Application of X-ray fluorescence spectrometry in plant science: Solutions,threats, and opportunities

X-ray fluorescence (XRF) spectrometry is a nondestructive, rapid, simultaneous multi-elemental imaging methodology for plant materials. Its applications are broad and cover most of the elements with varying concentration below the parts per million (ppm). XRF is a well-established atomic spectrometric technique that is also being used as a field portable instrumentation. In recent decades, XRF has been considered a very versatile tool for plant nutrition diagnosis due to its fast and multi-elemental analytical imaging response directly from a solid sample. In this review, we have mainly focused on the recent developments and advancements in XRF spectrometry to analyze plant materials. We have also included the fundamental aspects and instrumentation for XRF spectrometry for its use in plants imaging. We have also covered the use of XRF for vegetal tissues and plant leaves. Mainly, we have briefly focused on some features of sampling procedures and calibration strategies regarding the use of XRF for plant tissues. Microchemical imaging applications by XRF, μ-XRF, μ-SRXRF, and TXRF have been covered for a wide variety of plant tissues such as leaves, roots, stems, and seeds.     

银沙槐不同部位中微量元素含量的测定

利用火焰原子吸收光谱法测定银沙槐的不同部位中Cu、Zn、Ca、Fe、Mg、K、Cr、Mn的含量。对银沙槐植物中的根、茎、叶、花、果实中金属元素的含量进行了分析比较。以此研究了这几种金属元素在银沙槐体内的分布。结果显示,各元素在实验范围内,线性关系良好,加标回收率在92.00%—107.00%之间。相对标准偏差在0.02%—5.80%之间。由实验结果发现,银沙槐的不同部位中Cu、Zn、Ca、Fe、Mg、K、Cr、Mn的含量有差异。     

ICP-AES法测定广西中药白花丹不同部位常量及微量元素

采用电感耦合等离子体发射光谱法(ICP-AES) 分析广西中药白花丹根、茎、叶3个部位的20种常量及微量元素。结果显示,白花丹中除了含人体必需的常量元素K,Ca,Na,Mg外, 还有Zn,Fe,Mn, Cr,Co等必需的微量元素和其他元素Mo, Sb,Bi,Cd, Sr,Pb,Cd,As等。各元素的含量在白花丹的根,茎,叶中分布不同, 在叶和根中的含量比较丰富,与白花丹的抗菌, 抗肿瘤及抗氧化活性的药用有效部位在根部和叶部的结果一致,如Na,K,Ca,Zn,Fe,Mn,Sr,Cu,Co等含量在叶中含量最高,根次之。在许多抗癌中药中,Zn,Mn,Fe的含量通常比较高,Cr,Sr,Cu的含量也至关重要,作为传统抗癌中药的白花丹中就含有丰富的Zn,Mn,Fe和一定量的Sr,Cr,Cu元素,测定结果为探讨中草药中常量、微量元素与生物活性之间的相互协同关系提供了有用的数据,也为更好地开发和利用白花丹药用资源提供理论依据。     

Mapping of the metal intake in plants by large-field X-ray microradiography and preliminary feasibility studies in microtomography

This paper reports on dual energy micro-radiography and tomography techniques applied both to thin plant leaves treated with copper or lead solutions and on Cu-treated small roots and stem sections, performed at the SYRMEP X-ray beamline of ELETTRA synchrotron facility in Trieste (Italy). The features of the source allowed us to apply different imaging techniques with an extremely vast field of view, up to 160 ×6 mm2 and 28×6 mm2 for micro-radiography and tomography experiments, respectively. The feasibility of getting positive indications on metal accumulation in leaves, sections of roots and stems, stem and root whole cylindrical pieces has been checked.     

Direct Analysis of Thallium in Solid Samples of Bovine Liver and Tomato Leaves by Laser Excited Atomic Fluorescence Spectrometry Using a Graphite Furnace

Laser excited atomic fluorescence spectrometry (LEAFS) with a N₂-laser pumped dye laser with a Shimadzu GFA-4A graphite furnace is used to determine thallium in solid samples of bovine liver and tomato leaves. The linear range for thallium under the studied conditions is 0–50 pg. The instrumental LEAFS detection limit for thallium was 100 fg. Solid samples from National Institute of Standards and Technology were analyzed by this method. The content of thallium in bovine liver and tomato leaves was 0.059 and 0.055 μg/g with relative standard deviation of 12.5% and 15.8% respectively; the results obtained are within the reported limit.     

Distribution and utilization of 15N in cowpeas injected into the stem under influence of water deficit

Investigations were carried out on Vigna unguiculata L. Walp. to estimate the distribution and utilization of 15N in different organs after stem injection during vegetative, flowering and pod filling stage. During flowering effects of water deficit were also examined. In well watered plants, within 4 days after injection, 65% of 15N accumulated in leaves. This was drastically reduced to 42% by water deficit. 15N accumulation in stems increased under water deficit. The translocation of 15N from the stem base to roots were not altered by water deficit. During pod filling 62% of recovered 15N in the plants had accumulated in seeds, 24% in leaves and 11% in stems within 4 days, whereas the uptake of nitrogen in pod walls and roots remained low (2%). These results demonstrate that the method of injecting very small quantities (1 mg/plant) of 15N into the stem base allows an exact and detailed quantitative assessment of N translocation/distribution with regard to different organs, different growth stages and different treatments.     

ESEM-EDS: In vivo characterization of the Ni hyperaccumulator Noccaea caerulescens

Environmental scanning electron microscopy (ESEM) permits to analyze samples in their native-hydrated state, allowing a broad spectrum of biological applications. In this study, ESEM equipped with energy dispersive X-ray spectrometer (EDS) was used as a fast method to analyze tissue morphology and to investigate metal distribution in the Ni hyperaccumulator Noccaea caerulescens, an established model to study the adaptation of plants to metalliferous soils. The low vacuum and wet mode operative conditions required the proper choice of experimental parameters both for morphological and compositional characterization of plant tissues. The calibration strategy for semi-quantitative analysis involved the use of Ni fortified agar as standard and signal normalization respect to endogenous carbon, chosen as internal standard. The obtained results are in accordance with present literature, showing a preferential Ni distribution in the epidermal cells respect to near the stomata for leaves and in the cotyledon epidermidis respect to cotyledon parenchyma area for seeds. Thanks to the absence of any time consuming sample treatment steps, ESEM-EDS technique can be proposed as valid strategy for in vivo high-throughput analysis of plant tissues and for a rapid screening and identification of other hyperaccumulator plants in a selected contaminated area.     

美洲商陆对锰毒生理响应的FTIR研究

在一定Mn浓度梯度处理下(0,0.125,0.25,0.5,1 g·kg-1),超积累植物——美洲商陆(Phytolacca americana)不同组织器官的傅里叶变换红外光谱(FTIR)图谱发生了变化。其中,茎组织在3 336和2 916 cm-1处峰高先上升后下降,反映了有机物运输受阻情况的变化,即美洲商陆在低Mn刺激下会产生大量有机物作为渗透调节物质来增强其耐Mn性,高Mn则抑制了有机物的分泌和运输;根和叶组织分别在2 922和1 606 cm-1处表现不同变化趋势,但都反映了一个变化规律即低Mn处理下美洲商陆分泌的有机酸不断螯合Mn,随着Mn毒害的加重,其羧酸螯合力变弱;根组织1 732和1 026 cm-1、茎组织1 028 cm-1、叶组织1 052和967 cm-1处呈现差异性变化,但都与其膜脂过氧化有关;根组织1 375 cm-1处峰高先上升后下降,可能与植物在细胞壁结构上增强抗逆性有一定关系,即低Mn处理下细胞壁可能通过阳离子交换能力(CEC)的提高增强了耐Mn性。以上说明,利用FTIR研究重金属超积累植物化学组分具有应用价值。     

The determination of the parameters of sub- and supercritical extraction of plant raw materials

The extraction of biologically active substances by carbon dioxide from various plant raw materials (amaranth seeds, Sophora japonica flower buds, Stephania rotunda stems, and Stevia rebaudiana leaves) was studied at sub- and supercritical parameters. A laboratory unit for the extraction of plant raw materials by liquefied gases and supercritical fluids at 5–35 MPa pressures and 285–350 K temperatures was developed. The maximum yield of the extracted substances from plants specified was obtained at temperature and pressure exceeding the critical parameters of CO₂ (320–330 K, 28–30 MPa).     

Direct chromium speciation using X‐ray spectrometry and chemometrics

Chromium is a chemical element that occurs in two principal distinct forms: trivalent (Cr(III)) and hexavalent (Cr(VI)). As chromium toxicity depends on its oxidation state, the concentrations of both the species should necessarily be determined, and not only that of total chromium. Using X‐ray spectrometry allied to chemometrics, this work presents a new method for chromium speciation, with no prior separation and/or preconcentration of Cr(III) and Cr(VI) species. Spectra of several solutions containing different concentrations of Cr(III) and Cr(VI) were obtained. The data were treated through principal component analysis and partial least squares regression. The results obtained show that direct speciation using a conventional X‐ray spectrometer is possible, with calculated limits of detection and quantification for Cr(III) and Cr(VI) being lower than 17 and 50 µg g^?1, respectively. Copyright © 2010 John Wiley & Sons, Ltd.     

原子吸收光谱法同时测定中草药内五种元素的研究

微量元素Ｃｕ、Ｚｎ、Ｆｅ、Ｍｎ、Ｃｒ是人体所必需的。本文采用火焰原子吸收光谱法同时测定中草药内这五种微量元素。建立了样品的消化方法及介质、基体对测定影响的抑制措施。通过对野生中草药根、茎、叶和植物标准物的测定，确立了方法的可行性，各元素回收率在９１．３％－１０３．７％，相对标准偏差＜１％。     

Distribution and Utilization of 15N in Cowpeas Injected into the Stem Under Influence of Water Deficit

Abstract

Investigations were carried out on Vigna unguiculata L. Walp. to estimate the distribution and utilization of ¹⁵N in different organs after stem injection during vegetative, flowering and pod filling stage. During flowering effects of water deficit were also examined. In well watered plants, within 4 days after injection, 65% of ¹⁵N accumulated in leaves. This was drastically reduced to 42% by water deficit. ¹⁵N accumulation in stems increased under water deficit. The translocation of ¹⁵N from the stem base to roots were not altered by water deficit. During pod filling 62% of recovered ¹⁵N in the plants had accumulated in seeds, 24% in leaves and 11% in stems within 4 days, whereas the uptake of nitrogen in pod walls and roots remained low (2%). These results demonstrate that the method of injecting very small quantities (1 mg/plant) of ¹⁵N into the stem base allows an exact and detailed quantitative assessment of N translocation/distribution with regard to different organs, different growth stages and different treatments.     

Corroding the chromium cation

Two [Cr,O₂]⁺ isomers have been selectively produced and studied by FT-ICR mass spectrometry. The Cr(O₂)⁺ complex was formed by supersonically expanding a plasma produced by laser vaporization of chromium metal with the helium carrier gas, which was seeded with traces of oxygen, while the chromyl cation is formed in an expansion with N₂O. The complex is stable against thermal collisions, but in a bimolecular reaction with water it is rapidly converted to the chromyl cation, with ligand exchange being only a minor side reaction. Isotopic studies suggest a side-on geometry for Cr(O₂)⁺, in accordance with density functional (B3LYP) calculations. The present work indicates that an investigation of the selected isomers can indeed be carried out, if appropriate chemical methods for the ion generation are applied.     

Characterization of Cr/SiO2 catalysts and ethylene polymerization by XPS

Cr/SiO₂ catalysts with 1 or 3 wt.% Cr loadings and different chromium precursors were characterized by X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). A method to determine chromium species in the sample was developed through the decomposition of the Cr 2p XPS spectrum in Cr⁶⁺ and Cr³⁺ standard spectra. The results of the binding energy from the Cr 2p region and of the distribution of chromium species allowed to evaluate the dynamic photo-reduction of the surface chromium species during XPS analysis. Photo-reduction of surface Cr⁶⁺ to Cr³⁺ species was verified for all samples supported in silica, depending on the precursor and chromium content. Bulk CrO₃ and Cr₂O₃ standards did not reveal variation in the binding energy of Cr 2p_3/2, but a physical mixture of CrO₃ with SiO₂ presented photo-reduction. The behavior of this mixture resembled to the catalysts and suggests the participation of the surface hydroxyls of silica in the photo-reduction process. XPS intensity measurements for assessing dispersion of chromium oxide were used to compare the calcined and reduced catalysts to different chromium precursors. Polyethylene chains were detected by in situ XPS, while oligomerization products were not observed.     

Plumbagin,a Naturally Occurring Naphthoquinone: Its Isolation,Spectrophotometric Determination in Roots,Stems, and Leaves in Plumbago Europaea L.

A new method for isolation and spectrophotometric determination of plumbagin is presented. Plumbagin was isolated by thin layer chromatography (TLC) and column chromatography (CC) techniques, as an orange tinged yellow long crystalline substances. Plumbagin exhibits two absrop-tion maxima at 410 and 510 nm. Stability of the color, pK_a value, and the effect of pH were studied. Beer's law is obeyed over the range 0.9-45 ppm. The method is applied to the determination of plumbagin in roots, stems, and leaves of Plumbago europaea L. plant.     

X‐ray absorption spectroscopy at the Ni?K edge in Stackhousia tryonii Bailey hyperaccumulator

Young plants of Stackhousia tryonii Bailey were exposed to 34 mM Ni kg^?1 in the form of NiSO₄· 6H₂O solution and grown under controlled glasshouse conditions for a period of 20 days. Fresh leaf, stem and root samples were analysed in vivo by micro x‐ray absorption spectroscopy (XAS) at the Ni? K edge. Both x‐ray absorption near edge structure and extended x‐ray absorption fine structure spectra were analysed, and the resulting spectra were compared with spectra obtained from nine biologically important Ni‐containing model compounds. The results revealed that the majority of leaf, stem and root Ni in the hyperaccumulator was chelated by citrate. Our results also suggest that in leaves Ni is complexed by phosphate and histidine, and in stems and roots, phytate and histidine. The XAS results provide an important physiological insight into transport, detoxification and storage of Ni in S. tryonii plants. Copyright © 2008 John Wiley & Sons, Ltd.     

Quantitative Elemental Analysis of Solid Samples by Arf-Excimer Laser-Ablated atomic Emission Spectrometry

Abstract

Quantitative analysis of chromium in standard low alloy steel samples by excimer laser ablated plasma atomic emission spectrometry was obtained. A calibration curve was developed which related the chromium concentration in a solid steel matrix to the intensity ratio of Cr(I) 520.84 nm to Fe(I) 516.75 nm. The chromium concentration determined ranged from 0.062 to 1.31 %. A detection limit of 20 μg/g (approximately 0.002 %) was estimated.     

Thermal dissociation of chromium hexacarbonyl and its decomposition products—Unsaturated carbonyls

The thermal dissociation of chromium hexacarbonyl was studied behind incident shock waves by means of molecular and atomic resonance absorption spectroscopies. The results obtained by these two methods were found to be in close agreement with each other and with the published data on the laser pyrolysis of this compound. A joint interpretation of the experimental results on Cr(CO)₆ dissociation obtained in the present work and the available data on the recombination of CO molecules with unsaturated chromium carbonyls, Cr(CO)₅ and Cr(CO)₄, within the framework of the Rice-Ramsperger-Kassel-Marcus theory made it possible to estimate the bond dissociation energies of all six ligands and to determine, for the first time, the corresponding dissociation rate constants in the low- and high-pressure limits.     

MALDI-based intact spore mass spectrometry of downy and powdery mildews

Fast and easy identification of fungal phytopathogens is of great importance in agriculture. In this context, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool for analyzing microorganisms. This study deals with a methodology for MALDI-TOF MS-based identification of downy and powdery mildews representing obligate biotrophic parasites of crop plants. Experimental approaches for the MS analyses were optimized using Bremia lactucae, cause of lettuce downy mildew, and Oidium neolycopersici, cause of tomato powdery mildew. This involved determining a suitable concentration of spores in the sample, selection of a proper MALDI matrix, looking for the optimal solvent composition, and evaluation of different sample preparation methods. Furthermore, using different MALDI target materials and surfaces (stainless steel vs polymer-based) and applying various conditions for sample exposure to the acidic MALDI matrix system were investigated. The dried droplet method involving solvent evaporation at room temperature was found to be the most suitable for the deposition of spores and MALDI matrix on the target and the subsequent crystallization. The concentration of spore suspension was optimal between 2 and 5?×?10(9) spores per ml. The best peptide/protein profiles (in terms of signal-to-noise ratio and number of peaks) were obtained by combining ferulic and sinapinic acids as a mixed MALDI matrix. A pretreatment of the spore cell wall with hydrolases was successfully introduced prior to MS measurements to obtain more pronounced signals. Finally, a novel procedure was developed for direct mass spectra acquisition from infected plant leaves. Copyright ? 2012 John Wiley & Sons, Ltd.     

Effect of hydrazine and hydroxylaminophosphate on chrome plating from trivalent electrolytes

The influence of hydrazine and hydroxylaminophosphate on solution chemistry and quality of chromium coatings deposited from Cr(III) formate-urea baths has been studied by FT-IR, XPS and AFM. The results have shown that hydrazine and hydroxylaminophosphate differently affect Cr plating. The morphology of Cr deposits obtained in the formate-urea electrolytes is a typical nodular structure with a less nodular and smoother fine-grained structure of Cr obtained in the presence of hydroxylaminophosphate. Hydrazine, in contrast, worsened the quality and lowered the hardness of the deposits. The current efficiency of chromium increases in the presence of both hydrazine and hydroxylaminophosphate. This may be related with the ability of these ligands to reduce the CrH content in the growing Cr deposit. The data obtained suggest that hydroxylaminophosphate improves the stability of the active [Cr(carbamide)_n(H₂O)_6−n]³ complexes providing good quality of the deposits and prolonged working lifetime in the formate-urea bath.