JOURNAL OF FUNCTIONAL FOODS 4 (2 0 1 2) 3 3 934 7

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Polyphenols, carotenoids, and ascorbic acid in underutilized

medicinal vegetables

Nuri Andarwulana,b,*, Dewi Kurniasihb, Riza Aris Apriadyb, Hardianzah Rahmatb,

Anna V. Rotoc, Bradley W. Bollingc
a
Southeast Asian Food and Agricultural Science and Technology (SEAFAST) Center, Bogor Agricultural University, Bogor, Indonesia
b
Department of Food Science and Technology, Bogor Agricultural University, Bogor, Indonesia
c
Department of Nutritional Sciences, University of Connecticut, Storrs, CT, USA

A R T I C L E I N F O A B S T R A C T

Article history: The polyphenols, carotenoids, ascorbic acid, and protein were determined in 24 underuti-
Received 9 September 2011 lized medicinal vegetables from Indonesia. Anacardium occidentale, Sauropus androgynus (L.)
Received in revised form Merr., and Moringa pterygosperma Gaertn. leaves were rich sources of flavonoids, with
27 December 2011 118144 mg/100 g fresh weight. Quercetin, kaempferol, and chlorogenic acid were the
Accepted 5 January 2012 predominant polyphenols among those measured in vegetables. Polyscias pinnata leaves
Available online 30 January 2012 and Solanum torvum Swartz fruits had the most phenolic acids, with 53 and 36 mg/100 g,
respectively. Moringa pterygosperma had the most carotenoids among vegetables, with
Keywords: 14 mg b-carotene equivalents (bCE)/100 g. Ascorbic acid content of fresh vegetables was
Medicinal vegetable 12.03494.43 mg/100 g. A. occidentale, S. androgynus, Ocimum americanum L., Cosmos caudatus
Phenolic acid H.B.K., and Carica papaya L. (papaya) leaves had more than 100 mg ascorbic acid/100 g.
Flavonoid Thus, a number of underutilized vegetables from Indonesia may be rich sources of func-
b-Carotene
tional components including polyphenols and ascorbic acid.
Ascorbic acid
 2012 Elsevier Ltd. All rights reserved.

1. Introduction Among functional components, polyphenols are particu-

Vegetables may confer a variety of health benefits. Increased
vegetable consumption is correlated with reduced risks of
cardiovascular disease, stroke, arthritis, inflammatory bowel
diseases, and some cancers (Amre et al., 2007; Dauchet
et al., 2010; Griep, Verschuren, Kromhout, Ocke, & Geleijnse,
2011; Jain, Hislop, Howe, & Ghadirian, 1999). Many underuti-
lized vegetables are also used medicinally (Andarwulan,
Batari, Sandrasari, Bolling, & Wijaya, 2010). Vitamins, miner-
als, dietary fiber, and phytochemicals contribute to the
functionality of vegetables. Characterizing the distribution
of these functional components can inform dietary and
horticultural efforts to increase intake of these potentially
beneficial components from underutilized vegetables.
larly promising for their role in health-promotion. Polyphe-
nols or their metabolites modulate gene expression,
epigenetic regulation, cell signaling, inflammation, antioxi-
dant function, detoxification, and immune function (Bolling,
Ji, Lee, & Parkin, 2011; Kang, Shin, Lee, & Lee, 2011; Yun, Jialal,
& Devaraj, 2010). Vegetables contain a number of polyphenol
classes, including hydroxycinnamic and hydroxybenzoic
acids, flavonols, flavan-3-ols, flavones, flavanones, anthocya-
nins, phenolic aldehydes, stilbenes, hydrolyzable tannins,
and proanthocyanidins, among others (Naczk & Shahidi,
2006; Shahidi, Chandrasekara, & Zhong, 2011).
Ascorbic acid and carotenoids are also important func-
tional vegetable components. Ascorbic acid is an antioxidant
vitamin that acts synergistically with tocopherol to preserve

* Corresponding author at: Southeast Asian Food and Agricultural Science and Technology (SEAFAST) Center, Bogor Agricultural
University, Jl Puspa No. 1, Kampus, IPB Darmaga, Bogor, Indonesia. Tel./fax: +62 251 8629903.
E-mail address: nuri@seafast.org (N. Andarwulan).
1756-4646/$ - see front matter  2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jff.2012.01.003
340 JOURNAL OF FUNCTIONAL FOODS
antioxidant function in chronic disease states (Bruno et al.,
2006; Traber & Stevens, 2011). Vegetable carotenoids have
antioxidant functions and can prevent vitamin A deficiencies
when consumed with lipids to increase their bioavailability
(Ribaya-Mercado et al., 2007; Shahidi & Zhong, 2010). This is
particularly important in developing countries at risk for vita-
min A deficiency (Tan et al., 2002).
Polyphenols, ascorbic acid, and carotenoids have not been
adequately characterized in a number of underutilized vegeta-
bles, particularly in Indonesia. We previously reported that
Indonesian medicinal vegetables contain the flavonols querce-
tin, kaempferol, and myricetin and the flavones luteolin and
apigenin, and possessed antioxidant activity (Andarwulan
et al., 2010). Particularly, Sauropus androgynus (L.) Merr. (katuk),
Cosmos caudatus H.B.K. (kenikir), and Polyscias pinnata (kedon-
dong cina) were identified as rich sources of dietary flavonoids
(Andarwulan et al., 2010). The aim of this study was to provide a
more comprehensive characterization of polyphenols, ascor-
bic acid, carotenoids, and protein in underutilized Indonesian
vegetables. Therefore, we quantified flavonols, flavones, phe-
nolic acids, anthocyanins, total phenols, b-carotene, ascorbic
acid, and protein in 24 Indonesian vegetables. This study is sig-
nificant in that it is apparently the first report of polyphenols in
a number of these vegetables.
2. Materials and methods
Butylated hydroxyanisole (BHA), tert-butylhydroquinone
(TBHQ), gallic acid, caffeic acid, ferulic acid, chlorogenic acid,
and b-carotene were from SigmaAldrich (St. Louis, MO, USA).
Methanol, acetic acid, chloroform, acetonitrile, ethanol, HPLC
grade water, HPLC grade methanol, FolinCiocalteu reagent,
Na2CO3, KH2PO4, HCl, soluble starch, KI, and iodine were from
Merck (Darmstadt, Germany). Hexane and acetone were from
Brataco Chemica (Bandung, Indonesia), KOH was from BDH
(Leicestershire, UK).
The vegetables sampled in the present study represent a
new sampling and analysis from our previous report
(Andarwulan et al., 2010). Samples of 0.51.0 kg of fresh
vegetables free from obvious defects were obtained in Bogor,
Indonesia (Table 1). C. caudatus H.B.K. leaves, Etlingera elatior
(Jack) R.M.Sm. flowers, Ocimum americanum L. leaves, S.
androgynus leaves, Pilea melastomoides (Poir.) Bl. leaves, Talinum
triangulare (Jacq.) Willd. leaves, Allium schoenoprasum L., Sola-
num torvum Swartz fruits, Vigna unguiculata (L.) Walp. leaves,
Saccharum edule Hassk flowers, Sechium edule (Jacq.) Swartz
leaves, Carica papaya L. flowers, Anacardium occidentale L.
leaves, and Arcypteris irregularis (C.Presl) Ching leaves were
obtained from the Bogor traditional market, while the rest
were harvested from uncultivated fields near Bogor Agricul-
tural University. The vegetables were identified and classified
by Dr. Eko Baroto Waluyo, APU, Indonesian Institute of Sci-
ence, Research Center for Biology. Vegetables were washed,
drained, cut into pieces, and frozen at 20 C. Within 12 h
of freezing, samples were lyophilized for at least 48 h, pow-
dered, passed through 30 mesh sieves, and stored at 20 C
until further analysis.
The total phenols content of extracts was determined
using a modified method of Singleton, Orthofer, and
4 ( 2 0 1 2 ) 3 3 9 3 4 7
Lamuela-Ravent (1999) as previously described (Andarwulan
et al., 2010). Total phenols contents were expressed as gallic
acid equivalents. For anthocyanin analysis, lyophilized vege-
table powder, was extracted 1:10 (w/v) with 10 mL of 5% HCl
in water at 4 C overnight (Raharja & Dianawati, 1995). The
mixture was filtered and analyzed by the pH differential meth-
od (Lees & Francis, 1982).
Flavonoids were extracted and quantified by a method
by Hertog et al. (1995), with modifications (Andarwulan
et al., 2010). Briefly, lyophilized vegetable powder was incu-
bated with acidified aqueous methanol and TBHQ to hydro-
lyze and release bound flavonoids. The resulting extract
was analyzed with a Shimadzu HPLC with UV detection
(Kyoto, Japan) and a Develosil ODS column (Nomura Chem-
ical, Seto, Japan). Flavonoids were separated with an aceto-
nitrile gradient in phosphate buffer, tentatively identified
based on UV absorbance and retention time, and quantified
using dilutions of authentic standards. A representative
chromatogram for flavonoid analysis is in Fig. 1. On column
limits of detection were 0.78 pg myricetin, 1.12 pg luteolin,
0.56 pg quercetin, 4.4 pg apigenin, 0.94 kaempferol, deter-
mined experimentally as three times the standard deviation
of the area under the curve for each compound (Rounds &
Nielsen, 2000). Limits of quantification were >10 times the
standard deviation of the area under the curve for each
compound.
Extraction and quantification of phenolic acids was
performed by acid hydrolysis and release of bound phenolics
according to the method described by Mattila and
Kumpulainen (2002). Lyophilized vegetable powder (0.5 g) was
reconstituted in 7 mL of 62.5% methanol in 15% acetic acid in
water with 2 g/L BHA, sonicated for 30 min (Branson ultrasonic
3510, Danbury, CT, USA), and made to 10 mL with distilled
water. Extract was filtered through a 0.45 lm membrane, and
20 lL filtrate was injected to the HPLC and column used for
flavonoid analysis, with the following modifications. Isocratic
methanol:0.4% acetic acid (80:20, v/v) at 1 mL/min was used
to elute phenolic acids with detection at 290 nm (Singh et al.,
2008). Phenolic acids were tentatively identified based on UV
absorbance and retention time, and quantified using standard
curves of authentic compounds. A representative chromato-
gram for phenolic acid analysis is in Fig. 1. Limits of detection
and quantification were determined as described above. Limits
of detection were 16 pg ferulic acid, 19.4 pg chlorogenic acid,
and 16.6 pg caffeic acid on column. Additional acid and base
hydrolysis of extraction residues did not yield additional
phenolic acids.
Total carotenoids and b-carotene were determined after
saponification and extraction (Zakaria-Rungkat, Djaelani,
Setiana, & Nurrochmah, 2000). Hexane and acetone (1:1, v/v)
was used to extract 0.25 g lyophilized vegetable powder (20:1,
v/w) thrice and passed through Whatman 42 filter paper. Sol-
vent was removed by rotary evaporation at 45 C. Extract was
mixed with 4 mL of 5% KOH in methanol, sonicated, and incu-
bated at 70 C for 30 min. The extracts were cooled and mixed
with 4 mL deionized water and 8 mL hexane. Following centri-
fugation, the organic phase was withdrawn, and the aqueous
phase reextracted with 6 mL hexane and 3 mL of 5% acetic
acid. Organic phases were combined, dried by rotary evapora-
tion at 45 C, and reconstituted in 4 mL hexane. Absorbance
 JOURNAL OF FUNCTIONAL FOODS 4 ( 20 1 2) 3 3 934 7 341

Table 1 Underutilized vegetables of Indonesian origin and their uses analyzed in the present study.
Species; edible part Common names Reported medicinal or traditional usesa

Allium schoenoprasum L.; all parts
Anacardium occidentale L. leaf
Arcypteris irregularis (C.Presl) Ching; leaf
Carica papaya L.; flower
Centelia asiatica (L.) Urb.; all parts
Cosmos caudatus H.B.K.; leaf
Etlingera elatior (Jack) R.M.Sm.; flower
Hydrocotyle sibthorpioides Lmk.; all parts
Morinda citrifolia L.; leaf
Moringa pterygosperma Gaertn.; leaf
Nothopanax scutellarius (Burm.f.) Merr.; leaf
Ocimum americanum L.; leaf
Pilea melastomoides (Poir.) Bl.; leaf
Pluchea indica (L.) Less.; leaf
Polyscias pinnata; leaf
Polyscias scutellaria (Burm.f.) Fosb.; leaf
Portulaca oleracea L.; leaf and stem
Saccharum edule Hassk.; flower
Sauropus androgynus (L.) Merr.; leaf
Sechium edule (Jacq.) Swartz; leaf
Sesbania grandiflora (L.) Pers.; flower
Solanum torvum Swartz; fruit
Talinum triangulare (Jacq.) Willd.; leaf
Vigna unguiculata (L.) Walp.; leaf
a
References for these functions are provided in Supplemental Table 1.
kucai, wild chives
jambu mete, cashew
pakis, fern
pepaya, papaya
antanan, Indian pennywort
kenikir, wild cosmos
kecombrang, torch ginger
antanan beurit, lawn
marshpennywort
mengkudu, Indian mulberry
kelor, horseradish tree
mangkokan
kemangi, basil
pohpohan
beluntas, Indian camphorweed
kedondong cina, balfour aralia
mangkokan putih, shield aralia
krokot, little hogweed, purslane
terubuk, vegetable cane
katuk, chekkurmanis
labu siam, chayote
turi, vegetable hummingbird
takokak, turkey berry
daun ginseng, Ceylon spinach
lembayung, blackeyed pea
Antihypertensive, intestinal antibiotic, blood thinner
Anti-rheumatic
Anthyhypertensive, tonsils, anti-rheumatic
Antipyretic, antimalarial
Diuretic, antihypertensive; memory enhancer, ergogenic
aid for elderly, blood detoxicant; for hemorrhoids, liver
disease, antitusive, sore throat, asthma, colitis, kidney
stones, and Celiac disease
Appetite stimulant, bone strengthener, insect repellant,
for weak stomachs
Food antibacterial, deodorant
Memory enhancer, antipyretic, anti-inflammatory,
diuretic, antibacterial, insecticide, antihistamine; for
blood detoxification, circulation, bleeding
Anthelmintic
Antihypertensive, anti-diarrheic for diabetes mellitus
and heart disease
Lactation aid; for breast edema, hair loss, urination
For cough, skin diseases, and rheumatism; treat
headache, conjunctivitis, malaria, and fragrance
Appetite stimulant, anti-diaphoretic, antipyretic,
digestive aid, deodorant, antibacterial, anti-diarrheal,
antitusive, emollient
Deodorant, appetite suppressant, eyewash
Diuretic, anti-diaphoretic, deodorant, anti-alopecia,
hairloss
Anti-diarrheal, anti-inflammatory, anthelmintic,
laxative; for appendix, breast inflammation,
hemorrhoids
Lactation aid, antipyretic, colorant
Antihypertensive, anti-coagulant; for arteriosclerosis,
kidney stones, respiratory and digestive systems,
circulation
For headache, as eyedrop
Anti-edemic, analgesic, circulation improvement,
alleviate pain, antitusive, anti-inflammatory; for
stomach pain, toothache, cataracts, menstruation
disorders, hemorrhoids, sore breasts, influenza,
swelling, ulcers, soreness, sore waist, high uric acid,
bone loss, heart palpitations, and detoxification
For circulation, mucous membranes, stimulate mucous
membranes, anti-edemic, antibacterial, anti-viral,
appetite stimulant
Dermatitis and swellings

was measured at 450 nm, with hexane as a blank. Total
carotenoids (lg/g) were determined as: Absorbance at
450 nm [10 lg/lL/E1%] [4000 lL/mass powdered vegetable
(g)] dilution factor, where E1% is was the extinction value
of a 1% b-carotene solution at 450 nm (=2600) and DF was
the dilution factor. For b-carotene quantification, total carot-
enoid extracts were dried by rotary evaporation, dissolved in
5% chloroform in methanol, and kept at 20 C overnight.
Solutions were then filtered with 0.22 lm membrane, dried,
and dissolved in 2 mL mobile phase. Extract (20 lL) was ana-
lyzed by HPLC-UV with a Eclipse XDB-C18 column (Agilent,
Santa Clara, CA, USA) and isocratic mobile phase of
methanol:acetonitrile:chloroform (48.5:48.5:3.0, v/v/v) at
0.8 mL/min and detection of b-carotene at 450 nm.
Ascorbic acid was determined in fresh vegetables homog-
enized in water (2:1, w/v) by titration (Jacobs, 1951). Water and
protein content were determined by AOAC (1984, 1995) meth-
ods 930.04 and 9260.5, respectively.
Data are reflective of two independent experiments with
duplicate analysis. Analysis of variance, Duncans test, and
Tukeys test (P < 0.05) were used to determine statistical
significance, using SPSS 13.0 (IBM Corp., New York, NY,
USA). Relationships between phenolic and non-phenolic
compounds were determined by Principal Component
342 JOURNAL OF FUNCTIONAL FOODS 4 ( 2 0 1 2 ) 3 3 9 3 4 7

3. Results and discussion

Vegetables contain a wide variety of both essential and non-

Fig. 1 Representative chromatograms of HPLC analysis of
vegetable extracts for (A) H. sibthorpioides flavonoids; and (B)
P. scutellaria phenolic acids.
Analysis (PCA) using Minitab 16 (Minitab Inc., State College,
PA, USA).
essential nutrients. The relative abundance of these nutrients
determines the contribution of specific vegetables to prevent
deficiencies, optimize health or performance, or prevent dis-
eases. Many nutrients from Western foods have been charac-
terized and quantified. However similar analysis of vegetables
from Indonesia is lacking. Many vegetables have traditional
medicinal uses in Indonesia, particularly among the Javanese
population. We hypothesized these vegetables would be
sources of carotenoids, ascorbic acid, polyphenols, and
proteins. Therefore we quantified these components in 24
underutilized vegetables of Indonesian origin (Table 1).
The contents of flavonoids quercetin, kaempferol, myrice-
tin, luteolin, and apigenin of vegetables ranged from to
0.30 mg/100 g fresh weight (fw) for Portulaca oleracea L. to
143.58 mg/100 g fw for A. occidentale L. leaves (Table 2).
Together, quercetin and kaempferol constituted more than
60% of the sum of vegetable flavonoids. Quercetin was the only
flavonoid detected in E. elatior, P. oleracea, and Sa. edule. No
flavones were detected in leaves or flowers of T. triangulare,
S. androgynus, P. pinnata, C. caudatus, Nothopanax scutellarius
(Burm.f.) Merr., Morinda citrifolia L., A. irregularis, and Sesbania
grandiflora (LOD 5.6 lg luteolin, 22 lg apigenin per gram
dry vegetable matter). A. occidentale, S. androgynus, and M.
pterygosperma Gaertn. leaves had more than double the
flavonoid content of other vegetables. A. occidentale and M.
pterygosperma flavonoid content were 87% and 81% quercetin,
respectively. In contrast, S. androgynus flavonoids were 97%
kaempferol. Se. edule (Jacq.) Swartz leaves and A. occidentale

Table 2 Flavonoid content of underutilized vegetables of Indonesian origin.

Vegetable Flavonoids (mg/100 g fw)
Myricetin Luteolin Quercetin Apigenin Kaempferol Total

A. occidentale 8.28 0.17 nd 125.39 1.3 nd 9.91 0.07 143.58

S. androgynus nd nd 4.50 0.22 nd 138.14 5.81 142.64
M. pterygosperma nd 1.32 0.04 95.84 1.98 nd 20.79 0.35 117.95
P. pinnata nd nd 28.48 1.88 nd 23.71 1.38 52.19
C. caudatus nd nd 51.28 4.06 nd 0.90 0.05 52.19
H. sibthorpioides 1.57 0.01 nd 37.51 0.66 nd 10.85 0.16 49.93
V. unguiculata nd nd 27.35 0.75 12.97 0.3 3.33 0.11 43.65
C. papaya nd nd 18.85 0.11 11.95 0.28 5.47 0.22 36.27
Se. edule 12.49 0.14 nd 13.81 0.17 nd 9.72 0 .15 36.03
M. citrifolia nd nd 23.67 1.62 nd 9.75 0.69 33.42
P. scutellaria nd nd 12.67 0.12 6.87 0.33 12.95 0.26 32.49
S. grandiflora nd nd 2.76 0.05 nd 18.47 0.31 21.23
C. asiatica 0.13 0.00 nd 12.31 0.41 nd 8.57 0.38 21.00
A. schoenoprasum 2.69 0.02 nd 4.46 0.08 nd 7.65 0.15 14.79
A. irregularis nd nd 7.42 0.06 nd 2.10 0.01 9.52
O. americanum nd 2.12 0.05 1.89 0.10 0.74 0.04 2.47 0.18 7.22
P. indica 0.90 0.03 nd 5.21 0.26 nd 0.28 0.02 6.39
N. scutellarius nd nd 3.69 0.09 nd 1.74 0.07 5.43
T. triangulare nd nd 0.41 0.03 nd 3.52 0.16 3.93
S. torvum fruits 2.30 0.06 nd 0.66 0.01 nd nd 2.96
P. melastomoides nd 0.33 0.02 1.76 0.20 nd 0.25 0.03 2.34
E. elatior nd nd 1.18 0.06 nd nd 1.18
Sa. edule nd nd 0.44 0.01 nd nd 0.44
P. oleracea nd nd 0.30 0.02 nd nd 0.3
nd, not detected above the limit of detection (3.9 lg myricetin, 5.6 lg luteolin, 22 lg apigenin, 4.7 lg kaempferol per g dry weight basis).
 JOURNAL OF FUNCTIONAL FOODS 4 ( 20 1 2) 3 3 934 7 343

leaves had significant myricetin content with 12.49 and
8.28 mg/100 g fw, respectively. Although flavones were not as
prevalent as flavonols in vegetables, V. unguiculata (L.) Walp
leaves and C. papaya L. flowers had 13 and 12 mg apigenin/
100 g, respectively. To the best of our knowledge, this is the first
report of flavonoid content in the edible parts of A. irregularis
(C.Presl) Ching (9.52 mg/100 g), Polyscias scutellaria (Burm.f.)
Fosb. (32.49 mg/100 g), Sa. edule Hassk (0.44 mg/100 g), and S.
grandiflora (L.) Pers. (21.23 mg/100 g).
The flavonoid content of vegetables in the present study
was similar to those measured previously (Andarwulan
et al., 2010). However, Centelia asiatica (L.) Urb. and Pluchea
indica (L.) Less had more than 60-fold higher myricetin con-
tent compared to our previous study. While the reasons for
these differences are unclear, environmental, seasonal, and
soil variation may impact leafy green vegetable polyphenol
content (Bano et al., 2004; Reimberg, Renata, & Yariwake,
2009).
We quantified common vegetable flavonoids as aglycone
equivalents following their hydrolysis. This approach may
underestimate flavonoid content, due to the diversity of flavo-
noids in plants. For example, S. torvum fruits also contain iso-
flavones (Arthan et al., 2002) and A. occidentale leaves also
contain robustaflavone, agathisflavone, and amentoflavone
(Arya, Babu, Ilyas, & Nasim, 1989). In addition, it should be
noted that plant flavonoids are mainly conjugated to sugars.
For example, glycosylated or rhamnosyl flavonoids were pre-
viously identified in M. citrifolia leaves (Deng, West, & Jensen,
2008) and C- and O-glycosyl flavones were identified in Se.
edule (350 mg/100 g dw) (Siciliano, De Tommasi, Morelli, &
Braca, 2004). Nonetheless, this work is an important first step
in characterizing the flavonoids in a number of underutilized
medicinal vegetables.
Chlorogenic acid, caffeic acid, and ferulic acid were pres-
ent at 0.0452.53 mg/100 g fw in vegetables (Table 3). P. pinnata
leaves had the most phenolic acids among the vegetables
studied, with a distribution of 90% chlorogenic acid and 10%
ferulic acid. All vegetables contained phenolic acids, although
T. triangulare leaves, A. schoenoprasum, and S. grandiflora
flowers had less than 1 mg/100 g. M. citrifolia, A. occidentale,
P. indica, and P. pinnata leaves did not have caffeic acid above
the LOD. A. irregularis leaves and Hydrocotyle sibthorpioides
Lmk. consisted only of chlorogenic and caffeic acid, while
S. grandiflora flowers had solely ferulic acid. To the best of
our knowledge, this is the first report of phenolic acid content
in P. pinnata, M. pterygosperma Gaertn., A. irregularis (C.Presl)
Ching, N. scutellarius (Burm.f.) Merr., Sa. edule Hassk, Se. edule
(Jacq.) Swartz, and S. grandiflora (L.) Pers.
Chlorogenic acid was the most abundant phenolic acid in 18
of the 24 vegetables. Caffeic acid was the second most abun-
dant phenolic acid among the vegetables studied. Caffeic acid
was most enriched in P. indica and C. caudatus leaves with 8.65
and 3.64 mg/100 g fw, respectively. P. pinnata, M. pterygosperma,
and C. caudatus leaves had the greatest ferulic acid content
among vegetables, with 35 mg/100 g fw.
It should be noted that phenolic acids besides those quan-
tified in the present study could significantly contribute to
vegetable phenolic acid content. For example, gallic, proto-
catechuic, p-hydroxybenzoic, cinnamic, and p-coumaric acids
are also found in A. occidentale leaves (Kogel & Zech, 1985).

Table 3 Specific phenolic acids and sum phenolic acids in 24 medicinally used Indonesian vegetables.

Vegetable Phenolic acids (mg/100 g fw)

Chlorogenic acid Caffeic acid Ferulic acid Total

P. pinnata 47.02 0.81 nd 5.02 0.16 52.53 0.97

S. torvum 33.14 1.63 2.56 0.08 0.32 0.01 35.76 1.71
P. indica 20.00 0.24 8.65 0.46 nd 28.48 0.67
H. sibthorpioides 24.27 0.51 1.35 0.03 nd 25.51 0.54
P. melastomoides 17.47 0.39 1.11 0.01 0.17 0.00 18.57 0.40
A. occidentale 13.53 0.38 nd 2.88 0.12 16.70 0.32
P. scutellaria 14.13 0.41 1.69 0.03 0.80 0.04 16.30 0.47
E. elatior 14.06 1.11 0.96 0.03 0.13 0.00 15.01 1.15
M. pterygosperma 6.65 0.16 2.93 0.08 4.41 0.16 13.50 0.40
C. asiatica 9.22 0.34 1.19 0.02 1.81 0.09 11.86 0.44
C. caudatus leaves 4.54 0.18 3.64 0.14 3.14 0.28 10.92 0.44
V. unguiculata 4.26 0.12 2.02 0.04 1.38 0.09 7.34 0.24
P. oleracea 5.79 0.07 0.55 0.00 0.22 0.01 6.32 0.06
Se. edule 5.80 0.12 0.55 0.00 0.13 0.00 6.20 0.12
Sa. edule 4.17 0.13 1.05 0.02 0.16 0.00 5.13 0.15
S. androgynus 3.38 0.32 1.13 0.02 1.10 0.04 5.12 0.29
M. citrifolia 2.31 0.05 nd 0.76 0.04 3.24 0.09
A. irregularis 2.58 0.16 0.47 0.01 nd 2.88 0.16
C. papaya 0.77 0.00 1.03 0.00 0.75 0.02 2.30 0.02
O. americanum 0.32 0.01 2.03 0.06 0.16 0.00 2.23 0.06
N. scutellarius 0.86 0.02 1.15 0.01 0.24 0.00 1.82 0.03
T. triangulare 0.38 0.01 0.41 0.00 0.09 0.00 0.68 0.01
A. schoenoprasum 0.08 0.01 0.36 0.01 0.10 0.00 0.35 0.01
S. grandiflora nd nd 0.04 0.00 0.04 0.00
nd, not detected above limit of detection (1.6 lg ferulic acid/g dry weight, 1.9 lg chlorogenic acid/dry weight, and 1.6 lg caffeic acid/g dry
weight).
344 JOURNAL OF FUNCTIONAL FOODS 4 ( 2 0 1 2 ) 3 3 9 3 4 7

S. torvum fruits also have methyl caffeate (Takahashi et al.,
2010).
Chlorogenic acid is present in a number of vegetables
including lettuce (3.78 mg/100 g fw), broccoli (1.78 mg/100 g
fw), and endive (101 mg/100 g fw) (Neveu et al., 2010). Caffeic
acid is present in fresh herbs such as rosemary, sage, thyme,
and oregeno from 2.08 to 11.7 mg/100 g (Neveu et al., 2010).
Ferulic acid has previously been identified in vegetables from
0.21 to 3.01 mg/100 g fw, and is also found in beans and dried
herbs (Neveu et al., 2010). Thus, consumption of P. pinnata,
M.pterygosperma, and C. caudatus may increase dietary chloro-
genic acid and ferulic acid from vegetables. Animal studies
suggest that chlorogenic acid and ferulic acid improve antiox-
idant status and increase phase 2 detoxification enzymes.
Dietary chlorogenic acid improved hepatic glutathione redox
status in rats subjected to azoxymethane-induced colon can-
cer (Park, Davis, Liang, Rosenberg, & Bruno, 2010). Ferulic acid
supplementation increased intestinal glutathione expression
of glutathione-S-transferase and quinone reductase in rats
(Bolling et al., 2011).
Total content of phenols of vegetables ranged from 21.01 to
847.41 mg GAE/100 g (Table 4). A. occidentale and P. indica leaves
had more than twofold higher total phenols content than
other vegetables. A. occidentale leaves also had the greatest fla-
vonoid content and the 6th highest phenolic acid content.
The comparatively higher total phenols content suggests that
flavones or other polyphenols are abundant in A. occidentale
leaves (Siciliano et al., 2004). Similarly, P. indica flavonoid, phe-
nolic acid, and anthocyanin contents do not appear to ac-
count for its high total phenols content. Other non-phenolic
reducing agents present in the ethanol extracts of vegetables,
such as ascorbic acid, are known to contribute to total content
of phenols. However, ascorbic acid was not correlated with
total phenols of vegetables (Fig. 2). More work is needed to
define the constituents in A. occidentale and P. indica leaves
that contribute to their increased total content of phenols.
The vegetables in the present study did not have particu-
larly abundant anthocyanin content, ranging from 0.07 to
4.44 mg/100 g fw (Table 4). M. pterygosperma leaves, S. torvum
Swartz fruits, and E. elatior flowers had 34 mg/100 g, which
is considerably less than the 26589 mg/100 g fw found in ber-
ries (Neveu et al., 2010). Nevertheless, the anthocyanin con-
tent illustrates the diversity of polyphenolic compounds,
and may contribute synergistically or additively to the medic-
inal properties of these vegetables.
Leafy green carotenoids are composed of b-carotene, lutein,
neoxanthin, zeaxanthin, and violaxanthin, with varying bioac-
cessibility (Chandrika, Basnayake, Athukorala, Colombagama,
& Goonetilleke, 2010). It is estimated that 85% of total vitamin A
activity from leafy green carotenoids is from b-carotene (Ball,
2000). The b-carotene content of vegetables in the present
study ranged from 0.13 to 2.25 mg/100 g. For comparison, ice-
berg lettuce (0.30 mg/100 g), green leaf lettuce (4.4 mg/100 g),
and spinach (5.6 mg/100 g) have equivalent or more b-carotene
than the vegetables in the present study (USDA-ARS., 2011). The
total carotenoid content of vegetables analyzed was 0.36
13.96 mg b-carotene equivalents (bCE)/100 g fw (Table 4). M.
pterygosperma leaves had the most carotenoids with 13.96 mg
b-carotene equivalents (bCE)/100 g and 2.25 mg b-carotene/
100 g. C. caudatus and P. indica leaves also had relatively higher
carotenoid content compared to other vegetables, with 9.55
and 8.74 mg bCE/100 g FW. The ratio of b-carotene to total

Table 4 Total phenols, anthocyanin, and carotenoids in 24 medicinally used Indonesian vegetables.
Vegetable Total phenols Total anthocyanin b-Carotene Total carotenoids Ascorbic acid
(mg GAE/100 g fw) (mg/100 g fw) (mg/100 g fw) (mg bCE/100 g fw) (mg/100 g fw)

A. occidentale 847.41 15.05 0.37 0.02 0.73 0.06 5.42 0.36 494.43 0.94
P. indica 742.54 24.78 0.27 0.01 1.70 0.05 8.74 0.34 15.44 0.00
C. caudatus 342.06 0.37 0.78 0.05 1.35 0.03 9.55 0.27 108.83 0.50
E. elatior 256.99 0.98 4.42 0.11 nd 0.83 0.05 16.90 0.00
C. asiatica 200.52 2.05 1.08 0.05 1.16 0.12 5.95 0.38 24.04 0.01
P. pinnata 189.08 0.32 0.41 0.01 0.51 0.10 3.29 0.27 12.03 0.19
P. scutellaria 179.88 2.00 1.64 0.08 0.85 0.11 7.64 0.57 15.61 0.00
S. torvum 158.92 0.94 4.44 0.14 0.13 0.01 0.87 0.03 54.78 0.04
H. sibthorpioides 144.81 1.14 0.77 0.03 0.24 0.01 2.75 0.32 21.85 0.00
S. androgynus 138.01 0.47 1.53 0.11 1.63 0.02 5.15 0.07 190.83 0.82
P. melastomoides 121.51 1.64 0.75 0.01 1.48 0.16 5.12 0.20 45.13 0.00
V. unguiculata 112.55 2.20 1.23 0.08 0.38 0.07 3.31 0.09 58.87 0.84
M. pterygosperma 107.00 2.87 3.25 0.01 2.25 0.05 13.96 0.19 90.28 0.85
Sa. edule 87.65 3.43 2.38 0.13 0.02 0.00 1.29 0.08 29.16 0.00
O. americanum 86.89 0.70 0.11 0.01 1.56 0.20 7.35 0.45 146.37 0.17
P. oleracea 82.66 1.12 0.24 0.01 0.94 0.08 5.48 0.22 21.48 0.43
M. citrifolia 72.72 0.38 1.12 0.04 0.33 0.00 3.28 0.21 30.71 0.00
C. papaya 66.75 0.20 1.33 0.08 0.13 0.01 0.84 0.06 117.15 0.01
Se. edule 66.46 0.71 0.78 0.04 1.77 0.23 2.83 0.03 31.74 0.42
T. triangulare 64.64 0.50 0.23 0.01 0.97 0.01 4.22 0.14 25.52 0.01
A. irregularis 61.56 1.25 0.07 0.00 0.89 0.07 6.15 0.08 69.85 0.99
N. scutellarius 40.36 0.09 1.42 0.07 0.21 0.02 2.69 0.01 34.30 0.13
S. grandiflora 38.43 1.53 0.22 0.01 0.01 0.00 0.36 0.03 33.52 0.03
A. schoenoprasum 21.01 + 0.13 0.46 0.01 0.08 0.00 0.65 0.06 24.59 0.02
nd, not detected above limit of detection; GAE, gallic acid equivalents; bCE, b-carotene equivalents.
 JOURNAL OF FUNCTIONAL FOODS 4 ( 20 1 2) 3 3 934 7 345

Fig. 2 (A) Principal component scores and (B) loading plot of Principal Component Analysis of functional components of
vegetables of Indonesian origin.

carotenoids was greatest in Se. edule (63%), S. androgynus (32%), components explaining 33% and 24% of the data variation
and P. melastomoides leaves (29%). Minimal to no b-carotene was (Fig. 2). The loading plot illustrates the relationships among
detected in S. grandiflora (2%), Se. edule (3%), and E. elatior flowers variables, where two vectors with an angle less than 90 are
(none detected). Thus, a substantial amount of other carotenes positively correlated and two vectors with an obtuse angle
or xanthophylls may account for the total carotenoid values for (>90) are negatively correlated (Sartono, Affendi, Syahfitri,
these vegetables. Sumertajaya, & Anggraeni, 2003). Total phenols was positively
Many of the vegetables sampled contained significant quan- correlated with the flavonoid and phenolic acid content.
tities of ascorbic acid. Ascorbic acid content ranged from 12.03 Ascorbic acid was more strongly correlated with flavonoid
to 494.43 mg/100 g fw (Table 4). A. occidentale leaves had the and total phenol content than total carotenoids or phenolic
most ascorbic acid with 494.43 mg/100 g, while S. androgynus acids. Carotenoid and b-carotene content was not well-corre-
leaves. A 30 g serving (1 c) of A. occidentale leaves would lated with polyphenol content. A. occidentale L. (jambu mete)
provide twice the ascorbic acid of an orange (USDA-ARS, was distinguished by its high ascorbic acid, flavonoids, and
2011). O. americanum, C. caudatus, and C. papaya leaves had more
than 100 mg/100 g FW. The majority of Indonesian vegetables
had ascorbic acid content greater than typical western vegeta-
bles such as spinach (28 mg/100 g), green leaf lettuce (9.2 mg/ Table 5 Moisture and protein of underutilized medicinal
100 g), and iceberg lettuce (2.8 mg/100 g) (USDA-ARS, 2011). vegetables of Indonesian origin.
Carotenoid and anthocyanin pigmentation is dependent Vegetable Moisture Protein
upon protein denaturation following heat processing of cer- (g/100 g fw) (g/100 g fw)
tain vegetables, such as sweet potatoes. While carotene and
A. schoenoprasum 92.30 0.32 0.43 0.03
protein content were not correlated in tubers, this relation- T. triangulare 91.83 0.00 1.79 0.04
ship in other vegetables is unclear (Novita, Hartana, & Soehar- S. grandiflora 90.23 0.09 2.41 0,01
sono, 1996). The vegetables analyzed had 0.48 g protein/100 g E. elatior 89.77 0.21 0.96 0.05
fw (Table 5). The protein content among vegetables analyzed A. irregularis 89.27 0.89 3.61 0.09
varied more than 10-fold, and are comparable to spinach C. papaya 88.91 0.45 3.50 0.02
Sa. edule 88.39 0.10 2.69 0.03
and lettuce contents (USDA-ARS, 2011). Protein content in
P. oleracea 88.07 0.35 2.36 0.01
vegetables was not correlated with polyphenol, carotenoid,
P. melastomoides 87.68 0.01 3.33 0.21
or ascorbic acid content. O. americanum 87.42 0.47 4.56 0.07
The moisture content of vegetables was 7592%. Thus, on Se. edule 86.68 0.48 3.33 0.03
a dry weight basis, some vegetables may comparatively be M. citrifolia 85.46 0.06 2.50 0.02
better sources of functional constituents than others. This P. pinnata 85.43 1.57 3.08 0.03
is a consideration to utilizing these vegetables for ingredients V. unguiculata 84.36 0.38 5.95 0.03
H. sibthorpioides 84.30 0.11 3.01 0.05
or supplements. On a dry weight basis, A. occidentale leaves
P. scutellaria 82.31 0.18 2.44 0.06
and P. indica leaves had the highest total phenols with 4.4
N. scutellarius 82.28 0.13 4.32 0.04
and 3.9 g GAE/100 g. S. androgynus leaves was the best source C. asiatica 81.72 0.30 0.79 0.03
of flavonoids with 831 mg/100 g. S. torvum fruits and P. pinnata A. occidentale 80.82 0.09 6.96 0.16
leaves were the best sources of phenolic acids, with 179 and P. indica 80.81 1.50 4.49 0.10
357 mg/100 g. C. caudatus 80.3 2.53 4.22 0.12
PCA was used to examine the relationships among the S. torvum 79.89 1.22 3.11 0.06
S. androgynus 78.19 0.49 8.31 0.11
functional components of vegetables. Principal component
M. pterygosperma 75.27 0.14 7.76 0.11
scores were able to distinguish vegetables, with the first two
346 JOURNAL OF FUNCTIONAL FOODS
total phenols values. E. elatior (Jack) R.M.Sm. (kecombrang)
was distinguished from other vegetables by its relatively
higher anthocyanin and phenolic acid content. O. americanum
L. (kemangi) were distinguished by the combination of
increased ascorbic acid and carotenoids. The unique distribu-
tions of functional components in these plants make them
intriguing candidates for further research into their tradi-
tional medicinal uses.
4. Conclusions
In conclusion, underutilized vegetables of Indonesian origin
were rich sources of polyphenols, ascorbic acid, and carote-
noids to varying degrees. This work provides a basis for fur-
ther characterization of polyphenols, carotenoids and other
constituents of these vegetables. A. occidentale, S. androgynus,
M. pterygosperma, and P. indica were rich in polyphenol content
and are promising candidates to further investigate for func-
tional uses. More work is needed to determine the safety of
increased consumption of these plants, particularly outside
the context of their traditional uses and intakes. Further stud-
ies are necessary to establish the efficacy and plausible mech-
anisms for the medicinal uses of these vegetables.
Acknowledgement
This work was supported by the Southeast Asian Food and
Agricultural Science and Technology (SEAFAST) Center.
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at doi:10.1016/j.jff.2012.01.003.
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