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Metabolomic pro
file of Salvia viridis L. root extracts using HPLC–MS/MS
technique and their pharmacological properties: A comparative study
Gokhan Zengin
a,⁎, Fawzi Mahomoodally
b, Carene Picot-Allain
b, Alina Diuzheva
c, József Jek
ő
d,
Zoltán Cziáky
d, Aleksandra Cvetanovi
ć
e, Abdurrahman Aktumsek
a, Zoran Zekovi
ć
e,
Kannan R.R. Rengasamy
faDepartment of Biology, Science Faculty, Selcuk University, Campus, Konya, Turkey bDepartment of Health Sciences, Faculty of Science, University of Mauritius, Réduit, Mauritius cDepartment of Analytical Chemistry, Pavol JozefŠafárik University in Košice, Košice, Slovakia
dAgricultural and Molecular Research and Service Institute, University of Nyíregyháza, Nyíregyháza, Hungary eFaculty of Technology, Bulevar Cara Lazara 1, 21000, Novi Sad, Serbia
fREEF Environmental Consultancy, #2 Kamaraj Street, S.P. Nagar, Puducherry, 605 001, India
A R T I C L E I N F O Keywords: Salvia viridis Diabetes Alzheimer’s disease Hyperpigmentation Extraction Phyto-pharmaceuticals A B S T R A C T
Several Salvia species have received due scientific attention regarding their therapeutic virtues, yet little is known about the pharmacological potential of Salvia viridis L. roots. This study, therefore, attempts to explore the phytochemical composition, enzyme inhibitory potential, and antioxidant activities of S. viridis ethanolic root extracts obtained by different extraction methods, namely microwave-assisted extraction, maceration,
su-percriticalfluid extraction, Soxhlet extraction, and ultrasonic assisted extraction. The extract produced by
ul-trasonic assisted extraction possessed the highest phenolic and flavonoid contents (111.41 mg gallic acid
equivalent/g extract and 23.46 mg rutin equivalent/g extract). S. viridis ethanolic root extract obtained by ul-trasonic assisted extraction showed highest radical scavenging (240.00 and 302.85 mg Trolox equivalent TE/g
for DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2′-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid)
assays, respectively) and reducing (970.74, 704.27 mg TE/g, and 2.84 mmol TE/g for CUPRAC (cupric reducing antioxidant capacity), FRAP (ferric reducing antioxidant power), and phosphomolybdenum assays, respectively)
activities. Chemical profiles of these extracts were investigated by HPLC–MS/MS, and the profiles (23
compo-nents) of the supercriticalfluid extract was different from other extraction techniques. The study reports for the
first time, the inhibitory action of ethanolic root extract of S. viridis on key enzymes related to Alzheimer’s
disease (acetylcholinesterase, butyrylcholinesterase), diabetes (α-amylase, α-glucosidase), and skin
hy-perpigmentation disorders (tyrosinase). Data generated from this study appraises the multiple biological
ac-tivities of plants belonging to the Salvia genus. Scientific evidence gathered in this study support further
in-vestigations which might lead to the development of new pharmaceutical entities for the management of diabetes, Alzheimer’s disease, and skin hyperpigmentation conditions.
1. Introduction
It is generally agreed that medicinal plants represent promising
al-ternatives for the management of numerous human ailments. Multiple
lines of evidence attest of the ethnopharmacological use of plants by
several folk populations since ancient times (
Zengin et al., 2019
).
In-deed, naturally occurring plant compounds underpin drug
develop-ment. It is noteworthy highlighting that nearly one-quarter of new
molecular entities were derived from naturally occurring compounds
(
Patridge et al., 2016
;
Mollica et al., 2017
). Additionally, the rising
prevalence of besetting diseases, along with the ever-increasing demand
for more effective and safer drugs, provide further impetus for research.
The type of extraction method is one of the most critical steps in the
phytochemical studies, and thus several techniques have been
sidered to extract phytochemicals from plants. In recent years,
con-ventional methods (maceration, soxhlet, etc.) have been replaced by
advanced techniques including ultra-sonication, microwave, and
su-percritical methods, which have several advantages such as
eco-friendly, reduce use of solvents and extraction times. In this respect,
many researchers have been focusing on the best extraction techniques
https://doi.org/10.1016/j.indcrop.2019.01.060
Received 13 December 2018; Received in revised form 26 January 2019; Accepted 28 January 2019
⁎Corresponding author.
E-mail address:gokhanzengin@selcuk.edu.tr(G. Zengin).
Industrial Crops & Products 131 (2019) 266–280
Available online 05 February 2019
0926-6690/ © 2019 Elsevier B.V. All rights reserved.
with high efficiency of solvents (
Ameer et al., 2017
;
Belwal et al.,
2018
).
The Salvia genus is the most prominent genus of the Lamiaceae
fa-mily and it contains approximately 900 species. Species of this genus
are widely distributed in temperate, subtropical, and tropical regions
(
Shari
fi-Rad et al., 2018
). The most common applications of Salvia
species include the food, cosmetics, and pharmaceutical industries.
Several genera of this species have been used in traditional medicine,
for instance, S. cavaleriei has been used to treat dysentery, boils,
in-juries, haemoptysis; S. desoleria has been used to treat central nervous
system, menstrual, and digestive problems; S. bucharica has been used
to manage liver disorders (
Hao et al., 2015
). Salvia species have been
reported to possess multiple bio-pharmaceutical activities, namely,
antioxidant,
antimicrobial,
antidiabetic,
neuroprotective,
anti-in-flammatory, and cytotoxic properties (
Zengin et al., 2018a
).
Salvia viridis L. (synonym Salvia horminum L.), commonly known as
‘Red topped sage’, naturally occurs in the Mediterranean region. It is a
perennial, annual or biennial herb, having the erect stem of 50 cm and
4
–8 axillary flowers (
Rungsimakan and Rowan, 2014
). S. viridis has
been used in traditional medicine as gargle against sore gum
(
Grzegorczyk-Karolak and Kiss, 2018
). In Turkey, an infusion of the
shoot,
flowers, and leaves of S. viridis have been used against a sore
throat, throat in
flammation, antitussive, ulcer, intestinal spasm, and
gynaecological complications (
Sharifi-Rad et al., 2018
). According to
our literature search, no study has reported the inhibitory activity of S.
viridis roots on key enzymes related to skin hyperpigmentation
pro-blems, Alzheimer’s disease, and diabetes. Besides, this study evaluates
the possible e
ffect of different extraction techniques on the bioactivity
of S. viridis roots. It is anticipated that data generated by this study will
provide new horizons into the possible biological activity of S. viridis a
poorly studied medicinal plant from the Salvia genus.
2. Materials and methods
2.1. Collection of plant material
Salvia viridis roots were collected in Adana-Turkey, in June 2017.
The taxonomic identi
fication was performed by the botanist Dr. Murad
Aydin Sanda (Muş Alpaslan University, Department of Molecular
Biology, Muş, Turkey) and one voucher specimen was deposited at the
herbarium of Selçuk University, Konya, Turkey. The roots were dried at
the room temperature until a constant weight was recorded. The
sam-ples were then ground using a laboratory mill.
2.2. Extraction techniques
2.2.1. Microwave-assisted extraction (MAE)
Five grams of root sample was extracted with 100 mL of 96% (w/w)
ethanol (1:20 ratio). The extractions were performed in an open system
at 600 W during 30 min.
2.2.2. Maceration (MAC)
To produce macerated extracts, the root samples (5 g) were
ma-cerated with 100 mL of ethanol at room temperature for 24 h.
2.2.3. Supercritical
fluid extraction (SFE)
SFE extraction of the root samples was performed under the
fol-lowing conditions: 50 °C, 350 bar. The SFE process was carried out on
laboratory scale high-pressure extraction plant (HPEP, NOVA, Swiss,
Efferikon, Switzerland) described in detail by
Pekić et al. (1995)
. The
extraction process was carried out, and extraction yield was measured
after 15, 30, 60, 90, 120, 180, 240, 300 and 360 min of extraction, in
order to study the dynamics and kinetics of the process. The extraction
was stopped after it went into the diffusion-limited process.
2.2.4. Soxhlet extraction (SE)
Five grams of root samples were separately extracted with ethanol
(96%, w/w) in a Soxhlet apparatus for 6 h.
2.2.5. Ultrasonication-assisted extraction (UAE)
The powdered roots (2 g) was extracted with 50 mL of ethanol
(96%, w/w) for 60 min in a sonication bath at 30 °C.
The obtained extracts were concentrated under vacuum at 40 °C by
using a rotary vacuum evaporator. All samples were stored at 4 °C in the
dark until use.
2.3. Pro
file of bioactive compounds
Concerning our previous studies (
Uysal et al., 2017
), the total
amount of phenolics (TPC) (by standard Folin-Ciocalteu method) and
flavonoids (TFC) (by AlCl3
method) were determined. The
final results
were expressed as equivalents of standard compounds (gallic acid (mg
GAE/g) for TPC and rutin (mg RE/g) for TFC), respectively).
All HPLC
–MS/MS experiments were carried out on Dionex Ultimate
3000RS UHPLC instrument from Thermo Scientific (USA). The
chro-matographic runs were conducted at the temperature 25 ± 1 °C using a
Thermo Accucore C18
(100 mm x 2.1, mm i. d., 2.6
μm) column. The
mobile phase was composed of water acidi
fied with 0.1% formic acid
“A” and acetonitrile with 0.1% formic acid “B”. Mass spectrometry was
performed on Thermo Q-Exactive Orbitrap mass spectrometer (Thermo
Scienti
fic, USA) equipped with electrospray ionization probe interface
in positive and negative-ion mode. For collection and analyzing data
Thermo Scienti
fic Xcalibur 3.1 and TraceFinder Clinical Research 3.3
softwares (Thermo Scienti
fic, USA) were used. Compounds were
iden-tified by their exact molecular masses, isotopic patterns and
fragmen-tations. In the cases of fragmentations an own database was used. All
analytical details were given in supplementary material (
Zengin et al.,
2018c
).
2.4. Determination of antioxidant and enzyme inhibitory e
ffects
The root extracts were tested as sources of enzyme inhibitors on
some enzymes, including
α-amylase, α-glucosidase, cholinesterases,
and tyrosinase. The procedures of these assays are as reported in our
earlier work (
Uysal et al., 2017
). The enzyme inhibitor effects were
evaluated as equivalents of acarbose (for
α-amylase and α-glucosidase),
galantamine (for AChE and BChE), and kojic acid (for tyrosinase).
Different experiments spectrophotometrically screened antioxidant
capacity of the root extracts as phosphomolybdenum, quenching of
radicals (DPPH and ABTS), reduction potentials (FRAP and CUPRAC),
and ferrous ion chelating. The
findings were expressed as standard
compounds equivalents (mg TE/g and mg EDTAE/g). The procedures of
assays were given reported in our earlier work (
Uysal et al., 2017
).
2.5. Statistical analysis
One-way ANOVA was done to determine any di
fferences between
the different extraction methods following by Tukey’s test. p < 0.05
were assigned to be statistically significant. The heat map and Pearson
linear correlation were employed to recognize any relationship between
phytochemical contents and the observed biological activities. Besides,
principal component (PCA) and partial least squares discriminant
analysis (PLS-DA) analysis were performed to classify the performed
extraction methods. The statistical procedures were performed by R
software v. 3.5.1.
3. Results and discussion
The last decades have witnessed a renewed interest in
phytochem-icals for the pharmaceutical and nutraceutical industries (
Yahya et al.,
phytochemicals for the management of a plethora of human ailments.
Along with conventional extraction techniques such as maceration,
percolation, and Soxhlet extraction, newer enhanced techniques have
been developed to boost extraction of bioactive constituents from
plants. Non-conventional methods having the particularity of producing
higher extraction yield, being less destructive to extracted compounds,
and being environmentally friendly, have gained increasing importance
in drug discovery (
Saha et al., 2018
). In the present study, we have
attempted to elucidate the possible effect of different extraction
tech-niques on the biological activity of S. viridis roots.
Phytochemicals are chemical compounds, ubiquitously occurring in
plants, known to provide multiple health bene
fits such as anti-cancer,
antibacterial, antiviral, inflammatory, diabetic, and
anti-oxidant effects (
Guldiken et al., 2018
). Therefore, while determining
the possible bioactivity of plants, assessing the phytochemical
compo-sition is a crucial step. The amounts of phenolics and
flavonoids were
determined using well-known, rapid spectrophotometric methods,
namely Folin-Ciocalteu and Aluminum chloride tests, respectively.
From
Fig. 1
, the phenolic and
flavonoid contents of S. viridis ethanolic
root extracts followed this order: UAE > MAC > SE > MAE > SFE.
The total phenolics content of the hydro-ethanolic extract of S. viridis
aerial part obtained by UAE (
Grzegorczyk-Karolak and Kiss, 2018
) was
comparable to that of S. viridis ethanolic root extract obtained by the
same extraction method (
Fig. 1
). S. viridis roots (102.03 mg GAE/g
extract) possessed higher phenolics content compared to S. fruticosa
roots (80 mg GAE/g extract) (
Boukhary et al., 2016
). As observed from
Fig. 1
, the extraction technique in
fluenced extractability of flavonoids,
and UAE obtained the highest
flavonoid content. The differences among
the extracts were influenced by the different phenomena which occur in
different techniques. As it can be seen cavitation phenomenon has the
most substantial impact on both phenols and
flavonoids extraction.
Conventional techniques (MAC and SE) were proven as a better choice
than modern MAE technique. The reason could be the time of
extrac-tions. However, among the SE, MAC and MAE the di
fferences in total
phenols and
flavonoids were not notable, but there was a remarkable
difference in extraction times. Namely, the MAC and SE were performed
24 and 6 h, respectively, while MAE was performed only 30 min. Also,
the reason for lower content could be possible degradation during the
microwave irradiation. Data from the literature suggests that the
con-tent of polyphenolic compounds from di
fferent matrices increase up to
certain levels and then decreases as a consequence of microwave
irra-diation (
Ameer et al., 2017
). Moreover, too high microwave irradiation
of plant material could also lead to degradation of polyphenols (
Švarc-Gajić et al., 2013
). This implies that MAE has the potential to be used
for polyphenols extraction, but the extraction conditions, in the
first
place: the power of irradiation, and time should be optimized. As it was
expected, the SFE, which is suitable for non-polar components, offers
the lowers yield of polyphenolic compounds. Besides, previous studies
have indicated that UAE optimised
flavonoid extraction (
Zhang et al.,
2011
;
Zheng et al., 2016
).
HPLC
–MS/MS screening was applied to S. viridis ethanolic root
ex-tracts obtained by di
fferent extraction methods. Compounds
identifi-cation involved accurate molecular mass measurements (MS) and
ob-taining tandem mass spectra (MS/MS) followed by comparison with
database, literature and retention time of the standards. A total of 76
compounds were identified in the extracts obtained by SE, UAE, and
MAC, 73 compounds for MAE and 24 compounds for SFE (
Tables 1–5
;
Figure S1-S5). The selective identi
fication of compounds was provided
by SFE extraction. The results of all extracts showed that the main
components belong to salvianolic acids, polyphenols,
flavonoids, and
terpenoids. The presence of salvianolic acids was also reported in other
Salvia species such as S. miltiorrhiza (
Li et al., 2009
), S. przewalski
(salvianolic acid B and K) (
Li et al., 2013
;
Ożarowski et al., 2017
), S.
chinensis (salvianolic acid B, D) (
Li et al., 2013
), S. o
fficinalis (salvianolic
acid L) (
Lu and Foo, 2001
), S.
flava (salvianolic acid J) (
Ai et al., 1994
).
Diterpenes namely, 1-oxomicrostegiol, viroxicin, and 3-oxomicrostegiol
(m/z 313.18) were identi
fied in all extracts. Viridoquinone (m/z
297.18) was found in all extracts except UAE extract and similar
finding
was reported in S. viridis roots (
Rungsimakan and Rowan, 2014
). Two
unknown terpenoid isomers with m/z 485.36 were found in SFE
ex-tracts. Monoterpene glycoside lipedoside A (m/z 607.20), characterized
by high antioxidant properties (
Chen et al., 2002
) and phenylpropanoid
glycoside leucosceptoside A (m/z 637.21), identified in SE, UAE, MAE
and MAC extracts, were also identi
fied in S. viridis shoots (
Grzegorczyk-Karolak and Kiss, 2018
). Kynurenic acid (m/z 190.0), previously
iso-lated from S. officinalis (
Turski et al., 2015
), was identified in MAE, SE,
and UAE extracts of S. viridis ethanolic root extracts. Vanillin (m/z
153.44), present in all studied extracts, was also identi
fied in S.
digi-taloides roots (
Wu and Chan, 2014
). Compounds such as caffeoylglucose
(m/z 341.08), chlorogenic acid (m/z 355.10), apigenin-O-glucuronide
(m/z 445.07), martynoside (m/z 651.22) were also isolated from S.
viridis shoots (
Grzegorczyk-Karolak and Kiss, 2018
). Coniferyl aldehyde
(m/z 179.07) was previously isolated from Salvia plebeia(
Weng and
Wang, 2000
).
The role of reactive oxygen species and free radicals in the
patho-genesis of several human ailments is well established. A substantial
body of evidence has demonstrated the ability of phytochemicals to
influence important cellular and molecular mechanisms related to
health problems, including oxidative-stress related ones (
Zengin et al.,
2019
). In the present study, the antioxidant activity of S. viridis extracts
was assessed using three different classes of antioxidant assays (radical
scavenging, reducing power, and metal chelating). The results are
present in
Table 6
. S. viridis ethanolic root extract obtained by UAE
showed highest radical scavenging (240.00 and 302.85 mg TE/g for
DPPH and ABTS assays, respectively) and reducing (970.74, 704.27 mg
TE/g, and 2.84 mmol TE/g for CUPRAC, FRAP, and
phosphomo-lybdenum assays, respectively) activities. UAE carry some advantages;
namely use of low temperature preserving the integrity of
phyto-chemicals, short extraction time, increased extraction yield, making it
one of the most exciting techniques for the extraction of bioactive
components from plant materials (
Liu et al., 2018
). The influence of the
extraction technique on the antioxidant activity was already proven and
detailed explained in the literature (
Cvetanovi
ć et al., 2015
). The
an-tioxidant results were also correlated total bioactive components
namely total phenolics and
flavonoids content (
Fig. 2
A).
In contradiction with the aforementioned antioxidant assays, the
extract obtained by MAE (22.71 mg EDTAE/g) showed the most potent
metal chelating activity. This
finding was supported by correlation data
Fig. 1. Total bioactive compounds of the Salvia viridis extracts (Values ex-pressed are means ± S.D. of three parallel measurements. MAE: Microwave
assisted extraction; MAC: Maceration, SFE: Supercriticalfluid extraction, SE:
Soxhlet extraction; UAE: Ultrasonic assisted extraction. GAE: Gallic acid equivalent; RE: Rutin equivalent. Different letters indicate differences in the tested extracts (p < 0.05)).
Table 1 The chemical composition of SFE from Salvia viridis roots. No. Rt, min Compound Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 1 * 15,68 Vanillin C8 H8 O3 153,05517 125,0601 111,0445 93,0341 65,0394 2 16,83 Ethyl syringate C11 H14 O5 227,09195 181,0500 155,0706 140,0471 123,0444 95,0498 3 17,39 Syringaldehyde (3,5-Dimethoxy-4-hydroxybenzaldehyde) C9 H10 O4 183,06574 155,0705 140,0471 123,0444 95,0498 4 18,04 Antiarol (3,4,5-Trimethoxyphenol) C9 H12 O4 185,08139 170,0575 154,0626 153,0548 139,0392 125,0600 5 19,09 Indole-4-carbaldehyde C9 H7 NO 146,06059 118,0656 117,0575 91,0547 6 19,89 Coumarin C9 H6 O2 147,04461 119,0493 103,0548 91,0548 65,0393 7 20,11 Coniferyl aldehyde (4-Hydroxy-3-methoxycinnamaldehyde) C10 H10 O3 179,07082 161,0600 147,0443 133,0651 119,0496 105,0705 8 20,19 N-(2-Phenylethyl)acetamide C10 H13 NO 164,10754 122,0968 105,0705 103,0549 90,9484 79,0550 9 20,54 Sinapyl aldehyde (3,5-Dimethoxy-4-hydroxycinnamaldehyde) C11 H12 O4 209,08139 191,0708 177,0551 149,0601 145,0288 55,0187 10 30,50 Dimethoxy-trihydroxy(iso) fl avone isomer 1 C17 H14 O7 329,06613 314,1530 271,0245 11 34,35 Dihydroxy-dimethoxy(iso) fl avone C17 H14 O6 313,07122 298,0487 283,0254 255,0299 12 34,46 Dimethoxy-trihydroxy(iso) fl avone isomer 2 C17 H14 O7 329,06613 314,1533 271,0254 13 34,47 Genkwanin C1 6 H12 O5 283,06120 268,0380 239,0357 117,0337 14 34,91 Dihydroxy-trimethoxy(iso) fl avone C18 H16 O7 343,08178 328,0590 313,0359 298,0122 15 35,85 Hydroxy-trimethoxy(iso) fl avone C18 H16 O6 329,10252 314,0789 313,0711 300,0634 285,0764 16 36,52 Hydroxy-tetramethoxy(iso) fl avone C19 H18 O7 359,11308 344,0895 343,0816 329,0660 313,0703 17 36,65 1-Oxomicrostegiol C20 H24 O3 313,18037 295,1697 280,1465 243,1019 ( Rungsimakan and Rowan, 2014 ) 18 36,96 Viroxocin C20 H24 O3 313,18037 295,1696 227,1069 199,0757 ( Rungsimakan and Rowan, 2014 ) 19 38,18 Apigenin-4',7-dimethyl ether (4',7-Dimethoxy-5-hydroxy fl avone) C17 H14 O5 299,09195 284,0683 256,0735 20 38,62 3-Oxomicrostegiol C20 H24 O3 313,18037 295,1696 280,1457 243,1019 ( Rungsimakan and Rowan, 2014 ) 21 40,36 Hexadecanedioic acid C16 H30 O4 285,20659 267,1970 223,2067 22 41,81 Viridoquinone C20 H24 O2 297,18546 279,1747 269,1912 239,1433 ( Rungsimakan and Rowan, 2014 ) 23 49,39 Unknown terpenoid isomer 1 C31 H48 O4 485,36309 453,3677 425,3792 407,3676 271,2418 259,2427 24 49,84 Unknown terpenoid isomer 2 C31 H48 O4 485,36309 453,3720 425,3787 407,3680 271,2428 259,2424 * Con fi rmed by standard.
Table 2 The chemical composition of MAE from Salvia viridis roots. No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 1 1,24 Quinic acid C7 H12 O6 191,05557 173,0449 171,0288 127,0385 111,0440 85,0280 2 * 2,34 Gallic acid (3,4,5-Trihydroxybenzoic acid) C7 H6 O5 169,01370 125,0232 97,0283 81,0331 69,0331 3 12,69 Ca ff eoylglucose isomer 1 C15 H18 O9 341,08726 281,0675 251,0563 221,0453 179,0343 161,0235 ( Grzegorczyk-Karolak and Kiss, 2018 ) 4 13,90 Kynurenic acid C10 H7 NO 3 190,05042 162,0552 144,0449 116,0498 89,0393 5 14,24 Ca ff eoylglucose isomer 2 C15 H18 O9 341,08726 281,0672 251,0561 221,0454 179,0343 161,0235 ( Grzegorczyk-Karolak and Kiss, 2018 ) 6 * 14,27 Chlorogenic acid (3-O-Ca ff eoylquinic acid) C1 6 H18 O9 355,10291 163,0391 145,0286 135,0443 117,0338 107,0496 ( Grzegorczyk-Karolak and Kiss, 2018 ) 7 14,32 Ca ff eic acid C9 H8 O4 179,03444 135,0440 107,0487 8 15,19 Coumaroylhexose isomer 1 C15 H18 O8 325,09235 265,0721 235,0611 205,0503 163,0391 145,0285 9 * 15,54 Vanillin C8 H8 O3 153,05517 125,0600 111,0444 93,0340 65,0393 10 16,58 Coumaroylhexose isomer 2 C15 H18 O8 325,09235 265,0722 235,0612 205,0505 163,0392 145,0285 11 16,75 Feruloylhexose isomer 1 C16 H20 O9 355,10291 295,0831 235,0610 193,0503 175,0391 160,0155 12 16,76 Ethyl syringate C11 H14 O5 227,09195 181,0499 155,0705 140,0470 123,0444 95,0497 13 16,90 Coumaroylquinic acid C16 H18 O8 337,09235 191,0557 173,0446 163,0389 119,0490 93,0332 14 17,26 Ethyl gallate C9 H10 O5 197,04500 169,0133 125,0231 15 17,30 Syringaldehyde (3,5-Dimethoxy-4-hydroxybenzaldehyde) C9 H10 O4 183,06574 155,0704 140,0471 123,0444 95,0497 16 * 17,75 4-Coumaric acid C9 H8 O3 163,03952 119,0490 93,0331 17 17,88 Feruloylhexose isomer 2 C16 H20 O9 355,10291 295,0828 235,0612 193,0502 175,0391 160,0157 18 17,96 Antiarol (3,4,5-Trimethoxyphenol) C9 H12 O4 185,08139 170,0576 154,0626 153,0549 139,0392 125,0600 19 17,97 Ca ff eoylshikimic acid C16 H16 O8 335,07670 179,0343 161,0234 135,0441 20 18,09 Sinapoylhexose C17 H22 O10 385,11348 267,0725 249,0619 223,0610 208,0373 164,0469 21 18,99 Indole-4-carbaldehyde C9 H7 NO 146,06059 118,0655 117,0578 91,0548 22 19,05 Luteolin-C-hexoside-C-pentoside isomer 1 C26 H28 O15 579,13500 489,1022 459,0893 399,0728 369,0631 339,0513 23 19,12 Dimethoxy-hydroxybenzoic acid C9 H10 O5 197,04500 182,0215 166,9977 153,0547 138,0311 121,0282 24 19,32 Ferulic acid C10 H10 O4 193,05009 178,0264 149,0598 137,0232 134,0362 25 19,45 Luteolin-C-hexoside-C-pentoside isomer 2 C26 H28 O15 579,13500 489,1036 459,0953 399,0730 369,0621 339,0520 26 20,02 Coniferyl aldehyde (4-Hydroxy-3-methoxycinnamaldehyde) C10 H10 O3 179,07082 161,0598 147,0442 133,0651 119,0496 105,0705 27 20,15 N-(2-Phenylethyl)acetamide C10 H13 NO 164,10754 122,0967 105,0704 103,0549 90,9483 79,0553 28 20,22 Luteolin-C-hexoside-O-pentoside C26 H28 O15 581,15065 449,1086 431,0975 413,0873 329,0657 299,0553 29 20,37 Coumaroylshikimate C16 H16 O7 319,08178 163,0392 155,0338 119,0490 30 20,47 Sinapyl aldehyde (3,5-Dimethoxy-4-hydroxycinnamaldehyde) C11 H12 O4 209,08139 191,0706 177,0548 149,0599 145,0286 55,0186 31 20,83 Myricetin-O-hexoside C21 H20 O13 479,08257 317,0314 316,0232 287,0200 271,0246 32 21,72 Apigenin-C-hexoside-O-pentoside C26 H28 O14 565,15574 433,1133 415,1028 337,0708 313,0707 283,0602 33 21,98 Verbascoside C29 H36 O15 623,19760 461,1680 315,1084 161,0234 133,0284 ( Grzegorczyk-Karolak and Kiss, 2018 ) 34 22,11 Luteolin-O-(pentosyl)hexoside C26 H28 O15 581,15065 449,1086 287,0553 35 22,20 Luteolin-O-glucuronide C21 H18 O12 461,07201 285,0411 217,0502 199,0400 151,0027 133,0282 36 22,35 Luteolin-7-O-glucoside (Cynaroside) C21 H20 O11 447,09274 327,0504 285,0411 284,0333 256,0376 151,0027 37 22,40 Luteolin-O-(deoxyhexosyl)hexoside C27 H30 O15 595,16630 449,1083 287,0552 38 * 22,90 Isoquercitrin (Hirsutrin, Quercetin-3-O-glucoside) C21 H20 O12 463,08765 301,0360 300,0280 271,0254 255,0302 39 23,13 3-[(1-Carboxyvinyl)oxy]benzoic acid C10 H8 O5 207,02935 137,0233 135,0441 93,0331 87,0073 40 23,14 Rosmarinic acid-O-hexoside C24 H26 O13 521,12952 359,0774 341,0885 323,0777 197,0450 161,0234 41 23,23 Salvianolic acid isomer C36 H30 O16 717,14557 519,0942 339,0513 321,0415 295,0615 277,0507 42 23,31 Lipedoside A isomer 1 C29 H36 O14 607,20269 461,1669 443,0985 297,0403 161,0234 135,0441 ( Grzegorczyk-Karolak and Kiss, 2018 ) 43 23,59 Lipedoside A isomer 2 C29 H36 O14 607,20269 461,1667 443,0992 297,0400 161,0234 135,0440 ( Grzegorczyk-Karolak and Kiss, 2018 ) 44 23,64 Leucosceptoside A C30 H38 O15 637,21325 461,1677 315,1095 193,0503 175,0392 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 45 23,89 Apigenin-O-(deoxyhexosyl)hexoside C27 H30 O14 577,15574 433,1135 271,0603 46 * 23,95 Cosmosiin (Apigetrin, Apigenin-7-O-glucoside) C21 H20 O10 433,11347 271,0603 153,0181 119,0499 47 24,09 Apigenin-O-glucuronide C21 H18 O11 445,07709 269,0460 175,0241 113,0231 ( Grzegorczyk-Karolak and Kiss, 2018 ) 48 24,34 Methoxy-trihydroxy fl avone-O-glucuronide C22 H20 O12 477,10331 301,0710 286,0476 258,0520 49 24,40 Methyl ca ff eate C10 H10 O4 195,06574 163,0391 145,0286 135,0443 117,0339 107,0496 50 24,41 Rosmarinic acid (Labiatenic acid) C18 H16 O8 359,07670 197,0451 179,0343 161,0234 135,0440 123,0439 ( Grzegorczyk-Karolak and Kiss, 2018 ) 51 24,65 N-trans-Feruloyltyramine C18 H19 NO 4 314,13924 177,0549 145,0286 121,0651 52 25,85 Martynoside C31 H40 O15 651,22890 475,1833 193,0501 175,0391 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 53 25,88 Ethyl ca ff eate C11 H12 O4 209,08139 163,0392 145,0286 135,0444 117,0339 107,0860 54 26,28 3-O-Methylrosmarinic acid C19 H18 O8 373,09235 197,0451 179,0342 175,0391 160,0156 135,0440 (continued on next page )
Table 2 (continued ) No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 55 26,97 Pentahydroxy fl avone C15 H10 O7 301,03483 273,0393 178,9984 151,0027 107,0123 56 * 27,17 Naringenin C15 H12 O5 271,06065 177,0183 165,0185 151,0026 119,0489 107,0126 57 * 27,85 Luteolin (3',4',5,7-Tetrahydroxy fl avone) C15 H10 O6 285,03991 217,0503 199,0400 151,0027 133,0283 107,0127 58 27,88 Di-O-methylellagic acid C16 H10 O8 329,02975 314,0075 298,9842 270,9896 59 28,54 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 1 C17 H14 O6 315,08687 205,0498 163,0391 153,0545 145,0286 135,0444 60 29,28 Luteolin-O-(coumaroyl)hexoside C30 H26 O13 593,12952 285,0410 284,0329 145,0282 61 29,54 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 2 C17 H14 O6 315,08687 205,0500 163,0391 153,0549 145,0285 135,0443 62 * 29,69 Apigenin (4',5,7-Trihydroxy fl avone) C15 H10 O5 269,04500 227,0353 225,0559 201,0554 151,0026 117,0332 63 29,90 Chrysoeriol (Scoparol, 3'-Methoxy-4',5,7-trihydroxy fl avone) C16 H12 O6 299,05556 284,0331 256,0378 227,0344 151,0024 107,0119 64 30,23 Tri-O-methylellagic acid C17 H12 O8 343,04540 328,0226 312,9996 297,9760 65 30,58 Dimethoxy-trihydroxy(iso) fl avone C17 H14 O7 329,06613 314,1529 271,0252 66 32,55 Methoxy-trihydroxy fl avone C16 H12 O6 299,05556 284,0331 256,0372 133,0284 67 34,54 Genkwanin C16 H12 O5 283,06120 268,0382 239,0355 117,0328 68 35,83 Hydroxy-trimethoxy(iso) fl avone C18 H16 O6 329,10252 314,0793 313,0710 300,0632 285,0759 69 36,63 1-Oxomicrostegiol C20 H24 O3 313,18037 295,1692 280,1461 243,1018 ( Rungsimakan and Rowan, 2014 ) 70 36,95 Viroxocin C20 H24 O3 313,18037 295,1694 227,1068 199,0756 ( Rungsimakan and Rowan, 2014 ) 71 38,62 3-Oxomicrostegiol C20 H24 O3 313,18037 295,1698 280,1465 243,1019 ( Rungsimakan and Rowan, 2014 ) 72 40,36 Hexadecanedioic acid C16 H30 O4 285,20659 267,1970 223,2067 73 41,81 Viridoquinone C20 H24 O2 297,18546 279,1748 269,1916 239,1432 ( Rungsimakan and Rowan, 2014 ) * Con fi rmed by standard.
Table 3 The chemical composition of SE from Salvia viridis roots. No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 1 1,24 Quinic acid C7 H12 O6 191,05557 173,0444 171,0286 127,0388 111,0439 85,0280 2 * 2,38 Gallic acid (3,4,5-Trihydroxybenzoic acid) C7 H6 O5 169,01370 125,0232 97,0282 81,0331 69,0331 3 12,82 Ca ff eoylglucose isomer 1 C15 H18 O9 341,08726 281,0671 251,0564 221,0456 179,0343 161,0234 ( Grzegorczyk-Karolak and Kiss, 2018 ) 4 13,92 Kynurenic acid C10 H7 NO 3 190,05042 162,0552 144,0444 116,0500 89,0390 5 14,31 Ca ff eoylglucose isomer 2 C15 H18 O9 341,08726 281,0672 251,0563 221,0453 179,0343 161,0234 ( Grzegorczyk-Karolak and Kiss, 2018 ) 6 * 14,33 Chlorogenic acid (3-O-Ca ff eoylquinic acid) C16 H18 O9 355,10291 163,0392 145,0287 135,0445 117,0339 107,0496 ( Grzegorczyk-Karolak and Kiss, 2018 ) 7 14,43 Ca ff eic acid C9 H8 O4 179,03444 135,0441 107,0488 8 15,29 Coumaroylhexose isomer 1 C15 H18 O8 325,09235 265,0721 235,0611 205,0503 163,0391 145,0285 9 * 15,60 Vanillin C8 H8 O3 153,05517 125,0601 111,0445 93,0341 65,0394 10 16,66 Coumaroylhexose isomer 2 C15 H18 O8 325,09235 265,0721 235,0611 205,0504 163,0391 145,0285 11 16,78 Ethyl syringate C11 H14 O5 227,09195 181,0499 155,0705 140,0471 123,0445 95,0496 12 16,82 Feruloylhexose isomer 1 C16 H20 O9 355,10291 295,0829 235,0609 193,0503 175,0391 160,0155 13 16,96 Coumaroylquinic acid C16 H18 O8 337,09235 191,0557 173,0447 163,0394 119,0492 93,0332 14 17,34 Ethyl gallate C9 H10 O5 197,04500 169,0134 125,0231 15 17,37 Syringaldehyde (3,5-Dimethoxy-4-hydroxybenzaldehyde) C9 H10 O4 183,06574 155,0705 140,0471 123,0444 95,0498 16 * 17,85 4-Coumaric acid C9 H8 O3 163,03952 119,0489 93,0332 17 17,94 Feruloylhexose isomer 2 C16 H20 O9 355,10291 295,0832 235,0612 193,0502 175,0392 160,0155 18 18,00 Antiarol (3,4,5-Trimethoxyphenol) C9 H12 O4 185,08139 170,0575 154,0626 153,0549 139,0392 125,0601 19 18,05 Ca ff eoylshikimic acid C16 H16 O8 335,07670 179,0342 161,0234 135,0441 20 18,12 Sinapoylhexose C17 H22 O10 385,11348 267,0725 249,0619 223,0608 208,0371 164,0469 21 19,05 Indole-4-carbaldehyde C9 H7 NO 146,06059 118,0655 117,0578 91,0548 22 19,10 Luteolin-C-hexoside-C-pentoside isomer 1 C26 H28 O15 579,13500 489,1051 459,0941 399,0740 369,0625 339,0499 23 19,12 Dimethoxy-hydroxybenzoic acid C9 H10 O5 197,04500 182,0215 166,9977 153,0547 138,0310 121,0282 24 19,40 Ferulic acid C10 H10 O4 193,05009 178,0264 149,0597 137,0233 134,0362 25 19,50 Luteolin-C-hexoside-C-pentoside isomer 2 C26 H28 O15 579,13500 489,1029 459,0933 399,0727 369,0619 339,0503 26 20,06 Coniferyl aldehyde (4-Hydroxy-3-methoxycinnamaldehyde) C10 H10 O3 179,07082 161,0599 147,0442 133,0652 119,0495 105,0704 27 20,19 N-(2-Phenylethyl)acetamide C10 H13 NO 164,10754 122,0969 105,0705 103,0549 90,9483 28 20,28 Luteolin-C-hexoside-O-pentoside C26 H28 O15 581,15065 449,1087 431,0979 413,0873 329,0660 299,0554 29 20,45 Coumaroylshikimate C16 H16 O7 319,08178 163,0391 155,0340 119,0490 30 20,51 Sinapyl aldehyde (3,5-Dimethoxy-4-hydroxycinnamaldehyde) C11 H12 O4 209,08139 191,0708 177,0550 149,0600 145,0287 55,0187 31 20,91 Myricetin-O-hexoside C21 H20 O13 479,08257 317,0306 316,0225 287,0193 271,0253 32 21,77 Apigenin-C-hexoside-O-pentoside C26 H28 O14 565,15574 433,1137 415,1030 337,0712 313,0711 283,0605 33 22,03 Verbascoside C29 H36 O15 623,19760 461,1679 315,1077 161,0234 133,0284 ( Grzegorczyk-Karolak and Kiss, 2018 ) 34 22,11 Luteolin-O-(pentosyl)hexoside C26 H28 O15 581,15065 449,1087 287,0553 35 22,35 Luteolin-7-O-glucoside (Cynaroside) C21 H20 O11 447,09274 327,0504 285,0411 284,0333 256,0376 151,0027 36 22,39 Luteolin-O-glucuronide C21 H18 O12 461,07201 285,0411 217,0496 199,0397 151,0026 133,0281 37 22,44 Luteolin-O-(deoxyhexosyl)hexoside C27 H30 O15 595,16630 449,1084 287,0552 38 * 22,97 Isoquercitrin (Hirsutrin, Quercetin-3-O-glucoside) C21 H20 O12 463,08765 301,0360 300,0280 271,0253 255,0303 39 23,15 3-[(1-Carboxyvinyl)oxy]benzoic acid C10 H8 O5 207,02935 137,0234 135,0441 93,0332 87,0073 40 23,18 Rosmarinic acid-O-hexoside C24 H26 O13 521,12952 359,0776 341,0942 323,0779 197,0451 161,0234 41 23,27 Salvianolic acid isomer C36 H30 O16 717,14557 519,0959 339,0506 321,0411 295,0616 277,0492 42 23,36 Lipedoside A isomer 1 C29 H36 O14 607,20269 461,1671 443,0986 297,0404 161,0234 135,0443 ( Grzegorczyk-Karolak and Kiss, 2018 ) 43 23,64 Lipedoside A isomer 2 C29 H36 O14 607,20269 461,1679 443,1014 297,0414 161,0235 135,0442 ( Grzegorczyk-Karolak and Kiss, 2018 ) 44 23,69 Leucosceptoside A C30 H38 O15 637,21325 461,1675 315,1102 193,0502 175,0392 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 45 23,95 Apigenin-O-(deoxyhexosyl)hexoside C27 H30 O14 579,17139 433,1135 271,0604 46 * 23,99 Cosmosiin (Apigetrin, Apigenin-7-O-glucoside) C21 H20 O10 433,11347 271,0604 153,0181 119,0496 47 24,15 Apigenin-O-glucuronide C21 H18 O11 445,07709 269,0460 175,0239 113,0231 ( Grzegorczyk-Karolak and Kiss, 2018 ) 48 24,36 Methoxy-trihydroxy fl avone-O-glucuronide C22 H20 O12 477,10331 301,0710 286,0476 258,0520 49 24,42 Methyl ca ff eate C10 H10 O4 195,06574 163,0392 145,0287 135,0444 117,0340 107,0496 50 24,43 Rosmarinic acid (Labiatenic acid) C18 H16 O8 359,07670 197,0452 179,0344 161,0235 135,0441 123,0439 ( Grzegorczyk-Karolak and Kiss, 2018 ) 51 24,70 N-trans-Feruloyltyramine C1 8 H19 NO 4 314,13924 177,0549 145,0287 121,0652 52 25,76 3-O-Methylellagic acid C15 H8 O8 315,01410 299,9917 53 25,90 Martynoside C31 H40 O15 651,22890 475,1844 193,0502 175,0392 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 54 25,93 Ethyl ca ff eate C11 H12 O4 209,08139 163,0392 145,0287 135,0445 117,0339 107,0858 (continued on next page )
Table 3 (continued ) No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 55 26,31 3-O-Methylrosmarinic acid C19 H18 O8 373,09235 197,0451 179,0342 175,0392 160,0156 135,0441 56 27,02 Pentahydroxy fl avone C15 H10 O7 301,03483 273,0424 178,9978 151,0028 107,0124 57 * 27,23 Naringenin C15 H12 O5 271,06065 177,0186 165,0179 151,0027 119,0490 107,0126 58 * 27,91 Luteolin (3',4',5,7-Tetrahydroxy fl avone) C15 H10 O6 285,03991 217,0501 199,0398 151,0027 133,0283 107,0125 59 27,95 Di-O-methylellagic acid C16 H10 O8 329,02975 314,0073 298,9839 270,9883 60 28,59 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 1 C17 H14 O6 315,08687 205,0498 163,0392 153,0549 145,0286 135,0444 61 29,33 Luteolin-O-(coumaroyl)hexoside C30 H26 O13 593,12952 285,0411 284,0337 145,0286 62 29,59 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 2 C17 H14 O6 315,08687 205,0498 163,0392 153,0549 145,0287 135,0444 63 * 29,73 Apigenin (4',5,7-Trihydroxy fl avone) C15 H10 O5 269,04500 227,0342 225,0555 201,0557 151,0027 117,0332 64 29,97 Chrysoeriol (Scoparol, 3'-Methoxy-4',5,7-trihydroxy fl avone) C16 H12 O6 299,05556 284,0332 256,0380 227,0345 151,0027 107,0125 65 30,27 Tri-O-methylellagic acid C17 H12 O8 343,04540 328,0227 312,9999 297,9759 66 30,63 Dimethoxy-trihydroxy(iso) fl avone C17 H14 O7 329,06613 314,1530 271,0252 67 32,58 Methoxy-trihydroxy fl avone C16 H12 O6 299,05556 284,0333 256,0376 133,0277 68 34,46 Dihydroxy-dimethoxy(iso) fl avone C17 H14 O6 313,07122 298,0488 283,0253 255,0301 69 34,59 Genkwanin C16 H12 O5 283,06120 268,0382 239,0347 117,0334 70 35,84 Hydroxy-trimethoxy(iso) fl avone C18 H16 O6 329,10252 314,0787 313,0710 300,0631 285,0760 71 36,52 Hydroxy-tetramethoxy(iso) fl avone C19 H18 O7 359,11308 344,0893 343,0814 329,0661 313,0736 72 36,63 1-Oxomicrostegiol C20 H24 O3 313,18037 295,1694 280,1461 243,1017 ( Rungsimakan and Rowan, 2014 ) 73 36,95 Viroxocin C20 H24 O3 313,18037 295,1693 227,1069 199,0757 ( Rungsimakan and Rowan, 2014 ) 74 38,61 3-Oxomicrostegiol C20 H24 O3 313,18037 295,1696 280,1462 243,1018 ( Rungsimakan and Rowan, 2014 ) 75 40,38 Hexadecanedioic acid C16 H30 O4 285,20659 267,1975 223,2063 76 41,81 Viridoquinone C20 H24 O2 297,18546 279,1749 269,1901 239,1433 ( Rungsimakan and Rowan, 2014 ) * Con fi rmed by standard.
Table 4 The chemical composition of UAE from Salvia viridis roots. No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 1 1,26 Quinic acid C7 H12 O6 191,05557 173,0446 171,0286 127,0388 111,0441 85,0281 2 * 2,42 Gallic acid (3,4,5-Trihydroxybenzoic acid) C7 H6 O5 169,01370 125,0232 97,0281 81,0333 69,0331 3 12,89 Ca ff eoylglucose isomer 1 C15 H18 O9 341,08726 281,0674 251,0564 221,0455 179,0342 161,0235 ( Grzegorczyk-Karolak and Kiss, 2018 ) 4 13,84 Kynurenic acid C10 H7 NO 3 190,05042 162,0552 144,0438 116,0502 89,0393 5 * 14,41 Chlorogenic acid (3-O-Ca ff eoylquinic acid) C16 H18 O9 355,10291 163,0392 145,0287 135,0444 117,0340 107,0496 ( Grzegorczyk-Karolak and Kiss, 2018 ) 6 14,49 Ca ff eoylglucose isomer 2 C15 H18 O9 341,08726 281,0674 251,0564 221,0454 179,0343 161,0234 ( Grzegorczyk-Karolak and Kiss, 2018 ) 7 14,54 Ca ff eic acid C9 H8 O4 179,03444 135,0441 107,0491 8 15,35 Coumaroylhexose isomer 1 C15 H18 O8 325,09235 265,0722 235,0611 205,0504 163,0392 145,0285 9 * 15,72 Vanillin C8 H8 O3 153,05517 125,0601 111,0445 93,0341 65,0394 10 16,73 Coumaroylhexose isomer 2 C15 H18 O8 325,09235 265,0722 235,0610 205,0504 163,0392 145,0285 11 16,86 Ethyl syringate C11 H14 O5 227,09195 181,0500 155,0705 140,0471 123,0445 95,0495 12 16,87 Feruloylhexose isomer 1 C16 H20 O9 355,10291 295,0828 235,0609 193,0503 175,0392 160,0158 13 16,99 Coumaroylquinic acid C16 H18 O8 337,09235 191,0556 173,0448 163,0391 119,0488 93,0331 14 17,41 Ethyl gallate C9 H10 O5 197,04500 169,0134 125,0232 15 17,45 Syringaldehyde (3,5-Dimethoxy-4-hydroxybenzaldehyde) C9 H10 O4 183,06574 155,0706 140,0472 123,0445 95,0498 16 * 17,93 4-Coumaric acid C9 H8 O3 163,03952 119,0490 93,0332 17 17,98 Feruloylhexose isomer 2 C16 H20 O9 355,10291 295,0828 235,0609 193,0503 175,0392 160,0158 18 18,08 Antiarol (3,4,5-Trimethoxyphenol) C9 H12 O4 185,08139 170,0576 154,0627 153,0549 139,0394 125,0601 19 18,11 Ca ff eoylshikimic acid C16 H16 O8 335,07670 179,0343 161,0235 135,0441 20 18,13 Sinapoylhexose C17 H22 O10 385,11348 267,0725 249,0617 223,0611 208,0375 164,0469 21 19,09 Dimethoxy-hydroxybenzoic acid C9 H10 O5 197,04500 182,0215 166,9979 153,0547 138,0313 121,0283 22 19,15 Indole-4-carbaldehyde C9 H7 NO 146,06059 118,0656 117,0577 91,0548 23 19,16 Luteolin-C-hexoside-C-pentoside isomer 1 C26 H28 O15 579,13500 489,1054 459,0939 399,0729 369,0623 339,0517 24 19,44 Ferulic acid C10 H10 O4 193,05009 178,0264 149,0596 137,0231 134,0363 25 19,56 Luteolin-C-hexoside-C-pentoside isomer 2 C26 H28 O15 579,13500 489,1046 459,0942 399,0729 369,0623 339,0518 26 20,14 Coniferyl aldehyde (4-Hydroxy-3-methoxycinnamaldehyde) C10 H10 O3 179,07082 161,0600 147,0443 133,0652 119,0496 105,0704 27 20,25 N-(2-Phenylethyl)acetamide C10 H13 NO 164,10754 122,0969 105,0704 103,0547 90,9483 28 20,35 Luteolin-C-hexoside-O-pentoside C26 H28 O15 581,15065 449,1086 431,0985 413,0877 329,0660 299,0554 29 20,48 Coumaroylshikimate C16 H16 O7 319,08178 163,0392 155,0337 119,0489 30 20,58 Sinapyl aldehyde (3,5-Dimethoxy-4-hydroxycinnamaldehyde) C11 H12 O4 209,08139 191,0707 177,0549 149,0600 145,0286 55,0187 31 20,97 Myricetin-O-hexoside C21 H20 O13 479,08257 317,0299 316,0234 287,0194 271,0245 32 21,82 Apigenin-C-hexoside-O-pentoside C26 H28 O14 565,15574 433,1137 415,1032 337,0709 313,0711 283,0605 33 22,06 Verbascoside C29 H36 O15 623,19760 461,1672 315,1111 161,0235 133,0285 ( Grzegorczyk-Karolak and Kiss, 2018 ) 34 22,17 Luteolin-O-(pentosyl)hexoside C26 H28 O15 581,15065 449,1086 287,0554 35 22,41 Luteolin-O-glucuronide C21 H18 O12 461,07201 285,0410 217,0499 199,0399 151,0028 133,0281 36 22,45 Luteolin-7-O-glucoside (Cynaroside) C21 H20 O11 447,09274 327,0512 285,0411 284,0332 256,0381 151,0022 37 22,51 Luteolin-O-(deoxyhexosyl)hexoside C27 H30 O15 595,16630 449,1085 287,0553 38 22,72 Apigenin-C-hexoside-O-deoxyhexoside C27 H30 O14 579,17139 433,1136 415,1037 337,0709 313,0709 283,0602 39 * 23,02 Isoquercitrin (Hirsutrin, Quercetin-3-O-glucoside) C21 H20 O12 463,08765 301,0359 300,0281 271,0253 255,0301 40 23,11 3-[(1-Carboxyvinyl)oxy]benzoic acid C10 H8 O5 207,02935 137,0234 135,0441 93,0332 87,0073 41 23,19 Rosmarinic acid-O-hexoside C2 4H 26 O13 521,12952 359,0776 341,0901 323,0778 197,0452 161,0234 42 23,29 Salvianolic acid isomer C36 H30 O16 717,14557 519,0934 339,0526 321,0408 295,0616 277,0507 43 23,39 Lipedoside A isomer 1 C29 H36 O14 607,20269 461,1674 443,0991 297,0414 161,0235 135,0440 ( Grzegorczyk-Karolak and Kiss, 2018 ) 44 23,67 Lipedoside A isomer 2 C29 H36 O14 607,20269 461,1673 443,1001 297,0411 161,0235 135,0441 ( Grzegorczyk-Karolak and Kiss, 2018 ) 45 23,72 Leucosceptoside A C30 H38 O15 637,21325 461,1674 315,1088 193,0504 175,0392 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 46 24,01 Apigenin-O-(deoxyhexosyl)hexoside C27 H30 O14 579,17139 433,1136 271,0603 47 * 24,07 Cosmosiin (Apigetrin, Apigenin-7-O-glucoside) C21 H20 O10 433,11347 271,0603 153,0183 119,0495 48 24,15 Apigenin-O-glucuronide C21 H18 O11 445,07709 269,0460 175,0240 113,0231 ( Grzegorczyk-Karolak and Kiss, 2018 ) 49 24,41 Methoxy-trihydroxy fl avone-O-glucuronide C22 H20 O12 477,10331 301,0709 286,0476 258,0517 50 24,45 Methyl ca ff eate C10 H10 O4 195,06574 163,0392 145,0287 135,0444 117,0338 107,0500 51 24,46 Rosmarinic acid (Labiatenic acid) C18 H16 O8 359,07670 197,0452 179,0342 161,0234 135,0441 123,0439 ( Grzegorczyk-Karolak and Kiss, 2018 ) 52 24,75 N-trans-Feruloyltyramine C18 H19 NO 4 314,13924 177,0550 145,0287 121,0652 53 25,93 Martynoside C31 H40 O15 651,22890 475,1821 193,0501 175,0392 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 54 25,98 Ethyl ca ff eate C11 H12 O4 209,08139 163,0392 145,0287 135,0444 117,0339 107,0861 (continued on next page )
Table 4 (continued ) No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 55 26,34 3-O-Methylrosmarinic acid C19 H18 O8 373,09235 197,0452 179,0342 175,0392 160,0156 135,0440 56 27,09 Pentahydroxy fl avone C15 H10 O7 301,03483 273,0427 178,9980 151,0028 107,0126 57 * 27,28 Naringenin C15 H12 O5 271,06065 177,0178 165,0179 151,0028 119,0489 107,0122 58 * 27,95 Luteolin (3',4',5,7-Tetrahydroxy fl avone) C15 H10 O6 285,03991 217,0505 199,0399 151,0027 133,0283 107,0126 59 27,99 Di-O-methylellagic acid C16 H10 O8 329,02975 314,0078 298,9837 270,9889 60 28,64 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 1 C17 H14 O6 315,08687 205,0502 163,0392 153,0550 145,0286 135,0444 61 29,38 Luteolin-O-(coumaroyl)hexoside C30 H26 O13 593,12952 285,0411 284,0331 145,0286 62 29,64 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 2 C17 H14 O6 315,08687 205,0501 163,0392 153,0549 145,0287 135,0444 63 * 29,78 Apigenin (4',5,7-Trihydroxy fl avone) C15 H10 O5 269,04500 227,0342 225,0558 201,0551 151,0026 117,0331 64 30,00 Chrysoeriol (Scoparol, 3'-Methoxy-4',5,7-trihydroxy fl avone) C16 H12 O6 299,05556 284,0331 256,0378 227,0340 151,0025 107,0124 65 30,33 Tri-O-methylellagic acid C17 H12 O8 343,04540 328,0227 312,9999 297,9757 66 30,67 Dimethoxy-trihydroxy(iso) fl avone C17 H14 O7 329,06613 314,1531 271,0254 67 32,61 Methoxy-trihydroxy fl avone C16 H12 O6 299,05556 284,0331 256,0377 133,0281 68 34,49 Dihydroxy-dimethoxy(iso) fl avone C17 H14 O6 313,07122 298,0488 283,0255 255,0301 69 34,61 Genkwanin C16 H12 O5 283,06120 268,0382 239,0352 117,0328 70 35,90 Hydroxy-trimethoxy(iso) fl avone C18 H16 O6 329,10252 314,0789 313,0710 300,0633 285,0760 71 36,57 Hydroxy-tetramethoxy(iso) fl avone C19 H18 O7 359,11308 344,0895 343,0818 329,0662 313,0708 72 36,67 1-Oxomicrostegiol C20 H24 O3 313,18037 295,1696 280,1466 243,1019 ( Rungsimakan and Rowan, 2014 ) 73 36,99 Viroxocin C20 H24 O3 313,18037 295,1697 227,1069 199,0757 ( Rungsimakan and Rowan, 2014 ) 74 38,24 Apigenin-4',7-dimethyl ether (4',7-Dimethoxy-5-hydroxy fl avone) C17 H14 O5 299,09195 284,0685 256,0734 75 38,64 3-Oxomicrostegiol C20 H24 O3 313,18037 295,1697 280,1462 243,1019 ( Rungsimakan and Rowan, 2014 ) 76 40,40 Hexadecanedioic acid C16 H30 O4 285,20659 267,1972 223,2065 * Con fi rmed by standard.
Table 5 The chemical composition of MAC extract from Salvia viridis roots. No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 1 1,24 Quinic acid C7 H12 O6 191,05557 173,0446 171,0287 127,0391 111,0438 85,0280 2 * 2,36 Gallic acid (3,4,5-Trihydroxybenzoic acid) C7 H6 O5 169,01370 125,0232 97,0282 81,0334 69,0331 3 12,78 Ca ff eoylglucose isomer 1 C15 H18 O9 341,08726 281,0674 251,0566 221,0453 179,0344 161,0235 ( Grzegorczyk-Karolak and Kiss, 2018 ) 4 13,69 Kynurenic acid C10 H7 NO 3 190,05042 162,0552 144,0440 116,0499 89,0391 5 14,37 Ca ff eic acid C9 H8 O4 179,03444 135,0441 107,0489 6 14,37 Ca ff eoylglucose isomer 2 C15 H18 O9 341,08726 281,0673 251,0563 221,0453 179,0343 161,0234 ( Grzegorczyk-Karolak and Kiss, 2018 ) 7 * 14,43 Chlorogenic acid (3-O-Ca ff eoylquinic acid) C16 H18 O9 355,10291 163,0392 145,0287 135,0444 117,0339 107,0495 ( Grzegorczyk-Karolak and Kiss, 2018 ) 8 15,24 Coumaroylhexose isomer 1 C15 H18 O8 325,09235 265,0723 235,0612 205,0504 163,0392 145,0285 9 * 15,72 Vanillin C8 H8 O3 153,05517 125,0601 111,0445 93,0341 65,0394 10 16,62 Coumaroylhexose isomer 2 C15 H18 O8 325,09235 265,0721 235,0612 205,0504 163,0392 145,0285 11 16,76 Feruloylhexose isomer 1 C16 H20 O9 355,10291 295,0831 235,0612 193,0502 175,0393 160,0157 12 16,85 Ethyl syringate C11 H14 O5 227,09195 181,0499 155,0705 140,0471 123,0443 95,0497 13 16,87 Coumaroylquinic acid C16 H18 O8 337,09235 191,0557 173,0447 163,0394 119,0489 93,0332 14 17,27 Ethyl gallate C9 H10 O5 197,04500 169,0134 125,0232 15 17,46 Syringaldehyde (3,5-Dimethoxy-4-hydroxybenzaldehyde) C9 H10 O4 183,06574 155,0706 140,0472 123,0445 95,0498 16 * 17,78 4-Coumaric acid C9 H8 O3 163,03952 119,0490 93,0331 17 17,88 Feruloylhexose isomer 2 C16 H20 O9 355,10291 295,0829 235,0612 193,0503 175,0393 160,0156 18 17,95 Ca ff eoylshikimic acid C16 H16 O8 335,07670 179,0343 161,0234 135,0441 19 18,08 Antiarol (3,4,5-Trimethoxyphenol) C9 H12 O4 185,08139 170,0579 154,0627 153,0550 139,0393 125,0601 20 18,13 Sinapoylhexose C17 H22 O10 385,11348 267,0727 249,0616 223,0609 208,0376 164,0469 21 18,86 Dimethoxy-hydroxybenzoic acid C9 H10 O5 197,04500 182,0215 166,9977 153,0548 138,0314 121,0283 22 19,07 Luteolin-C-hexoside-C-pentoside isomer 1 C26 H28 O15 579,13500 489,1049 459,0924 399,0727 369,0621 339,0534 23 19,15 Indole-4-carbaldehyde C9 H7 NO 146,06059 118,0656 117,0580 91,0549 24 19,30 Ferulic acid C10 H10 O4 193,05009 178,0266 149,0598 137,0233 134,0363 25 19,46 Luteolin-C-hexoside-C-pentoside isomer 2 C26 H28 O15 579,13500 489,1044 459,0945 399,0730 369,0623 339,0517 26 20,16 Coniferyl aldehyde (4-Hydroxy-3-methoxycinnamaldehyde) C10 H10 O3 179,07082 161,0600 147,0444 133,0652 119,0496 105,0704 27 20,24 N-(2-Phenylethyl)acetamide C10 H13 NO 164,10754 122,0968 105,0705 103,0546 90,9484 28 20,37 Luteolin-C-hexoside-O-pentoside C26 H28 O15 581,15065 449,1086 431,0980 413,0877 329,0661 299,0554 29 20,39 Coumaroylshikimate C16 H16 O7 319,08178 163,0392 155,0343 119,0494 30 20,58 Sinapyl aldehyde (3,5-Dimethoxy-4-hydroxycinnamaldehyde) C11 H12 O4 209,08139 191,0706 177,0550 149,0600 145,0287 55,0187 31 20,87 Myricetin-O-hexoside C21 H20 O13 479,08257 317,0311 316,0229 287,0201 271,0250 32 21,85 Apigenin-C-hexoside-O-pentoside C26 H28 O14 565,15574 433,1134 415,1028 337,0710 313,0710 283,0604 33 21,98 Verbascoside C29 H36 O15 623,19760 461,1671 315,1102 161,0235 133,0285 ( Grzegorczyk-Karolak and Kiss, 2018 ) 34 22,21 Luteolin-O-(pentosyl)hexoside C26 H28 O15 581,15065 449,1088 287,0554 35 22,30 Luteolin-O-glucuronide C21 H18 O12 461,07201 285,0412 217,0502 199,0402 151,0026 133,0280 36 22,36 Luteolin-7-O-glucoside (Cynaroside) C21 H20 O11 447,09274 327,0530 285,0412 284,0333 256,0382 151,0028 37 22,53 Luteolin-O-(deoxyhexosyl)hexoside C27 H30 O15 595,16630 449,1086 287,0553 38 * 22,92 Isoquercitrin (Hirsutrin, Quercetin-3-O-glucoside) C21 H20 O12 463,08765 301,0360 300,0282 271,0256 255,0301 39 22,94 3-[(1-Carboxyvinyl)oxy]benzoic acid C10 H8 O5 207,02935 137,0234 135,0442 93,0332 87,0073 40 23,04 Rosmarinic acid-O-hexoside C24 H26 O13 521,12952 359,0778 341,0916 323,0780 197,0452 161,0235 41 23,15 Salvianolic acid isomer C36 H30 O16 717,14557 519,0930 339,0522 321,0408 295,0617 277,0506 42 23,32 Lipedoside A isomer 1 C29 H36 O14 607,20269 461,1674 443,0990 297,0414 161,0235 135,0442 ( Grzegorczyk-Karolak and Kiss, 2018 ) 43 23,58 Lipedoside A isomer 2 C29 H36 O14 607,20269 461,1674 443,0990 297,0430 161,0235 135,0442 ( Grzegorczyk-Karolak and Kiss, 2018 ) 44 23,63 Leucosceptoside A C30 H38 O15 637,21325 461,1673 315,1097 193,0503 175,0393 160,0156 ( Grzegorczyk-Karolak and Kiss, 2018 ) 45 24,02 Apigenin-O-(deoxyhexosyl)hexoside C27 H30 O14 579,17139 433,1135 271,0605 46 * 24,05 Cosmosiin (Apigetrin, Apigenin-7-O-glucoside) C21 H20 O10 433,11347 271,0605 153,0182 119,0494 47 24,08 Apigenin-O-glucuronide C21 H18 O11 445,07709 269,0461 175,0240 113,0232 ( Grzegorczyk-Karolak and Kiss, 2018 ) 48 24,30 Rosmarinic acid (Labiatenic acid) C18 H16 O8 359,07670 197,0452 179,0343 161,0235 135,0441 123,0439 ( Grzegorczyk-Karolak and Kiss, 2018 ) 49 24,32 Methyl ca ff eate C10 H10 O4 195,06574 163,0392 145,0287 135,0445 117,0339 107,0498 50 24,41 Methoxy-trihydroxy fl avone-O-glucuronide C22 H20 O12 477,10331 301,0712 286,0477 258,0528 51 24,76 N-trans-Feruloyltyramine C18 H19 NO 4 314,13924 177,0550 145,0287 121,0652 52 25,84 Martynoside C31 H40 O15 651,22890 475,1838 193,0502 175,0393 160,0157 ( Grzegorczyk-Karolak and Kiss, 2018 ) 53 25,97 Ethyl ca ff eate C11 H12 O4 209,08139 163,0392 145,0286 135,0444 117,0339 107,0858 54 26,20 3-O-Methylrosmarinic acid C19 H18 O8 373,09235 197,0452 179,0344 175,0393 160,0156 135,0441 (continued on next page )
Table 5 (continued ) No. Rt Name Formula [M + H] + [M -H] − Fragment 1 Fragment 2 Fragment 3 Fragment 4 Fragment 5 Literature 55 26,94 Pentahydroxy fl avone C15 H10 O7 301,03483 273,0427 178,9974 151,0030 107,0125 56 * 27,18 Naringenin C15 H12 O5 271,06065 177,0179 165,0179 151,0026 119,0492 107,0128 57 * 27,83 Luteolin (3',4',5,7-Tetrahydroxy fl avone) C15 H10 O6 285,03991 217,0505 199,0399 151,0027 133,0284 107,0127 58 27,87 Di-O-methylellagic acid C16 H10 O8 329,02975 314,0078 298,9843 270,9891 59 28,65 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 1 C17 H14 O6 315,08687 205,0501 163,0393 153,0552 145,0288 135,0446 60 29,24 Luteolin-O-(coumaroyl)hexoside C30 H26 O13 593,12952 285,0412 284,0334 145,0285 61 29,64 Dihydroxycinnamoyl-dihydroxyphenyl-ethenol isomer 2 C17 H14 O6 315,08687 205,0502 163,0392 153,0545 145,0287 135,0445 62 * 29,66 Apigenin (4',5,7-Trihydroxy fl avone) C15 H10 O5 269,04500 227,0353 225,0556 201,0551 151,0028 117,0333 63 29,88 Chrysoeriol (Scoparol, 3'-Methoxy-4',5,7-trihydroxy fl avone) C16 H12 O6 299,05556 284,0333 256,0381 227,0345 151,0024 107,0125 64 30,20 Tri-O-methylellagic acid C17 H12 O8 343,04540 328,0230 312,9998 297,9758 65 30,56 Dimethoxy-trihydroxy(iso) fl avone C17 H14 O7 329,06613 314,1532 271,0254 66 32,49 Methoxy-trihydroxy fl avone C16 H12 O6 299,05556 284,0333 256,0379 133,0284 67 34,38 Dihydroxy-dimethoxy(iso) fl avone C17 H14 O6 313,07122 298,0489 283,0255 255,0302 68 34,49 Genkwanin C16 H12 O5 283,06120 268,0383 239,0354 117,0331 69 35,90 Hydroxy-trimethoxy(iso) fl avone C18 H16 O6 329,10252 314,0788 313,0712 300,0634 285,0761 70 36,56 Hydroxy-tetramethoxy(iso) fl avone C19 H18 O7 359,11308 344,0896 343,0819 329,0660 313,0713 71 36,66 1-Oxomicrostegiol C20 H24 O3 313,18037 295,1697 280,1465 243,1019 ( Rungsimakan and Rowan, 2014 ) 72 36,99 Viroxocin C20 H24 O3 313,18037 295,1697 227,1070 199,0758 ( Rungsimakan and Rowan, 2014 ) 73 38,23 Apigenin-4',7-dimethyl ether (4',7-Dimethoxy-5-hydroxy fl avone) C17 H14 O5 299,09195 284,0684 256,0732 74 38,64 3-Oxomicrostegiol C20 H24 O3 313,18037 295,1697 280,1462 243,1020 ( Rungsimakan and Rowan, 2014 ) 75 40,33 Hexadecanedioic acid C16 H30 O4 285,20659 267,1973 223,2066 76 41,82 Viridoquinone C20 H24 O2 297,18546 279,1748 269,1904 239,1432 ( Rungsimakan and Rowan, 2014 ) * Con fi rmed by standard.
shown in
Fig. 2
A. The antioxidant potential of S. viridis aerial parts has
been reported previously (
Erdemoglu et al., 2006
;
Grzegorczyk-Karolak
and Kiss, 2018
), but this study is, as far as we know, the
first one to
report the antioxidant potent of S. viridis roots extract. Evidence
gath-ered from the literature indicates the presence of diterpenoids in the
roots of S. viridis (
Rungsimakan and Rowan, 2014
). Diterpenoids
iso-lated from an S. barrelieri have been reported to exhibit potent
anti-oxidant activity (
Kolak et al., 2009
). Besides, rosmarinic acid, known
for its potent antioxidant properties (
Adomako-Bonsu et al., 2017
), has
also been identified from S. viridis roots (
Rungsimakan and Rowan,
2014
). The synergistic effect of these phytochemicals along with other
bioactive constituents might justify the observed antioxidant activity.
To control global health problems, enzymes involved are considered
as key targets (
Vujanović et al., 2019
). For instance, the modulation of
α-amylase and α-glucosidase, key enzymes responsible for the digestion
of starch, is considered as an important therapeutic strategy for the
management of postprandial glucose peaks (
Picot and Mahomoodally,
2017
). The need for novel enzyme inhibitors is primarily fueled by the
side e
ffects of currently existing inhibitors. For example, kojic acid, a
potent inhibitor of tyrosinase, is a widely used therapeutic agent for the
management of skin hyperpigmentation conditions. However, the
uti-lization of kojic acid by the pharmaceutical and cosmetic industry to
thwart excessive melanin production has been associated with many
side e
ffects (
Zengin et al., 2015
). Interestingly, phytochemicals have
proved to be natural enzyme inhibitors, thereby opening new avenues
for research and development of novel therapeutic solutions. In this
study, S. viridis extracts obtained by di
fferent extraction techniques
were tested for possible enzyme inhibitory activity against
α-amylase,
α-glucosidase, acetylcholinesterase (AChE), butyrylcholinesterase
(BChE), and tyrosinase. Additionally, this study was aimed at
identi-fying the most suitable extraction technique for the isolation of potent
enzyme inhibitors from S. viridis roots. It has been previously reported
that S. viridis, as part of the herbal mixture, was traditionally used to
manage diabetes type 1 (
Bayat et al., 2017
). Additionally, several Salvia
species, namely S. chloroleuca (
Asghari et al., 2015
), S. mirzayanii
(
Rouzbehan et al., 2017
) and S. sclarea (
Zengin et al., 2018b
) have been
documented to possess
α-amylase and α-glucosidase inhibitory
prop-erties. From data summarized in
Table 7
, S. viridis showed stronger
Table 6
Antioxidant properties of Salvia viridis extracts*.
Extraction method DPPH (mg TE/g) ABTS (mg TE/g) CUPRAC (mg TE/g) FRAP (mg TE/g) Phosphomolybdenum (mmol TE/g) Metal chelating (mg EDTAE/g)
MAE 199.95 ± 4.83c 280.50 ± 6.52b 858.75 ± 7.35c 653.26 ± 13.00c 2.37 ± 0.14b 22.71 ± 1.11a
MAC 218.59 ± 5.47b 293.28 ± 8.78a 922.64 ± 17.21b 705.89 ± 12.12a 2.68 ± 0.07a 15.60 ± 0.57b
SFE 4.89 ± 0.93e 32.59 ± 2.62c 138.61 ± 3.89d 99.94 ± 1.71d 1.34 ± 0.06c 14.19 ± 3.82b
SE 177.46 ± 2.13d 276.35 ± 6.83b 910.06 ± 7.39b 674.37 ± 5.48b 2.40 ± 0.04b 20.33 ± 1.17a
UAE 240.00 ± 4.57a 302.85 ± 7.12a 970.74 ± 6.58a 704.27 ± 8.09a 2.84 ± 0.01a 19.53 ± 0.90a
* Values expressed are means ± S.D. of three parallel measurements. MAE: Microwave assisted extraction; MAC: Maceration, SFE: Supercriticalfluid extraction,
SE: Soxhlet extraction; UAE: Ultrasonic assisted extraction. TE: Trolox equivalent; EDTAE: EDTA equivalent. Different letters indicate differences in the tested extracts (p < 0.05).
Fig. 2. Statistical evaluations (A: Correlation coefficients between total bioactive compounds and biological activities (Pearson Correlation Coefficient (R), p < 0.05); B: Distribution of biological activities on the correlation circle based on PCA; C: Heatmap of extracts in according to bioactive compounds and biological activities; D: Distribution of the extraction methods on the correlation circle based on PCA, individual sample replications (n = 4) are given in the class prediction
inhibitory action against
α-glucosidase compared to α-amylase. It has
been proposed that low
α-amylase versus strong α-glucosidase
inhibi-tion could address the major drawbacks of currently used hypoglycemic
agents. Indeed, excessive inhibition of
α-amylase has been found to
result in bacterial fermentation of undigested carbohydrate in the
colon, eventually causing gastrointestinal problems (
Picot and
Mahomoodally, 2017
). The root extracts of S. viridis showed inhibitory
activity against both AChE and BChE (
Table 7
). The observed inhibitory
activity against AChE might be related to the presence of
β-sitosterol
(
Bahadori et al., 2016
) which has been previously identified from S.
viridis roots (
Rungsimakan and Rowan, 2014
). It is noteworthy
men-tioning that S. viridis ethanolic root extract obtained by SFE, showed
higher inhibitory action against both cholinesterases (5.21 and 6.33 mg
GALAE/g). A group of authors (
Lee et al., 2011
) previously reported the
tyrosinase inhibitory activity of S. viridis aerial parts. Data gathered
from this study demonstrated that S. viridis roots also exhibited
tyr-osinase inhibitory activity (
Table 7
). Tyrosinase, responsible for
mela-nogenesis, is the main target for the management of epidermal
hy-perpigmentation problems (
Zengin et al., 2018c
). Other species of the
Salvia genus have also been reported to possess tyrosinase inhibitory
activity. For instance, S. sclarea water extract, ethanolic extract of S.
cryptantha and S. cyanescens aerial parts (
Süntar et al., 2011
;
Zengin
et al., 2018b
). Furthermore, rosmarinic acid, present in several species
of the Salvia genus, identi
fied in S. viridis roots has been reported to
exhibit inhibitory action against tyrosinase (
Oliveira et al., 2013
).
To provide new insights into the extraction methods in this study
performed, we used several statistical analysis. In Principal Component
Analysis (PCA), two principal components were obtained as 89.5% of
total variance (
Fig. 2
B). The PCA and correlation analysis were similar
and strong correlation was observed between total bioactive
compo-nents and DPPH, ABTS, phosphomolybdenum, CUPRAC and FRAP
as-says. To provide a certain classification for the extraction methods, a
heat map (
Fig. 2
C) was carried out, as well as PCA (
Fig. 2
D). Based on
these results, the extraction methods were divided into three groups.
SE, MAC, and UAE were classified in the same group. However, other
groups contained SFE and MAE.
4. Conclusion
This study has revealed the antioxidant and enzyme inhibitory
properties of S. viridis ethanolic root extracts, for the
first time. It was
noted that the choice of extraction technique in
fluenced the antioxidant
properties of S. viridis ethanolic root extracts. This study appraises the
possible therapeutic application of S. viridis ethanolic root extracts for
the management of chronic complications such as Alzheimer’s disease,
diabetes, and skin hyperpigmentation disorders. Additionally, scienti
fic
evidence gathered in this investigation support the need for a further
study which might lead to the development of new therapeutic entities
for the management of the ailments as mentioned above.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:
https://doi.org/10.1016/j.indcrop.2019.01.060
.
References
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Table 7
Enzyme inhibitory properties of Salvia viridis extracts*.
Extraction method AChE inhibition (mg GALAE/g)
BChE inhibition (mg GALAE/g)
Tyrosinase inhibition (mg KAE/g)
α-Amylase inhibition (mmol ACAE/g)
α-Glucosidase inhibition (mmol ACAE/g) MAE 4.08 ± 0.05b 5.63 ± 0.02b 159.75 ± 1.80a 0.56 ± 0.03c 1.66 ± 0.01a MAC 3.60 ± 0.15c 5.45 ± 0.05c 162.56 ± 0.78a 0.73 ± 0.01a 1.63 ± 0.01b SFE 5.21 ± 0.01a 6.33 ± 0.02a 151.54 ± 2.14c 0.73 ± 0.01a 1.64 ± 0.01ab SE 4.23 ± 0.07b 5.60 ± 0.04b 159.58 ± 0.35a 0.66 ± 0.02b 1.61 ± 0.02b UAE 3.62 ± 0.20c 5.44 ± 0.01c 158.38 ± 0.14b 0.72 ± 0.03a 1.65 ± 0.01a
* Values expressed are means ± S.D. of three parallel measurements. GALAE: Galatamine equivalent; KAE: Kojic acid equivalent; ACAE: Acarbose equivalent.
MAE: Microwave assisted extraction; MAC: Maceration, SFE: Supercriticalfluid extraction, SE: Soxhlet extraction; UAE: Ultrasonic assisted extraction. TE: Trolox
