Accepted Manuscript

Title: Chemical characterization of fructooligosaccharides,

inulin and structurally diverse polysaccharides from
chamomile tea

Authors: Pedro Felipe P. Chaves, Marcello Iacomini,

Lucimara M.C. Cordeiro

PII: S0144-8617(19)30322-4
DOI: https://doi.org/10.1016/j.carbpol.2019.03.050
Reference: CARP 14719

To appear in:

Received date: 7 February 2019

Revised date: 11 March 2019
Accepted date: 14 March 2019

Please cite this article as: Chaves PFP, Iacomini M, Cordeiro LMC,
Chemical characterization of fructooligosaccharides, inulin and structurally
diverse polysaccharides from chamomile tea, Carbohydrate Polymers (2019),
https://doi.org/10.1016/j.carbpol.2019.03.050

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Chemical characterization of fructooligosaccharides, inulin and

structurally diverse polysaccharides from chamomile tea

Pedro Felipe P. Chaves, Marcello Iacomini, Lucimara M.C. Cordeiro*

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Department of Biochemistry and Molecular Biology, Federal University of Paraná, CP

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19.046, CEP 81.531-980 Curitiba- PR, Brazil

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∗ Corresponding author. Tel.: +55 41 33611655; fax: +55 41 32662042.
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E-mail address: lucimaramcc@ufpr.br (L.M.C. Cordeiro).
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HIGHLIGHTS
 Polysaccharides from the chamomile tea were purified and characterized.
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 Inulin, HM homogalacturonan and a type II arabinogalactan were obtained.

 Fructooligosaccharides ranging from GF2 (m/z 543) to GF10 (m/z 1839) were
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also present.
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 Chamomile is a source of structurally diverse dietary fibers.

 Chamomile tea polysaccharides could have potential prebiotic function.

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ABSTRACT

Chamomile is one of most known species of medicinal plants. It has valuable

pharmacological properties that produce positive effects in many therapeutical uses.

Some of these properties are attributed to the presence of secondary metabolites but is

already known that primary metabolites can also produce positive effects. In this study

we elucidate the fine chemical structure of polysaccharides present in the infusion of

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chamomile flower chapters. After ethanolic precipitation, polysaccharides were

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obtained from the tea (fraction MRW, 3.2% yield), purified and characterized as an

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inulin type fructan, a highly methyl esterified and acetylated homogalacturonan (DE =

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87% and DA = 19%), and a type II arabinogalactan. From ethanolic supernatant (20.2%

yield), fructooligosaccharides (FOS) ranging from GF2 (m/z 543) to GF10 (m/z 1839)
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were detected. Inulin and FOS are well-established prebiotics, as well as the pectic
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polysaccharides. Thus, chamomile could be a source of structurally diverse dietary
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fibers with potential prebiotic, gastrointestinal and immunological functions.

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Keywords: chamomile tea; inulin; fructooligosaccharides; homogalacturonan,

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arabinogalactan.
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1.Introduction

Medicinal plants have a fundamental role in the world health, they can be used

as sources of direct therapeutic agents, can serve as a raw material for the elaboration of

semi-synthetic pharmaceuticals or the discovery of new compounds (Akerele, 1993).

Hence, every year more species have their chemical components described, their

therapeutic effectiveness are proven and also the discovery of new therapeutic uses

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occurs (Halberstein, 2005).

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Numberless species are explored for their pharmacological effects, among them

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are the chamomile. Chamomilla recutita [L.] Rauschert, commonly called German

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chamomile, is one of most known medicinal species and is included in the

pharmacopoeia of almost all countries (Franke & Schilcher, 2005). It is consumed in

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infusion or decoction form from its floral chapters, to obtain the positive effects as
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improver of digestion, to facilitate the elimination of gases, to stimulate the appetite, to
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relief anxiety, to treat colic, wounds or diseases of the skin, as healing agent and mainly
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as an anti-inflammatory medicine (Lorenzi & Matos, 2008; Sousa, Matos, Matos,

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Machado, & Craveiro, 1991). Moreover, the chamomile oil is extensively used in

perfumery, cosmetics, aromatherapy and in pharmaceutical and food industries. Thus,

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there is a great demand for chamomile in the market and it is the fifth top selling herb in

the world (Singh, Khanam, Misra, & Srivastavaet, 2011).

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The pharmacological properties exhibit by medicinal plants are usually

attributed to the presence of specific secondary metabolites, however it is already

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known that some primary metabolites, such as polysaccharides, can work together to

produce these properties and also can exhibit strong biological effects per se

(Halberstein, 2005; Liu, Willför, & Xu, 2015).

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Polysaccharides can also have prebiotic effect (Roberfroid, 2007a). Their

capacity of escaping the digestion in the upper gastrointestinal tract and become

available for fermentation by microbiota is already known and can be linked to their

structural characteristics, such as monosaccharide composition, glycosidic bond

configuration, amount and size of branches and molar mass (Cantu-Jungles, Cipriani,

Iacomini, Hamaker, & Cordeiro, 2017; Roberfroid, 2007a, 2007b). Thus, in the present

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study we described the purification process of polysaccharides obtained from

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chamomile infusion, its structural characterization and with the results we suggested a

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new therapeutical use to the species, as a source of prebiotic polysaccharides.

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2. Material and Methods
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2.1. Plant material
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Dried floral C. recutita chapters were kindly provided by Chamel® Produtos

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Naturais Industry. The plant material was stored in a sealed plastic container at room
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temperature until use. In addition, a voucher specimen of industry’s crop was collected

(Campo Largo - PR, Brazil, 25º24.58’’S 49º27.64’’W, in 2013 September) to confirm

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the botanical identity and deposited in the Museu Botânico Municipal de Curitiba, under

registration number 382674.

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2.2. Extraction of polysaccharides

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The floral chapters were reserved in a beaker and boiling distilled water was

added (40 g/L), the beaker was closed and let rest for about 30 minutes. The extract

(tea) was filtered, concentrated under reduced pressure and the polysaccharides

precipitated with 95% ethanol (3 vol.). The polysaccharides were recovered by

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filtration, dialyzed in semipermeable membrane (Cellulose Spectrumlabs 6-8 kDa cut-

off) and freeze-dried (MRW fraction) (Fig. 1). These procedures were repeated several

times to enable the extraction of 628 g of floral chapters.

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Figure 1. Scheme of extraction and purification of polysaccharides from infusion of

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Chamomilla recutita floral chapters.

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MRW was further fractionated by ultrafiltration on 100 kDa cutoff membrane

(Fig. 1), giving MRW-100R (retained on the membrane) and MRW-100E (eluted). This
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latter was ultrafiltrated on 30 kDa membrane. The retained fraction (MRW-30R) was
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treated with Fehling solution (Jones & Stoodley, 1965), and the resulting insoluble Cu2+
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complex isolated by centrifugation. Both (Fehling supernatant and precipitated

fractions, SF and PF, respectively) were neutralized with acetic acid, dialyzed, and

deionized with H+ form cation-exchange resin. SF was then treated with endo-inulinase

enzyme (316 U/mg, Megazyme) in acetic acid/sodium acetate buffer (pH 4.6) for 16
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hours at 45 ºC and then dialyzed (Cellulose Spectrumlabs 6-8 kDa cut-off), giving SF-

EN fraction. Finally, it was submitted to anion exchange chromatography on DEAE

Sepharose Fast Flow (GE Healthcare) and eluted with water, to give fraction SF-EN-

AG. All the fractionation steps are summarized in Fig. 1. Yields of polysaccharide

fractions were expressed as percent based on the weight of dried floral chapters that

were submitted to extraction (628 g).

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2.3. Determination of monosaccharide composition

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All fractions (except MRW-30E) were hydrolyzed in 500 μL 2 M TFA at 100 °C

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for 8 hours. MRW-30E was hydrolyzed with 500 μL 0.2 M TFA at 80°C for 30 min.

The TFA was evaporated and the samples were converted to alditol acetates by NaBH4
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reduction at 100 ºC for 10 minutes followed by acetylation with Ac2O-pyridine (1:1,
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v/v, 1 mL) at 100 ºC for 30 minutes. The resulting alditol acetates were then extracted
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with CHCl3 and analyzed by GC-MS using a Varian 3800 gas chromatograph coupled
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to a Varian Ion-Trap 2000R mass spectrometer (Varian, Palo Alto, CA). The column
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was DB-225MS (30 m 0.25 mm i.d.; Agilent Santa Clara, CA) programmed from 50 to

220 °C at 40 °C/min, with helium as carrier gas, at a flow rate of 1 mL/min. The inlet
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temperature was 250 °C, and the MS transfer line was set at 250 °C. MS acquisition

parameters included scanning from m/z 50 to 550 in electron ionization mode (EI) at 70
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eV. Components were identified by their retention times and EI spectra. Fructose upon

reduction and acetylation gives glucitol and mannitol acetates on GC-MS analysis. The
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amounts of both derivatives have been summed up to give the amount of fructose

present in the sample.

Uronic acid contents were determined using the modified m-hydroxybiphenyl

method (Filisetti-Cozzi & Carpita, 1991).

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2.4. Determination of homogeneity and relative molecular weight

The homogeneity and relative molecular weight (Mw) of water-soluble

polysaccharides were evaluated by high performance steric exclusion chromatography

(HPSEC), with a Waters 2410 differential refractometer as equipment for detection. A

series of four columns, with exclusion sizes of 7 x 106 Da (Ultrahydrogel 2000, Waters),

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4 x 105 Da (Ultrahydrogel 500, Waters), 8 x 104 Da (Ultrahydrogel 250, Waters) and 5 x

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103 Da (Ultrahydrogel 120, Waters) was used. The eluent was 0.1 M aq. NaNO2

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containing 200 ppm aq. NaN3 at 0.6 mL/min. The sample, previously filtered through a

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membrane (0.22 µm, Millipore), was injected (250 µl loop) at a concentration of 1

mg/mL. To obtain the relative Mw, standard dextrans (487 kDa, 266 kDa, 124 kDa, 72.2
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kDa, 40.2 kDa, 17.2 kDa and 9.4 kDa, from Sigma) were employed to obtain the
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calibration curve. The relative Mw of the sample was calculated according to the
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calibration curve.
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2.5. Methylation analysis

Fraction SF-EN-AG was carboxyl reduced by the carbodiimide method, using

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NaBH4 as the reducing agent, giving products with the –COOH groups of its uronic acid

residues reduced to –CH2OH (Taylor & Conrad, 1972). The carboxyl reduced sample
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was O-methylated according to Ciucanu and Kerek (1984) method, using powdered

NaOH in DMSO-MeI. The per-O-methylated polysaccharide was then submitted to

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methanolysis in 3% HCl–MeOH (80 °C, 2 h) followed by hydrolysis with H2SO4

(0.5M, 12 h) and neutralization with BaCO3. The material was then submitted to

reduction and acetylation as described above for sugar composition, except that the

reduction was performed using NaBD4. The products (partially O-methylated alditol
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acetates) were examined by capillary GC-MS. A capillary column (30 m x 0.25 mm

i.d.) of DB-225, held at 50 ºC during injection for 1 min and then programmed at 40

ºC/min to 210 ºC and held at this temperature for 31 min, was used for separation. The

partially O-methylated alditol acetates were identified by their typical electron impact

breakdown profiles and retention times (Sassaki, Gorin, Souza, Czelusniak, & Iacomini,

2005).

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2.6. Nuclear magnetic resonance spectroscopy

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The 1H, 13C and heteronuclear single quantum coherence (HSQC-DEPT 135)

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spectra were obtained from samples dissolved in D2O, at 70 °C using a 400 MHz

Bruker model DRX Avance III spectrometer, operating at 9.5 T, observing 1H at 400.13
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MHz and 13C at 100.61 MHz, equipped with a 5-mm multinuclear inverse detection
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probe with z-gradient. The chemical shifts are expressed in ppm relative to CH3 signal
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from internal reference acetone (δ 30.2/2.22). All pulse programs were supplied by
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Bruker.
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2.7. Electrospray ionization mass spectroscopy analysis

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A syringe pump was used at a flow rate of 5 μL/min to infuse fraction MRW-ET

(at 200 µg/mL) directly into the mass spectrometer. The positive high-resolution mass
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spectroscopy analysis was carried out with electrospray ionization (ESI) at atmospheric

pressure ionization (API) in an LTQ-OrbiTrap-XL (Thermo-Scientific), using N2 for

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sample desolvation with sheath gas at a flow rate of 8 UA and auxiliary gas at 2 UA

with a source temperature of 300°C. The ionization was performed following the

operational parameters: electrospray voltage at 4 kV, capillary voltage 25 V, tube lens

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offset 125 V. The spectra were processed and analysed with Thermo Xcalibur 1.0.0.42

software.

3.Results and discussion

The process of extraction by infusion of C. recutita floral chapters produced a

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crude polysaccharide fraction named MRW with 3.2% yield from the dry weight and an

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ethanolic supernatant (MRW-ET, 20% yield). The sugar composition, which showed

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uronic acids, arabinose, galactose, xylose, rhamnose and fructose (Table 1) together

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with HSQC correlation map analysis of MRW (Fig. 2) allowed a preliminary

identification of two main polysaccharide types present in chamomile tea: (1) a methyl
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esterified homogalacturonan (HG) could be detected due to the signals at δ 100.0/4.97
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(C1-H1 from methyl esterified GalpA), δ 99.3/5.18 (C1-H1 from GalpA), δ 68.0/3.75
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(C2), δ 68.3/3.98 (C3), δ 78.6/4.46 (C4), δ 70.5/5.05 (C5 from methyl esterified GalpA)
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and δ 52.8/3.82 (-COOCH3); and (2) a fructan of inulin-type, due to the signals at δ
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61.0/3.73 (C1-H1), δ 103.2 (C2, visible only in the 13C spectrum, data not shown), δ

77.2/4.23 (C3-H3), δ 74.6/4.09 (C4-H4), δ 81.1/3.86 (C5-H5) and δ 62.0/3.76 and 3.83
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(C6-H6) (Corrêa-Ferreira, Noleto, & Oliveira Petkowicz, 2014; A. J. B. de Oliveira et

al., 2011; Perrone et al., 2002; Sergey V. Popov et al., 2011; Vriesmann & de Oliveira
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Petkowicz, 2009). Small amounts of an arabinogalactan may also be present by the

observed anomeric signals of β-D-Galp units at δ 102.9/4.47 and that of α-L-Araf at δ

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107.6/5.07 and δ 109.0/5.25 (Nascimento, Iacomini, & Cordeiro, 2017; A. F. de

Oliveira, Nascimento, Iacomini, Cordeiro, & Cipriani, 2017).

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Figure 2. HSQC correlation map of MRW fraction in D2O at 70 °C, the chemical shifts

are expressed as δ ppm. Ara = arabinose, GalA = galacturonic acid, GalA’= methyl
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esterified galacturonic acid, Fru = fructose.

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These two main polysaccharide types were also observed in homogeneity analysis,

where a heterogeneous profile with two evident peaks (I and II) (Fig. 3) were present.
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To isolate them, the fraction was submitted to ultrafiltration using a 100 kDa cutoff

membrane. The process was highly efficient, once peak II was concentrated in the
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eluted fraction (MRW-100E, 1.4% yield), while peak I remained retained on the

membrane (MRW-100R, 1.0% yield). This latter contained the pectic

homogalacturonan. It had mainly uronic acid (Table 1) on sugar analysis, identified as

galacturonic acid by GC-MS of carboxyl-reduced sample, and a relative Mw of 500 kDa.

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13
C NMR spectrum (Fig. 4A) showed typical signals of the methyl esterified HG (as

cited above). The degree of methyl esterification was determined by 1H NMR following

the method of Grasdalen, Bakøi e Larsen (1988) giving a value of 87%, characterizing

the chamomile pectin as a HM pectin (Silva & Rao, 2006). Due to the presence of acetyl

signals at δ 20.3 in the 13C NMR spectrum, the degree of acetylation was also

determined by 1H NMR following the method of An et al. (2011) and

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spectrophotometrically by Hestrin (1949) methodology, giving a value of 19%.

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Figure 3. HPSEC elution profile of fractions MRW, MRW-100E and MRW-100R.

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Refractive index detector. Elution volume of dextran standards of molecular weight 487

kDa, 266 kDa, 124 kDa, 72.2 kDa, 40.2 kDa, 17.2 kDa and 9.4 kDa (left to right) were
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employed to construct the calibration curve.

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Figure 4. 13C NMR spectra of fractions MRW-100R (A), MRW-100E (B) and SF-EN
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(C) in D2O at 70 °C, the chemical shifts are expressed as δ ppm.

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Fraction MRW-100E containing the peak II of MRW (with relative Mw < 9.4

kDa) also showed a third small peak in HPSEC analysis (with relative Mw of 60 kDa)

(Fig. 3) and thus was submitted to a new ultrafiltration procedure using a 30 kDa cutoff

membrane. Peak II was eluted in the membrane (MRW-30E fraction) and had fructose
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on sugar analysis as the major constituent (Table 1). 13C NMR analysis (Fig. 4B)

indicated the presence of the inulin-type fructan, with six typical signals of →1)-β-D-

Fruf-(2→ at δ 61.2 (C1), δ 103.2 (C2), δ 77.5 (C3), δ 74.8 (C4), δ 81.3 (C5) and δ 62.3

(C6) (Corrêa-Ferreira et al., 2014; A. J. B. de Oliveira et al., 2011). Looking for the

presence of fructooligosaccharides (FOS) in chamomile tea, we analyzed fraction

MRW-ET, which was obtained in high yield (20%), using the LTQ Orbitrap-XL Hybrid

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Ion Trap-Orbitrap Mass Spectrometer. The MS spectra (Fig. 5) showed besides sucrose,

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FOS ranging from GF2 (m/z 543) to GF10 (m/z 1839).

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Figure 5. MS spectra (+ve mode) of MRW-ET fraction obtained in LTQ Orbitrap-XL

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Hybrid Ion Trap-Orbitrap Mass Spectrometer.

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Thus, the results showed that chamomile tea contains as main polysaccharides a

highly methyl esterified and acetylated homogalacturonan and inulin, besides high

amounts of fructooligosaccharides. A previous study about C. recutita polysaccharides

pointed out the existence of a polysaccharide containing (1→4)-linked α-D-GalpA

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residues (Yakovlev & Gorin, 1977), but the structural characterization of the polymer

has not been performed by the authors. Later, Füller and Franz (1993) observed the

presence of a fructan of the inulin type in their C. recutita extracts, but the presence of

FOS in chamomile tea has not been reported in the literature yet. Fructans are

commonly found in species from the Asteraceae family, to which C. recutita belongs.

These can be found as reserve polymers in the tuberous roots of Jerusalem artichoke

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(Helianthus tuberosus) (Saengthongpinit & Sajjaanantakul, 2005), chicory (Cichorium

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intybus) (Toneli, Park, Ramalho, Murr, & Fabbro, 2008) and yacon (Smallanthus

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sonchifolius) (Paredes et al., 2018). In the aerial parts, fructans have already been found

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in artemisia (Artemisia vulgaris) (Corrêa-Ferreira et al., 2014), stevia (Stevia

rebaudiana) (de Oliveira et al., 2011) and another Matricaria species (M. maritima
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(Cérantola et al., 2004). They were also extracted from the monocotyledon agave plant
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(Agave tequilana var. azul) (Praznik, Löppert, Cruz Rubio, Zangger, & Huber, 2013).
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It is well stablished in the literature that inulin is a versatile substance with

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numerous health benefits. Inulin and FOS are the most studied and well-established
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prebiotics. They escape digestion in the upper gastrointestinal tract and reach the large

intestine virtually intact, where they modulate the composition and activities of the gut
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microbiota (Roberfroid, 2007a). Moreover, it has been demonstrated that pectic

polymers from different sources can also be prebiotics, being extensively fermented in
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the colon and are able to modulated the gut microbiota (Cantu-Jungles et al., 2017;

Gulfi, Arrigoni, & Amadò, 2005; Jonathan et al., 2012; Licht et al., 2010; Min et al.,
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2015; Titgemeyer, Bourquin, Fahey, & Garleb, 1991). It is worth noting that inulin,

FOS and pectins can also specifically affect several other gastrointestinal functions (for

example, mucosal functions, endocrine activities and mineral absorption) as well as

systemic functions (especially glucose and lipid homeostasis and immune functions)
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(Lunn & Buttriss, 2007; S V Popov & Ovodov, 2013; Roberfroid, 2007a; Vogt et al.,

2015).

To a comprehensive identification of chamomile polysaccharides, the low-yield

fraction MRW-30R which corresponded to the peak III (Fig. 3) was also chemically

characterized. It had a very complex monosaccharide composition, composed of

rhamnose, arabinose, xylose, fructose, galactose and uronic acid (Table 1). Galacturonic

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acid and fructose came from HG and inulin, that were still present in this fraction

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(observed in its 13C NMR spectrum, data not shown). To further purification and

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characterization of other polysaccharides, MRW-30R was treated with Fehling reagent

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once homogalacturonans interact with copper and precipitate. Thus, due to alkaline pH

of Fehling reagent, deesterified and deacetylated HG remained in PF fraction, as could

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be observed in its 13C NMR spectrum (Suppl. Fig 1). Fraction SF was also treated with
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endo-inulinase, due to the presence of some amounts of contaminating inulin. On sugar
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analysis, fraction SF-EN presented rhamnose, arabinose, galactose, xylose and uronic
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acids (Table 1). Its 13C NMR spectrum (Fig. 4C) showed signals at δ 101.1 and δ 101.7
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assigned to anomeric β-D-Xylp units, and at δ 97.6 (C1) and 59.4 (-OCH3) assign to 4-

O-Me-α-D-GlcpA units, probably from an acid xylan (Dinand & Vignon, 2001; Vignon
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& Gey, 1998), and signals at δ 103.4 (anomeric carbon of β-D-Galp) and at δ 107.6 and

δ 109.0 (anomeric carbons of α-L-Araf units), probably from an arabinogalactan

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(Nascimento et al., 2017; A. F. de Oliveira et al., 2017). Finally, fraction SF-EN was

further purified by anion exchange chromatography in DEAE Sepharose Fast Flow. The
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column was eluted with water, giving a fraction (SF-EN-AG) composed mainly of

galactose and arabinose (Table 1). Methylation analysis of carboxyl reduced sample

(Table 2) confirmed the presence of an arabinogalactan. The main methylated derivative

was 2,4-Me2-Gal-ol acetate, from 3,6-di-O-substituted Galp units. Other Gal derivatives
 16

were 2,3,4,6-Me4-Gal-ol, 2,3,4-Me3-Gal-ol, 2,4,6-Me3-Gal-ol and 4-Me-Gal-ol acetates,

from terminal, 6-O-, 3-O- and 2,3,6-tri-O-substituted Galp units, respectively.

Arabinose was present as terminal, 5-O- and 3,5-di-O-substituted Araf units. Terminal

Glcp units were also observed, from GlcpA units. Its HSQC-DEPT correlation map

(Fig. 6) showed anomeric cross peaks at  109.0/5.24 and  107.3/5.07 from terminal

and →5)-α-L-Araf-(1→ units, at  103.8/4.69,  103.2/4.46 and  103.0/4.51 from

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terminal, →3)-β-D-Galp-(1→/→3,6)-β-D-Galp-(1→ and →6)-β-D-Galp-(1→,

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respectively. Inverted DEPT signals were at  69.2/3.92-4.04 from 6-O-linked β-D-Galp

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units and at  66.6/3.80-3.87 from 5-O-linked α-L-Araf units. Other inverted signals

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were at  61.2/3.80,  61.1/3.73 and  60.9/3.77 from unsubstituted C-6/H-6 or C-5/H-5

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from α-L-Araf-(1→, β-D-Galp-(1→ and →3)-β-D-Galp-(1→ units. The assignments are
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in agreement with published literature data and methylation analysis described above
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(Brecker et al., 2005; Capek, Matulová, Navarini, & Suggi-Liverani, 2010; Dong &
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Fang, 2001; Goellner, Utermoehlen, Kramer, & Classen, 2011; Liang, Hu, He, & Pan,

2014; H. Wang, Shi, Bao, Li, & Wang, 2015; P. Wang et al., 2015) and shows the
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presence of a type II arabinogalactan in SF-EN-AG fraction. In their preliminary

characterization of C. recutita polysaccharides, Füller and Franz (1993) also suggested

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the presence of a rhamnogalacturonan with type II arabinogalactan and a

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glucuronoxylan in the aqueous chamomile extracts. However, the fine chemical

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structure of these polysaccharides had not been determined.

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Figure 6. HSQC-DEPT correlation map of SF-EN-AG fraction in D2O at 50 °C, the

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chemical shifts are expressed as δ ppm. Inverted signals in DEPT experiment are shown

in blue color.
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Matricaria chamomilla belongs to a major group of cultivated medicinal plants,

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often referred to as the “star among medicinal species”. More than 120 chemical

constituents have been identified in chamomile flower as secondary metabolites, which

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gives to chamomile its multitherapeutic, cosmetic, and nutritional values, that have been

established through years of traditional and scientific use and research (Singh, Khanam,

Misra, & Srivastava, 2011). The presence of inulin, FOS, highly methyl esterified

homogalacturonan, type II arabinogalactan and acid xylan in chamomile tea shows that
 18

not only can the secondary metabolites be the responsible molecules by the health

benefits of chamomile consumption and adds to chamomile a new property, as a source

of structurally diverse dietary fibers with potential prebiotic, gastrointestinal and

immunological functions.

Acknowledgements

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This research was supported by CAPES (Process 1264763), Fundação Araucária

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and by Universal Project (Process 404717/2016-0) provided by CNPq foundation

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(Brazil). The authors are grateful to Chamel® Produtos Naturais Industry who kindly

provided the dried floral C. recutita chapters, to the NMR Center of UFPR for recording
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the NMR spectra and to Dr. Lauro M. de Souza for the mass spectroscopy analysis.
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Table 1

Monosaccharide composition of fractions obtained from chamomile (C. recutita) tea.

Neutral sugarsa Uronic

Fraction
Rha Ara Xyl Gal Fruc acidb

MRW 3.0 24.6 8.6 12.0 12.6 39.2

T
MRW-100R 1.1 4.0 1.9 2.2 - 91.0

IP
MRW-30E - 4.7 - - 95.3d nde

R
MRW-30R 2.6 18.5 6.9 11.6 16.3 44.1

SC
SF-EN 3.0 40.6 18.4 24.0 tr 14.0

SF-EN-AG - 37.4 - 58.0 - 4.6

a U
% of peak area relative to total peak area, determined by GC-MS.
N
b
Determined using the m-hydroxybiphenyl method (Filisetti-Cozzi & Carpita, 1991).
A
c
The amounts of glucitol and mannitol acetates on GC-MS analysis have been summed up to give the
M

amount of fructose present in the sample.

d
Hydrolysis with 0.2 M TFA at 80°C followed by GC-MS analysis.
ED

e
Not determined.
E PT
CC
A
 28

Table 2

Linkage types based on analysis of partially O-methyl alditol acetates obtained from

methylated type II arabinogalactan (fraction SF-EN-AG) from chamomile (C. recutita)

tea.

Partially O- SF-EN-AGa Linkage typeb

methylalditol acetate

T
IP
2,3,5-Me3-Araf c 11.5 Araf-(1→

R
2,3,4,6-Me4-Glcp 7.2 Glcp-(1→

SC
2,3,4,6-Me4-Galp 14.9 Galp-(1→

2,3-Me2-Araf 8.8
U →5)-Araf-(1→
N
2-Me-Araf 1.1 →3,5)-Araf-(1→
A

2,4,6-Me3-Galp 2.3 →3)-Galp-(1→

M

2,3,4-Me3-Galp 4.1 →6)-Galp-(1→

ED

2,4-Me2-Galp 45.2 →3,6)-Galp-(1→

PT

4-Me-Galp 4.9 →2,3,6)-Galp-(1→

a
Fraction was carboxyl reduced by Taylor and Conrad (1972) method. % of peak area of O-methyl alditol
E

acetates relative to total area, determined by GC-MS.

CC

b
Based on derived O-methyl alditol acetates.
c
2,3,5-Me3-Ara = 2,3,5-tri-O-Methylarabinitolacetate, etc.
A