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Contents of phenolic and flavonoid compounds

in Isatis demiriziana Mısırdalı: an endemic to
the Southeast Anatolia, Turkey

Article · April 2017

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 www.biodicon.com Biological Diversity and Conservation

ISSN 1308-8084 Online; ISSN 1308-5301 Print 10/1 (2017) 115-123

Research article/Araştırma makalesi

Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast
Anatolia, Turkey

Özgür KARAKAŞ *1, Mustafa Abdullah YILMAZ 2, Hilal SURMUŞ ASAN 3, Ahmet ONAY 3, Nazan ÇALAR 3
1
Şırnak University, Faculty of Agriculture, Department of Agricultural Biotechnology, 73300 Şırnak, Turkey
2
Dicle University Science and Technology Research and Application Center (DÜBTAM), Dicle University, 21280
Diyarbakır, Turkey
3
Department of Biology, Faculty of Science Dicle University, 21280 Diyarbakir, Turkey

Abstract
Isatis demiriziana Mısırdalı plant contains a number of compounds which has anticarcinogenic, antioxidant
and other preventive effects. In this study, the flavonoid and phenolic contents in the plant samples harvested in the
vegetative (leaf and root) and full flowering season (flower, leaf and root) were determined by LC-MS/MS. Among the
27 compounds studied, malic acid was found to be the most abundant compound in the methanolic extracts of samples
and the amount of malic acid of vegetative root extracts were the highest (30124,37 µg g -1 dry-extract). Moreover, it
was also determined considerable amount of salicylic acids and p-coumaric in the root extracts. This study is the first
detailed study on the phenolic and flavonoid compounds of I. demiriziana. Based on the findings of this study, in
further researches might be refered as an additional source in production of phenolic and flavonoid compounds.

Key words: Isatis, LC–MS/MS, phenolics, flavonoids, endemic

----------  ----------

Türkiye’de Güneydoğu Anadolu Bölgesi’ne endemik Isatis demiriziana Mısırdalı’ndaki fenolik ve flavonoid
bileşiklerinin içerikleri

Özet
Isatis demiriziana Mısırdalı bitkisi antikanserojen, antioksidan ve başka koruyucu özelliklere sahip olan çok
sayıda bileşik içermektedir. Bu çalışmada, I. demiriziana’nın vejetatif (yaprak ve kök) ve tam çiçeklenme (çiçek, yaprak
ve kök) dönemlerinde hasat edilen bitki örneklerinin flavonoid ve fenolik içerikleri LC-MS/MS ile tespit edildi.
Çalışılan 27 bileşik arasında, örneklerin metanolik ekstraktlarında malik asit miktarı en fazla düzeyde bulundu ve
vejetatif köklerdeki malik asit miktarı (30124,37 µg g-1 kuru ekstrakt) en yüksek orana sahipti. Ayrıca kök
ekstraktlarında büyük miktarda salisilik asit ve p-kumarik asit tespit edildi. Bu çalışma, I. demiriziana’nın fenolik ve
flavonoid içerikleri üzerine yapılmış ilk detaylı çalışmadır. Bu çalışmanın sonuçlarına dayanılarak, ileriki çalışmalarda
fenolik ve flavonoid bileşiklerin üretiminde bir ek kaynak olarak başvurulabilir.

Anahtar Kelimeler: Isatis, LC–MS/MS, fenolikler, flavonoidler, endemik

1. Introduction

Brassicaceae (Cruciferea) has about 350 genera and 3000 species, growing mostly in the North Temperate
Zone and Mediterranean Region (Mabberley, 1987). Isatis genus belongs to Brassicaceae family and this genus is
represented by 40 taxa which 24 of these are endemic to Turkey (Davis, 1988). Moreover, this genus has 31 species and
14 subspecies in Eastern and South-Eastern Anatolia (Mısırdalı, 1985). The chemical constitutients extracted from the
roots and leaves of Isatis species have antiviral, anticancer, antibacterial, astringent, febrifuge and antirheumatic effects
(Radwan et al., 2008; Vang, 1994; Kirtikau and Basu, 1983; Bown, 1995). They are also employed for different

*
Corresponding author / Haberleşmeden sorumlu yazar: Tel.: +904865513032; Fax.: +904865513031; E-mail: [email protected]
© 2008 All rights reserved / Tüm hakları saklıdır BioDiCon. 612-1116
116 Biological Diversity and Conservation – 10 / 1 (2017)

disorders such as encephalitis, meningitis, erysipelas, influenza, heat rash, mumps in the traditional medicine. It is
reported that uses of Isatis species ethnobotanical in Eastern Anatolia (Polat et. al., 2012). Use of natural antioxidants
including phenolic acids and flavonoids as preventive and therapeutic drug have been attracting considerable attention
because of their antioxidant properties and anticarcinogenic potential. These compounds have several health promoting
influences (Costa et al., 2012; Erdogan-Orhan et al., 2014; Prakash et al., 2007). Phenolic compounds act as
antioxidants (Maillard et al., 1996). Phenolic compounds play a significant role in total antioxidant capacity of
vegetables, fruits and grains (Jacobo-Velazquez and Cisneros-Zevallos, 2009). The benefits of flavonoids including
reducing the risk of atherosclerosis developing have been clearly shown (Knekt et al., 1996). Flavonoids perform some
important functions such as cell cycle inhibition, nitrogen fixation and filtration of UV rays (Kumar and Pandey, 2013).
Several studies suggested that flavonoids have protective impacts against degenerative diseases such as cardiovascular
diseases, cancers and other age-related diseases (Pandey, 2007; Kumar et al., 2013). Phenolic acids include two primary
groups: Cinnamic and benzoic acid (Tarnawski et. al., 2006). The chlorogenic acid is the most significant cinnamic acid
which is a combination of quinic and caffeic acids. Hydroxybenzoic acid derivatives are firstly present as vanillic acids
but p-hydroxybenzoic and protocatechuic acids are more extensive (Kvasnicka et. al., 2008).
Study on several certain phenolic acids such as gallic acid, malic acid, p-coumaric acid, vanillic acid and
syringic acid reveals that they have numerous advantage for human health. Malic acid plays a vital role in reversing
muscle fatigue and mental clarity. Moreover, these actions can make it a beneficial treatment for sufferers of
fibromyalgia (Russell et al., 1995). Vanillic acid and p-coumaric acid have hydroxide scavenging activity (Kang et al.,
2006). Moreover, p-coumaric acid is a potent inhibitor of 5-S-cysteinyl dopamine induced neurotoxicity and this
compound is used in treatment of Parkinson’s disease (Vauzour et al., 2010). It was shown that the Gallic acid acted as
an important agent in protection of renal damage causing death of tumor cells (Canbek et al., 2013). Salicylic acid plays
a significant role in conservation against virus infection by inhibiting catalase resulting in the accumulation of H2O2 in
plant cells (Chen et al., 1993).
Different parts of plants vary in terms of contents of phenolics and flavonoids, so each plant part needs to be
analyzed to identify its potential advantage as a health product. It was reported that the chemical constituents in the
leaves of some Isatis species possess antibacterial, antiviral, anticancer, febrifuge and astringent features (Karakoca et
al., 2013) but there is no report on the phenolic and flavonoid contents of I. demiriziana. In this research, we aimed to
determine the phenolic and flavonoid contents of different parts of I. demiriziana plants collected in the vegetative
season (leaf and root) and full flowering season (flower, leaf and root). Liquid chromatography–tandem mass
spectrometry (LC– MS/MS) technique was used to analyze the phenolic and flavonoid contents of I. demiriziana.

2. Materials and methods

2.1. Plant Material

Five different samples for I. demiriziana were collected from a height of 1300 meters from Ergani county of
the Diyarbakır province and at vegetative (leaf and root) and full flowering season (flower, leaf and root). Voucher
specimens were deposited at the Herbarium of Dicle University, Faculty of Science (voucher no. DUF-6050).
Specimens were identified by Prof. Dr. Ömer SAYA, from the same institution. The samples were air-dried at root
temperature and then the samples were pulverized with a laboratory mill and stored at 40C until the chemical tests were
conducted. Quantitation and identification of phenolic and flavonoid compounds

2.2.1. Plant extract preparation for LC–MS/MS

The collected specimens were dried by air at room temperature, which were then powdered. The samples (100
g) were extracted three times with 300 mL of methanol for 24 h. Then, a rotary evaporator was used for removal of the
solvent at 30◦C. The remaining solid (Yield: 15.6%) was used to prepare a 1000 mg L-1 solution, which was then
filtrated with a 0.2 µm microfiber filter to use for LC–MS/MS analysis.

2.2.2. Instruments and chromatographic conditions for LC–MS/MS

The phenolics were analyzed quantitatively by LC-MS/MS (Shimadzu LC/MS 8040 model). The liquid
chromatograph has DGU-20A3R degasser, LC-30AD binary pumps, SIL-30AC autosampler and CTO-10ASvp column
oven. The samples were separated chromatographically on a C18 reversed-phase Inertsil ODS-4 (150 mm × 4.6 mm, 3
µm) analytical column (40◦C). The elution gradient comprised of mobile phase A:water, 0.1% formic acid and 5 mM
ammonium formate and mobile phase B: methanol, 0.1% formic acid and 5 mM ammonium formate. The gradient
program with the following ratios of solvent B was carried out t (min), B%: (0, 40), (20, 90), (23.99, 90), (24, 40), (29,
40). The solvent flow percentage was continued at 0.5 mL/min and injection volume was adjusted as 4 µL.

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey
 Biological Diversity and Conservation – 10 / 1 (2017) 117

2.2.3. MS instrumentation

The samples were analyzed by MS employing a Shimadzu LC/MS 8040 model triple quadrupole mass
spectrometer equipped (ESI source operating in both negative and positive ionization modes). Data obtained from LC–
MS/MS were evaluated by Lab Solutions software (Shimadzu, Kyoto, Japan). The analyses were quantified by the
multiple reaction monitoring (MRM) mode: the studied compounds were assayed following two or three transitions for
each compound, the first one for quantitative uses and the second and/or the third one for checking of the finding. The
optimum ESI conditions were determined as interface temperature; 350◦C, DL temperature; 250◦C, heat block
temperature; 400◦C, nebulizing gas flow (nitrogen); 3 L/min and drying gas flow (nitrogen); 15 L/min.

2.2.4. Analytical parameters for the validated LC–MS/MS method

Table 1 shows rectilinear regression equations and the linearity ranges of the studied standard chemicals.
Correlation coefficients were higher than 0.99. Table 1 also displays the limit of detection (LOD) and limit of
quantitation (LOQ) of the analytical method. LOD values of the compounds are between 0.05 and 25.8 g/L and LOQ
values are between 0.17 and 85.9 g/L (the recoveries of the phenolics are between 96.9% and 106.2%).

Table 1. Analytical parameters of UHPLC-ESI-MS/MS method

Parent Ioniza Linearity

RSD% LOD/LOQ Recove
No Analytes RTa ion tion R2c Range Uf
d (µg/L)e ry (%)
(m/z)b Mode (mg/L)

1 Quinic acid 3.32 190.95 Neg 0.9927 0.0388 250-10000 22.3 / 74.5 103.3 4.8
2 Malic acid 3.54 133.05 Neg 0.9975 0.1214 250-10000 19.2 / 64.1 101.4 5.3
3 tr-Aconitic acid 4.13 172.85 Neg 0.9933 0.3908 250-10000 15.6 / 51.9 102.8 4.9
4 Gallic acid 4.29 169.05 Neg 0.9901 0.4734 25-1000 4.8 / 15.9 102.3 5.1
5 Chlorogenic acid 5.43 353 Neg 0.9932 0.1882 250-10000 7.3 / 24.3 99.7 4.9
6 Protocatechuic acid 5.63 152.95 Neg 0.9991 0.5958 100-4000 25.8 / 85.9 100.2 5.1
7 Tannic acid 6.46 182.95 Neg 0.9955 0.9075 100-4000 10.2 / 34.2 97.8 5.1
8 tr- caffeic acid 7.37 178.95 Neg 0.9942 1.0080 25-1000 4.4 / 14.7 98.6 5.2
9 Vanillin 8.77 151.05 Neg 0.9995 0.4094 250-10000 10.1 / 33.7 99.2 4.9
10 p-Coumaric acid 9.53 162.95 Neg 0.9909 1.1358 100-4000 15.2 / 50.8 98.4 5.1
11 Rosmarinic acid 9.57 358.9 Neg 0.9992 0.5220 250-10000 10.4 / 34.8 101.7 4.9
12 Rutin 10.18 609.1 Neg 0.9971 0.8146 250-10000 17.0 / 56.6 102.2 5.0
13 Hesperidin 9.69 611.1 Poz 0.9973 0.1363 250-10000 21.6 / 71.9 100.2 4.9
14 Hyperoside 10.43 463.1 Neg 0.9549 0.2135 100-4000 12.4 / 41.4 98.5 4.9
15 4-OH Benzoic acid 11.72 136.95 Neg 0.9925 1.4013 25-1000 3.0 / 10.0 106.2 5.2
16 Salicylic acid 11.72 136.95 Neg 0.9904 0.6619 25-1000 4 / 13.3 106.2 5.0
17 Myricetin 11.94 317 Neg 0.9991 2.8247 100-4000 9.9 / 32.9 106.0 5.9
18 Fisetin 12.61 284.95 Neg 0.9988 2.4262 100-4000 10.7 / 35.6 96.9 5.5
19 Coumarin 12.52 146.95 Poz 0.9924 0.4203 100-4000 9.1 / 30.4 104.4 4.9
20 Quercetin 14.48 300.9 Neg 0.9995 4.3149 25-1000 2.0 / 6.8 98.9 7.1
21 Naringenin 14.66 270.95 Neg 0.9956 2.0200 25-1000 2.6 / 8.8 97.0 5.5
22 Hesperetin 15.29 300.95 Neg 0.9961 1.0164 25-1000 3.3/ 11.0 102.4 5.3
23 Luteolin 15.43 284.95 Neg 0.9992 3.9487 25-1000 5.8 / 19.4 105.4 6.9
24 Kaempferol 15.43 284.95 Neg 0.9917 0.5885 25-1000 2.0 / 6.6 99.1 5.2
25 Apigenin 17.31 268.95 Neg 0.9954 0.6782 25-1000 0.1 / 0.3 98.9 5.3
26 Rhamnetin 18.94 314.95 Neg 0.9994 2.5678 25-1000 0.2 / 0.7 100.8 6.1
27 Chrysin 21.18 253 Neg 0.9965 1.5530 25-1000 0.05 / 0.17 102.2 5.3

RT: Retention time

b
Parent ion (m/z): Molecular ions of the standard compounds (mass to charge ratio)
c 2
R : coefficient of determination
d
RSD: relative standard deviation
e
LOD/LOQ (µg/L): Limit of detection/Limit of quantification
f
U (%): Percent relative uncertainty at 95% confidence level (k=2).
g
Values in µg g-1 (w/w) of plant methanol extract
h
N.D: not detected.

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey
118 Biological Diversity and Conservation – 10 / 1 (2017)

Figure 1. LC-MS/MS chromatograms of 250 ppb standard mix. quinic acid: 1, malic acid: 2, tr-aconitic acid:3, gallic
acid:4, chlorogenic acid:5, protocatechuic acid:6, tannic acid:7, tr-caffeicacid:8, vanillin:9, p-coumaric acid:10,
rosmarinic acid:11, rutin:12, hesperidin:13, hyperoside:14, 4-OH benzoic acid:15, salicylic acid:16, myricetin:17,
fisetin:18, coumarin:19, quercetin:20, naringenin:21, hesperetin:22, luteolin:23, kaempferol:24, apigenin:25,
rhamnetin:26, chrysin:27.

Figure 2. LC-MS/MS chromatogram of leaf in vegetative season.

Figure 3. LC-MS/MS chromatogram of root in vegetative season

Figure 4. LC-MS/MS chromatogram of root in full flowering season

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey
 Biological Diversity and Conservation – 10 / 1 (2017) 119

Figure 5. LC-MS/MS chromatogram of flower in full flowering season

Figure 6. LC-MS/MS chromatogram of leaf in full flowering season

Figure 7. LC-MS/MS chromatogram of fruit

3. Results

3.1. Quantitative analysis of phenolics, flavonoids compounds by UHPLC-ESI-MS/MS

Results of phenolic and flavonoid content in the five samples of I. demiriziana have been presented in Table 2.
Significant differences in the phenolic and flavonoid constituents of the different extracts of I. demiriziana were
observed. LC–MS/MS analysis obviously display that the methanol extracts of I. demiriziana contain many phenolic
and non-phenolic compounds Figure 2-7. Phenolic acids exist in most plants, and each plant can be adequately specific
for the availability of various phenolic acids and their derivatives together with the other groups (Ziakova et. al., 2003).
It was determined that the main components of all plant samples were malic acid, quinic acid, tr-aconitic acid,
vanillin, p-coumaric acid, 4-OH benzoic acid, salicylic acid, protocatechuic acid, tannic acid and tr-caffeic acid
compounds. Out of non-phenolic compounds, I. demiriziana extracts include high amounts of quinic acid (221.18-
1538.49 µg g-1), tr-Aconitic acid (105.2 -298.62 µg g-1), salicylic acid (35.94-216.27 µg g-1), p-coumaric acid(19.13-
1457.45 µg g-1) and 4-OH Benzoic acid (27.99-176.57µg g-1) and lower amounts of gallic acid (0.58-2.05 µg g-1) (Table
2). Malic acid (MA) was single dominant compound among all samples studied. Among them, the vegetative root gave

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey
120 Biological Diversity and Conservation – 10 / 1 (2017)

the highest amount of malic acid with a 30124.37 µg g-1 extract (Table 2, Figure 3). This was followed by the flowering
root, flower, the flowering leaf, the vegetative leaf and fruit stage with the amounts of 27733.72, 14.438.42, 6879.07,
3745.49 and 691.3 µg g-1 extract, respectively. Malic acid contents of the five samples of I. demiriziana were so
different. This variation may be due to different organ or different growing stages of the samples studied and to gain
more insight how growing stages and different organs influence phenolic content of this plant more studies are required.
In prompting plant defense responses, present report declares that the metabolic levels of MA compounds play an
important role (Huckelhoven, 2007). A corresponding induced defense response beginning intraplant signaling between
roots and leaves was implicated in herbivory (Rasmann et al., 2005).
Quinic acid is a metabolite that responsible for metabolic response (inducible defense) to biotic stress (Murthy
et al., 2009). It was the second highest phenolic acid determined as 994.56, 550.74, 1087.19, 221.18, 1538.49 and
487.62 µg g-1 extract in the vegetative leaf, vegetative root, flowering leaf, flowering root, flower and fruit samples,
respectively. Besides, the amount of quinic acid that obtained from leaf extracts (vegetative leaf 994.56 µg g-1 and
flowering leaf 1087.19 µg g-1) were greater amounts from root extracts (vegetative root 550.74µg g-1 and flowering
root 221.18 µg g-1). Similiarly, it is reported that the quinic acid and quercitol are present in high concentrations in
wounded leaves of genus Quercus plants (Gargallo-Garriga et al., 2010). It is known that the trans-aconitic acid has
antirheumatic and diuretic properties (Schnitzler, 2007) althought the distribition of this compound is rare (Nierhaus
and Kinzel, 1971). The highest amount of trans-aconitic acid was obtained from flower stage with 298.62 µg g-1 (Figure
5). Salicylic acid (SA) is believed to be a plant signal molecule playing an important role in plant, development, growth
and defense responses, and functioning in the commencement of systemic attained resistance (SAR) (Hahlbrock and
Scheel, 1989; Ding et al., 2002). The vegetative and flowering root extracts contained significant amount of vanillin
(124.09 and 311.94 µg g-1), protocatechuic acid (59.4 - 66.91 µg g-1) , tannic acid (40. 98- 25. 92 µg g-1) and tr-caffeic
acid (34. 09- 29. 41µg g-1) respectively (Table 2, Figure 3,4). Vanillin is the main constituent of natural vanilla, a well-
known food and cosmetic additive and has antioxidant and antimutagenic properties (Davidson and Naidu, 2000).
Their collection is extremely sensitive to environmental situations such as water, light and nutrient availability, and
pathogen infection (Harvell and Bosland, 1997). PCA is a natural phenolic acid and exist in several plants including
mushrooms and microorganisms (Williams et al., 2012; Nguyen et al., 2013). It is known that the PCA has
antiinflammatory and antioxidant (Liu et al., 2002; Syafni et al., 2012) and antibiotic activities (Nguyen et al., 2015).
Tannic acid has antioxidant (Andrade et al., 2005), antimutagenic (Ferguson, 2001) and anticarcinogenic properties
(Nepka et al., 1999). It is reported that the tannic acid induced by Rhizobia in rice, which is resistant to Rhizoctonia
(Mishra et al., 2006).
Phenolic acid and flavonoids in plants have various functions such as protein synthesis, nutrient uptake,
photosynthesis, allelopathy, enzyme activity, and structural components (Hung, 2016). Flavonoids are the largest group
of phenolics having antimicrobial and antioxidant impacts (Lorenc et al., 2005). Along with their roles in plants, these
compounds in human diet might introduce a number of benefits connected with reduced risk of chronic diseases
including anti-inflammatory, anti-atherogenic, antiallergenic, antioxidant, anti-thrombotic, anti-microbial, vasodilatory
and cardioprotective influences (Manach et al., 2004). In a study, Nahak et al. (2014) indicated that the phenolic
constituent of a plant is usually a good sign of its antioxidant potential. It is found that the flower extracts of I.
demiriziana have highest levels of flavonoids quercetin (7.98 µg g-1), naringenin (22.96 µg g-1), rhamnetin (72.74 µg g-
1
), and hyperoside (49.19 µg g-1) and non phenolics tr-aconitic acid (298.62 µg g-1), and p-coumaric acid (1457.45 µg g-
1
). Recently, Chang et al., (2016) stated that the acid hydrolysis extract of I. indigotica contained 61.02 mg/100g of p-
coumaric acid, and 23.13 mg/100g of gallic acid. Similar propensity was also viewed for the flavonols, quercetin and
hyperoside. The rutin mostly gathered at the fruiting and flowering phases (9.25- 29.67 µg g-1 respectively). Because
they move as attractive to pollinators and/or to protect the reproductive structures against UV radiations and herbivores,
probably, this accumulation pattern of quercetin, hyperoside and rutin may be associated with their biological roles
(Gronquist et al., 2001; Kreft et al., 2003). Kaempferol (1.13- 0.13 µg g-1), hesperetin (0-0.21 µg g-1), luteolin (0-0.42
µg g-1) and apigenin(0-0.35 µg g-1) were the most abundant flavonoids in the exracts of I. demiriziana (Table 2).
According to our result, the highest amounts of hesperidin (27.61 µg g-1), chlorogenic acid (110.4 µg g-1), rutin
(29.67 µg g-1), 4-OH benzoic acid (176.57 µg g-1) and salicylic acid (216. 27 µg g-1) were obtained from the fruit stage
(Table 2, Figure 7). Hesperidin (Hsd) and hesperetin (Hst) have several biological activity such as antioxidant, anti-
inflammatory and anti-cancer impacts. To struggle with different pathogens, these compounds play an important role in
plant defense systems (Soares et al., 2015). Chlorogenic acid is widely employed in industries and medicine including
food industries and the consumer chemicals (Kweon et al., 2001). It is a natural antioxidant and anticancer agent and
has antiviral and antibacterial properties (Jiang et al., 2001). Karakoca et al., (2013) determined that the chlorogenic
acid content was 1980.20 µg g-1 on the methanolic root extract of I. floribunda. Among twenty-seven references used;
rosmarinic acid, myricetin, coumarin, fisetin and chrysin were not detected in I. demiriziana extracts employed in this
study.

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey
 Biological Diversity and Conservation – 10 / 1 (2017) 121

Table 2. Quantitative analysis of phenolic and flavonoid by LC-MS/MS in I. demiriziana (µg analyte/g extract)
Compounds Vegetative-leaf Vegetative-root Flowering-leaf Flowering-root Flower Fruit
Hesperidin 1.65 0.71 8.72 0.6 9.73 27.61
Coumarin 0 0 0 0 0 0
Quinic acid 994.56 550.74 1087.19 221.18 1538.49 487.62
Malic acid 3745.49 30124.37 6879.07 27733.72 14438.42 691.3
tr-Aconitic acid 105.15 129.5 187.85 114.53 298.62 105.2
Gallic acid 1.85 1.92 1.72 1.79 2.05 0.58
Chlorogenic acid 0.58 0.3 10.59 0.4 0.24 110.4
Protocatechuic acid 21 59.4 35.12 66.91 21.53 11.08
Tannic acid 4.53 40.98 25.68 25.92 4.53 4.63
tr-caffeic acid 2.35 34.09 2.83 29.41 7.08 1.86
Vanillin 26.31 124.09 24.64 311.94 6.39 133.67
Rosmarinic acid 0 0 0 0 0 0
p-Coumaric acid 77.77 129.35 98.44 85.92 1457.45 19.13
Rutin 0.46 0.69 5.41 0.20 9.25 29.67
Hyperoside 25.54 0.17 30.02 0.51 49.19 3.79
Myricetin 0 0 0 0 0 0
Fisetin 0 0 0 0 0 0
4-OH Benzoic acid 62.68 132.03 96.75 87.03 27.99 176.57
Salicylic acid 77.24 155.74 111.49 102.02 35.94 216.27
Quercetin 2.12 0.15 5.93 0 7.98 0.33
Kaempferol 1.13 0 0.13 0 0 0
Naringenin 1.26 0.04 1.5 0.04 22.96 0.16
Hesperetin 0.06 0.08 0.14 0 0.21 0.04
Luteolin 0.42 0 0.37 0.34 0 0.32
Apigenin 1.04 0.17 0.35 0.02 0 0.09
Rhamnetin 2.13 0 7.44 0 72.74 3.53
Chrysin 0 0 0 0 0 0

4. Conclusions and discussion

The present study can be deduced as the phenolic contents of different organs of I. demiriziana compared
favorably with other plants. According to the results of this study, further investigations on I. demiriziana may be
carried out to identify factors that will affect phenolic and flavonoids level in plant tissues, thus they might be grown
and harvested under optimum circumstances to increase pleasing qualities chemical levels.

Acknowledgements

The authors would like to thank Dicle University research fund (DUBAP, project no:06-FF-141).

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(Received for publication 21 October 2016; The date of publication 15 April 2017)

Özgür KARAKAŞ et al., Contents of phenolic and flavonoid compounds in Isatis demiriziana Mısırdalı: an endemic to the Southeast Anatolia, Turkey

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