The Determination of Mineral and Heavy Metal Contents of Echinacea Species Cultivated in Turkey

This study was carried out in the ecological conditions of Konya in 2013 and 2014 at Selcuk University Faculty of Agriculture, The Farm of Medicinal Plants. Mineral compositions (Na, K, Ca, P, Fe, Zn, Cu, Mn Al, Cd, Co, Ni, Cr, Mo and Pb) were examined in five different species of Echinacea. The analyses of mineral compositions in Echinacea species were determined using NMKL 161 method and A Perkin-Elmer Optima 2000 inductively coupled plasma–optical emission spectrophotometer (ICP–OES) was used to analyze the elements to determine the mineral substances. The highest B, Ca, K, Na, P and S average contents according to the two-year mean values belonging to the herbs of Echinacea species were Echinacea purpurea var. double decker 135,029 ppm, Echinacea purpurea var. double decker 31185,018 ppm, Echinacea purpurea var. baby white swan 21968,858 ppm, Echinacea pallida var. pallida 104,742 ppm, Echinacea purpurea var. purpurea 2490,672 ppm and Echinacea paradoxa var. paradoxa 1754,594 ppm respectively. The herbs with the highest Al, Cd, Co, Cr, Cu, Fe, Mn, Pb, Mo, Ni and Zn average contents according to the two-year mean values belonging to the herbs of Echinacea species were Echinacea purpurea var. baby white swan 432.195 ppm, Echinacea purpurea var. purpurea 0.111 ppm, Echinacea paradoxa var. paradoxa 0,395 ppm, Echinacea paradoxa var. paradoxa 2,615 ppm, Echinacea purpurea var. purpurea 22,535 ppm, Echinacea purpurea var. baby white swan 451.350 ppm, Echinacea paradoxa var. paradoxa 46.217 ppm, Echinacea paradoxa var. paradoxa 21.366 ppm, Echinacea purpurea var. purpurea 3.364 ppm, Echinacea paradoxa var. paradoxa 2.936 ppm and Echinacea paradoxa var. paradoxa 22.218 ppm respectively. The highest B, Ca, K, Na, P and S contents according to the two-year mean values of Echinacea roots were determined as Echinacea purpurea var. double decker 102,620 ppm, Echinacea purpurea var. purpurea 38805,7 ppm, Echinacea paradoxa var. paradoxa 20656,313 ppm, Echinacea pallida var. pallida 2062,226 ppm, Echinacea paradoxa var. paradoxa 2262,165 ppm and Echinacea paradoxa var. paradoxa 2181,838 ppm.


INTRODUCTION
Compositeae (Asteraceae) is the richest family of flowering plants, representing by nearly 1000 genera and close to 20,000 species.About 133 genera and 1156 species belonging to family Asteraceae are grown in our country [1].Echinacea species are perennial herbaceous plants in the Asteraceae family [2].The Echinacea genus contains 11 species according to the review by Mc Keown (1999) [4].The first culture studies of Echinacea plant in our country started in 2005 in the Selcuk University Faculty of Agriculture, The Farm of Medicinal Plants.Echinacea is a plant that can be grown up to 60-80 cm and starts to bloom from the second half of June.Leaf and stem of them are light hairy.The stem is cylindrical and consists of a very large number of side branches (25-30 on average).Approximately 250-300 seeds can be obtained in a mature flower bed.The seeds are about 5 mm long and 1.5 mm wide; it is angular and resembles a funnel.1000 seed weights are about 5-6 g [5].
Echinacea species were used as wound healing externally and as a headache, gastric pain and cough cutter by the Native Americans [3,6].It has proven anti-bacterial, anti-inflammatory, immune system strengthening and wound healing properties [7].Especially in infectious diseases, it helps the body naturally increase resistance and pain.It is a preventive and therapeutic against colds, chill and influenza.The demand to Echinacea is increasing day by day both in our country and in the world [8].Echinacea species are imported as drug or preparate and used in different pharmaceutical forms in the world.
This study was carried out in the ecological conditions of Konya in 2013 and 2014 at Selcuk University Faculty of Agriculture, The Farm of Medicinal Plants.Mineral compositions (macro elements, Na, K, Ca, and P; micro elements, Fe, Zn, Cu, and Mn; heavy metals, Al, Cd, Co, Ni, Cr, Mo and Pb) were investigated in herb and root specimens obtained from during full bloom period of five different species of Echinacea ((Echinacea purpurea var.purpurea (L.) Moench, Echinacea pallida var.pallida (Nutt.)Nutt., Echinacea paradoxa var.paradoxa (J.B.S. Norton) Britton, Echinacea purpurea var.baby white swan (L.) Moench and Echinacea purpurea var.double decker (L.) Moench)).

Plant Material
This study was carried out in the ecological conditions of Konya in 2013 and 2014 at Selcuk University Faculty of Agriculture, The Farm of Medicinal Plants.The seedlings of Echinacea species were growed in the greenhouse in 2012.Because echinacea is a perennial plant, analyses samples were not obtain from the farm the first year.But harvested analyses samples in the 2nd and 3rd years (2013)(2014).The herb samples of five different species of Echinacea (Echinacea purpurea var.purpurea (L.) Moench, Echinacea pallida var.pallida (Nutt.)Nutt., Echinacea paradoxa var.paradoxa (J.B.S. Norton) Britton, Echinacea purpurea var.baby white swan (L.) Moench and Echinacea purpurea var.double decker (L.) Moench)) harvested during the full blooming period were dried in the shade and ground for analysis.During the short storage time, the plant samples were kept at -20°C until use.

Determination of Mineral and Heavy Metal Contents of Echinacea Species Cultivated in Turkey
The NMKL 161 method was used to determine the mineral substances.For this purpose, ICP-OES (A Perkin-Elmer Optima 2000 inductively coupled plasma-optical emission spectrophotometer) was used.The determined minerals were Al, Co, Mo, Ca, B, Cd, Cr, Cu, Fe, K, Mn, Na, Ni, P, Pb, S and Zn.The results of the mineral substances were given in ppm (mg / kg) [9].

Evaluation of Data
Variance analyzes of the data obtained in the study were made by JMP-SAS 7.0 statistical package program and MSTAT-C statistical package program was used to make the letters.The statistically significant group tests between the mean values were determined by LSD (0.01 and 0.05 significance levels).

Minerals and Heavy Metals Results of Herbs
Mineral compositions (macro elements, Na, K, Ca, and P; micro elements, Fe, Zn, Cu, and Mn; heavy metals, Al, Cd, Co, Ni, Cr, Mo and Pb) were determined five different species of Echinacea ((Echinacea purpurea var.purpurea (L.) Moench, Echinacea pallida var.pallida (Nutt.)Nutt., Echinacea paradoxa var.paradoxa (J.B.S. Norton) Britton, Echinacea purpurea var.baby white swan (L.) Moench and Echinacea purpurea var.double decker (L.) Moench)) The average values of mineral substances in the herbs obtained during the full blooming period in 2013 and 2014 are given in the tables section..While the highest B (Boron) amount, according to two year mean values, was found in Echinacea purpurea var.double decker with 135,029 ppm, the lowest was found in Echinacea paradoxa var.paradoxa with 58,317 ppm.According to years, the mean values of 2014 was found to be higher with 120,306 ppm with respect to the B amount.Although the highest Ca (Calcium) value of 2013 and 2014 mean values was found in 2014 with 28182.381ppm, it took place in the same group with the average of 2013 according to the statistical evaluation.According to two-year mean values of herbs, the highest Ca amount was found in Echinacea purpurea var.double decker with 31185.018ppm.When two-year mean values were analyzed, the highest K (potassium) concentration was found in Echinacea purpurea var.baby white swan with 21968.858ppm and the lowest was found in Echinacea paradoxa var.paradoxa with 18922.968ppm.When 2013 and 2014 year mean values were analyzed, the highest K value was found to be higher as 20957.844ppm in 2014.When the two-year mean values of the herbs were examined, the highest Na (sodium) concentration was found in Echinacea pallida var.pallida with 104,742 ppm and the lowest Na concentration was found in Echinacea purpurea var.baby white swan with 41,605 ppm.According to the data of 2013 and 2014, Na in 2013 was found to be 84.238ppm higher than in 2014.The highest P (phosphor) intensity according to the two-year mean values was found as 2490,672 ppm in E. purpurea var.purpurea.Although the average P values of 2013 and 2014 were statistically in the same group, the average value for 2014 was higher than 2013 with 2271.622ppm.The highest value of the herbs were determined to be 1754.594ppm in Echinacea paradoxa var.paradoxa according to the mean values of S (sulphur) content for both year.Also, the lowest was found in Echinacea purpurea var.purpurea with 1087.522ppm.When both years mean values were considered, S value of 2013 was found to be higher with 1437.124ppm.
Results belonging to Al, Cd, Co, Cr, Cu, Fe, Mn, Pb, Mo, Ni and Zn heavy metals are shown in "Tables" section.According to the average of the two-year values, the Al (aluminium) content intensity was found to be higher in 2013 with 324.281 ppm.Among the herbs, the highest Al amount was found in Echinacea purpurea var.baby white swan with 432.195 ppm.The lowest Al amount was found in Echinacea pallida var.pallida with 115,976 ppm.Looking at 2013 and 2014, while the Cd (cadmium) concentration was found to be higher in 2013 with 0.112 ppm, it was found to be lower in 2014 with 0.101 ppm.When two-year mean values of herbs were examined, the highest Cd amount was found in Echinacea purpurea var.purpurea with 0.111 ppm.According to the two-year mean values of the herbs, while the highest Co (cobalt) amount was found in Echinacea paradoxa var.paradoxa with 0,395 ppm, the lowest Co amount was found in Echinacea purpurea var.baby white swan with 0,316 ppm.When twoyear average values were considered the highest Cr (chromium) concentration was obtained in 2013 with 2,102 ppm.In the average of both two-year data of the herbs, the highest Cr concentration was obtained from Echinacea paradoxa var.paradoxa with 2.615 ppm and lowest in Echinacea pallida var.pallida with 0.544 ppm.Considering the mean values of 2013 and 2014, Cu (copper) was the highest with 17.095 ppm in 2014.When the average of the twoyear data of herbs was taken into consideration, the Cu measure was obtained from Echinacea purpurea var.purpurea with 22,535 ppm as the highest and Echinacea purpurea var.baby white swan with 11,581 ppm as the lowest.When the two-year mean values of the herbs were examined, the highest Fe (iron) amount was found in Echinacea purpurea var.baby white swan with 451,350 ppm and the lowest Fe amount was found in Echinacea paradoxa var.paradoxa with 158,181 ppm.When mean values of data taken between 2013 and 2014 were analyzed, Fe proportion was found to be higher with 369,942 ppm in 2013.Mn (manganese) proportion in 2013 was found higher than 2014 with 32.988 ppm.In the average of two-year data of the herbs, the highest Mn amount was determined in Echinacea paradoxa var.paradoxa with 46.217 ppm and the lowest in Echinacea pallida var.pallida with 9.126 ppm.When each year's Pb (Lead) mean values were examined, the average value in 2013 (0,966 ppm) was found higher than 2014.When the two-year mean values of the herbs were examined, it was seen that Echinacea paradoxa var.paradoxa had the highest Pb concentration with 1,366 ppm and Echinacea pallida var.pallida had the lowest value with 0,583 ppm.Mo (molybdenum) amount in 2013 was found higher than 2014 with 2.268 ppm.Looking at the mean values of 2013 and 2014, the highest Mo amount was determined in Echinacea purpurea var.purpurea with 3,364 ppm.The lowest Mo amount was found in Echinacea purpurea var.double decker with 0,673 ppm.In the average of two-year data of the herbs, the highest Ni (nickel) mass was determined in Echinacea paradoxa var.paradoxa with 2,936 ppm and the lowest in Echinacea pallida var.pallida with 0.941 ppm.Ni mass in 2013 was found higher than 2014 with 2.410 ppm.Zn (zinc) amount in 2013 was found higher than 2014 with 19.809 ppm.The highest Zn proportion was determined in Echinacea paradoxa var.paradoxa with 22.218 ppm according to two-year mean values of herbs.The lowest Zn concentration was found in Echinacea pallida var.pallida with 14,710 ppm.

Minerals and Heavy Metals Results of Roots
Mineral compositions (macro elements, Na, K, Ca, and P; micro elements, Fe, Zn, Cu, and Mn; heavy metals, Al, Cd, Co, Ni, Cr, Mo and Pb) were examined in root samples obtained during full bloom period of five different species of Echinacea (Echinacea purpurea var.purpurea (L.) Moench, Echinacea pallida var.pallida (Nutt.)Nutt., Echinacea paradoxa var.paradoxa (J.B.S. Norton) Britton, Echinacea purpurea var.baby white swan (L.) Moench and Echinacea purpurea var.double decker (L.) Moench) The average values of mineral substances in the roots obtained during the full blooming period in 2013 and 2014 are given in the "Tables" section.
When the two-year average B values of roots were examined, it was found to be the highest at Echinacea purpurea var.double decker root with 102.620 ppm.B content value was found to be higher in 2013 with 74.440 ppm according to the mean values at the roots in 2013 and 2014.According to years, Ca was found to be higher in 2013 with 37570.5 ppm, while it was found the highest in root of Echinacea purpurea var.purpurea with 38805.7 ppm when average two-year Ca concentration of roots was compared.When both years mean values were considered, K value in 2013 was found to be higher with 16853.219ppm.When the two-year average K amounts of plant roots were examined, Echinacea paradoxa var.paradoxa root was found the highest with 20656.313ppm.The lowest K mass was obtained from Echinacea purpurea var.baby white swan root with 10287.899ppm.When the average values of the two years were examined, the proportion of Na in 2014 with 379.402 ppm was higher than Na of 2013 (627.201ppm).According to the two-year mean Na values of the herbs' roots, the highest was examined in Echinacea pallida var.pallida root with 2062.226ppm and the lowest was found in Echinacea purpurea var.baby white swan with 605,483 ppm.When two-year mean values of P values were examined, the highest P concentration was found in Echinacea paradoxa var.paradoxa root with 2262.165ppm.P concentration, according to years, was found to be higher with 1856.940ppm in 2013.According to years, the average S amount in 2013 was found to be higher with 1988.549ppm.When the two-year average S amounts of plant roots were examined, it was found the highest in Echinacea paradoxa var.paradoxa root with 2181.838ppm.The lowest S amount was found in Echinacea purpurea var.purpurea root with 1441.389ppm.
When the data of heavy metals of Al, Cd, Co, Cr, Cu, Fe, Mn, Pb, Mo, Ni and Zn in roots were determined, according to the average of the two-year values, the Al content mass was found to be higher in 2013 with 2197.75 ppm.Among the roots, the highest Al mass was found in Echinacea purpurea var.purpurea with 3966.88 ppm.The lowest Al mass was found in Echinacea purpurea var.double decker with 415.25 ppm.Looking at the two-year average Cd proportions, 2013 was identified higher than 2014 with 0,144 ppm.When the two-year mean values of the roots were studied, the highest and the lowest Cd values were respectively; in Echinacea purpurea var.purpurea root with 0.174 ppm and in Echinacea purpurea var.purpurea root with 0.099 ppm.When two-year mean Co values of roots were studied, the highest was found in Echinacea purpurea var.purpurea root with 0,712191 ppm.Zn was found to be higher in 2013 with 0,496637 ppm according to the mean values at the roots in 2013 and 2014.Cr concentration, according to mean values of years, was found to be higher with 4,439 ppm in 2013.When the two-year mean values of the roots were determined, the highest Cr amount was found in Echinacea purpurea var.purpurea root as 7.755 ppm.When Cu mass of the roots were taken into account, the highest value was obtained from Echinacea purpurea var.purpurea root with 25.305 ppm.Cu amount was found to be higher with 24,548 ppm in 2014 among the mean values of 2013 and 2014.When the two-year average Fe mass of plant roots were considered, the highest was found in Echinacea purpurea var.purpurea root with 4137.818ppm and also the lowest in Echinacea purpurea var.double decker root with 528,373 ppm.When the average Fe concentrations of 2013 and 2014 were determined,, the average Fe concentration of 2013 was obtained to be higher with 2358.044ppm.When the average Mn values of the two years were examined, the Mn amount of 2013 with 99,918 ppm was determined higher than Mn amount of 2014 (67.437 ppm).Looking at the average values of roots in 2013 and 2014, the highest Mn value was found in Echinacea purpurea var.purpurea root with 166.595 ppm and the lowest Echinacea purpurea var.double decker root with 49.624 ppm.According to years, the average Pb proportion was found to be higher in 2013 with 2.440 ppm.When the two-year average Pb proportions of roots were studied, Echinacea purpurea var.purpurea root was found the highest with 4,508 ppm.The lowest Pb amount was found in Echinacea paradoxa var.paradoxa root with 1,089 ppm.Looking at the two-year mean values of the roots, the highest Mo mass was found in Echinacea purpurea var.purpurea root as 10.6734 ppm.The lowest Mo amount was obtained from Echinacea purpurea var.baby white swan root with 0.7236 ppm.According to the average of 2013-2014 values, Ni content was found to be higher in 2013 with 5.415 ppm.When the two-year Ni averages of roots were examined, the highest Ni amount was determined to be 9.731 in Echinacea purpurea var.purpurea root.When the average of the two-year values were examined with respect to Zn concentration, it was found to be higher in roots taken at 2013 with 50.068 ppm.When the twoyear average Zn amount of plant roots were considered, the highest was found in Echinacea purpurea var.purpurea root with 96,978 ppm and the lowest in Echinacea purpurea var.double decker root with 23,772 ppm.
The role of each element is different so that plants can nourish and sustain their vital activities.16 of these elements are absolutely necessary for plants, these are; C, H, 0, N, P, K, S, Ca, Mg, Fe, Zn, Mn, Cu, B, CI, and Mo.Some elements are important for some plants, these are; Co, Al, Na, Si, Ni and V.In this study, considering the above information, minerals of Al, Co, Mo, Ca, B, Cd, Cr, Cu, Fe, K, Mn, Na, Ni, P, Pb, S and Zn were determined in Echinacea herbs and roots.Al, Cd, Cr, Co, Cu, Fe, Mn, Pb, Mo, Ni, Zn of these elements are heavy metals and the others (Ca, Na, B, P, S, and K) are important for plants.According to obtained results, it is seen that heavy metals were not found in much amounts in the herbs.Nutrients are taken not only by root systems, but also by above-ground organs, especially by the leaves.Since leaves, stem and flowers are parts of the herb; the mineral proportions we have determined have been found to be higher in Echinacea species with higher leaf density (Echinacea purpurea var.purpurea (L.) Moench, Echinacea pallida var.pallida (Nutt.)Nutt., Echinacea paradoxa var.paradoxa (J.B.S. Norton) Britton, Echinacea purpurea var.baby white swan (L.) Moench and Echinacea purpurea var.double decker (L.) Moench) as seen in Table 1.Heavy metal concentration was determined to be higher in Echinacea roots according to results.Especially Al (aluminum) content were found high.It is thought that the reason of heavy metals to be high in the E. purpurea var.purpurea root is due to the fact that root structure of them are hairy root so the root ends are improved.At the same time, it is thought that the higher levels of heavy metals in the E. purpurea var.purpurea, E. purpurea var.baby white swan and E. purpurea var.double decker roots are due to the hairy root structures and therefore the roots propagate sideways.The heavy metal concentrations were observed lower because they have taproot structure and reach the depths of the soil.

CONCLUSION
Table 1 and Table 2 indicate that there were significant (p<0.01)differences between the herbs and roots of Echinacea species with respect to their mineral compositions.The results of this study demonstrate that the aim of the study to compare of differences between the herbs and roots of Echinacea species.It was observed that the minerals of herbs and roots of Echinacea species were significantly differences.Echinacea spp.contain a lot of the minerals.Furthermore, the mineral composition of Echinacea samples showed a balance as good nutritional quality.However, further detailed studies on the mineral compositons of Echinacea spp are required.In conclusion, the Echinacea spp.contain many valuable minerals that is importance.

Table - 2
. Mean Values and Groupings of Mineral Values of Echinacea Roots (ppm) 5