Food Chemistry xxx (2016) xxxxxx

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Nutrients and natural toxic substances in commonly consumed

Jerusalem artichoke (Helianthus tuberosus L.) tuber
Kunchit Judprasong a,, Nidthida Archeepsudcharit a, Kedsiri Chantapiriyapoon b,
Pharrunrat Tanaviyutpakdee a, Piya Temviriyanukul a
a
Institute of Nutrition, Mahidol University, Putthamonthon 4 Rd., Salaya, Phutthamonthon, Nakhon Pathom 73170, Thailand
b
Office of Agricultural Research and Development, Region 6, Tapoon, Klung, Chanthaburi 22110, Thailand

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

Article history: This study determined nutrients, chemical contaminants, (insecticide residues and heavy metals), and
Received 27 June 2016 natural toxic substances (nitrate, nitrite, cyanide, oxalate, phytate, and trypsin inhibitor) in tubers of
Received in revised form 5 September 2016 Jerusalem artichokesKaentawan in the Thai languagegrown in four major provinces in Thailand.
Accepted 17 September 2016
They were purchased, prepared, homogenized, and freeze-dried for further analysis using standard meth-
Available online xxxx
ods. All Kaentawan samples contained considerable amounts of fructans and dietary fiber
(15.40.2 g and 3.20.8 g/100 g fresh weight [FW], respectively), as well as potassium and iron
Keywords:
(33961 and 0.320.05 mg/100 g FW, respectively). All samples had very low amounts of insecticide resi-
Jerusalem artichoke
Kaentawan
dues (37 compounds), cyanide, and trypsin inhibitor, as well as Pb, Cd, nitrate, and nitrite (0.820.09,
Nutrient 0.100.02, 1.917.5, and 0.010.24 mg kg 1 FW, respectively), in addition to oxalate and phytate
Toxic substance (149 and 0.170.02 mg/100 g FW, respectively). This studys data can be used for food composition data-
Contaminant bases and for safety consumption.
2016 Elsevier Ltd. All rights reserved.

1. Introduction
Kaentawan, or Jerusalem artichoke (Helianthus tuberosus L.), is
known as an easy growing crop plant (Bach, Clausen, &
Edelenbos, 2015; Kays & Nottingham, 2008). Kaentawan tuber
has been reported as a rich source of inulin and fructo-
oligosaccharide (FOS). Many studies have shown that inulin-type
fructans have beneficial effects in terms of dietary fiber and prebi-
otics. Since Kaentawan has a low caloric value, is fat-free, and has a
high mineral content (Kays & Nottingham, 2008), it can be devel-
oped and applied as a functional food. Generally, this tuber, though
a good source of nutrients, can also contain toxic substances in
terms of either chemical contaminants (i.e., insecticide residues
or heavy metals) or natural toxins (i.e., nitrate, nitrite, cyanide,
phytate, oxalate, and trypsin inhibitor) (Jaworska, 2005; Kays &
Nottingham, 2008).
For chemical contaminants, insecticides contain a variety of
toxic substances. Most organochlorine (OC) insecticides have been
banned, since they can persist in the environment. Carbamate
(CRB) and organophosphorus (OP) can replace OC which is most
widely used for crop plants (Bai, Zhou, & Wang, 2006). Pyrethroid
Corresponding author.
E-mail address: Kunchit.jud@mahidol.ac.th (K. Judprasong).
(PYR) is another choice and has naturally synthetic analogues. PYR
insecticides have greater photostability and relatively low toxicity
compared to OC, OP, and CRB insecticides (Ware & Whitacre,
2004). Heavy metal contents can contaminate the environment
before plants are grown, such as in water and soil, (McGrath,
Zhao, & Lombi, 2001). For example, cadmium (Cd), lead (Pb),
arsenic (As), and mercury (Hg) may be found at certain levels in
plants (Onianwa, Adetola, Iwegbue, Ojo, & Tella, 1999).
For natural toxins, nitrate and nitrite are naturally present in a
wide range of vegetables. They accumulate in the stem, leaves, and
roots of plants and may relate to high levels of ammonia or nitrates
in the soil (Nicholson, 2007). Oxalate is widely found in plants in a
water soluble form (sodium, potassium, and ammonium) and an
insoluble (calcium) form, which may be present at different levels
in different plant parts (Holloway, Argall, Jealous, Lee, & Bradbury,
1989; Judprasong, Charoenkiatkul, Sungpuag, Vasanachitt, &
Nakjamanong, 2006). Oxalate can bind strongly with calcium, iron,
and magnesium (Noonan & Savage, 1999), which can have adverse
health effects (Jaworska, 2005). A diet high in oxalates can cause
excessive urinary excretion of oxalate (hyperoxaluria) and can
increase the risk of developing kidney stones (Nguy n & Savage,
2013). Trypsin is a serine protease that is produced in the pancreas
as zymogen trypsinogen and stored in active form (Horton, Moram,
Acrimgeour, Perry, & Rawn, 2006). Trypsin inhibitor (TI) inhibits

http://dx.doi.org/10.1016/j.foodchem.2016.09.116
0308-8146/ 2016 Elsevier Ltd. All rights reserved.

Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
2 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx
the activities of protein digesting enzymes in the digestive tract
and reduces the bodys capability to use protein in foods
(Srinivasan, Giri, Harsullkar, Gatehouse, & Gupta, 2005).
Currently, there is limited, to no, information available on nutri-
ents, chemical contaminants (insecticide residues and heavy met-
als), and natural toxic substances (nitrate, nitrite, cyanide, oxalate,
phytate, and trypsin inhibitor) in Kaentawan tubers. Consequently,
these components should be investigated to ensure healthy and
safe consumption.
2. Materials and methods
2.1. Samples and sample preparation
Kaentawan tubers were purchased from four major growing
areas in Thailand (Nakhon Pathom, Khonkaen, Nakhon Ratchasima,
and Phetchabun provinces) during April to May 2014. The tubers
were transported to the laboratory at Institute of Nutrition, Mahi-
dol University. Kaentawan tubers from each source were classified
to two groups, namely, fresh Kaentawan without skin and boiled
Kaentawan without skin. Each sample was peeled, weighed edible
part, and washed with deionized water (DI water). Each sample
was homogenized using a food processor (Mara, Thailand) and
separated into two portions as fresh sample and dried sample by
freeze drying (lyophilization). The prepared samples were then
stored at 20 C until analysis.
2.2. Reagents and standards
Deionized water was obtained by means of a Millipore water
purification system with resistivity 18.2 MX cm 1 (Millipore
RiOs-DITM134, Bedford, MA, USA). Nitric acid, perchloric acid, sul-
furic acid, hydrochloric acid, acetic acid, cadmium standard, lead
standard, silver nitrate, potassium cyanide, potassium hydroxide,
sodium hydroxide, sodium chloride, oxalic acid dehydrate (99.5
102.0%), phytic acid, sodium salt hydrate, hexane, and other chem-
icals were obtained from MERCK (Germany). Acetone, dichloro-
methane, ethyl acetate, 2, 2, 4 trimethylpentane (Isooctane),
silica gel (60200 mesh), and sodium sulfate anhydrous (1260
mesh) were analytical grade from Sigma-Aldrich, USA. All other
chemicals and solvents used were of analytical and HPLC grade.
2.3. Nutrient determination
Nutrient analyses (proximate compositions and minerals) were
conducted using standard AOAC methods (AOAC, 2012). All sam-
ples were analyzed by the accredited ISO 17025:2005 laboratory
at the Institute of Nutrition, Mahidol University, Thailand. The
results of all measurements were presented as mean standard
deviation in fresh weight of the edible portion.
2.3.1. Proximate composition
Total nitrogen was determined using the Kjeldahl method fol-
lowing AOAC method number 981.10 (AOAC, 2012) and calculated
into protein content using 6.25 as the specific (Jones) factor. Mois-
ture was analyzed using a drying method at 100 2 C until a con-
stant weight, following AOAC method no. 952.45 (AOAC, 2012).
Crude fat was determined by acid digestion prior to continuous
extraction using petroleum ether in Soxtec system following AOAC
method no. 945.16 (AOAC, 2012). Ash was analyzed by incinerating
all organic matter at 550 5 C in accordance with AOAC method
no. 945.46 (AOAC, 2012). Carbohydrate was calculated using the
formula 100-moisture-protein-fat-ash; energy was calculated by
Atwater factor (4 for protein and carbohydrate and 9 for total
Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
fat). Total dietary fiber was determined using enzymatic gravimet-
ric method following AOAC method no. 985.29 (AOAC, 2012).
2.3.2. Fructans
Fructans (inulin and fructooligosaccharides) were analyzed fol-
lowing the AOAC method no. 997.08 (AOAC, 2012) by extracting
with hot water and hydrolysing by inulinase (SigmaAldrich,
USA, I2017 inulinase from Aspergillus niger, CAS Number 9025-
67-6 which has enzyme activity of 1740 inu/g). Both hot water
extracted and enzyme hydrolysed fractions were derivatized into
volatile oxime-trimethylsilyl derivatives. Each individual sugar,
both before and after enzyme hydrolysis, was then determined
by high temperature gas chromatography (Joye & Hoebregs,
2000; Judprasong, Tanjor, Puwastien, & Sungpuag, 2011) and the
fructans in each sample were calculated.
2.3.3. Minerals
The acid solution dissolved from ash residue after incineration
at 550 C for 2 h was used for calcium, sodium, and potassium con-
tent analyses by flame atomic absorption spectrophotometer (AAS)
following method no. 975.03 (AOAC, 2012). The acid solution was
also determined for phosphorus by the gravimetric method follow-
ing AOAC no, 920.55 (AOAC, 2012). Acid digestion in a closed
Teflon vessel was employed for determining magnesium, iron, cop-
per, and zinc using an inductively coupled plasma optical emission
spectrophotometer (ICP-OES), method no. 984.27 (AOAC, 2012).
2.4. Toxic substances determination
2.4.1. Insecticide residue
Each sample (about 20 g) was weighed into an Erlenmeyer flask
containing 20 g of DI water. Fifty mL acetone, 50 mL dichloro-
methane, and a little sodium chloride were added to the mixture.
The extraction was blended and mixed with ultra-turrax at high
speed for 3 min. The organic phase was transferred into a beaker
and dried with Na2SO4 for 10 min. For the organic phase, 50 mL
was taken and reduced in volume to 35 mL with rotary evapora-
tor. Thereafter, it was adjusted to the volume of 5 mL with ethyl
acetate and re-concentrated (repeat the evaporation twice with
ethyl acetate to make sure the complete removal of acetone). The
solution was injected into gas liquid chromatography, GLC (Agilent
6890 N, USA), for organophosphorus analysis. Another 2 mL of
solution was cleaned up by silica gel and then eluted with hex-
ane:dichrolomethane (4:1) and injected into GLC (Agilent 6890 N,
USA) for organochlorine and pyrethroids analyses. The solution
was dissolved into methanol:water and injected into high perfor-
mance liquid chromatography, HPLC (Agilent 1100 Series, USA),
for carbamate analysis.
2.4.2. Heavy metals
Each sample (0.5 g) was extracted with 5 mL of 65% HNO3 and
1 mL HClO4 into a Teflon jar. The extraction was digested at
100 C for 9 h in a hot air oven (Memmert, USA) and then left to
cool under the fume hood. The solution was transferred to a
25 mL volumetric flask and diluted with DI water to the mark.
Heavy metals (Pb and Cd) were determined using Inductively Cou-
ple Plasma-Optical Emission Spectrometer (ICP-OES, PerkinElmer,
Optima 4300, USA) with WinLab32 software against standard addi-
tion methods.
2.4.3. Nitrite and nitrate
Nitrate and nitrite content in all study samples were deter-
mined by the cadmium column method (AOAC, 2012). Each sample
(5 g) was extracted with 5 mL saturated borax solution and 100 mL
hot water, mixed thoroughly, heated in a boiling water bath at
100 C for 20 min, and then shaken repeatedly. The solution was
 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx
diluted with DI water, shaken, and left to stand at room tempera-
ture for 30 min. Two grams of activated charcoal was added to the
solution to remove pigment and filtered through Whatman No.1
filter paper.
2.4.3.1. Determination of nitrite content. A 3 mL filtrate was pipetted
into a test tube along with 3 mL of color mixture reagent. The solu-
tion was mixed, left to stand for 15 min at room temperature in the
dark, and then measured by using a spectrophotometer (Shimadzu
UV-1601PC UVVisible, Japan) at 540 nm.
2.4.3.2. Reduction of nitrate to nitrite. Twenty mL of the filtrate was
pipetted into the reservoir on the top of the column; thereafter,
5 mL of ammonium buffer solution pH 9.69.7 and 80 mL of DI
water were added. The effluent was collected from the column in
a 100 mL volumetric flask (eluate). Three mL of the eluate was
pipetted into a test tube for determining nitrite in the eluate. Three
mL of color reagents were added to the eluate. The solution was
vortex-mixed, left 15 min at room temperature in the dark, and
measured by using a spectrophotometer at 540 nm.
2.4.4. Cyanide
Cyanide in all samples was determined using titration by fol-
lowing AOAC official method number 920.30 (AOAC, 2012). Each
sample (1.0 g) was extracted with DI water, a small amount of
PbCO3, and diluted to 100 mL with DI water. The extraction was
mixed thoroughly and filtered through a dry filter. The solution
(25 mL) was transferred into an Erlenmeyer flask, along with the
addition of 100 mL DI water, 5 mL NaOH solution, and 2 g of KI.
Each sample was then titrated to faint opalescence with AgNO3
solution. The calculation of cyanide content was made.
2.4.5. Oxalate
Oxalate content in all samples was determined by the HPLC
method (Judprasong et al., 2006; Savage, Vanhanen, Mason, &
Ross, 2000). In brief, each sample (1 g) was extracted with 50 mL
HCl and heated in a water bath at 80 C for 15 min. The extraction
was adjusted to volume to 50 mL with HCl, centrifuged for 15 min
at 3000 rpm, and filtered through a 0.45 lm cellulose acetate
membrane. The solution was injected into the HPLC system with
a UV detector set at 210 nm. The organic acid, including oxalic acid,
was separated in an HPLC column (Aminex HPX-87H,
300 mm  7.8 mm), using an isocratic elution at 0.65 mL/min with
0.00125 M sulfuric acid as a mobile phase. The amount of oxalic
acid in each sample was determined against a standard calibration
curve of oxalic acid and shown as mg oxalate in 100 g sample.
2.4.6. Phytate
Phytate contents in all samples were determined using spec-
trophotometer following the method of Gao et al. (2007). Each
sample (0.5 g) was extracted with 10 mL of 2.4% HCl in a 14 mL
screw-cap tube for 16 h. The extraction was centrifuged at
1000 g under 10 C for 20 min. The crude acid extracts were trans-
ferred to screw-cap tubes containing 1 g of NaCl and shaken at
350 rpm for 20 min. Thereafter, they were settled at 4 C for
60 min (or 20 C for 20 min) and centrifuged 1000 g under
10 C for 20 min. The clear supernatants were diluted 25 times in
screw-cap tubes with 24 mL of DI water. Three mL of the diluted
supernatants were pipetted into test tubes, with the addition of
1 mL of wade reagent, and mixed on vortex. Finally, they were cen-
trifuged at 1000 g under 10 C for 10 min and then measured for
absorbance using a spectrophotometer (Shimadzu UV-1601PC
UVVisible, Japan) at 500 nm.
Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
3
2.4.7. Trypsin inhibitor
Trypsin inhibitor activity in all samples was determined accord-
ing to AOCS. (1997). Each sample (1 g) was extracted with 50 mL of
0.01 N NaOH solution. The extraction was slowly agitated at room
temperature for 3 h, adjusted for pH to 8.410.0, and diluted with
DI water (6080% of standard). DI water was added to the solutions
(0, 0.6, 1.0, 1.4, and 1.8 mL) to bring the total volume up to 2 mL in
each test tube. Two mL of trypsin solution was added to each test
tube, mixed on vortex, and immediately placed in a 37 C water
bath. Time was prepared for a 10 min interval. Thereafter, 5 mL
of BAPA solution was added, mixed on vortex, and incubated in a
37 C water bath; timing of the 10 min interval started immedi-
ately. After the 10 min interval, the reaction in each tube was
stopped by adding 1 mL of acetic acid solution and mixed on vor-
tex. The solution was centrifuged at 10,000 rpm for 5 min. Clear
supernatant was determined for absorbance in a spectrophotome-
ter at 410 nm.
3. Results and discussion
Nutrients and toxic substances were analyzed by the accredited
ISO 17025:2005 laboratory in which a quality control system had
been regularly undertaken. For some important quality parame-
ters, most of these were studied along with standard reference
materials or spiked standard (% recovery) as shown in Tables 1A
and B. For precision, in-house quality control (QC) samples were
used, for instance, milk powder (for total nitrogen, moisture, and
fat analyses) and defatted soybean flour (for ash, total dietary fiber,
and mineral analyses). Each analyzed parameter was conducted in
duplicate or triplicate with each accepted criteria.
3.1. Nutrients
Since Kaentawan tubers have very thin skin, the percentage of
edible portion in this plant was 82.5 1.2% (Table 2), which is close
to what Judprasong et al. (2011) have reported (79.0%).
3.1.1. Proximate compositions
As shown in Table 2, fresh and boiled Kaentawan tuber con-
tained high moisture (7880 g/100 g fresh weight [FW]), moderate
protein (2.22.3 g/100 g FW), and very low fat (0.1 g/100 g FW).
Carbohydrate content was the second main nutrient (17.5
18.6 g/100 g FW). However, most carbohydrate was in the form
of fructans (15.416.9 g/100 g FW) and dietary fiber (3.2
4.1 g/100 g FW). Fructan levels in this study were slightly lower
than previously reported by Judprasong et al. (2011), but were in
the range of 14.120.4 g/100 g FW as reported by Tanjor,
Judprasong, Chaito, and Jogloy (2012). Total sugar (1.1
1.5 g/100 g FW), mainly sucrose, was found at the same level as
in a previous study (Judprasong et al., 2011).
Table 1A
Accuracy test using standard reference material (SRM) for nutrient substances (per
100 g).
Analytical components Certified value Analyzed value (n = 10)
SRM NIST 8435 (Milk powder):
Moisture (g) 3.54 0.57 3.61 0.15
Protein (g) 25.86 0.67 25.30 0.14
Fat (g) 21.3 2.4 21.5 0.6
Ash (g) 5.97 0.11 5.98 0.04
SRM NIST 1849 (Infant/Adult Nutritional Formula):
Sugar (g) 49.8 1.8 49.7 0.49
Mg (mg) 157.8 6.9 159.7 1.7
Cu (mg) 2.03 0.04 1.98 0.03
Fe (mg) 17.71 0.33 17.70 0.29
Zn (mg) 15.23 0.51 15.68 0.36
4 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx

Table 1B 0.270.28 mg/100 g, respectively). Since this study is the first to

Percentage recovery of analysis of nutrient and toxic substances (n = 10 each). report these constituents, the values cannot be compared with
Analytical component Acceptance recovery Analyzed value those of other studies.
(%) (%)
Fructans (g/100 g) 98102* 100.4 0.8 3.2. Insecticide residues
Phytate (g/100 g) 98102* 101.9 4.5
Oxalate (mg/100 g) 90107* 95.6 5.1 Kaentawan has several advantageous characteristics over tradi-
Nitrate (mg/kg) 80110* 94.4 5.1
Cyanide (mg/kg) 80110* 102.9 3.9
tional plants including high growth rate, good tolerance to cold and
drought, the lowest fertilizer requirement for growth, and strong
Insecticide residues:
Alpha-endosulfan (mg/kg) 70120** 86.3 4.9
resistance to plant diseases and pests because generally Kaenta-
Beta-endosulfan (mg/kg) 70120** 87.5 6.8 wan has hairy oval shaped leaves and stems (Monti, Amaducci, &
Endosulfan-sulfate (mg/kg) 70120** 85.6 5.3 Venturi, 2005; Kays & Nottingham, 2008; Bach et al., 2015;
Lambda-cyhalothrin (mg/kg) 70120** 83.3 3.1 Slimestad, Seljaasen, Meijer, & Skar, 2010; Tassoni et al., 2010).
Permethrin (mg/kg) 70120** 84.2 4.0
Insecticide residues (organophosphorus, organochlorine, carba-
Cypermethrin (mg/kg) 70120** 95.8 9.6
Deltamethrin (mg/kg) 70120** 99.0 4.9 mate, pyrethroid) in fresh and boiled Kaentawan tubers were not
Diazinon (mg/kg) 70120** 84.6 3.5 detected (less than the detection limit) in almost all samples. Eigh-
Dicrotophos (mg/kg) 70120** 84.0 3.8 teen compounds of organophosphorus were studied including
Chlorpy-met (mg/kg) 70120** 84.6 7.7 dicrovos, methamidophos, mevinphos, diazinon, omethoate, dicro-
Piri-met (mg/kg) 70120** 82.1 7.9
tophos, monocrotophos, chlorpyrifos-methyl, dimethoate,
Chlorpy (mg/kg) 70120** 85.4 6.8
Malathion (mg/kg) 70120** 83.3 7.8 pirimiphos-methyl, chlopyrifos, parathion-methyl, pirimiphos-
Fenitrothion (mg/kg) 70120** 85.0 6.6 ethyl, malathion, and fenitrothion. All of these compounds had
Parathion (mg/kg) 70120** 84.4 8.7 limited detection (LOD) at 0.005 mg kg 1, whereas parathion-
Prothiofos (mg/kg) 70120** 88.5 6.3
ethyl, prothiofos, and profenofos had LOD at the level of
Ethion (mg/kg) 70120** 87.5 6.9
Triazophos (mg/kg) 70120** 82.3 8.4 0.01 mg kg 1. Four types of organophosphorus, namely, methi-
EPN (mg/kg) 70120** 88.3 7.4 dathion, ethion, triazophos, and ethyl p-nitrophenyl thionoben-
Oxamyl (mg/kg) 70120** 87.8 4.6 zenephosphonate (EPN), were included with LOD at
Methomyl (mg/kg) 70120** 92.8 4.7 0.01 mg kg 1. Four compounds of organochlorine, alpha-
Carbofuran (mg/kg) 70120** 88.0 3.4
endosulfan, beta-endosulfan, and endosulfan-sulfate had an LOD
Carbaryl (mg/kg) 70120** 89.6 4.8
Methiocarb (mg/kg) 70120** 88.6 4.2 at 0.0005 mg kg 1. Seven types of pyrethroid (bifentrin, lambda-
*
cyhalothrin, permethrin, cyfluthrin, cypermethrin, fenvalerate,
AOAC (2012).
** and deltamethrin) had LOD at 0.0005 mg kg 1. Only chlopyrifos
Codex Alimentarius Commission (2002) at concentration of P0.01 mg/kg to
60.1 mg/kg. was found in Kaentawan tuber with skin from Khonkaen province.
However, the level was lower than the Maximum Residue Level
(<0.01 mg kg 1 FW) as noted in the Thai Agriculture Standard
(0.05 mg kg 1 FW) (Thai Agriculture Standard, 2013). The source
Table 2
Nutrient compositions and toxic substances of fresh and boiled Kaentawan tuber.
of the insecticide possibly came from insecticide contaminated
soil, water, and/or air (Bruzzoniti, Sarzanini, Costantino, & Fungi,
Parameters (per 100 g edible fresh weight) Kaentawan tuber (Mean SD, 2006; Dasciana et al., 2011).
n = 4 each)
Peeled, Fresh Peeled, Boiled 3.3. Heavy metals
Edible (%) 82.5 1.2
Proximate composition: Heavy metal contents (Cd, Pb, As, and Hg) in all samples are
Energy (kcal) 10 1 92
shown in Table 3. Cadmium contents in all samples ranged from
Moisture (g) 78.0 0.9 80.2 1.7
Protein (g) 2.3 0.3 2.2 0.4 0.06 to 0.14 mg kg 1 FW. The average Cd level was slightly higher
Fat (g) 0.1 0.1 0.1 0.1 than the maximum contaminant contents stipulated by the Euro-
Ash (g) 1.0 0.2 1.0 0.2 pean Union (0.1 mg kg 1 for root vegetables and potatoes FW),
Carbohydrate (g) 18.6 1.0 17.5 1.9 though there is no specification of Cd for food in Thailand. Lead
Total Dietary fiber (g) 4.1 1.3 3.2 0.8
Fructans (g) 16.9 0.3 15.4 0.2
contents in all samples ranged from 0.20 to 0.47 mg kg 1 FW.
Sum of dietary fiber (g) 20.9 1.6 18.5 0.6 The average Pb level was lower than the level for food intended
Total sugars (g) 1.1 0.5 1.5 1.2 for direct human consumption as stipulated by the Thai Ministry
Minerals: of Public Health (1 mg kg 1 food). However, it was higher than
Calcium (mg) 20 4 20 4 the maximum levels of certain contaminants in foodstuffs as noted
Magnesium (mg) 16 1 16 2 by the European Union (0.1 mg kg 1 FW for root vegetables and
Phosphorus (mg) 74 7 72 10
potatoes). Arsenic content was very low in the studied samples
Sodium (mg) 30 1 28 6
Potassium (mg) 339 61 301 66 (ND). Mercury (Hg) was found only in peeled fresh Kaentawan
Iron (mg) 0.32 0.05 0.37 0.07 from Nakhon Ratchasima province (0.019 mg/kg FW), but it was
Copper (mg) 0.17 0.05 0.16 0.04 lower than the level of Hg for food intended for direct human con-
Zinc (mg) 0.27 0.04 0.28 0.04 sumption as stipulated by the Thai Ministry of Public Health
(0.5 mg kg 1 food).

3.1.2. Minerals 3.4. Nitrate and nitrite

In fresh and boiled Kaentawan tuber, potassium (301
339 mg/100 g FW) had the highest macro mineral content (Table 2).
Calcium, magnesium, phosphorus, and sodium (20, 16, 7274, and
2830 mg/100 g FW, respectively) were also presented. Trace ele-
ments were iron, copper, and zinc (0.320.37, 0.160.17, and
Nitrate content in all samples was lower than the quantitation
limit (<7.23 mg kg 1), whereas nitrite values were not detected
in all samples (<LOD, 0.39 mg kg 1) (Table 4). The average of
nitrate and nitrite values was lower than the level of nitrate and

Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx 5

Table 3
1
Heavy metal contents of Kaentawan tubers in Thailand, mg kg fresh weight.

Samples/Sources Cd Pb As Hg
LOD 0.001 0.017 0.145 0.010
LOQ 0.004 0.058 0.495 0.018
Kaentawan, peeled, fresh:
Nakhon Ratchasima 0.11 0.20 ND 0.019
Phetchabun 0.13 0.24 ND ND
Nakhon Pathom 0.13 0.29 ND ND
Khonkaen 0.13 0.32 ND ND
Mean SD 0.13 0.01 0.26 0.05 ND ND
Kaentawan, peeled, boiled:
Nakhon Ratchasima 0.11 0.29 ND ND
Phetchabun 0.06 0.25 ND ND
Nakhon Pathom 0.14 0.47 ND ND
Khonkaen 0.12 0.32 ND ND
Mean SD 0.11 0.03 0.33 0.10 ND ND

ND = not detectable (<LOD).

3.5. Oxalate, phytate, cyanide, and trypsin inhibitor activity

Table 4
1
Nitrate and nitrite contents of Kaentawan tubers in Thailand, mg/kg fresh weight. Oxalates in fresh and boiled samples ranged from 5 to
Sample/Sources Nitrate *
Nitrite** 22 mg/100 g FW (Table 5), which is less than 50 mg/100 g FW
Kaentawan, peeled, fresh:
when compared to several plants reported by Judprasong et al.
Nakhon Ratchasima 3.44 ND (2006). Foods with total oxalate levels over 50 mg/100 g FW are
Nakhon Pathom 2.46 ND classified as high oxalate foods (Boontaganon, Jhanno, & Savage,
Khonkaen 2.07 ND 2009; Chai & Liebman, 2005). Generally, oxalate is widely found
Phetchabun 6.33 ND
in small amounts in plants, such as blueberries, boysenberries,
Kaentawan, peeled, boiled: blackcurrants, red currants, red raspberries, black raspberries,
Nakhon Ratchasima 3.44 ND
and red tomatoes (26 mg/100 g FW). Rhubarb is high in oxalate,
Nakhon Pathom 2.46 ND
Khonkaen 2.07 ND up to 2751336 mg/100 g FW (Nguy n & Savage, 2013; Noonan
Phetchabun 6.33 ND & Savage, 1999). In Thailand, Judprasong et al. (2006) have
* 1 reported some plant foods that contain large amounts of oxalate
Nitrate values below limit of quantitation (7.23 kg FW).
**
ND = not detectable (<LOD, 0.39 mg kg 1 FW). (more than 100 mg/100 g FW), including acacia pennata, Chinese
convolvulus, bamboo shoot cultivated, coconut heart, peanut, and
soybean. They also reported a reduction in oxalate content due
nitrite in food (500 and 125 mg/kg, respectively) as recommended to cooking by boiling (loss more than 30%). This study found that
by the Thai Ministry of Public Health (2004). Plants accumulate oxalate content in all sources was reduced after boiling (45% from
nitrate in their roots, leaves, and stems and may lead to high levels. Nakhon Ratchasima and 75% from Khonkaen). The high percentage
Moreover, the concentration of nitrate and nitrite depends on of oxalate content reduction during boiling is probably due to its
types or varieties of plants, season, temperatures, light soil, types solubility in boiling water.
of fertilizer, and fertilizer use (EFSA, 2008). As mentioned earlier, Phytate content in all samples was low (0.100.19 g/100 g FW).
Kaentawan has several advantageous characteristics over tradi- In general, phytate is mainly found in cereal grains, legumes, and
tional plants, especially the very small fertilizer requirement for seeds (0.51.0 g/100 g FW), and in a lower concentration in tubers
growth [Monti et al., 2005; Kays & Nottingham, 2008; Slimestad and roots (0.050.10 g/100 g FW) (Lnnerdal, 2002; Reddy, 2002;
et al., 2010; Tassoni et al., 2010; Bach et al., 2015]. Phillippy, Lin, & Rascob, 2004). Phytate has a strong ability
to chelate metal ions including zinc, calcium, and iron

Table 5
Oxalate and phytate contents, cyanide, and trypsin inhibitor activity (per fresh weight, FW) of Kaentawan tubers in Thailand.

Samples/Sources Oxalate Phytate Cyanide Tyrpsin inhibitor activity

(mg/100 g FW) (g/100 g FW) (mg/k 1 FW) (TIU/mg)
LOD 0.9 0.03 2.79 0.15
LOQ 3.0 0.08 9.31 0.50
Kaentawan, peeled, fresh:
Nakhon Ratchasima 22.0 0.19 ND ND
Nakhon Pathom 7.0 0.14 ND ND
Khonkaen 21.0 0.18 ND ND
Phetchabun 7.2 0.15 ND ND
Kaentawan, peeled, boiled:
Nakhon Ratchasima 12.1 0.13 ND ND
Nakhon Pathom <LOQ 0.10 ND ND
Khonkaen 5.2 0.18 ND ND
Phetchabun <LOQ 0.14 ND ND

ND = not detectable (<LOD).

Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
6 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx
(Garca-Estepa, Guerra-Hernndez, & Garca-Villanova, 1999). Pre-
vious studies have reported food plants that contain high phytate
levels (0.082.1 g/100 g FW), including wheat grain, maize, rice,
cassava, potato, peanut, plantain, and lentils (Lazarte, Carlsson,
Almgren, Sandberg, & Granfeldt, 2015). Phillippy et al. (2004) have
reported phytate content in raw potato tubers in the range of 0.05
0.06 g/100 g FW. Consequently, Kaentawan tuber can be classified
as a food with low phytate content.
Cyanide content in the studied samples was very low (<LOD,
2.79 mg kg 1). Cyanogenic glycoside is widely distributed in the
plant and over 2500 plant species have been reported to contain
cyanogenic glycoside (Zagrobelny, Bak, & Moller, 2008). Generally,
cyanogens can be found throughout a plant, with large amounts in
leaves and roots, e.g., high in cassava and bamboo shoots (Cardoso
et al., 2005). Many cases of cyanide poisoning have been reported
for other plants, such as plum, apple, pear, bitter almonds, apricots,
and peaches (Ballhorn, Kautz, & Rakotoarivelo, 2009; Ballhorn,
2011, chap. 14).
Trypsin inhibitor activity was not detected in all Kaentawan
samples (LOD 0.15 trypsin inhibitor unit, TIU/mg). Most grain
legume seeds are major sources of trypsin inhibitors, such as soy-
bean (4384 TIU/mg), whereas barley, wheat, chick pea, lentil, pea,
and fava bean contain <25 TIU/mg (Guillamon et al., 2008).
4. Conclusion
Both fresh and boiled Kaentawan tubers grown in four major pro-
vinces in Thailand have beneficial nutrients and are good sources of
dietary fiber and fructans. They also contain low levels of several
chemical contaminants (insecticide residues; organophosphorus,
organochlorine, carbamate, and pyrethoid), heavy metals (cad-
mium, lead, arsenic, and mercury), and natural toxic substances
(nitrate, nitrite, cyanide, phytate, oxalate, and trypsin inhibitor). In
some Kaentawan tubers, toxic substances existed in terms of
chlopyrifos (insecticide residue) but in a very low amount
(<0.01 mg kg 1 FW), as well as lead (0.47 mg kg 1 FW) and cad-
mium (0.14 mg kg 1 FW). These levels are lower than those stipu-
lated for food intended for direct human consumption by the Thai
Ministry of Public Health. This studys results indicate that con-
sumption of Kaentawan from the four harvest provinces in Thailand
may not lead to any health problems. This information could be used
to promote Kaentawan as a healthy food for consumers.
Acknowledgements
We are grateful to the Thailand Research Fund, Thailand (RSA
5680039) for providing financial support, to the Institute of Nutri-
tion, Mahidol University for support all facilities and instruments
and to Mr. Attig A George for language manuscript editing.
References
AOAC (2012). Official method of analysis (19th ed.). Washington, DC: Association of
Officiating Analytical Chemists.
AOCS. (1997). American Oil Chemists Society, Ba 1275.
Bach, V., Clausen, M. R., & Edelenbos, M. (2015). Production of Jerusalem Artichoke
(Helianthus tuberosus L.) and Impact on Inulin and Phenolic Compounds (12
(pp. 97102). Kirstinebjergvej, Denmark: Department of Food Science, Aarhus
University.
Bai, Y., Zhou, L., & Wang, J. (2006). Organophosphorous pesticide residues in market
foods in Shaanxiarea, China. Food Chemistry, 98, 240242.
Ballhorn, D. J. (2011). Cyanogenic glycosides in nuts and seeds. Nuts & seeds in health
and disease prevention. (pp. 129136).
Ballhorn, D. J., Kautz, S., & Rakotoarivelo, F. P. (2009). Quantitative variability of
cyanogenesis in Cathariostachys madagascariensisthe main food plant of
bamboo lemurs in southeastern Madagascar. American Journal of Primatology,
71, 305315.
Boontaganon, P., Jhanno, E., & Savage, G. (2009). Total, soluble and insoluble
oxalate content of bran and bran products. International Journal of Food,
Agriculture and Environment, 7(34), 204206.
Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116
Bruzzoniti, M. C., Sarzanini, C., Costantino, G., & Fungi, M. (2006). Determination of
herbicides by solid phase extraction gas chromatography-mass spectrometry in
drinking water. Analytical Chimica Acta, 578(2), 241249.
Cardoso, A. P., Mirione, E., Ernesto, M., Massaza, F., Cliff, J., Haque, M. R., et al. (2005).
Processing of cassava roots to remove cyanogens. Journal of Food Composition
and Analysis, 18, 451460.
Chai, W., & Liebman, M. (2005). Oxalate content of legumes, nuts, and grain-based
flours. Journal of Food Composition and Analysis, 18(7), 723729.
Codex Alimentarius Commission. Report of the joint FAO/WHO Food standards
programme. Codex committee on residues of veterinary drugs in foods. CX/
RVDF 03/10, November 2002.
Dasciana, R., Tecia, C., Anayla, S., Vicente, S. N., Pierre, F., & Ronaldo, N. (2011).
Determination of Insecticide residues in Vegetal Fruits. Chromatgraphy Research
International, 6.
EFSA. (2008). Opinion of the Scientific Panel on Contaminants in food Chain on a
request from the European Commission to perform a scientific risk assessment
on nitrate in vegetables. The EFSA Journal 2008;69:179. Retrieved Sep 20 2015,
from: URL: http://www.efsa.europa.eu/EFSA/Scientific_Opinion/contamej_
689_nitrate_en.pdf.
Gao, Y., Shang, C., Saghai Maroof, M. A., Biyashev, R. M., Grabau, E. A., Kwanyuen, P.,
et al. (2007). A modified colorimetric method for phytic acid analysis in
soybean. Crop Science, 47, 17981803.
Garca-Estepa, R. M., Guerra-Hernndez, E., & Garca-Villanova, B. (1999). Phytic
acid content in milled cereal products and breads. Food Research International,
32, 217221.
Guillamon, E., Pedrosa, M. M., Burbano, C., Cuadrado, C., Sanchez, M., & Muzquiz, M.
(2008). The trypsin inhibitors present in seed of different grain legume species
and cultivar. Food Chemistry, 107, 6874.
Holloway, W. D., Argall, M. E., Jealous, W. T., Lee, J. A., & Bradbury, J. H. (1989).
Organic acids and calcium oxalic acid in tropical root crops. Journal of
Agricultural and Food Chemistry, 37, 337341.
Horton, H. R., Moram, L. A., Acrimgeour, K. G., Perry, M. D., & Rawn, J. D. (2006).
Principle of biochemistry (4th ed.). Upper Saddle River, NJ: Pearson Prentice Hall.
Jaworska, G. (2005). Content of nitrates, nitrites, and oxalates in New Zealand
spinach. Food Chemistry, 89, 235242.
Joye, D., & Hoebregs, H. (2000). Determination of oligofructose, a soluble dietary
fiber by high-temperature capillary gas chromatography. Journal of Association
Official of Analytical Chemistry International, 83, 10201025.
Judprasong, K., Charoenkiatkul, S., Sungpuag, P., Vasanachitt, K., & Nakjamanong, Y.
(2006). Total and soluble oxalate contents in Thai vegetables, cereal grains and
legume seeds and their changes after cooking. Journal of Food Composition and
Analysis, 19(4), 340347.
Judprasong, K., Tanjor, S., Puwastien, P., & Sungpuag, P. (2011). Investigation of Thai
plants for potential sources of inulin-type fructans. Journal of Food Composition
and Analysis, 2011(24), 642649.
Kays, S. J., & Nottingham, S. F. (2008). Biology and chemistry of Jerusalem Artichoke
(Helianthus tuberosus L.). Northwest, Florida: CRC Press.
Lazarte, C., Carlsson, N., Almgren, A., Sandberg, A., & Granfeldt, Y. (2015). Phytate,
zinc, iron and calcium content of common Bolivian food, and implications
for mineral bioavailability. Journal of Food Composition and Analysis, 39,
111119.
Lnnerdal, B. (2002). Phytic acid-trace element (Zn, Cu, Mn) interactions.
International Journal of Food Science and Technology, 37(7), 749758.
McGrath, S. P., Zhao, F. J., & Lombi, E. (2001). Plant and rhizosphere processes
involved in phytoremediation of metal-contaminated soils. Plant and Soil, 232,
207214.
Ministry of Public Health. (2004). Notification of the Ministry of Public Health, No.
281, B.E. 2547. Re: Food Additives.
Monti, A., Amaducci, M. T., & Venturi, G. (2005). Growth response, leaf gas exchange
and fructans accumulation of Jerusalem artichoke (Helianthus tuberosus L.) as
affected by different water regimes. European Journal of Agronomy, 23,
136145.
Nguy n, H. V. H., & Savage, G. P. (2013). Oxalate content of New Zealand grown and
imported fruits. Journal of Food Composition and Analysis, 31, 180184.
Nicholson, S. S. (2007). Nitrate and nitrite accumulating plants. Veterinary
Toxicology, 68, 876879.
Noonan, S. C., & Savage, G. P. (1999). Oxalic acid content of foods and its effect on
humans. Asia Pacific Journal of Clinical Nutrition, 8(1), 6474.
Onianwa, P. C., Adetola, I. G., Iwegbue, C. M. A., Ojo, M. F., & Tella, O. O. (1999). Trace
heavy metals composition of some Nigerian beverages and food drinks. Food
Chemistry, 66, 275279.
Phillippy, B. Q., Lin, M., & Rascob, B. (2004). Analysis of phytate in raw and cooked
potatoes. Journal of Food Composition and Analysis, 17, 217226.
Reddy, N. R. (2002). Occurrence distribution content and dietary intake of phytate.
Food Phytate, 2551.
Savage, G. P., Vanhanen, L., Mason, S. M., & Ross, A. B. (2000). Effect of cooking on the
soluble and insoluble oxalic acid content of some New Zealand foods. Journal of
Food Composition and Analysis, 13, 201206.
Slimestad, R., Seljaasen, R., Meijer, K., & Skar, S. L. (2010). Norwegian-grown
Jerusalem artichoke (Helianthus tuberosus L.): Morphology and content of sugars
and fructo-oligosaccharides in stems and tubers. Journal of the Science of Food
and Agriculture, 90, 956964.
Srinivasan, A., Giri, A. P., Harsullkar, A. M., Gatehouse, J. A., & Gupta, V. S. (2005). A
Kunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts anti-
metabolic effect on podborer (Helicoverpa armigera) larvae. Plant Molecular
Biology, 57, 359374.
 K. Judprasong et al. / Food Chemistry xxx (2016) xxxxxx 7

Tanjor, S., Judprasong, K., Chaito, C., & Jogloy, S. (2012). Inulin and
fructooligosacharides in different varieties of Jerusalem Artichoke (Helianthus
tuberosus L.). KKU Research Journal, 17(1), 2534.
Tassoni, A., Bagni, N., Ferri, M. M., Franceschetti, M., Khomutov, A., Marques, M. P.,
et al. (2010). Helianthus tuberosus and polyamine research: Past and recent
applications of a classical growth model. Plant Physiology and Biochemistry, 48,
496505.
Thai Agriculture Standard, TAS, 90022013 (2013). Pesticide residues: Maximum
residue limits. National Bureau of Agricultural Commodity and Food Standards.
Ministry of Agriculture and Cooperatives.
Ware, G. W., & Whitacre, D. M. (2004). The pesticide book (6th ed. p. 496).
Willoughby, OH: Meister Media Worldwide.
Zagrobelny, M., Bak, S., & Moller, B. L. (2008). Cyanogenesis in plants and
arthropods. Phytochemistry, 69, 14571468.

Please cite this article in press as: Judprasong, K., et al. Nutrients and natural toxic substances in commonly consumed Jerusalem artichoke (Helianthus
tuberosus L.) tuber. Food Chemistry (2016), http://dx.doi.org/10.1016/j.foodchem.2016.09.116