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Biological Diversity and Conservation
ISSN 1308-8084 Online
ISSN 1308-5301 Print
Research article/Araştırma makalesi
DOI: 10.46309/biodicon.2021.881775
14/1 (2021) 93-97
Heavy metal content screening in leaves and flowers of Hypericum origanifolium by atomic absorption
spectrometry
Asiye BERBER 1, İsmühan POTOĞLU ERKARA *2, Murat OLGUN 2, Onur KOYUNCU 2, Murat ARDIÇ 2,
Okan SEZER 2
ORCID: 0000-0002-8340-4793; 0000-0001-5780-4999; 0000-0002-6234-455X; 0000-0002-0364-6638;
0000-0001-8734-3038; 0000-0001-7304-1346
1
Eskişehir Osmangazi Univ, Faculty of Education, Department of Elementary Science Education, Eskişehir, Turkey
2
Eskişehir Osmangazi University, Faculty of Science and Letters, Department of Biology, Eskişehir, Turkey
3
Eskişehir Osmangazi University, Agricultural Faculty, Field Crop Department, Eskişehir, Turkey
Abstract
Atomic absorption spectrometry facilitates the reliable determination of mineral content during pharmaceutical
quality control of medicinal plants. In the present work, measurable amounts of Fe, Ca, Cu, K, Mg, Mn, Na and Zn
were detected in the leaves and flowers of Hypericum origanifolium Willd. through atomic absorption spectrometry.
Mean heavy metal content in the flowers and leaves of H. origanifolium was, in descending order, Ca > Mg > K > Na >
Fe. Ca was present in higher concentrations in the flowers (11157.24 ppm) and leaves (20132.24 ppm) of titled plant.
Our results reveal that flowers are less suitable as target plant parts for metal accumulation than leaves.
Keywords: Hypericum origanifolium, heavy metals, atomic absorption spectrometry
---------- ---------Hypericum origanifolium’un yaprak ve çiçeklerindeki ağır metal içeriğinin atomik absorpsiyon spektrometresi
ile belirlenmesi
Özet
Atomik absorpsiyon spektrometresi, tıbbi bitkilerin farmasötik kalite kontrolü sırasında mineral içeriğinin
güvenilir bir şekilde belirlenmesini kolaylaştırmaktadır. Bu çalışmada, Hypericum origanifolium Willd'un yaprak ve
çiçeklerinde atomik absorpsiyon spektrometresi ile ölçülebilir miktarda Fe, Ca, Cu, K, Mg, Mn, Na ve Zn bulunduğu
tespit edilmiştir. H. origanifolium'un yaprak ve çiçeklerindeki ortalama ağır metal içeriği sırasıyla Ca> Mg> K> Na> Fe
olarak belirlenmiştir. Kalsiyum, çiçek (11157,24 ppm) ve yapraklarda (20132,24 ppm) en yüksek konsantrasyonda
tespit edilmiştir. Elde edilen sonuçlar çiçeklerin yapraklara kıyasla çok daha düşük seviyede ağır metal birikimi
gösterdiğini ortaya koymaktadır.
Anahtar kelimeler: Hypericum origanifolium, ağır metaller, atomik absorpsiyon spektrometresi
1.
Introduction
Especially in developing countries, environmental pollutants which are released to nature in an uncontrolled
manner reach the structure of plants through water, soil and air. Metals have an important place among these pollutants.
The use of metal based pollutant exposed plants by humans causes serious health problems such as kidney damage,
renal failure and liver damage [1; 2; 3]. Iron, zinc, calcium, magnesium, copper, potassium, manganese, sodium were
chosen as representative metals whose levels in the environment represent a reliable index of environmental pollution
and human health. Some plants like as Hypericum perforatum, Avena sterilis, Bifora radians, Chenopodium album,
Corresponding author / Haberleşmeden sorumlu yazar: Tel.: +905326338464; Fax.: +905326338464; E-mail: ismuhan@ogu.edu.tr
© Copyright 2021 by Biological Diversity and Conservation
Received: 16.02.2021;
Published: 15.04.2021
BioDiCon. 952-0221
*
94
Biological Diversity and Conservation – 14 / 1 (2021)
Consolida regalis, Humulus lupulus, Reseda lutea, Solanum nigrum, Sorghum halepense and Xanthium strumarium can
then be used as biomonitors for the determination of trace element levels [4; 5; 6; 7].
Hypericum L., which is a member of the Hypericaceae (Guttiferae) family, is represented approximately 500
species all around to world. About 108 of these species are naturally distributed in Turkey [8; 9; 10; 11]. There has been
many studies previously conducted on the Hypericum species, but very limited studies for H. origanifolium. According
to the literature reviews, only one anatomical study on H. origanifolium was found [12]. One of the most important
features of Hypericum taxa is their unique secondary metabolites. Hypericum taxa and their metabolites are widely used
in traditional and modern medicine today [13]. In the light of phytochemical studies performed on H. origanum,
flavonoids (myrcetin, rutin, quercetin, hyperoside), xanthones (iso-magniferin and mangiferin), naphthodiantrones
(frangulin, emodin, proto-pseudohypericin, psudohypericin and phenolicin) and phenolic acids have been determined at
the aerial parts of the plant [14; 15; 16; 17; 18; 19; 20].
Data available on the biological activity of H. origanifolium is also limited to a few previous reports that have
demonstrated their cytotoxic, antiproliferative, antimicrobial, antibacterial, antiyeast and antioxidant activities [18; 19;
20; 21; 22; 23; 24; 25; 26]. In the light of literature data, any studies on the heavy metal content of H. origanifolium not
determined.
The main purpose of this study is to determine the heavy metal content in extracts obtained from the flowers
and leaves of H. origanifolium by atomic absorption spectrometry. We hope that the relationship between the heavy
metal amounts in H. origanifolium leaves and flowers will be reference for future studies.
2. Materials and methods
H. origanifolium Willd. was collected around of Eskişehir, Turkey. H. origanifolium Willd. Sivrihisar, Tekören
village, 1100 m. June 2003 (OUFE 10334). The plant was identified according to Flora of Turkey and the East Aegan
Islands [8; 27; 28].
For the solid samples with a nitric acid (Merck, Darmstadt, Germany)–perchloric acid (Merck, Darmstadt,
Germany) digestion was used for mineralizing. The dried flowers and leaves of H. origanifolium were extracted for the
solution phase as described previously and analyzed for Fe, Zn, Ca, Mg, Cu, K, Mn, Na (Merck AAS standard
solutions), using Hitachi (180-70) Polorized Zeeman flame atomic absorption spectrometry [29; 30]. All precautions
were taken to prevent metal contamination, i.e. samples were cleaned with 2% HNO 3, rinsed in distilled water and
baked at 600°C. All samples were analyzed in triplicate and the mean values were calculated. In order to increase the
reliability of the measurements during the study, the instrument was calibrated at every 10 readings.
The flame atomic absorption spectrometry (FAAS) instrumental and operating conditions that provided the
best sensitivity for the determination of metal content are detailed in Table 2.1.
Table 2.1. FAAS instrumental parameters employed to determine metals
Burner
Wavelength
Slit width
Elements
Flame type
height (mm)
(nm)
(nm)
Fe
Air-C2H2
7.5
248.3
0.2
Zn
Air-C2H2
7.5
213.8
1.3
Ca
Air-C2H2
12.5
422.7
2.6
Mg
Air-C2H2
7.5
285.2
2.6
Cu
Air-C2H2
7.5
324.8
1.3
K
Air-C2H2
7.5
766.5
2.6
Mn
Air-C2H2
7.5
279.5
0.4
Na
Air-C2H2
7.5
589.0
0.4
Lamp
Current (mA)
10
10
7.5
7.5
7.5
10
7.5
10
Fuel gas
(1 min-1)
2.3
2.0
2.6
1.6
2.3
2.3
2.3
2.2
3. Results
In this study, the heavy metal contents of the leaves and flowers of Hypericum origanifolium Willd. were
investigated. These levels were obtained through flame atomic absorption spectrometry. Fe, Zn, Ca, Mg, Cu, K, Mn and
Na were determined to be present in the samples. Metal concentrations in the leaves of H. origanifolium were found to
be 130.25, 30.99, 20132.24, 7775.84, 7.60, 3361.53, 48.40, and 145.02 ppm; and in flower of H. origanifolium were
found to be 93.64, 26.26, 11157.23, 5768.23, 11.37, 2454.99, 36.12, and 184.94 ppm for iron, zinc, calcium,
magnesium, copper, potassium, manganese, sodium, respectively (Table 3.1. and Figure 3.1.).
Heavy Metal Content Screening in Leaves and Flowers of Hypericum origanifolium Willd. by Atomic Absorption Spectrometry
Asiye BERBER, İsmühan POTOĞLU ERKARA, Murat OLGUN, Onur KOYUNCU, Murat ARDIÇ, Okan SEZER
Biological Diversity and Conservation – 14 / 1 (2021)
95
Table 3.1. Element concentrations of H. origanifolium (ppm)
Elements
H. origanifolium (leaf)
H. origanifolium (flower)
Fe
130.25
93.64
Zn
30.99
26.26
Ca
20132.24
11157.23
Mg
7775.84
5768.23
Cu
7.60
11.37
K
3361.53
2454.99
Mn
48.40
36.12
Na
145.02
184.94
Figure 3.1. Relationship between leaf and flower metal concentrations of H. origanifolium
4. Conclusions and discussion
In the light of obtained data from flame atomic absorption spectrometry analysis of the leaves and flowers of
Hypericum origanifolium, Fe, Zn, Ca, Mg, K and Mn concentrations were recorded as higher in the leaf than in the
flower, 130.25 > 93.64 ppm, 30.99 > 26.26 ppm, 20132.24> 11157.23 ppm, 7775.84> 5768.23 ppm. 3361.53> 2454.99
ppm, 48.40> 36.12 ppm, respectively. Cu and Na concentrations were observed to be higher in the flower than in the
leaf 11.37> 7.60 ppm, 184.94 > 145.02 ppm, respectively (Figure 1, Table 2).
The mean heavy metal content in the flowers and leaves of H. origanifolium was, in descending order, Ca >
Mg > K > Na > Fe. Ca was present in higher concentrations in the flowers (11157.24 ppm) and leaves (20132.24 ppm)
of the titled plant. Gomez et al. (2004) indicated that the CA concentration in H. perforatum was 100-500 ppm for the
dried herb, as indicated in table 3.1. [4]. This situation shows that the Ca concentration of the leaf and flower of H.
origanifolium is higher than H. perforatum. Kadıoğlu et al. found the concentrations to be 495, 62.6, 11.1, 19,5 ppm for
iron, manganese, copper, zinc, respectively [5]. In this study, the concentrations of Fe and Mn were higher, while that of
Zn was lower than in the H. perforatum concentrations reported by Kadıoğlu et al. (2005).
Findings obtained from elemental analysis studies on plants reveal that herbal foods are rich in especially Mg,
Ca, Na and K [3]. The data obtained in our study coincide with the above findings. K, Mg and Ca which are represent
the most abundant metal constituents of many plants were identified as the most abundant metals in H. origanifolium
extracts. Internal and external factors have direct and indirect effects on the different metal concentrations of distinct
parts of plants. These regional differences in metal content can occur due to many different reasons, such as genetic
factors, growth conditions, analytical procedures and geographical variations. These factors are directly affected by the
location differences where the plant samples are collected. Hence, there is an indirect relationship between sample
locations and metal content can be mentioned. Though much is known about the functional role of a number of
elements, the best foreseeable benefit for human health, mineral nutrition, lies in obtaining the correct amount of
supplementation in the right form at the right time. High or low levels of Ca, Cu, Zn, Mn, K and Mg may invite many
disorders [3]. These elements also play a part in neurochemical transmission, as well as serving as constituents of
biological molecules, as a cofactor for various enzymes such as NAD(P)H oxidase, Ferroxidase, Alcohol
dehydrogenase and in a variety of different metabolic processes.
Heavy Metal Content Screening in Leaves and Flowers of Hypericum origanifolium Willd. by Atomic Absorption Spectrometry
Asiye BERBER, İsmühan POTOĞLU ERKARA, Murat OLGUN, Onur KOYUNCU, Murat ARDIÇ, Okan SEZER
96
Biological Diversity and Conservation – 14 / 1 (2021)
High amounts of Ca are expected one way or another, as it is one of the most common minerals of the soil,
from where it is readily absorbed into the plants. Iron is one of the important elements for the human body and is
involved in vital activities such as hemoglobin formation, oxygen and electron transfer. Zinc and Copper, on the other
hand, are involved in many different vital processes such as being important components of enzymatic and redox
systems. For this reason, they have an important place among the elements that should be found regularly in the diet [6].
Also, the consequences show that most of these herbal plants contain vital elements for human metabolism and are also
essential for growth and development as well as for the prevention and cure of diseases.
While many investigations into the quality values of medicinal plants are being reported in the current
literature [7], less emphasis has been made on the metal content of herbal products. Metallic elements are constituent
plant compounds demonstrating biological activity as essential or toxic agents in metabolism. Thus, the application of
metal monitoring as a pattern recognition method in medicinal herbs is a promising tool for their characterization.
Chemotaxonomy is one of the most important tools that help classify plant taxa. The field of study of
chemotaxonomy is to reveal the chemical content similarities and differences between different taxonomic groups. In
this context, determining the metal content of plants is extremely valuable for chemotaxonomic researches. Heavy metal
levels are important pollutants for soil, water, plant, the environment and human health. Especially the accumulation of
heavy metals in some plant taxa makes them a valuable tool in determining heavy metal pollution. Therefore, further
investigations are also needed to determine interactions between the leaves and flowers in terms of heavy metals.
Acknowledgements
This study was supported by Anatolia Science Academy in scope of the scientific project named as
“Eskişehirde Doğal Yayılış Gösteren Bazı Hypericum L. Taksonlarının Vejetatif ve Generatif Organlarındaki
Ağır Metal İçeriğinin Atomik Absorpsiyon Spektrofotometresi ile Belirlenmesi (in Turkish)” .
References
[1]
[2]
[3]
[4]
[5]
[6]
[7]
[8]
[9]
[10]
[11]
[12]
[13]
[14]
[15]
[Shaw, D., Leon, C., Kolev, S. & Murray, V. (1997). Traditional remedies and food supplements. A 5-year
toxicological study (1991-1995). Drug Safety, 17, 342-356.
Andrew, A. S., Warren, A. J., Barchowsky, A., Temple, K. A., Klei, L., Soucy, N. V., O’Hara, K. A. &
Hamilton, J. W. (2003). Genomic and Proteomic Profiling of Responses to Toxic Metals in Human Lung
Cells. Environ. Health Perspect, 111(6), 825-835.
Ajasa, A. M. O., Bello, M. O., İbrahim, A. O., Ogunwande, I. A. & Olawore, N. O. (2004). Heavy trace
metals and macronutrients status in herbal plants of Nigeria. Food Chemistry, 85, 67-71.
Gomez, M. R., Cerutti, S., Olsina, R. A., Silva, M. F. & Martinez, L. D. (2004). Metal content monitoring in
Hypericum perforatum pharmaceutical derivatives by atomic absorption and emission spectrometry. Journal
of Pharmaceutical and Biomedical Analysis, 34(3), 569-576.
Kadıoğlu, I., Mendil, D., Sarı, H. & Hasdemir, E. (2005). Determination of Heavy Metal Levels in Some
Weeds Collected from Tokat, Turkey. Asian Journal of Chemistry, 17(1), 564-568.
Özcan, M. M. & Akbulut, M. (2007). Estimation of minerals, nitrate and nitrite contents of medicinal and
aromatic plants used as spices, condiments and herbal tea. Food Chemistry, 106, 852-858.
Gomez, M. R., Cerutti, S., Sombra, L. L., Silva, M. F. & Martinez, L. D. (2007). Determination of heavy
metals for the quality control in argentinian herbal medicines by ETAAS and ICP-OES. Food and Chemical
Toxicology, 45, 1060-1064.
Robson, N. K. B. (1967). Hypericum L. – In Flora of Turkey and the East Aegan Islands (Ed. Davis PH) Vol.
2. Edinburgh University Press, p. 355.
Robson, N. K. B. (1977). Studies in the genus Hypericum L. (Guttiferae): 1. Infrageneric classification.
Bulletin of the British Museum (Natural History) Botany, 5: 291-355.
Robson, N. K. B. (2012). Studies in the genus Hypericum L. (Hypericaceae) 9. Addenda, corrigenda, keys,
lists and general discussion. Phytotaxa 72, 1-111.
Duman, H. & Çakır-Dindar, E. G. (2020). Hypericum alacamdaglariense (Hypericaceae), a new species from
Turkey. Phytotaxa 470(2), 176-185.
Potoğlu Erkara, İ. & Tokur, S. (2004). Morphological and Anatomical investigations on some Hypericum L.,
species growing naturally in and arround Eskişehir. Trakya Univ J Sci 5, 97-105.
Yaylacı, Ö. K., Özgişi, K., Sezer, O., Orhanoğlu, G., Öztürk, D. & Koyuncu, O. (2013). Anatomical studies
and conservation status of rare endemic Hypericum sechmenii Ocak & Koyuncu (Sect: Adenosepalum) from
Eskişehir-Turkey. Journal of Selcuk University Natural and Applied Science, 2(1), 1-11.
Mathis, C. & Ourisson, G. (1964). Chemo-taxonomic study of the genus Hypericum. III. The distribution of
saturated hydrocarbons and monoterpenes from the essential oil of Hypericum. Phytochem, 3, 133-141.
Kitanov, G. M. & Nedialkov, P. T. (1998). Mangiferin and isomangiferin in some Hypericum species.
Biochem Syst Ecol, 26, 647-653.
Heavy Metal Content Screening in Leaves and Flowers of Hypericum origanifolium Willd. by Atomic Absorption Spectrometry
Asiye BERBER, İsmühan POTOĞLU ERKARA, Murat OLGUN, Onur KOYUNCU, Murat ARDIÇ, Okan SEZER
Biological Diversity and Conservation – 14 / 1 (2021)
[16]
[17]
[18]
[19]
[20]
[21]
[22]
[23]
[24]
[25]
[26]
[27]
[28]
[29]
[30]
97
Makovests'ka, O. Y. (1999). Research of Biologically Active Substances of Hypericum L. species. Report VI.
Sections olympia (Spach) Nyman, oampylopus Boiss. and origanifolia Stef. Farm Zh (Kiev), 6, 46-50.
Sirvent, T. M., Walker, L., Vance, N. & Gibson, D. M. (2002). Variation in hypericins from wild populations
of Hypericum perforatum L. in the Pacific Northwest of the USA. Econ Bot, 56, 41-48.
Çırak, C., Radušienė, J., Ivanauskas, L. & Janulis, V. (2007). Variation of bioactive secondary metabolites in
Hypericum origanifolium during its phenological cycle. Acta Physiologiae Plantarum, 29(3), 197-203.
Öztürk, N., Tunçel, M. & Potoğlu Erkara, İ. (2009). Phenolic compounds and antioxidant activities of some
Hypericum species: A comparative study with H. perforatum. Pharmaceutical biology, 47(2), 120-127.
Bertoli, A., Çırak, C. & Seyis, F. (2015). Hypericum origanifolium Willd.: The essential oil composition of a
new valuable species. Industrial Crops and Products, 77, 676-679.
Sakar, M. K., Tamer, A. U. & Tokur, S. (1988). Antimicrobial activities of some Hypericum species growing
in Turkey. Fitoterapia, 59, 49-52.
Sakar, M. K. & Tamer, A. U. (1990). Antimicrobial activity of different extracts from some Hypericum
species. Fitoterapia, 61, 464-466.
Güzey. G. (2007). Cytotoxic and Antiproliferative Effects of Hypericum perforatum, Hypericum montbretti
and Hypericum origanifolium Species on A549, HeLa and NIH3T3 Cell Cultures. PhD Thesis, Anadolu
University Institute of Health Sciences, Eskişehir, Turkey.
Güzey, G., Ibadova, S., Öztürk, Y., Öztürk, N., Maggi, F., Sagratini, G., Ricciutelli, M. & Vittori, S. (2011).
Antiproliferative and Antioxidant Effects of Three Hypericum Species of Turkish Origin: H. perforatum, H.
montbretii and H. origanifolium. Medicinal and Aromatic Plant Science and Biotechnology, 5(1), 91-99.
Yaşar, Ş. N., Can, Ö. D., Öztürk, N., Sagratini, G., Ricciutelli, M., Vittori, S. & Maggi, F. (2013). Central
nervous system activities of Hypericum origanifolium extract via GABAergic and opioidergic mechanisms.
Phytotherapy Research, 27(6), 877-884.
Boran, R. (2018). Investigations of anti-aging potential of Hypericum origanifolium Willd. for skincare
formulations. Industrial Crops and Products, 118, 290-295.
Robson, N. K. B. (1988). Hypericum L. – In Flora of Turkey and the East Aegan Islands (Eds. Davis PH,
Mill RR and Tan K) Vol.10. Edinburgh University Press, p. 96.
Dönmez, A. A. (2000). Hypericum L. – In Flora of Turkey and the East Aegan Islands (Eds. Güner A,
Özhatay N, Ekim T and Başer KHC) Vol.11. Edinburgh University Press, pp. 71-72.
Que Hee, S. S. & Boyle, Jr. (1988). Simultaneous Multielemental Analysis of Some Environmental And
Biological Samples By Inductively Coupled Plasma Atomic Emission-Spectrometry. Analytical Chemistry,
60(10), 1033-1042.
Kılıç, M., Ay, G., Koçbaş, F., Kılıç, F.M. (2019). Determination level of heavy metal in Ayvalik Saltern
using Halimione portulacoides (L.) plant. Biological Diversity and Conservation, 12(1), 100-106.
Heavy Metal Content Screening in Leaves and Flowers of Hypericum origanifolium Willd. by Atomic Absorption Spectrometry
Asiye BERBER, İsmühan POTOĞLU ERKARA, Murat OLGUN, Onur KOYUNCU, Murat ARDIÇ, Okan SEZER