original scientific paper

ISSN 1330-9862
https://doi.org/10.17113/ftb.60.02.22.7419

Nutritional and Functional Potential of Carob Syrup Versus Date

and Maple Syrups

Imad Toufeili1* , Marwa SUMMARY

Itani1 , Mona Zeidan1 , Research background. The carob tree (Ceratonia siliqua L.) is grown primarily for its
Osama Al Yamani2 and seeds that are utilized in the production of the highly prized locust bean gum. The mate-
rial left after the separation of seeds from the pods is utilized in the production of a range
Samer Kharroubi1
of traditional products including carob syrup, usually in cottage-type industries. The in-
ternational market penetration of carob syrups is rather limited and, accordingly, scant
Department of Nutrition and Food
1
information exists on their composition and phytochemical properties compared to main-
Sciences, American University of
Beirut, Riad El Solh 1107 2020, Beirut, stream syrups. The present study aims to determine key chemical parameters, phenolic
Lebanon profiles and antioxidant properties of carob syrups and benchmark these against those
Toxicology Research and Training
2
of date and maple syrups.
Center, Faculty of Health Sciences, Experimental approach. Carob syrups were prepared from 19 accessions of the carob,
American University of Science and
under laboratory conditions, by a similar procedure to those practiced by small-scale pro-
Technology, Beirut, Lebanon
ducers. The pH, browning index, the content of proteins, minerals, hydroxymethylfurfural,
Received: 9 July 2021 sugar composition, total phenols, antioxidant capacity and phenolic profiles of the pro-
Accepted: 8 February 2022 duced syrups along with branded samples of date and maple syrups were analyzed.
Results and conclusions. The pH and sugar composition of the carob syrups were com-
parable to those of date and maple syrups. In general, the carob syrups contained more
proteins, minerals, phenolic acids, flavonoids and total phenols, and exhibited higher an-
tioxidant capacity than the date and maple syrups. The carob syrups exhibited excessive
browning and contained more, or comparable content of hydroxymethylfurfural, than the
date and maple syrups. The data indicate that carob syrups provide more nutrients and
possess superior antioxidant potential to date and maple syrups. The high contents of the
carcinogenic hydroxymethylfurfural of the carob syrups warrant milder heating regimens
in the concentration step during production.
Novelty and scientific contribution. In contrast to studies based on commercial and/or
homemade syrups, this work utilized a relatively large number of laboratory-prepared
samples for creating a robust database for carob syrup. The results indicated that carob
syrups possess superior health promotion and disease prevention effects than the wide-
ly traded date and maple syrups. In addition to their potential positive contribution to
public health, carob syrups have been shown to be promising candidates for bolstering
the economic returns of farmers in carob-producing countries.
Keywords: syrup; Ceratonia siliqua L.; nutritional content; phenolic profile; antioxidant ca-
pacity; hydroxymethylfurfural

INTRODUCTION
Fruit- and tree sap-based syrups have been used for millennia as sweeteners in local
cuisines worldwide. In addition to providing sweetness, fruit and tree sap syrups contain
proteins, minerals, vitamins and a range of phytochemicals possessing antioxidant activ-
ity (1). Because of their superior health properties, food product developers are increas-
*Corresponding author:
ingly using these syrups as sugar substitutes to satisfy the demands of the health-con-
Phone: +96101343002
Fax: +96101744460 scious consumers for safer and more natural foods (2). Tree sap syrups are made by tapping
E-mail: toufeili@aub.edu.lb the trunks of endemic trees, collecting the exuding sap and concentrating the sap into a

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 Food Technol. Biotechnol. 60 (2) 266–278 (2022)

thick syrup (3). Fruit syrups are usually made by heating fruit against known/recognized commodities within the product
juices to different levels of total solids until the target consist- category are pivotal for their valorization, including possible
encies are attained (2). The heating step during the making recognition as Protected Designation of Origin (PDO) (16), and
of syrups increases viscosity, generates the brown colour and positioning in the global food market. Within this framework,
develops the unique flavour profiles of the products (4). In carob syrups were prepared, under laboratory conditions,
addition to their functionality as sweeteners, syrups are used from 19 carob accessions indigenous to Lebanon, and their
in the food industry to add viscosity, impart brown colour and physicochemical parameters, antioxidant capacity and phe-
desirable flavours, and mask bitterness in a range of food nolic profiles were determined and compared to those of
products (4). Amongst the tree sap syrups, maple syrup is the commercial maple and date syrups.
leading one with a forecasted global market value of $1.7 bil-
lion in 2023 (5). Data on the market value of fruit syrups are MATERIALS AND METHODS
difficult to locate. However, inferences about the size/market
penetration of fruit syrups can be made from the production Materials
statistics of the fruit, established practices of syrup produc- Potassium hexacyanoferrate(III) trihydrate (Carrez I), zinc
tion and the availability of literature on the properties and acetate dihydrate (Carrez II reagent kit), ammonium molyb-
uses of the syrup. To this end, the annual production of dates date, ammonium trioxovanadate(V), acetonitrile, water, so­
has been reported at 8.9 million tonnes in 2018 (6), with syrup dium metabisulfite, sodium bisulfite, sodium hydroxide, glu-
production being routinely practiced in date-growing coun- cose, fructose, sucrose, Folin-Ciocalteu reagent, iron(III)
tries (7) and frequently used as a sugar substitute in the for- chloride (hexahydrate), TPTZ (2,4,6-tri(2-pyridyl)-s-triazine),
mulation of foods (2,5). sodium acetate trihydrate, acetic acid, Trolox (6-hydro­-
Maple syrup exhibits interesting health properties includ- xy-2,5,7,8-tetramethylchromane-2-carboxylic acid), ABTS
ing more favourable metabolic responses than those gener- (2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), DPPH
ated after ingestion of refined sugar (8) and antioxidant, an- (2,2-diphenyl-1-picrylhydrazyl), potassium persulfate, gallic
ticancer and antimicrobial activities (9). Broadly similar health acid, p-coumaric acid, caffeic acid, t-cinnamic acid, syringic
properties have been ascribed to date syrup with reported acid, catechin, epigallocatechin-3-gallate, quercetin and the
antioxidant and antimicrobial activities (10). atomic absorption standards (Ca, Na, Mg and K) were ob-
In the Middle East and North Africa, the carob tree (Cera- tained from Sigma-Aldrich, Merck (Gillingham, Dorset, UK).
tonia siliqua L.) is widely cultivated due to its adaptability to Sodium carbonate, iron(II) sulfate, sodium thiosulfate and
harsh environmental conditions and the ability to grow on mercuric oxide were purchased from Merck KGaA (Darmstadt,
Germany), hydrochloric acid (37 %), sulfuric acid (95 %) and
marginally productive lands with low to medium rainfall
methanol were procured from VWR International (Lutter-
(250–500 mm/year) (11). The carob tree bears pod-shaped
worth, Leicestershire, UK). Light (amber colour grade A; Kirk-
fruits made up of a fleshy pulp that envelops several seeds.
land Signature, USA) and dark (dark colour grade A; Member’s
The seeds are rich in galactomannans and are commercially
Mark, USA) maple syrups, and date syrup (Alwadi Al Akhdar;
utilized in the production of locust bean gum. The carob pulp
Beirut, Lebanon) were procured from the local market.
contains appreciable amounts of sugars (chiefly sucrose, glu-
Nineteen carob accessions, growing in the different re-
cose and fructose), dietary fibre and polyphenols, and also
gions of Lebanon, were used in the preparation of carob syr-
some proteins and a range of minerals and vitamins (11). It is
ups. The accessions grew under diverse climatic conditions
utilized in the preparation of a range of traditional foods in-
ranging in elevation between 16 and 654 m, precipitation
cluding carob syrup, which is extensively used as a sweeten-
between 491 and 1038 mm, and average temperatures be-
er in many parts of the world (12). In addition to its sweeten-
tween 17 and 22 °C (17). The samples were sorted by remov-
ing functionality, carob syrup has been shown to possess
ing damaged pods and then washed with distilled water to
antioxidant activity (13) and superior anti-inflammatory and
remove adhering impurities. The pods were left to dry at
antimutagenic activities to cane, grape and sorghum syrups
room temperature (~25 °C) and were then placed in cloth
(14). Despite its long history of use as a sweetener and its po-
bags and stored at room temperature until use.
tentially valuable health-promoting properties, scant data
are available on the physicochemical and radical-scavenging
properties of carob syrup. Recently, the physicochemical Morphological parameters of carob pods
properties of homemade carob syrup have been reported The pod length (cm), width (cm), thickness (cm), mass (g)
(15). However, there is a dearth of information on the physico­ and the number of seeds/pod were measured on 10 random-
chemical properties of syrups prepared from different carob ly selected pods as described by Naghmouchi et al. (18) and
accessions under laboratory conditions. Still, no studies have are presented in Table S1.
attempted to compare, under the same test conditions, the
antioxidant potential of carob syrup to leading tree sap and
fruit syrups, viz. maple and date syrups. Creating a database Preparation of carob syrups
on the physicochemical parameters and potential health ef- The syrups were prepared according to commercial prac-
fects of traditional food products and benchmarking them tices followed in the production of carob syrup. The carob

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I. TOUFEILI et al.: Chemical and Antioxidant Profiles of Carob Syrup

pods were deseeded and the coarsely ground kibbles were The browning index (BI) was determined by measuring
soaked in distilled water (1:3 m/V) at room temperature for the absorbance of appropriate dilutions of the samples at 420
24 h and the resulting liquor was passed through a cheese- nm and converting it to the absorbance of the original sam-
cloth to remove suspended materials and then boiled in a ple (22).
steam-jacketed kettle until total solids of ~78–80 g/100 g
were reached. The hot syrups were placed in glass jars, cooled
Determination of minerals
promptly in running water (Fig. S1) and stored at 4 °C until
For the determination of Na, Mg, Ca and K, the syrup (~0.5
use.
g) was treated with concentrated HCl (15 mL) and heated at
200 °C for 30 min in a microwave digestion system (Ethos Up;
Chemical and physicochemical analyses Milestone, Sorisole, Italy). After cooling to room temperature,
Spectrophotometric analyses were performed with an the digest was diluted to 50 mL with deionized water, and Na,
Evolution 300 UV-VIS spectrophotometer (Thermo Fisher Sci- Mg, Ca and K were measured by atomic absorption spectros-
entific, Loughborough, UK) using Suprasil quartz cuvettes copy (Solaar S4 with ASX-510 autosampler; Thermo Fisher Sci-
(Mettler-Toledo Ltd., Leicester, UK). All analyses were per- entific) and the concentration was determined using stand-
formed in triplicate and results were reported on a dry mass ard curves prepared according to AOAC method 984.27 (23).
basis. P was measured colorimetrically by dry ashing the syrup (~2
g) at 550 °C for 16 h, dissolving the ash in concentrated HCl (2
mL) and heating to dryness. The resulting residue was dis-
Determination of total soluble solids, pH, moisture,
solved by heating in distilled water (10 mL) and the solution
protein, 5-hydroxymethylfurfural and browning index
was filtered and made up to 50 mL with distilled water. Ali-
Total soluble solids (TSS), pH, and 5-hydroxymethylfur- quots (5 mL) of the solution were treated with concentrated
fural (HMF) were determined according to the International HCl and ammonium molybdate-ammonium metavanadate.
Honey Commission (19). The TSS were determined at room The absorbance of the resulting yellow-coloured solution
temperature by placing enough syrup to evenly cover the was measured at 400 nm and the concentration was deter-
prism of an Abbe refractometer (Bellingham + Stanley, Kent, mined using a standard curve prepared with known concen-
UK) and reading the TSS in ˚Bx which represents mass fraction trations of phosphorus (0–50 µg/mL) (24).
of sucrose (1 ˚Bx=1 g sucrose per 100 g solution). The pH val-
ues of the syrups were determined at room temperature in
Determination of sugars
solutions of the samples (3 g in 15 mL of distilled water) with
Mass fractions of glucose, fructose and sucrose in the syr-
a pH meter (SevenCompact PH/Ion meter S220; Mettler-Tole-
ups were determined according to Fidan et al. (25) with some
do AG, Schwerzenbach, Switzerland). The mass fraction of
modifications. Syrup samples (~1 g) were dispersed in deion-
HMF was determined colorimetrically as per the procedure
ized water (25 mL), sonicated at 30 °C for 30 min and then fil-
of White by treating a solution of the syrup (2.5 g in 15 mL of
tered and stored at –18 °C until analysis. The mass fractions
distilled water) with both Carrez clarification reagent kit, Car-
of sucrose, fructose and glucose in the extracts were meas-
rez I (0.25 mL) and Carrez II (0.25 mL) solutions and making
ured with HPLC (LC-10A; Shimadzu, Kyoto, Japan) using a
up to 25 mL with distilled water (19). The solutions were fil-
Telos NH2 column (5 µm, 25 cm×4.6 mm; Kinesis Scientific Ex-
tered and aliquots of the filtrate (2 mL) were treated with wa-
perts, Redland Bay, Australia), Telos NH2 guard column (5 µm,
ter (2 mL) or 0.2 % sodium metabisulfite solution (2 mL), and
1 cm×4.6 mm), refractive index detector, and φ(acetonitrile,
the absorbance was read at 284 and 336 nm. The HMF mass
water)=70 % as mobile phase. The quantification of the sug-
fractions in the syrups were expressed in mg/kg.
ars was made using calibration curves constructed with
Moisture and protein (N×6.25) were determined accord- standard solutions of sucrose, glucose and fructose.
ing to AOAC methods 925.45 (20) and 955.04 (21), respective-
ly. Moisture was determined by mixing a diluted sample of
the syrup (1.2–1.5 g in ~10 mL water) with acid-washed sand, Determination of total phenolic content
heating on a steam bath (Labotec 402; Cape Town, South Af- The phenols were extracted by shaking the syrup (4 g)
rica) for 20–30 min, and then at 100 °C to a constant mass with φ(methanol, water)=50 % (10 mL) in a shaking water bath
(~3–4 h). Proteins were determined by treating the syrup (~2 (GFL Shaking Water Bath 1092; Gesellschaft fϋr Labortechnik
g) with HgO (0.7 g), anhydrous Na2SO4 (15 g) and concentrat- mbH, Burgwedel, Germany) for 30 min (26). The solution was
ed H2SO4 (25 mL) and boiling until a clear green liquid was filtered and the filtrate was kept at –80 °C until use.
obtained (~1.5 h). After cooling to room temperature, the The total phenolic content (TPC) was determined accord-
contents of the flask were diluted with distilled water (200 ing to Singleton et al. (27). An aliquot of the extract (1 mL) was
mL), treated with Na2S2O3 (25 mL, 0.3 M) and layered with con- mixed with 5 mL φ(Folin-Ciocalteu reagent, water)=50 % and
centrated NaOH (35 mL, 11 M). The NH3 in the flask was dis- 20 % Na2CO3 (4 mL), vortexed, incubated at room temperature
tilled into 0.1 M HCl and the excess HCl was titrated with 0.1 in the dark for 1 h and the absorbance of the solution was
M NaOH. measured at 765 nm. TPC was expressed in mg gallic acid

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 Food Technol. Biotechnol. 60 (2) 266–278 (2022)

equivalents (GAE) per 100 g syrup and concentration was de- composed of electrospray ionization (ESI) MS/MS ABI 4000
termined using a standard curve prepared with known con- Sciex® (Toronto, Canada) coupled with liquid chromatogra-
centrations of gallic acid (0–200 mg/L). phy station comprising a quaternary pump (LC-20AD-LPG-20),
autosampler (SIL-20A), column oven (CTO-20AC) and Vario
Antioxidant capacity determination by ferric reducing preparative octadecyl silica (VP-ODS) column (150 mm×2 mm
antioxidant power i.d., 5 µm) (Shimadzu®, Kyoto, Japan). The mobile phase com-
The ferric reducing antioxidant power (FRAP) assay was prised A: deionized water with 0.1 % acetic acid, and B: 0.1 %
carried out according to Benzie and Strain (28) with slight acetic acid in acetonitrile, and the gradient program was 0−4
modifications. An aliquot of the extract (100 µL) was mixed min 85 % A and 15 % B, 4.01−5.00 min from 15 % B to 50 % B,
with distilled water (900 µL), FRAP reagent (2 mL) and incu- 5.01−8.00 min 50 % A and 50 % B, and finally 8.01−12.00 min
bated at 37 °C for 30 min. The absorbance was measured at 85 % A and 15 % B. The flow rate of the mobile phase was 0.2
593 nm against a blank (1 mL water with 2 mL FRAP reagent). mL/min, the injection volume was 10 µL and the column tem-
FRAP was expressed in µM Fe(II) per 100 g syrup and concen- perature was set at 30 °C. The ESI parameters were 206 kPa
tration was determined using a calibration curve constructed for the nebulizer gas pressure, 172.4 kPa for the drying gas
with aqueous solutions of Fe(II) sulfate (0−100 µM). pressure and 400 °C for the ion source temperature. Stand-
ards of caffeic, t-cinnamic, p-coumaric, gallic and syringic
Antioxidant capacity determined by Trolox equivalent acid, catechin, epigallocatechin gallate and quercetin were
antioxidant capacity used in the quantification of phenols in the samples. Apart
The Trolox equivalent antioxidant capacity (TEAC) assay from catechin and syringic acid, which were analyzed in the
was performed as described by Fu et al. (29) with slight mod- negative mode with a needle voltage of –4500 V, data for the
ifications. An aliquot of the extract (100 µL) was added to the phenols were acquired in the positive mode and a needle
ABTS+• solution (3.8 mL) and kept in the dark at room temper- voltage of +5500 V.
ature for 30 min. The absorbance was measured at 734 nm
against a blank containing methanol, and TEAC was ex- Statistical analysis
pressed in mmol Trolox equivalents (TE) per 100 g syrup. Con-
Descriptive statistics was performed and presented to
centration was determined using a standard curve prepared
summarize the study variables of interest as mean values and
with known concentrations of Trolox (25−500 µM).
standard deviations. Values of the measured parameters were
subjected to one-way analysis of variance (ANOVA) and the
Antioxidant capacity determined by the DPPH method mean values were separated by Duncan’s multiple range test
The DPPH assay was performed as per Dhaouadi et al (13). when F-values were significant. All reported p-values were
An aliquot of the syrup extract (or a dilution therefrom) or based on two-sided tests and were compared with a signifi-
ascorbic acid standard solution (50 µL) was added at different cance level of 5 %. The Statistical Package for Social Sciences
concentrations (0–600 mg/L) to 60 µM DPPH in methanol (SPSS) v. 25.0 for Windows (30) was used to analyze the data.
(1950 µL) and incubated in the dark at room temperature for
30 min. The absorbance was measured at 515 nm against a
blank containing the same amount of DPPH˙ solution and 50 RESULTS AND DISCUSSION
µL of distilled water. The DPPH˙ inhibition (in %) was calculat-
Physicochemical parameters, browning index, protein
ed using the following equation:
and HMF mass fractions of carob, date and maple syrups
 ( A − As ) 
DPPH ⋅ inhibition =  0  ⋅100 /1/ While 89.7 % of the carob syrups had higher soluble sol-
 A0
  ids (p<0.05) than the date syrup, all carob syrups had higher
where A0 is the absorbance of the blank sample and As is the TSS (p<0.05) than the maple syrup (Table 1). Further, the TSS
absorbance of the tested solution (syrup extract or ascorbic of carob syrups prepared in the present work were higher
acid solution). than those of commercial syrups from Tunisia and Turkey,
The results were expressed as the extract concentration which are marketed at 73−75 and 66.6−73.7 g/100 g, respec-
providing 50 % inhibition (IC50) in mg extract per L as deter- tively (15) and those of date and maple syrup reported at 75
mined from the plot of the absorbance vs extract concentra- (7) and 67.1–67.4 g/100 g (9), respectively. In addition to being
tion. The IC50 values of the syrups were also compared to the set by national standards, the TSS contents are the chief de-
IC50 of known ascorbic acid solutions determined under the terminants of fruit and tree sap syrup viscosity/thickness
same conditions. which, expectedly, reflects broad national preferences.
All carob syrups had higher and lower pH (p<0.05) than
Identification and quantification of phenolic composition date and maple syrups, respectively (Table 1). The pH of fruit
by LC-MS/MS analysis and tree sap syrups depends on the content of organic acids
Individual phenols were quantified with liquid chroma- and minerals in the sap/juice, as well as microbial contamina-
tography-tandem mass spectrometry (LC-MS/MS) system tion during processing and storage (7). The pH values of date

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Table 1. Total soluble solids (TSS), proteins, browning index, pH and hydroxymethylfurfural (HMF) of carob, maple and date syrups
Carob accession w(TSS)/(g/100 g) w(protein)/(g/100 g) Browning index pH w(HMF)/(mg/kg)
Akkar (75.3±0.8)b (2.7±0.0)h (73.1±3.1)ef (4.9±0.0)jkl (1712.1±0.3)gh
Selaata (79.8±0.3)hi (2.50±0.01)g (68.3±1.1)de (4.93±0.02)kl (13515±426fg
Wadi El Hojeir (77.0±2.6)cd (1.83±0.02)e (87.8±0.2)g (4.72±0.01)h (1813±186)h
Batroun (78.3±0.5)efg (1.91±0.02)e (45.8±2.1)c (4.97±0.01)kl (881±94)cdef
Maaroub (78.7±0.4)fgh (1.85±0.03)e (31.3±1.0)b (4.93±0.08)kl (610±242)bc
Bourjin (78.5±0.3)fg (1.75±0.01)f (88.9±1.8)g (4.83±0.02)ij (1778±265)gh
Marjayoun (76.9±0.1)cd (1.77±0.00)e (49.0±1.1)c (5.17±0.04)m (779±6)bcd
AUB1 (77.6±0.1)def (2.14±0.01)f (49.1±1.4)c (4.68±0.01)gh (1271±303)ef
AUB2 (78.40±0.30)fg (4.40±0.06)k (85.5±10.5)fg (4.88±0.02)jk (1046±27)cdef
AUB3 (80.18±0.08)i (1.81±0.02)e (56.7±8.7)cd (4.90±0.13)jkl (814±117)bcde
AUB4 (79.43±0.06)ghi (3.2±0.3)j (57.9±1.2)g (4.45±0.01)d (519±210)h
AUB6 (80.3±0.2)i (2.39±0.06)g (49.8±2.1)c (4.75±0.05)hi (874±50)bcde
AUB7 (76.3±0.3)bc (2.94±0.07)i (52.4±4.0)c (4.56±0.00)ef (1103±352)def
AUB8 (78.8±0.2)gh (0.98±0.02)b (32.4±3.3)b (5.0±0.1)l (711±91)bcd
AUB9 (78.5±0.4)fg (4.82±0.01)l (86.9±12.1)cd (4.37±0.01)c (2169±514)b
AUB10 (78.9±0.1)gh (1.35±0.02)c (88.1±0.6)g (4.62±0.04)fg (1784±172)gh
AUB11 (78.8±0.2)gh (2.5±0.1)g (32.1±6.9)b (4.73±0.02)h (1121±302)def
AUB12 (80.3±0.3)i (2.22±0.01)f (109.9±16.6)h (4.25±0.00)b (4049±182)i
AUB14 (77.25±0.05)cde (2.97±0.02)i (127.3±5.4)i (4.49±0.01)de (1810±250)h
Date syrup (75.50±0.00)b (1.55±0.07)d (98.0±23.4)gh (4.01±0.03)a (1987±283)h
Amber maple syrup (66.6±0.1)a (0.08±0.01)a (3.9±0.2)a (6.00±0.06)n (66.2±3.0)a
Dark maple syrup (66.9±0.1)a (0.16±0.00)a (4.1±0.2)a (6.60±0.06)o (72.5±2.2)a
Range of carob syrups 75.3−80.3 1.0−4.8 31.3−127.3 4.3−5.2 520−4049
Mean±S.D. of carob syrups 78.4±1.4 2.5±0.9 67.2±27.2 4.7±0.2 1379±809
All mass fractions were determined on dry mass basis. AUB1 to AUB14=American University of Beirut. Values with different letters in superscript
in the same column are significantly different (p<0.05) according to Duncan’s multiple range test

and maple syrups were close to those reported at 4.2 (31) and (Table 2) and mild conditions for concentrating the maple sap
6.7−7.1 (9), respectively. Furthermore, the pH of carob syrup including heating conditions needed to develop the typical
samples was similar to those of samples marketed in Tunisia colour and flavour of the finished product (34,35). The lower
and Turkey with ranges of 4.4–5.4 (15). degree of browning of the carob syrup than of date syrup
All carob syrups contained more proteins (p<0.05) than might have resulted from their lower mass fractions of reduc-
maple syrup, while 89.7 % of the carob syrups had higher pro- ing sugars (p<0.05) (Table 2) and higher pH (p<0.05), which
tein mass fractions (p<0.05) than the date syrup sample (Ta- limits the inversion of sucrose during the heat concentration
ble 1). These findings indicate that carob syrup provides more step.
proteins for human nutrition than date and maple syrups. The The maple and date syrup samples had the lowest and
protein mass fraction of the date syrup sample was close to highest average levels of HMF, respectively. Furthermore,
that of commercial samples at 1.14−1.45 g/100 g (32), while while only one sample did not differ in HMF content (p>0.05),
the maple syrup samples contained fewer proteins than re- the other 18 carob syrups contained more HMF (p<0.05) than
ported at 0.37 g/100 g (33). the maple syrup samples. Only one sample contained more
The maple syrup samples browned the least (p<0.05), as HMF (p<0.05), while the other carob syrup samples either did
measured by the BI, when compared to the other syrups (Ta- not show differences (p>0.05) or contained less HMF than the
ble 1). Furthermore, the carob syrups browned less than date date syrup sample (Table 1). Hydroxymethylfurfural is formed
syrup, as reflected by the lower BI (p<0.05) of 89.5 % samples through the dehydration reactions that take place during the
and the BI values reported for 6 homemade carob syrup sam- caramelization of sugars and their heating in the presence of
ples from Tunisia at a mean BI value of 34 (15) (Table 1). The amino compounds in the Maillard reaction (34). While the for-
BI reflects the content of melanoidins (34) produced through mation of HMF during caramelization of sugars is independ-
the Maillard reaction between sugars and amino compounds ent of the confounding effects of amino compounds, the de-
and caramelization of sugars upon heating and subsequent termination of the precise contribution of caramelization and
storage. The low degree of browning observed in the maple the Maillard reaction to the formation of HMF in heated syrup
syrup samples, as compared to the other syrups, could be at- containing sugar and amino compounds is a daunting task;
tributed to their markedly lower protein content (p<0.05), however, during heating of model systems containing fruc-
higher pH (p<0.05), lower content of reducing sugars (p<0.05) tose and lysine, 10−36 % of the produced HMF derives from

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the caramelization reactions (36). The formation of HMF dur- Table 2. Fructose, glucose and sucrose contents on dry mass basis of
ing caramelization and heating of syrup is influenced primar- carob, maple and date syrups
ily by the types and concentrations of sugars and amino com- w/(g/100 g)
Carob accession
pounds, heating regimen and pH. To this end, the HMF mass Fructose Glucose Sucrose
fractions tend to be higher in the systems comprising reduc- Akkar (12.1±1.0)b (7.0±0.8)bc (62.0±1.7)hi
Selaata (17.7±1.0)cd (5.9±0.8)b (59.3±3.0)gh
ing sugars, basic amino acids and acidic pH (37). The low HMF
Wadi El Hojeir (21.7±1.4)fgh (10.7±0.9)gh (59.3±3.0)gh
mass fractions in the maple syrup samples can be attributed
Batroun (18.8±2.5)de (10.0±1.5)fgh (74.4±9.8)k
to their lower contents of glucose, fructose (p<0.05) and pro-
Maaroub (16.8±0.7)cd (8.3±0.6)cdef (65.2±3.8)hij
teins (p<0.05) and higher pH (p<0.05) than of the carob syr-
Bourjin (13.0±1.2)b (7.1±1.0)bcd (61.9±6.0)hi
ups (Table 1 and Table 2), and the mild heating regimen ap-
Marjayoun (15.7±0.7)c (5.7±0.2)b (67.3±1.0)ijk
plied in their production (35). The higher HMF mass fractions
AUB1 (23.6±2.2)hi (12.7±1.5)i (53.7±4.9)fg
in the date syrup samples than in carob syrup samples might
AUB2 (17.8±1.5)cd (9.4±1.1)efgh (49.9±4.0)ef
have been caused by their higher mass fractions of glucose
AUB3 (19.3±2.2)def (10.8±1.3)gh (62.2±8.0)hij
and fructose (p<0.05) and lower pH (p<0.05) in view of the
AUB4 (19.1±0.5)j (10.6±2.0)k (39.7±1.1)d
similar concentration procedure, entailing open pan evapo- AUB6 (23.0±0.7)gh (13.2±0.8)i (43.4±2.2)de
ration, practiced in the production of carob and date syrups. AUB7 (25.4±1.7)i (16.9±1.0)j (58.1±4.0)gh
Pearson’s correlation analysis between HMF mass fractions AUB8 (16.2±1.1)c (8.0±0.5)cde (71.3±5.3)jk
and pH, sucrose and fructose of the syrups was –6.58 (p<0.01), AUB9 (39.8±0.9)def (25.6±0.4)gh (5.4±0.3)a
–0.647 (p<0.01) and 0.510 (p<0.05), respectively. These corre- AUB10 (20.8±1.6)efg (11.5±0.7)hi (42.7±3.2)de
lations are in accordance with previous findings from model AUB11 (19.2±1.2)def (8.9±1.2)defg (71.0±4.5)jk
systems where the formation of HMF was shown to be fa- AUB12 (21.1±2.6)efg (9.4±0.2)efgh (31.7±3.9)c
voured at acidic pH values due to presence of fructose (38). AUB14 (37.6±0.5)j (24.9±2.8)k (24.5±4.3)b
Furthermore, the negative correlation between sucrose and Date syrup (43.2±0.8)k (42.9±0.8)l (3.7±0.8)a
HMF mass fractions is indicative of the lower reactivity of the Amber maple syrup (0.34±0.00)a (0.00±0.00)a (95.0±1.7)l
non-reducing disaccharide sucrose in caramelization and the Dark maple syrup (0.35±0.00)a (0.35±0.00)a (96.5±4.2)l
Maillard-type browning, as reflected in the lower tendency Range of carob
12.1−39.8 5.7−25.6 5.4−74.4
of the high-sucrose containing syrups, and notably the maple samples
syrups, to undergo browning and accumulate HMF (Table 1 Mean±S.D. of carob
21.0±7.1 11.4±5.6 52.9±17.9
syrups
and Table 2).
Notwithstanding its beneficial antioxidant activity, HMF Values with different letters in superscript in the same column are
significantly different (p<0.05) according to Duncan’s multiple range
has been shown to be carcinogenic and mutagenic and to test. AUB1 to AUB14=American University of Beirut
induce hepato- and nephrotoxicity in laboratory animals,
thereby prompting regulatory agencies to set limits to its
(Table 2). The mass fractions of glucose, fructose and sucrose
contents in foods (38). Among different approaches to miti-
of the carob syrups included the mass fractions reported for
gate the contents of HMF in foods, reducing the severity of
10 commercial carob syrups from Turkey (41). The glucose,
heat treatment during the concentration step in the produc-
fructose and sucrose mass fractions of maple and date syrups
tion of fruit and tree sap syrups is the most effective. To this
were similar to those reported for commercial date (32) and
end, ~10- and 2.5-fold reduction in HMF mass fractions were
maple syrups (3). The high sugar mass fraction of carob syrup
achieved upon reducing the temperature during the sap/
juice concentration from 100 to 70 °C in the production of is responsible for its widespread utilization in the making of
date (39) and palm sugar syrups (40), respectively. The HMF a range of ethnic products and its potential use as sugar sub-
mass fractions of the carob syrup samples included the HMF stitute in the formulation of healthier confections in view of
values reported for Tunisian carob syrup at 450 (15) and the provision of nutrients and phytochemicals in addition to
1000−2675 mg/kg reported for commercial date syrup pro- sweetness. Furthermore, because of its high sugar content,
duced by open pan evaporation (39). carob syrup has been reported to be a promising substrate
for the production of a range of fine chemicals by industrial
fermentation (42).
Sugar composition of carob, date and maple syrups
All carob syrups contained more (p<0.05) glucose and
fructose and less sucrose (p<0.05) than maple syrup samples.
Mineral composition of carob, date and maple syrups
They also had less glucose and fructose (p<0.05) and, apart In general, the majority of carob syrups (13−19 samples;
from 1 sample that showed no differences (p>0.05), more su- 68.4−100 %) contained more K (p<0.05), P (p<0.05), Mg
crose (p<0.05) than date syrup (Table 2). The sugar contents (p<0.05) and Na (p<0.05), and less Ca (p<0.05) while the oth-
of the carob syrups appeared to be intermediate to those of ers either did not show differences (p>0.05) (3−6 samples;
maple syrup that contains almost only sucrose, and date syr- 16−32 %) or contained less (p<0.05) (1−2 samples; 5.3−10.5
up where an invert-sugar-like composition predominates %) minerals than the maple syrup samples (Table 3). Also, the

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Table 3. Mineral composition of carob, date and maple syrups

w/(mg/100 g)
Carob accession
P Ca Mg K Na
Akkar (116.5±3.0)a (234.28±0.00)k (83.14±4.00)h (1238±34hi (7.9±0.8)ab
Selaata (704±96)fgh (184.3±1.1)hi (46.1±0.8) )cde (885±37)de (30.5±1.9)c
Wadi El Hojeir (622±21)efg (170.8±21.4)gh (45.8±8.8) )cde (1073±20)fg (39.9±0.4)e
Batroun (536±6)cde (169.8±3.3)gh (59.8±3.0)g (1037±60)fg (43.2±5.5)e
Maaroub (551±10)def (149.1±15.0)fg (42.5±6.4)cd (945±44)de (31.9±5.8)c
Bourjin (606±44)efg (162.6±53.0)gh (41.5±3.9)cd (789±54)cd (40.1±4.9)e
Marjayoun (659±48)efgh (108.1±2.6)bcd (51.6±2.3)efg (1102±48)g (37.6±2.8)de
AUB1 (738±106)gh (120.9±12.4)cdef (58.9±4.6)g (975.2±8.2)ef (50.6±3.8)f
AUB2 (788±42)hi (97.4±19.9)abc (52.7±7.1)efg (846.8±4.7)cde (29.7±1.7)c
AUB3 (791±98)hi (111.5±1.8)bcde (52.2±3.7)efg (952±28)ef (41.6±4.6)e
AUB4 (108±27)i (129.0±15.4)bcd (40.3±6.1)def (77998)i (9.5±0.1)de
AUB6 (589±32)efg (113.6±11.8)bcde (48.1±2.7)def (970±136)ef (34.2±3.4)cd
AUB7 (258±32)ab (77.6±18.5)a (25.6±4.2)a (735±55.bc (12.4±1.4)b
AUB8 (631±25)efg (95.3±4.4)abc (37.8±2.1)bc (816±24)cd (49.8±2.3)f
AUB9 (908±174)a (110.8±6.9)def (48.2±1.7)bcd (1324±74)cd (37.8±2.6)ab
AUB10 (1714±400)j (112.5±0.6)bcde (33.2±0.0)ab (1148±39)gh (94.7±1.9)h
AUB11 (626±83)efg (142.4±18.6)efg (52.5±6.5)efg (852.4±4.6)cde (39.2±2.9)de
AUB12 (287±12)b (142.4±12.3)efg (55.7±8.4)fg (1536±68)j (29.6±2.9)c
AUB14 (409±12)bcd (86.8±7.4)ab (32.4±3.9)ab (631±43)b (79.7±3.3)g
Date syrup (561.3±2.8)ef (166.4±3.2)gh (78.4±0.9)h (1103±168)g (11.5±0.3)b
Amber maple syrup (392±22)bc (220.5±11.7)jk (33.4±0.4)ab (322.2±1.9)a (5.9±1.1)a
Dark maple syrup (120.46±6.08)a (203.1±22.9)ij (32.1±2.0)ab (350±19)a (5.2±0.4)a
Range of carob syrups 108−1714 77.6−234.3 25.6−83.1 631−1536 7.9−94.7
Mean±S.D. of carob syrups 613±348 132.6±39.0 47.8±12.5 977±222 38.4±21.0
Mass fractions are expressed on dry mass basis. Values with different letters in superscript in the same column are significantly different
(p<0.05) according to Duncan’s multiple range test. AUB1 to AUB14=American University of Beirut

majority of carob syrups (16−18 samples; 84.2−84.7 %) con- 8 samples from Tunisia (2.1−2.2 g/100 g) (15) and 10 samples
tained more Mg (p<0.05) and Na (p<0.05) and 1−4 samples from Turkey (0.72−1.2 g/100 g) (41). The TPC (expressed as GAE
(5.2−21.1 %) contained more (p<0.05) Ca, K and P, while the on dry mass basis) of maple syrup samples was higher than
others either did not show differences (p>0.05) (1−10 sam- reported for amber and dark maple syrups at (45.6±18.7) and
ples, 5.3−52.6 %) or contained less (p<0.05) (5−10 samples; (72.0±18.2) mg/100 g, respectively (9). The high TPC of carob
26.3−52.6 %) minerals than date syrup (Table 3). The relative syrups is an added advantage of the use of these syrups as
contents of the investigated minerals in maple syrup were sugar substitutes in the formulation of foods in view of their
similar to those reported for commercial maple syrups with ability to quench reactive oxygen species and, consequently,
the highest and lowest contents of K and Na, respectively (3). to retard/mitigate the development/harmful effects of de-
The mineral composition of the date syrup was, in general, generative diseases (43).
within the reported ranges (10) (Table 3). These findings indi- Apart from 5 samples that did not differ (p>0.05) from
cate that carob syrup is a significant source of K and a good maple syrup, all carob syrups had higher antioxidant capaci-
source of Ca, Na, Mg and P and will potentially be a significant ty (p<0.05), as determined by the FRAP assay, than maple syr-
contributor to the mineral nutrition of consumers. up (Table 4). Moreover, 14 carob syrup samples (73.7 %) did
not differ (p>0.05), while 5 samples (26.3 %) showed higher
Total phenols and antioxidant capacity antioxidant capacity (p<0.05), according to the FRAP assay,
Apart from 2−4 samples, which did not differ in their TPC than date syrup (Table 4).
(p>0.05), carob syrup samples had higher TPC (p<0.05) than All carob syrups showed higher antioxidant capacity
maple syrup samples (Table 4). Additionally, apart from 3 (p<0.05), as determined by the TEAC assay, than the maple
samples that had lower TPC (p<0.05), carob syrup samples syrup samples and, apart from 4 samples (21.1 %) that did not
had higher (p<0.05) (9 samples, 47.4 %) than or did not differ show differences (p>0.05), the carob syrup samples had high-
in their TPC (p>0.05) (7 samples, 36.8 %) from date syrup (Ta- er antioxidant capacity (p<0.05) than the date syrup (Table 4).
ble 4). Notwithstanding the different solvents used in the ex- All carob syrups showed higher antioxidant capacity
traction of polyphenols from the syrups, carob syrups includ- (p<0.05), as determined by the DPPH assay, than the maple
ed the average TPC (expressed as GAE on dry mass basis) of syrup samples. Also, 3 carob syrup samples had higher anti-

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 Food Technol. Biotechnol. 60 (2) 266–278 (2022)

oxidant capacity (p<0.05), while the others did not show dif- The antioxidant potential of foods is frequently deter-
ferences in their antioxidant capacity (p>0.05) from that of mined by a combination of several tests that are based on
the date syrup (Table 4). The carob syrups had higher mean different principles and expressed in different units. Accord-
antioxidant capacity (lower IC50) than that reported for carob ingly, indices that integrate the data from different antioxi-
syrup at 47.2 mg/L (13). For comparison, ascorbic acid showed dant assays are often utilized to construct measures of the
an IC50 of (74±5.7) mg/L under the same conditions in the total antioxidant capacity of foods (46). To this end, the data
present work. The IC50 values of maple syrup (647.1 and 501 from the different antioxidant tests were normalized and the
mg/L) in DPPH assay were higher than the IC50 values of their derived z-scores were averaged to generate relative antioxi-
ethanol extracts at 97.6−102.4 mg/L (44). dant capacity indices (RACI) (46) for the different syrups (Fig.
Notwithstanding the differences in the assay protocols 1). All the carob syrups exhibited higher RACI than maple syr-
up and, apart from 3 carob syrups that had close RACI, high-
and the use of extracts or whole syrups in the determinations,
er than date syrup (Fig. 1). The TPC, as determined by the Fo-
carob syrups have been reported to contain significant quan-
lin-Ciocalteu reagent, correlated strongly with the RACI
tities of total phenols and to exhibit high antioxidant capac-
(r=0.881, p<0.01), thereby offering further support to the
ity, comparable to that of butylated hydroxytoluene, in sev-
modulation of the antioxidant capacity of foods and biolog-
eral in vitro antioxidant capacity assays (13,15,41). Similar
ical materials by their endogenous phenolics (47).
findings have been reported for the total phenols and anti-
Given the pivotal role of phenols and their associated an-
oxidant capacity of maple (1,9,44) and date syrups (45). How- tioxidant potential in combating the development and pro-
ever, when analyzed under the same conditions, the vast ma- gression of degenerative diseases and mitigating their ill ef-
jority of carob syrup samples had higher TPC than amber and fects (43), the present findings indicate the superiority of
dark maple syrup and comparable or higher TPC than date carob syrup as a functional dietary ingredient as compared
syrup (Table 4). Similar patterns were observed for the anti- to date and maple syrups. The reported higher anti-inflam-
oxidant capacity with most of the carob syrup samples exhib- matory and antimutagenic effects of carob syrup than those
iting higher antioxidant capacity than amber and dark maple of cane, grape and sorghum molasses (14) further attest to its
syrup and comparable or higher than date syrup in the ABTS, advanced health-promoting effects relative to those of cere-
DPPH and FRAP assays (Table 4). al, fruit and tree sap syrups.

Table 4. Total phenols and antioxidant capacity of carob, maple and date syrups expressed on dry mass basis
w(total phenols as GAE)/ Antioxidant capacity
Carob accession
(g/100 g) FRAP/(μmol/100 g) IC50(g/L) TE/(mmol/100 g)
Akkar (728±84)f (2.0±0.0)efg (32.0±2.0)ab (5.6±1.1)ghi
Selaata (732±73)f (1.4±0.3)bcde (16.5±1.5)ab (6.3±0.8)hi
Wadi El Hojeir (1368±40)g (1.65±0.03)cdef (17.0±0.6)ab (5.6±0.6)ghi
Batroun (443±69)cd (0.67±0.06)ab (18.6±0.9)ab (4.2±0.7)def
Maaroub (370±84)bc (1.4±0.5)bcde (46.2±6.8)b (2.5±0.4)bc
Bourjin (781±56)f (2.4±1.0)fg (16.6±1.5)ab (8.3±0.9)j
Marjayoun (358±12)b (1.14±0.08)bcd (26.6±0.9)ab (3.2±0.8)bcd
AUB1 (494±15)de (1.13±0.04)bcd (14.4±1.9)ab (4.6±0.1)efg
AUB2 (701±62)f (2.68±0.04)g (13.7±0.8)ab (4.9±0.6)efg
AUB3 (521±13)de (0.94±0.08)bcd (18.7±0.0)ab (4.4±0.8)defg
AUB4 (442±48)f (1.4±0.1)bcde (34.1±0.9)ab (3.6±0.7)cde
AUB6 (453±8)de (1.5±0.2)bcde (24.2±1.8)ab (2.3±1.0)bc
AUB7 (487.4±1.3)e (1.8±0.3)def (21.1±2.8)ab (4.2±0.2)def
AUB8 (345±52)b (0.86±0.02)abc (36.3±0.7)ab (2.5±0.2)bc
AUB9 (734±84)cd (1.4±0.8)bcde (20.2±0.2)ab (5.2±0.5)fgh
AUB10 (1566.4±2.3)h (2.7±0.4)g (10.1±1.3)ab (6.6±0.7)i
AUB11 (336±38)ab (1.02±0.08)bcd (36.3±1.0)ab (2.4±0.2)bc
AUB12 (2215±6)i (14.9±1.4)h (4.9±0.7)a (20.5±2.2)k
AUB14 (741±17)f (2.4±0.8)fg (19.9±0.0)ab (5.6±0.5)ghi
Date syrup (461±13)de (1.06±0.02)bcd (33.7±0.6)ab (2.1±0.0)b
Amber maple syrup (261.0±2.6)a (0.10±0.00)a (647±14)c (0.4±0.0)a
Dark maple syrup (314.0±2.3)ab (0.11±0.00)a (501±59)d (0.4±0.0)a
Range of carob syrups 336−2214 0.7−14.9 4.9−46.2 2.3−20.5
Mean±S.D. of carob syrups 729±487 2.3±3.1 23.0±10.8 5.4±4.0
GAE=gallic acid equivalents, TE=Trolox equivalents, FRAP=ferric reducing antioxidant power, TEAC=Trolox equivalent antioxidant capacity,
DPPH=2,2-diphenyl-1-picrylhydrazyl. Values with different letters in superscript in the same column are significantly different (p<0.05)
according to Duncan’s multiple range test. AUB1 to AUB14=American University of Beirut

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I. TOUFEILI et al.: Chemical and Antioxidant Profiles of Carob Syrup

5 side of PC2, thereby suggesting that this principal compo-

carob syrups nent is mainly associated with antioxidant capacity of the syr-
4 date syrup
amber maple syrup ups given the high correlation of the TPC and antioxidant ca-
dark maple syrup
3 pacity with the reported antioxidant properties of HMF (38).
While the carob syrups showed a diffuse pattern in sugar
2
composition, the date and maple syrups loaded heavily on
RACI

1
the opposite sides of PC1 in congruence with the invert-sug-
ar-like composition of the former and the almost exclusive
0 presence of sucrose in the latter. All the carob syrups loaded
higher than the maple syrups on PC2, which is indicative of
-1
their higher TPC, HMF and antioxidant capacity levels; how-
-2 ever, the date syrup loaded amongst the carob syrups on PC2
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22
in accord with its TPC, HMF and antioxidant capacity levels
Syrup
being within the ranges of these parameters exhibited by the
Fig. 1. Relative antioxidant capacity indices (RACI) of the analyzed carob syrup samples.
syrups (carob syrups prepared from the carob accessions as listed in
Tables 1−5)
Phenolic profiles of carob, date and maple syrups
Most carob syrup samples contained more (p<0.05) phe-
The acquired data were subjected to principal compo- nolic acids (caffeic, t-cinnamic, p-coumaric, gallic and syring-
nent analysis (PCA) to discern the variables that are operative ic acids) with few showing no differences in their phenolic
in mediating the relationships amongst the syrups and to acid contents from amber and dark maple syrups (Table 5). In
identify possible groupings of the investigated samples. Two the analyzed flavonoids, all carob syrup samples contained
principal components accounting for 62.5 % of the total var- less catechin (p<0.05), while most did not show differences
iance separated the syrups into distinct groups (Fig. 2). The (7−15 samples, p>0.05) or contained more (4−12 samples,
first principal component (PC1), which explained 47.1 % of the p<0.05) quercetin than maple syrup (Table 5). Furthermore,
total variance, related chiefly to the sugars (glucose, fructose all carob syrup samples contained less (p<0.05) caffeic acid,
and sucrose), proteins, pH, TSS, BI and Ca, while the second more (p<0.05) p-coumaric, syringic and t-cinnamic acids, and,
principal component (PC2) was associated mainly with the BI, apart from 8 samples that did not show differences (p>0.05),
HMF, TP, RACI and K. It is noteworthy that the reactants, con- more gallic acid than date syrup (Table 5). Moreover, apart
ditions, the index of caramelization and the Maillard-type of from 7 and 8 samples that did not exhibit differences (p>0.05),
browning loaded on PC1. To this end, fructose, glucose, pro- carob syrup samples contained more (p<0.05) catechin and
teins and BI loaded on the positive side of PC1, thereby con- quercetin than date syrup (Table 5). Of the analyzed pheno-
firming the direct relationship between browning intensity lics, gallic acid correlated with TPC (r=0.864, p<0.01) and RACI
and these reactants, while the pH and sucrose loaded heavi- (r=0.972, p<0.01), quercetin with TPC (r=0.586, p<0.01) and
ly on the negative side of PC1 in accord with the intensifica- RACI (r=0.621, p<0.01) and catechin with RACI (r=–0.490,
tion of browning at acid pH values and the sluggish reactivi- p<0.05), thereby suggesting that these phenolics are the
ty of sucrose in browning under the conditions of syrup most operative in shaping the TPC and antioxidant capacity
production. The TP, RACI, HMF and BI loaded on the positive of the investigated syrups.
The phenolic profiles of fruit and tree sap syrups are
shaped by the liquid-liquid extraction protocol, use of resins
4
C18 or solid-phase extraction to remove interfering compounds,
3
and the chromatographic conditions employed in the iden-
2 tification and quantification of the individual phenolics. Ac-
1
C1

C3 RACI
TP HMF
cordingly, only broad comparisons of phenolic profiles of syr-
PC 2 (15.4 %)

ups reported by different authors are plausible. Under the

C16
BI
C6 Mg TSS
C2 D
Ca Pro Fru C15
PNa Glu
0 C4
C8 C9
Suc
pH
C7
C5
C17

C14
C10
C11
C12

C13
C19

analytical conditions used in this work, gallic acid was the

major phenolic in carob, date and maple syrups (Table 5). The
-1 MD MA

-2
preponderance of gallic acid in carob syrup has been attrib-
-3 uted to its preferential extraction and release to other gallic
-4
acid-containing phenolics in the kibbles during syrup prepa-
-4 -3 -2 -1 0 1 2 3 4
PC 1 (47.1 %) ration (48). Furthermore, notwithstanding the differences in
Fig. 2. Principal component analysis biplot showing relationships analytical protocols, the phenolic patterns of the carob syr-
amongst the chemical and physicochemical parameters and the ups obtained in the present work were, in general, compara-
analyzed syrups (C1–C19 refer to the carob syrups prepared from ble to those reported elsewhere (13). The predominance of
the carob accessions as listed in Tables 1−5). Fru=fructose, Glu=glu-
cose, Suc=sucrose, HMF=5-hydroxymethylfurfural, Pro=proteins,
gallic acid in the date syrup may be related to it being the
RACI=relative antioxidant capacity, TP=total phenols, D=date syrup, main phenol in different varieties and clones of dates (49). In
MA=maple syrup amber, MD=maple syrup dark contrast to the present findings, protocatechuic acid,

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Table 5. Mass fractions of phenolic acids and flavonoids of carob, maple and date syrups expressed on dry mass basis
w/(μg/g)
Carob accession t-cinnamic p-coumaric
Gallic acid Syringic acid Caffeic acid Catechin Quercetin
acid acid
Akkar (4266±6)cdef (1.2±0.3)f (0.9±0.2)bcdef (14.2±1.0)de (13.3±0.8)hi (1.0±0.1)de (1.21±0.01)abc
Selaata (770±46)1j (0.08±0.02)a (1.01±0.1)cdefg (17.6±2.7)gh (12.9±2.2)gh (1.05±0.03)de (1.31±0.05)cde
Wadi El Hojeir (1377±91)gh (1.93±0.01)hi (1.3±0.1)9h (15.0±0.8)ef (11.0±1.0)efgh nda (1.56±0.00)f
Batroun (204±19)ab (0.92±0.06)def (0.72±0.03)b (15.5±2.8)efg (9.8±1.1)cdef (1.28±0.03)de (1.3±0.2)cde
Maaroub (565±7)fg (1.67±0.05)gh (0.78±0.06)bc (13.7±0.5)de (8.8±0.5)cde (1.6±0.3)f (1.69±0.03)f
Bourjin (2351±333)l (0.41±0.08)bc (1.12±0.01)fgh (8.1±0.9)bc (15.34±0.02)ij (0.19±0.01)a (1.23±0.02)abcd
Marjayoun (523±15)efg (0.34±0.09)abc (1.04±0.01)defgh (8.4±0.2)bc (8.2±1.4)bcd (0.63±0.08)bc (1.23±0.01)abcd
AUB1 (828±76)hi (0.90±0.06)de nda (10.4±1.2)c (13.3±0.8)hi (1.0±0.1)de (1.32±0.03)cde
AUB2 (955±67)i (1.75±0.30)ghi nda (7.7±0.5)b (10.62±0.07)efg (0.31±0.07)ab (1.29±0.04)bcde
AUB3 (657±84)bcdef (0.63±0.02)cd (1.1±0.1)fgh (8.1±1.7)bc (15.9±1.2)j (0.88±0.08)cde (1.19±0.06)ab
AUB4 (394±24)hi ndj (0.8±0.1)defgh (18.3±0.8)fgh (25.4±0.1)def (0.07±0.02)a (1.25±0.02)abcd
AUB6 (489±61)cdefg (0.8±0.2)de (0.89±0.09)bcdef (23.5±0.4)j (11.8±1.2)fgh (1.0±0.1)de (1.55±0.04)f
AUB7 (659±21)gh (1.6±0.5)g (0.77±0.09)bc (19.88±0.01)i (12.9±1.3)gh (1.1±0.2)de (1.35±0.06)e
AUB8 (361±24)abcd (2.0±0.3)ij (1.06±0.09)efgh (29.0±2.1)k (15.7±0.8)j (1.0±0.1)de (1.17±0.05)ab
AUB9 (838±27)bcde (2.3±0.2)a (1.0±0.1)bcd (16.6±1.5)hi (9.9±1.4)l nda (1.23±0.02)abcde
AUB10 (1891±186)k (1.1±0.2)ef (0.8±0.1)bcde (12.6±0.9)d (7.6±0.8)bc (1.2±0.2)e (1.31±0.02)cde
AUB11 (293±70)hi (0.16±0.07)ab (0.77±0.02)bc (22.6±2.5)j (17.6±1.8)jk (0.8±0.2)cd (1.15±0.07)a
AUB12 (7666±370)m (2.02±0.07)ij (0.89±0.01)bcdef (8.8±0.6)bc (6.0±1.1)b nda (1.84±0.08)g
AUB14 (948±30)i (1.6±0.2)gh (1.2±0.2)gh (15.0±1.3)ef (18.5±3.7)k (0.33±0.07)ab (1.33±0.08)de
Date syrup (254±17)abc nda (7.7±0.4)i nda (2.8±0.2)a (0.14±0.02)a (1.15±0.03)a
Amber maple syrup (135±26)a nda nda nda (0.71±0.08)a (2.6±0.3)g (1.23±0.02)abcd
Dark maple syrup (221±86)abc nda nda nda (0.9±0.1)a (1.75±0.03)f (1.25±0.03)abcde
Range of carob syrups 204–7666 nd-2.26 nd-1.26 7.7–29.0 6.0–25.4 nd-1.57 1.2–1.8
Mean±S.D. of carob syrups 1166±1666 1.1±0.7 0.8±0.3 15.0±5.9 12.9±4.6 0.7±0.5 1.3±0.2
Values with different letters in superscript in the same column are significantly different (p<0.05) according to Duncan’s multiple range test.
dm=dry mass, nd=not detected. AUB1 to AUB14=American University of Beirut

coniferyl alcohol and vanillin were the major phenolics in a favourable phenolic profile, provides more proteins and
methanol extracts of maple syrup prepared by solid-phase minerals, contains more total phenols and has a higher anti-
extraction (50). oxidant potential than maple and date syrups. These traits
render carob syrups strong candidates for the category of
mainstream syrups in international trade with obvious eco-
CONCLUSIONS nomic returns to the largely least developed carob-produc-
Laboratory-made carob syrups contained more proteins ing countries. However, the age-old practice of heat concen-
than date and maple syrups and more minerals (Ca, Mg, Na, tration in open vats leads to excessive browning and the
K and P) than maple syrup and, in general, higher or at least concomitant formation of high levels of the potentially mu-
similar mass fractions of the indicated minerals than date syr- tagenic and carcinogenic hydroxymethylfurfural and, there-
up. The carob syrups had sugar compositions intermediate fore, milder heating regimens are warranted in the produc-
to those of the invert-sugar-like composition of date syrup tion of carob syrups.
and the overwhelmingly sucrose-containing maple syrup.
Furthermore, the vast majority of carob syrups contained
ACKNOWLEDGEMENTS
more total phenols than maple syrup and higher, or at least
The technical assistance of the personnel at the Kamal
comparable, levels of total phenols than date syrup. Similar
Shair Central Research Science Laboratory, at the American
patterns to those of total phenols were found for the antiox-
University of Beirut (AUB), in the analysis of minerals and Mr.
idant capacity of the syrups by in vitro antioxidant capacity
Hasan El Nisr from the Office of Communications, at AUB, for
assays. Gallic acid was the major phenolic acid in the carob,
his expertise in capturing the stages of carob syrup produc-
date and maple syrups, and, in general, the carob syrups con-
tion is gratefully acknowledged.
tained more phenolic acids and flavonoids than the other syr-
ups. The carob syrups browned more and generated more
hydroxymethylfurfural than maple syrup during the heat FUNDING
concentration step and, in general, were less brown and con- The financial support provided by the University Re-
tained lower mass fractions of hydroxymethylfurfural than search Board of the American University of Beirut (Grant no.
date syrup. These findings indicate that carob syrup exhibits 24620-103601) is gratefully acknowledged.

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I. TOUFEILI et al.: Chemical and Antioxidant Profiles of Carob Syrup

CONFLICT OF INTEREST natural sweeteners and evaluation of their metabolic re-

The authors declare no conflict of interest. sponses in healthy rats. J Funct Foods. 2014;11:460–471.
https://doi.org/10.1016/j.jff.2014.10.001
 9. Singh AS, Jones AMP, Saxena K. Variation and correlation
SUPPLEMENTARY MATERIALS
of properties in different grades of maple syrup. Plant
Supplementary material is available at www.ftb.com.hr.
Foods Hum Nutr. 2014;69(1):50–6.
https://doi.org/10.1007/s11130-013-0401-x
AUTHORS’ CONTRIBUTION 10. Fattouch S, Dhaouadi K, Belkhir M. Date syrup. In: Shahidi
I. Toufeili conceptualized the research, supervised the F, Alasalvar C, editors. Handbook of functional beverages
chemical analyses, secured funding and drafted the manu- and human health. Boca Raton, FL, USA: CRC Press; 2016.
script. M. Itani and M. Zeidan performed the chemical analy- pp. 217–29.
ses and processed the collected data. O. Al Yamani developed https://doi.org/10.1201/b19490
and carried out the HPLC analyses. S. Kharroubi worked out
11. Nasar-Abbas SM, Huma ZE, Vu TH, Khan MK, Esbenshade
the statistical design and supervised the statistical analyses.
H, Jayasena V. Carob Kibble: A bioactive-rich food ingre-
All authors have read and approved the final manuscript. I.
dient. Compr Rev Food Sci Food Saf. 2016;15:63–72.
Toufeili approved the final version of the manuscript.
https://doi.org/10.1111/1541-4337.12177
12. Rodríguez-Solana R, Romano A, Moreno-Rojas JM. Carob
ORCID ID
pulp: A nutritional and functional by-product worldwide
I. Toufeili https://orcid.org/0000-0002-5722-3999 spread in the formulation of different food products and
M. Itani https://orcid.org/0000-0002-5494-9161 beverages. A review. Process. 2021; 9(7):1146.
M. Zeidan https://orcid.org/0000-0003-0299-4400 https://doi.org/10.3390/pr9071146
O. Al Yamani https://orcid.org/0000-0002-9344-2427
13. Dhaouadi K, Belkhir M, Akinocho I, Raboudi F, Pamies D,
S. Kharroubi https://orcid.org/0000-0002-2355-2719
Barrajón E, et al. Sucrose supplementation during tradi-
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