Food Chemistry 120 (2010) 10–14

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Food Chemistry
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Tyrosinase inhibitory activity of native plants from central Argentina: Isolation

of an active principle from Lithrea molleoides
M.E. Chiari a, M.B. Joray a, G. Ruiz b, S.M. Palacios a,1, M.C. Carpinella a,*,1
a
Fine Chemical and Natural Products Laboratory, School of Chemistry, Catholic University of Córdoba, Camino a Alta Gracia Km 10, 5000 Córdoba, Argentina
b
Herbarium Marcelino Sayago, School of Agricultural Science, Catholic University of Córdoba, Camino a Alta Gracia Km 10, 5000 Córdoba, Argentina

a r t i c l e i n f o a b s t r a c t

Article history: Screening of 91 native plants from central Argentina was carried out with the aim of finding new sources
Received 10 July 2009 of anti-tyrosinase compounds. Extracts obtained from Achyrocline satureioides, Artemisia verlotiorum,
Received in revised form 2 September 2009 Cotoneaster glaucophylla, Dalea elegans, Flourensia campestris, Jodina rhombifolia, Kageneckia lanceolata,
Accepted 16 September 2009
Lepechinia floribunda, Lepechinia meyenii, Lithrea molleoides, Porlieria microphylla, Pterocaulon alopecuroides,
Ruprechtia apetala, Senna aphylla, Sida rhombifolia, Solanum argentinum, Tagetes minuta and Thalictrum
In memory of Dr. J.J. Carpinella decipiens exhibited more than 90% inhibition of tyrosinase monophenolase activity at 1000 lg ml1.
D. elegans, L. meyenii and L. molleoides were the most potent with IC50 values of 0.48, 10.43 and 3.77
Keywords: lg ml1, respectively. D. elegans, L. molleoides and T. decipiens also showed more than 90% inhibition
Tyrosinase inhibitors of diphenolase activity at 1000 lg ml1, with the first of these being the most effective
Lithrea molleoides (IC50 = 49.27 lg ml1). (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1) was isolated from L. molleoides as an
(Z,Z)-5-(trideca-4,7-dienyl)-resorcinol effective tyrosinase inhibitor with L-tyrosine or L-DOPA as substrates (IC50 = 0.49 and 14.94 lg ml1,
Argentine plants respectively). Compound 1 was 37 times more active in monophenolase inhibitory activity than kojic acid
used as a reference. Effective extracts as well as (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol could prove to be
promising preservative agents for use in the food industry.
Ó 2009 Elsevier Ltd. All rights reserved.

1. Introduction needed in order to maintain the appearance, flavour, texture and

nutritional value of many fresh-cut products.
Tyrosinase (EC 1.14.18.1), a copper containing enzyme, also Despite the large number of PPO inhibitors, only a few of these
known as polyphenol oxidase (PPO), is involved in the initial step are used today, as many of them show side effects or low effective-
of melanin synthesis (Karioti, Protopappa, Megoulas, & Skaltsa, ness (Yi, Wu, Cao, Song, & Ma, 2009). In the past, sulphites were
2007). This protein, which catalyses the hydroxylation of L-tyrosine widely used as tyrosinase inhibitors for their antibrowning effect.
to 3,4-dihydroxyphenylalanine (L-DOPA) (monophenolase activity) However, their use has been regulated in order to ensure consumer
and the consequent oxidation of L-DOPA to dopaquinone (dipheno- safety since these preservatives affect the nutritional quality of
lase activity), is widespread in many organisms such as verte- foods and can cause allergic reactions as well as gastrointestinal
brates, invertebrates, plants and microorganisms (Chen & Kubo, distress (Ruiz-Capillas & Jiménez-Colmenero, 2009). Another cur-
2002). rently used preservative, ascorbic acid, could negatively affect
As mentioned, tyrosinase catalyses the oxidation of phenolic the aroma of some beverages and its activity is temporary
compounds into highly reactive quinones, which can polymerise (Komthong, Igura, & Shimoda, 2007). The identification of new
leading to the darkening of plant or shellfish products during pro- tyrosinase inhibitors for preservative use in food industry is thus
cessing or storage (Gawlik-Dziki, Złotek, & Świeca, 2008). Although of great concern to researchers.
enzymatic browning is beneficial for certain foods (He, Luo, & It is well known that plants are an important source of com-
Chen, 2008), it produces a less attractive appearance and loss in pounds with different activities such as insecticidal, herbicidal,
nutritional quality of a variety of fruits, vegetables and crustaceans antimicrobial, medicinal, antioxidant (Carpinella, Ferrayoli, &
(Qiu et al., 2009). Hence, agents for inhibiting this phenomenon are Palacios, 2005; Carpinella, Ferrayoli, Valladares, Defago, & Palacios,
2002; Carpinella & Rai, 2009; Carpinella et al., 2007; Rai & Carpi-
nella, 2006) and enzyme inhibition including tyrosinase (Kim &
Uyama, 2005). Many anti-tyrosinase compounds derived from
* Corresponding author. Tel.: +54 0351 4938000x611; fax: +54 0351 4938061.
E-mail addresses: ceciliacarpinella@campus1.uccor.edu.ar, cecicarpi@yahoo.com
plants are considered free of harmful side effects and can be
(M.C. Carpinella). obtained at low cost (Zheng, Cheng, Chao, Wu, & Wang, 2008).
1
Members of the National Research Council of Argentina (CONICET). These observations and the need for new agents with tyrosinase

0308-8146/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2009.09.061
 M.E. Chiari et al. / Food Chemistry 120 (2010) 10–14 11

inhibitory activity, led us to search for such chemicals among 91 2.3. Isolation of the tyrosinase inhibitor compound
extracts prepared from native plants of central Argentina that are
generally used for beverages and/or for medicinal infusions. The resulting viscous extract from L. molleoides was dissolved in
The extracts were screened with the aim of selecting those with ethanol and subjected to a column chromatography with hexane/
most activity. Subsequently, from the one obtained from Lithrea diethyl ether (Et2O)/methanol (MeOH) gradient to yield ten frac-
molleoides a compound was detected with a high anti-tyrosinase tions (F1–F10). F7–F10 were further purified by successive column
effect. chromatographies with the same solvent mixture at increasing
polarity. Finally, five fractions were obtained (F1–F5) and F2 eluted
with hexane/Et2O (60:40, v/v) was subjected to another column
2. Materials and methods chromatography and then radial preparative chromatography (sol-
vent gradient methylene chloride (CH2Cl2)/Et2O) affording with
2.1. Plant materials 100% CH2Cl2 a yellowish oil (yield 0.78 g/100 g of crushed plant
material, by HPLC).
Plants were collected in the hills of Córdoba Province, Argen- This compound was identified as the alkylresorcinol (Z,Z)-5-
tina, from November 2005 to December 2007. Voucher specimens (trideca-4,7-dienyl)-resorcinol (1) (Fig. 1) (Valcic, Wätcher, Eppler,
have been deposited in the ‘‘Marcelino Sayago” Herbarium of the & Timmermann, 2002).
School of Agricultural Science, Catholic University of Córdoba and (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1): C19H28O2. tR 4.46 min
were authenticated by the botanist, Gustavo Ruiz. (by HPLC). EI-MS m/z (Int. rel.%) 288 [M+] (3.3), 287 [M+-1] (4.0),
Plants were selected according to their availability, accessibility 217 [M-71] (2.7), 203 [M-85] (2.3), 163 [M-125] (14.6), 124 [M-
and especially the lack of scientific information about their activity 164] (100), 81 (11.1), 77 (14.4). 1H NMR (400 MHz, CDCl3) d 6.23
and/or chemical pattern. (1 H, d, J = 2.0 Hz, H-4,6), 6.17 (1 H, t, J = 2.0 Hz, H-2), 5.39 (1 H,
Crushed aerial plant material was extracted by 48 h maceration m, H-40 ), 5.38 (1 H, m, H-80 ), 5.36 (1 H, m, H-50 ), 5.34 (1 H, m, H-
with ethanol. The yields of the most active extracts, obtained after 70 ), 2.76 (2 H, t, J = 6.0 Hz, H-60 ), 2.51 (2 H, t, J = 7.8 Hz, H-10 ),
solvent removal and expressed as percentage weight of air-dried 2.10 (2 H, q, J = 6.9 Hz, H-30 ), 2.03 (2 H, q, J = 7.0 Hz, H-90 ), 1.65 (2
crushed plant material, are shown in Table 1. H, q, J = 7.7 Hz, H-20 ), 1.33 (2 H, m, H-100 ), 1.30 (2 H, m, H-120 ),
1.28 (2 H, m, H-110 ), 0.91 (3 H, t, J = 6.8 Hz, Me). 13C NMR
(100.0 MHz, CDCl3) d 14.2 (C-130 ), 22.6 (C-120 ), 25.7 (C-60 ), 26.7
2.2. Chemicals, equipment and reagents
(C-30 ), 27.2 (C-90 ), 29.3 (C-100 ), 30.9 (C-20 ), 31.5 (C-110 ), 35.1
(C-10 ), 100.2 (C-2), 107.9 (C-4,6), 127.8 (C-70 ), 128.6 (C-50 ), 129.1
L-Tyrosine, 3,4-dihydroxy-L-phenylalanine (L-DOPA) and lyoph-
(C-40 ), 130.4 (C-80 ), 145.6 (C-5), 156.8 (C-1,3).
ilised mushroom tyrosinase were purchased from Sigma–Aldrich
CO (St. Louis, MO). Kojic acid was obtained from Merck (Darmstadt,
Germany). Silica gel (70–230 mesh) used for column chromatogra- 2.4. Tyrosinase inhibitory assay
phy was purchased from Sigma–Aldrich CO and all solvents were
HPLC grade. 1H- and 13C-NMR spectra were recorded in Chloro- Tyrosinase inhibitory activity was determined spectrophoto-
form-d3 with Bruker AVANCE II 400 spectrometer (Bruker Corpora- metrically. First, 2 ll of mushroom tyrosinase (2500 U ml1 in
tion, Ettlingen, Germany) operated at 400 MHz for 1H and at
100 MHz for the 13C nucleus. Chemical shifts (parts per million)
are relative to internal tetramethylsilane used as a reference
OH
(d = 0.00). MS spectra were measured with a ZAB SEQ (BeqQ)
instrument (VG Analytical, Manchester). For quantifying the pure 2
compound, HPLC was performed on a Phenomenex Prodigy 5 l 7´ 4´
ODS (4.6 mm i.d.  250 mm) reversed-phase column eluting with OH
13´ 1´ 4
90% acetonitrile in water with 1% trifluoracetic acid (TFA) as mobile
phase and UV detection at 280 nm. Fig. 1. Chemical structure of (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1).

Table 1
Native plants from central Argentina showing high tyrosinase inhibitory activity.

Plant species Family Common name Yield (%) Statusa Voucher: UCCOR number
Achyrocline satureioides (Lam.) DC. Asteraceae marcela hembra 4.5 N 140
Artemisia verlotiorum Lamotte Asteraceae yuyo de San Vicente 3.5 Adv. 230
Cotoneaster glaucophylla Franch. Rosaceae crategus 9.1 I 126
Dalea elegans Hook. and Arn. Fabaceae 11.8 N 254
Flourensia campestris Griseb. Asteraceae chilca 11.8 N 221
Jodina rhombifolia (Hook. and Arn.) Reissek Santalaceae sombra de toro 5.0 N 153
Kageneckia lanceolata Ruiz and Pav. Rosaceae durazno de la sierra 14.7 N 264
Lepechinia floribunda (Benth.) Epling Lamiaceae salvia blanca 3.7 N 195
Lepechinia meyenii (Walp.) Epling Lamiaceae 3.8 N 233
Lithrea molleoides (Vell.) Engl. Anacardiaceae molle de beber 7.2 N 183
Porlieria microphylla (Baill.) Descole, O’Donell and Lourteig Zygophyllaceae cucharero 1.1 N 154
Pterocaulon alopecuroides (Lam.) DC. Asteraceae 5.5 N 217
Ruprechtia apetala Wedd. Polygonaceae manzano del campo 1.9 N 151
Senna aphylla (Cav.) H.S. Irwin et Barneby Fabaceae pichana 5.1 N 174
Sida rhombifolia L. Malvaceae escoba dura 2.2 N 141
Solanum argentinum Bitter and Lillo Solanaceae duraznillo blanco 5.4 N 34
Tagetes minuta L. Asteraceae suico 2.5 N 138
Thalictrum decipiens Boivin Ranunculaceae albaquilla 4.7 N 287
a
Adv.: adventive; I: introduced; N: native.
12 M.E. Chiari et al. / Food Chemistry 120 (2010) 10–14

50 mM phosphate buffer, pH 6.5) was mixed with 138 ll of 50 mM Table 3

phosphate buffer. Then, 20 ll of tested solution dissolved in DMSO Most effective extracts of native plants from central Argentina on mushroom
tyrosinase. Diphenolase activity.
at the concentrations needed or DMSO (control), were added. All
samples were first tested at 1000 lg ml1 and those showing 90% Species Inhibition IC50 (lg ml1) values
inhibition or higher in three different repetitions were further eval- (%)a,b and 95%
confidence limits
uated for the concentration necessary for 50% inhibition (IC50). The (lower, upper)b
assay mixture was then incubated at 37 °C for 90 min with gentle
Dalea elegans 99.34 ± 1.11 49.27 (23.16, 104.80)
agitation. At this stage a stable absorbance was reached. Finally, Lithrea molleoides 92.33 ± 0.55 79.44 (27.48, 229.67)
40 ll of 2.5 mM L-tyrosine or L-DOPA in phosphate buffer were Thalictrum decipiens 93.02 ± 0.12 158.03 (76.37, 326.99)
added and immediately monitored (t = 0) at 450 nm for dopa- (Z.Z)-5-(trideca-4. 98.35 ± 1.17 14.94 (5.85, 38.09)
chrome formation in the reaction mixture. Measurements were re- 7-dienyl)-resorcinol
Kojic acid 98.66 ± 0.48 2.64 (1.06, 6.57)
peated up to 15 min for monophenolase activity and up to 5 min
for diphenolase activity. Differences in absorbance between each a
Data represent the mean ± SD of the parameter evaluated.
b
time measured and time zero were calculated and the inhibition Value at 2 min from the beginning.
percentage was determined respect to control.
Kojic acid, dissolved in 50 mM phosphate buffer was used as a
positive control. Each measurement was made at least in duplicate. lowed by L. molleoides (IC50 = 79.44 lg ml1) and finally T. decipiens
(IC50 = 158.03 lg ml1) (Table 3). Because of these results and due
to its high availability and accessibility in the hill area of Córdoba,
2.5. Statistical analysis
L. molleoides was selected as a first potential source of new anti-
tyrosinase compounds.
The results are expressed as mean ± standard error (SD). The
From the ethanolic extract of this plant, a compound containing
inhibitory concentration (IC50) was calculated by log-Probit
a C-13 unsaturated hydrocarbon side chain attached at position
analysis.
five to a dihydroxyphenyl moiety was obtained. It was identified
as (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1) (Fig. 1).
3. Results Compound 1 inhibited the monophenolase activity of tyrosi-
nase showing an IC50 value of 0.49 lg ml1 (Table 2), while 50%
Because of the urgent need for new agents with inhibitory activ- inhibition of the diphenolase activity was reached at 14.94 lg ml1
ity on tyrosinase, we performed a screening of extracts obtained (Table 3). It inhibited tyrosinase activity when using L-tyrosine or
from native plants from central Argentina with the aim of finding L-DOPA as substrates in a dose-dependent manner (Fig. 2a and
these. b). It exhibited significant inhibitory effect on monophenolase
The results of the screening showed that 18 of the 91 extracts activity, with 32% inhibition at 0.24 lg ml1, 93% at 3.91 lg ml1
tested showed high monophenolase inhibitory activity on tyrosi- and total inhibition at 7.81 lg ml1. At the latter concentration,
nase with percentages above 90% at 1000 lg ml1 (see Table 2). kojic acid showed a weak inhibition (22%) and just reached 96%
Dalea elegans was the most potent plant showing an IC50 of inhibition at 125 lg ml1. Compound 1 inhibited 21% of dipheno-
0.48 lg ml1 followed by L. molleoides (IC50 = 3.77 lg ml1) and lase enzyme activity at 3.91 lg ml1, 81% at 62.50 lg ml1 and
Lepechinia meyenii (IC50 = 10.43 lg ml1) (Table 2). 98% at 1000 lg ml1. In regard to this activity, kojic acid was more
D. elegans, L. molleoides and Thalictrum decipiens showed more effective than the alkyl resorcinol, reaching 98% inhibition at
than 90% inhibition of diphenolase activity at 1000 lg ml1, with 250 lg ml1.
the first of these being the most effective (IC50 = 49.27 lg ml1) fol- Monophenolase activity is typically characterised by a lag time,
which is the time required to reach the steady-state concentration
of o-diphenols which reduce the met form of tyrosinase to the
Table 2
Most effective extracts of native plants from central Argentina on mushroom
deoxy form which then converts to the oxy form. The kinetic course
tyrosinase. Monophenolase activity. of the oxidation of L-tyrosine observed with the formation of dopa-
chrome in the presence of different concentrations of compound 1
Species Inhibition IC50 (lg ml1) values and 95%
(%)a,b confidence limits (lower, upper)b
is shown in Fig. 3. At concentrations of 1.95 and 3.91 lg ml1, the
lag period was extended till 10 min. Furthermore, compound 1 re-
Achyrocline satureioides 95.42 ± 2.15 26.72 (10.39, 68.71)
Artemisia verlotiorum 94.88 ± 1.94 79.69 (17.11, 37.11)
duced the reaction rate in a dose-dependent manner.
Cotoneaster glaucophylla 94.67 ± 0.25 19.17 (9.55, 38.48)
Dalea elegans 90.75 ± 1.36 0.48 (0.17, 1.36)
Flourensia campestris 100.00 ± 0 107.91 (65.28, 178.37)
4. Discussion
Jodina rhombifolia 95.90 ± 0.55 31.35 (6.67, 147.35)
Kageneckia lanceolata 93.15 ± 1.97 26.87 (11.66, 61.94)
Lepechinia floribunda 95.91 ± 0.37 266.19 (115.42, 613.91) Much effort has been spent in the search for new effective, sta-
Lepechinia meyenii 96.99 ± 2.33 10.43 (4.45, 24.42) ble, low-toxic anti-tyrosinase compounds.
Lithrea molleoides 99.56 ± 1.07 3.77 (2.40, 5.92) According to IC50 values, using L-tyrosine as substrate, we were
Porlieria microphylla 97.40 ± 2.83 45.03 (21.36, 94.95)
surprised to find that ethanolic extract from D. elegans
Pterocaulon alopecuroides 99.36 ± 1.57 14.28 (5.79, 35.23)
Ruprechtia apetala 91.54 ± 0.43 118.97 (54.29, 260.67) (IC50 = 0.48 lg ml1) was 38 times more effective than that of kojic
Senna aphylla 97.52 ± 0.57 76.39 (38.66, 150.92) acid, the IC50 of which was 18.25 lg ml1. L. meyenii and L. molleo-
Sida rhombifolia 94.99 ± 1.17 103.36 (58.72, 181.94) ides extracts (IC50 = 10.43 and 3.77 lg ml1, respectively) were also
Solanum argentinum 98.04 ± 0.67 19.97 (3.10, 128.88)
highly active showing almost two and five times, respectively,
Tagetes minuta 97.20 ± 0.58 41.58 (14.94, 115.74)
Thalictrum decipiens 92.21 ± 2.66 21.90 (8.12, 59.23)
more potency than that of the reference. Cotoneaster glaucophylla,
(Z.Z)-5-(trideca-4,7- 100 ± 0 0.49 (0.22, 1.09) Pterocaulon alopecuroides, Solanum argentinum and T. decipiens
dienyl)-resorcinol (IC50 = 14.28–21.90 lg ml1), in turn, exhibited similar levels of
Kojic acid 100 ± 0 18.25 (9.37, 35.53) effectiveness to that of kojic acid.
a
Data represent the mean ± standard error media of the evaluated parameter. Among plants with inhibition of the oxidation of L-DOPA cata-
b
Value at 10 min from the beginning. lysed by tyrosinase, D. elegans, L. molleoides and T. decipiens
 M.E. Chiari et al. / Food Chemistry 120 (2010) 10–14 13

Fig. 2. (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1) inhibits tyrosinase activity in a dose-dependent manner. Compound 1 or kojic acid were incubated with tyrosinase at 37 °C
with (a) L-tyrosine or (b) L-DOPA as substrates. Data represent the mean ± SD for duplicate within one experiment as a percentage of the control mean measured at 10 and
2 min from beginning of experiment for mono- and diphenolase activity, respectively.

kai, 2000). Among these, 4-hexylresorcinol has been recognised as

a safe and effective tyrosinase inhibitor for use in the food industry
(Zheng et al., 2008) and its use as a preservative is permitted in
many countries (Martínez-Alvarez, López-Caballero, Montero, &
Gómez-Guillén, 2007). According to Kim and Uyama (2005), the
effectiveness of these molecules is due to substitution in 4-posi-
tion, mainly with hydrophobic substituents, since resorcinol with-
out substitution is a poor inhibitor of the phenoloxidase (Kim &
Uyama, 2005). The same seems to happen with resorcinols substi-
tuted in 5-position, as is demonstrated by compound 1 activity and
for other 5-alkyl resorcinols (Miura et al., 1995). The long hydro-
phobic alkyl chain may associate with the hydrophobic binding
pocket close to the binuclear copper active site, as happened with
Fig. 3. Course of the oxidation of L-tyrosine by tyrosinase in the presence of other hydrophobic inhibitors (Conrad, Dawso, Hubbard, Meyers, &
different concentrations of (Z,Z)-5-(trideca-4,7-dienyl)-resorcinol (1). Concentra- Strothkamp, 1994).
tions of 1 for curves 1–6 were 0, 0.24, 0.49, 0.98, 1.95 and 3.91 lg ml1, Despite L. molleoides shows medicinal properties (Araujo, Bela,
respectively. Compound 1 was incubated with tyrosinase at 37 °C with L-tyrosine
Bueno, Rodrigues, & Shimizu, 2006; Fernández et al., 2002; Kott
as substrate. Dopachrome formation was determined at different times till 15 min.
et al., 1999; Muschietti et al., 2005), its effect in inhibiting tyrosi-
nase has not yet been explored. It has been reported that com-
(IC50 = 49.27–158.03 lg ml1) showed themselves less effective pound 1 produces paralysis on nematodes (Valcic et al., 2002)
than that of the positive control (IC50 = 2.64 lg ml1). and exerts a cytotoxic effect on human tumoral cell lines (López
A literature survey appears to show no information of anti- et al., 2005), but its tyrosinase activity inhibition has not yet been
tyrosinase properties in any of the 18 plant species with high described. This means that this is the first time that a compound
inhibitory effectiveness. showing this property has been isolated from L. molleoides.
A compound belonging to the alkylresorcinol family was iso- Although toxic effects of compound 1 have not yet been deter-
lated from L. molleoides and identified as (Z,Z)-5-(trideca-4,7-die- mined, other alkyl resorcinols can be found in many edible plants,
nyl)-resorcinol 1. Surprisingly, compound 1 (IC50 = 0.49 lg ml1) fruits and grains (Seitz, 1992), thus denoting its low toxicity. Be-
was 37 times more active than kojic acid when L-tyrosine was used sides, L. molleoides is used in traditional medicine as an infusion
as substrate. Its activity was 81 times stronger than the well- (López et al., 2005), and thus few or no side effects are expected.
known tyrosinase inhibitor arbutin, the IC50 of which corresponded In this sense, it is expected that compound 1 could present few side
to 39.71 lg ml1 (Zheng et al., 2008) and 44 times more active than effects, thus ensuring its safe use as an anti-tyrosinase compound.
the extensively used sodium bisulphite (IC50 = 21.84 lg ml1) From the more than satisfactory results obtained from this work
(Friedman & Bautista, 1995). and taking into account their potential applications in food indus-
The well-known trans-stilbene resveratrol, showed an IC50 of try, the tyrosinase inhibitors found could arise as promising agents
12.46 lg ml1 (Kim, Yun, Lee, Lee, & Min, 2002) which is higher for preserving many food products.
than that observed for compound 1. Oxyresveratrol isolated from
Morus alba and considered one of the most potent tyrosinase inhib-
Acknowledgements
itors in nature (Kim, Son, Chang, Kang, & Kim, 2003), showed an
IC50 of 0.29 lg ml1 (Kim et al., 2002), very close to that obtained
This work was supported by the Catholic University of Córdoba,
for the isolated alkylresorcinol when inhibiting the hydroxylation
FONCyT PICT 2005 and PICTO CRUP 2005. MBJ gratefully acknowl-
of L-tyrosine.
edges receipt of a fellowship from CONICET. We thank Joss Hey-
When L-DOPA was used as substrate, compound 1 showed sig-
wood for revising the English language.
nificant inhibition of diphenolase activity (IC50 of 14.94 lg ml1)
but weaker in comparison to that of kojic acid. On the contrary,
the alkylresorcinol was 1.6 times more active than ascorbic acid References
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Compounds with 4-substituted resorcinol skeleton have been Anti-ulcerogenic activity of the aerial parts of Lithraea molleoides. Fitoterapia, 77,
reported as potent inhibitors of tyrosinase (Shimizu, Kondo, & Sa- 406–407.
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