the

protection of age-related macular degeneration (AMD). High levels of lutein/zeaxanthin intake from

diet were observed to relate to low risk of generating age-related macular degeneration, geographic

atrophy, and large or extensive intermediate drusen (SanGiovanni et al., 2007). Lutein supplementation

may increase the macular pigment optical density, thus benefit visual function in patients with age-

related macular degeneration (Weigert et al., 2011). However, it is reported that the combination of

lutein with vitamins and minerals at 6 mg dose did not benefit patients with age-related maculopathy

and atrophic age-related macular degeneration participants (Bartlett and Eperjesi, 2007). In addition, a

study showed that associations between lutein/zeaxanthin intake and the risk of self-reported early

AMD did not exist according to the data from a prospective follow-up investigation of 71, 494 women

and 41, 564 men (Choet al., 2008). The same conclusion was reached by a systematic review and

meta-analysis (Chong et al., 2007), in which nine prospective cohort studies and three randomized

clinical trials were finally included, using standardized criteria. The results of the meta-analysis

supported that evidence to the protective role which dietary lutein or other antioxidants like vitamin A

and vitamin C, including their supplements, play against early AMD was insufficient.



84

Evidence to the neuroprotective role of lutein has been supported by an in vitro model of

hypoxia and oxidative stress, results of which revealed that retinal ganglion cells RGC-5 were

protected by lutein from chemical hypoxia induced by cobalt chloride and oxidative stress induced by

hydrogen peroxide (Li and Lo, 2010). The role of lutein as a neuroprotectant has been reinforced by

data from a model of acute retinal ischemia/reperfusion (Li et al., 2009). Lutein might also be applied

to prevent after-cataract, based on the conclusion of an in vitro experiment using bovine lens epithelial

cells model, in which lutein could inhibit the proliferation and migration of cells at concentration of ≥

1 µmol/L (Hu and Xu, 2008). The risk of nuclear cataract in older women might be decreased with

diets rich in lutein and zeaxanthin (Moelleret al., 2008).

Filtering blue-light and quenching reactive oxygen species have been proposed to be major

mechanisms for how lutein performs its biological function on ocular health (Krinsky et al., 2003;

Snodderly, 1995). The former has been authenticated indirectly by an animal experiment (Barker et al.,

2011), in result of the fovea of rhesus monkey eyes being protected against blue light-induced damage

by lutein supplementation after long-term xanthophyll deficiency. In support of the reducing oxidation

mechanism, Li et al. (2010) reported that singlet oxygen was generated in human retinal pigment

epithelium/choroid but not retina, and lutein along with other macular pigments located in

photoreceptor outer segments might be able to directly quench them. Pintea et al. (2011) also agreed

with the ideas that lutein and zeaxanthin supplementation could protect human retinal pigment

epithelium cells D407 RPE by helping them against hydrogen peroxide induced oxidative stress.

4.3. Antitumor Activities

Many studies related to the risk for cancer and carcinogenesis have been performed to probe

into the role which lutein play in protecting from several cancers. An in vitro test found that lutein

showed inhibiting action to human hepatoma cell Hep3B in G (0)/G (1) phase of cell cycle and

inducing Hep3B cells necrosis or apoptosis (Tsai et al., 2010). Additionally, a Wistar rat model

clarified lutein as a suppressing agent during diethyllnitrosamine-induced hepatocarcinogenesis

(Moreno et al., 2007). The extract almost completely composed of lutein from Chlorella vulgaris has

been found to exhibit antiproliferative activity on human colon cancer cells HCT116 in a dose-

dependent manner, with an effective inhibition concentration (IC50) value of 40.31 ± 4.43 µg/mL (Cha

et al., 2008). Moreover, a dimethylhydrazine-treated rat model demonstrated the chemo-protective

effect of lutein against colon carcinoma through regulating the proliferative activity of K-ras, protein

kinase B, and β-catenin proteins in tumors (Reynoso-Camacho et al., 2011). The ultraviolet B-

irradiated Skh-1 hairless mice model confirmed that dietary supplementation with lutein/zeaxanthin (in

a ratio of 20:1) reduced photoaging and photocarcinogenesis in mice skin (Astneret al., 2007). The



85

results of quite a few investigations or clinical trials have suggested that a diet rich in lutein along with

other carotenoids may reduce the risk of several kinds of cancers, such as prostate cancer, epithelial

ovarian cancer, endometrial cancer, and cervical or breast cancer (Beydoun et al., 2011; Ghosh et al.,

2008; Mignone et al., 2009; Pelucchi et al., 2008; Zhang et al., 2007a & b). However, a case-control

study reported that lutein/zeaxanthin did not appear to be protective agents for lung cancer(Comstock

et al., 2008). In addition, a cohort study suggested that lung cancer prevention should avoid long-term

use of individual lutein, β-carotene and retinol supplements, particularly among smokers (Satia et al.,

2009).

4.4. Other Bioactivities

Additional biological functions of lutein have been demonstrated, including protective effects

on diabetes and related diseases, anti-inflammatory and antibiosis, and protective effects on

cardiovascular diseases, which are summarized in Tables 3-6, except the aforementioned.

Table 3. Effects of lutein on diabetes and associated diseases

Model Function Reference

diabetic Wistar rats normalize the diabetes-induced biochemical,

histological, and functional modifications in the retina

Arnal et al., 2009

streptozotocin-

induced diabetic rat

attenuate the oxidative changes of the diabetic cerebral

cortex

Arnal et al., 2010

streptozotocin-

induced diabetic rat

prevent the development and progression of cataracts Arnal et al., 2009

lymphocytes from

diabetic rats

prevent oxidative stress, suppressed the NF-κ B

activation induced by high glucose

Muriachet al.,

2008

human-derived retinal

pigment epithelial

ARPE-19 cells

suppress cell proliferation and mRNA upregulation of

vascular endothelial growth factor and matrix

metalloproteinases-2 induced by exogenous advanced

glycation endproducts

Sun et al., 2011

patients with

nonproliferative

diabetic retinopathy

improve visual acuity, increase contrast sensitivity,

decrease fovea thickness

Hu et al., 2011

streptozotocin-

induced diabetes

C57BL/6 mice

prevent local oxidative stress in the diabetic retina,

prevent decrease in the thickness of the inner

plexiform and nuclear layers and ganglion cell number

Sasaki et al., 2010



86

Table 4. Anti-inflammatory effects of lutein

Model Possible Mechanism Reference

LPS-stimulated

mouse macrophage

cell line RAW 264.7

decrease the expression of the inducible nitric oxide

synthase at both the mRNA and protein levels

Rafi and Shafaie,

2007

lipopolysaccharide

(LPS)-stimulated

macrophages

inhibit the redox-sensitive activation of the NF-κ B-

inducing kinase and phosphatidylinositol 3-kinase as

well as oxidative inactivation of PTEN

Kim et al., 2008

specific isoforms of

mammalian DNA

polymerases

inhibit mammalian DNA polymerases β and λ Horie et al., 2010

gastric epithelial

AGS cells

inhibit hydrogen peroxide-induced activation of NF-κB

and IL-8 expression

Kim et al., 2011

Table 5. Antibiosis effects of lutein

Model Possible Mechanism Reference

stable HBV-producing

human hepatoblastoma

HepG2 2.2.15 cells

suppress the secretion of HBsAg, inhibit the

activity of HBV full-length promoter (Fp),

inhibited HBV transcription

Pang et al., 2010

a cohort of 279 Hawaiian

residents

reduce the risk of anal human papillomavirus

infections

Shvetsov et al.,

2010

Table 6. Protective effects of lutein on cardiovascular disease

Model Function Reference

cohort of 63, 257

Chinese men and women

decreased risk of acute myocardial infarction (Koh et al., 2011)

Aorta of guinea pigs prevent cholesterol accumulation, decrease

oxidized LDL and inflammatory cytokines

(Kim et al., 2011)

5. Conclusions and Prospects

Lutein could be extracted from various natural sources or biological samples using solvent

extraction, supercritical fluid extraction, ultrasound-assisted extraction, and enzymatic treatment

extraction. Lutein is usually determined by HPLC or UPLC with PAD or mass spectrometric detection.

As a result of the attempt to uncovering possible contributions to human health of lutein, the pigment

has been reported to be associated with antioxidant and free radicals scavenging, eyes health protection,



87

antitumor, anti-inflammatory, and antibiosis activities, and protective effects on diabetes along with

associated diseases. In the future, more widely pharmacological studies should be carried out to

explore new pharmacodynamic effects. Furthermore, the action mechanism of lutein should be

investigated thoroughly. In addition, more attentions should be paid to develop methods for

simultaneous determination of lutein and the associated pigments.

Acknowledgments

This research was supported by the Hundred-Talents Scheme of Chinese Academy of Sciences,

and the Hundred-Talents Scheme of Sun Yat-Sen University.

References

Aizawa, K., and Inakuma, T. (2007). Quantitation of carotenoids in commonly consumed vegetables

in Japan. Food Sci. Technol. Res., 13: 247-252.

Al-Duais, M., Hohbein, J., Werner, S., Bohm, V., and Jetschke, G. (2009). Contents of vitamin C,

carotenoids, tocopherols, and tocotrienols in the subtropical plant species Cyphostemma digitatum

as affected by processing. J. Agr. Food Chem., 57: 5420-5427.

Arnal, E., Miranda, M., Almansa, I. , Muriach, M., Barcia, J. M., Romero, F. J. , Diaz-Llopis, M., and

Bosch-Morell, F. (2009a). Lutein prevents cataract development and progression in diabetic rats.

Graef. Arch. Clin. Exp., 247: 115-120.

Arnal, E., Miranda, M. , Johnsen-Soriano, S., Alvarez-Nolting, R., Diaz-Llopis, M., Araiz, J., Cervera,

E., Bosch-Morell, F. , and Romero, F. J. (2009b). Beneficial effect of docosahexanoic acid and

lutein on retinal structural, metabolic, and functional abnormalities in diabetic rats. Curr. Eye Res.,

34: 928-938.

Arnal, E., Miranda, M., Barcia, J., Bosch-Morell, F., and Romero, F. J. (2010). Lutein and

docosahexaenoic acid prevent cortex lipid peroxidation in streptozotocin-induced diabetic rat

cerebral cortex. Neurosci., 166: 271-278.

Astner, S., Wu, A., Chen, J. , Philips, N., Rius-Diaz, F., Parrado, C., Mihm, M. C., Goukassian, D. A.,

Pathak, M. A., and Gonzalez, S. (2007). Dietary lutein/zeaxanthin partially reduces photoaging and

photocarcinogenesis in chronically UVB-irradiated Skh-1 hairless mice. Skin Pharmacol. Phys.,

20: 283-291.

Azevedo-Meleiro, C. H., and Rodriguez-Amaya, D. B. (2007). Qualitative and quantitative

differences in carotenoid composition among Cucurbita moschata, Cucurbita maxima, and

Cucurbita pepo. J. Agr. Food Chem., 55: 4027-4033.



88

Barker, F. M., Snodderly, D. M., Johnson, E. J., Schalch, W., Koepcke, W., Gerss, J., and M.

Neuringer, J. (2011). Nutritional manipulation of primate retinas, V: Effects of lutein, zeaxanthin,

and n-3 fatty acids on retinal sensitivity to blue-light-induced damage. Invest. Ophth. Vis. Sci., 52:

3934-3942.

Bartlett, H. E., and Eperjesi, F. (2007). Effect of lutein and antioxidant dietary supplementation on

contrast sensitivity in age-related macular disease: A randomized controlled trial. Eur. J. Clin.

Nutr., 61: 1121-1127.

Belefant-Miller, H., and Grace, S. C. (2010). Variations in bran carotenoid levels within and between

rice subgroups. Plant Food Hum. Nutr., 65: 358-363.

Beydoun, H. A., Shroff, M. R., Mohan, R., and Beydoun, M. A. (2011). Associations of serum

vitamin A and carotenoid levels with markers of prostate cancer detection among US men. Cancer

Cause Control, 22: 1483-1495.

Bhattacharyya, S., Roychowdhury, A., and Ghosh, S. (2008). Lutein content, fatty acid composition

and enzymatic modification of lutein from marigold (Tagetes patula L.) flower petals. J. Indian

Chem. Soc., 85: 942-944.

Bhattacharyya, S., Datta, S., Mallick, B., Dhar, P. , and Ghosh, S. (2010). Lutein content and in vitro

antioxidant activity of different cultivars of Indian marigold flower (Tagetes patula L.) extracts. J.

Agr. Food Chem., 58: 8259-8264.

Bone, R. A., Landrum, J. T., and Tarsis, S. L. (1985). Preliminary identification of the human macular

pigment. Vision Res., 25: 1531-1535.

Bonierbale, M., Gruneberg, W., Amoros, W., Burgos, G., Salas, E. , Porras, E., and zum Felde, T.

(2009). Total and individual carotenoid profiles in Solanum phureja cultivated potatoes: II.

Development and application of near-infrared reflectance spectroscopy (NIRS) calibrations for

germplasm characterization. J. Food Compos. Anal., 22: 509-516.

Briciu, R. D., Casoni, D., and Bischin, C. (2008). Thin layer chromatography separation of some

carotenoids, retinoids and tocopherols. Stud. U Babes-Bol. Che., 53: 67-75.

Bunea, A., Andjelkovic, M., Socaciu, C., Bobis, O., Neacsu, M., Verhe, R., and Van Camp, J. (2008).

Total and individual carotenoids and phenolic acids content in fresh, refrigerated and processed

spinach (Spinacia oleracea L.). Food Chem., 108: 649-656.

Cha, K. H., Koo, S. Y., and Lee, D. U. (2008). Antiproliferative effects of carotenoids extracted from

Chlorella ellipsoidea and Chlorella vulgaris on human colon cancer cells. J. Agr. Food Chem., 56:

10521-10526.



89

Chandrika, U. G., Basnayake, B. M. L. B., Athukorala, I., Colombagama, P. W. N. M., and

Goonetilleke, A. (2010). Carotenoid content and in vitro bioaccessibility of lutein in some leafy

vegetables popular in Sri Lanka. J. Nutr. Sci. Vitaminol., 56: 203-207.

Chandrika, U. G., Salim, N., Wijepala, G. D. D. J., Perera, K. S. U., and Goonetilleke, A. K. E. (2011).

Carotenoid and mineral content of different morphotypes of Centella asiatica L. (Gotukola). Int. J.

Food Sci. Nutr., 62: 552-557.

Chauveau-Duriot, B., Doreau, M., Noziere, P., and Graulet, B. (2010). Simultaneous quantification of

carotenoids, retinol, and tocopherols in forages, bovine plasma, and milk: Validation of a novel

UPLC method. Anal. Bioanal. Chem., 397: 777-790.

Chen, X. J., Wu, J. G., Zhou, S. J., Yang, Y. J., Ni, X. L., Yang, J. , Zhu, Z. J., and Shi, C. H. (2009).

Application of near-infrared reflectance spectroscopy to evaluate the lutein and β-carotene in

Chinese kale. J. Food Compos. Anal., 22: 148-153.

Cho, E. Y., Hankinson, S. E., Rosner, B., Willett, W. C., and Colditz, G. A. (2008). Prospective study

of lutein/zeaxanthin intake and risk of age-related macular degeneration. Am. J. Clin. Nutr., 87:

1837-1843.

Chong, E. W. T., Wong, T. Y., Kreis, A. J., Simpson, J. A., and Guymer, R. H. (2007). Dietary

antioxidants and primary prevention of age related macular degeneration: Systematic review and

meta-analysis. Brit. Med. J., 335: 755-759.

Chu, D. Y., Xie, Y. Q., Hong, S. Q., Yan, X. Y., Tong, H. W., and Liu, S. M. (2010). Major

carotenoids in tobacco laminas: Identification and quantification by HPLC with photodiode array

detection. Asian J. Chem., 22: 2635-2647.

Comstock, G. W., Alberg, A. J., Huang, H. Y., Wu, K., Burke, A. E., Hoffman, S. C., Norkus, E. P.,

Gross, M., Cutler, R. G., Morris, J. S., et al., (2008). The risk of developing lung cancer associated

with antioxidants in the blood: ascorbic acids, carotenoids. alpha-tocopherol, selenium, and total

peroxyl radical absorbing capacity (reprinted from cancer epidemiology, biomarkers & prevention,

vol 6, pg 907-916, 1997). Am. J. Epidemiol., 168: 831-840.

Crupi, P., Milella, R. A., and Antonacci, D. (2010). Simultaneous HPLC-DAD-MS (ESI+)

determination of structural and geometrical isomers of carotenoids in mature grapes. J. Mass

Spectrom., 45: 971-980.

Daly, T., Jiwan, M. A., O'Brien, N. M., and Aherne, S. A. (2010). Carotenoid content of commonly

consumed herbs and assessment of their bioaccessibility using an in vitro digestion model. Plant

Food Hum. Nutr., 65: 164-169.



90

Davey, M. W., Saeys, W., Hof, E., Ramon, H., Swennen, R. L., and Keulemans, J. (2009). Application

of visible and near-infrared reflectance spectroscopy (Vis/NIRS) to determine carotenoid contents

in banana (Musa sp.) fruit pulp. J. Agr. Food Chem., 57: 1742-1751.

de Faria, A. F., de Rosso, V. V., and Mercadante, A. Z. (2009). Carotenoid composition of jackfruit

(Artocarpus heterophyllus), determined by HPLC-PDA-MS/MS. Plant Food Hum. Nutr., 64: 108-

115.

Dias, M. G., Camoes, M. F. G. F. C., and Oliveira, L. (2009). Carotenoids in traditional Portuguese

fruits and vegetables. Food Chem., 113: 808-815.

Dugo, P., Herrero, M., Giuffrida, D., Kumm, T., Dugo, G., and Mondello, L. (2008). Application of

comprehensive two-dimensional liquid chromatography to elucidate the native carotenoid

composition in red orange essential oil. J. Agr. Food Chem., 56: 3478-3485.

Faria, A. F., Marques, M. C., and Mercadante, A. Z., (2011). Identification of bioactive compounds

from jambolao (Syzygium cumini) and antioxidant capacity evaluation in different pH conditions.

Food Chem., 126: 1571-1578.

Fu, H., Xie, B., Ma, S., Zhu, X., Fan, G., and Pan, S. (2011). Evaluation of antioxidant activities of

principal carotenoids available in water spinach (Ipomoea aquatica). J. Food Compos. Anal., 24:

288-297.

Gao, Y. X., Nagy, B., Liu, X., Simandi, B., and Wang, Q. (2009). Supercritical CO(2) extraction of

lutein esters from marigold (Tagetes erecta L.) enhanced by ultrasound. J. Supercrit. Fluid, 49:

345-350.

Gentili, A., and Caretti, F. (2011). Evaluation of a method based on liquid chromatography-diode array

detector-tandem mass spectrometry for a rapid and comprehensive characterization of the fat-

soluble vitamin and carotenoid profile of selected plant foods. J. Chromatogr. A, 1218: 684-697.

Ghosh, C., Baker, J. A., Moysich, K. B., Rivera, R., Brasure, J. R., and McCann, S. E. (2008). Dietary

intakes of selected nutrients and food groups and risk of cervical cancer. Nutr. Cancer, 60: 331-341.

Gomez-Prieto, M. S., Ruiz del Castillo, M. L., Flores, G., Santa-Maria, G., and Blanch, G. P. (2007).

Application of Chrastil's model to the extraction in SC-CO2 of β -carotene and lutein in Mentha

spicata L. J. Supercrit. Fluid, 43: 32-36.

Goodman, M. T., Shvetsov, Y. B., McDuffie, K., Wilkens, L. R., Zhu, X. M., Franke, A. A., Bertram,

C. C., Kessel, B., Bernice, M., Sunoo, C., et al. (2007). Hawaii cohort study of serum micronutrient

concentrations and clearance of incident oncogenic human papillomavirus infection of the cervix.

Cancer Res., 67: 5987-5996.

Granado-Lorencio, F., Herrero-Barbudo, C., Blanco-Navarro, I., and Perez-Sacristan, B. (2010).

Suitability of ultra-high performance liquid chromatography for the determination of fat-soluble



91

nutritional status (vitamins A, E, D, and individual carotenoids). Anal. Bioanal. Chem., 397: 1389-

1393.

Guil-Guerrero, J. L. and Rebolloso-Fuentes, M. M. (2008). Nutrient composition of Chlorella sp. and

Monodus subterraneus cultured in a bubble column bioreactor. Food Biotechnol., 22: 218-233.

Hayes, J. E., Allen, P., Brunton, N., O'Grady, M. N., and Kerry, J. P. (2011). Phenolic composition and

in vitro antioxidant capacity of four commercial phytochemical products: Olive leaf extract (Olea

europaea L.), lutein, sesamol and ellagic acid. Food Chem., 126: 948-955.

Hidalgo, A. and A. Brandolini (2008). Protein, ash, lutein and tocols distribution in einkorn (Triticum

monococcum L. subsp monococcum) seed fractions. Food Chem., 107: 444-448.

Hojnik, M., Skerget, M., and Knez, Z. (2008). Extraction of lutein from marigold flower petals -

Experimental kinetics and modelling. Lwt-Food Sci. and Technol., 41: 2008-2016.

Holasova, M., Dostalova, R., Fiedlerova, V., and Horacek, J. (2009). Variability of lutein content in

peas (Pisum sativum L.) in relation to the variety, season and chlorophyll content. Czech. J. Food

Sci., 27: S188-S191.

Horie, S., Okuda, C., Yamashita, T., Watanabe, K., Kuramochi, K., Hosokawa, M., Takeuchi, T.,

Kakuda, M., Miyashita, K., Sugawara, F., et al. (2010). Purified canola lutein selectively inhibits

specific isoforms of mammalian DNA polymerases and reduces inflammatory response. Lipids, 45:

713-721.

Hsu, Y. W., Tsai, C. F., Chen, W. K., Ho, Y. C., and Lu, F. J. (2011). Determination of lutein and

zeaxanthin and antioxidant capacity of supercritical carbon dioxide extract from daylily

(Hemerocallis disticha). Food Chem., 129: 1813-1818.

Hu, B. J., Hu, Y. N., Lin, S., Ma, W. J., and Li, X. R. (2011). Application of lutein and zeaxanthin

nonproliferative diabetic retinopathy. Int. J. Ophthalmol-Chi., 4: 303-306.

Hu, Y. Z., and Xu, Z. R. (2008). Effects of lutein on the growth and migration of bovine lens epithelial

cells in vitro. J Huazhong U Sci-Med, 28: 360-363.

Huang, X. Y., Liu, Y. W., Di, D. L., Liu, J. X., and Li, C. (2010). An improved LC-DAD method for

simultaneous determination of lutein, β-carotene and lycopene in tomato and its products.

Chromatographia, 71: 331-334.

Ishida, B. K., and Chapman, M. H. (2009). Carotenoid extraction from plants using a novel,

environmentally friendly solvent. J. Agr. Food Chem., 57: 1051-1059.

Johnson, E. J., Chung, H. Y., Caldarella, S. M., and Snodderly, D. M. (2008). The influence of

supplemental lutein and docosahexaenoic acid on serum, lipoproteins, and macular pigmentation.

Am. J. Clin. Nutr., 87: 1521-1529.



92

Kamffer, Z., Bindon, K. A., and Oberholster, A. (2010). Optimization of a method for the extraction

and quantification of carotenoids and chlorophylls during ripening in grape berries (Vitis vinifera

cv. Merlot). J. Agr. Food Chem., 58: 6578-6586.

Karlina, M. V., Pozharitskaya, O. N., Shikov, A. N., Kosman, V. M., Makarova, M. N., and Makarov,

V. G. (2008). LC method for quantification of lutein in rat plasma: Validation, and application to a

pharmacokinetic study. Chromatographia, 68: 949-954.

Kim, Y. N., Giraud, D. W., and Driskell, J. A. (2007). Tocopherol and carotenoid contents of selected

Korean fruits and vegetables. J. Food Compos. Anal., 20: 458-465.

Kim, J. H., Na, H. J., Kim, C. K., Kim, J. Y., Ha, K. S., Lee, H., Chung, H. T., Kwon, H. J., Kwon, Y.

G., and Kim, Y. M. (2008). The non-provitarnin A carotenoid, lutein, inhibits NF-kappa B-

dependent gene expression through redox-based regulation of the phosphatidylinositol 3-

kinase/PTEN/Akt and NF-kappa B-inducing kinase pathways: Role of H2O2 in NF-kappa B

activation. Free Radic. Biol. Med., 45: 885-896.

Kim, J. E., Leite, J. O., de Ogburn, R., Smyth, J. A., Clark, R. M., and Fernandez, M. L. (2011a). A

lutein-enriched diet prevents cholesterol accumulation and decreases oxidized LDL and

inflammatory cytokines in the aorta of guinea pigs. J. Nutr., 141: 1458-1463.

Kim, Y., Seo, J. H., and Kim, H. (2011b). Beta-carotene and lutein inhibit hydrogen peroxide-induced

activation of NF-kappa B and IL-8 expression in gastric epithelial AGS cells. J. Nutr. Sci.

Vitaminol., 57: 216-223.

Kimura, M., Kobori, C. N., Rodriguez-Amaya, D. B., and Nestel, P. (2007). Screening and HPLC

methods for carotenoids in sweetpotato, cassava and maize for plant breeding trials. Food Chem.,

100: 1734-1746.

Kitada, K., Machmudah, S., Sasaki, M., Goto, M., Nakashima, Y., Kumamoto, S., and Hasegawa, T.

(2009). Supercritical CO2 extraction of pigment components with pharmaceutical importance from

Chlorella vulgaris. J. Chem. Technol. Biot., 84: 657-661.

Kobori, C. N., and Arnaya, D. B. R. (2008). Uncultivated Brazilian green leaves are richer sources of

carotenoids than are commercially produced leafy vegetables. Food Nutr. Bull., 29: 320-328.

Koh, W. P., Yuan, J. M., Wang, R., Lee, Y. P., Lee, B. L., Yu, M. C., and Ong, C. N. (2011). Plasma

carotenoids and risk of acute myocardial infarction in the Singapore Chinese health study. Nutr.

Metab. Cardiovas., 21: 685-690.

Krishnaswamy, R., Devaraj, S. N., and Padma, V. V. (2010). Lutein protects HT-29 cells against

Deoxynivalenol-induced oxidative stress and apoptosis: Prevention of NF-kappa B nuclear

localization and down regulation of NF-kappa B and Cyclo-Oxygenase 2 expression. Free Radic.

Biol. Med., 49: 50-60.



93

Kumar, R., Yu, W. L., Jiang, C. L., Shi, C. L., and Zhao, Y. P. (2010). Improvement of the isolation

and purification of lutein from marigold flower (Tagetes Erecta L.) and its antioxidant activity. J.

Food Process. Eng., 33: 1065-1078.

Kupper, H., Seibert, S., and Parameswaran, A. (2007). Fast, sensitive, and inexpensive alternative to

analytical pigment HPLC: Quantification of chlorophylls and carotenoids in crude extracts by

fitting with gauss peak spectra. Anal. Chem., 79: 7611-7627.

Li, L., Qin, J., Yin, S., and Tang, G. W. (2007). Effects of mobile phase ratios on the separation of

plant carotenoids by HPLC with a C30 column. Chromatographia, 65: 91-94.

Li, S. Y., Fu, Z. J., Ma, H., Jang, W. C., So, K. F., Wong, D., and Lo, A. C. Y. (2009). Effect of lutein

on retinal neurons and oxidative stress in a model of acute retinal ischemia/reperfusion. Invest.

Ophth. Vis. Sci., 50: 836-843.

Li, B. X., Ahmed, F., and Bernstein, P. S. (2010a). Studies on the singlet oxygen scavenging

mechanism of human macular pigment. Arch. Biochem. Biophys., 504: 56-60.

Li, L., Chen, C. Y. O., Aldini, G., Johnson, E. J., Rasmussen, H., Yoshida, Y., Niki, E., Blumberg, J.

B., Russell, R. M., and Yeum, K. J. (2010b). Supplementation with lutein or lutein plus green tea

extracts does not change oxidative stress in adequately nourished older adults. J. Nutr. Biochem.,

21: 544-549.

Li, X. X., and Han, L. J. (2009). Effect of enzymatic treatment of corn gluten meal on lutein,

zeaxanthin and beta-crytoxanthin extraction. J. Food Process Eng., 32: 206-220.

Li, S. Y., and Lo, A. C. Y. (2010). Lutein protects RGC-5 cells against hypoxia and oxidative stress.

Int. J. Mol. Sci., 11: 2109-2117.

Liu, Y. T., Perera, C. O., and Suresh, V. (2007). Comparison of three chosen vegetables with others

from South East Asia for their lutein and zeaxanthin content. Food Chem., 101: 1533-1539.

Lozano, S. E., Suarez, D. U., Diaz, B. H. C., Ocampo, M. L. A., Martinez, A. A. D., Jimenez, F. G.,

and Aparicio, A. J. (2011). Study of development, quality of plant and accumulation of pigments in

ligules of Tagetes erecta L. exposed to two light intensities. B Latinoam. Caribe. Pl., 10: 476-488.

Machmudah, S., Kawahito, Y., Sasaki, M., and Goto, M. (2008). Process optimization and extraction

rate analysis of carotenoids extraction from rosehip fruit using supercritical CO2. J. Supercrit.

Fluid., 44: 308-314.

Maci, S. (2007). Nutritional support against skin ageing: New scientific evidence for the role of lutein.

Agro. Food Ind. Hi Tec., 18: 42-44.

Macias-Sanchez, M. D., Fernandez-Sevilla, J. M., Fernandez, F. G. A., Garcia, M. C. C., and Grima,

E. M. (2010). Supercritical fluid extraction of carotenoids from Scenedesmus almeriensis. Food

Chem., 123: 928-935.



94

Mamatha, B. S., Sangeetha, R. K., and Baskaran, V. (2011). Provitamin-A and xanthophyll

carotenoids in vegetables and food grains of nutritional and medicinal importance. Int. J. Food Sci.

Tech., 46: 315-323.

Mignone, L. I., Giovannucci, E., Newcomb, P. A., Titus-Ernstoff, L., Trentham-Dietz, A., Hampton, J.

M., Willett, W. C., and Egan, K. M. (2009). Dietary carotenoids and the risk of invasive breast

cancer. Int. J. Cancer, 124: 2929-2937.

Miguel, F., Martin, A., Mattea, F., and Cocero, M. J. (2008). Precipitation of lutein and co-

precipitation of lutein and poly-lactic acid with the supercritical anti-solvent process. Chem. Eng.

Proc., 47: 1594-1602.

Moeller, S. M., Voland, R., Tinker, L., Blodi, B. A., Klein, M. L., Gehrs, K. M., Johnson, E. J.,

Snodderly, M., Wallace, R. B., Chappell, R. J., et al. (2008). Associations between age-related

nuclear cataract and lutein and zeaxanthin in the diet and serum in the carotenoids in the age-related

eye disease study (CAREDS), an ancillary study of the women's health initiative. Arch Ophthalmol-

Chic., 126: 354-364.

Moreno, F. S., Toledo, L. P., de Conti, A., Heidor, R., Jordo, A., Vannucchi, H., Cardozo, M. T., and

Ong, T. P. (2007). Lutein presents suppressing but not blocking chernopreventive activity during

diethylnitrosamine-induced hepatocarcinogenesis and this involves inhibition of DNA damage.

Chem-Biol. Interact., 168: 221-228.

Muriach, M., Bosch-Morell, F., Arnal, E., Alexander, G., Blomhoff, R., and Romero, F. J. (2008).

Lutein prevents the effect of high glucose levels on immune system cells in vivo and in vitro. J.

Physiol. Biochem., 64: 149-157.

Murillo, E., Melendez-Martinez, A. J., and Portugal, F. (2010). Screening of vegetables and fruits from

Panama for rich sources of lutein and zeaxanthin. Food Chem., 122: 167-172.

Palombo, P., Fabrizi, G., Ruocco, V., Ruocco, E., Fluhr, J., Roberts, R., and Morganti, P. (2007).

Beneficial long-term effects of combined oral/topical antioxidant treatment with the carotenoids

lutein and zeaxanthin on human skin: A double-blind, placebo-controlled study. Skin Pharmacol.

Phys., 20: 199-210.

Pang, R., Tao, J. Y., Zhang, S. L., Zhao, L., Yue, X., Wang, Y. F., Ye, P. A., Dong, J. H., Zhu, Y., and

Wu, J. G. (2010). In vitro antiviral activity of lutein against hepatitis B virus. Phytother Res., 24:

1627-1630.

Pelucchi, C., Dal Maso, L., Montella, M., Parpinel, M., Negri, E., Talamini, R., Giudice, A.,

Franceschi, S., and La Vecchia, C. (2008). Dietary intake of carotenoids and retinol and

endometrial cancer risk in an Italian case-control study. Cancer Cause Control, 19: 1209-1215.



95

Perry, A., Rasmussen, H., and Johnson, E. J. (2009). Xanthophyll (lutein, zeaxanthin) content in fruits,

vegetables and corn and egg products. J. Food Compos. Anal., 22: 9-15.

Pham, V. H., and Hatcher, D. W. (2011). Ultra-performance liquid chromatography (UPLC)

quantification of carotenoids in durum wheat influence of genotype and environment in relation to

the colour of yellow alkaline noodles (YAN). Food Chem., 125: 1510-1516.

Pintea, A., Rugina, D. O., Pop, R., Bunea, A., and Socaciu, C. (2011). Xanthophylls protect against

induced oxidation in cultured human retinal pigment epithelial cells. J. Food Compos. Anal., 24:

830-836.

Pinto, E., Carvalho, A. P., Cardozo, K. H. M., Malcata, F. X., dos Anjos, F. M., and Colepicolo, P.

(2011). Effects of heavy metals and light levels on the biosynthesis of carotenoids and fatty acids in

the macroalgae Gracilaria tenuistipitata (var. liui Zhang & Xia). Rev. Bras. Farmacogn., 21: 349-

354.

Provesi, J. G., Dias, C. O., and Amante, E. R. (2011). Changes in carotenoids during processing and

storage of pumpkin puree. Food Chem., 128: 195-202.

Rafi, M. M., and Shafaie, Y. (2007). Dietary lutein modulates inducible nitric oxide synthase (iNOS)

gene and protein expression in mouse macrophage cells (RAW 264.7). Mol. Nutr. Food Res., 51:

333-340.

Ren, D., and Zhang, S. H. (2008). Separation and identification of the yellow carotenoids in

Potamogeton crispus L. Food Chem., 106: 410-414.

Reynoso-Camacho, R., Gonzalez-Jasso, E., Ferriz-Martinez, R., Villalon-Corona, B., Loarca-Pina, G.

F., Salgado, L. M., and Ramos-Gomez, M. (2011). Dietary supplementation of lutein reduces colon

carcinogenesis in DMH-treated rats by modulating K-ras, PKB, and β-catenin proteins. Nutr.

Cancer, 63: 39-45.

Rivera, S., Vilaro, F., and Canela, R. (2011). Determination of carotenoids by liquid

chromatography/mass spectrometry: Effect of several dopants. Anal. Bioanal. Chem., 400: 1339-

1346.

SanGiovanni, J. P., Chew, E. Y., Clemons, T. E., Ferris, F. L., Gensler, G., Linblad, A. S., Milton, R.

C., Seddon, J. M., and Sperduto, R. D. (2007). The relationship of dietary carotenoid and vitamin A,

E, and C intake with age-related macular degeneration in a case-control study - AREDS Report no.

22. Arch. Ophthalmol-Chic., 125: 1225-1232.

Santocono, M., Zurria, M., Berrettini, M., Fedeli, D., and Falcioni, G. (2007). Lutein, zeaxanthin and

astaxanthin protect against DNA damage in SK-N-SH human neuroblastoma cells induced by

reactive nitrogen species. J. Photoch. Photobio. B, 88: 1-10.



96

Sasaki, M., Ozawa, Y., Kurihara, T., Noda, K., Imamura, Y., Kobayashi, S., Ishida, S., and Tsubota, K.

(2009). Neuroprotective effect of an antioxidant, lutein, during retinal inflammation. Invest. Ophth.

Vis. Sci., 50: 1433-1439.

Sasaki, M., Ozawa, Y., Kurihara, T., Kubota, S., Yuki, K., Noda, K., Kobayashi, S., Ishida, S., and

Tsubota, K. (2010). Neurodegenerative influence of oxidative stress in the retina of a murine model

of diabetes. Diabetologia, 53: 971-979.

Satia, J. A., Littman, A., Slatore, C. G., Galanko, J. A., and White, E. (2009). Long-term use of β-

carotene, retinol, lycopene, and lutein supplements and lung cancer risk: Results from the VITamins

And Lifestyle (VITAL) study. Am. J. Epidemiol., 169: 815-828.

Sato, Y., Kobayashi, M., Itagaki, S., Hirano, T., Noda, T., Mizuno, S., Sugawara, M., and Iseki, K.

(2011). Protective effect of lutein after ischemia-reperfusion in the small intestine. Food Chem.,

127: 893-898.

Scarmo, S., Cartmel, B., Lin, H. Q., Leffell, D. J., Welch, E., Bhosale, P., Bernstein, P. S., and Mayne,

S. T. (2010). Significant correlations of dermal total carotenoids and dermal lycopene with their

respective plasma levels in healthy adults. Arch. Biochem. Biophys., 504: 34-39.

Serpeloni, J. M., Grotto, D., Mercadante, A. Z., Bianchi, M. D. P., and Antunes, L. M. G. (2010).

Lutein improves antioxidant defense in vivo and protects against DNA damage and chromosome

instability induced by cisplatin. Arch. Toxicol., 84: 811-822.

Shi, J., Yi, C., Ye, X. Q., Xue, S., Jiang, Y. M., Ma, Y., and Liu, D. H. (2010). Effects of supercritical

CO2 fluid parameters on chemical composition and yield of carotenoids extracted from pumpkin.

LWT-Food Sci.Technol., 43: 39-44.

Shvetsov, Y. B., Hernandez, B. Y., Wilkens, L. R., Thompson, P. J., Franke, A. A., Zhu, X. M., and

Goodman, M. T. (2010). Plasma micronutrients and the acquisition and clearance of anal human

papillomavirus infection: The Hawaii HPV cohort study. Cancer Res., 70: 9787-9797.

Sindhu, E. R., Firdous, A. P., Preethi, K. C., and Kuttan, R. (2010). Carotenoid lutein protects rats

from paracetamol-, carbon tetrachloride- and ethanol-induced hepatic damage. J. Pharm.

Pharmacol., 62: 1054-1060.

Skerget, M., Bezjak, M., Makovsek, K., and Knez, Z. (2010). Extraction of lutein diesters from tagetes

erecta using supercritical CO2 and liquid propane. Acta Chim. Slov., 57: 60-65.

Sun, Z., Liu, J., Zeng, X. H., Huangfu, J. Q., Jiang, Y., Wang, M. F., and Chen, F. (2011). Protective

actions of microalgae against endogenous and exogenous advanced glycation endproducts (AGEs)

in human retinal pigment epithelial cells. Food Funct., 2: 251-258.



97

Tlili, N., Nasri, N., Saadaoui, E., Khaldi, A., and Triki, S. (2009). Carotenoid and tocopherol

composition of leaves, buds, and flowers of Capparis spinosa grown wild in Tunisia. J. Agr. Food

Chem., 57: 5381-5385.

Toiu, A., Muntean, E., Oniga, I., and Tamas, M. (2009). Pharmacognostic research on Viola declinata

Waldst. Et Kit. (Violaceae). Farmacia, 57: 218-222.

Tsai, Y. C., Wu, W. B., and Chen, B. H. (2010). Preparation of carotenoids and chlorophylls from

Gynostemma pentaphyllum (Thunb.) Makino and their antiproliferation effect on hepatoma cell. J.

Med. Food, 13: 1431-1442.

Tsuboi, M., Etoh, H., Yomoda, Y., Kato, K., Kato, H., Kulkarni, A., Terada, Y., Maoka, T., Mori, H.,

and Inakuma, T. (2010). Nitration reaction of lutein with peroxynitrite. Tetrahedron. Lett., 51: 676-

678.

Wang, Y. C., Chuang, Y. C., and Hsu, H. W. (2008). The flavonoid, carotenoid and pectin content in

peels of citrus cultivated in Taiwan. Food Chem., 106: 277-284.

Wang, X. C., Chen, L., Ma, C. L., Yao, M. Z., and Yang, Y. J. (2010). Genotypic variation of β-

carotene and lutein contents in tea germplasms, Camellia sinensis (L.) O. Kuntze. J. Food Compos.

Anal., 23: 9-14.

Weigert, G., Kaya, S., Pemp, B., Sacu, S., Lasta, M., Werkmeister, R. M., Dragostinoff, N., Simader,

C., Garhofer, G., Schmidt-Erfurth, U., et al. (2011). Effects of lutein supplementation on macular

pigment optical density and visual acuity in patients with age-related macular degeneration. Invest.

Ophth. Vis. Sci., 52: 8174-8178.

Yahia, E. M., Gutierrez-Orozco, F., and Arvizu-de Leon, C. (2011). Phytochemical and antioxidant

characterization of mamey (Pouteria sapota Jacq. HE Moore & Stearn) fruit. Food Res. Int., 44:

2175-2181.

Yang, J., Zhu, Z. J., Wang, Z. Z., and Zhu, B. A. (2010). Effects of storage temperature on the contents

of carotenoids and glucosinolates in Pakchoi (Brassica Rapa L. Ssp. Chinensis Var. Communis). J.

Food Biochem., 34: 1186-1204.

Yue, X., Xu, Z., Prinyawiwatkul, W., and King, J. M. (2006). Improving extraction of lutein from egg

yolk using an ultrasound-assisted solvent method. J. Food Sci., 71: C239-C241.

Zanatta, C. F., and Mercadante, A. Z. (2007). Carotenoid composition from the Brazilian tropical fruit

camu-camu (Myrciaria dubia). Food Chem., 101: 1526-1532.

Zhang, J. J., Dhakal, I., Stone, A., Ning, B. T., Greene, G., Lang, N. P., and Kadlubar, F. F. (2007a).

Plasma carotenoids and prostate cancer: A population-based case-control study in Arkansas. Nutr.

Cancer, 59: 46-53.



98

Zhang, M., D'Arcy, C., Holman, J., and Binns, C. W. (2007b). Intake of specific carotenoids and the

risk of epithelial ovarian cancer. Brit. J. Nutr., 98: 187-193.

Zhang, M., Hettiarachchy, N. S., Horax, R., Chen, P. Y., and Over, K. F. (2009). Effect of maturity

stages and drying methods on the retention of selected nutrients and phytochemicals in bitter melon

(Momordica charantia) leaf. J. Food Sci., 74: C441-C448.

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