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Relevant physicochemical properties and metabolites with functional


properties of two commercial varieties of Peruvian Pouteria lucuma

Article  in  Journal of Food Processing and Preservation · April 2020


DOI: 10.1111/jfpp.14479

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Received: 6 August 2019    Revised: 27 October 2019    Accepted: 26 March 2020

DOI: 10.1111/jfpp.14479

ORIGINAL ARTICLE

Relevant physicochemical properties and metabolites with


functional properties of two commercial varieties of Peruvian
Pouteria lucuma

Diego García-Ríos1  | Ana Aguilar-Galvez1 | Rosana Chirinos1 | Romina Pedreschi2 |


David Campos1

1
Universidad Nacional Agraria La Molina,
Instituto de Biotecnología, Lima, Peru Abstract
2
Pontificia Universidad Católica de Two commercial varieties of Peruvian Pouteria lucuma fruits namely “Seda” and
Valparaíso, Escuela de Agronomía,
“Beltrán” were characterized in terms of their physicochemical properties as well as
Valparaiso, Chile
their primary and secondary metabolites content and profile. Free sugars, dietary
Correspondence
fiber, and starch comprise the main components in both varieties. Phenolic com-
David Campos, Instituto de Biotecnología,
Universidad Nacional Agraria La Molina, Av. pounds derived from flavanoids (flavan-3-ols), gallic acid, and their derivatives were
La Molina s/n, Lima, Valparaiso, Peru.
identified. Xanthophylls were tentatively identified based on their UV-vis spectra and
Email: dcampos@lamolina.edu.pe
enclosed the majority of the carotenoids found in lucuma. Additionally, both varieties
Funding information
showed to be sources of lipophilic compounds such as tocopherols (α, β, and γ) and
Vicerrectorado de Investigación—
Universidad Nacional Agraria La Molina; triterpenoids. The triterpenoid α-amyrin was identified in a Pouteria fruit for the first
Fondo Nacional de Desarrollo Científico,
time. The in vitro antioxidant capacity (AoxC) of the lipophilic fraction represented
Tecnológico y de Innovación Tecnológica-
FONDECYT, Grant/Award Number: approximately 30% of the total AoxC. These results show that both lucuma varieties
124-2015-FONDECYT
are rich sources of compounds with technological and functional properties with po-
tential application in the food industry.
Practical applications
Lucuma is a characteristic Peruvian fruit. To date, it is mainly used by the regional ice
cream industry and it has been included recently in some confectionery products.
Due to its high dietary fiber, carotenoids and sugar content it could be used as an
alternative to the use of refined sugars and artificial colorants in the dairy and bakery
industry.

1 |  I NTRO D U C TI O N Other Pouteria species such as P. campechiana (canistel) (Costa,


Wondracek, Lopes, Vieira, & Ferreira, 2010), P. viridis (Ma et al., 2004),
The genus Pouteria belongs to the Sapotaceae family and it is dis- and P. macrophylla (da Silva, Gordon, Jungfer, Marx, & Main, 2012)
tributed in the tropical and subtropical regions of Asia and America. have also been reported as rich sources of bioactive compounds.
Many species are of commercial importance since they are used as Lucuma (P. lucuma) is a fruit native to the Andean region, found
food and in traditional medicine (Silva, Simeoni, & Silveira, 2009). in Peru and Chile; it can be cultivated from the sea level to an alti-
One of the most studied species of this family is P. sapota (mamey) tude of 3,000 m.a.s.l (Jordan, 1996). It presents an ovoid or elliptical
which has been shown to be a source of many bioactive compounds shape depending on the cultivar, a diameter variable between 7.5
such as polyphenols, carotenoids, and tocopherols (Ma, Yang, Basile, and 10 cm, thin green or yellow-green skin and sweet yellow-or-
& Kennelly, 2004; Yahia, Gutiérrez-Orozco, & Arvizu-de Leon, 2011). ange flesh (Yahia & Gutiérrez-Orozco, 2011). In Peru, there are two

J Food Process Preserv. 2020;00:e14479. wileyonlinelibrary.com/journal/jfpp |


© 2020 Wiley Periodicals LLC     1 of 11
https://doi.org/10.1111/jfpp.14479
|
2 of 11       GARCÍA-RÍOS et al.

types of lucuma: "Seda" and "Palo." The "lucuma Seda" has a pulp (Carbohydrates Kit, CAR10), organic acid standards: L-ascorbic,
with a smooth texture when ripe, floury, intense yellow color, soft citric, malic, quinic, succinic and tartaric were purchased from
on the palate and sweet and it is suitable for consumption as fruit. Supelco (Kit 47264) and standards of methyl esters of fatty
The "lucuma Palo," in contrast, when ripe has a pulp of hard texture, acids (FAME Mix, 37 components) were purchased from Restek.
not suitable for fresh consumption, which makes it more appropri- Acetone, methylene chloride, hexane, ethanol, anhydrous sodium
ate for industrial processing (Sistema Integrado de Información de sulfate and HPLC grade solvents (acetonitrile, methanol, hexane)
Comercio Exterior, 2015). However, color, aroma and other thermo- were purchased from J.T. Baker, MS-grade methanol LiChrosolv®
sensitive compounds could be affected in different degrees during from Merck. Sodium carbonate, sodium chloride, 2-propanol, and
postharvest storage and/or processing conditions as consequence Folin–Ciocalteu reagent were purchased from Merck. Glacial acetic
of changes and interactions among the compounds present. Thus, it acid was purchased from Fermont and potassium hydroxide was
is important to be aware of the components present to prevent po- purchased from Mallinckrodt.
tential changes in the physicochemical, organoleptic and bioactivity
and control these changes during processing or storage.
Recently, lucuma has attracted the attention of consumers and 2.2 | Characterization and quantitative analysis
researchers because it constitutes a source of various compounds of
interest for their antioxidant properties such as carotenoids and phe- 2.2.1 | Physico-chemical characteristics
nolic compounds (Dini, 2011; Erazo, Escobar, Olaeta, & Undurraga,
1999; Fuentealba et al., 2016). To date, the characterization and Moisture, ash, lipid, and protein (N × 6.25) were determined ac-
quantification of the aforementioned metabolites in lucuma have cording to the AOAC (2007) methods. Moisture was determined in
not been fully completed. Also, no previous study has provided in- both fresh and lyophilized pulp. Total dietary fiber and starch were
formation about the content of tocopherols or terpenoids in lucuma. determined according to the AOAC method (AOAC, 2007), results
Therefore, the objective of this study is to identify and quantify the were expressed as g/100 g of DW. Soluble solids, pH, and titrat-
physicochemical characteristics as well as compounds with bioactive able acidity were performed following the methods described in the
properties of lucuma fruits of two commercial varieties: "Seda" and AOAC (2007). Reducing sugars were determined as recommended
"Beltrán". by Miller (1959) with some modifications and were expressed as glu-
cose. Color determination (L*, a*, and b* values) was carried out both
in the peel and in the pulp as the average of six measurements at the
2 |  M ATE R I A L S A N D M E TH O DS equatorial region using the Konica Minolta Chromameter colorim-
eter (CR-400, Konica Minolta).
2.1 | Materials and reagents

2.1.1 | Fruit material 2.2.2 | Determination of sugars, sugar-alcohols


and organic acids
Two varieties commercially known as "Seda" and "Beltrán" from the
province of Huaral, Peru and purchased directly from a local pro- Sugars, sugar-alcohols and organic acids were extracted accord-
ducer were used. The fruits were harvested at commercial maturity ing to Pérez, Olías, Espada, Olías, and Sanz (1997) with certain
(yellow peel coloration under the calix), transported to the labora- modifications. Briefly, 0.5 g of lyophilized lucuma was extracted
tory and stored for 5 days at room temperature, until they reached twice with 25 ml of 95% ethanol for 10 min for each extraction.
edible ripeness. Subsequently, they were peeled, lyophilized, and The extracts were mixed and concentrated in a rotary evapora-
stored at −40°C until analysis. tor at 50°C, until dryness. The residue was dissolved with 4 ml of
0.2 N H2 SO 4 containing 0.05% EDTA. Eight hundred microliter of
the solution was taken and injected into a Sep-Pak C18 cartridge
2.1.2 | Chemicals (WAT036905, Waters, Ireland), then eluted with 10 ml of 0.2 N
H2 SO 4. The content of sugars and sugar-alcohols was determined
Butylated hydroxytoluene (BHT), β-carotene, myo-inositol, phe- following the method described by Campos, Aguilar-Galvez, and
nolic acid standards (ellagic and gallic), flavanones (hesperetin), to- Pedreschi (2016); while organic acids according to the protocol
copherol standards (α, β, γ, and δ-tocopherol), α-amyrin, β-sitosterol, described by Aguilar-Galvez, Guillermo, Dubois-Dauphin, and
and cycloartenol were purchased from Sigma Chemicals Co. (St. Campos (2011). In both cases, a Waters 2695 Separation Module
Louis). Flavan-3-ols (catechin, epicatechin, gallocatechin, and epi- (Waters) equipped with an autoinjector, a 2414 refractive index
gallocatechin gallate) and procyanidins (procyanidin B1 and B2) and detector 996 photodiode array detector, respectively, and
were purchased from ChromaDex ™ (Santa Clara). Sugar stand- the Empower software (Waters) were used. The results were ex-
ards: fructose, glucose and sucrose were purchased from Sigma pressed in mg/ g of DW.
GARCÍA-RÍOS et al. |
      3 of 11

2.2.3 | Determination of L-ascorbic acid dryness with nitrogen, then re-suspended in 4 ml of methanol con-
taining 10% potassium hydroxide (w/v). The solution was allowed
Sample treatment for the quantification of L-ascorbic acid was per- to stand for 16 hr in the dark at 4°C in a nitrogen-saturated atmo-
formed as reported by Sánchez-Moreno, Plaza, de Ancos, and Cano sphere and 4 ml of hexane and 4 ml of saturated NaCl solution were
(2003) with some modifications. One gram of lyophilized sample was added. The organic phase was recovered and the aqueous phase
homogenized with 10 ml of 3% metaphosphoric acid solution and 8% re-extracted with hexane. Both organic phases were mixed, brought
acetic acid for 10 min. The mixture was centrifuged at 2,795 g at 4°C to dryness with N2 and re-dissolved in 1 ml of methylene chloride:
for 25 min (Hettich, model MIKRO 220R). The extract was analyzed ethanol (65:35, v/v).
by HPLC-PAD. A Prodigy ODS3 100A (5μm, 250 × 4.6 mm ID) col- The analysis was performed by HPLC-PDA according to the
umn (Phenomenex) was used. The mobile phase was composed of method of Kao, Loh, Inbaraj, and Chen (2012) with slight modifica-
25 mmol/l KH2PO4, pH 2.5 at a flow rate of 1 ml/ min. Samples were tions. Spectral data were recorded from 330 to 550 nm during the
filtered prior to HPLC injection. Ten microliter of sample was injected whole run. A YMC TM carotenoid (5 μm, 250 × 4.6 mm ID) column
and run for 15 min at 40°C. L-ascorbic acid was detected and quanti- (Waters, Ireland) was used for separation at 30°C. The mobile phase
fied at 245 nm. The results were expressed in mg/ 100 g of DW). was composed of solvent (A) methanol: acetonitrile: water (79:14:
7, v/v/v) and solvent (B) methylene chloride. The solvent gradient
was as follows: 5% B for 9 min, 5%–15% B in 14 min, 15%–17% B in
2.2.4 | Determination of fatty acids 10 min, 17%–29% B in 2 min, 29%–30% B in 10 min, 30%–34% B in
21 min and returned to 5% B in 1 min and kept at those conditions for
The lipophilic fraction was extracted from 100 mg of lyophilized 5 min. A flow rate of 1.0 ml/ min was used and 20 μl of sample was
lucuma and 500 μl of 96% methanol, 750 μl of Milli-Q water, and injected. Carotenoids were identified by comparison of retention
375 μl of chloroform were added. The samples were sonicated for times and absorption spectra with data reported in the literature.
30 min and then centrifuged for 10 min at a maximum speed. The
chloroform phase was recovered and the methanol/ water phase
was re-extracted with 375 μl of chloroform. The chloroform phases 2.2.6 | Total phenolic compounds and profile
were mixed and evaporated in a nitrogen-saturated atmosphere. The
fatty acid profile was determined according to the method proposed The extraction of phenolic compounds was carried out according
by Meurens, Baeten, Yan, Mignolet, and Larondelle (2005) with slight to the method proposed by Ma et al. (2004) with slight modifica-
modifications. The fatty acids present in the residue were converted tions. Half a gram of lyophilized lucuma was mixed with 25 ml of 80%
to methyl esters and one microliter was injected in a gas chroma- acetone and stirred for 90 min. Subsequently, the extract was cen-
tograph GC-2010 plus Shimadzu equipped with an FID-2010 flame trifuged at 2,800 g and stored in dark and nitrogen-saturated atmos-
ionization detector and AOC-20i autoinjector. The column used was phere at −20°C until analysis. Total phenolic compounds (TPC) were
a Restek Rt-2560 (0.2 μm, 100 × 0.25 mm ID). The methyl esters of determined according to the method of Singleton and Rossi (1965)
the fatty acids were identified and quantified by comparing reten- by colorimetric reaction with Folin–Ciocalteu reagent. The content
tion times with known and previously injected standards. The results of TPC was expressed in mg of Gallic equivalent (AGE)/ g DW.
were expressed as a percentage relative to the total of fatty acids. Phenolic profile was assessed following the method of Chirinos
et al. (2008) with slight modifications. Phenolic extracts were sep-
arated on HPLC-PDA. Spectral data were recorded from 200 to
2.2.5 | Total carotenoids and profile 700 nm during the whole run. An X-terra RP18 (3.5 µm, 250 × 4.6 mm
ID) column (Waters) was used for separation at 30°C. The mobile
Carotenoid extraction was carried out according to the method re- phase was composed of solvent (A) water: formic acid (95:5, v/v) and
ported by Andre et al. (2007) with slight modifications. Lyophilized solvent (B) acetonitrile. The solvent gradient was as follows: 0%–15%
lucuma pulp (0.5 g) was mixed with 5 ml of acetone: hexane (1:1, v/ v), B in 40 min, 15%–45% B in 45 min, and 45%–100% B in 10 min. A
homogenized and stirred in an ice bath for 30 min. The extract was flow rate of 0.5 ml/min was used and 20 µl of sample was injected.
centrifuged at 2,795 g for 15 min at 4°C, and the supernatant was Samples were filtered prior to HPLC injection. Phenolic compounds
collected. The extraction was repeated in the cake twice for 10 min were identified by comparing their retention time and UV-vis spec-
each, maintaining the initial extraction conditions. The supernatants tral data to known previously injected standards.
were combined and evaporated to dryness in a rotary evapora-
tor at 40°C, and finally suspended in 2  ml of acetone. Total carot-
enoids were determined by spectrophotometry (Genesys™ 20 Vis 2.2.7 | Determination of tocopherols
Spectrophotometer, Thermo Scientific) at 450 nm and the content
was expressed in mg of β-carotene equivalents/ 100 g DW. Tocopherols were quantified and identified following the method-
To determine the profile of carotenoids, saponification of the ology proposed by Amaral, Rui, Seabra, and Oliveira (2005). Half
extract was carried out, an aliquot of the extract was evaporated to a gram of lyophilized lucuma pulp was mixed with 100 μl of BHT
|
4 of 11       GARCÍA-RÍOS et al.

solution (10 mg in 1 ml of hexane), 2 ml of ethanol, 4 ml of hexane 2.2.9 | In vitro hydrophilic and lipophilic
and 2 ml of saturated NaCl solution. The mixture was homogenized antioxidant capacity
for 1 min in a vortex and then centrifuged for 4 min at 2,505 g at 4°C.
The upper phase was collected and the sample was re-extracted Antioxidant capacity (AoxC) was reported by the TEAC (Trolox equiva-
twice with 2 ml of hexane. The extracts were combined and taken to lent antioxidant capacity) test recommended by Arnao, Cano, and
dryness with nitrogen and the residue was re-dissolved with 0.5 ml Acosta (2001) in the hydrophilic and lipophilic extracts, using the ABTS
of hexane. The extract was dried with anhydrous sodium sulfate reagent. The extracts were obtained according to the protocol of Wu
(0.5 g), centrifuged at 7,378 g for 2 min. et al. (2004) with some modifications. The lipophilic extract was ob-
Samples were separated using a HPLC equipped with a Waters tained from 1 gram of lyophilized lucuma pulp, homogenized in 10 ml
2475 multi fluorescence detector. An YMC-Pack SIL (3 µm, of hexane: dichloromethane (1:1, v: v) in an ice bath for 15 min and then
250 × 4.6 mm ID) column (YMC, Japan) was used for tocopherol centrifuged at 4,000 g for 10 min at 4°C. The extraction was repeated
separation at 35°C. The mobile phase was composed of n-hexane/ after recovering the supernatant under the same conditions as the first
2-propanol/acetic acid (1000/6/5, v/v/v). A flow rate of 1.4 ml/min extraction. The supernatants were mixed and evaporated on a rotary
under isocratic conditions was used. Ten microliter of sample was evaporator at 40°C under reduced pressure. Finally, the dry extract
injected. Samples were filtered prior to HPLC injection. The fluo- was re-suspended in 10 ml acetone and stored under a nitrogen at-
rescence detector was programmed at the excitation and emission mosphere at −20°C until analysis. The hydrophilic extract was obtained
wavelengths of 290 and 330 nm, respectively. Tocopherols were from the residue of the lipophilic extraction which was homogenized
identified and quantified by comparing their retention time to in 20 ml of acetone: water: acetic acid (70:29.5:0.5, v/v/v) at room
known previously injected standards. Results were expressed as temperature for 15 min. The extraction was repeated after the cen-
mg/100 g of dry matter. trifugation performed under the same conditions as for the hydrophilic
extract. The supernatants were mixed and stored under the same con-
ditions as for the lipophilic extract. The results of both methods were
2.2.8 | Phytosterols and triterpenoids expressed in μmol of Trolox equivalent (TE)/ g sample (dry basis).

The unsaponifiable extract was obtained according to the protocol


reported by Duchateau et al. (2002). Twelve and a half grams of ly- 2.3 | Statistical analysis
ophilized sample was mixed with 100 ml of hexane: acetone: etha-
nol (50:25:25, v/v/v) for 10 min under agitation and then 15 ml of Results represent the mean and standard deviation of three inde-
water was added and stirred for another 5 min. The lipid phase was pendent experiments (n = 3). T- tests were performed with a 95%
recovered and concentrated in a vacuum rotary evaporator at 60°C. confidence, using the Statgraphics Centurion XVI (StatPoint Inc.).
Hundred milligram of the lipid extract was saponified with 1 ml of
methanolic KOH at 70°C for 50 min, after the addition of the internal
standard (β-cholestanol). The unsaponifiable fraction was extracted 3 | R E S U LT S A N D D I S CU S S I O N
by liquid–liquid partition with 1 ml of distilled water and 5 ml of n-
heptane. The n-heptane extract was recovered and the extraction 3.1 | Physico-chemical characteristics
was repeated in the aqueous fraction twice. The n-heptane extracts
were mixed and dehydrated with anhydrous sodium sulfate. Trace The physico-chemical characteristics are shown in Table 1. The
1310 Gas chromatograph (Thermo Scientific, Rodano) was used, moisture contents were similar for both varieties (p > .05), 56.2 and
coupled to a triple Quadrupole TSQ 8000 Evo Mass Spectrometer 56.9% for Beltrán and Seda, respectively. The results are within
(Thermo Scientific, using a TG-5SILMS column (30 × 0.25 mm ID, the humidity range reported by Erazo et al. (1999) in six selections
0.25  μm film thickness) (Thermo Scientific). The chromatographic of Chilean lucuma (56.03 to 63.16%). The stage of maturity at the
separation was carried according to da Costa, Augusto, Teixeira- time of harvest and storage conditions can affect the moisture con-
Filho, and Teixeira (2010). The furnace temperature was programmed tent of the fruit (Alia-Tejacal et al., 2007; Fuentealba et al., 2016).
as follows: initially at 250°C (for 2 min), increased to 285°C at a rate of In general, the moisture content of lucuma is lower than that of
25°C/ min and an isothermal period of 32 min at 285°C. The injection other Sapotaceae such as mamey (75%) and sapodilla (82%) (Moo-
volume was 1 μl with a split ratio of 10. The temperature of the injec- Huchin et al., 2014), although a moisture value of 49.5% in canistel
tor was set at 300°C. Helium was used as a carrier gas. For the mass has been reported (Costa, Wondracek, et al., 2010). The total die-
spectrometric analysis, the ionization was performed by electronic tary fiber content was similar for both varieties (24.2%) and higher
impact (EI) in a positive mode at 70 eV and detection by scanning in than that reported in sapote (17.2%–21.5%) (Mahattanatawee et al.,
the range of 45 to 600 m/z was performed. Compound identification 2006; Moo-Huchin et al., 2014). For this reason, lucuma is an op-
was performed by comparing retention times and mass spectra with tion to meet the fiber requirements recommended for adults by the
previously injected standards and by comparison with the NIST 2.0 American Dietetic Association (between 20 and 35 g/day) (Marlett,
library. The results were expressed as μg/g of dry matter. McBurney, & Slavin, 2002).
GARCÍA-RÍOS et al. |
      5 of 11

TA B L E 1   Composition and physico-chemical characteristics for TA B L E 2   Content of sugars, myo-inositol and organic acids in
Beltrán and Seda lucuma varieties Beltrán and Seda lucuma varieties

Variety Variety
Components
Component Beltrán Seda (mg/g DW) Beltrán Seda
1 a a
Moisture   (g/100 g) 56.2 ± 3.7 56.9 ± 3.3 Sugars    
a b a
Protein (g/100 g DW) 4.3 ± 0.02 5.2 ± 0.05 Glucose 147.2 ± 18.5 140.9 ± 5.5a
a b a
Ash (g/100 g DW) 2.1 ± 0.08 2.51 ± 0.04 Fructose 155.9 ± 16.7 136.9 ± 10.6a
Lipids (g/100 g DW) 1.29 ± 0.06a 1.23 ± 0.03a Sucrose 57.6 ± 3.9a 48.0 ± 6.7a
a a
Total Dietary Fiber (g/100 g DW) 24.2 ± 1.4 24.2 ± 0.7 Sugar alcohols    
b
Soluble Fiber 4.5 ± 0.8 3.9 ± 0.5 myo-inositol 5.7 ± 0.4 9.9 ± 0.5a
Insoluble Fiber 19.7 ± 1.2 20.3 ± 0.5 Organic acids    
a b b
Starch (g/100 g DW) 15.6 ± 1.6 11.7 ± 0.5 Citric 1.7 ± 0.1 3.4 ± 0.4a
a a b
Reducing sugars (g glucose/100 g 27.2 ± 1.7 23.2 ± 3.5 Tartaric 0.55 ± 0.03 1.0 ± 0.2a
DW) Malic ND 1.6 ± 0.2
Soluble solids 21.7 ± 0.6b 23.4 ± 0.9a Quinic 14.3 ± 0.7a 14.9 ± 1.4a
pH 5.56 ± 0.06a 5.49 ± 0.04a Succinic 0.8 ± 0.01a 0.6 ± 0.1b
a
Titrable acid (% of citric acid) 0.28 ± 0.01 0.36 ± 0.02a L-ascorbic 0.68 ± 0.01 a
0.58 ± 0.02b
Peel Color    
Note: Different letters within the same row stand for significant
L* 50.7 ± 2.7a 46.2 ± 3.1a differences (p < .05).
a* 6.0 ± 2.5a −6.8 ± 2.0 b Abbreviation: ND, non detected.
a
b* 35.3 ± 5.2 22.3 ± 1.2b
Pulp Color     green - yellow for Seda. The appearance of yellow-orange tones is
L* 71.6 ± 2.3a 69.2 ± 3.6a attributed to the synthesis of carotenoids, as the fruit matures, chlo-
a* 16.6 ± 3.0 a
13.9 ± 5.4 a
rophyll degrades and carotenoids are synthesized (Eskin & Hoehn,
b* 68.8 ± 3.2a 52.6 ± 6.9a 2013; Li & Yuan, 2013). These differences in peel color might suggest
that the synthesis-degradation balance of chlorophyll during ripen-
Note: Different letters within the same row stand for significant
differences (p < .05). ing is different between both varieties of lucuma. In contrast, the
1
Moisture measured on the fresh pulp. color of the pulp showed no significant differences (p > .05) for any
of the parameters (L*, a*, b*). These values were higher than those re-
ported in mamey (Alia-Tejacal et al., 2007; Moo-Huchin et al., 2014).
The starch content was significantly higher in Beltrán variety,
but the content of reducing sugars was similar for both varieties.
Controlling the content of sugars and starch is important for the 3.2 | Determination of sugars, sugar-
conservation of the fruit because although it is possible to reduce alcohols and organic acids
the starch content (therefore, increase the content of sugars), this
increase could result in an over-ripe and easily affected fruit (Eskin & Results corresponding to primary metabolites (sugars and sugar al-
Hoehn, 2013). Seda presented an acidity (0.28%) lower than Beltrán cohols) are shown in Table 2. Most relevant sugars corresponded to
(0.36%); both values were similar to those reported for mamey (0.2 fructose and glucose. No significant differences in sugar contents
to 0.3%) (Alia-Tejacal et al., 2007). However, the acid values found were found (p > .05) for Beltrán and Seda, respectively with values of
were lower than most of those reported by Moo-Huchin et al. (2014) fructose (155.9 and 136.9 mg/g DW), glucose (147.2 and 140.9 mg/g
for tropical fruits, which have contents in the order of 0.3 to 1.9% DW), and sucrose (57.6 and 48.0 mg/g DW), respectively. The con-
acidity expressed as citric acid. No significant difference (p > .05) tents of fructose and sucrose were higher than those reported by
was found in the pH values for both varieties. The content of soluble Fuentealba et al. (2016) for lucuma biotype Leiva 1 (98.7 mg/ g and
solids was significantly higher (p < .05) in Seda (23.4%) than Beltrán 36.2 mg/g in DW, respectively); while the glucose content found in
(21.7%). These values were similar to those reported by Alia-Tejacal this work was lower than that reported by Fuentealba (170.9 mg/g
et al. (2007) for mamey (23%–27%). DW). The composition of sugars in the studied lucuma varieties is
Regarding color, differences were found in the peel of both vari- different from that reported in canistel where sucrose predomi-
eties, while the values of L* and b* were similar, differences for the nates (96.89 mg/g DW, Kubola, Siriamornpun, & Meso, 2011) and
value of a* were encountered with positive values (red) in Beltrán in mamey where non-reducing sugars, such as sucrose, represented
and negative values (green) in Seda. Very marked differences in the more than 70% of total sugars (38.96–69.35 mg/g DW, Alia-Tejacal
hue of the peel were found with yellow - orange for Beltrán and et al., 2007). Myo-inositol was the only sugar alcohol detected;
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the content in Seda was higher (9.9 mg/g DW or 4.3 mg/g FW) TA B L E 3   Content of saturated and unsaturated fatty acids,
than in Beltrán (5.7 mgg DW or 2.5 mg/g FW). These values were total carotenoids, total phenolic compounds, tocopherols and
antioxidant activity in two commercial lucuma varieties
greater than 2.1 mg/g DW and differed from the values reported by
Fuentealba et al. (2016) for lucuma biotype Leiva 1. Variety
Organic acid profile and content are shown in Table 2. The high-
Compounds Beltrán Seda
est content corresponds to quinic acid and similar contents of 14.3
Saturated fatty acids (% of total    
and 14.9 mg/g DW (p > .05) were found in both varieties. For other
fatty acids)
quantified organic acids, Seda variety had a significantly higher
Lauric (C12:0) traces 2.9
concentration of citric and tartaric acid, while Beltrán had a higher
Miristic (C14:0) 7.5 6.6
concentration of succinic and ascorbic acid, respectively. Malic
Pentadecanoic (C15:0) 3.6 5.4
acid was only detected and quantified in Seda variety. In other
Pouteria species such as mamey, malic acid was reported as the Palmitic (C16:0) 25 23.4

predominant organic acid (Alia-Tejacal et al., 2007). The greatest Stearic (C18:0) 20 17.7
difference observed between Seda and Beltrán corresponds to or- Unsaturated fatty acids (% of total    
ganic acids linked to respiration (citric, malic and succinic) and can fatty acids)

be attributed to the environmental conditions in which the fruits Palmitoleic (C16:1) 2.5 1.9

were stored after harvest, since the content of these acids tended Oleic (C18:1) 10 12.2
to decrease more rapidly at higher storage temperatures (Eskin & Linoleic (C18:2) 1.8 2.9
Hoenh, 2013). α-linolenic (C18:3) 29.6 27
ω6/ ω3 ratio 0.06 0.11
Total carotenoids (mg β-carotene 0.30 ± 0.01a 0.25 ± 0.02b
3.3 | Determination of L-ascorbic acid eq/ g DW)
Total phenolic compounds 2.50 ± 0.11a 2.38 ± 0.13a
The content of ascorbic acid was significantly different between (mg AGE/ g DW)
Seda (0.68 mg/g DW or 0.29 mg/g FW) and Beltrán (0.58 mg/g Tocopherols (μg/ g DW)    
DW or 0.25 mg/g FW) varieties, respectively. These contents α-Tocopherol 47.4 ± 1.6 b
59.4 ± 1.3a
were higher than that reported in lucuma Leiva 1 (0.19 mg/g DW) β-Tocopherol 0.68 ± 0.01b 0.75 ± 0.01a
by Fuentealba et al. (2016). Ascorbic acid contents in the range of γ-Tocopherol 7.1 ± 0.3 ND
0.2–1.2 mg/g DW have been reported in mamey (Mahattanatawee δ-Tocopherol ND ND
et al., 2006; Moo-Huchín et al., 2014). Pouteria species with the
Antioxidant capacity    
highest vitamin C content reported correspond to P. macrophylla (μmol TE/ g DW)
with 2.5 mg/g DW (Gordon, Jungfer, Alexandre, Guilherme, & Marx, Hydrophilic 19.3 ± 1.3a 17.3 ± 1.0 b
2011; da Silva et al., 2012) and canistel with 1.9 mg/g FW (Kubola
Lipophilic 8.7 ± 0.1a 7.4 ± 0.4b
et al., 2011).
Phytosterols and Triterpenoids    
(ug/g DW)
β-Sitosterol 4.44 5.27
3.4 | Determination of fatty acids
Cycloartenol 3.50 2.48
α-Amyrin 11.85 12.34
The contents of the different fatty acids are shown in Table 3. Both
lucuma varieties presented a similar profile of fatty acids. Within the Note: Different letters within the same row stand for significant
differences (p < .05).
saturated fatty acids, the most representative was palmitic acid that
Abbreviation: ND, non detected.
represents between 23% and 25% of total fatty acids. Among the
unsaturated fatty acids, α-linolenic acid (ω-3) was the most abundant
and constitutes between 27% and 30% of unsaturated fatty acids. It
is important to point out that this fatty acid is an essential nutrient 3.5 | Total carotenoids and profile
of the diet and plays an important role in human health due to its
association with the reduction of the risk of cardiovascular diseases, Total carotenoid content was significantly higher in the variety
cancer, osteoporosis, and immune disorders (Vilela et al., 2013). The Beltrán (0.3 mg of β - CE/ g DW) than Seda (0.25 mg of β - CE/g
content of linoleic acid (ω-6) was very low with respect to total fatty DW) (Table 3). These contents were similar to those reported by
acids, possibly due to its oxidation and subsequent cleavage by en- Fuentealba et al. (2016) for lucuma Leiva 1 and greater than those
zymes lipoxygenase and aldehyde lyase, which convert this acid into obtained by Erazo et al. (1999) who reported a range of 0.03–
volatile components such as aldehydes and ketones, which contrib- 0.05 mg of β - CE/g DW. In addition, the content of total carotenoids
ute to the aroma profile in the mature fruit (Eskin & Hoehn, 2013). is within the values normally reported for species belonging to genus
GARCÍA-RÍOS et al. |
      7 of 11

Pouteria such is mamey 1.44 mg of β - CE/g DW (Moo-Huchin et al., 2.8 mg of GAE/g DW (Mahattanatawee et al., 2006) were previously
2014). reported. Instead, in canistel a value of 5 mg of GAE/g DW (Kubola
HPLC-PDA analysis allowed the separation of 10 carotenoids et al., 2011) was reported and a much higher content (22.9 mg of
(Figure 1), eight of which were identified by comparing their reten- GAE/g DW) was reported in P. macrophylla (da Silva et al., 2012) and
tion times and absorption spectra to those reported by Kao et al. up to 29.2 mg of GAE/g DW according to Gordon et al. (2011). This
(2012). The main carotenoids found in lucuma pulp belong to the variation responds to multiple factors both environmental and spe-
family of xanthophylls. The epoxide forms such as neoxanthin and cific of the fruit, within the most important environmental factors
violaxanthin were more abundant in the Beltrán variety. In contrast, are the harvest season and location of the crop (Barceló, Nicolás,
the Seda variety presented hydroxylated carotenoids (lutein deriva- Sabater, & Sánchez, 2009) while factors inherent to the fruit cor-
tives) as the most abundant carotenoids. Traces of β-carotene were respond to the type of cultivar and maturity stage. Fuentealba et al.
detected in both varieties; however, the presence of derivatives such (2016) observed a drastic reduction in the content of TPC in lucuma:
as β-cryptoxanthin, zeaxanthin, and violaxanthin would indicate that at harvest maturity (131.6 mg of GAEg DW), the fruit naturally de-
the carotenoids in lucuma tend to accumulate in the form of xan- tached from the tree (45.3 mg of GAE/g DW) and then stored at
thophylls (possibly esterified) rapidly as maturation progresses (Li & 20°C for 1 week (0.7 mg of GAE/g DW). There can also be variations
Yuan, 2013). This process seems to begin even before reaching com- within the same cultivar or species (Barceló et al., 2009).
mercial maturity, as reported by Fuentealba et al. (2016). Regarding the nature of phenolic compounds present in lucuma
The nature of the carotenoids present in lucuma allows us to at edible ripeness, chromatographic analysis allowed the determi-
infer that these components could be significantly affected by a nation of eight main compounds in both varieties that showed a
conventional drying process extensively used for lucuma flour elab- similar phenolic compound profile (Figure 2). The majority of these
oration (Yahia & Gutiérrez-Orozco, 2011). Previous studies have re- compounds were tentatively identified and quantified as the deriv-
ported that hot air conventional drying significantly decreases the atives of gallocatechin, epigallocatechin, catechin and epicatechin
total carotenoid content in the chips of sweet potato (Bechoff et al., (peaks 1, 3, 4, 5, 6, and 9) based on their UV spectra. Other com-
2010). Most of the losses observed in sweet pepper during drying pounds identified were gallic acid (peak 2), ellagic acid (peak 8) and
corresponded to the xanthophils violaxanthin and zeaxanthin in ad- a derivative of hesperetin (peak 7). The nature of these compounds
dition to β-carotene (Topuz, Dincer, Sultan, Feng, & Kushad, 2011). was consistent with that reported by Fuentealba et al. (2016), who
identified gallic acid and a flavonoid derivative in the hydrolyzate
of lucuma phenolic compounds. Dini (2011) isolated and identified
3.6 | Total phenolics compounds and profile gallic acid and complex glycosides of kaempferol in lucuma flour.
The latter, however, was not identified in the present study. The
The TPC content in both varieties was similar, with 2.5 mg of occurrence of similarity between the absorption spectra and the
GAE/g DW and 2.4 mg of GAE/g DW for Beltrán and Seda, respec- difference between the retention times with respect to standards
tively (Table 3). These values were higher than those reported by could suggest that the lucuma phenolic compounds are probably
Fuentealba et al. (2016) for lucuma Leiva 1 (0.7 mg AGE/g DM). In glycosylated to different types of sugars or in different positions
mamey, values of 0.6 mg of GAE/g DW (Moo-Huchin et al., 2014) and (Gordon et al., 2011).

F I G U R E 1   HPLC-PDA carotenoid
profile obtained at 450 nm for the
varieties Beltrán (a) and Seda (b), tentative
identification and UV obtained data.
Identification was based on data reported
by Kao et al. (2012)*
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8 of 11       GARCÍA-RÍOS et al.

F I G U R E 2   HPLC-PDA profile for


phenolic compounds obtained at 280 nm
for the lucuma varieties Beltrán (a) and
Seda (b); identification and UV data for
the phenolic compounds

The compounds found in this study are similar to those previ- the Beltrán variety, could increase its stability even more before ox-
ously reported for other Pouteria species. In general, gallic acid, idation with respect to the variety Seda. These isomers according
catechins and gallocatechins are the most representative phenolic to Kamal-Eldin and Budilarto (2015) significantly contribute to the
compounds (Ma et al., 2004; da Silva et al., 2012). The authors point oxidative stability of foods.
out that the differences can be attributed to genetic variations, as
well as to the chromatographic conditions. Due to the complexity of
the phenolic compounds present in the samples, peak coelution is 3.8 | Determination of phytosterols and
possible during HPLC-PDA analysis. triterpenoids

Two phytosterols (β-sitosterol and cycloartenol) and one triter-


3.7 | Determination of tocopherols penoid (α-amyrin) were identified and quantified in lucuma pulp
of two varieties (Table 3). The concentrations of β-sitosterol for
Αlpha and β-tocopherol were detected in both lucuma varie- Beltrán variety (0.44 mg/100 g DW or 0.19 mg/100 g FW) and
ties, while γ-tocopherol was detected only in the Beltrán variety Seda (0.53 mg/100 g DW or 0.23 mg/100 g FW) lucuma varieties
(Table 3). Αlpha tocopherol constitutes the most abundant tocoph- were lower than those reported in mango (23.7–69.2 mg/100 g DW)
erol in both varieties, being higher in the Seda variety. The sum of (Vilela et al., 2013). Another phytosterol detected was cycloartenol,
tocopherols for the Beltrán and Seda varieties were 5.5 and 6.0 mg/ which was found in a higher concentration in Beltrán (0.35 mg/100 g
100 g DW, respectively. These values are comparable to those re- DW) than in Seda (0.25 mg/100 g DW). α-Amyrin was the triterpe-
ported in mango (1.2–9.4 mg/ 100 g DW). No δ-tocopherol was de- noid detected in both varieties of lucuma and was present in higher
tected; however, this tocopherol has only been reported in mamey concentration than the phytosterols, with of 11.85 and 12.34 μg/g
with a content of 0.36 mg/ 100 g DM (Yahia et al., 2011) and the DW for Beltrán and Seda, respectively. Its mass spectrum is shown
value was much lower than the total tocopherols found in the pre- in Figure 3. Amyrins are secondary metabolites whose bioactivity
sent work. has been widely studied and they have been attributed to antihy-
Although in comparison with other sources of tocopherols such perglycemic and anti-inflammatory properties (Vázquez, Palazon,
as oils and nuts, the total tocopherol content in lucuma is low; how- & Navarro-Ocaña, 2012). To our knowledge, this is the first report
ever, this fruit contributes a greater quantity of α-tocopherol, which of α-amyrin in P. lucuma fruits. This triterpenoid and its derivatives
is the isomer of vitamin E activity. The presence, although at low have been detected in the fruits of P. caimito and P. torta branches
concentrations of other tocopherols such as β and γ-tocopherol in (Silva et al., 2009). In this regard, Fuentealba et al. (2016) reported,
GARCÍA-RÍOS et al. |
      9 of 11

F I G U R E 3   GC-MS chromatograms
obtained for phytosterols and terpenoids
for the lucuma varieties Beltrán (a) and
Seda (b). (1) β-sitosterol, (2) α-Amyrin,
(3) Cycloartenol, (*) Not Identified,
(IS) Internal Standard. α-amyrin mass
spectrum (c)

a significant in vitro antihyperglycemic properties which could be The lipophilic AoxC is attributed to compounds such as carot-
related to this triterpenoid. enoids and tocopherols. The greater AoxC determined in the Beltrán
variety can be attributed to its higher content of carotenoids and its
greater variety of tocopherols (α, β, γ). In the studied lucuma variet-
3.9 | In vitro hydrophilic and lipophilic ies, lipophilic AoxC represents approximately 30% of the total AoxC
antioxidant capacity quantified with the ABTS method. Wu et al. (2004) reported that
the lipophilic AoxC determined with the ORAC method in 11 fruit
Significant differences (p < .05) in the lipophilic and hydrophilic AoxC species represented less than five percent of the total AoxC except
were found in the studied lucuma varieties (Table 3). Beltrán variety in watermelon (13.4%) and avocado (28.6%).
displayed the highest lipophilic and hydrophilic AoxC with values of Studies on lipophilic AoxC in other species of Pouteria are
19.3 and 8.7 μmol TE/g DW, respectively compared to Seda vari- scarce. Lipophilic AoxC values have been reported in mamey
ety (17.3 and 7.4 μmol TE/g DW). At edible ripeness, the values ob- determined with the DPPH (8.7 mg equivalent ascorbic acid
tained were higher than those reported by Fuentealba et al. (2016) (AAE)/100 g FW) and FRAP (3.5 mg AAE/100 g FW) methods
for lucuma Leiva 1 (4.8 μmol TE/g DW) that coincides with the lower that represent less than 10% of the total AoxC (Yahia et al., 2011).
TPC content also found by these authors (0.7 mg GAE/g DW) with According to what has been reported in the literature, it is not pos-
respect to the Seda and Beltrán varieties. With respect to other spe- sible to establish a conclusive relationship between the content
cies of Pouteria, such is mamey, Moo-Huchín et al. (2014) reported a of lipophilic compounds with AoxC and the value of AoxC itself
value of 15.75 μmol TE/g DW using the ABTS method. (Arnao et al., 2001).
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10 of 11       GARCÍA-RÍOS et al.

4 |  CO N C LU S I O N S Arnao, M., Cano, A., & Acosta, M. (2001). The hydrophilic and lipophilic
contribution tototal antioxidant capacity. Food Chemistry, 73, 239–
244. https://doi.org/10.1016/s0308​-8146(00)00324​-1
Both varieties of lucuma showed to be important sources of dietary Barceló, J., Nicolás, G., Sabater, B., & Sánchez, R. (2009). Fisiología Vegetal,
fiber, starch, and sugar. Moreover, they presented interesting com- 5th ed. Madrid, ES: Ediciones Pirámide.
pounds from a functional point of view. Beltrán variety presented Bechoff, A., Westby, A., Owori, C., Menya, G., Dhuique-Mayer, C.,
Dufour, D., & Tomlins, K. (2010). Effect of drying and storage on the
a higher total carotenoid content than Seda variety; however, the
degradation of total carotenoids in orange-fleshed sweetpotato cul-
carotenoid profile determined by HPLC-PDA was similar for both tivars. Journal of the Science of Food and Agriculture, 90, 622–629.
varieties. Both varieties presented linolenic acid (ω3) as the major Campos, D., Aguilar-Galvez, A., & Pedreschi, R. (2016). Stability of fruc-
fatty acid and the lipid fraction was characterized by a ratio ω6/ tooligosaccharides, sugars and colour of yacón (Smallantus sonchifo-
ω3 ≤ 0.11. Alpha-tocopherol was present in greater quantity in Seda, lius) roots during blanching and drying. International Journal of Food
Science and Technology, 51, 1177–1185.
β-tocopherol was present in the same amount in both varieties,
Chirinos, R., Campos, D., Costa, N., Arbizu, C., Pedreschi, R., & Larondelle,
while γ-tocopherol was only present in the Beltrán variety. Both va- Y. (2008). Phenolic profiles of Andean mashua (Tropaeolum tubero-
rieties presented similar amounts of phytosterols and triterpenoids, sum Ruíz & Pavón) tubers : Identification by HPLC-DAD and evalu-
highlighting the α-amyrin that might be related to the anti-inflam- ation of their antioxidant activity. Food Chemistry, 106, 1285–1298.
https://doi.org/10.1016/j.foodc​hem.2007.07.024
matory and anti-hyperglycemic activities of lucuma. The AoxC, both
Costa, T., Wondracek, C., Lopes, R., Vieira, R., & Ferreira, F. (2010).
hydrophilic and lipophilic, was higher in the Beltrán variety. In both Carotenoids Composition of canistel (Pouteria campechiana (Kunth)
varieties, lipophilic AoxC stood out, and represented approximately Baehni). Revista Brasileira de Fruticultura, 32, 903–906. https://doi.
30% of the total AoxC. These results show that lucuma fruits have org/10.1590/s0100​-29452​01000​5000083
da Costa, P., Augusto, C., Teixeira-Filho, J., & Teixeira, H. (2010).
potential for an increased and broadened industrial use.
Phytosterols and tocopherols content of pulps and nuts of Brazilian
fruits. Food Research International, 43, 1603–1606. https://doi.
AC K N OW L E D G M E N T S org/10.1016/j.foodr​es.2010.04.025
This work was supported by Vicerrectorado de Investigación— da Silva, B., Gordon, A., Jungfer, E., Marx, F., & Main, J. (2012). Antioxidant
capacity and phenolics of Pouteria macrophylla, an under-utilized
UNALM (Technological Research UNALM-2015) and Fondo Nacional
fruit from Brazilian Amazon. European Food Research and Technology,
de Desarrollo Científico, Tecnológico y de Innovación Tecnológica- 234, 761–768. https://doi.org/10.1007/s0021​7-012-1684-0
FONDECYT [grant number 124-2015-FONDECYT]. Dini, I. (2011). Flavonoid glycosides from Pouteria obovata (R. Br.) fruit
flour. Food Chemistry, 124, 884–888. https://doi.org/10.1016/j.foodc​
hem.2010.07.013
C O N FL I C T O F I N T E R E S T
Ducheteau, G., Bauer-Plank, C., Louter, A., van der Ham, M., Boerma,
The authors have declared no conflicts of interest in this article. J., van Rooijen, J., & Zandbell, P. (2002). Fast and accurate method
for total 4-desmethyl sterol(s) content in spreads, fat blends and raw
ORCID materials. Journal of the American Oil Chemists' Society, 79, 273–278.
Diego García-Ríos  https://orcid.org/0000-0002-0313-2573 https://doi.org/10.1007/s1174​6-002-0473-y
Erazo, S., Escobar, A., Olaeta, J., & Undurraga, P. (1999). Determinación
David Campos  https://orcid.org/0000-0003-1722-1187
proximal y carotenoides totales de frutos de seis selecciones de
lúcuma (Pouteria lucuma). Alimentos, 24, 67–75.
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