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Download PDF Researches Upon the Heavy Metals Content Of Current Health Sciences Journal Vol. 36, No. 1, 2010 Original Paper Researches upon the Heavy Metals Content of Rubus caesius L. (Rosaceae) (1) (1) (1) RUXANDRA IONCICĂ , MARIA-VIORICA BUBULICĂ , L. CHIRIGIU , (1) (2) (1,3) G. D. MOGOŞANU , R. POPESCU , H. POPESCU (1)Faculty of Pharmacy, University of Medicine and Pharmacy of Craiova; (2)Faculty of Dental Medicine, University of Medicine and Pharmacy of Craiova; (3)Faculty of Pharmacy, “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca ABSTRACT The analysis of heavy metals in the leaves and stems of Rubus caesius species has been performed. The mean levels of heavy metals in vegetal dried samples were in normal limits. Differences depend on the presence of organic compounds with ligand character and on the environmental conditions. The results indicate that there is not any real danger by utilization of pharmaceutical preparations of Rubii caesii folium. KEY WORDS: Rubus caesius, heavy metals, content. Introduction Rubus caesius L., dewberry, European dew- Material and Methods berry (Rosaceae), is a species native in Europe, including Britain, from Scandinavia south and east Sampling to Spain, Siberia and Western Asia. It is herba- The sampling is a very important phase due to ceous, common in Romania’s flora, along hedge- the specificity of soil, climate, plant developmental rows, amongst shrubs and in roughdry meadow- stages, and collecting time [19]. land, usually on basic soils [1–4]. From the R. caesius species, twenty vegetal In the specialty papers, there are very few data samples were collected at the flowering, in May on the chemical composition and pharmacological 2009, from the surroundings of Bucovăţ Forest, action of the R. caesius species. Dolj County. From the phytochemical point of view, leaves Preparation of plant tissue for analysis and fruits of Rubus species (Rubi idaei folium, Sample preparation is critical in obtaining Rubi fruticosi folium) contain a variety of com- accurate data and reliable interpretation of plant pounds such as tannins, flavonosides (quercetol- analysis results. and camferol glycosides), anthocyanins (in fruits), The vegetal products should be processed pentacyclic triterpene acids (ursolic acid, oleanolic during decontamination, drying, particle-size acid), polyphenolic acids, pectins, organic acids reduction, storage and organic matter destruction. (malic, citric, izocitric, succinic, litospermic, and Plant material must be cleaned and free of oxalic acid), volatile oil (traces), sugars, vitamins extraneous substances, including soil and dust (ascorbic acid), lipids, mineral salts [4–9]. particles that may influence analytical results. Rubus species have some pharmacological The decontamination process must be thorough actions, such as astringent, antidiarrhoeal, mild while still preserving sample integrity. Deconta- laxative, depurative, nutritional, antiseptic, tonic, mination procedures involving washing and haemostatic, diuretic, cicatrizing, anti-inflamma- rinsing with deionized water and 0.2% detergent tory, antioxidant [4, 6–8, 11–18]. solution (non-phosphate), should only be used for For medicinal purposes, leaf infusion and fresh, fully turgid plant samples. After decontami- decoction, syrup and marmalade from the blue- nation, water is removed from plant tissue, at black fruits with sour and astringent taste are used temperatures under 600C, to stop the enzymatic starting from the Rubus species [4, 6–8, 12–18]. reactions and to stabilize the samples. The heavy metals concentrations, depending on Plant tissue samples are reduced to 0.5 to soil properties and other environmental factors, 1.0 mm particle size to ensure homogeneity and represent an indication of quality of medicinal herbs to facilitate organic matter destruction [19]. [19]. The aim of our paper was to analyze the content Gravimetric determination of ash The ash represents the residue obtained of heavy metal cations in the leaves and stems of through the dry ashing of a matter, being made by R. caesius species harvested from the South-West inorganic compounds. of Romania. Ruxandra Ioncică, Pharm., PhD student, University of Medicine and Pharmacy of Craiova 48 Current Health Sciences Journal Vol. 36, No. 1, 2010 Dry-ashing is conducted in a muffle furnace at Results and Discussion 0 temperature 500 to 550 C for four to eight hours. The results of AAS analysis are given as mean For tissues high in carbohydrates and oils, ashing and standard deviation (Table 2, Figure 1). aids may be required to achieve complete decom- position of organic matter. Table 2 – Heavy metals content of R. caesius At the end of the ashing period, the vessel is samples (leaves and stems) Rubii caesii folium Rubii caesii cauli removed from the muffle furnace, cooled, and the Heavy metals ash is dissolved in nitric acid. [mg%] [mg%] 2+ The final solution is diluted as needed to meet Ca 0.9568 ± 0.01 0.2293 ± 0.01 2+ the range requirements of the analytical procedure Zn 4.1302 ± 0.2 3.2157 ± 0.2 Mn2+ 0.0655 ± 0.002 0.1492 ± 0.02 or instrument utilized [19]. 2+/3+ Weigh 0.5 to 1.0 g dried plant material that has Fe 0.8544 ± 0.01 0.1920 ± 0.01 Ni2+ 0.1148 ± 0.01 0.3214 ± 0.01 been ground and homogenized into a high-form, 2+ Pb 1.5462 ± 0.2 4.0084 ± 0.2 30 mL porcelain crucible. Cr3+ 0.1304 ± 0.02 0.0815 ± 0.002 Samples were placed in a cool muffle furnace. sample [g] 1.1279 ± 0.0002 0.6557 ± 0.0002 Temperature control of the furnace was set to dried sample [g] 0.9710 ± 0.0002 0.6094 ± 0.0002 allow gradual increase (two hours) in the ashing ash [g] 0.0848 ± 0.0002 0.0211 ± 0.0002 temperature and maintain for four to eight hours. % dry ashing 8.733 ± 0.5 3.462 ± 0.5 After that, the furnace was turned off to allow residue samples to cool (in one hour). Then, the ash is The maximum limits of heavy metals allowed weighing on analytical balance nearest 0.1 mg. in plants for adequate growing and development 2+ 2+ High temperature oxidation. Heavy metals are the followings: 300 mg% Ca , 10 mg% Zn , 5 mg% Mn2+, 100 mg% Fe2+/3+, 0.80 mg% Ni2+, detection 2+ 3+ This method prepares plant tissue for the 50 mg% Pb and Cr [19]. quantitative determination of the content of Ca2+, In the vegetal products obtained from Rubus Zn2+, Fe2+/3+, Mn2+, Ni2+, Pb2+, Cr3+, by atomic caesius species, the content of the heavy metals can be considered as normal. All samples contain absorption spectrometry (AAS), utilizing high- 2+ 3+ Pb and Cr , markers for soil and air pollution. temperature dry oxidation of the organic matter 2+ and dissolution of the ash with 4% nitric acid. The levels of Pb (normal limits) in vegetal tissues The method detection limit is approximately samples appear because the plants collected were 0.04%. The method is generally reproducible relatively closely to roads (cars circulation). within ±7% [19]. Among the most abundant plant species of mine tailings, dewberry (R. caesius) accumulate Table 1 – Experimental data significant amounts of Pb2+, Cu2+ and Zn2+. Parameters Mo Pb Zn Ni Ca Cr Fe Considering the translocation features, dewberry Wavelength 313.3 283.3 213.9 232.0 422.7 357.9 248.3 accumulates heavy metals primarily in the under- [nm] Lamp current ground organs (roots) [20]. 15 9 13 12 9 10 11 [mA] Generally, for most plants the concentrations Air flow – 590 620 710 630 680 690 of some heavy metals are higher in the roots then [L/hr] Acetylene flow in the aboveground parts. 150 65 65 69 65 65 69 [L/hr] This is an important finding, because only the N2O flow 620 – – – – – – leaves and fruits of R. caesius species are usually [L/hr] used as medicinal products. Flame type Red. Ox. Ox. Ox. Ox. Ox. Ox. High levels of Pb2+, Ni2+ and Mn2+, but in Red. – reducing; Ox. – oxidizing. normal limits, have been determined in stems, The tissue samples (leaves, stems) were because their intake from soil or aerial parts. prepared in the above-mentioned manner. The leaves contain high levels of Ca2+, Zn2+, Heavy metals content was determined using Fe2+/3+ and Cr3+, probably as consequence of the analytical balance, porcelain crucibles, muffle bioinorganic mechanisms during the flowering furnace, volumetric labware, deionized water, period and of the soil acidity. standard calibration solutions, and an AAS–30 AAS analysis confirmed indirectly that the Carl Zeiss Jena (Germany) spectrometer with samples were collected from mature R. caesius Photron & Narva cathode (Table 1). species, during the flowering period, when appear Five standard calibration dilutions (0.001 mg/L complex combinations with divalent (Ca2+, Zn2+, to 2 mg/L) were prepared starting from 5 mg/L Mn2+, Fe2+, Pb2+) or trivalent (Fe3+, Cr3+) metallic reference solutions diluted with 4% nitric acid. cations. 49 Ruxandra Ioncică and colab.: Researches upon the Heavy Metals Content of Rubus caesius L. (Rosaceae) High levels of Mn2+, Ni2+ and Pb2+ have been determined in the stems then in the leaves. The samples have been collected during the flowering period, when appearing complex combinations with divalent (Ca2+, Zn2+, Mn2+, Fe2+, Pb2+) or trivalent (Fe3+, Cr3+) metallic cations. The results indicate that there is not any real danger by utilization of pharmaceutical preparations of Rubii caesii folium. References 1. Ciocârlan V – (2000) Flora ilustrată a României. Pteridophyta et Spermatophyta, Ed. Ceres, Bucureşti, 300. 2. Borza Al – (1968) Dicţionar etnobotanic, Ed. Academiei Române, Bucureşti, 150. 3. Butură V – (1979) Enciclopedie de etnobotanică românească, Ed. Ştiinţifică şi Enciclopedică, Bucureşti, 159. Figure 1 – Maximum limits allowed and heavy 4. Tiţă I – (2003) Botanică farmaceutică, Ed. Didactică metals content of R. caesius samples şi Pedagogică, Bucureşti, 675–676. (leaves and stems). 5. Bruneton J – (1995) Pharmacognosie, Phytochimie, Plantes médicinales, Lavoisier Tec & Doc, Paris, The soil analysis and the avoiding of collecting 333.
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