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Systemic levels of carotenoids from mangoes and papaya consumed in three forms (juice, fresh and dry slice)



Vitamin A deficiency is a public health problem in Cameroon. Data on the bioavailability of carotenoid in fruits currently consumed in Cameroon are scarce.


To assess the systemic levels of carotenoids from mangoes and papaya consumed as juice, fresh or dried slices.


Two groups of seven healthy volunteers (24 and 25 years of age; body mass index: 21 and 22 kg/m2 respectively for subjects fed mango and papaya), were submitted to three types of meal treatments (juice, fresh and dried fruit). On the experiment day, meals served to fasting subjects during breakfast, included bread, yogurt and one of the three forms of fruit. All the treatments lasted only one day during which blood samples were collected three times; during fasting (T0), 4 h (T4) and 8 h (T8) after the test meal. The carotenoids and retinol contents were analysed by high-performance liquid chromatography method.


From the major carotenoids present in papaya and mangoes, lutein, α-carotene and β-carotene were found in considerable amounts. Lycopene and cryptoxanthin that were the major carotenoids in papaya samples appeared in low amounts in the chylomicrons. Significant correlations were observed between these carotenoids (at T0, T4 and T8). The three forms of consumption contributed to the rise of serum retinol levels. A comparison between the three forms revealed that papaya and mangoes consumed in form of juice or fresh fruit are the best forms because they had higher bioavailability values.


Association of these different forms of consumptions could lead to a better availability of these fruits throughout the year and therefore efficiently contribute to improve vitamin A status of the population.


Provitamin A carotenoids constitute an important source of vitamin A in developing countries. They are mainly derived from dark-green leafy vegetables, yellow/orange fruits, yellow roots tubers and red palm oil (FAO/WHO, 1998). The predominant provitamin A carotenoids found in yellow/orange fruits are β-carotene and β-cryptoxanthin (Kimura et al., 1991). Apart from provitamin A carotenoids, fruits contain other carotenoids such as lutein as well as lycopene, which are associated with the protection of the retina macula from degeneration and reduced risk of developing certain types of cancers in human (Block et al., 1992; Giovannucci, 1999).

In rural areas of Cameroon, fruits are either produced in farmlands or picked from wild trees and consumed in fresh form within short periods of seasonal availability. Industrial processing of surplus fruits for off-season use is not well developed and most often, transportation to non-producing areas is difficult due to poor infrastructure. In urban areas, fruits are mostly imported from neighbouring production areas in the countryside and purchased from open markets during seasonal abundance. Thus, consumption in these areas depends to some extent on the purchasing power.

Previous research had reported vitamin A deficiency in Cameroon as public health problem (Gouado et al., 1998, 2005; Kollo et al., 2001). This problem is due to low accessibility, affordability and low intake of fruits or vegetables which is often associated to cultural beliefs and lack of knowledge on their importance.

Among the great varieties of fruits found in Cameroon, Mangifera indica L. and Carica papaya of different cultivars are among the most important in terms of production and marketing. Because they are succulent, exotic and delicious in taste and flavour, mango and papaya are popular. Their high carotenoid content could provide a high vitamin A value and anti-oxidative capacity to people living in vitamin A-deficient areas. But fresh mango and papaya have a short shelf life. In Cameroon and most tropical areas, these fruits are principally consumed in the form of fresh slices. Nowadays, small-scale industries have started to produce mango and papaya juice. As solar energy is free in these areas, if these fruits are dried and stored properly, perhaps, they could be all year round source of provitamin A. Based on the form in which it is consumed, the bioavailability of the main carotenoids in mango and papaya is likely to vary.

In this study, we determined the systemic levels of carotenoids provitamin A, lycopene and lutein in mango and papaya consumed in three different forms: dried, fresh and juice.


Study area

This study was conducted in four different sites.

  • The samples (fruits, subjects, administration of test meals and blood collection) were prepared at the University of Douala, Cameroon.

  • Analysis of carotenoids in fruits and serum retinol samples by high-performance liquid chromatography (HPLC) was carried out at the University of Ouagadougou, Burkina faso.

  • Isolation of serum chylomicrons at Max Planck Institute of biophysical chemistry, Göttingen, Germany.

  • Analyses of carotenoids (α-carotene, β-carotene, lycopene, β-cryptoxanthin, zeaxanthin, luteine) and vitamins (α-tocopherol and retinol) in chylomicrons were performed at the Department of Physiology and Pathology, Institute of Nutritional Science, University of Potsdam, Germany.

Preparation of subjects

After information campaign, through posters and oral announcements (in the Faculty of Science, University of Douala) on the objectives of the study and operations to be carried out, selection of subjects was done on those who voluntarily accepted to participate in the project. These volunteers were students at the Faculty of Science, University of Douala. They were eligible if they met the following criteria: during the period of the study, they were not using oral contraceptive, not consuming alcohol, cigarettes or any medication. They were not to have any gastrointestinal disease such as ulcer, not using drugs or food supplements a month before the study period and aged between 22 and 27 years. Following these conditions, 14 volunteers were retained for the study (7 for the mango test and 7 for papaya test). They gave their informed consent before participation. The protocol of the study was approved by the ethical committee for clinical studies at the University of Douala.

After three information seminars with these subjects, the program for interventions was elaborated jointly with the subjects. For each intervention, the subjects were to stay on diets poor in vitamin and provitamin A. To achieve this, the subjects were to keep a list of foods that they were not allowed to eat during this period. In the same week they were de-parasitized (500 mg of Mebendazole for intestinal parasites and nine tablets of Amodiaquine: 3 tablets per day during 3 days to prevent malaria infection). On the intervention day, the fasting subjects were received early in the laboratory. Blood collection (T0) was done with the aid of a catheter placed at the level of the elbow. Parameters such as weights and heights were determined before the first blood collection. The test meal made of a tin of yoghurt (130 g) as source of fat, a slice of bread (63 g) and mango or papaya fruit, containing various carotenoids (Table 3), between 100 and 568 g, depending on the mode of consumption (Table 2) then followed. Two other blood collections were made at 4 (T4) and 8 (T8) hours respectively after the test meal.

Preparation of fruit

Fruits were collected in Mungo region of Cameroon, and transported in ice baths to the laboratory. They were selected according to the maturation stage; the fruits were washed under tap water and peeled. The edible portions were sliced, weighed and processed in the following forms:

Juice: Fruits were homogenized using a blinder ‘moulinette’, water was added to facilitate homogenization and to reduce the viscosity of the product. The juices produced, without additive, were weighed and served to the subjects.

Fresh slices: Fruits were cut into slices, weighed and served.

Dried slices: Slices of fresh fruit were dried using a gas drier (the procedure started at 80°C and ended at 40°C) until residual moisture content was 12%.

For each form of fruit, the moisture content was determined (AOAC, 1980), and then the portion of samples to be used for carotenoids analysis was kept frozen in plastic cryotubes. The calendar intervention of different treatments is summarized in Table 1.

Table 1 Intervention program for the different treatments

Each treatment lasted for a day. The different experimental phases consisted of 2 weeks of acclimatization, during which the subjects were on their normal food habits; in the third week, their diets were deprived of vitamin A and provitamin A before they were subjected to the test meal.

Serum preparation

The serum obtained by centrifugation (3000 r.p.m.. for 5 min) of venous blood was collected and stored in cryotubes at −20°C for subsequent analysis. All these operations were done in the dark to avoid photodegradation of the vitamin and carotenoids. Samples that were haemolysed were discarded.

Triglycerides in the sera were analysed by colorimetric method, using the enzyme kits (Thermo trace; Australia).

Analysis of vitamin A in serum

Serum samples were unfrozen and left to attain ambient temperature. Then, 500 μl of absolute ethanol containing retinyl acetate as internal standard was used for precipitating protein in 200 μl of serum; 1 ml of n-hexane was used twice for vitamin extraction. After centrifugation at 3000 r.p.m. for 5 min at −5°C, the n-hexane phase was removed and introduced into another tube, which was then evaporated under nitrogen. The residue thus obtained was collected into acetonitrile and passed through Sonicator (Bioblock Scientifique 88169) at 10°C for 30 min. Sixty microlitres of the extract was then injected into the loop of the HPLC system. The elution was done under the following condition:

flow speed of the mobile phase (methanol acetonitrile water: 93:5:2): 2 ml min−1; average pressure: 1650 (1000–3500); wavelength of detection: 325 nm; range: 0.01AUFS; rise time: 0.3 s. The characteristics of the equipment were as follows, a supelcosil column LC-18, length, 25 cm, diameter, 4.6 mm, size of particle, 5 μm; a pre column; a pomp, Alltech 426 type, an integrator HP 3395; a lamp (Diode Array detector) Linear UVIS 200.

Analysis of carotenoid contents of the fruits

The fruits samples were unfrozen and left to attain ambient temperature. The extraction of carotenoids was done in a mixture of ethanol-hexane containing 0.01% BHT as antioxidant. After centrifugation at 3000 r.p.m. for 5 min at −5°C, the hexane phase was removed and introduced into another tube. This procedure was repeated until there was complete decoloration of the residue. The hexane phase obtained was evaporated under nitrogen. The residue was then collected into acetonitrile and passed through Sonicator (Bioblock Scientifique 88169) at 10°C for 30 min. Sixty microlitres of the extract was injected into the HPLC system. The elution was performed under the same conditions as in the case of the serum analysis, except that the wavelength of carotenoids detection was 450 nm (Taungbodhitham et al., 1999).

Isolation of chylomicrons from the serum

Chylomicrons were isolated using the method described by Van Vliet et al. (1995). Briefly, serum was unfrozen by a simple agitation in a water bath at 10°C. Serum (1 ml) was transferred into a polyallomer tube of 4 ml and a recovery of 3.4 ml of NaCl (d=1.006 g l−1—Merck, KgaA 64271 Darmstast, Germany), ultracentrifuged with a Swing-out type TFT 41.14 rotor (Kontron instruments) at 130000 g min−1 at a temperature of 10°C, for 30 min using Beckman L8-70 M (7A7221 series). Supernatant 0.8 ml containing chylomicrons was then removed and stored in test tubes at −40°C until used for carotenoids analysis.

Analysis of carotenoids in chylomicrons

Over a layer of 200 μl of chylomicron, a total of 200 μl of ethanol containing 0.05% BHT was added in a sealed test tube and homogenized using a vortex (MS2 minishaker, IKA, Janke and Kunkel, Staufen, Germany). Then 1 ml of hexane was added, agitated at 10 r.p.m. for 15 min using an agitator (Fröbel labor technik GmBH, Lindau, Type PR 50 L Germany). After centrifugation at 3800 r.p.m. for 10 min, the hexane layer was transferred to another labelled tube . This procedure was repeated twice. The hexane phase collected was then evaporated under nitrogen at 40°C. The residue was collected into 100 μl of isopropanol, mixed using a vortex mixer for 2 min and sonicated for 5 min. The supernatant obtained was transferred into the special tubes, and on HPLC Model Waters 515. (Schweigert et al., 2003).

Statistical analysis

ANOVA was used to compare means, Student's t-test and Mann–Whitney test was used respectively for comparison of two means, paired and unpaired. (Schwartz, 1991).


For the different cases, the composition of the test meals (Table 2 and Table3) was determined. The optimum required for maximum absorption of carotenoids, is 6 g of lipids. The retinol activity equivalent used was from IOM (2001).

Table 2 Test meal composition
Table 3 Quantity of carotenoids received per subject

Table 4 presents the different characteristics of the subjects. The average was 24 and 25 years and about 21.55 and 22.75 kg m−2 for body mass index, respectively, for subject fed mango and papaya. Serum triglyceride levels varied with the different groups. β-Carotene was the most abundant carotenoids in the different forms of mangoes, while in papaya, lycopene and β-carotene were found to be highest (Table 3).

Table 4 Characteristics of the subjects

The different carotenoids found in the fruits were also found in the chylomicrons with levels varying depending on the mode of consumption. From the results of the effect of consumption of the different fruit forms on the serum retinol and α-tocopherol (Figures 1a and b) levels, for each of the forms consumed, variation between T0 and T8 were not significant. In effect, tests carried out between mango juice and fresh mangoes at T0 and at T8 did not show any significant difference (P=0.275, and P>0.05). On the contrary the same tests done between mango juice and dried mangoes, and between fresh and dried mangoes showed significant differences at T0 (P=0.016 and P=0.001) and at T8 (P<0.05). The same situation also occurred in papaya (Table 5a and Table 5b), indicating that fresh mangoes and papaya had the highest systemic levels of carotenoids provitamin A. When comparing the mode of consumption at the start T0 and end of the experiment T8, using Fisher's test, significant variations were observed (T0: F(2,21)=11.299; P<0.001 and T8: F(2,21)=14.256; P<0.001). Comparing the mode of consumption of only two parameters in each case, significant variations were observed between the juice and dried fruits, and between fresh and dried fruits (P<0.001).

Figure 1

Mean retinol and α-tocopherol levels in the chylomicrons of subjects fed (a) mangoes, (b) papaya. a-Toco, α-tocopherol; Ret., retinol; T0, fasting (control); T4, 4 h after the test meal; T8, 8 h after the test meal.

Table 5a Mean values of serum retinol after consumption the different forms of mangoes
Table 5b Mean values of serum retinol after consumption of the different forms of papaya (T0 and T8)

The three forms of consumption therefore revealed three different absorption models (Figures 2a and b). For dried mangoes, α- and β-carotene did not attain the stationary phase within the first 8 h of the experiment. In the case of mango juice, lutein and β-carotene attained the stationary phase within 8 h after consumption of the test meal. For papaya, α- and β-carotene also attained the stationary phase at the end of 8 h of the experiment. This indicates that the quantities absorbed are at an optimum or the bioavailability is best at this period. This is confirmed in Figure 3 where the average levels of triglycerides in the chylomicrons and the three different models of each fruit are presented. Significantly positive correlation were also found between major carotenoids found in chylomicrons (lutein, α-and β-carotene) at different periods (T0, T4 T8). However for each carotenoid, the variations of average levels with the different times were not significant.

Figure 2

Mean carotenoids levels in the chylomicrons of subjects fed (a) mangoes, (b) papaya. Lut., luteine; Zea, zeaxanthin; b-cry, β-cryptoxanthin; a-caro, α-carotene, b-caro, β-carotene; Lyc., lycopene; T0,control; T4, 4 h after the test meal; T8,8 h after the test meal.

Figure 3

Mean levels of triglycerides in the chylomicrons of subjects fed mangoes and papaya. T0, control; T4, 4 h after the test meal; T8, 8 h after the test meal.


From the results obtained, it is shown that consumption of mangoes or papaya in any of the forms indicated (fresh slices, dried or juice) contribute enormously to lutein, α- and β-carotene in the chylomicrons of the subjects. There was also a slight increase in serum retinol after 8 h of eating the test diet. In several studies on the bioavailability of carotenoids, the subjects were supplemented for several weeks (Micozzi et al., 1992; Ribaya-Mercado et al., 1995). This therefore allows for proper appreciation of the serum response. But since 1995, after the studies of Van Vliet et al. (1995), bioavailability studies by postprandial response as used in the present study are increasingly being used because they are less tedious and fastidious for the subjects.

Contribution of cryptoxanthin and zeaxanthin were slow compared to the other carotenoids. The absorption of these carotenoids was influenced by many factors, as initially described by Van Het Hof et al. (2000). Bioavailability, which is the fraction of the nutrient ingested and available in the organism for its physiological functions and for storage (Jackson, 1997), is modulated by a number of metabolic steps that control the liberation of nutrients from the foods to the intestinal mucous (bioaccessibility), their intestinal absorption and transport to the different sites. In the case of carotenoids, they are absorbed at the level of the mucous of the duodenum by the mechanism of passive diffusion. The principal factor limiting the bioavailability of carotenoids in foods is its liberation from the food matrices where they are enclosed and dissolved in lipid droplets (Parker, 1996). In fruits, carotenoids are found in the chromoplast (Erdman et al., 1993), hence limits their liberation during digestion. It is thus better to transform the fruits (juice or fresh slices) before consumption. This thus permits breaking of the cell walls and dissociation of protein–carotenoid complex. Molecular simplification by grinding or homogenization reduces the effect of the molecular matrices to pass through. The sizes of the particles are reduced thereby facilitating digestion as opposed to fresh or dry slices that needs to be mashed. This is the reason why saturation of the cells occurred after 4 h of ingestion of papaya and mangoes in juice form.

The heat produced in the course of grinding is also likely to affect the bioavailability of carotenoids positively. The heat increases the dissociation of the protein–carotenoid complex or causes dispersion of the carotenoid crystals. This was observed in some studies on lycopene in products of tomato (Stahl and Sies, 1992). These results are in concordance with this hypothesis because differences were observed, on the one hand, between mango juice and dried mangoes (P=0.16) and on the other between fresh slices and dried mangoes (P=0.001). Same results were observed for papaya. It is evident that drying causes the liberation of carotenoids in the cellular matrix more difficult because the cell walls become harder, thus making it more difficult to digest as compared to fresh slices or juice. One of the direct consequences of drying could be the slowing the process of absorption. Thus in the case of dried mangoes, 8 h after administration of the test meal, saturation was not observed. This is a sign of slow digestion due to dehydration, which makes the liberation of nutrients more difficult. This phenomenon is not seen in papaya, where fibre was less as compared to mangoes. Rock (1997) and Williams et al. (1998) showed that the amount of fibre in foods and their exact nature are major factors that can inhibit the liberation of carotenoids from the matrices. In effect, fibre traps the carotenoids and acts on the bile acids favouring the faecal elimination of fat and liposoluble components. Drying helps dehydrate the fibres making it harder. However, in dried mangoes and papaya, the advantage is with their low water levels that favour storage for several months. In fresh mangoes, 4 h after eating the test meal, there is a rise in the level of α- and β-carotene in the chylomicrons. This can be attributed to the effect of other nutrients or enzymes that were not destroyed by drying and grinding.

Mango and papaya have a short shelf life, thus drying or processing these fruits increased their availability. But during processing and drying, carotenoids are susceptible to isomerization and oxidation, due to the presence of oxygen, exposure to light, heat treatment or destruction of the food matrix that protects the carotenoids. The consequence being the loss or reduction of biological activity.

Saturation of lutein and β-carotene was observed at T4, in signifying that they were absorbed best in that form. In effect, lutein being the most polar can easily be incorporated into the micelles.

The quantity and quality of food lipids constitute a primordial factor affecting bioavailability of carotenoids (Prince and Frisoli, 1993). Transportation of carotenoids and their products of hydrolysis from the intestines to the blood vessels and organs like liver, tissues is assured by lipoproteins rich in triglycerides (Parker, 1996). Absorption of carotenoids at the level of the cells of the mucus of the duodenum is best when lipids are present because of their lipophilic nature. From several reports, it is found that 3–5 g of lipids is the optimum level of lipids required for proper absorption of its carotenoids (Jalal et al., 1998; Roodenburg et al., 2000; Van Het Hof et al., 2000). In the present study, there was more than 6 g of lipids in the test meal indicating that this factor could not have affected the bioavailability of these nutrients, and thus not a source of variability.

Interaction between many nutrients in a food material or meal is also an important factor that affects bioavailability of carotenoids. Because carotenoids use the same route of absorption, it is possible to have positive or negative interaction among them. Hence, we had a significantly positive correlation between luteine, α- and β-carotene, a phenomenon that has been shown by many other authors (Böhm and Bitsch, 1999; Van Den Berg, 1999). Competition between β-carotene and lycopene or vitamin E has been shown in the works of Fuhrman et al. (1997).

Lycopene, abundant in papaya, administered to subjects were detected in very low levels in the chylomicrons. Stahl and Sies (1992) suggested that the cis form of lycopene is preferentially absorbed in the trans form. Thus the cis form found in processed tomatoes had a better bioavailability than the trans form because they were very short and more soluble in bile salts, and thus are better incorporated into the chylomicrons (Erdman, 2005). It could have been chemically degraded by the acid of the stomach or modified enzymatically at the level of mucus cells (Boileau et al., 2002).

These results show that fresh slices of mangoes and papaya had the best bioavailability values, followed by the juice form. Drying and processing of these fruits though geared at increasing their availability even at off seasons, causes slight reduction in their vitamin A activity. However, regular consumption of the fruits in these forms will in the long run ameliorate the vitamin A status of the population.


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We thank professors W Engel and Ahmed Mansouri from Max Planck Institute of Biophysics and Chemist, Göttingen, Germany, for the isolation of chylomicron in their laboratory. We gratefully acknowledge the Technical assistance of Hurtienne A. This study was funded by The International Foundation for Science (IFS) and Nutrition Third world (NTW) (Grant E -3584-1 to IG). The UNESCO fellowship allowed the first author to travel to Germany for HPLC analyses

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Gouado, I., Schweigert, F., Ejeh, R. et al. Systemic levels of carotenoids from mangoes and papaya consumed in three forms (juice, fresh and dry slice). Eur J Clin Nutr 61, 1180–1188 (2007).

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  • papaya
  • mangoes
  • carotenoids
  • chylomicrons

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