The Scientific World JournalVolume 2012 (2012), Article ID 564367, 12 pagesdoi:10.1100/2012/564367

Research Article

Phenolic Compounds Analysis of Root, Stalk, and Leaves of Nettle

Semih Otles and Buket Yalcin

Food Engineering Department, Engineering Faculty, Ege University, 35100 Bornova/Izmir, Turkey

Received 17 October 2011; Accepted 19 December 2011

Academic Editor: Yiannis Fiamegos

Copyright © 2012 Semih Otles and Buket Yalcin. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract.

Types of nettles (Urtica dioica) were collected from different regions to analyze phenolic compounds in this research. Nettles are specially grown in the coastal part. According to this kind of properties, nettle samples were collected from coastal part of (Mediterranean, Aegean, Black sea, and Marmara) Turkey. Phenolic profile, total phenol compounds, and antioxidant activities of nettle samples were analyzed. Nettles were separated to the part of root, stalk, and leaves. Then, these parts of nettle were analyzed to understand the difference of phenolic compounds and amount of them. Nettle (root, stalk and leaves) samples were analyzed by using High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) to qualitative and quantitative determination of the phenolic compounds. Total phenolic components were measured by using Folin-Ciocalteu method. The antioxidant activity was measured by using DPPH (2,2-diphenyl-1-picrylhydrazyl) which is generally used for herbal samples and based on single electron transfer (SET).

1. Introduction

Nettle (Urtica dioica L.), naturally found in pathway, field, and wildwood. Nettles are grown in mild climate areas, bottom of barriers, ruins and grassy places, between cultivated plants, street, and water runnels. This plant prefers nutrient riches and lighted places, hot and mild climate. It has broad distribution of the world. It is also grown in the different region, of Turkey. Some local names of nettle are “dalagan, dizlagan, agdalak, and isirgi”. Nettle is annual or perennial, herbaceous plant. This plant which is specially grown in Black Sea region has reach chemical composition. It is used as drug, food, fibrous, dye, and cosmetic from centuries. Numbers of medical and pharmacologic researches about nettle are increased day by day. On the other hand, nettle has valuable fibrous content which is light, elegant, long, and resistant. In Turkey, nettle could be an alternative product in Black Sea region in which it could be grown easily [1].

Nettle has dark green leaves, root, stem, serration, and stinging nettle. The nettle flowers are small and green. It could give flowers from May to September. The fruits of nettle are arid and single germ. It has two species. Both of them have 2–4 cm long, oval, and core shape leaves. Fresh nettle could cause blushing and burning of skin when it is touched [2].

Plants could be used as a cure for different types of diseases for centuries. In recent years, usage of plant is increased. In Turkey, plants could be used as household and herbal remedy. Phenolic compounds could be defined as biologically active and herbal and have positive effects on health. The scientific researches are increased about the positive effect of phenolic compounds into coronary heart disease and high blood pressure, diabetes, cancer, inflammative, viral and parasitic disease, psychotic disorders [3].

Systematic properties of nettle could be classified as [4]; Scientific name: Urtica dioica. Traditional name: Nettle.Genus: Urticaceae.Used parts: root, stalk, and leaves.Usage as a food: tea (root, stalk, and leaves), vegetable dish (stalk and leaves), salad (stalk and leaves), and so forth.

A research about nettle indicates that it has wide field of usage as household remedy in Italy from gastrointestinal diseases to rheumatism pains [5]. Another research indicates household remedy usage in Moorish, stalk, and leaves of nettle used in treatment of diabetes, hypertension, astringent, antirheumatic, diuretic, antidiuretic, and cholagogue [6].

Nettle has agglutinin, acetophenone, alkaloids, acetylcholine, chlorogenic acid, butyric acid, chlorophylll, caffeic acid, carbonic acid, choline, histamine, coumaric acid, formic acid, pantothenic acid, kaempferol, coproporphyrin, lectin, lecithin, lignan, linoleic and linolenic acids, palmitic acid, xanthophyll, quercetin, quinic acid, serotonin, stigmasterol, terpenes, violaxanthin, and succinic acid in its chemical content. Nettle also contains 2,5% fatty substance, 14–17% albumins, and 18% protein in dry matter. Seeds of nettle contain 8–10% fixed oil. 1 kg fresh plant contains 130 mg vitamin C, 730 mg carotene, and oxalate. Stinging hair of nettle contains formic acid, histamine, and acetylcholine. Leaves of nettle contain provitamin A, vitamin B₁, K, xanthophylls, and sistosterin and ashes of nettle contain 6,3% ferric oxide, potassium, calcium, and silicium [1].

Analysis of methanolic extracts of nettle was made by using Gas Chromatography-Mass Spectrometry (GC-MS) method; vanillic acid, homovanillic acid, 2-hydroxycinnamic acid, 4-hydroxycinnamic acid, and ferulic acid were found [7]. Methanolic extract of nettle leaves analysis was made by using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method, UV detector; syringic acid, gallic acid, and ferulic acid were found [8].

In a research about nettle root, analysis was done by using extract which were prepared in different pH of organic solvent and by using GC-MS method. The results indicate that roots have 18 different phenolic components and 8 different lignan components. But the chemical composition of these components was not identified in this study [9]. In another research, some kind of components (ferulic acid 20 μg/g, homovanillly alcohol 8 μg/g, and p-coumaric acid 5 μg/g) were identified by using isocratically fractionate, commercial Urtica dioica root extracts without hydrolisation [10]. In another research, 7 flavonoid glycosides (kaempferol-3-O-glycoside, quercetin-3-O-glycoside, isorhamnetin-3-O glycoside, quercetin-3-O-rutinoside, isorhamnetin-3-O-rutinoside, kaempferol-3-O-rutinoside and isorhamnetin-3-O-neohesperidoside) were isolated from Urtica dioica flowers. Structure of these components was identified by using chromatographic and spectroscopic method [11].

Methanolic extracts of nettle leaves and stalks were studied about anthocyanin glycosides, and 3 different components (pelargonidin xylobioside, pelargonidin monoxyloside, and another component which give pelargonidin, D-glucose, and L-rhamnose after acid hydrolysis) were isolated [12]. In addition to the 7 different components, quercetin and rutin flavonoids were also found in leaves and stalks of nettle by using modern spectroscopic (Nuclear magnetic resonance-NMR-, Mass Spectrometry-MS-, etc.) methods [13].

By using Supercritical Fluids Extraction (SFE) method in which liquid CO₂, different pressure, and temperature profiles were used, chlorophylll a, chlorophylll b, β-carotene, and lutein components were found in nettle leaves [14]. In another research chlorophylll a and chlorophyll b were isolated from nettles, which could be used as a coloring agents (E140) in the field of drug and food. The most efficient process was identified as 4-step extraction and nettle was firstly dried at 40°C, stored at dark place in plastic bags at 4°C [15].

Linoleic acid, palmitic acid, oleic acid, palmitoleic acid, stearic acid, gadoleic acid, and erucic acid were found in nettle roots analysis by using GC about fatty acids composition [16]. In a research (−)-epi catechin and (+)-catechin were found in nettle leaves by using RP-HPLC method and UV detector. Nettle has some antimicrobial effects as its phenolic contents. Nettle could inhibit Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes [8].

A plant mix “Antidiabetis” is used which could decrease blood glucose level and contains root and other parts of nettle in Croatia [17]. Nettle is also used as an antioxidant which could decrease muscular contraction as analgesic. It also shows antimicrobial effect and it has some benefit on gastric mucosal damage [18]. In household remedy nettle is used against muscle paralysis. According to this point, nettles effect on proteolytic activity of Botulinum neurotoxins was studied. Nettle leaves extract could inhibit Botulinum neurotoxin serotype A light chain protease activity without competition, but it has no effect on serotype B [19].

The aim of this study was to analyze phenolic component and antioxidant activity of nettle, which is grown in coastal part of Turkey (Mediterranean, Aegean, Black sea, and Marmara). In this paper the nettle was taken apart (root, stalk, and leaves), and these parts were analyzed separately. These analyses were done to indicate differences of nettles between regions and parts. The samples were coded as Mediterranean (01-Adana, 07-Antalya, 07F-Fethiye, 32-Isparta), Aegean (09Y-wild sample from Aydın, 09-Aydın, 20-Denizli, 35-Izmir, 45A-Alasehir, 45S-Salihli, 45T-Turgutlu, 48-Mugla), Black Sea (52-Ordu, 53-Rize, 55-Samsun, 61-Trabzon, 74-Bartın) and Marmara (16-Bursa, 41-Kocaeli) which are name of cities. On the other hand, the moisture content of nettles was also analyzed. Firstly nettles were extracted. Then, analysis was done by using extract of nettles. Phenolic profiles of nettle were determined by using HPLC (High-Performance Liquid Chromatography) method.

2. Materials and Methods

2.1. Materials

2.1.1. Nettle Samples

Samples were collected from Aegean, Black Sea, Marmara, and Mediterranean region. These samples were rapidly washed and dried. Then, they were separated into root, stalk and leaves. After this process, part of samples was kept in a zip lock bag at −20°C for inhibition of air contact before analysis.

2.1.2. Sample Preparation and Extraction

Required amount of the samples were taken and then whittle into small particles to increase extraction yield.

After trials of different types of extraction method [20, 21], 80% methanol-water mixture was used as an extraction liquid. 1 g samples were taken from each of samples. 10 mL extraction liquid was added into the samples and extracted during 1 h at 50°C. The extraction was done in closed vessels to inhibit loss of the extraction liquid. After extraction process, the eluate was filtered.

2.1.3. Chemical Materials

Standards-Gallic acid (Sigma, G7384), ferulic acid (Fluka, 42280), rutin (Sigma, R5143), myricetin (Sigma, M6760), syringic acid (Sigma, S6881), caffeic acid (Sigma, C0625), chlorogenic acid (Sigma, C3878), quercetin hydrate (Sigma, 337951), p-coumaric acid (Sigma, C9008), kaempferol (Sigma, K0133), catechin hydrate (Fluka, 22110), fumaric acid (Fluka, 47910), vanillic acid (Fluka, 94770), naringin (Sigma, N1376), ellagic acid (Sigma, E2250), and isorhamnetin (Fluka, 17794) were used.

Chemicals-Folin-ciocalteau phenol reactive (Sigma-Aldrich, E9252), acetic acid (Panreac, 361008), sodium carbonate (J.T. Baker, 2024), DPPH (2,2 diphenil, 1, picrylhydrazyl) (Sigma, D9132), and Chromatographically pure gradient solvent-Methanol (Labscan, A17C11) were used.

2.2. Method

2.2.1. Moisture Analysis

Moisture analysis of samples was done by using “Fruit and Vegetables Moisture Analysis” method [22].

According to method we have the following:(i)3 gr of nettle from each part (root, stalk, and leaves) was weighed (0,001 of accuracy).(ii)Firstly weighing bottles were kept in drying oven for 1 h at 103 ± 2°C and then samples were put in weighing bottles and kept in drying oven for 4 h at 103 ± 2°C.(iii)Then samples were taken out from drying oven and then kept in desiccators for 30 min to reach the room temperature.(iv)After being weighed, samples were put again in drying oven for 1 h to dry.(v)These drying periods (1 h) were done until reaching the constant weighing (7 h).(vi)The moisture contents of samples were measured by using the following formula: where is initial weigh of sample, is final weigh of sample.

2.2.2. Total Phenolic Content Analysis

Total phenolic content analysis of nettle extracts was done by using Folin-Ciocalteau (FC) method [23, 24]. The results were expressed as gallic acid equivalents in milligrams per gram of dry matter. The solutions was as follows:(i)FC Reactive,(ii)7% Na₂CO₃ solution,(iii)gallic acid standard solution (50–100–150–200–300 ppm) preparing by using 80% methanol.

According to the method, we have the following.(i)250 μL FC reactive was added into 50 μL nettle extract or standard solution.(ii)This mixture was stayed at room temperature in dark place for 5 min.(iii)At the end of this period, 750 μL 7% Na₂CO₃ solution was added. By this way, phenolics hydroxyl groups could give H to water.(iv)This mixture was completed to 5 mL with pure water.(v)Then, mixture was stayed at room temperature in dark place for 120 min for reaction.(vi)Samples and standards absorbance was measured at 760 nm.(vii)For blank solution instead of 50 μL extract, 80% methanol solution was added.(viii)Total phenolic content was measured with calibration curve by using gallic acid equivalent standards.

2.2.3. DPPH Antioxidant Capacity Analysis Method

Nettle extracts antioxidant capacity analysis was done by using DPPH radical degradation activity method [25].

According to the method, we have the following.(i)6 × 10-5 M (molar) DPPH radical was prepared daily by using pure methanol.(ii)2 μL methanolic DPPH solution was added into 100 μL sample extract or standard solution.(iii)This mixture was stayed in dark place for 20 min.(iv)At the end of this time, absorbance was measured at 515 nm.(v)Pure methanol was used as blank solution.(vi)For control solution, instead of 100 μL extract 100 μL pure water was used.(vii)Sample extract antioxidant capacities were measured by using calibration curve which was prepared by using different concentrations (10–100 ppm) of gallic acid solution.

2.2.4. HPLC Analysis of Phenolic Component

Qualitative and quantitative analyses of caffeic acid, vanillic acid, naringin, syringic acid, and ferulic acid, ellagic acid, myricetin, kaempferol, isorhamnetin, catechin, chlorogenic acid, p-coumaric acid, rutin, fumaric and gallic acid component in nettle samples were done by HPLC. HPLC analysis was done by using graded elution program. The elution program could be summarized as follows, 0–11 min, 100% A, 30–40 min, 35% A, and 65% B, and 42 min 100% A. A and B solvents were used as elution solvent. Solvent A consists of 2% acetic acid, 10% methanol, and 88% pure water, and B consists of 2% acetic acid, 90% methanol, and 8% pure water. The flow rate was 1 mL/min, temperature was 40°C, and the injection volume was 20 μL. On the other hand, according to maximum absorbance of standards, analysis was done at 254, 270, 280, and 370 nm wavelength. Some chromatogram samples are given at Figure 1. The standards chromatogram is given at Figure 2.

Figure 1: HPLC chromatogram of Antalya, Aydın, Bursa and Samsun root, stalk and leaves sample.

Figure 2: HPLC chromatogram of antioxidant standards. *1: fumaric acid; 2: gallic acid; 3: catechins; 4: vanillic acid; 5: caffeic + chlorogenic acid; 6: syringic acid; 7: p-coumaric acid; 8: ferulic; 9: naringin; 10: rutin; 11: ellagic; 12: myricetin; 13: quercetin; 14: kaempferol; 15: isorhamnetin.

In HPLC analysis of nettle samples, the methanolic extracts were used. Samples and standards were filtered from 0,45 μm Agilent micro filter, then were put into vials, and finally they were given to HPLC.

3. Results and Discussion

3.1. Moisture Contents

Moisture content analysis of root, stalk, leaves, and total of nettle was done. This moisture analysis was given in the table (Tables 1 and 2) as region. Analysis was duplicated. The results were given with standard deviations. The differences between samples were determined by using SPSS (v.17)/one-way-aNOVA/Duncan test .

Table 1: Moisture content, total phenolic content, and total antioxidant activity of total and root of fresh nettle.

Moisture %
FC (mg GAE/g DM)
DPPH (mg GAE/g DM)

Sample

M01
84,61 ± 0,05b
157,27 ± 5,33b
147,20 ± 2,09a

M07
74,93 ± 0,04c
307,10 ± 1,12a
241,63 ± 2,72a

M07F
74,03 ± 0,02c
344,12 ± 9,19a
247,16 ± 1,14a

M32
89,74 ± 0,02a
123,62 ± 1,68b
98,70 ± 1,35b

A09Y
89,80 ± 0,11a
243,68 ± 3,35c
51,44 ± 2,74b

A09
88,84 ± 0,18a
113,69 ± 2,87d
145,83 ± 0,39a

A20
88,88 ± 0,21a
730,65 ± 5,85d
52,78 ± 0,48b

A35
84,09 ± 0,06b
126,95 ± 1,07a
141,54 ± 1,14a

A45A
80,74 ± 0,00c
277,25 ± 3,65c
114,04 ± 1,30a

A45S
86,96 ± 0,21a,b
123,88 ± 0,98d
53,27 ± 0,31b

A45T
85,56 ± 0,15b
383,93 ± 3,96b
72,73 ± 0,35b

A48
89,08 ± 0,15a
66,97 ± 0,89d
114,11 ± 0,47a

B52
76,29 ± 0,11b
440,49 ± 16,06b
106,69 ± 0,91a

B53
78,45 ± 0,04a
233,93 ± 5,16b
235,38 ± 2,19a

B55
69,66 ± 0,05c
723,93 ± 37,56a
152,26 ± 1,05a

B61
78,64 ± 0,03a
353,92 ± 11,34b
224,87 ± 1,92a

B74
78,10 ± 0,02a,b
297,15 ± 2,90b
234,18 ± 1,16a

MS16
62,55 ± 0,05b
410,53 ± 3,21b
260,92 ± 1,01a

MS41
76,43 ± 0,03a
443,96 ± 6,33a
249,96 ± 1,91a

Root Sample

M01
87,89 ± 0,02b
70,47 ± 1,14c
150,85 ± 0,42a

M07
74,96 ± 0,08a
370,12 ± 0,93a
300,60 ± 1,83a

M07F
77,92 ± 0,01a
170,99 ± 4,39b
250,81 ± 0,95a

M32
89,79 ± 0,04b
20,44 ± 0,24c
130,01 ± 2,35a

A09Y
93,45 ± 0,05a
60,03 ± 0,34a
30,60 ± 1,16b

A09
91,31 ± 0,19a
20,12 ± 0,31b
50,60 ± 0,12b

A20
89,96 ± 0,10a,b
100,89 ± 0,18b
40,22 ± 0,20b

A35
86,36 ± 0,07b,c
90,86 ± 1,56b
170,42 ± 1,96a

A45A
84,35 ± 0,00c
40,49 ± 0,45b
90,74 ± 0,01a

A45S
89,46 ± 0,53a,b
90,94 ± 2,24b
40,31 ± 0,19b

A45T
86,39 ± 0,08b,c
90,35 ± 2,53b
60,97 ± 0,55b

A48
70,45 ± 0,26a
390,33 ± 9,60b
120,24 ± 0,51a

B52
71,38 ± 0,08a
790,21 ± 43,65a
140,58 ± 2,46a

B53
70,44 ± 0,01a
40,52 ± 0,13a
340,31 ± 1,31a

B55
73,56 ± 0,11a
1020,16 ± 69,40a
190,91 ± 2,35a

B61
78,09 ± 0,00a
270,62 ± 8,87a
350,72 ± 1,69a

B74
92,32 ± 0,01a
190,67 ± 0,36a
270,62 ± 0,67a

MS16
71,42 ± 0,03b
430,34 ± 6,20b
370,27 ± 0,11a

MS41
76,12 ± 0,00a
470,78 ± 0,46a
300,15 ± 2,13a

is representing Mediterranean, A is Aegean, B is Black Sea, and MS is Marmara region. The statistical analysis was done within Region.

Table 2: Moisture content, total phenolic content, and total antioxidant activity of stalk and leaves of fresh nettle.

Moisture %
FC (mg GAE/g DM)
DPPH (mg GAE/g DM)

Stalk Sample

M01
85,29 ± 0,02b
20,24 ± 0,68b,c
90,06 ± 0,52b

M07
84,11 ± 0,01b,c
80,46 ± 2,46b
300,61 ± 2,40a

M07F
81,32 ± 0,04c
150,35 ± 2,16a
180,39 ± 1,72a

M32
92,81 ± 0,02a
370,58 ± 8,51c
230,39 ± 5,16a

A09Y
93,40 ± 0,20a
10,08 ± 0,06e
30,31 ± 0,19b

A09
91,17 ± 0,30b,c
60,71 ± 1,34c
50,77 ± 0,15b

A20
91,62 ± 0,03a,b
150,07 ± 2,31b
40,85 ± 0,48b

A35
87,85 ± 0,02d
100,42 ± 0,64a
140,15 ± 0,91a

A45A
84,66 ± 0,05e
70,49 ± 1,60b,c
130,82 ± 0,42a

A45S
90,71 ± 0,01b,c
40,66 ± 2,03c,d
40,29 ± 0,17b

A45T
89,15 ± 0,09c,d
60,25 ± 0,59c,d
70,19 ± 0,07b

A48
77,84 ± 0,12c,d
30,28 ± 0,96d,e
30,71 ± 0,68b

B52
79,41 ± 0,08a
160,72 ± 3,47a
90,41 ± 0,15a

B53
79,86 ± 0,13a
320,85 ± 4,53a
290,63 ± 3,65a

B55
80,36 ± 0,01a
270,62 ± 28,61a
90,35 ± 0,10a

B61
77,48 ± 0,05a
260,61 ± 6,84a
240,45 ± 3,04a

B74
89,41 ± 0,03a
480,30 ± 3,33a
300,93 ± 2,01a

MS16
43,78 ± 0,04b
120,71 ± 3,38a
240,86 ± 1,48a

MS41
78,91 ± 0,05a
100,90 ± 4,79b
320,99 ± 3,44a

Leaves Sample

M01
86,63 ± 0,02b
320,88 ± 12,39a
110,39 ± 4,14a

M07
80,65 ± 0,11c
400,83 ± 0,26a
190,04 ± 1,15a

M07F
65,72 ± 0,02c
490,80 ± 14,67a
170,79 ± 0,07a

M32
62,84 ± 0,01a
330,20 ± 4,12a
70,54 ± 1,20b

A09Y
82,57 ± 0,10a
330,61 ± 0,38d
80,53 ± 6,89a

A09
84,04 ± 0,04a
190,01 ± 4,74d,e
320,38 ± 0,89a

A20
85,07 ± 0,48a
220,40 ± 2,22d,e
60,77 ± 0,75a

A35
78,07 ± 0,09a,b
1941,00 ± 15,06a
100,90 ± 0,53a

A45A
80,59 ± 0,12b
150,52 ± 1,41c
80,17 ± 0,49a

A45S
80,72 ± 0,01a
1000,61 ± 9,06d,e
100,56 ± 0,34a

A45T
81,13 ± 0,25a
670,40 ± 7,78b
170,28 ± 3,81a

A48
73,22 ± 0,00a
290,50 ± 0,45e
70,96 ± 0,06a

B52
84,56 ± 0,14a
370,54 ± 1,06b
80,02 ± 0,11a

B53
58,68 ± 0,11a
340,43 ± 10,82b
60,62 ± 1,60a

B55
81,99 ± 0,00b
881,00 ± 14,67a
160,43 ± 0,71a

B61
78,74 ± 0,06a
530,52 ± 18,32b
70,29 ± 1,02a

B74
85,51 ± 0,04a
220,49 ± 5,01b
110,71 ± 0,79a

MS16
72,44 ± 0,09b
680,55 ± 0,04b
160,14 ± 1,44a

MS41
74,25 ± 0,04a
760,20 ± 13,75a
110,85 ± 0,15a

is representing Mediterranean, A is Aegean, B is Black Sea, and MS is Marmara region. The statistical analysis was done within region.

According to scientific studies, moisture analysis was done to a nettle tea bag which includes all parts of nettle and as a result its moisture content was 6,3% [26]. In another study about nettle leaves which was collected from Macedonian, moisture content was 8,3% [1]. In another research about nettles (root, stalk and leaves), moisture contents of nettles root, stalk and leaves were 80,01%, 88,88%, and 78,67%, respectively [27].

In our study, average moisture content of nettles in total samples was 80,94%, root samples 81.87%, stalk samples 83,11%, and leaves samples 77,75%. The highest moisture content was 09 and the lowest one was 16 in total. In root the highest moisture content was 09Y and the lowest one was 53. In stalk the highest moisture content was 09Y and the lowest one was 16. In leaves the highest moisture content was 32 and the lowest one was 07F. On the other hand, the moisture content was different among the regions. As a general comment moisture content could be ranged like stalk > root > leaves.

Nettle samples, which were used in this study, are fresh samples that could be a reason of the high moisture content results.

3.2. Total Phenolic Content Analysis of Nettle

According to the analysis result (Tables 1 and 2), total phenolic content of nettle was indicated with FC method, which is a wide used method. Nettles (total, root, stalk, and leaves) were analyzed and this analysis result was given at 760 nm wavelengths. SPSS (v.17) statistical program, one-way-aNOVA/Duncan test was used to indicate the difference among samples. As a general comparison among samples, total phenolic content of nettle could be ranged as leaves > root > stalk.

According to a scientific research, total phenolic content analyses of nettle parts (root, stalk, and leaves) were done by using FC method. The results were given as mg Gallic Acid Equivalent (GAE)/g Dry Matter (DM). In this paper, the results were given as root 7,82, stalk 9,91, and leaves 7,62 mg GAE/g DM [27]. In comparison, our nettle samples total phenolic content is higher than this research results. In a research about phenolic analysis of nettle as cultivated and wild, total phenolic contents of nettles were given for stalk of nettle 28,6% in cultivated and 24,4% in wild samples, while 71,5% and 76,5% for leaves of nettle, respectively [28].

In another research, total phenolic content of nettle tea bag which includes all parts of nettle (root, stalk, and leaves) was 2,5 mg GAE/g DM [26].

3.3. Total Antioxidant Activity Analysis

Antioxidant activity analysis of nettle parts (root, stalk, and leaves) was done by using DPPH antioxidant activity method. The analysis result was given at 515 nm wavelengths. The results are given in Tables 1 and 2. SPSS (v.17) statistical program, one-way-aNOVA/Duncan test was used to indicate the difference among samples.

In a research related to nettle parts (root, stalk, and leaves), antioxidant activities were given as root 9,86, stalk 37,56, and leaves 76,06 mg GAE/g DM [27]. On the other hand, in another study about herbal tea, a nettle tea bag (include root, stalk, and leaves of nettle) total antioxidant activity was 2,5 mg GAE/g DM [26]. The process of nettle to prepare tea could be the reason of the lower antioxidant activity instead of fresh nettle parts. In a general perspective, antioxidant activity could be ranged as root > stalk > leaves. The 41 sample has the highest total antioxidant activity, and the 20 sample has the lowest one. In root sample, the highest one was 16 and the lowest one was 20. In stalk sample, the highest one was 41 and the lowest one was 09Y. In leaves sample, the highest one was 09 and the lowest one was 53.

The total antioxidant activity of nettle parts (root, stalk and leaves) was analyzed by using DPPH method. While the results were compared with literatures total antioxidant activity of fresh nettle was higher than the others (nettle tea, dry nettle leaves). According to the DPPH analysis results statistical discrepancy were observed between Mediterranean, Aegean, Black Sea, Marmara Region, root, stalk and leaves. On the other hand, there was no statistical discrepancy between Mediterranean leaves sample, Black Sea root, and stalk sample.

3.4. HPLC Analyses of Phenolic Component of Nettle

The total phenolic content and antioxidant activity of nettle samples which were collected from different regions and cities of Turkey were diverse between regions, cities, root, stalk and leaves. Nettles (total, root, stalk, and leaves) were analyzed and the results are given in Tables 3, 4, and 5. 16 antioxidant standards were used for identification of phenolic component of nettle sample in this research. Samples methanolic extracts were analyzed with these standards by HPLC-DAD. SPSS (v.17) statistical program, one-way-aNOVA/Duncan test was used to indicate the difference among samples.

Table 3: Phenolic content of fresh nettle roots.

(a)

Sample
Gallic
Syringic
Myricetin
Quercetin
Kaempferol
Fumaric
Vanillic

M01
—
—
1,45 ± 0,13a,b
0,74 ± 0,02a
0,49 ± 0,11a
—
—

M07
—
—
0,64 ± 0,014b
0,84 ± 0,10a
0,47 ± 0,19a
—
—

M07F
—
—
2,18 ± 0,079a,b
0,73 ± 0,01a
0,52 ± 0,52a
—
—

M32
—
—
4,90 ± 1,83a
0,75 ± 0,01a
0,46 ± 0,01a
—
—

A09Y
—
0,00 ± 0,00b
0,70 ± 0,08b
0,38 ± 0,54b
0,44 ± 0,01a
—
—

A09
—
0,00 ± 0,00b
1,13 ± 0,34b
0,78 ± 0,08a,b
0,25 ± 0,35a
—
—

A20
—
4,31 ± 2,74a
21,93 ± 3,69a
0,77 ± 0,07a,b
0,46 ± 0,01a
—
—

A35
—
0,31 ± 0,43b
0,89 ± 0,08b
0,76 ± 0,02a,b
0,24 ± 0,34a
—
—

A45A
—
0,00 ± 0,00b
0,64 ± 0,01b
0,78 ± 0,09a,b
0,45 ± 0,03a
—
—

A45S
—
0,00 ± 0,00b
0,75 ± 0,02b
0,73 ± 0,01a,b
0,22 ± 0,32a
—
—

A45T
—
0,00 ± 0,00b
4,36 ± 2,76b
1,15 ± 0,33a
0,51 ± 0,09a
—
—

A48
—
0,00 ± 0,00b
0,34 ± 0,48b
0,71 ± 0,01a,b
0,22 ± 0,31a
—
—

B52
—
—
0,64 ± 0,02a
0,73 ± 0,00a
0,23 ± 0,32a
—
—

B53
—
—
0,49 ± 0,69a
0,76 ± 0,01a
0,24 ± 0,33a
—
—

B55
—
—
0,71 ± 0,06a
0,84 ± 0,14a
0,66 ± 0,09a
—
—

B61
—
—
0,69 ± 0,01a
0,76 ± 0,05a
0,51 ± 0,08a
—
—

B74
—
—
0,69 ± 0,02a
0,74 ± 0,01a
0,46 ± 0,02a
—
—

MS16
—
2,99 ± 0,83a
0,00 ± 0,00b
0,38 ± 0,55a
0,24 ± 0,33a
6,28 ± 8,88a
—

MS41
—
0,00 ± 0,00a
0,66 ± 0,01a
0,75 ± 0,01a
0,54 ± 0,09a
0,00 ± 0,00a
—

is representing Mediterranean, A is Aegean, B is Black Sea, and MS is Marmara Region. The statistical analyses were done within region.

(b)

Sample
Rutin
Ellagic
Isorhamnetin
Catechin
Caffeic + chlorogenic
p-coumaric
Ferulic
Naringin

M01
6,38 ± 4,10b
6,50 ± 1,62b
—
—
—
1,86 ± 0,32a
1,15 ± 1,14b
2,66 ± 0,23b

M07
3,93 ± 0,80b
1,86 ± 1,86b
—
—
—
3,37 ± 2,01a
10,18 ± 1,01a
37,97 ± 5,67a

M07F
59,60 ± 20,79a
2,03 ± 2,03b
—
—
—
5,29 ± 2,38a
27,43 ± 10,52a
5,61 ± 1,61b

M32
0,00 ± 0,00b
26,81 ± 0,18a
—
—
—
0,00 ± 0,00a
0,39 ± 0,39b
0,00 ± 0,00b

A09Y
0,00 ± 0,00b
0,00 ± 0,00b
0,00 ± 0,00b
—
—
1,94 ± 0,33b
2,57 ± 0,41b
0,00 ± 0,00d

A09
0,92 ± 0,31a,b
3,51 ± 0,50a,b
3,42 ± 4,83b
—
—
3,16 ± 1,19a,b
0,00 ± 0,00b
0,49 ± 0,69c,d

A20
0,90 ± 0,27a,b
3,95 ± 5,59a,b
0,00 ± 0,00b
—
—
8,42 ± 5,60a
29,34 ± 3,06a
0,46 ± 0,06a

A35
0,73 ± 1,03a,b
0,00 ± 0,00b
84,85 ± 4,04a
—
—
3,83 ± 0,33a,b
0,00 ± 0,00b
2,52 ± 1,45b,c,d

A45A
0,49 ± 0,69a,b
0,00 ± 0,00b
0,00 ± 0,00b
—
—
5,06 ± 0,75a,b
0,00 ± 0,00b
3,10 ± 0,21b,c

A45S
0,00 ± 0,00b
0,00 ± 0,00b
27,63 ± 13,48b
—
—
2,53 ± 0,25a,b
0,00 ± 0,00b
0,00 ± 0,00d

A45T
0,98 ± 0,58a,b
12,92 ± 13,09a
0,00 ± 0,00b
—
—
3,00 ± 4,24a,b
0,99 ± 1,41b
0,87 ± 1,23c,d

A48
1,53 ± 0,52a
0,00 ± 0,00b
23,70 ± 33,52b
—
—
2,49 ± 0,69a,b
1,21 ± 1,71b
5,04 ± 2,45a,b

B52
1,71 ± 0,35a
10,74 ± 1,81a
17,49 ± 17,24a
—
—
3,71 ± 1,64a
4,26 ± 2,87a
9,84 ± 12,01a

B53
1,39 ± 0,64a
3,94 ± 0,05a,b
5,65 ± 8,00a
—
—
5,02 ± 0,83a
3,74 ± 4,80a
19,72 ± 11,27a

B55
4,20 ± 1,25a
3,59 ± 5,09a,b
15,29 ± 11,32a
—
—
8,34 ± 0,15a
11,27 ± 0,52a
20,49 ± 0,93a

B61
6,43 ± 5,60a
1,39 ± 1,97b
1,37 ± 1,93a
—
—
4,16 ± 1,77a
3,83 ± 1,05a
14,50 ± 4,68a

B74
4,53 ± 4,48a
5,39 ± 1,92a,b
0,37 ± 0,52a
—
—
6,99 ± 6,09a
7,06 ± 5,01a
10,15 ± 6,11a

MS16
2,43 ± 2,37a
2,01 ± 2,83a
5,47 ± 7,74a
—
—
3,23 ± 0,52a
4,23 ± 1,25a
13,56 ± 5,64a

MS41
2,55 ± 3,07a
2,03 ± 2,87a
10,91 ± 15,43a
—
—
4,76 ± 1,77a
6,05 ± 2,46a
17,31 ± 1,79a

is representing Mediterranean, A is Aegean, B is Black Sea, and MS is Marmara Region. The statistical analyses were done within region.

Table 4: Phenolic content of fresh nettle stalks.

(a)

Sample
Gallic
Syringic
Myricetin
Quercetin
Kaempferol
Fumaric
Vanillic

M01
—
0,00 ± 0,00a
1,15 ± 0,18a,b
0,72 ± 0,01a
0,24 ± 0,24a
6,18 ± 6,18a
—

M07
—
0,00 ± 0,00a
0,75 ± 0,08b
0,37 ± 0,37a
0,38 ± 0,38a
11,66 ± 0,99a
—

M07F
—
2,63 ± 1,94a
1,57 ± 0,21a
1,02 ± 0,05a
0,24 ± 0,24a
9,17 ± 1,32a
—

M32
—
0,00 ± 0,00a
0,66 ± 0,01b
0,76 ± 0,05a
0,26 ± 0,26a
2,09 ± 2,96a
—

A09Y
—
0,00 ± 0,00a
0,50 ± 0,72b
0,36 ± 0,51a
0,48 ± 0,05a
—
0,00 ± 0,00b

A09
—
0,00 ± 0,00a
0,76 ± 0,16b
0,75 ± 0,01a
0,44 ± 0,01a
—
0,00 ± 0,00b

A20
—
1,20 ± 1,69a
8,96 ± 11,65a
0,77 ± 0,05a
0,51 ± 0,05a
—
0,00 ± 0,00b

A35
—
2,49 ± 1,22a
1,26 ± 0,01b
0,78 ± 0,07a
0,45 ± 0,01a
—
39,46 ± 18,00a

A45A
—
0,00 ± 0,00a
4,61 ± 5,61b
0,77 ± 0,05a
0,45 ± 0,01a
—
0,00 ± 0,00b

A45S
—
49,28 ± 69,69a
1,85 ± 1,10b
4,54 ± 5,33a
0,31 ± 0,44a
—
0,00 ± 0,00b

A45T
—
0,00 ± 0,00a
3,70 ± 2,59b
0,79 ± 0,03a
0,25 ± 0,35a
—
0,00 ± 0,00b

A48
—
0,00 ± 0,00a
3,02 ± 2,14b
0,73 ± 0,01a
0,49 ± 0,04a
—
0,00 ± 0,00b

B52
—
—
0,78 ± 0,14a
0,74 ± 0,03a
0,63 ± 0,26b
—
—

B53
—
—
0,85 ± 0,22a
0,77 ± 0,02a
0,22 ± 0,32b
—
—

B55
—
—
0,76 ± 0,10a
0,73 ± 0,01a
0,52 ± 0,12b
—
—

B61
—
—
0,93 ± 0,40a
0,73 ± 0,03a
1,21 ± 0,20a
—
—

B74
—
—
0,67 ± 0,03a
0,74 ± 0,01a
0,47 ± 0,39b
—
—

MS16
—
1,12 ± 1,32a
0,32 ± 0,46a
0,73 ± 0,01a
—
8,18 ± 0,85a
—

MS41
—
0,00 ± 0,00a
0,71 ± 0,07a
0,32 ± 0,01b
—
0,00 ± 0,00b
—

is representing Mediterranean, A is Aegean, B is Black Sea, and MS is Marmara Region. The statistical analyses were done within region.

(b)

Sample
Rutin
Ellagic
Isorhamnetin
Catechin
Caffeic + chlorogenic
p-coumaric
Ferulic
Naringin

M01
29,62 ± 26,66a
2,93 ± 2,92a
0,94 ± 0,94a,b
—
25,91 ± 23,73a
3,87 ± 2,55a
20,82 ± 1,37a,b
3,79 ± 0,77b

M07
7,72 ± 3,17a
1,96 ± 1,96a
0,00 ± 0,00b
—
0,00 ± 0,00a
0,00 ± 0,00a
0,00 ± 0,00b
8,81 ± 0,25a

M07F
6,41 ± 1,66a
3,82 ± 0,19a
3,14 ± 0,78a
—
17,18 ± 1,75a
8,07 ± 1,59a
44,50 ± 22,24a
6,05 ± 2,29a,b

M32
33,65 ± 33,65a
0,00 ± 0,00a
0,00 ± 0,00b
—
0,00 ± 0,00a
0,00 ± 0,00a
0,00 ± 0,00b
0,00 ± 0,00c

A09Y
0,00 ± 0,00b
0,00 ± 0,00b
0,00 ± 0,00a
—
—
1,64 ± 0,57b
4,44 ± 0,58b
0,00 ± 0,00c

A09
1,20 ± 0,24b
1,54 ± 2,17b
0,00 ± 0,00a
—
—
2,41 ± 0,21b
0,03 ± 0,05b
0,21 ± 0,30c

A20
1,61 ± 0,15b
1,90 ± 2,69b
5,81 ± 8,22a
—
—
0,00 ± 0,00b
0,12 ± 0,16b
0,00 ± 0,00c

A35
0,00 ± 0,00b
3,62 ± 0,04b
46,49 ± 0,10a
—
—
22,04 ± 6,32a
0,00 ± 0,00b
12,08 ± 0,24a

A45A
0,35 ± 0,50b
0,00 ± 0,00b
0,00 ± 0,00a
—
—
2,70 ± 1,65b
0,00 ± 0,00b
1,27 ± 0,77b,c

A45S
3,34 ± 4,72b
4,96 ± 7,01b
19,39 ± 21,21a
—
—
7,18 ± 10,16b
11,20 ± 15,84a,b
3,95 ± 3,38b

A45T
19,31 ± 0,14a
29,39 ± 1,82a
0,00 ± 0,00a
—
—
1,50 ± 2,13b
24,16 ± 11,28b
1,62 ± 2,29b,c

A48
1,76 ± 2,48b
2,35 ± 3,33b
34,96 ± 49,45a
—
—
2,79 ± 0,52a,b
8,77 ± 0,78a,b
1,08 ± 0,63b,c

B52
3,07 ± 1,85a
6,07 ± 3,80a
0,51 ± 0,71b
—
—
6,45 ± 1,79a
5,53 ± 0,00a
13,04 ± 15,16b

B53
1,32 ± 0,39a
4,06 ± 2,08a
11,03 ± 2,51a,b
—
—
8,88 ± 1,47a
2,04 ± 2,88a
12,91 ± 0,01b

B55
0,79 ± 1,12a
5,38 ± 2,16a
16,91 ± 15,55a,b
—
—
3,29 ± 0,34a
1,66 ± 1,95a
16,42 ± 5,41b

B61
9,32 ± 8,65a
7,85 ± 2,38a
30,02 ± 7,46a
—
—
9,69 ± 3,75a
5,68 ± 1,37a
63,19 ± 8,68a

B74
9,76 ± 2,49a
5,68 ± 2,76a
0,00 ± 0,00b
—
—
3,08 ± 0,77a
5,84 ± 1,46a
32,00 ± 10,27b

MS16
2,03 ± 0,75a
—
—
—
—
1,48 ± 0,11a
0,86 ± 1,21a
7,19 ± 0,22a

MS41
4,38 ± 3,72a
—
—
—
—
3,78 ± 1,19a
0,00 ± 0,00a
22,48 ± 12,20a

is representing Mediterranean, A is Aegean, B is Black Sea and MS is Marmara Region. The statistical analyses were done within Region.

Table 5: Phenolic content of fresh nettle leaves.

(a)

Sample
Gallic
Syringic
Myricetin
Quercetin
Kaempferol
Fumaric
Vanillic

M01
—
0,00 ± 0,00a
1,15 ± 0,33b
0,96 ± 0,13b
0,61 ± 0,15a
—
26,15 ± 36,98a

M07
—
23,10 ± 32,67a
0,75 ± 1,48a,b
1,37 ± 0,03a,b
0,60 ± 0,01a
—
0,00 ± 0,00a

M07F
—
2,78 ± 3,93a
1,57 ± 0,55a
1,78 ± 0,17a
0,61 ± 0,17a
—
0,42 ± 0,59a

M32
—
0,00 ± 0,00a
0,65 ± 0,01b
1,23 ± 0,50a,b
0,49 ± 0,06a
—
0,00 ± 0,00a

A09Y
—
16,20 ± 2,07b
0,79 ± 0,22b
0,91 ± 0,11a
5,96 ± 2,05a
—
0,00 ± 0,00b

A09
—
341,68 ± 21,04a
2,08 ± 1,45b
1,19 ± 0,42a
1,68 ± 0,03b
—
40,68 ± 10,78b

A20
—
0,00 ± 0,00b
1,91 ± 0,73b
3,48 ± 2,30a
0,75 ± 0,13b
—
0,00 ± 0,00b

A35
—
0,00 ± 0,00b
1,67 ± 0,35b
1,16 ± 0,16a
0,68 ± 0,32b
—
295,21 ± 82,59a

A45A
—
0,00 ± 0,00b
1,20 ± 0,33b
1,38 ± 0,82a
0,93 ± 0,39b
—
0,00 ± 0,00b

A45S
—
0,00 ± 0,00b
1,03 ± 0,28b
1,38 ± 0,24a
2,15 ± 2,16b
—
20,85 ± 4,61b

A45T
—
0,00 ± 0,00b
6,39 ± 0,56a
2,95 ± 2,23a
0,59 ± 0,16b
—
0,00 ± 0,00b

A48
—
4,05 ± 5,73b
1,12 ± 0,63b
1,77 ± 0,12a
1,34 ± 1,30b
—
0,00 ± 0,00b

B52
—
7,80 ± 2,20a
0,82 ± 0,16a
1,30 ± 0,27a,b
0,97 ± 0,76a
—
—

B53
—
5,04 ± 7,13a
0,91 ± 0,36a
1,28 ± 0,15a,b
0,74 ± 0,18a
—
—

B55
—
0,00 ± 0,00a
1,26 ± 0,76a
1,37 ± 0,43a
1,07 ± 0,03a
—
—

B61
—
5,24 ± 7,42a
0,75 ± 0,13a
0,85 ± 0,06a,b
0,76 ± 0,47a
—
—

B74
—
7,49 ± 5,46a
1,26 ± 0,25a
0,79 ± 0,02b
1,33 ± 0,27a
—
—

MS16
—
48,99 ± 12,66a
0,88 ± 0,29a
0,96 ± 0,20a
0,85 ± 0,32a
10,93 ± 10,86a
—

MS41
—
46,92 ± 5,20a
2,18 ± 0,83a
0,84 ± 0,15b
1,69 ± 1,10a
0,00 ± 0,00a
—

is representing Mediterranean, A is Aegean, B is Black Sea and MS is Marmara Region. The statistical analyses were done within Region.

(b)

Sample
Rutin
Ellagic
Isorhamnetin
Catechin
Caffeic + chlorogenic
p-coumaric
Ferulic
Naringin

M01
21,85 ± 23,91c
4,65 ± 6,57a,b
7,00 ± 9,90a
—
60,89 ± 1,87a
3,34 ± 0,01b
1,66 ± 1,02a
7,35 ± 2,78a

M07
96,67 ± 10,09b
9,62 ± 1,87a,b
12,42 ± 17,16a
—
0,00 ± 0,00b
1,87 ± 0,44b
11,61 ± 16,42a
9,41 ± 0,46a

M07F
191,07 ± 35,23a
15,76 ± 4,11a
8,34 ± 11,53a
—
80,38 ± 21,43a
9,25 ± 3,72a
7,14 ± 2,86a
12,79 ± 5,21a

M32
0,80 ± 0,26c
0,00 ± 0,00b
0,00 ± 0,00a
—
4,77 ± 6,75b
0,00 ± 0,00b
0,00 ± 0,00a
11,62 ± 0,31a

A09Y
2,74 ± 1,14b
8,18 ± 11,56b
11,37 ± 16,09a,b
—
0,00 ± 0,00b
3,42 ± 1,12a
8,69 ± 7,73b
8,73 ± 4,59b

A09
60,74 ± 26,33a
29,91 ± 25,44a,b
0,00 ± 0,00b
—
61,63 ± 18,81a
2,94 ± 0,19a
20,36 ± 1,76a
34,04 ± 13,58a

A20
0,97 ± 0,25b
5,73 ± 0,47b
7,71 ± 0,36a,b
—
26,13 ± 36,96b
1,31 ± 1,85a
0,23 ± 0,33c
0,00 ± 0,00b

A35
5,78 ± 1,33b
17,44 ± 1,97a,b
32,81 ± 14,81a
—
0,00 ± 0,00b
12,80 ± 18,11a
1,06 ± 1,49c
24,08 ± 2,37a

A45A
0,71 ± 0,49b
5,53 ± 1,84b
12,45 ± 5,09a,b
—
0,00 ± 0,00b
2,25 ± 0,22a
0,00 ± 0,00c
2,92 ± 0,61b

A45S
2,39 ± 1,06b
6,06 ± 1,48b
24,55 ± 21,45a,b
—
0,00 ± 0,00b
0,00 ± 0,00a
0,00 ± 0,00c
2,68 ± 0,16b

A45T
6,64 ± 3,18b
42,35 ± 5,88a
0,70 ± 0,99b
—
0,00 ± 0,00b
4,28 ± 1,42a
3,36 ± 1,59b,c
6,26 ± 5,57b

A48
0,84 ± 1,19b
11,39 ± 16,11b
0,00 ± 0,00b
—
0,00 ± 0,00b
4,87 ± 0,14a
0,52 ± 0,22c
4,77 ± 0,13b

B52
7,29 ± 4,32a
10,73 ± 10,47a
46,91 ± 58,43a
—
—
4,70 ± 1,68a
2,54 ± 0,05a
4,05 ± 1,03a

B53
4,01 ± 0,39a,b
4,63 ± 1,91a
12,56 ± 10,05a
—
—
4,71 ± 1,51a
2,72 ± 3,06a
2,85 ± 1,12a

B55
3,85 ± 0,71a,b
9,14 ± 3,29a
11,52 ± 11,00a
—
—
4,12 ± 2,82a
3,35 ± 3,77a
5,29 ± 2,63a

B61
0,62 ± 0,88b
7,68 ± 3,80a
0,00 ± 0,00a
—
—
4,68 ± 4,75a
5,64 ± 5,55a
17,15 ± 18,45a

B74
8,27 ± 1,75a
10,21 ± 6,05a
10,04 ± 14,20a
—
—
5,12 ± 0,55a
0,28 ± 0,39a
9,48 ± 4,41a

MS16
20,43 ± 4,91a
5,14 ± 1,28a
—
—
—
1,54 ± 0,37a
2,43 ± 3,43a
10,24 ± 0,57a

MS41
16,97 ± 2,85a
17,78 ± 1,71a
—
—
—
1,80 ± 2,54a
4,71 ± 2,42a
17,65 ± 1,75a

is representing Mediterranean, A is Aegean, B is Black Sea and MS is Marmara Region. The statistical analyses were done within Region.

According to the result, total phenolic components of nettle samples were considerably high by comparison of other researches. The analysis results have statistical discrepancy between regions, cities, and parts of nettles (root, stalk, and leaves). Nettle parts (root, stalk, and leaves) phenolic content analysis was done qualitative and quantitative by using HPLC-DAD.

According to the results, there were not any gallic acid, syringic, fumaric, vanillic, isorhamnetin, catechin, caffeic, and chlorogenic acid in the root samples from Mediterranean region, but there were myricetin, rutin, ellagic acid, ferulic, and naringin. These standards have statistical discrepancy. There were not any gallic acid, vanillic, and catechin in stalk samples, but there were myricetin, isorhamnetin, ferulic and naringin. These standards have statistical discrepancy. There were not gallic acid, fumaric, and catechin in leaves samples, but there were myricetin, quercetin, rutin, ellagic, caffeic, and chlorogenic acid. These standards have statistical discrepancy.

There were not gallic acid, fumaric, vanillic, catechin, caffeic, and chlorogenic acid in root samples from Aegean region; other standards were found in these samples. There were not gallic acid, fumaric, catechin, caffeic, and chlorogenic acid, but there were syringic, quercetin, kaempferol, and isorhamnetin in stalk samples. There were not gallic acid, fumaric, and catechin, but there were quercetin and p-coumaric acid in leaves samples.

There were not gallic acid, syringic, fumaric, vanillic, catechin, caffeic, and chlorogenic acid, but other standards were found in root samples from Black Sea region. There were not gallic acid, syringic, fumaric, vanillic, catechin, caffeic, and chlorogenic acid in stalk samples, but there were kaempferol, isorhamnetin, and naringin. There were not gallic acid, fumaric, vanillic, catechin, caffeic, and chlorogenic acid in leaves samples, but there was statistical discrepancy in quercetin and fumaric acid.

There were not gallic acid, vanillic, catechin, caffeic, and chlorogenic acid in Marmara region root samples, but other standards were found. There were not gallic acid, kaempferol, vanillic, catechin, ellagic, isorhamnetin, caffeic, and chlorogenic acid in stalk samples, but other standards were found. There were not gallic acid, vanillic, isorhamnetin, catechin, caffeic, and chlorogenic acid in leaves samples, but other standards were found.

By comparison of root samples, p-coumaric, kaempferol, and quercetin have not statistical discrepancy. On the other hand, there were no gallic acid, fumaric, vanillic, catechin, caffeic, and chlorogenic acid. By comparison of stalk samples, syringic, myricetin, quercetin, kaempferol, and rutin have not statistical discrepancy. On the other hand, there were not gallic acid, vanillic acid, catechin, caffeic, and chlorogenic acid. By comparison of leaves samples, p-coumaric, isorhamnetin, and quercetin have not statistical discrepancy. On the other hand, there were not gallic acid, fumaric acid, and catechin.

In a research about cultivated and wild nettle samples phenolic profile and HPLC analysis, caffeic acid derivative, chlorogenic acid, 2-O-caffeoylmalic acid, rutin, quercetin 3-O-glucoside, kaempferol 3-O-rutinoside, and isorhamnetin 3-O-rutinoside were found in cultivated leaves samples. Caffeic acid derivative, p-coumaric acid, caffeoylquinic acid, chlorogenic acid, rutin, quercetin 3-O-glucoside, kaempferol 3-O-rutinoside, and isorhamnetin 3-O-rutinoside, peonidin 3-O-rutinoside, peonidin 3-O-(6′′-O-p-coumaroylglucoside), and rosinidin 3-O-rutinoside were found in cultivated stalk samples. Caffeic acid derivative, p-coumaric acid, chlorogenic acid, 2-O-caffeoylmalic acid, rutin, quercetin 3-O-glucoside, kaempferol 3-O-rutinoside, isorhamnetin 3-O-rutinoside were found in wild leaves samples. Finally, caffeic acid derivative, p-coumaric acid, caffeoylquinic acid, chlorogenic acid, rutin, quercetin 3-O-glucoside, kaempferol 3-O-rutinoside, isorhamnetin 3-O-rutinoside, peonidin 3-O-rutinoside, rosinidin 3-O-rutinoside were found in wild stalk samples [28]. According to our results, there were not any gallic acid, vanillic, and catechin in stalk samples, but there were myricetin, isorhamnetin, ferulic, and naringin in the stalk samples from Mediterranean region. There were not gallic acid, fumaric, catechin, caffeic, and chlorogenic acid, but there were syringic, quercetin, kaempferol, and isorhamnetin in stalk samples from Agean region. There were not gallic acid, syringic, fumaric, vanillic, catechin, caffeic, and chlorogenic acid in stalk samples from Black Sea region, but there were kaempferol, isorhamnetin, and naringin. There were not gallic acid, kaempferol, vanillic, catechin, ellagic, isorhamnetin, caffeic, and chlorogenic acid in stalk samples from Marmara region, but other standards were found. There were not gallic acid, fumaric, and catechin, but there were myricetin, quercetin, rutin, ellagic, caffeic, and chlorogenic acid in leaves samples from Mediterranean region. There were not gallic acid, fumaric, and catechin, but there were quercetin, and p-coumaric acid in leaves samples from Agean region. There were not gallic acid, fumaric, vanillic, catechin, caffeic, and chlorogenic acid in leaves samples from Black Sea region, but there was statistical discrepancy in quercetin, and fumaric acid. There were not gallic acid, vanillic, isorhamnetin, catechin, caffeic, and chlorogenic acid in leaves samples from Marmara region, but other standards were found.

In another research, some kinds of plants HPLC-MS analysis were done. One of these plants was nettle, and according to the results nettles included 18 ppm quercetin, 13 ppm myricetin, and 7,7 ppm kaempferol [29]. In comparison to our study, root, stalk, and leaves parts of nettle have lower phenolics components than those in this study, as seen in Tables 1 and 2.

4. Conclusions

Nettle is a plant easy to grow. Nettle is rich of chemical component and composition. It is as widely used from cosmetics to food. It is widely used plant in complementary and alternative treatment method (CTM and ATM) as cancer, and so forth. It is thought that nettle has these positive properties as its phenolic contents. Also, these plant parts (root, stalk, and leaves) have different phenolic composition and contents. The nettle part content differences indicate that different parts could be used for different cancers in ATM. Because of that reason, the phenolic compounds and contents of nettle parts were tried to identify. By this way, different kinds of phenolic components of nettles part (root, stalk, and leaves) indicate different usage area to the plant parts. Phenolic content analyses were done by using nettle parts (root, stalk and leaves) and the nettles were collected from different regions of Turkey (Aegean, Black Sea, Mediterranean, Marmara), according to these properties of nettle.

Nettle prefers nutrient riches and lighted places, hot and mild climate. Therefore, the higher total phenolic contents and antioxidant activities of nettles and roots of nettles were found in Marmara and Black Sea region in this research, while the higher moisture contents of nettles, roots, and stalks of nettles were found in Aegean and Mediterranean region. The higher moisture contents of leaves were found in Aegean and Black Sea region. The higher total phenolic contents of stalk samples was found in Black Sea and Mediterranean region, while the higher total antioxidant activities of stalk samples were found in Marmara and Black Sea region. The higher total phenolic contents of leaves samples were found in Marmara and Aegean region, while the higher total antioxidant activity of leaves samples were found in Marmara and Mediterranean region. According to analysis results, nettles phenolic compounds and contents were different between regions and parts of nettles. The difference between parts of nettle shows that all the parts of nettle could have different usages. In comparison of nettle tea and dried nettle from other researches, fresh samples phenolic contents were higher than others. For that reason, fresh nettle consumption could be healthier.

References

A. K. Ayan, O. Caliskan, and C. Cirak, “Isırgan Otu (Urtica spp.) Ekonomik Önemi ve Tarımı,” OMÜ Agricultural Faculty, vol. 21, no. 3, pp. 357–363, 2006.

K. Taylor, “Biological flora of the British Isles: Urtica dioica L,” Journal of Ecology, vol. 97, no. 6, pp. 1436–1458, 2009.

S. Hosbas, Urtica dioica L. Bitkisi Üzerinde Farmakognozik Araştırmalar [M.S. thesis], T.C. Gazi University, Institute of Medical Sciences Pharmacognosy Department, 2008.

N. Sekeroglu, F. Ozkutlu, M. Deveci, O. Dede, and N. ve Yılmaz, “Ordu ve Yöresinde Sebze Olarak Tüketilen Bazı Yabani Bitkilerin Besin Değeri Yönünden İncelenmesi,” in Turkey VI. Arable Crops Congress, pp. 523–528, Antalya, Turkey, September 2005.

P. M. Guarrera, “Traditional phytotherapy in Central Italy (Marche, Abruzzo, and Latium),” Fitoterapia, vol. 76, no. 1, pp. 1–25, 2005.

A. Ziyyat, A. Legssyer, H. Mekhfi, A. Dassouli, M. Serhrouchni, and W. Benjelloun, “Phytotherapy of hypertension and diabetes in oriental Morocco,” Journal of Ethnopharmacology, vol. 58, no. 1, pp. 45–54, 1997.

Y. C. Fiamegos, C. G. Nanos, J. Vervoort, and C. D. Stalikas, “Analytical procedure for the in-vial derivatization—extraction of phenolic acids and flavonoids in methanolic and aqueous plant extracts followed by gas chromatography with mass-selective detection,” Journal of Chromatography A, vol. 1041, no. 1-2, pp. 11–18, 2004.

C. Proestos, I. S. Boziaris, G. J. E. Nychas, and M. Komaitis, “Analysis of flavonoids and phenolic acids in Greek aromatic plants: investigation of their antioxidant capacity and antimicrobial activity,” Food Chemistry, vol. 95, no. 4, pp. 664–671, 2006.

R. Kraus and G. Spiteller, “Phenolic compounds from roots of Urtica dioica,” Phytochemistry, vol. 29, no. 5, pp. 1653–1659, 1990.

T. Kraushofer and G. Sontag, “Determination of some phenolic compounds in flax seed and nettle roots by HPLC with coulometric electrode array detection,” European Food Research and Technology, vol. 215, no. 6, pp. 529–533, 2002.

N. Chaurasia and M. Wichtl, “Flavonolglykoside aus Urtica dioica,” Planta Medica, vol. 53, no. 5, pp. 432–434, 1987.

N. S. Kavtaradze and M. D. Alaniya, “Anthocyan glucosides from Urtica dioica,” Chemistry of Natural Compounds, vol. 39, no. 3, p. 315, 2003.

T. F. Ji, C. H. Liu, A. G. Wang et al., “Studies on the chemical constituents of Urtica dioica L. grown in Tibet Autonomous Region,” Zhong Yao Cai, vol. 30, no. 6, pp. 662–664, 2007.

H. Sovová, M. Sajfrtová, M. Bártlová, and L. Opletal, “Near-critical extraction of pigments and oleoresin from stinging nettle leaves,” Journal of Supercritical Fluids, vol. 30, no. 2, pp. 213–224, 2004.

M. Hojnik, M. Škerget, and Ž. Knez, “Isolation of chlorophylls from stinging nettle (Urtica dioica L.),” Separation and Purification Technology, vol. 57, no. 1, pp. 37–46, 2007.

J. L. Guil-Guerrero, M. M. Rebolloso-Fuentes, and M. E. Torija Isasa, “Fatty acids and carotenoids from Stinging Nettle (Urtica dioica L.),” Journal of Food Composition and Analysis, vol. 16, no. 2, pp. 111–119, 2003.

R. Petlevski, M. Hadžija, M. Slijepčevič, and D. Juretič, “Effect of “antidiabetis” herbal preparation on serum glucose and fructosamine in NOD mice,” Journal of Ethnopharmacology, vol. 75, no. 2-3, pp. 181–184, 2001.

I. Gülçin, Ö. I. Küfrevioǧlu, M. Oktay, and M. E. Büyükokuroǧlu, “Antioxidant, antimicrobial, antiulcer and analgesic activities of nettle (Urtica dioica L.),” Journal of Ethnopharmacology, vol. 90, no. 2-3, pp. 205–215, 2004.

N. Gul, S. A. Ahmed, and L. A. Smith, “Inhibition of the protease activity of the light chain of type A botulinum neurotoxin by aqueous extract from stinging nettle (Urtica dioica) leaf,” Basic and Clinical Pharmacology and Toxicology, vol. 95, no. 5, pp. 215–219, 2004.

Y. S. Velioglu, G. Mazza, L. Gao, and B. D. Oomah, “Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products,” Journal of Agricultural and Food Chemistry, vol. 46, no. 10, pp. 4113–4117, 1998.

M. P. Kähkönen, A. I. Hopia, H. J. Vuorela et al., “Antioxidant activity of plant extracts containing phenolic compounds,” Journal of Agricultural and Food Chemistry, vol. 47, no. 10, pp. 3954–3962, 1999.

Anonymous, 2011, http://www.kimyaevi.org

V. L. Singleton and J. A. Rossi, “Colorimetry of total phenolics with phosphomolybdic-phosphotungstic asit reagents,” American Journal of Enology and Viticulture, vol. 16, pp. 144–158, 1965.

V. L. Singleton, R. Orthofer, and R. M. Lamuela-Raventós, “Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent,” Methods in Enzymology, vol. 299, pp. 152–178, 1998.

W. Brand-Williams, M. E. Cuvelier, and C. Berset, “Use of a free radical method to evaluate antioxidant activity,” LWT—Food Science and Technology, vol. 28, no. 1, pp. 25–30, 1995.

T. Akis, Piyasada Çay Olarak Tüketilen Bazı Bitkilerin Belirlenmesi ve Fenolik Yapılarının İncelenmesi, M.S. thesis, Ege University, Institute of Science, 2010.

J. Hudec, M. Burdová, L. Kobida et al., “Antioxidant capacity changes and phenolic profile of Echinacea purpurea, nettle (Urtica dioica L.), and dandelion (Taraxacum officinale) after application of polyamine and phenolic biosynthesis regulators,” Journal of Agricultural and Food Chemistry, vol. 55, no. 14, pp. 5689–5696, 2007.

P. Pinelli, F. Ieri, P. Vignolini, L. Bacci, S. Baronti, and A. Romani, “Extraction and HPLC analysis of phenolic compounds in leaves, stalks, and textile fibers of Urtica dioica L,” Journal of Agricultural and Food Chemistry, vol. 56, no. 19, pp. 9127–9132, 2008.

C. Ozcan, Semizotu, Isırgan Otu, Menengiç ve Kuşburnu Gibi Tıbbi ve Aromatik Bitkilerde Flavonollerin HPLC-MS ile Tayini, Ph.D. thesis, Fırat University, Institute of Science, 2009.