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Lutein as a functional food ingredient: Stability and bioavailability

March 2020
Journal of Functional Foods 66

DOI:10.1016/j.jff.2019.103771

Authors:

Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco

Lutein is a carotenoid found in numerous organisms ranging from bacteria to algae, yeasts, and plants. The biological importance of this pigment emerged from various studies demonstrating its antioxidant potential, playing a significant role in the prevention of age-related macular degeneration and other diseases such as cancer. Lutein is also essential in the development of infants brain and must be consumed in sufficient amounts to obtain its health benefits. Lutein biologically effective dose is difficult to achieve due to its low lutein bioaccessibility and bioavailability in food sources. These factors are differentially affected by the properties of the food matrix, the processing, and the presence of other dietary components. The objective of this literature review was to explore lutein stability and bioavailability by modifications and variations associated with food technological procedures.

Isoprene molecules (A) can bond together (B) to form tetraterpenes, structures made up of 8 isoprene units. Carotenoids are tetraterpenes divided into the carotenes (C) and xanthophylls (D).

…

Structure of all-trans lutein (A). It can be esterified with fatty acids, including palmitic acid (B), and its oxidation products are usually apocarotenoids (C).

…

In vivo studies evaluating the bioavailability of lutein.

…

Condition and matrix effect on lutein stability.

…

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Contents lists available at ScienceDirect

Journal of Functional Foods

journal homepage: www.elsevier.com/locate/jff

Lutein as a functional food ingredient: Stability and bioavailability

Mario Ochoa Becerra

, Luis Mojica Contreras

, Ming Hsieh Lo

, Juan Mateos Díaz

Gustavo Castillo Herrera

a,⁎

Tecnología Alimentaria, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A.C. CIATEJ, Unidad Zapopan, Camino Arenero 1227, El Bajío

45019, Zapopan, Jalisco, Mexico

Biotecnología Industrial, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A.C. CIATEJ, Unidad Zapopan, Camino Arenero 1227, El

Bajío 45019, Zapopan, Jalisco, Mexico

ARTICLE INFO

Keywords:

Bioaccessibility

Bioavailability

Lutein

Stability

Food matrix

Dietary source

ABSTRACT

Lutein is a carotenoid found in numerous organisms ranging from bacteria to algae, yeasts, and plants. The

biological importance of this pigment emerged from various studies demonstrating its antioxidant potential,

playing a signiﬁcant role in the prevention of age-related macular degeneration and other diseases such as

cancer. Lutein is also essential in the development of infants brain and must be consumed in suﬃcient amounts

to obtain its health beneﬁts. Lutein biologically eﬀective dose is diﬃcult to achieve due to its low lutein

bioaccessibility and bioavailability in food sources. These factors are diﬀerentially aﬀected by the properties of

the food matrix, the processing, and the presence of other dietary components. The objective of this literature

review was to explore lutein stability and bioavailability by modiﬁcations and variations associated with food

technological procedures.

1. Introduction

Carotenoids are among the most abundant types of naturally oc-

curring fat-soluble pigments. Generally found in yellow-orange fruits,

marigold ﬂowers and dark green leafy vegetables, playing an essential

role on photosynthesis and photoprotection (Maiani et al., 2009; Walsh,

Bartlett, & Eperjesi, 2015). They are tetraterpenes made up of 8 iso-

prene units (Fig. 1A) containing extended conjugated double bond

systems (Fig. 1B), responsible for conferring color as well as strong

antioxidant activity. (Hajare, Ray, Shreya, Avadhani, & Selvaraj, 2013).

Carotenoids are divided into two groups: the carotenes (Fig. 1C), which

can be unsaturated hydrocarbons having the formula C

and the

xanthophylls (Fig. 1D); oxygenated derivatives of carotenes (Gong &

Bassi, 2016). The global market for carotenoids is continuously

growing; it has been valued at approximately US $1400 million in 2017

and should reach $2000 million by 2022, at an annual growth rate of

5.7% for the period of 2017–2022. One of the dominating carotenoids

in this market is lutein, with a 23% market share; lutein has the highest

growth potential of carotenoids (BCC Research, 2018; Del Campo,

García-González, & Guerrero, 2007). Carotenoids are essential pigments

for all photosynthetic organisms, they are found in plant tissues such as

leaves, roots, ﬂowers, and fruits. The current commercial source of

lutein is the ﬂower of the genus Tagetes; its petals are rich in lutein and

lutein fatty acid esters, which represent over 90% of the pigments

identiﬁed in this plant (Sandmann, 2015). Nevertheless, microalgae

have gained attention in this ﬁeld due to their high lutein content and

biomass productivity, making them a potential alternative source of this

carotenoid (Fernández-Sevilla, Acién Fernández, & Molina Grima,

2010).

Lutein form part of the xanthophyll family of carotenoids; it is

usually found in ﬂowers, grains, fruits and vegetables, such as spinach

and kale (Yang et al., 2018). The lutein market is segmented into

pharmaceutical, dietary supplement, food, and animal and ﬁsh feed

industries. Luitein principal application is to brighten the colors of

poultry feathers and deepen the yellow of egg yolk (Lin, Lee, & Chang,

2015). However, lutein use in the food industry is limited due to its

instability and the chemical changes caused during food processing

(Qv, Zeng, & Jiang, 2011). Processes which may aﬀect the integrity of

lutein include high temperatures, the presence of oxygen, light and

extreme pH (Gouveia & Empis, 2003).

During processing, foods require to be exposed to the high tem-

peratures of cooking by boiling, sauteing or steaming. Also, extreme pH

below 4.0 or above 8.0 can induce de-esteriﬁcation and cis/trans iso-

merization of the molecule. The presence of light may cause the for-

mation of colorless compounds of low molecular weight (Boon,

McClements, Weiss, & Decker, 2010; Cheng, Ferruzzi, & Jones, 2019;

https://doi.org/10.1016/j.jﬀ.2019.103771

Received 23 November 2018; Received in revised form 3 December 2019; Accepted 28 December 2019

⁎

Corresponding author.

E-mail address: gcastillo@ciatej.mx (G. Castillo Herrera).

Journal of Functional Foods 66 (2020) 103771

Available online 21 January 2020

(http://creativecommons.org/licenses/BY/4.0/).

Weigel, Weiss, Decker, & McClements, 2018). For this reason, it is es-

sential that formulations are engineered to ensure lutein stability

during ﬁnal product storage (Boon et al., 2010). Techniques such as

freezing, the inclusion of antioxidants and exclusion of oxygen in va-

cuum sealed and airtight containers decrease the loss of the carotenoid

during processing and storage of foods (Gouveia & Empis, 2003).

Lutein is considered an eﬀective functional compound beneﬁting

human health with the potential to ameliorate cardiovascular diseases

(Dwyer et al., 2001), various types of cancer (Demmig-Adams & Adams,

2002; Heber & Lu, 2002) and age-related macular degeneration (AMD)

(Landrum & Bone, 2001) due to its antioxidant potential. For this

reason, it is essential to consider two key aspects; maintaining lutein

integrity during food processing, and increasing accessibility in the food

matrix. Bioaccessibility determines the total amount that is available

for absorption during digestion (O’Neill et al., 2001), while the portion

of the compound entering the systemic circulation to participate in

physiological functions is known as bioavailability (Amorim-Carrilho,

Cepeda, Fente, & Regal, 2014; Kopec & Failla, 2018).Several factors

inﬂuence the level of both bioaccessibility and bioavailability including

food matrix composition, processing level, and interaction with other

dietary compounds (Yonekura & Nagao, 2007).

Moreover, a critical factor that signiﬁcantly inﬂuences the bioa-

vailability of lutein is the absorption rate. If a good absorption is carried

out in the body, a lower dose will be required and, therefore, the for-

mulation will be more cost-eﬀective. Taking this into account, it is es-

sential to consider the matrix in which the molecule will be in-

corporated since it can promote or decrease its bioaccessibility in the

gastrointestinal tract (GIT) (O’Neill et al., 2001). Furthermore, in order

to observe a decrease in AMD, a dose of ~6 mg/day of lutein needs to

be consumed; this is diﬃcult to achieve in a regular human diet due to

the low concentration and bioavailability of lutein in fruits and vege-

tables (Khachik, Bernstein, & Garland, 1997; Rodríguez-Bernaldo de

Quirós & Costa, 2006).

Consequently, emulsion-based delivery systems for lipophilic

bioactive agents have been developed, which include oil-in-water

emulsions, nano-emulsions, microencapsulation, and liposomes (Weigel

et al., 2018). These strategies can be designed to increase the bioa-

vailability, and inhibit the chemical degradation of carotenoids, main-

taining the physical properties of the system, isolating other food

components and improving absorption in the GIT (Qv et al., 2011).

Taking this into consideration, this work aimed to review common

methods applied in food processing and their eﬀects on the stability and

bioavailability of lutein integrated into a food matrix. Furthermore,

preservation techniques currently employed to maintain the integrity of

the product were also reviewed.

2. Carotenoids

Carotenoids are an extensive group of lipophilic yellow-orange

pigments derivatives of tetraterpenes, meaning they contain 40 carbon

atoms. They are considered the most widespread pigments found in

nature (Amorim-Carrilho et al., 2014; Sandmann, 2015) since they are

essential in photosynthetic organisms. Carotenoids are components of

light-harvesting pigment-protein complexes and play a role in cell-

protective mechanisms. Almost all carotenoids show scavenging prop-

erties against excessive numbers of free radicals that may be produced

throughout a cell life cycle. This antioxidant capacity can potentially

protect humans from the compromised immune response, premature

aging, macular degeneration, cardiovascular diseases, and arthritis

(Maiani et al., 2009).

Carotenoids can be classiﬁed into two groups, xanthophylls (e.g.,

lutein and zeaxanthin) and carotenes (e.g., β-carotene, lycopene).

Xanthophylls contain oxygen atoms in their chemical structure, while

carotenes are purely hydrocarbons. The oxygen present in xanthophylls

is either as hydroxyl groups and/or as epoxides, for this reason, their

higher polarity allows them to be more easily separated from carotenes

in chromatographic procedures (Amorim-Carrilho et al., 2014).

3. Lutein

Pure lutein typically appears as a yellow-orange crystalline, lipo-

philic, solid with the chemical name β,ε-carotene-3,3′-diol (C

)

As all carotenoids, lutein contains a backbone of conjugated carbon-

carbon double bonds, which allow free electron movement and causes

the absorbance of light in the blue region of the visible spectrum giving

it a strong yellow-orange color (Mora-Gutierrez et al., 2018; Yi, Fan,

Yokoyama, Zhang, & Zhao, 2016). Lutein generally coexists in nature

with its stereoisomer zeaxanthin and the double bonds of the isoprene

backbone can exist in the all-trans (Fig. 2A) or cis conﬁgurations, also

called E/Z conformations. In nature, the most common geometric

isomer of lutein is the all-trans (all-E) isomer being thermodynamically

more stable than the cis (Yang et al., 2018).

Lutein can be found as esteriﬁed or non-esteriﬁed with fatty acids,

most commonly with palmitic acid (Fig. 2B) (Abdel-Aal & Rabalski,

2015). However, esteriﬁed carotenoids cannot be adequately absorbed by

the GIT, but are less susceptible to degradation compared to their free

forms (Breithaupt, Bamedi, & Wirt, 2002; Martínez-Delgado, Khandual, &

Villanueva-Rodríguez, 2017). In marigold ﬂowers, lutein is mainly found

in its esteriﬁed form, and after extraction with solvents, it needs to be

further puriﬁed to its free lutein form. This is performed via reaction with

a strong base (e.g., KOH) in alcohols of low molecular weight such as

ethanol (Clowutimon, Shotipruk, Boonnoun, & Ponpesh, 2018).

Due to the intense yellow color lutein presents, it is widely used as a

natural food colorant. In recent years, these compounds have gained

greater attention because of their potential health beneﬁts. The low

stability of natural color additives is a signiﬁcant deterrent which has

slowed down their integration into the food industry. Increasing lutein

stability through various methods is being advocated to minimize this

problem (Rodriguez-Amaya, 2016).

4. Biological activity

The primary health beneﬁt of carotenoids is their strong antioxidant

potential (Fiedor & Burda, 2014). Additionally, speciﬁc carotenoids

may have additional beneﬁts such as β-carotene, having the ability to

act as a pro-vitamin A (Gurmu, Hussein, & Laing, 2014). Lutein and

zeaxanthin can protect the eye from UV radiation and are essential to

brain developing (Barker et al., 2011; Tanaka, Shnimizu, & Moriwaki,

2012). Other carotenoids may have the ability to help prevent heart

disease by blocking the formation of low-density lipoprotein

(Eggersdorfer & Wyss, 2018; Iwamoto et al., 2000; Yoshida et al.,

2010).

Fig. 1. Isoprene molecules (A) can bond together (B) to form tetraterpenes,

structures made up of 8 isoprene units. Carotenoids are tetraterpenes divided

into the carotenes (C) and xanthophylls (D).

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

4.1. Eye health

The macula is a specialized structure in the retina of humans and

primates; its center is the region of highest visual acuity, referred to as

the foveola. In the macula, lutein and zeaxanthin are selectively accu-

mulated and act as potent biological antioxidants. They serve as eﬃ-

cient blue-light ﬁlters, quenching reactive oxygen species (ROS) formed

during photoexcitation (Nwachukwu, Udenigwe, & Aluko, 2016;

Panova et al., 2017). Degenerative diseases can aﬀect the macula,

especially in individuals with more than 65 years old. This fact has

increased the need of including lutein in the diet. Consumption of lu-

tein-rich foods can lower the risk of developing AMD and cataracts

(Abdel-Aal, Akhtar, Zaheer, & Ali, 2013; Barker et al., 2011; Moeller

et al., 2008; Nwachukwu et al., 2016).

It is important to note that before lutein is accumulated in the retina

to produce beneﬁcial eﬀects, it must ﬁrst be released from the food

matrix, absorbed by the enterocytes in the intestine and then trans-

ported to the target tissue. Because of lutein lipophilic nature, it must

be incorporated into mixed micelles before absorption. It is of great

interest to increase the amount of lutein that is transferred from the

food matrix to micelles during the development of lutein bioactive

foods. Promoting a positive impact on the protection of eye health not

only in adults, but also in children and during pregnancy (Panova et al.,

2017; Xavier, Carvajal-Lérida, Garrido-Fernández, & Pérez-Gálvez,

2018).

4.2. Pro-Vitamin A function

In developing countries, Vitamin A (retinol) deﬁciency is a major

health concern aﬀecting primarily preschool children and pregnant

women; this can mainly cause blindness, poor growth, and death

(Gurmu et al., 2014; World Health Organization, 2002). Including

major sources of carotenoids in the regular diet can help to reduce this

problem because they can be transformed into vitamin A. An approach

to combat Vitamin A deﬁciency, is to fortify commonly used food items

such as sugars, fats, and cereal products by adding carotenoids. How-

ever, other factors aﬀects carotenoid conversion to retinol, such as

genetics and polymorphisms (Eggersdorfer & Wyss, 2018; Gurmu et al.,

2014; Weber & Grune, 2012).

4.3. Cardiovascular disease

Factors such as oxidative stress, inﬂammation, and dyslipidemia are

implicated in the development of cardiovascular disease, and there is

evidence that lutein may have beneﬁcial eﬀects in this regard (Böhm,

2012; Müller, Caris-Veyrat, Lowe, & Böhm, 2016). Melo van Lent et al.

(2016) suggested that higher dietary intake and blood concentrations of

lutein are associated with lower risk of coronary heart disease and

stroke.

4.4. Infant nutrition

The two major carotenoids found in human milk are lutein and

zeaxanthin (Giordano and Quadro, 2018). These molecules are crucial

for the visual and cognitive development of infants (Hammond, 2008;

Henriksen & Chan, 2014; Jackson, Lien, White, Bruns, & Kuhlman,

1998). When fed with human milk rather than formula, higher blood

concentrations of lutein/zeaxanthin are found. In a study (Bettler,

Zimmer, Neuringer, & Derusso, 2010) the lutein/zeaxanthin con-

centration increased from 48 µg/L at birth to 96 µg/L at one month in

breast-fed infants. On the other hand, it decreased from 49 µg/L to

33 µg/L in infants under the unfortiﬁed formula. Breastfeeding is es-

sential to avoid lutein/zeaxanthin insuﬃciency during growth. Lutein/

zeaxanthin are the predominant carotenoids found in the infants brain,

making up approximately 66–77% of the total carotenoid concentration

in the brain (Eggersdorfer & Wyss, 2018; Qiao et al., 2009).

5. Lutein content in food sources

Chicken egg yolk has been recognized as an excellent source of lu-

tein. The concentration of lutein in chicken egg yolk is

1622 ± 650 µg/100 g of yolk, and because of its high-fat content, the

bioavailability of lutein is high (Abdel-Aal et al., 2013; Handelman,

Nightingale, Lichtenstein, Schaefer, & Blumberg, 1999; Mangels,

Holden, Beecher, Forman, & Lanza, 1993; Schaeﬀer, Tyczkowski,

Parkhurst, & Hamilton, 1988). Some foods which have been previously

studied for their lutein content are listed in Table 1, including the

processing received before extraction. In some fruits, removal of seeds,

stem, skin, was necessary for better extraction as these parts are non-

edible and do not contribute signiﬁcantly to the total lutein content.

Some foods were processed emulating the conditions which they would

be exposed before consumption; these include, cooking, slicing,

blanching, canning, freezing, and packaging. According to diﬀerent

reports, food processes such as freezing and blanching cause a reduction

of the total lutein content (Fish & Davis, 2003; Morais et al., 2002).

Also, other parameters aﬀecting the lutein content during storage is the

plant matrix itself (species, cultivar), processing (blanching, peeling)

and the storage conditions (temperature, humidity, packaging)

Fig. 2. Structure of all-trans lutein (A). It can be esteriﬁed with fatty acids, including palmitic acid (B), and its oxidation products are usually apocarotenoids (C).

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

Table 1

Lutein content in selected sources.

Source Scientiﬁc Name Content (µg/100 g) Extraction solvent Processing Reference

American gooseberry Pereskia aculeata Mill 250–290 Cold acetone Frozen (Agostini-Costa, Wondraceck, Rocha, & Silva, 2012)

Arazá Eugenia stipitata McVaugh 154–756 Methanol Freeze-dried (Garzón et al., 2012)

Black palm Astrocaryum standleyaum 410–470 Acetone Raw (Murillo, Meléndez-Martínez, & Portugal, 2010)

Broccoli Brassica oleracea 3110–3960 Cold acetone Boiled (De Sá & Rodriguez-Amaya, 2003)

Caryocar villosum fruits Caryocar villosum 70–110 Acetone and petroleum ether/ diethyl ether Freeze-dried pulp (Chisté & Mercadante, 2012)

Cashew Anacardium occidentale 20–40 Acetone Raw (Murillo et al., 2010)

Cilantro Coriander sativum 7703 Methanol Raw (Perry, Rasmussen, & Johnson, 2009)

Guava Psidium guajava L. var. Regional roja 3–11 Methanol/Water Removal of seeds (González, Osorio, Meléndez-Martínez, González-Miret, &

Heredia, 2011)

Jackfruit Artocarpus heterophyllus/Batch A 10.36–55.61 Acetone and petroleum ether/ diethyl ether Pulp extraction (de Faria, de Rosso, & Mercadante, 2009)

Kale Brassica oleracea cv.Manteiga 2860–3500 Cold acetone Stir-fried (De Sá & Rodriguez-Amaya, 2003)

Lettuce Lactuca sativa L. (Mini Romaine - Marta) 1290–1690 Ethyl acetate Raw (López, Javier, Fenoll, Hellín, & Flores, 2014)

Maize Zea mays Amarelao 3 587–593 Hexane/acetone Milled (Kuhnen et al., 2011)

Pepper Capsicum annuum L. (F1 Amanda

hybrid)

670–830 Cold acetone Seeds and stem

removed

(De Azevedo-Meleiro & Rodriguez-Amaya, 2009)

Pitanga Eugenia uniﬂora L. 120–310 Acetone None (Burgos et al., 2012)

Potato Solanum tuberosum subsp. Phureja

Chaucha

135.2–152.4 N,Ndimethylformamide(DMF) Freeze-dried (Fernandez-Orozco, Gallardo-Guerrero, & Hornero-Méndez,

2013)

Potato, Andean Solanum goniocalix (701862) 268–311 Acetone None (Burgos et al., 2012)

Pumpkin (round) Cucurbita máxima 623 Raw (Beltrán, Estévez, Cuadrado, Jiménez, & Alonso, 2012)

Pumpkin, (size squash) Cucurbita maxima 728 Raw (Beltrán et al., 2012)

Sweet potato Ipomoea batatas Lam.. CNPH 1194 1100 Acetone and petroleum ether/diethyl ether Steamed (Donado-Pestana, Salgado, de Oliveira Rios, dos Santos, &

Jablonski, 2012)

Tree tomato Solanum bataceum 120–130 Ethanol/hexane Peeled and ground (Mertz et al., 2009)

White bryony, young shoots Bryonia dioica 19,130 Freeze dried then

milled

(García-Herrera, Sánchez-Mata, Cámara, Tardío, &

Olmedilla-Alonso, 2013)

Marigold ﬂower Tagetes erecta 21,600–97,600 Fresh ﬂowers (Bosma, Dole, & Maness, 2003)

Tagetes erecta 82,900–2,794,600 Dried powder (Delgado-Vargas & Paredes-López, 1996)

Tagetes patula 59,700–1,231,000 Tetrahydrofuran (Manke Natchigal, Oliveira Stringheta, Corrêa Bertoldi, &

Stringheta, 2012)

Calendula oﬃcinalis 4,000–30,100 Tetrahydrofuran, Methanol/ethylacetate/

petroleumether

(Manke Natchigal et al., 2012; Pintea, Bele, Andrei, &

Socaciu, 2003)

Microalgae Chlorella fusca 420,000–470,000 Acetone Dried Powder (Del Campo et al., 2000)

Chlorococcum citroforme 740,000

Tetracysis aplanosporum 590,000

Tetracystis intermedium 350,000

Scenedesmus almeriensis 854,000 Ethanol (Molino et al., 2019)

Desmodesmus protuberans 1,053,000 Hexane/Ethanol (Soares et al., 2019)

Auxenochlorella protothecoides 499,000 Acetone/Methanol Wet biomass (Xiao et al., 2018)

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

(Bouzari, Holstege, & Barrett, 2015). Blanching is a process that re-

quires heat, causing lutein degradation. Aman et al. (2005) reported a

17% loss of total lutein content in blanched spinach compared to

sterilization (26%). The lower temperature and much shorter heat ex-

posure of blanching lead to a lower substantial lutein degradation.

Rubio-Diaz, Santos, Francis, and Rodriguez-Saona (2010) reported that

canning reduces the content of trans-lutein, indicating that heat treat-

ments present strong tendency to isomerize. On the other hand, freezing

temperatures tend to maintain lutein stability during storage. However,

as mentioned by Tacken et al. (2009), refrigeration and freezing has no

eﬀect on the carotenoid and triglyceride content in human milk, except

for lutein. It was shown that the concentrations of Viamin A and E, beta-

carotene and lycopene did not change signiﬁcantly while the lutein

content decreased 24% after 28 days of freezing and 28% after 48 h of

refrigeration. This could be due to the increase in lipid peroxidation at

this temperatures, with lutein being used as an antioxidant (van Zoeren-

Grobben, Moison, Ester, & Berger, 1993).

As seen in Table 1, lutein content from various food sources can be

mainly found in dark green vegetables such as broccoli, lettuce, ci-

lantro, and kale (1290–7703 µg/100 g of plant material) (Walsh et al.,

2015). A decrease in lutein content can be observed in fruits with

higher moisture as is the case with guava, cashew, and jackfruit, con-

taining 3–55.61 µg of lutein/100 g of the source. This may occur due to

the low solubility of carotenoids in water, and/or how the presence of

this high-water content may negatively aﬀect the extraction yield with

organic solvents. Furthermore, the presence of lutein in fruits and ve-

getables can be generally predicted by their color, such as yellow-or-

ange vegetables are found to contain a high amount of carotenoids

(Priyadarshani & Jansz, 2014). In this context, the orange-yellow crops

investigated in Table 1 (pepper, pumpkin, sweet potato) were found to

be good sources of lutein containing 623–1100 µg/100 g plant source.

5.1. Marigold ﬂowers and microalgae

Currently, the primary source of lutein is the marigold ﬂower, a

common name for the genus of Tagetes. Its yellow-orange petals are

abundant in this xanthophyll; it has been reported that as the color of

their petals intensiﬁes, lutein content increases. The most common

colors for lutein production range from orange to deep orange. China

uses T. erecta as the main ﬂower species because of high lutein accu-

mulation of about 20 g/kg found mainly in the ester form (Lin et al.,

2015). As the conventional lutein source, marigold ﬂowers must be

harvested periodically making it a labor-intensive process. The cost

associated with lutein production is high since it is only extracted

during bloom season, occurring annually from July to October.

On the other hand, several microalgae species are potential sources

of lutein as they produce about 5 g/kg biomass mainly in free lutein

form (Table 1). They also present a growth rate that is 5–10 times

higher than plants. Microalgae, which can be cultivated in wastewater

or seawater, do not compete for resources with conventional agri-

culture, making an attractive alternative lutein source (Lin et al., 2015;

Soares, da Costa, Vieira, & Antoniosi Filho, 2019; Yen et al., 2013).

Carotenoids in microalgae are formed via photosynthesis; microalgae

use H

O and sunlight to take up CO

as their primary carbon source.

These microorganisms have a worldwide distribution and are well-

adapted to survive under a broad spectrum of environmental conditions

ranging from icy waters, high osmotic pressure and UV exposure

(Guedes, Amaro, & Malcata, 2011). The main advantages of using mi-

croalgae over marigold ﬂowers are the higher growth rate and lower

water demand. Which implies a high CO

ﬁxation rate, with 1 kg mi-

croalgae being able to store 1.85 kg CO

, importantly reducing its at-

mospheric emission. Even though the lutein content in microalgae is

lower compared to marigold ﬂowers under normal conditions, micro-

algal production of lutein may be combined with other algal-based

metabolites such as vitamins, pigments and polysaccharides (Hsieh-Lo,

Castillo, Ochoa-Becerra, & Mojica, 2019). Microalgae can also be used

for the treatment of wastewaters and the bioﬁxation of CO

.Choix,

Ochoa-Becerra, Hsieh-Lo, Mondragón-Cortez, and Méndez-Acosta

(2018) found Chlorella sp. biomass signiﬁcantly increased when cul-

tured with tequila vinasse with successful ﬁxation of all CO

present.

Coupling these advantages, it is possible to develop a continuous in-

dustrial process that may make microalgae suitable for lutein produc-

tion while bringing environmental beneﬁts. Microalgae can also be

cultivated in tubular bioreactors which can be built vertically, avoiding

the large space of land needed for marigold cultivation (Fábryová et al.,

2019; Molino et al., 2019; Sun, Li, Zhou, & Jiang, 2016; Xiao et al.,

2018). Currently there are no commercial production processes of lu-

tein from microalgae, however, various carotenoid products derived

from microalgae already exist, such as beta-carotene and astaxanthin.

Acién, Fernández, Magán, and Molina (2012) performed production

cost of a real microalgae production plant where they calculated an

annual microalgae production of 70–150 ton/hectare. This was esti-

mated using the microalgae Scenedesmus almeriensi which has a lutein

concentration of 8.54 mg/g. In comparison, annual average production

of fresh marigold ﬂowers (Tagetes patula) is 30–60 tons/hectare with a

lutein concentration of 12.31 mg/g (Lin et al., 2015; Manke Natchigal,

Oliveira Stringheta, Corrêa Bertoldi, & Stringheta, 2012; Molino et al.,

2019). Further studies are needed to overcome the technical obstacles

for microalgae lutein production such as high harvesting cost and high

energy demand for cell disruption and extraction (Lin et al., 2015).

5.2. Extraction from the food matrix

Lutein can be extracted using a variety of organic solvents, such as

acetone, hexane, isopropanol, methanol, diethyl ether, etc. (Table 1). A

variety of solvent combinations could also be used, which provides a

synergistic eﬀect on the yield of extraction of lutein. Consequently, one

of the most critical factors involved in eﬃcient lutein extraction is the

choice of the appropriate solvent or solvent combinations. The carbon

chain length and polarity of the carotenoids, the sample matrix, its

components, and moisture content play essential roles in the selection

of the proper solvent, thus, making the decision rather diﬃcult. Car-

otenoids such as lycopene and β-carotene lack in polar functional

groups, making them highly lipophilic and non-polar. For this reason,

hexane is preferred for the extraction of these carotenoids.

On the other hand, the presence of polar functional groups (epoxy,

hydroxyl, or keto) increases its solubility on solvents like acetone or

ethanol. Because of their water-miscible properties, acetone and

ethanol are preferred for eﬃcient extraction of lutein from plant ma-

terial with high moisture (Amorim-Carrilho et al., 2014; Saini & Keum,

2018). In Table 1, the solvents used for lutein extraction were almost

exclusively of polar nature, also, in the case of maize and tree tomato, a

polar solvent was used in combination with hexane in order to extract

the total content of carotenoids (polar and nonpolar).

The solvents used for extraction present various safety, health, and

environmental risks such as chronic and acute toxicity, skin irritation,

and persistence in the environment. To improve sustainability, more

environmentally friendly solvents and extraction methods should be

explored (Alfonsi et al., 2008). With the purpose on the addition of

lutein in foods, ethanol would be the ideal solvent due to its labeling by

the FDA as a generally recognized as safe (GRAS) substance, which

states that ethanol is safe to use in food products for human con-

sumption (Food and Drug Administration, 2017).

5.3. In vitro studies evaluating the bioaccessibility of lutein

Nutrient bioaccessibility is an important factor regarding the un-

derstanding of the role of dietary components in human health. Interest

in the bioaccessibility of nutrients has increased due to the existence of

undernourished populations (Van Den Berg et al., 2000). This process

can be studied through the application of in vitro and in vivo models. In

vitro models are used to study the pre-absorptive processes and food-

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

matrix related factors aﬀecting carotenoid bioaccessibility

(Chitchumroonchokchai, Schwartz, & Failla, 2004; During, Hussain,

Morel, & Harrison, 2002). However, this should also be studied using in

vivo models.

Furthermore, the mechanism of lutein absorption is not yet fully

understood, but it has been proposed that lutein is absorbed through

the enterocytes by simple diﬀusion or receptor-mediated transport. In

the stomach, lutein is emulsiﬁed into small lipid droplets which are

then incorporated into mixed micelles with the aid of bile salts and

biliary phospholipids; the micelles are then absorbed by enterocytes by

the action of the scavenger receptor class B type I (SR-BI) (Xavier et al.,

2018). Taking this into account, during in vitro gastrointestinal diges-

tion, it is considered that the lutein content present in the micellar

phase reﬂects the amount of lutein available for absorption in the en-

terocyte. Thus, in vitro lutein bioaccessibility from various food sources

depicted in Table 2 was estimated by calculating the proportion of free

lutein in the micellar phase in comparison with the original lutein

content in the food matrix before digestion.

In this context, it is well understood that carotenoid bioaccessibility

is greatly aﬀected by the physicochemical properties of the food matrix

(West & Castenmiller, 1998). For example, in green leafy vegetables,

carotenoids possess signiﬁcant light-harvesting capabilities, they can be

mainly found in the thylakoid membranes of the chloroplasts. On the

other hand, in fruits and roots, carotenoids appear as semicrystalline

structures in the membrane. The diﬀerence in this environment im-

portantly contributes to the extraction methods and analysis needed, as

well as the cellular uptake of carotenoids (Faulks & Southon, 2005). In

2010, a study performed in Spain evaluated the carotenoid bioacces-

sibility in various fruits and vegetables consumed on everyday Spanish

diet (Granado-Lorencio, Herrero-Barbudo, Olmedilla-Alonso, Blanco-

Navarro, & Pérez-Sacristán, 2010). It was found that non-green vege-

tables (tomato paste, red pepper, carrot) showed overall higher bioac-

cessibility of lutein, while green vegetables (broccoli, spinach) showed

the lowest rate; even though they were the most abundant sources of

lutein (Table 2). The authors concluded that this might be associated

with cellular localization of lutein and, as a result, to lutein ex-

tractability. This was further conﬁrmed with the results obtained by

Burgos et al. (2013) (Table 2), which showed that the bioaccessibility of

lutein from yellow-ﬂeshed potatoes was higher (63–71%) than in green

leafy vegetables.

In addition, Xavier et al. (2018) tested the bioaccessibility of lutein

from cupcakes at diﬀerent fortiﬁcation levels (Table 2), ﬁnding an in-

crease of lutein transfer from the product to the micellar phase in for-

tiﬁed cupcakes in comparison with the control. Lutein bioaccessibility

increased from 36% in the control cupcake to 45% and 65% in the 0.5

and 1 mg/serving lutein fortiﬁed cupcakes respectively. Although the

bioaccessibility percentage rises signiﬁcantly, it becomes stable at the

next two fortiﬁcation levels with 61% and 58%, showing very similar

values to the observed in 1 mg lutein/serving. At the next fortiﬁcation

level, the bioaccessibility signiﬁcantly increases reaching 81% at 4 mg

lutein/serving, only to decrease to 60% at 6 mg lutein/serving. It seems

that the correlation between bioaccessibility and fortiﬁcation amount

does not show a linear behavior, meaning that increasing lutein content

per serving does not always result in a net increase in lutein bioacces-

sibility.

Nevertheless, one factor that can aﬀect lutein bioaccessibility is the

inclusion of fats in the meal; this stimulates biliary and pancreatic se-

cretion which can aid in the absorption of lutein. For this reason, the fat

Table 2

In vitro studies evaluating the bioaccessibility of lutein.

Source Model Amount of sample Lutein content in sample Lutein bioaccessibility

(%)

References

Lutein fortiﬁed cupcakes Static gastrointestinal digestion 2 g Control:

0.192-0.196 mg/serving

36 (Xavier et al., 2018)

Fortiﬁed with

0.5 mg/serving

Fortiﬁed with

1 mg/serving

Fortiﬁed with

2 mg/serving

Fortiﬁed with

3 mg/serving

Fortiﬁed with

4 mg/serving

Fortiﬁed with

6 mg/serving

Lutein-fortiﬁed fermented

milk

Static gastrointestinal digestion 5 ml 4 mg/100 ml of milk 95–100 (Granado-Lorencio

et al., 2010)8 mg/100 ml of milk 95–100

Lutein enriched Muﬃn Static gastrointestinal digestion under

fed-state conditions

3 g 36.5 ± 1.37 µg/g 37.9 (Read et al., 2015)

Lutein enriched Cookie 35.2 ± 1.77 µg/g 56.0

Lutein enriched Flatbread 21.5 ± 0.98 µg/g 23.0

Potato (Amarilla Tumbay) Static gastrointestinal digestion 500 mg of freeze-dried an

milled cooked potato

395.7 ± 10.6 µg/g 70.92 (Burgos et al., 2013)

Potato (Amarilla del

Centro)

168.4 ± 4.1 µg/g 63.48

Potato (Ishkupuru) 263.2 ± 13.5 µg/g 70.37

Broccoli Static gastrointestinal digestion 10 g 1305 µg/100 g 7 (Granado-Lorencio

et al., 2007)Spinach 7142 µg/100 g 5

Carrot 354 µg/100 g 14

Tomato paste 83 µg/100 g 92

Red pepper 533 µg/100 g 49

Kiwi 95 µg/100 g 62

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

content of milk makes it an eﬀective delivery vehicle representing a

low-cost alternative to increase micronutrient supply via fortiﬁcation

(Hayes, Pronczuk, & Perlman, 2001). In this respect, fortiﬁed dairy

products are of special interest to populations with unbalanced diets

and micronutrient deﬁciencies (Berner, Clydesdale, & Douglass, 2001).

As presented in Table 2, the total lutein content in lutein-fortiﬁed fer-

mented milk showed a high level of bioaccessibility, with an average of

95 to 100% of the initial content recovered at the micellar phase,

supporting the suitability of fermented milk as an adequate carrier of

lutein for human consumption (Granado-Lorencio et al., 2010).

In this respect, Table 2 also shows lutein bioaccessibility diﬀered

signiﬁcantly between muﬃns, ﬂatbreads, and cookies prepared with

high-carotenoid wholegrain ﬂour, bioaccessibility was lowest for the

ﬂatbread (23.0%), followed by the muﬃn (37.9%), and cookies

(56.0%). These samples contained 0.3%, 12%, and 22% fat respec-

tively, further suggesting the presence of high-fat aids in the bioac-

cessibility of lutein (Read, Wright, & Abdel-Aal, 2015).

5.4. In vivo studies evaluating the bioavailability of lutein

In vivo bioavailability of lutein from various sources is presented in

Table 3.Granado-Lorencio et al. (2010) performed a human study with

twenty-four healthy individuals. They evaluated the bioavailability of

lutein from the consumption of fortiﬁed fermented milk. They found

that the regular consumption of lutein-fortiﬁed fermented milk, at the

level of fortiﬁcation and consumption used (8 mg/day) increased the

serum levels of lutein above the 90 percentile of the reference range in

the US and European populations (> 0.50 μmol/L) (Olmedilla et al.,

2001). In a mouse study, Arunkumar, Prashanth, and Baskaran (2013)

reported that lutein encapsuled polymeric chitosan showed approxi-

mately 2 times greater lutein concentration in plasma, liver, and eye

after 8 h compared to simple emulsion vehicle. In addition,

Vishwanathan, Wilson, and Nicolosi (2009) performed a clinical trial

where they found that a 1 week supplementation of lutein in a stable

hydrophilic nanoemulsion increased plasma lutein concentration 1.3

times compared to lutein delivered in pill form. Because regular dietary

consumption of lutein does not reach those levels associated with its

beneﬁts, it is of great importance to take food formulation and en-

gineering approaches to increase the bioavailability of lutein to yield

signiﬁcant health beneﬁts. In this context, two studies evaluated for-

mulations in order to enhance lutein bioavailability, one utilized na-

noparticle encapsulation (Kamil et al., 2016) and the other employed a

lutein self-emulsifying phospholipid suspension (SEPS) (Shanmugam

et al., 2011) both obtaining positive results. In both cases, increasing

the lutein concentration signiﬁcantly in plasma and retinal tissue (in

the case of SEPS), further encouraging the development of new food

technologies to improve lutein absorption from the food matrix.

6. Food industry processing

The lutein polyene backbone is susceptible to degradation due to the

conditions present during food processing which may result in the

fragmentation of the molecule. Energy in the form of light, heat and

mechanical stress are capable of interrupting conjugation of the mole-

cule causing loss of color and biological activity (Martínez-Delgado

et al., 2017). Nowadays the production of commercial lutein involves

four steps. The cultivation, pretreatment, processing and ﬁne proces-

sing. Fresh Tagetes ﬂowers are currently the raw material used for lutein

Table 3

In vivo studies evaluating the bioavailability of lutein.

Source Model Form of

administration

Lutein dose Bioavailability References

Lutein-fortiﬁed fermented milk Men and women,

18–30 years apparently

healthy

Oral Basal plasma level. 0.009 µmol/L (Granado-Lorencio

et al., 2010)200 ml.

Total dose supplied:

8 mg

0.163 µmol/L

200 ml.

Total dose supplied:

16 mg

0.560 µmol/L

PLGA nanoparticles

(Polylactic-co-glycolic acid)

Rats.

Male (Fischer 344) Body

weight 238 ± 8.0 g

Gastric gavage 10 mg/kg BW

Free Lutein (~2.38 mg/rat)

90.2 ± 18.1 ng/mL plasma (Kamil et al., 2016)

10 mg/kg BW

PLGA Lutein (~2.38 mg/rat) 1.7 ± 1.4 ng/mL

plasma

Lutein self-emulsifying

phospholipid suspension

(SEPS)

Beagle dogs Oral 100 mg lutein. Commercial

formulation

23.63 ± 19.28 ng/mL

plasma

(Shanmugam et al.,

2011)

100 mg lutein. SEPS 277.93 ± 92.67 ng/mL

plasma

Sprague Dawley rats 100 mg/kg/day dose of

lutein for 14 days.

Commercial formulation

3.45 ± 1.63 ng/g

Retinal lutein content

100 mg/kg/day dose of

lutein for 14 days.

SEPS

14.72 ± 2.02 ng/g

Retinal lutein content

0 mg/kg/day

Placebo

0.91 ± 0.31 ng/g Retinal lutein

content

Broccoli Men and women,

20–35 years apparently

healthy

Orally at lunch or

dinner.

200 g broccoli/day

for 7 days

(1373 µg lutein/ 100 g

broccoli)

0.30 µM in serum (Granado et al.,

2006)-

Basal Lutein content. (start of

the experiment

0.20 µM in serum

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

extraction, they are harvested and then sent to processing facilities.

Flowers are ensiled (anaerobic fermentation) with the aim of preserving

them for longer periods of time during storage. After ensilage, only the

petals are dried, crushed and compressed to make the “marigold

granules”, which are considered the ﬁrst crude lutein product. These

granules must pass through an extraction process to make a second

product: “marigold oleoresin” or “lutein oleoresin”. Finally, if the lutein

products require higher purity, it can be obtained through ﬁnal process

saponiﬁcation and puriﬁcation of the “marigold oleoresin” (Lin et al.,

2015). Their hydrophobic nature has conﬁned carotenoid almost en-

tirely to lipid systems (emulsions); large amounts of carotenoids can be

present in very ﬁne dispersions capable of coloring aqueous matrices.

The use of carotenoids for coloring lipid phase food matrices can be

achieved easily. Products such as oils, lards, dressings, margarine and

butter are frequently colored using carotenoids. Usually, after the fatty

product has been clariﬁed, the carotenoid suspension is added to the

warm product (40–50 °C) with agitation until the complete dissolution

is achieved. Subsequent treatments as chilling are performed. If anti-

oxidants are added for the protection of the oxidative rancidity of the

fat, they will also provide an added stabilizing inﬂuence on the car-

otenoids. In bakery use, the color carried to cookies, cake frosting,

doughnuts, etc. will depend on temperature and color level. High

temperature process, such as deep-frying oils may aﬀect carotenoid

stability. On-going research in carotenoid chemistry and formulation

will improve and develop new manufacturing methods that could in-

crease the its application in food matrices and expand the market with

broader applications (Kruger, Murphy, DeFreitas, Pfannkuch, &

Heimbach, 2002; Mortensen, 2006; Rodriguez-Amaya, 2016). In plant

cells, carotenoids are mostly stored inside of the chloroplast, protecting

them from external contact with other cell components. Once cellular

integrity is lost, carotenoids are prone to degradation due to environ-

mental stress. As shown (Table 4) during processing of carrot and blood

orange juice, where lutein degradation is in a range of 26–50%, because

during juice extraction, cellular integrity was lost providing minor re-

sistance and causing a major degradation (Chen, Hsieh, Lee, Chang, &

Chang, 2016). Also, due to the low water-solubility of lutein, fat-rich

formulations (emulsion-based systems) may reduce the degradation in

the food matrix by limiting the presence of hydrophilic free radicals

(Meléndez-Martínez, Vicario, & Heredia, 2004). Due to the low water-

solubility of lutein, fat-rich formulations (emulsion-based systems) may

reduce the degradation in the food matrix by limiting the presence of

hydrophilic free radicals (Meléndez-Martínez et al., 2004). In Table 4

can be observed that 26–50% of lutein is lost during processing of

carrot and blood orange juice. In these cases, cellular integrity was lost

during juice extraction providing minor resistance to degradation (Chen

et al., 2016).

6.1. Eﬀect of temperature

During food processing, particularly thermal processing (e.g.

blanching, pasteurization, cooking, canning, frying, and drying) lutein

concentration may decrease, but at the same time, it may be beneﬁcial

through the disruption of cell walls and membranes (Maiani et al.,

2009). The inﬂuence of temperature on lutein concentration is shown in

Table 4. High temperatures accelerate the rate of the degradation re-

action of carotenoids (Abdel-Aal et al., 2010). In general, lutein con-

tains all its double bonds in the form of the trans isomer, which is

partially transformed into the cis form during thermal exposure, this

form is thermodynamically less stable than the trans isomer (Meléndez-

Martínez et al., 2004). Since thermal degradation is a common phe-

nomenon in lutein, it is critical for maintaining it at the safe tempera-

ture range during food processing. Heat treatment to which the food

matrix is subjected, especially in the presence of oxygen, can lead to

isomerization and degradation of lutein (Boon et al., 2010). Ahmad,

Asenstorfer, Soriano, and Mares (2013) concluded that the degradation

of lutein due to diﬀerent heating conditions follows ﬁrst-order kinetics.

When the temperature reaches 40 °C, the overall lutein loss was low

over the storage period used in their study. At temperatures between 50

and 60 °C lutein loss increases signiﬁcantly, but when stored at high

temperature (≥80 °C), losses of lutein are signiﬁcant.

6.2. Eﬀect of UV light

The strong presence of light can induce the breakage of carotenoids,

resulting in the formation of low molecular weight compounds. Kline,

Duncan, Bianchi, Eigel, and O’Keefe (2011) reported that the most

damaging wavelengths to lutein were found to be in the UV range of

200–400 nm and 463 nm. This agrees with the results from Khalil et al.

(2012) (Table 4), they reported lutein degradation up to 55% in 365 nm

for 72 h. However, it was found that emulsiﬁcation of lutein in medium-

chain triacylglycerols (MCT) oil improved the stability of lutein ester

extract against UV light at 365 nm. This underlines the fact that besides

thermal stability, UV stability should also be given for long-term sto-

rage and usage of fortiﬁed foodstuﬀ.

6.3. Oxidation

The most common cause of lutein degradation is due to oxidation,

usually occurring via chemical or enzymatic reactions (e.g., lipox-

ygenases) most commonly during the drying process of fruits and ve-

getables. In a food matrix, the lutein oxidation mechanism is very

complex due to its dependence on many factors. Lutein may go through

auto-oxidation when there is exposure to atmospheric oxygen, the rate

Table 4

Condition and matrix eﬀect on lutein stability.

Process Conditions Source Lutein loss (%) References

Light exposure 25 °C, 2550 Lx/72 h Lutein bovine casein emulsions 5 (Mora-Gutierrez et al., 2018)

Ultrasonic waves 80 kHz, 240 W, 30 °C, 2.5 h Ethanolic lutein solution 12 (Song, Li, Pang, & Liu, 2015)

Pasteurization/ Cheese-making 63 °C/30 min

pH 5.2

Rinsing with 80 °C water

Mozzarella cheese 20 (Liu, Wang, Liu, & Ren, 2018)

Temperature Oil bath 75 °C/150 min Blood orange juice 26 (Hadjal, Dhuique-Mayer, Madani, Dornier, &

Achir, 2013)

Pasteurization 110 °C/30 s Carrot juice 30 (Chen, Peng, & Chen, 1995)

Canning 120 °C/30 s Carrot juice 50 (Chen et al., 1995)

UV light UV light (365 nm; 12 W)/

48 h

Lutein orange-oil emulsion from Tagetes

patula

55 (Khalil et al., 2012)

Microencapsulation 40 °C, 75% RH, 20 days Spray-dried lutein microcapsules 60 (Álvarez-Henao et al., 2018)

Temperature 100 °C/40 min Lutein orange-oil emulsion from Tagetes

patula

61 (Khalil et al., 2012)

Storage under refrigeration 4 °C/60 days Lutein-loaded lipid-core nanocapsules 64 (Brum et al., 2017)

Temperature 60 °C/20 days Wheat grain 80 (Ahmad et al., 2013)

M. Ochoa Becerra, et al. Journal of Functional Foods 66 (2020) 103771

of which oxidation occurs will change depending on the presence of

light, high-temperature, pro-oxidants, and antioxidants. The oxidative

process brings the formation of apocarotenoids, carotenoids containing

less than 40 carbon atoms (Fig. 2C), and low molecular weight com-

pounds similar to the ones found after fatty acid oxidation. As a result,

pigmentation and bioactive properties are lost (Meléndez-Martínez

et al., 2004).

7. Preventing lutein degradation in a food matrix

Interaction of lutein with other food constituents can exert protec-

tion against oxidation reactions, for instance by adding a higher con-

centration of saturated fat sources that are less likely to be oxidized,

limiting the presence of free radicals (Meléndez-Martínez et al., 2004).

Other alternatives are the use of opaque and airtight containers that

help to avoid contact with oxygen and light. Furthermore, freezing or

refrigeration, the inclusion of antioxidants and a nitrogen-rich en-

vironment can reduce losses of the pigments (Boon et al., 2010).

In this regard, new technologies are currently being investigated in

an attempt to increase the stability of carotenoids (Steiner et al., 2018).

Microencapsulation can reduce the oxidation of bioactive compounds

present in foods and can easily allow for better powder dispersion in

water (Rigon & Zapata Noreña, 2016). Nanoemulsion and nanocapsules

are other techniques used to stabilize lutein, a particle size of less than

1 µm help in lutein solubility, thus improving bioavailability and in-

creasing stability (Brum et al., 2017) Chitosan nanoparticles has been

previously developed to enhance solubility of lutein, showing an in-

crease of 58% on stability compared to the polyethylene glycol nano-

capsules. Suggesting that nanoencapsulation is an eﬃcient carrier of

lipophilic molecules such as carotenoids. Boon et al. (2010) reported

that multilayer emulsions around the oil droplets could potentially re-

duce the amount of light that could reach carotenoids depending on the

thickness of the membrane.

8. Concluding remarks

The search for high lutein sources has led to the utilization of

marigold ﬂowers as the primary source for industrial production.

However, ﬂower harvesting is considered to be very labor-intensive,

and because they present an annual growth, lutein can only be ex-

tracted periodically. Microalgae have been recognized as a possible

alternative for lutein production, as they could capture CO

emissions

and can be harvested all year. In addition, lutein health-promoting

properties have led to its inclusion as a functional food ingredient. The

compatibility of lutein with other food components and exposure to

certain environmental factors such as high temperatures, the presence

of light and oxygen, may cause the degradation of the molecule.

Promising stabilization techniques such as nanoemulsions and micro-

encapsulation should be more widely explored. They have shown to

increase the lutein stability and bioavailability without compromising

its biological activity and pigmentation capacity. However, the lack of

information in vivo models requires further investigations to understand

the absorption, metabolism pathway and action mechanism of lutein in

humans.

On the other hand, bioaccessibility and bioavailability of lutein

from the same matrix are diﬀerentially aﬀected by the intrinsic prop-

erties of the food matrix (ripening state), processing (thermal or me-

chanical), and the presence of some dietary components such as fats,

ﬁber or phytosterols. All these factors do not act on their own, but they

interact with each other, interactions which should be further studied.

9. Ethics statements

This is a review paper, which doesn’t include animal or human

experiments.

Declaration of Competing Interest

The authors declare that they have no known competing ﬁnancial

interests or personal relationships that could have appeared to inﬂu-

ence the work reported in this paper.

Acknowledgments

I would like to express my very great appreciation to Cecilia Moreno

for her valuable and constructive suggestions, proofreading and advice

during the development of this work.

Author Mario Ochoa was supported by a scholarship from the

Consejo Nacional de Ciencia y Tecnología CONACyT-México, number

836478.

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Genetic assessment and trait profiling of marigold (Tagetes spp.) hybrids

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Jan 2025

Marigold (Tagetes spp.) is a highly valued loose flower crop of the floriculture industry, prized for its versatility, colourful blooms and ease of cultivation. Its resilience, low maintenance and adaptability make it an ideal choice for farmers, offering an excellent opportunity for profitable farming. Additionally, marigolds are rich in carotenoid content and have several pharmaceutical uses. This study evaluates genetic variability among key morphological traits including flower yield in 20 marigold hybrids. ANOVA analysis revealed significant variations across all traits, indicating considerable diversity within the genotypes. The phenotypic coefficient of variation was higher than the genotypic coefficient of variation for all traits, suggesting a notable environmental influence on trait expression, although genetic factors still played a dominant role due to very small differences. Heritability estimates were very high, ranging from 79.55% (plant spread) to 99.35% (flower weight). High heritability along with high genetic advance was observed for ray florets per flower, flower weight and yield per plant, indicating that selection for these traits is likely to result in substantial genetic improvement. Principal component analysis (PCA) identified four principal components, which together accounted for 83.57% of the total variability. Flower size, ray florets per flower, plant height and flower count per plant were the primary contributors to the four principal components, respectively.

Gas chromatography-mass spectrometry characterization of carotenoids produced by yeasts isolated from edible fruits and vegetables marketed in Osogbo metropolis, Southwest Nigeria

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Apr 2025

Globally, there is a rising demand for the replacement of chemically synthesized carotenoids (which dominate the market and negatively impact the environment and public health) with organic carotenoids. Therefore, we produced and characterized carotenoids from yeasts isolated from edible fruits and vegetables from three local markets in Osogbo, Nigeria. Five samples each of fresh edible fruits (orange, banana, pineapple, watermelon, and mango) and vegetables (carrot, cucumber, lettuce, green pepper, and tomato) were collected from Shasha, Testing Ground, and Oke-Baale markets. Yeasts were isolated on plate count agar (PCA) and presumptively identified using standard methods. Pigmented colonies were carefully picked, purified, and presumptively identified. Mass cultivation of pure culture was done on plate count agar, and extraction was done using chloroform after cell hydrolysis. The extracts were characterized using gas chromatography-mass spectrometry (GC–MS), and the yeast isolates' identity was confirmed using molecular methods. A total of 24 yeasts were obtained from the fruit and vegetable samples. Four pigmented yeasts, Rhodotorula mucilaginosa (LETA) and (LETB), Rhodotorula toruloides (GP-1), and Candida haemulonis (GPS-1) were obtained from lettuce and green pepper samples, respectively. The GC–MS analysis revealed the presence of 35 compounds in pigments from LETA and LETB, 20 compounds from GP-1, and 33 compounds from GPS-1. The molecular ion (m/z) values 441 m/z, 398 m/z, 486 m/z and 337 m/z recorded in pigment LETA, LETB, GP-1, and GPS-1 suggest the presence of β-carotene, lycopene, lutein, and phytoene, respectively. The identity of the yeasts was confirmed, and accession numbers PPO78657.1, PPO78678.1, PPO78659.1, and PPO78660.1 were assigned correspondingly. This study unveiled the pigmented yeasts from lettuce and green pepper samples as a viable alternative source of carotenoids. Further studies on the purification and application of carotenoids are recommended.

Utilization of Wheat with Enhanced Carotenoid Levels and Various Fat Sources in Hen Diets

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Full-text available

Apr 2025

In this study, we evaluated the effects of two wheat varieties with different carotenoid concentrations (Pexeso and Tercie) and two fat sources with contrasting fatty acid profiles (rapeseed oil and lard) on hen performance, egg quality, and egg yolk retention of carotenoids. The feed conversion ratio of hens that received Tercie wheat and lard in their diet were greater than those of hens that received other diets (p = 0.002). Greater (p ˂ 0.001) eggshell thickness and strength occurred when hens were fed a Pexeso wheat diet. Moreover, Pexeso wheat and lard increased lutein (p ˂ 0.001 and p = 0.001) and zeaxanthin (p ˂ 0.001 and p = 0.001) contents in egg yolks. The highest lutein retention (p = 0.010) occurred in the groups that received Pexeso wheat (46.4 and 47.4%), and the highest zeaxanthin retention (p = 0.011) occurred with a Pexeso wheat and lard diet (59.5%). The lowest lutein and zeaxanthin retention occurred in hens fed a Tercie wheat and rapeseed oil diet (23.6% for lutein retention and 24.1% for zeaxanthin retention). The Pexeso wheat and rapeseed oil diet increased the concentrations of α- and γ-tocopherol (p ˂ 0.001 and p ˂ 0.001) in egg yolks, which influenced the oxidative stability of the eggs. Compared with other diets, a Tercie wheat and rapeseed oil diet led to the lowest oxidative stability in fresh eggs (p = 0.041). In conclusion, Pexeso wheat had greater retention of biologically active substances and higher mineral contents than Tercie wheat, which was reflected in the performance of hens and the quality of eggs. The combination of Pexeso wheat with rapeseed oil, which is rich in tocopherols and polyunsaturated fatty acids and has a favorable n-6/n-3 ratio, increased the tocopherol content and the oxidative stability of egg yolk fats.

A sustainable and efficient method for sequential extraction of lutein and lipid from deep eutectic solvent pretreated Chlorella pyrenoidosa

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Full-text available

Apr 2025

Microalgae biomass is regarded as a potential feedstock for valuable compounds such as pigments, lipids and proteins. However, development of single molecule extraction processes is the most common practice. A green multiproduct extraction approach is needed for economically sustainable process development of the microalgal industry. Therefore, this study aims to investigate the sequential extraction of lutein and lipid from dry and wet Chlorella pyrenoidosa biomass pretreated with a choline chloride-based deep eutectic solvent (DES) under a sustainable biorefinery scheme. In this context, we have assessed the kinetic modeling of the solid–liquid extraction process for the aforementioned compounds, focusing on the effects of temperature and time. The maximum lutein (3.80 mg g⁻¹) and lipid (95.0 mg g⁻¹) contents from dry biomass were obtained at 45 °C in 40 min and at 70 °C in 90 min, respectively. From wet biomass, the maximum lutein (2.57 mg g⁻¹) and lipid contents (87.47 mg g⁻¹) were obtained at 35 °C in 40 min and at 70 °C in 90 min, respectively. The kinetics of the solvent-based extraction process for lutein and lipids were assessed via first-order and second-order kinetic models with an associated investigation of kinetic parameters, such as rate constants, saturation concentration and activation energies. We found that temperature is an important parameter that influences the extraction of all compounds and also has a significant impact on the kinetic parameters. Toxicity evaluation of the DES and economic assessment of DES vs. ionic liquids (ILs) were performed. The synthesis cost of the DES is lower than that of ILs, and Escherichia coli JM109 survivability assessment confirms the DES as a non-toxic solvent. The present study provides valuable insights into the sequential extraction for a high-value multiproduct biorefinery.

Biological Activities of Lutein

Chapter

Apr 2025

Phytochemical profiling of Tagetes erecta L. flowers at various blooming stages through optimized extraction of bioactive compounds for the development of functional juice

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Full-text available

Apr 2025

Introduction Tagetes erecta L. is a valuable medicinal plant and its edible flowers are a rich source of secondary metabolites that may vary at different blooming stages. Objective Current study aimed to evaluate the phytochemical profile of flowers at different blooming stages with varying treatments. Methodology Extracts were prepared by employing solvent extraction with ultra-sonification as pretreatment and by varying solvents to assess the phenolic and antioxidant activities. The high-performance liquid chromatography analysis was performed to quantify bioactive polyphenolics to validate effect of various stages and treatments on sample extracts. Results Study found that ultrasound-pretreated samples resulted higher yield and phenolic contents than solvent extraction (CSE) alone. The highest carotenoid content (427.39 mg/100 g) was observed for the pretreated full-bloom flower samples (MAF-1). Radical scavenging activities were observed highest (DPPH 96.19% and ABTS* 160.09 mmol TE/100 g) in Tagetes sample MAF-1 followed by MEF-1 and MAI-1. A highly significant and positive correlation was observed among all variables of phenolic and antioxidant assays. Various bioactive compounds including phenolic acids, flavonoids and carotenoids were quantified by HPLC-DAD. Results showed that the ultra-sonified extracts (MAF-1) resulted in higher gallic acid (109 mg/g), quercetin (11.74 mg/g), lutein (1.76 mg/g) and zeaxanthin (2.14 mg/g) followed by MEF-1 > MAI-1 > MEI-1. Conclusion The study results showed that the ultrasound-assisted extracts resulted in higher bioactive contents and antioxidant activities expected to deliver more value as additive to functional foods. It also reflects the potential of Tagetes flowers for the functional foods aiming to control early age-related disorders (ARDS).

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Apr 2025
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Bottlenecks in applications of algal-based technologies toward industrial commercialization

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Jan 2025

Meat Products Enriched with Antioxidants

Chapter

Mar 2025

In recent years, there has been a growing trend in the food industry towards enhancing the nutritional profile and quality of meat products. One innovative approach involves enriching meat products with antioxidants. These antioxidants are substances that can counteract the damaging effects of free radicals in the body, which are associated with various health issues, including cell damage and chronic diseases. When incorporated into meat products, antioxidants have the potential to not only improve the product’s shelf life but also offer health benefits to consumers. The impact of temperature on the food matrix during industrial food processing is significant, especially for heat-sensitive chemicals. Therefore, antioxidants found in oilseeds, grains, legumes, fruits, vegetables, herbs, and spices, and other food sources are also receiving attention. This review explores the potential benefits, challenges, and future directions of antioxidant-enriched meat products in the context of consumer health and dietary choices.

Bench-Scale Cultivation of Microalgae Scenedesmus almeriensis for CO2 Capture and Lutein Production

Article

Full-text available

Jul 2019

In this study, Scenedesmus almeriensis as green microalga was cultivated on bench-scale for carbon dioxide (CO2) capture and lutein production. The autotrophic cultivation of S. almeriensis was carried out by using a vertical bubble column photo-bioreactor (VBC-PBR) with a continuous flow of a gaseous mixture of oxygen (O2), nitrogen (N2), and CO2, the latter in content of 0.0–3.0 %v/v. The liquid phase was batch. S. almeriensis growth was optimized. In addition, lutein extraction was carried out by using accelerated solvent extraction with ethanol as Generally Recognized as Safe (GRAS) solvent at 67 °C and 10 MPa. Upon optimization of CO2 concentration, the maximum biomass productivity, equal to 129.24 mg·L⁻¹·d⁻¹, was achieved during the cultivation by using a content of CO2 equal to 3.0 %v/v and it allowed to obtain a lutein content of 8.54 mg·g⁻¹, which was 5.6-fold higher in comparison to the analogous process carried out without CO2 addition. The ion chemical analysis in the growth medium showed that by gradually increasing CO2 content, the nutrient consumption during the growth phase also increased. This study may be of potential interest for lutein extraction at industrial scale, since it is focused on pigment production from a natural source with a concomitantly CO2 capture.

Analysis of major carotenoids and fatty acid composition of freshwater microalgae

Article

Full-text available

Apr 2019

Considering the nutraceutical properties, the high commercial value from pigments and essential lipids and the environmental sustainability, the purposes of this study were to assess the major carotenoids and fatty acids composition of nine microalgae species as a source of nutraceutical compounds and as fatty raw material for biodiesel production. The carotenoid and fatty acid content were analyzed by high performance liquid chromatography tandem mass spectrometry detection method with atmospheric pressure chemical ionization mode (HPLC/APCI-MS/MS) and by high resolution gas chromatography with flame ionization detector (GC-FID). For the carotenoid analysis, the developed method presented a rapid response, a good chromatographic separation, higher sensitivity and can provides more compounds information due the mass spectrum. Among the microalgae evaluated, Desmodesmus protuberans (10.3 mg g ⁻¹ ), Desmodesmus denticulatus var. linearis (8.43 mg g ⁻¹ ) and Chlamydomonas planctogloea (7.4 mg g ⁻¹ ) are good lutein sources. Coelastrum sphaericum (15.29 mg g ⁻¹ ) and Parachlorella kessleri (22.96 mg g ⁻¹ ) showed high astaxanthin content; the others microalgae species presents low carotenoid content. In addition, Chlorella zofingiensis provides high quantities of γ-linolenic acid (4.3%). Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels were lower than 1.1 %. Regarding for biodiesel production, the promising strains are Coelastrum sphaericum and Parachlorella kessleri.

Photosynthetic Accumulation of Lutein in Auxenochlorella protothecoides after Heterotrophic Growth

Article

Full-text available

Aug 2018
MAR DRUGS

In order to enhance lutein accumulation and to explain the reasons for the difference in lutein accumulation under photoautotrophic and heterotrophic conditions, different culture modes and the associated transcriptome profiles were investigated in Auxenochlorella protothecoides. The heterotrophic-photoautotrophic transition culture mode was investigated for lutein accumulation, changing from organic carbon to increase biomass in dark fermentation to irradiation under nitrogen rich conditions. This strategy increased the lutein content 10 times along with chloroplast regeneration and little biomass loss in 48 h. The highest lutein productivity and production in the heterotrophic-photoautotrophic transition culture reached 12.36 mg/L/day and 34.13 mg/L respectively within seven days. Furthermore, compared to the photoautotrophic conditions, most genes involved in lutein biosynthesis and photosystem generation were down-regulated during heterotrophic growth. By contrast, two β-ring hydroxylases were transiently upregulated, while violaxanthin de-epoxidase and zeaxanthin epoxidase were mostly downregulated, which explained the extremely low lutein content of heterotrophic cells. Nevertheless, the lutein proportion in total carotenoids reached nearly 100%. This study is the first to our knowledge to report on a comparative transcriptome analysis of lutein biosynthesis, and it provides a promising strategy to boost lutein production in A. protothecoides.

Phycocyanin and phycoerythrin: Strategies to improve production yield and chemical stability

Article

Sep 2019

Phycocyanin and phycoerythrin are photosynthetic pigments extracted from microalgae with great biotechnological potential due to their intense colors, fluorescent properties and potential health benefits. Their principal applications are as nutritional supplements, natural colorants in foods and cosmetics, and as a reagent for immunological assays. However, the main challenges for their commercialization and implementation in food and cosmetic applications are their low yield during production and limited chemical instability. This review compares the use of different methods to increase biomass and pigment production yield, including the mechanisms associated with the microorganisms adaptative response to different light wavelengths. It also explores different methods reported to improve phycocyanin and phycoerythrin chemical stability and commercially available products.

Purification of lutein from the green microalgae Chlorella vulgaris by integrated use of a new extraction protocol and a multi-injection high performance counter-current chromatography (HPCCC)

Article

Jun 2019

Lutein is a natural carotenoid with health promoting properties whose commercial source is the yellow Marigold flower, but its production is limited by climate, planting area and high labor costs. The microalga Chlorella vulgaris is a promising lutein source; however, no commercial lutein production uses microalgae. In the present study, the lower phase (LP4) of the biphasic solvent system composed of n-heptane–ethanol–water (5:4:1.5, v/v/v) was used both as a solvent for microalgae biomass extraction and as a mobile phase for lutein isolation using high performance counter-current chromatography (HPCCC). The ultrasound-assisted extraction of biomass with LP4 for 30 min led to an extract with a lutein content (3.20 mg/g dried biomass) that was higher than those obtained by the reference solvents tetrahydrofuran and dichloromethane. A multi-injection HPCCC method was developed for lutein isolation, where LP4 was pumped as the mobile phase at a flow rate of 8 mL/min, whereas its upper phase was the stationary phase. In total, 2 g of C. vulgaris extract were processed affording 60 mg of lutein (92% purity), which was further cleaned up by gel permeation chromatography yielding 50 mg of lutein (97% purity). The chemical identity of the purified compound was confirmed by mass spectrometry and absorption spectroscopy in comparison with an authentic standard. The developed HPCCC method may serve as a reference for promoting the development of a new production model of microalgae-based lutein

Encapsulation systems for lutein: A review

Article

Oct 2018
TRENDS FOOD SCI TECH

Background: The increased demand by consumers for clean labels has encouraged industry to search for replacements of synthetic ingredients in food products, and in particular, colorants. Lutein, a xanthophyll found in marigolds and corn, can be used in food products as a natural colorant replacing yellow food dyes. Moreover, lutein is considered a nutraceutical due to its potentially beneficial health effects, such as prevention of macular degeneration, role in the development of the visual and nervous systems of fetuses, and its antioxidant properties. However, incorporation of lutein into foods is often limited because of its low-water solubility, chemical instability, and poor oral bioavailability. For this reason, colloidal encapsulation systems have been developed to facilitate the incorporation of lutein into aqueous food and beverage products. Scope and approach: This review focuses on exploring encapsulation options for lutein using various emulsion-based, nanoparticle- and microparticle-based and molecular inclusion encapsulation systems, as well as additives that can be used to increase its chemical stability in these systems. This review covers all aspects of lutein encapsulation, including both food-grade and pharmaceutical-grade encapsulation systems. Key findings and conclusions: Though lutein-loaded encapsulation systems are extensively explored in this review, emulsions are of the most interest in industry as they are cost efficient and can be designed to increase the stability of lutein by selecting the proper emulsifiers and emulsification techniques. Despite the extensive amount of research carried out on the encapsulation of hydrophobic bioactive molecules such as lutein, there are still opportunities to develop encapsulation systems that further protect these molecules during storage and also increase their bioavailability after ingestion.

Fate of lutein-containing zein nanoparticles following simulated gastric and intestinal digestion

Article

Aug 2018
FOOD HYDROCOLLOID

Zein nanoparticles (ZNPs) were prepared to encapsulate lutein via solvent-induced nanoprecipitation, and the stability of the zein nanoparticles with encapsulated lutein (ZLNPs) was determined following in vitro gastric and intestinal simulation. Stability was assessed by dynamic light scattering, gel electrophoresis, tendency towards sedimentation, and both atomic force and light microscopy. ZNPs possessed a hydrodynamic radius of ∼75 nm, which was not altered with incorporation of lutein. Gastric digestion conditions induced significant aggregation and sedimentation of ZLNPs, which were not fully digested by gastric enzymes and were found adhered to lipid droplets in light micrographs. Aggregation was decreased and digestion was promoted during gastric digestion if salt was omitted, indicating that a high ion concentration increased ZLNP aggregation and limited enzymatic digestion. ZLNPs were redispersed in intestinal conditions and completely digested into peptides. In comparison to aqueous lutein dispersions, incorporation within ZNPs increased lutein's digestive stability by ∼58% but reduced its micellarization efficiency by ∼42%. These findings indicated that ZNPs provided a degree of physical protection to encapsulated carotenoids in gastric conditions yet might partially interfere with certain pathways for carotenoid bioaccessibility.

Carotenoids in human nutrition and health

Article

Jun 2018

Carotenoids are naturally occurring pigments found in most fruits and vegetables, plants, algae, and photosynthetic bacteria. Humans cannot synthesize carotenoids and must ingest them in food or via supplementation. Carotenoids have a range of functions in human health. They primarily exert antioxidant effects, but individual carotenoids may also act through other mechanisms; for example, β-carotene has a pro-vitamin A function, while lutein/zeaxanthin constitute macular pigment in the eye. The benefit of lutein in reducing progression of age-related macular eye disease and cataracts is strengthening; an intake recommendation would help to generate awareness in the general population to have an adequate intake of lutein rich foods. There is evidence that carotenoids, in addition to beneficial effects on eye health, also produce improvements in cognitive function and cardiovascular health, and may help to prevent some types of cancer. Despite the evidence for the health benefits of carotenoids, large population-based supplementation studies have produced mixed results for some of the carotenoids. To establish and confirm the health benefits of the different carotenoids more research, including clinical studies, is needed.

Development of mass transfer model for chromatographic separation of free lutein and fatty acids in de - esterified marigold lutein

Article

Apr 2018
FOOD BIOPROD PROCESS

This article was aimed to construct a mathematical mass transfer model for chromatographic separation of free lutein and fatty acids in de-esterified marigold lutein. Required model parameters: adsorption isotherm, overall mass transfer and axial dispersion coefficients, were first determined. The adsorption isotherms of the compounds were studied using batch adsorption experiments, while the axial dispersion and overall mass transfer coefficients were determined from published empirical-correlation and correlation developed experimentally from this study, respectively. The adsorption isotherms were found to be linear within the equilibrium concentration of ≤100 μg/ml. The isotherm constants of fatty acids were about twice those of free lutein. The isotherm constants and the mass transport parameters were then applied to the mass transfer model. The predicted results were compared with the experimental data at the optimum mobile phase velocity (0.16 cm/s). Good agreement was found, i.e., the average absolute deviations were less than 2% and 7% for semi-preparative and preparative columns, respectively.

Lutein, zeaxanthin and mammalian development: Metabolism, functions and implications for health

Article