NFS Journal 23 (2021) 58–65

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NFS Journal
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Physicochemical properties and nutritional compositions of nipa palm

(Nypa fruticans Wurmb) syrup
Warasri Saengkrajang , Manat Chaijan , Worawan Panpipat *
Food Technology and Innovation Research Center of Excellence, Department of Food Science and Innovation, School of Agricultural Technology and Food Industry,
Walailak University, Thasala, Nakhon Si Thammarat 80161, Thailand

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

Keywords: Nipa palm (Nypa fruticans Wurmb.) syrups from three geographical plantation sites in Nakhon Si Thammarat,
Nipa palm Southern Thailand were characterized for their physicochemical properties and nutritional values. Variations in
Syrup color, clarity, viscosity, pH, total soluble solid, total acidity, salinity, water activity, browning intensity and 5-
Sweetener
hydroxymethylfurfural content of the syrups were observed among the plantation areas. The nutritional com­
Sugar
positions of the syrups also varied depending on their harvested locations. Sugar was the main dry matter
presented in all the nipa palm syrups, followed by protein and ash. The energy values of the syrups were in the
range of 376–413 kcal/100 g, dw. Sucrose was found as a major sugar in all syrup whereas glucose and fructose
were found as minor sugars. Low amounts of natural fructan, fructo-oligosaccharide, and kestose were found
only in one plantation area. The total phenolic acid and total flavonoid contents were determined in all syrup
with varying concentrations. Potassium and sodium were found as major elements. Phosphorus, magnesium,
silica, calcium, iron, copper, iodine, manganese, zinc, and chromium were also found at varying contents. Sugar
derivatives, non-protein nitrogenous compounds, polyphenols, organic acids, and some flavor substances were
identified in the nipa palm syrups by liquid chromatography-mass spectrometry. Results highlighted the varia­
tions in physicochemical properties and nutritional compositions of nipa palm syrup. Overall, the nipa palm
syrup can be regarded as an alternative sweetener for application in the functional food formulations.

1. Introduction
A high simple sugar consumption and a high-calorie sugar-to-diet
ratio are among the key risk factors for increasing a number of severe
diseases such as obesity, diabetes mellitus, metabolic syndrome, and
cardiovascular diseases [1–4]. To reduce the prevalence of obesity and
metabolic abnormalities attributed to added sugars, the development of
alternative sweeteners, both natural and artificial, have been intensively
studied. However, sweeteners have been continuously targets of scan­
dals and controversy, with allegations of carcinogenicity, fetal malfor­
mations, toxicity to liver and bladder, among other dangers. Although
many of these allegations have been researched and the sweeteners have
been considered as safe, a certain mistrust among consumers still re­
mains, adding to the fact that some are not allowed to be used in the USA
and other in the EU, the search for natural alternatives is important [5].
As a result, the natural unrefined syrups, which are recognized as safe,
have gained more attention. Among natural sweeteners, palm syrups
derived from Cocos nucifera (coconut palm), Borassus flabellifer (palmyra
palm), Arenga pinnata (sugar palm), Nypa fructicans (nipa palm), and
Phoenix dactylifera (date palm) have been used for a long period of time
[6–9]. Palm syrups generally contained higher nutrients and antioxi­
dants with a lower glycemic index than sugarcane [6–12].
Nipa palm (N. fructicans) is one of Arecaceae species, which naturally
grows in swamp and estuary areas throughout the Indian and Pacific
oceans [11,13,14]. It is also widely distributed in Thailand along the
mangrove forests and the largest plantation area (~ 41 km2) is in Pak
Panang River Basin, Nakhon Si Thammarat, Southern Thailand [9].
Typically, nipa palm can yield fresh sap from the inflorescences which
can be thermally concentrated to produce syrup [9]. Nipa palm syrup
represents the famous geographical natural product of Nakhon Si
Thammarat, Thailand. Traditionally, nipa palm syrup is prepared by
heating the fresh sap in an open pan over a wood-fired stove (>100 ◦ C)
with a continuous hand-blending until a viscous brown concentrated
sweet liquid with the total soluble solids of ≥65◦ Brix was obtained [9].
During the extended heating, numerous chemical reactions particularly
non-enzymatic browning reactions (e.g. Maillard and caramelization)

* Corresponding author.
E-mail address: pworawan@wu.ac.th (W. Panpipat).

https://doi.org/10.1016/j.nfs.2021.04.004
Received 30 January 2021; Received in revised form 27 April 2021; Accepted 27 April 2021
Available online 1 May 2021
2352-3646/© 2021 The Author(s). Published by Elsevier GmbH on behalf of Society of Nutrition and Food Science e.V. This is an open access article under the CC
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
W. Saengkrajang et al.
are generally taken place and influence the nutritional values and sen­
sory characteristics of the syrup.
It has been reported that the nutritional quality and sugar profile of
the palm syrup are principally governed by species, harvesting time,
plantation area, and processing [12,15,16]. Since nipa palm is typically
grown in the wetland with a moderate tidal influence, the nutrients in
the soil and water can definitely influence the growth performance and
the sap composition. At Pak Panang district, nipa palm plantations can
be found in fresh water, brackish, and saltwater regions. Also, the
heating time was adapted for a standard final product to reach the target
total solid content and was dependent on the composition of raw ma­
terial observed among regions. Therefore, this study aimed to investi­
gate the physiochemical properties and nutritional values of the nipa
palm syrups collected from three plantation sites in the Pak Panang
District, Nakhon Si Thammarat, Southern Thailand.
2. Materials and methods
2.1. Sap collection
The nipa palm saps used for the preparation of syrups were collected
from three different plantation sites in Pak Panang District, Nakhon Si
Thammarat, Southern Thailand during April 2020. Fifty palms per lot
were randomly chosen and three different lots (three days between lots)
of the sap were used. Freshly tapped nipa palm sap from the cut stalks of
inflorescences was collected in bamboo vessels in the presence of pieces
of Kiam (Cotylelobium lanceotatum craih) wood. The Kiam woods were
obtained from the same source and the weights of wood pieces were also
fixed in order to achieve a homogeneous treatment for samples from all
locations. The initial pH of the fresh sap harvested from the site I, II, and
III was 5.1 ± 0.0, 5.0 ± 0.0, and 5.6 ± 0.0, respectively, whereas the
total soluble solid (TSS) was 15 ± 0, 18 ± 0, and 19 ± 0◦ Brix, respec­
tively. The sap (80 L per lot) was packed in polyethylene bottles and
transported on ice in polystyrene foam boxes (4 ◦ C) to the laboratory
within 1 h.
All the sites are about 10 km away from the sea with 5–10 m above
the sea level. The site I is located at 8◦ 14′ 08.7′′ N 100◦ 13′ 31.0′′ E, Pak
Phraek sub-district. The site II is located at 8◦ 12′ 07.3′′ N 100◦ 14′ 10.5′′ E,
Khanap Nak sub-district. The site III is located at 8◦ 13′ 11.2′′ N
100◦ 14′ 47.5′′ E, Khanap Nak sub-district. For all sites, the plantations
were done naturally without the irrigation system and added fertilizer.
Regarding the plantation age, nipa palms from the site I and the site III
are around 15–16 years old whereas it is roughly 20–22 years old for the
site III. The pH of the soil/water at the site I, II, and III was 5.5 ± 0.0/4.6
± 0.0, 4.4 ± 0.0/5.1 ± 0.01, and 5.9 ± 0.0/5.4 ± 0.0, respectively. The
salinity of the soil/water at the site I, II, and III was 5 ± 0/101 ± 1, 7 ±
0/150 ± 0, and 5 ± 0/162 ± 1 ppt, respectively.
2.2. Preparation of nipa palm syrup
To prepare the syrup, the sap was heated using open-pan laboratory
scale evaporator at 110 ◦ C with continuous stirring (500 rpm) until the
TSS reached 70◦ Brix according to the Thai Industrial Standards Institute
(TISI) for syrup [17]. The time required to obtain the desired TSS was
220 ± 5, 190 ± 7, and 180 ± 3 min for the syrup from the site I, II, and
III, respectively. The syrup was cool down to room temperature
(27–30 ◦ C) and kept at 4–10 ◦ C until used. Three different lots of the
syrup were produced for each location to obtain triplications for all
analyses (n = 3).
2.3. Determination of physicochemical properties
The color values including lightness (L*), redness-greenness (a*) and
yellowness-blueness (b*) of the syrup were measured using Hunter Lab
(Color Flex, Hunter Associates Laboratory, VA, USA). The TSS was
measured using a hand refractometer (Atago, Japan). The intermediate
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NFS Journal 23 (2021) 58–65
browning product (IBP) and the browning intensity (BI) were observed
at 285 nm and 420 nm, respectively, using a Shimadzu UV-2100 spec­
trophotometer (Shimadzu Scientific Instruments Inc., Columbia, MD,
USA) [10]. The clarity was estimated by measuring the transmittance at
650 nm using a Shimadzu UV-2100 spectrophotometer. The viscosity
was measured using a Brookfield Viscometer Model RV-DV II Pro+
(Brookfield Engineering Inc., Middleborough, MA, USA) equipped with
the UL cylindrical spindle (ULA-15E). The water activity (aw) was
measured with an Aqualab Series 3TE aw meter (Devices Inc., Pullman,
WA, USA). The pH was determined using a pH meter (Cyberscan 500,
Singapore). The total acidity was determined by titration method and
report as percentage of acetic acid [18]. The salinity was measured by a
hand salinometer (Atago, Japan). The content of 5-hydroxymethylfurfu­
ral (HMF) was analyzed using the high performance liquid chromatog­
raphy (HPLC) [19]. All the analyses were done at room temperature.
2.4. Determination of proximate composition and energy value
The contents of moisture (AOAC method number 950.46), crude
protein (AOAC method number 928.08, Kjeldahl factor of 6.25), fat
(AOAC method number 963.15), and ash (AOAC method number
920.153) of the nipa palm syrups were analyzed according to AOAC
[18]. The carbohydrate content was obtained by the difference. The
energy value of the nipa palm syrup was measured in a bomb calorim­
eter [18].
2.5. Determination of total sugar, reducing sugar, and sugar profile
The total sugar content was estimated by the phenol‑sulfuric acid
method [20]. The reducing sugars were quantified by colorimetric
method with 3,5-dinitrosalicylic acid reagents, as described by Miller
[21]. The sugar profile was analyzed using the HPLC (Shimadzu, CR6A
Chromatopac, Kyoto, Japan) with a Zorbax carbohydrate column (4.6 ×
150 mm, 5 μm size) and refractive index detector [9]. The solution of
acetonitrile and water (76:24) was used as the mobile phase at a flow
rate of 0.8–1 mL/min and injection volume 50 μL. The samples were
prepared by making appropriate dilutions with distilled water. All
sample solutions were passed through a 0.45-μm syringe filter (nylon) to
remove particulates prior to HPLC analysis. Standard curves for D-
glucose, D-fructose, and sucrose were prepared at concentrations
ranging from 1 to 10% (w/v). The chromatographic peaks corresponding
to each sugar were matched with the retention time of the standard. A
calibration curve fitted by linear regression analysis was prepared using
standards to determine the relationship between the peak area and
concentration. The unidentified peaks were assigned as unidentified
sugars. In addition, fructan, and fructo-oligosaccharides including kes­
tose (GF2), nystose (GF3), and fructosyl nystose (GF4) were determined
using the HPLC [22].
2.6. Determination of mineral profile
The minerals including potassium (K), sodium (Na), calcium (Ca),
magnesium (Mg), copper (Cu), iron (Fe), manganese (Mn), zinc (Zn),
phosphorus (P), iodine (I), chromium (Cr) and silica (Si) were deter­
mined using inductive couple plasma-optical emission spectrometry
(ICP-OES, Perkin-Elmer model optima 3300 Dv, Norwalk, CT, USA)
according to the AOAC method [18].
2.7. Determination of total phenolic content (TPC) and total flavonoid
content (TFC)
The TPC was determined by the Folin-Ciocalteu method as described
by Kahkonen et al. [23]. The syrup (0.3 mL) was mixed with Folin-
Ciocalteu reagent (Sigma-Aldrich, St. Louis, MO, USA) (1:9 with
water), followed by the addition of 1.2 mL of 7.5% sodium carbonate
solution (Sigma-Aldrich). The mixture was kept in the dark for 30 min
W. Saengkrajang et al. NFS Journal 23 (2021) 58–65

Table 1
Appearances and some physicochemical characteristics of nipa palm syrups obtained from different plantation areas.
Parameters Site I Site II Site III

Appearance

Color
L* 8.64 ± 0.08a 13.21 ± 0.27b 15.75 ± 0.22c
a* 7.07 ± 0.01c 4.87 ± 0.16b 1.74 ± 0.30a
b* 5.78 ± 0.08a 9.73 ± 0.16b 12.50 ± 0.40c
Total soluble solid (◦ Brix) 70±1ns 70±0ns 70±1ns
Browning index
IBP (A285) 165 ± 2c 122 ± 5b 72 ± 1a
BI (A420) 12 ± 0c 5 ± 0a 7 ± 0b
Clarity (%) 86±4ns 89±1ns 85±1ns
Viscosity (cp) 547 ± 3c 255 ± 1a 351 ± 2b
Water activity 0.80 ± 0.00ns 0.80 ± 0.00ns 0.80 ± 0.01ns
pH 5.1 ± 0.0a 5.1 ± 0.0a 5.4 ± 0.0b
Total acidity (%) 0.5 ± 0.0c 0.4 ± 0.0b 0.3 ± 0.0a
Salinity (%) 68 ± 1a 72 ± 1b 78 ± 2c
HMF (mg/kg) 18 ± 0b 5 ± 0a 5 ± 0a

Values are given as mean ± standard deviation from triplicate determinations (n = 3).
Different letters in the same row indicate significant differences (p < 0.05).
IBP = Intermediate browning product.
BI = Browning intensity.
HMF = 5-Hydroxymethylfurfural.
ns
Not significant.

and then the absorbance was measured at 725 nm using a Shimadzu UV-
2100 spectrophotometer. The TPC was expressed as mg gallic acid
equivalent (GE)/g dry weight (dw) of the sample.
The method of Woisky and Salatino [24] was used to determine the
TFC. In brief, 0.5 mL of syrup was mixed with 95% ethanol (Sigma-
Aldrich). Thereafter, 0.1 mL of 10% aluminum chloride (Sigma-
Aldrich), 0.1 mL of 1 M potassium acetate (Sigma-Aldrich), and 2.8 mL
of distilled water were added, thoroughly mixed, and kept at room
temperature for 30 min. Subsequently, the absorbance was read at 415
nm using a Shimadzu UV-2100 spectrophotometer. The TFC was
expressed as mg rutin equivalent (RE)/100 g, dw.
2.9. Statistical analysis
A completely randomized design was used in this study. All results
were expressed as mean ± standard deviation. The data were statisti­
cally performed by the one-way analysis of variance. Comparison of
means was carried out by Duncan’s multiple-range test to identify sig­
nificant differences (p < 0.05) among treatments [25], using the Sta­
tistical Package for Social Science (SPSS 23.0 for windows, SPSS Inc.,
Chicago, IL, USA).
3. Results and discussion

2.8. Liquid chromatography-mass spectrometry (LC-MS) identification 3.1. Physicochemical properties

Chemical constituents in the nipa palm syrups obtained from
different plantation areas were identified by using HPLC (Agilent 6200
series, USA) coupled with TOF/Q-TOF mass spectrometer (model
G6545A). The sample (10 mg) was diluted with 1 mL distilled water and
centrifuged at 10,000 ×g at 4 ◦ C for 10 min. Sample (10 μL) was injected
with auto samplers needle wash system (G7129B). The system was
programed as followed: binary pump (G7120A) at 50 min with post time
of 5 min, flow rate of 200 μL/min, maximum flow gradient of 100 mL/
min and maximum pressure of 800 bar. The mobile phase was a mixture
of water (solvent A) and acetic acid, 2% (v/v) aqueous solution (solvent
B) at a ratio of 95:5. The chromatographic separation column temper­
ature was set at 25 ◦ C. The eluent was monitored with the diode array
detector (DAD-model G7117A) at the wavelength of 257, 278, 320, and
340 nm. The dual Agilent jet stream electrospray ionization (Dual AJS
ESI negative mode) was coupled with nitrogen gas (drying gas) to obtain
simultaneously mass spectra. Online database and TOF LC-MS data
processer (6200 series TOF/6500 series Q-TOF B.08.00) were used for
qualitative identification of chemical constituents in the syrup.
Physicochemical characteristics of the nipa palm syrups obtained
from three different plantation areas including color, TSS, browning
index, clarity, viscosity, aw, pH, total acidity, salinity, and HMF content
are shown in Table 1. All of the samples appeared in a transparent dark
brownish color (Table 1) with the L* values ranging from 8.64–15.75,
the a* values ranging from 1.74–7.07, and the b* values ranging from
5.78–12.50. The difference may be caused by the difference in heating
time to gain the target TSS of 70◦ Brix required by TISI. The longest
heating time was noticeable in the syrup obtained from the site I (220
min), followed by the site II (190 min) and the site III (180 min). The
heating time was mainly influenced by the original TSS content of the
corresponding sap. The sap harvested from the site I had the lowest TSS
content of 15◦ Brix, followed by the site II (18◦ Brix), and the site III
(19◦ Brix). Generally, sugar was the main dry matter of the nipa palm
sap, followed by ash and protein [9]. During thermal concentration,
non-enzymatic browning reactions particularly Maillard reaction and
caramelization are generally taken place [26,27], resulting in the com­
plex changes in color, flavor, and appearance of the final syrup. The
results were in agreement with the IBP and BI of syrups (Table 1), in
which the highest values were found in the syrup from the site I,

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Table 2
Proximate composition, sugar profile, energy value, total phenolic content, and total flavonoid content of nipa palm syrups obtained from different plantation areas.
Composition Site I Site II Site III

Proximate composition
Moisture (g/100 g, fw) 28.9 ± 0.0a 31.5 ± 0.1c 30.0 ± 0.0b
Protein (g/100 g, dw) 2.0 ± 0.0a 2.9 ± 0.1c 2.4 ± 0.1b
Ash (g/100 g, dw) 3.8 ± 0.0a 4.2 ± 0.0b 3.8 ± 0.0a
Fat (g/100 g, dw) ND ND ND
Fiber (g/100 g, dw) ND ND ND
Carbohydrate (g/100 g, dw) 94.2 ± 0.0c 92.9 ± 0.1a 93.7 ± 0.0b
Total sugar (g/100 g, dw) 93.5 ± 1.1ns 93.6 ± 3.7ns 93.1 ± 1.5ns
Reducing sugar (g/100 g, dw) 19.6 ± 0.0c 10.0 ± 0.0a 14.2 ± 0.0b
Sugar profile (g/100 g, dw)
Glucose 9.8 ± 0.1c 4.5 ± 0.0a 7.6 ± 0.1b
Fructose 9.7 ± 0.3c 4.6 ± 0.1a 5.8 ± 0.0b
Sucrose 64.6 ± 0.1a 80.5 ± 0.4c 73.9 ± 0.2b
Fructan 0.04 ± 0.01 ND ND
Fructo-oligosaccharides 0.04 ± 0.01 ND ND
Kestose 0.04 ± 0.01 ND ND
Nystose ND ND ND
Fructosyl nystose ND ND ND
Unidentified sugars 9.2 ± 1.0b 5.9 ± 1.2a 5.7 ± 1.2a
Energy (kcal/100 g, dw) 376 ± 1a 413 ± 1c 394 ± 1b
Total phenolic content (mg GE/100 g, dw) 30.3 ± 0.2b 44.3 ± 0.1c 16.9 ± 0.2a
Total flavonoid content (mg RE/100 g, dw) 27.2 ± 0.3c 18.2 ± 0.5b 16.3 ± 0.1a

Values are given as mean ± standard deviation from triplicate determinations (n = 3).
Different letters in the same row indicate significant differences (p < 0.05).
ND = not detected.
GE = gallic acid equivalent.
RE = rutin equivalent.
ns
Not significant.

followed by the site II, and the site III (p < 0.05). Melanoidins formed at
the final stage of non-enzymatic browning reactions [28,29] were
attributed mainly to the color formation of the syrup. In addition, the
polyphenol oxidase activity in the fresh sap may cause the browning of
the unheated sap to some degree. The results suggested that the
browning intensity of the final syrup was dominated by the plantation
area and the heating time. No significant difference in clarity was
observed in all syrups (p > 0.05) which were in the range of 85–89%
(Table 1). According to Naknean and Meenune [30], all the nipa palm
syrups can be classified as high grade syrup due to their high clarity
values. The highest viscosity was observed in the syrup obtained from
the site I (547 cp), followed by the site III (351 cp), and the site II (255
cp) (Table 1). This was in agreement with the carbohydrate, glucose, and
fructose contents of those syrups (Table 2). Rheological behavior of
syrups was governed by the type of sugar presented [31,32].
The aw of nipa palm syrups obtained from 3 different plantation
areas were not different (aw = 0.80) (p > 0.05) (Table 1). This may be
due to the same TSS presented in all syrups (Table 1). The aw of the
syrups in this study were comparable to the report of Phetrit et al. [9],
who found the aw of 0.79 in the nipa palm syrup. Kongkaew et al. [10]
suggested that the aw was one of the factors determining the storage
stability of the syrup.
Mild acidic pH was noted in all syrups ranging from 5.1–5.4
(Table 1). The highest pH value was observed in the syrup collected from
the site III (p < 0.05). No significant difference in pH was detected in the
syrups obtained from the site I and the site II (p > 0.05). This result was
somehow in agreement with the total acidity detected in the syrups,
which were in the range of 0.3–0.5% (Table 1). The pH of nipa palm
syrup in this study was higher than raffia palm syrup (pH 3.8) and oil
palm syrup (pH 3.5) reported by Oboh et al. [33]. The variation in the
pH might be due to the differences in the type and the content of acidic
substances. Numerous organic acids such as succinic acid, tartaric acid,
malic acid, citric acid, and lactic acid were previously detected in the
nipa palm sap [34]. Naknean and Meenune [30] suggested that the
organic acids were concentrated during palm syrup preparation,
resulting in the decrease in the pH of the final syrup. Certain organic
acids can also be generated via Maillard reaction [35,36], which can
influence the acidity of the final syrup. However, the natural buffering
system in the original sap may control the rate of pH reduction upon
syrup preparation. The salinity of nipa palm syrups ranged from 68 to
78% (Table 1). The highest salinity was found in syrup obtained from
the site III (p < 0.05), followed by those from the site II, and the site I.
The saltiness of foods is generally due to the presence of sodium chlo­
ride, potassium chloride, and calcium chloride [37].
The HMF contents of nipa palm syrups obtained from 3 different
plantations are shown in Table 1. The highest HMF content (18 mg/kg)
was observed in the syrup collected from the site I (p < 0.05). While, the
syrups harvested from the site II and the III had a low HMF content (~5
mg/kg). It was likely that heating for longer periods of time resulted in
the greater HMF content. Shapla et al. [38] reported a relationship be­
tween HMF concentration and heating time. Although humans may gain
between 30 and 150 mg HMF daily from foods, the safe levels of HMF
intake are not well defined [38]. HMF is a furan derivatives containing
both aldehyde and alcohol functional groups usually formed by thermal
induced reducing sugar degradations through the Maillard reaction and
caramelization [12,38]. HMF is considered as carcinogen due to its
cytotoxicity, mutagenicity and potential to cause chromosomal aberra­
tion [39–43], which may link to the safety issue of the syrup. HMF has
been shown to induce and promote aberrant crypt foci (ACF, preneo­
plastic lesions) in rat colon [40,41]. A significant dose-related increase
of ACF in rats after oral administration of a single dose of 0–300 mg/kg
body wt of HMF was reported by Zhang et al. [42]. Surh et al. [43]
indicated the induction of skin papillomas after topical administration of
10–25 μmol HMF to mice. However, numerous positive effects of HMF
have been reported [37]. For instance, Zhao et al. [44] reported the in
vitro antioxidant and antiproliferative activities of HMF. Yamada et al.
[45] reported the anti-allergic activity of HMF in basophilic cells. Li
et al. [46] showed the protective role of HMF against hypoxic injury. In
addition, the HMF metabolites are rapidly eliminated in urine [47].

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3.2. Proximate composition Table 3

Elemental composition of nipa palm syrups obtained from different plantation
The proximate compositions of the nipa palm syrups obtained from 3 areas.
different plantation areas are shown in Table 2. The moisture contents of Elements (mg/kg, dw) Site I Site II Site III
all syrups were in the range of 28.9–31.5% (Table 2), which were lower Potassium (K) 10,260 ± 240a 12,500 ± 280b 10,480 ± 90a
than the previous report of Phetrit et al. [9] on nipa palm syrup (40.3%, Sodium (Na) 4740 ± 20c 3400 ± 120a 4500 ± 60b
fw). The difference was probably due to the difference in the final TSS of Phosphorus (P) 520 ± 20a 710 ± 10c 590 ± 30b
the syrup. Here, the final TSS contents were set at 70◦ Brix whereas the Magnesium (Mg) 290 ± 10a 340 ± 10b 260 ± 20a
Silica (Si) 210 ± 4a 430 ± 3c 270 ± 10b
value of 65◦ Brix was reported by Phetrit et al. [9]. Carbohydrates were
Calcium (Ca) 30 ± 1a 60 ± 4b 30 ± 1a
the most abundant component in all nipa palm syrups, which were in the Iron (Fe) 15 ± 1a 23 ± 1b 16 ± 1a
range of 92.9–94.2% (dw). The syrup collected from the site I contained Copper (Cu) 2 ± 0.1ns 2 ± 0.1ns 2 ± 0.1ns
a higher carbohydrate content (94.2%) than the site III (93.7%) and the Iodine (I) 3 ± 0c 1 ± 0a 2 ± 0b
site II (92.9%) (p < 0.05). Protein contents in all samples were in the Manganese (Mn) 2 ± 0.1b 2 ± 0.1b 1 ± 0.0a
Zinc (Zn) 1 ± 0.1a 2 ± 0.1b 1 ± 0.1a
range of 2.0–2.9% (dw). The protein content of the syrup obtained from Chromium (Cr) 0.1±0ns 0.1±0ns 0.1±0ns
the site II (2.9%) was slightly higher than those of the site III (2.4%) and
the site I (2.0%) (p < 0.05). The ash contents of the syrup were in the Values are given as mean ± standard deviation from triplicate determinations (n
= 3).
range of 3.8–4.2% (dw), in which the sample from the site II had the
Different letters in the same row indicate significant differences (p < 0.05).
highest ash content (p < 0.05). The values were about 2-fold greater
dw = dry weight.
than the ash content in palm syrup reported by Luis et al. [48]. Lipid and ns
Not significant.
fiber were not detected in all nipa syrups (Table 1). In comparison to
Phetrit et al. [9], there were some variations in the proximate compo­
Meenune [12]. The glucose and fructose naturally occurred in the
sition of the nipa palm syrup. The carbohydrate, ash, and protein con­
original sap and might be added up in the syrup via the sugar inversion
tents were previously found to be 96.5, 3.0, and 0.4%, respectively [9].
during the heating process. From the results, the releasing of glucose and
This was probably due to the variations in the intrinsic factors and the
fructose during heating should be taken place since different contents of
processing conditions, such as the plantation area, the season of harvest,
sucrose, glucose and fructose were found in three locations with
the method of syrup preparation, and the target concentration.
different heating times (Table 2). For example, the highest glucose and
fructose contents with the lowest sucrose content was found in syrup
3.3. Sugar content, sugar profile, and energy value
from the site I in which the longest heating time was applied. It should
be noted that the fructan (0.04%, dw), fructo-oligosaccharides (FOS;
The total sugar content, reducing sugar content, and sugar profile of
0.04%, dw) and ketose (0.04%, dw) were detected in syrup collected
nipa palm syrups obtained from 3 different plantation areas are pre­
from the site I, whereas no fructan, FOS and ketose were identified in the
sented in Table 2. Sugars are the most predominant dry matter presented
syrups from the other sites (Table 2). Nystose and fructosyl nystose were
in all nipa palm syrups. No differences in the total sugar contents among
also not detected in all samples. FOS are naturally occurring sugars with
syrups were noticeable (93.1–93.6%) (p > 0.05). These values were
potentially beneficial nutritional effects which are widely distributed
close to nipa palm syrup (96%) [9] but there were higher than those in
throughout the plant kingdom [51]. FOS constitute a series of homolo­
palm tree syrup (66%) [48] and palmyra palm syrup (80%) [12]. It can
gous oligosaccharides derived from sucrose usually represented by the
be implied that the total sugar content in the syrup was dominated by
formula GFn, which are mainly composed of 1-kestose (GF2), nystose
the botanical species and environmental growth condition. Theer­
(GF3), and fructofuranosyl nystose (GF4), in which two, three, and four
awitaya et al. [49] demonstrated that nipa palm grown in the brackish
fructosyl units are bound at the β-2,1 position of glucose, respectively
water or the salt water area tended to produce more sugar, due to the
[52]. FOS have a low sweetness intensity (approximately one third as
adaptation of this plant in the salt stress condition. However, in this
sweet as sucrose) and supply small amounts of energy (0–3 kcal/g) [52].
study, the soil/water salinity is not correlated with the total sugar
FOS can also be served as prebiotics because FOS cannot be digested by
content. Interestingly, the reducing sugar contents in the syrups varied
the enzymes of the small intestine and they are fermented in the large
depending on the plantation site, where the syrup from the site I had the
intestine to selectively stimulate the growth of probiotic-like bacteria
highest reducing sugar content (19.6%), followed by the site III (14.2%),
[53]. However, some unidentified sugars (5.7–9.2%, dw) were observed
and the site II (10.0%) (p < 0.05). Typically, the reducing sugars are
in all samples (Table 2). The energy values obtained from all the syrups
more reactive to various chemical reactions such as Maillard reaction
are listed in Table 2. The highest energy was found in syrup collected
than the non-reducing sugars [50]. Also, monosaccharides like glucose
from the site II (413 kcal/100 g), followed by those from site III (394
are more prone to Maillard reaction than the disaccharides. It can be
kcal/100 g), and the site I (376 kcal/100 g) (p < 0.05). Since all the
implied that the syrup from the site I may undergo higher browning
samples had similar concentration of the total sugar, the variation of
reaction than other syrups during storage.
energy was possibly linked to the type of sugar and its individual con­
For the sugar profile, the nipa palm syrups were composed mainly of
centration as well as the presence of other sources of energy like proteins
sucrose (64.6–80.5%, dw), followed by glucose (4.5–9.8%, dw), and
and alcohols.
fructose (4.6–9.7%, dw) (Table 2). The results were in agreement with
Phetrit et al. [9] who found the contents of sucrose, glucose, and fructose
of 78.3% (dw), 9.8% (dw), and fructose 4.8% (dw), respectively, in the 3.4. TPC and TFC
nipa palm syrup. Among the plantation area, nipa palm syrup collected
from the site II contained the highest amount of sucrose (80.5%, dw), The TPC of nipa palm syrups obtained from the 3 different
followed by the site III (73.9%, dw) and the site I (64.6%, dw) (p < 0.05). geographical plantations were in the range of 16.9–44.3 mg GE/100 g,
The similar trends were noticed for the glucose and fructose concen­ dw (Table 2). The highest TPC was found in the syrup harvested from the
trations, in which the highest amounts of both sugars were detected in site II (44.3 mg GE/100 g), followed by those from the site I (30.3 mg
the syrup from the site I (glucose = 9.8%, dw and fructose = 9.7%, dw), GE/100 g) and the site III (16.9 mg GE/100 g) (p < 0.05). For the TFC
followed by those from the site III (glucose = 7.6%, dw and fructose = (Table 2), the highest value was found in the syrup obtained from the
5.8%, dw), and the site II (glucose = 7.6%, dw and fructose = 5.8%, dw) site I (27.2 mg RE/100 g), followed by those from the site II (18.2 mg
(p < 0.05). The contents of glucose and fructose in the nipa palm syrups RE/100 g) and the site III (16.3 mg RE/100 g) (p < 0.05). The results
were comparable to the palmyra syrup reported by Naknean and were in agreement with the study of Willis et al. [54] who found the

62
W. Saengkrajang et al. NFS Journal 23 (2021) 58–65

Table 4
Proposed structures of chemical constituents based on liquid chromatography-mass spectrometry analyses in nipa palm syrups obtained from different plantation
areas.
Compounds Diff range (DB, ppm) Molecular formula m/z (observed) Retention time (min) Score (DB, %)

Site I Site II Site III

Alpha,beta-Trehalose − 4.39-(− 1.33) C12H22O11 365.11 1.92–2.19 90.92 98.87 92.30

Bis-D-fructose 2′ ,1:2,1′ -dianhydride − 0.17-0.45 C12H20O10 347.10 1.96–1.97 99.35 99.21 99.3
Maltotriose − 0.94-0.67 C18H32O16 527.16 1.99–2.12 98.05 98.07 98.44
α-D-Glucose − 2.23-(− 0.09) C6H12O6 203.05 1.82–2.06 98.52 96.76 98.84
Sucrose − 0.2-0.72 C12H22O11 365.11 2.21–2.93 99.89 98.84 99.11
Glutathione − 0.24-0.63 C10H17N3O6S 308.09 2.45 ND 98.42 98.59
Epidermin 0.49 C11H19NO6 262.13 2.40 99.62 ND ND
N-(1-Deoxy-1-fructosyl)valine − 0.98 C11H21NO7 280.14 2.44 99.38 ND ND
Acetyl-maltose − 0.23-0.89 C14H24O12 407.12 2.56–2.60 93.47 99.10 99.45
Medicagol 0.51–0.77 C16H8O6 297.04 2.62 ND 90.32 90.12
N-Tris[hydroxymethyl]methyl-2 aminoethanesulfonic − 2.20-(− 2.49) C6H15NO6S 230.07 2.99–3.11 ND 90.92 91.29
acid
Meconic acid 2.79-(− 3.20) C7H4O7 222.98 3.20–3.29 90.50 90.58 92.27
2,3-Butanediol glucoside 0.99 C10H20O7 275.11 3.61 ND 99.63 ND
3-Amino-2-naphthoic acid 0.97–1.78 C11H9NO2 188.07 11.83–11.96 ND 99.50 97.67
Ser Ser Arg − 0.65-(− 1.66) C12H24N6O6 349.18 16.29–16.36 97.49 97.29 97.96
Asp Asn Lys − 1.30-(− 2.05) C14H25N5O7 393.21 17.76–17.82 97.00 96.85 97.46
Arg Gln Arg − 3.83-(− 3.05) C17H34N10O5 481.26 20.10 92.29 90.98 92.93
Cis-Miyabenol C 0.08 C42H32O9 681.21 24.43 ND 97.63 ND
Pentyl heptanoate 1.20 C12H24O2 218.21 25.85 ND ND 99.56
3-(L-Menthoxy)-2-methylpropane-1,2-diol − 0.94-0.19 C14H28O3 262.24 26.50–26.52 99.10 ND 99.6
Decyl isobutyrate − 0.18-0.83 C14H28O2 246.24 30.90–30.96 99.86 99.93 99.71
(− )-Euphomine 1.28–2.35 C32H44O8 579.29 46.19–46.20 98.14 98.18 96.00
6-hydroxy buspirone 0.77–2.12 C21H31N5O3 419.28 46.84 98.96 98.59 97.76
Montanol 2.13–2.53 C21H36O4 375.25 46.86–46.89 96.90 ND 96.16

ND = not detected.

significant differences in the TPC and TFC among the Madhura’s sweet
sorghum (Sorghum bicolor L. Moench) syrups harvested from different
sites in Kenya. It has been reported that the TPC and TFC of the nipa
palm syrup collected from local producers in Pak Phanang River Basin,
Southern Thailand in April 2018, was found to be 190.1 mg GE/100 g
(dw) and 42.7 mg RE/100 g (dw), respectively [9]. The difference may
be due to the geographical and seasonal variations in the secondary
metabolic products which may relate to the environment condition each
year. Sampaio et al. [55] reported that the environmental conditions
significantly influenced the production and accumulation of the primary
and secondary metabolites. The distribution of the metabolites e.g.
sugars and phenolics in the inflorescence and root parts of Tithonia
diversifolia were governed by the variation of some soil nutrients [55].
The highest acidity and saltiness of water and soil of the site II may relate
to the highest TPC of the syrup collected from that site. Both endogenous
and exogenous polyphenols may influence the TPC and TFC of the nipa
palm syrups. The endogenous polyphenols and flavonoids accumulated
in the inflorescences of the nipa palm can be released into the sap during
harvesting. The exogenous polyphenols can be derived from pieces of
Kiam wood which were intently added as a natural preservative for fresh
sap [9,56].
3.5. Elemental composition
Table 3 presents the elemental compositions in the nipa palm syrups
collected from 3 different geographical plantations. The major minerals
in nipa palm syrups were K (10260–12,500 mg/kg) and Na (3400–4740
mg/kg). Since soil particles are negatively charged in nature, the min­
eral cations such as K and Na are largely adsorbed to the negatively
charged surface of the soil particles, leading to their high availability to
plant roots [57]. Reid and Hayes [58] suggests that K is the most
abundant cation in the plant cells and essential for plant growth. As a
result, it can be found in the nipa palm syrup in the largest quantity.
Moreover, the availability of the nutrients on the root surface are
determined by the chemical (e.g., surface charge and pH) and physical
(e.g., structure and texture) properties of the soil, the environmental
growth conditions, the season, and the plant metabolism [57]. The
highest K content was found in the syrup from the site II (p < 0.05),
followed by those from the site I and III. Interestingly, the largest sodium
accumulation was noticed in syrup obtained from the site I, followed by
those from the site III, and the site II (p < 0.05). The soil and water
salinity of the site I were low-moderate but the syrup from this site had
the highest Na content. The minor elements in all syrups were P
(520–710 mg/kg), Mg (260–340 mg/kg), and Si (210–430 mg/kg),
whereas Ca (20–60 mg/kg), Fe (15–23 mg/kg), Cu (2 mg/kg), I (1–3
mg/kg), Mn (1–2 mg/kg), Zn (1–2 mg/kg), and Cr (0.1 mg/kg), which
were found as trace elements (Table 3). From the results, the minerals
were varied considerably among the samples. The syrup collected from
the site II tended to show the highest content for all minerals, except Na
and I. This result was in line with the ash content of the syrup from the
site II (Table 2). Naturally, nipa palm is a mangrove palm, which grows
well in the mangrove environment and moderately saline sites like
shallow lagoons or estuaries [49,59]. As a consequence, a higher total
mineral content could be expected in the nipa palm sap when compared
to other land palms such as coconut and palmyra palm. Phetrit et al. [9]
reported that K was the most abundant element in the nipa palm syrup
and other elements such as Na, P, Mg, Ca, Fe, Cu, Zn, Mn, and I were also
detected. Interestingly, Si was found in nipa palm syrups in which the
syrup obtained from the site II presented the highest concentration. Si is
descried as a valuable trace element having various health benefits such
as bone strengthening, improvement the collagen production, reduction
the risk of atherosclerosis, and anti-inflammatory activity [60].
3.6. Profiling of chemical constituents in nipa palm syrup
Several chemical constituents in nipa palm syrups analyzed by LC-
MS are given in Table 4. The chemical compounds in nipa palm
syrups were identified on the basis of m/z values. From the results,
several chemical compounds were found in nipa palm syrups such as
sugars and their derivatives, non-protein nitrogenous compounds, and
polyphenols. Alpha, beta-trehalose, bis-D-fructose 2′ ,1:2,1′ -dianhydride,
maltotriose, α-D-glucose, sucrose, acetyl-maltose, and 2,3-butanediol
glucoside were mainly observed as sugar derivatives in nipa palm
syrup (Table 4). Sucrose and glucose were principally identified and

63
W. Saengkrajang et al.
highly correlated with the sugar profile in Table 2. The minor sugar
derivatives e.g. alpha, beta-trehalose, bis-D-fructose 2′ ,1:2,1′ -dianhy­
dride, maltotriose, acetyl-maltose, and 2,3-butanediol glucoside may
contribute to the unidentified sugars presented in Table 2. For the non-
protein nitrogenous compounds, N-tris[hydroxymethyl]methy, l-2
aminoethanesulfonic acid, 3-amino-2-naphthoic acid, and tripeptides
(glutathione, Ser Ser Arg, Asp Asn Lys, and Arg Gln Arg) were detected
in all syrups. Some of these compounds may originally be presented in
the sap or generated via the chemical reactions during the thermal
treatment. Moreover, some polyphenols including medicagol, cis-
miyabenol C, and euphomine were also identified, which possibly
originated from the natural polyphenols in the sap and Kiam
(C. lanceolatum Craib) wood. Prasad et al. [61] found 3 major phenolic
compounds from nipa fruit including chlorogenic acid, protocatechuic
acid, and kaempferol. Some fragrance agents namely decyl isobutyrate
and 3-(L-menthoxy)-2-methylpropane-1,2-diol, the class of mono­
terpenoids, were surprisingly found in the nipa palm syrups. These
compounds may be a part of essential oil found originally in sap and
Kiam wood. Meconic acid (organic acid) and pentyl heptanoate (ester)
were also identified in nipa palm syrups, which may take part in the
acidity of the syrup. The presences of phytochemicals in the nipa palm
syrup may relate to the health benefits and the flavor characteristics (e.
g. floral and caramel aroma) of the syrup. Phetrit et al. [9] reported that
nipa palm syrup exhibited DPPH●-scavenging activity (25.7 μmol Tro­
lox equivalent (TE)/g, dry wt), ferric reducing antioxidant power (39.3
μmol TE/g, dry wt), and ferrous ion-chelating ability (0.4 μmol EDTA
equivalent/g, dry wt).
4. Conclusion
This study revealed the variations in physicochemical properties and
chemical compositions of nipa palm syrup harvested from Southern
Thailand. Results demonstrated that the physicochemical (e.g. color,
browning indices, HMF, viscosity, salinity, and acidity) and chemical
compositions (e.g. proximate composition, sugar profile, TPC, TFC,
minerals, non-protein nitrogenous compounds, phenolic compounds,
organic acids, and some flavor substances) of the nipa palm syrups
varied considerably within the plantation and the heating time.
Declaration of Competing Interest
The authors would like to declare that they do not have any conflict
of interest.
Acknowledgements
This work was funded by Ministry of Higher Education, Science,
Research and Innovation, Thailand. This research was financially sup­
ported by the new strategic research project (P2P), Walailak University,
Thailand.
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