Assessment of Heavy Metals in Pharmacological Important Medicinal Plants Consumed in the Bannu District,Pakistan

This study assesses the contents of heavy metals in the four most commonly used indigenous medicinal plants in the Bannu District, Pakistan. The rapid appraisal approach was used along with semi-structured interviews with elderly people and herbalists for selection of indigenous plants. Heavy metals were determined via flame atomic absorption spectrometer by acid digestion of samples. Four medicinal plants were selected after interviews of 53 local residents and herbalists. Plant and soil samples were obtained for evaluations from a wastewater zone (WWZ) and a clean water zone (CWZ). Indigenous plants from the WWZ showed considerably higher metal contents compared to the CWZ. The trend of metals for indigenous medicinal plants grown in clean water was Mn > K > Na > Zn > Co > Fe > Cu while for wastewater the trend appeared as Co > K > Na > Zn > Fe > Mn > Cu. An alternative significant extent of research is needed to qualify the assessment of the human health insinuations of consumption of indigenous remedies. The heavy metal content of administering herbal medicine should be screened, as formulation and processing of medicine may affect heavy metal contents of the remedies.     

Direct and rapid determination of ultratrace heavy metals in solid plant materials by ET‐AAS ultrasonic‐assisted slurry sampling

Introduction – Plants can be used as bioindicators in the study of contamination processes by heavy metals. Most of the analytical methodologies used for determination of metals in plants are based on atomic techniques with previous wet digestion of the solid samples. Methodologies that allow direct metal measurements in solid samples are very attractive alternatives. Objective – To develop a new procedure for direct analysis of copper, nickel, cadmium and lead at very low concentration levels in leaves based on electrothermal atomic absorption spectroscopy (ET‐AAS) with introduction of samples as a slurry. Methodology – In order to obtain accurate and precise results even at very low concentrations, the different parameters that influence the sample slurry preparation such as acid percentage, presence of stabilising agents and ultrasonic probe operation were studied. Instrumental parameters such as chemical modifier and temperature and times for drying, pyrolysis and atomisation steps that influence ET‐AAS measurement were optimised. Results – Optimal slurry conditions for copper and nickel determination were 0.5% Tween 85 with 5% nitric acid. For lead and cadmium analysis the best results were obtained in 5% nitric acid without stabilising agents. The achieved detection limits were 0.023 mg/kg for copper, 0.018 mg/kg for nickel, 0.0002 mg/kg for cadmium and 0.009 mg/kg for lead. For validation purposes, the method was applied to metal analysis in a pine needles reference material. Conclusion – According to our knowledge, the detection limits obtained are the best reported in the literature. The methodology was successfully used in metal determinations in actual leaf samples. Copyright © 2010 John Wiley & Sons, Ltd.     

Analysis of trace elements in animal tissues

Graphite furnace atomic absorption spectrophotometry is a method used for the measurement of low concentrations of manganese (ppb range). Despite the widespread use of this technique, there is considerable inconsistency concerning sample preparation and choice of instrumental parameters. In this paper, we determined manganese concentrations of National Bureau of Standards (NBS) bovine liver by both graphite furnace (Instrumentation Laboratory IL 555B) and flame atomic absorption following wet digestion of the sample with nitric acid. The following instrumental parameters for the graphite furnace were found optimal for the measurement of manganese in digested NBS bovine liver: inert gas flow=14 SCFH, drying temperature 100°C/15 s (step 1), 125°C/15 s (step 2), pyrolysis temperature 500°C/15 s (step 3), and 1000°C/15 s (step 4); atomization temperature 2250°C/10 s (step 5). For optimal results, the nitric acid concentration of the sample should be between 2 and 4M. There were no significant differences found for manganese concentrations determined by either peak height or peak area measurement. Additionally, no significant differences were found in manganese concentrations determined by flame or furnace methods. Assuming proper sample preparation and choice of instrumental parameters, values obtained for manganese concentration by graphite furnace and flame atomic absorption spectrophotometry are similar. Therefore, data obtained by these two methods can be compared directly.     

Chromium in plants

Chromium is an essential trace element and is associated with some biological pathways, especially with glucose tolerance. For these reasons, we decided to determine the concentration of chromium in two sets of Brazilian medicinal plants. The first group consisted of plants that are considered as antidiabetic, whereas the second included plants that do not have this therapeutic property. The concentration of chromium was determined by flameless atomic absorption. All the plants analyzed contain chromium in the normal range for this element, but the hypoglycemic plants contain more chromium than the others (1–4 μg/g compared to 0.5–1.5 μg/g).     

Concentration‐ and flux‐based dose–responses of isoprene emission from poplar leaves and plants exposed to an ozone concentration gradient

Concentration‐ and flux‐based O₃ dose–responses of isoprene emission from single leaves and whole plants were developed. Two poplar clones differing in O₃ sensitivity were exposed to five O₃ levels in open‐top chambers for 97 d: charcoal‐filtered ambient air (CF), non‐filtered ambient air (NF) and NF plus 20 ppb (NF + 20), 40 ppb (NF + 40) and 60 ppb (NF + 60). At both leaf and plant level, isoprene emission was significantly decreased by NF + 40 and NF + 60 for both clones. Although intra‐specific variability was found when the emissions were up‐scaled to the whole plant, both leaf‐ and plant‐level emissions decreased linearly with increasing concentration‐based (AOT40, cumulative exposure to hourly O₃ concentrations >40 ppb) and flux‐based indices (POD_Y, cumulative stomatal uptake of O₃ > Y nmol O₃ m^?2 PLA s^?1). AOT40‐ and POD₇‐based dose–responses performed equally well. The two clones responded differently to AOT40 and similarly to POD_Y (with a slightly higher R² for POD₇) when the emission was expressed as change relative to clean air. We thus recommend POD₇ as a large‐scale risk assessment metric to estimate isoprene emission responses to O₃ in poplar.     

郑汴路路旁土壤-小麦系统重金属积累及其健康风险评价

按离路基不同距离采集土壤、麦苗和籽粒样品, 在测定样品重金属 (Pb、Cd、Zn、Cr和Cu) 的基础上, 开展了路旁土壤-小麦系统重金属分布、积累和污染状况分析, 并对膳食小麦 (Triticumaestivum) 引起的健康风险进行了评价。结果表明:1) 土壤-小麦系统重金属含量随着离开路基距离的增加呈先增加后减少的趋势, 土壤重金属含量>麦苗重金属含量>籽粒重金属含量。2) 麦苗和小麦籽粒对土壤重金属富集能力的大小顺序均为Cu>Cd>Zn>Pb>Cr, 麦苗对重金属的富集能力大于小麦籽粒。3) 膳食小麦所致的Cd个人健康风险较大。     

Health risk assessment by toxic metals in little egrets (Egretta garzetta) and food chain contaminations

This study analysed heavy metals from little egret (Egretta garzetta). Egret’s Eggs, egg shells, food (fish and insects), blood, meat samples (thigh, liver, and chest), water, soil and sediments samples were collected from the two selected sites of the study area. Samples were analysed on flame atomic absorption spectrometer after acid digestion. Detected metals were found almost inline of concentrations when compared with the both sites. Among detected metals Mn was found higher in concentration (µg/g) i.e. 18.509 followed by Zn i.e. 9.383, Ni, Cu, Pb and Cd. Sediment exhibited higher levels (µg/g) of metals (25.061) followed by the meat (19.044) egrets food (18.825), excreta (16.26), blood serums (4.577), eggs (3.626) and water samples (2.432).The level of metals in sediments of the study are showed environmental concerns. Health risks were also investigated that were compared to guidelines of WHO and FAO threshold limits. It was found a marginal health risk to life through detected metals. This study revealed that little egret are good bio-indicator for the screening and investigation of contaminates presence in the environment.     

An integrating atomic absorption spectrophotometer for ultramicroanalysis of metals

An integrating atomic absorption spectrophotometer for ultramicroanalysis of metals is described. The apparatus is intended mainly for determination of calcium and magnesium but can also be used for sodium, potassium, lithium, and copper, among other metals. Analyses of sample volumes as small as about 1 nl, containing calcium and magnesium in amounts as low as 10^?11 and 10^?12 moles, respectively, have been made. In analyses of samples containing 4·10^?12 moles of magnesium a coefficient of variation of ±5% was obtained.The principles of the apparatus are the same as for a conventional atomic absorption spectrophotometer apart from the fact that an electronic integrator with a rapid response has been used as the recording system. For atomization of the sample a hydrogen-air flame has been used in combination with a flame adapter. The sample is introduced into the flame by means of a small loop on a platinum-iridium filament.     

Utilizing a nano‐sorbent for the selective solid‐phase extraction of vanillic acid prior to its determination by photoluminescence spectroscopy

Vanillic acid (VA) is a phenolic acid, and acts as a natural antioxidant in fruits, vegetables and plants. The extraction and determination of trace levels of VA in plants is important, because stimulation of protein synthesis and activation of antioxidant enzymes occur in the presence of phenolic acids at trace levels. In this research, a photoluminescence spectroscopic method was developed for the quantification of VA in plant samples after separation and pre‐concentration. Selective extraction of VA from aqueous solution was performed using a solid‐phase extraction column packed with nickel–aluminum layered double hydroxide as a nano‐sorbent. After elution of extracted analyte from the column using 3 mL of a 3 mol/L NaOH solution, its concentration was determined spectrofluorometrically at λ_em = 357 nm with excitation at λ_ex = 280 nm. The spectrofluorometry method gave a linear response for VA within the range 20.0–900.0 µg/L, with a correlation coefficient of 0.9982. The limit of detection and sorption capacity were 7.6 µg/L and 66.2 mg/g, respectively. The method was validated by comparing the obtained results with gas chromatographic data. This method was used to determine VA in Chenopodium album and Prangos asperula plants. Copyright © 2014 John Wiley & Sons, Ltd.     

Rapid method for simultaneous determination of 20 components in Isodon nervosa by high‐performance liquid chromatography–electrospray ionisation tandem mass spectrometry

Introduction – Isodon nervosa is a commonly used traditional Chinese medicine including diterpenoids, phenolic acids, triterpenoids and volatile oil. Qualitative and quantitative analysis of multi‐components is important for its quality control. Objective – To establish a liquid chromatography–electrospray ionisation–mass spectrometry method for simultaneous analysis of 20 bioactive constituents of Isodon nervosa in different places of China and different parts of this herb. Methodology – The optimal chromatographic conditions were achieved on a C₁₈ column (250 × 4.6 mm, 5 µm) with with linear gradient elution with 0.1% aqueous formic acid : methanol containing 0.1% formic acid at a flow‐rate of 0.7 mL/min in 15 min. The identification and quantification of those analytes were achieved on a hybrid quadrupole linear ion trap mass spectrometer. Multiple‐reaction monitoring scanning was employed for quantification with switching electrospray ion source polarity between positive and negative modes in a single run. Full validation of the method was carried out (linearity, precision, accuracy, limit of detection and limit of quantification). Results – The results indicated that the method was simple, rapid, specific and reliable. The proposed method was successfully applied for the qualitative and quantitative analysis of 20 chemical compositions in Isodon nervosa samples. Conclusion – Twenty chemical compositions in 21 batches of wild and cultivated Isodon nervosa samples from different sources had great variation in the contents. Copyright © 2010 John Wiley & Sons, Ltd.     

Use of biomonitors for the identification of heavy metals emission sources

Heavy metals accumulation by Tillandsia capillaris was measured to identify their main emission sources in the province of Córdoba, Argentina. Samples of T. capillaris collected over three years at different sites were analysed by flame atomic absorption spectrometry to determine the amounts of Cu, Fe, Ni, Mn, Pb and Zn. The sampling sites were categorized according to land use, anthropic activities and/or distance from the potential heavy metal emission sources. We found that the concentration of heavy metals in the study area is mainly driven by industrial activity while traffic contributed only to the levels of Zn. In addition, we observed a strong relationship between a diffuse emission source and the content of Pb accumulated on the biomonitors which could be attributed to dove hunting activities. Future studies are needed to confirm this hypothesis.     

Analytical methods for heavy metals in herbal medicines

Introduction – It is estimated that about 70–80% of the world's population relies on non‐conventional medicine, mainly of herbal origin. However, owing to the nature and sources of herbal medicines, they are sometimes contaminated with toxic heavy metals such as lead, arsenic, mercury and cadmium, which impose serious health risks to consumers. It is critical to analyse source materials for heavy metals in order to ensure that their concentrations meet the related standards or regulations limiting their concentrations in herbal medicines. In this review, different analytical methods for analysis of heavy metals in herbal medicines are discussed. Objective – To provide a comprehensive review of the current state of the art in analytical methods used to detect heavy metals in herbal medicines. Methodology – We systematically searched and reviewed the research articles regarding analytical methods for heavy metals in herbal medicine from various databases, such as Medline/PubMed, ScienceDirect, SciFinder, Google Scholar, EBSCO, Gale InfoTrac, Ingenta, Ovid, ProQuest and ISI Web of Knowledge. Results – In this review, we discuss in detail several commonly used and sensitive analytical techniques, including atomic absorption spectrometry, inductively coupled plasma optical emission spectrometry or mass spectrometry, X‐ray fluorescence spectrometry, high‐performance liquid chromatography, differential pulse polarography, neutron activation analysis and anodic stripping voltammetry. We also provide some application examples of these analytical techniques for heavy metals in herbal medicines. Copyright © 2011 John Wiley & Sons, Ltd.     

A RP‐HPLC‐DAD‐APCI/MSD method for the characterisation of medicinal Ericaceae used by the Eeyou Istchee Cree First Nations

Introduction – Ericaceae medicinal plants are traditionally used by the Eeyou Istchee Cree and other northern peoples of North America to treat type 2 diabetic symptoms. Because of the importance of phenolics as potential cures for degenerative diseases including type 2 diabetes, an analytical method was developed to detect them in the leaf extracts of 14 Ericaceae plants. Objective – To develop an optimised method which is applicable to a relatively large number of Ericaceae plants using their leaf extracts. For this purpose phenolics with a wide range of polarity, including a glucosylated benzoquinone, two phenolic acids, three flavanols, a flavanone, a flavone and five flavonols, were included in this study. Methodology – Characterisation of phytochemicals in extracts was undertaken by automated matching to the UV spectra to those of an in house library of plant secondary metabolites and the authentication of their identity was achieved by reversed phase‐high‐performance chromatography–diode array detection–atmospheric pressure chemical ionisation/mass selective detection. Results – Twenty‐six phenolics were characterised within 26 min of chromatographic separation in 80% ethanol extracts of 14 Ericaceae plants. The calibration curves were linear within 0.5–880 µg/g dry mass of the plant with regression values better than 0.995. The limits of detection ranged from 0.3 for µg/mL for (+)‐catechin to 2.6 µg/mL for chlorogenic acid. This is a first study dealing with relatively large number of Ericaceae extracts and is applicable to other plants of same family. Copyright © 2010 John Wiley & Sons, Ltd.     

Accumulation pattern of endogenous cytokinins and phenolics in different organs of 1‐year‐old cytokinin pre‐incubated plants: implications for conservation

A better understanding of phytohormone physiology can provide an essential basis to coherently achieve a conservation drive/strategy for valuable plant species. We evaluated the distribution pattern of cytokinins (CKs) and phenolic compounds in different organs of 1‐year‐old greenhouse‐grown Tulbaghia simmleri pre‐treated (during micropropagation) with three aromatic CKs (benzyladenine = BA, meta‐topolin = mT, meta‐topolin riboside = mTR). The test species is highly valuable due to its medicinal and ornamental uses. Based on UHPLC‐MS/MS quantification, mT and mTR pre‐treated plants had the highest total CK, mostly resulting from the isoprenoid CK‐type, which occurred at highest concentrations in the roots. Although occurring in much lower concentrations when compared to isoprenoid CKs, aromatic CKs were several‐fold more abundant in the root of mT pre‐treated plants than with other treatments. Possibly related to the enhanced aromatic CKs, free bases and ribonucleotides, plants pre‐treated with mT generally displayed better morphology than the other treatments. A total of 12 bioactive phenolic compounds, including four hydroxybenzoic acids, five hydroxycinnamic acids and three flavonoids at varying concentrations, were quantified in T. simmleri. The occurrence, distribution and levels of these phenolic compounds were strongly influenced by the CK pre‐treatments, thereby confirming the importance of CKs in phenolic biosynthesis pathways.     

Microwave-assisted acid extraction methodology for trace elements determination in mastic gum of Pistacia lentiscus using inductively coupled plasma atomic emission spectrometry

Introduction – To ensure food safety, accurate knowledge of the levels of several trace elements is necessary. This is also true for natural products of plants and resins used for human consumption or therapeutic treatment, like the mastic gum of Pistacia lentiscus. The rapid analysis of gum and resin matrices is a challenge because there are problems with the decomposition of such complicated matrices. Objective – To develop an efficient multielemental analytical method for the determination of trace elements and to compare different procedures for analyte extraction when microwave‐assisted digestion is applied. Methodology – The inductively coupled plasma atomic emission spectrometric (ICP‐AES) technique was applied and the optimum ICP conditions like radiofrequency power, argon flow rate and nebuliser sample uptake flowrate were found. The microwave‐assisted procedure was compared with that with conventional heating. Since mastic and resinous materials are difficult for dissolution and extraction of trace element, influential acid mixtures containing hydrofluoric acid proved to be capable of quantitative extraction of the analytes. Results – The digestion of mastic resin or similar matrices is significantly facilitated by using microwave radiation instead of conventional heating since the obtained recovery for several analytes is much higher. It was proved that the acid mixture of HCl–HNO₃–HF was the most efficient for complete sample digestion and recovery of the analytes. Conclusions The performance characteristics of the developed method were evaluated against certified reference material and the method was proved reliable and applicable to the analysis of mastic gum and possibly to similar resinous matrices. Copyright © 2010 John Wiley & Sons, Ltd.     

Pre‐steady state kinetics of ATP hydrolysis by Na,K‐ATPase

Fast reaction kinetics of ATP hydrolysis by Na,K‐ATPase has been investigated by following absorption pattern of pH sensitive dye in stopped flow spectrophotometer. Distinct pre‐steady state phase signal could be recorded with an initial decrease in acidity followed by increase in acidity. Average half time for H⁺ absorption and peak alkalinity was, respectively, 30 ms and 60 ms. Under optimal Na⁺ (120 mM) and K⁺ (30 mM) concentrations, magnitude of both H⁺ absorption and H⁺ release are found to be about 1.0 H⁺/ATPase molecule. H⁺ absorption and release decreased with decrease in Na⁺ concentration, H⁺ release was more affected. Both H⁺ absorption and H⁺ release are found to be independent of K⁺ concentration in the pre‐steady state phase. No H⁺ absorption or release was observed following mixing of either ADP, Na⁺ or K⁺ alone with ATPase. Effect of delayed mixing of Na⁺ or K⁺ on two phases of pre‐steady state cycle indicates that ATP hydrolytic cycle starts without K⁺ ions if optimal Na⁺ is present. ATP hydrolytic cycle does not start in the absence of Na⁺ ions. Results obtained have been interpreted in terms of an extended kinetic scheme for Na,K‐ATPase. Copyright © 2009 John Wiley & Sons, Ltd.     

公路绿化植物油松(Pinus tabulaeformis)和小叶杨(Populus simonii)对重金属元素的吸收与积累

对内蒙古西部公路绿化植物油松（Pinus tabulaeformis）、小叶杨（Populus simonii）及其根际土壤中重金属元素（Cd、Hg、Pb、Cu、Zn、Ni、Cr）和类金属元素（As和Se）含量以及根际土壤重金属（Cu、Zn、Pb、Ni和Cr）形态、土壤pH值进行了测定。对比分析了公路沿线不同绿化植物及其不同器官对重金属元素的吸收与积累特征。结果表明：绿化植物根际土壤对重金属元素的吸附及污染程度以Cd为最高。随原子序数的递增,小叶杨和油松两种植物的根部和茎叶两种营养器官中重金属的含量均表现出“N”字形变动趋势。而且重金属元素在不同植物不同器官中的含量具有Zn〉Cu〉Ni,Cr,As,Pb〉Cd〉Hg的基本规律。小叶杨茎叶对重金属元素Cr、Ni和Pb的富集能力较根部为强,油松茎叶对重金属元素Cr、Ni、Cu和Pb的富集能力较根部为强。绿化植物根际土壤重金属元素有效态占总量百分比的大小序列为Zn〉Pb〉Ni、Cr〉Cu,与重金属元素在不同植物不同器官中的含量大小序列Zn〉Cu〉Ni、Cr、As、Pb〉Cd〉Hg并非趋于一致。公路绿化植物对根际土壤中重金属元素的吸收和积累与重金属元素有效态所占的比例有关。     

Assessment of some elements in human permanent healthy teeth,their dependence on number of metallic amalgam fillings,and interelements relationships

In this study, 60 human permanent healthy teeth (without filling) were collected postmortem and analyzed using flame atomic absorption spectrometry (FAAS), cold vapor atomic absorption spectrometry (CVAAS), hydride generation atomic absorption spectrometry (HGAAS), and electrothermal atomic absorption spectrometry (ETAAS) for the determination of Ca, Hg, Se, Cu, and Ag. The concentration of these elements was assessed in carious and noncarious teeth, different tooth groups, with age and with number of amalgam fillings. A negative correlation was found between Ca and the number of amalgam fillings, and significant negative correlations were found between Ca and three other metals (Hg, Ag, and Cu) that indicate the possibility of substitutions of Ca by three other metals. Significant positive correlations were found among the number of amalgam fillings and Hg, Ag, Cu and Se showed metal concentration in permanent healthy teeth were affected by the presence of the number of amalgam filling. In addition, significant positive correlations between Hg and Ag, Hg and Cu, and Ag and Cu proved the suspicion that the Hg content in permanent healthy teeth was mainly found because of the influence of amalgam filling, not from other sources. Moreover, the significant positive correlation between Hg−Se and Ag−Se showed the formation of mercuric selenide and silver selenide complexes as part of a natural mechanism of detoxification. Consequently, the permanent healthy teeth would be considered as a bioindicator for the accumulation of long-term exposure of Hg and Ag.     

When condition trumps location: seed consumption by fruit‐eating birds removes pathogens and predator attractants

Seed ingestion by frugivorous vertebrates commonly benefits plants by moving seeds to locations with fewer predators and pathogens than under the parent. For plants with high local population densities, however, movement from the parent plant is unlikely to result in ‘escape’ from predators and pathogens. Changes to seed condition caused by gut passage may also provide benefits, yet are rarely evaluated as an alternative. Here, we use a common bird‐dispersed chilli pepper (Capsicum chacoense) to conduct the first experimental comparison of escape‐related benefits to condition‐related benefits of animal‐mediated seed dispersal. Within chilli populations, seeds dispersed far from parent plants gained no advantage from escape alone, but seed consumption by birds increased seed survival by 370% – regardless of dispersal distance – due to removal during gut passage of fungal pathogens and chemical attractants to granivores. These results call into question the pre‐eminence of escape as the primary advantage of dispersal within populations and document two overlooked mechanisms by which frugivores can benefit fruiting plants.     

Contents of macro-, microelements and long-lived radionuclides in the medicinal plants belonging to the wetland community of Siberian region,Russia

Studied contents of macro-, microelements and long-lived radionuclides in the following medicinal plants belonging to the wetland community of Siberian region, Russia: Sphagnum fuscum (Schimp.) Klinggr., Sphagnum balticum (Russ.) Russ. ex С. Jens., Polytrichum strictum Brid., Pinus sylvestris L., Pinus sibirica Du Tour, Betula pubescens Ehrh., Andromeda polifolia L., Ledum palustre L., Oxycoccus palustris Pers., Rubus chamaemorus L., Vaccinium uliginosum L., Comarum palustre L., Chamaedaphne calyculata (L.) Moench. Samples were collected in the sphagnum wetlands of Tomsk region and Khanty-Mansi Autonomous Okrug. For the purpose of research, we used roots, shoots and bark of the plants in the Pinophyta and Magnoliophyta species and turf of the Bryophyta species. The content of macro-, microelements in the plants was determined by an atomic emission spectrometer with inductively coupled plasma “iCAP 6300 Duo” of the “Thermo Scientific” company. The specific activities of the long-lived radionuclides Th-232, K-40, Ra-226, Cs-137 were measured by a gamma-ray spectrometric complex with a high purity germanium semiconductor detector by “ORTEC” (AMETEK) and a digital analyser “ORTEC DSPEC LF”. The obtained results of the element content are useful when selecting a medicinal plant species (or part thereof) which may be used in preparation of new medicinal drugs.