Authentication of herbal drug Tukhm‐e‐balango (Lallemantia royleana Benth.) using microscopic,pharmacognostic, and phytochemical characterization

The study is aimed to provide a comprehensive account on authentication of herbal drug named as Tukhm‐e‐balango (Lallemantia royleana Benth.) from the seeds of Ocimum basilicum by using microscopic, pharmacognostic, and phytochemical characterization. The crude medicinal plants and their parts are often adulterated or substituted in market due to improper identification by the consumers while among herbal plant sellers, taxonomic confusion is caused due to morphological similarities of the plant parts and lack of a standard identification system.In microscopy, both herbarium and fresh specimens were studied using qualitative and quantitative morphological characteristics of leaves, seeds, and pollen. For pharmacognosy, solubility, fluorescence, and physicochemical characterizers were analyzed whereas a total phenolic and flavonoids contents was determined in addition to DPPH radical scavenging activity. In current study, microscopic, pharmacognostic, and phytochemical characterization clearly differentiated L. royleana from O. basilicum. The major problem in herbal drug industry is caused due to confusion and controversy of certain synonyms used for more than one or two drugs. Sometimes, under the same common or local name, entirely different taxa are being sold in herbal markets. It is concluded that correct and proper identification of medicinal plants is very crucial to ensure the safety and efficacy of herbal medicines, as many medicinal plants are intentionally or unintentionally adulterated with similar species or varieties. In herbal market, the seeds of L. royleana are adulterated with seeds of O. basilicum due to their similar morphology.     

Comparative SEM and LM foliar epidermal and palyno‐morphological studies of Amaranthaceae and its taxonomic implications

Palynological features as well as comparative foliar epidermal using light and scanning electron microscope (SEM) of 17 species (10genera) of Amaranthaceae have been studied for its taxonomic significance. Different foliar and palynological micro‐morphological characters were examined to explain their value in resolving the difficulty in identification. All species were amphistomatic but stomata on abaxial surface were more abundant. Taxonomically significant epidermal character including stomata type, trichomes (unicellular, multicellular, and capitate) and epidermal cells shapes (polygonal and irregular) were also observed. Pollens of this family are Polypantoporate, pores large, spheroidal, mesoporous region is sparsely to scabrate, densely psilate, and spinulose. All these characters can be active at species level for identification purpose. This study indicates that at different taxonomic levels, LM and SEM pollen and epidermal morphology is explanatory and significant to identify species and genera.     

Palyno‐morphological characteristics of gymnosperm flora of pakistan and its taxonomic implications with LM and SEM methods

The present study is intended to assess gymnosperms pollen flora of Pakistan using Light Microscope (LM) and Scanning Electron Microscopy (SEM) for its taxonomic significance in identification of gymnosperms. Pollens of 35 gymnosperm species (12 genera and five families) were collected from its various distributional sites of gymnosperms in Pakistan. LM and SEM were used to investigate different palyno‐morphological characteristics. Five pollen types (i.e., Inaperturate, Monolete, Monoporate, Vesiculate‐bisaccate and Polyplicate) were observed. Six In equatorial view seven types of pollens were observed, in which ten species were sub‐angular, nine species were Traingular, six species were Perprolate, three species were Rhomboidal, three species were semi‐angular, two species were rectangular and two species were prolate. While five types of pollen were observed in polar view, in which ten species were Spheroidal, nine species were Angular, eight were Interlobate, six species were Circular, two species were Elliptic. Eighteen species has rugulate and 17 species has faveolate ornamentation. Eighteen species has verrucate and 17 have gemmate type sculpturing. The data was analysed through cluster analysis. The study showed that these palyno‐morphological features have significance value in classification and identification of gymnosperms. Based on these different palyno‐morphological features, a taxonomic key was proposed for the accurate and fast identifications of gymnosperms from Pakistan.     

Morpho‐palynological study of Cyperaceae from wetlands of Azad Jammu and Kashmir using SEM and LM

In this study 12 species of Cyperaceae have been studied for quantitative and qualitative observation of pollen grains through Light and scanning electron microscopy. Pollens of 12 species of Cyperaceae from different wetlands of Azad Jammu and Kashmir were collected. Morphological characters of pollen grains were then investigated under the Light and Scanning electron microscope. Two pollen types have been observed apolar and heteropolar. Shape of pollens was prolate (4 spp), sub‐spheroidal (7 spp), and oblate (1 spp). Variation observed in exine sculpturing granular (4 spp), reticulate (1 spp), areolate‐punctate (3 spp), and psilate (2 spp). Polar to equatorial ratio and fertility percentage of the pollens were also studied. Based on these micromorphlogical characters of pollens taxonomic keys have been made for the accurate identification of the members of Cyperaceae. The characteristics studied in present research work are very much valuable taxonomically and phytochemically for the identification of species of family Cyperaceae. Light microscope (LM) and Scanning electron microscope (SEM) were used for pollen observation, which play vital role in the taxonomical identification of species and provide sufficient information for taxonomist.     

Scanning Electron Microscopy (SEM) and Light Microscopy (LM)‐based Palyno‐morphological views of Solanaceae in Western Himalaya

In this study, plants belonging to family Solanaceae growing in Western Himalaya region have been observed palynologically under Light Microscope and Scanning electron microscope. Present investigation comprises of 10 genera and 23 species, namely, Atropa acuminata, Capsicum decoraticus, Capsicum frutescens, Cestrum aurantiacum, Cestrum diurnum, Cestrum nocturnum, Datura alba, Datura innoxia, Datura stramonium, Hyoscymus niger, Lycopersicon esculentum, Nicotiana rustica, Nicotiana tabacum, Petunia alba, Petunia hybrida, Solanum erianthum, Solanum melongena, Solanum miniatum, Solanum pseudocapsicum, Solanum surratense, Solanum tuberosum, Withania coagulans, Withania somnifera. Solanaceae is a eurypalynous family. Grains are usually Tricolporate and Tetracolporate, radially symmetrical, isopolar, prolate‐spheroidal to oblate‐spheroidal to oblate‐spheroidal to subprolate to per prolate or suboblate to oblate, size range: 8.55–72 μm, amb circular, semi‐angular or subangular, aperture drop‐type, labrum common‐type, exine usually 2 μm thick, nexine 1–1.5 μm thick. Tectum usually psilate, sexine reticulate, granulate or striato‐reticulate, with obscure pattern, sexine 1–2 μm thick, nexine 1–1.5 μm thick, and intine 0.5–1 μm thick. Most striking variation has been found in the shape class, aperture‐type, and tectal surface. Based on these characters, taxonomic keys have been made for correct identification of members in Solanaceae. However, the grains of this family are usually tricolporate and have direct relationship with certain members of the family Scrophulariaceae. Palyno‐morphological characters of family Solanaceae have been studied for the first time in Western Himalayan region of Pakistan. These palyno‐morphological characters are significant for identification of the members of family Solanaceae.     

Identification of powdered Chinese herbal medicines by fluorescence microscopy,Part 1: Fluorescent characteristics of mechanical tissues,conducting tissues,and ergastic substances

The light microscope has been successfully used in identification of Chinese herbal medicines (CHMs) for more than a century. However, positive identification is not always possible. Given the popularity of fluorescence microscopy in bioanalysis, researchers dedicated to finding new ways to identify CHMs more effectively are now turning to fluorescence microscopy for authentication purposes. Some studies on distinguishing confused species from the same genus and on exploring distributions of chemicals in tissues of CHMs by fluorescence microscopy have been reported; however, no systematic investigations on fluorescent characteristics of powdered CHMs have been reported. Here, 46 samples of 16 CHMs were investigated. Specifically, the mechanical tissues including stone cells and fibers, the conducting tissues including three types of vessels, and ergastic substances including crystals of calcium oxalate and secretions, in various powdered CHMs were investigated by both light microscope and fluorescence microscope. The results showed many microscopic features emit fluorescence that makes them easily observed, even against complex backgrounds. Under the fluorescence microscope, different microscopic features from the same powdered CHM or some same features from different powdered CHMs emitted the different fluorescence, making this information very helpful for the authentication of CHMs in powder form. Moreover, secretions with unique chemical profiles from different powdered CHMs showed different fluorescent characteristics. Hence, fluorescence microscopy could be a useful additional method for the authentication of powdered CHMs if the fluorescent characteristics of specific CHMs are known. Microsc. Res. Tech. 2010. © 2010 Wiley‐Liss, Inc.     

Morphological characterization of Hypnaceae (Bryopsida,Hypnales): Investigating four genera from Western Himalayas by using LM and SEM techniques

The Hypnaceae is one of the largest and diversified family among the pleurocarpous mosses which consists of nearly 60 genera and 1,000 species around the world. In Pakistan, it is represented by 15 species and 8 genera. The current research article provides the detail morphological investigation of four different species (genera) of the family Hypnaceae that is, Gollania clarescens, Hypnum revolutum, Homomallium andoi, and Taxiphyllum taxirameum in the Western Himalayas, Pakistan. The research article reports the first SEM study of the leaf surfaces of the studied taxa, along with comprehensive morphological characters of the four species. The main objective of the research project is to present the comparative light and scanning electron microscopic study to discuss the morphology in detail because previously the family is just reported in different bryophyte checklists of the Western Himalayas. Based on results, morphological characters, micromorphological observations, qualitative and quantitative attributes, taxonomic keys for the studied taxa, distribution, and habitat are described. Stereoscope was used to analyze different qualitative characters, and light microscope was used to observe and measure the laminal and alar cells of the leaf. The SEM study reveals many important surface features like cell shape and cell wall. The laminal cells were mostly linear and elongated with thin cell walls. The specimens used for the SEM were air dried, so the laminal cells were somewhat constricted and concaved. The current study project will help to make the contribution in the taxonomy and morphology of this family.     

Presence of voids after warm vertical compaction and single‐cone obturation in band‐shaped isthmuses using micro‐computed tomography: A phantom study

The present micro‐computed tomography (micro‐CT) study compared the presence of voids in the band‐shaped isthmuses obturated by warm vertical compaction (WVC) and single‐cone (SC) techniques. Twenty mesial roots from mandibular first molar teeth showing a band‐shaped isthmus were selected and assigned into two groups based on their preoperative anatomical dimensions (n = 10), according to the filling technique: WVC or SC. Post‐filling micro‐CT scanning was performed. The percentage volume of root canal filling materials and voids were calculated and data were analyzed using the Mann–Whitney U test with a significance level of 5 and 95% confidence interval. Mean percentage volumes of filling materials and voids were 86.88 ± 8.53 and 13.11 ± 8.53 for the WVC group, 84.39 ± 8.30 and 15.60 ± 8.30 for the SC group, respectively, with no significant difference between them (p > .05). Neither WVC nor SC produced void‐free root canal fillings in the band‐shaped isthmuses and both techniques resulted in a similar quality of root canal filling.     

Establishing the archaeo‐metallurgic ornamentation process of an axe from the bronze age by OM,SEM‐EDX,and Micro‐FTIR

Our article presents the results of the analyses we performed by corroborating the Optical Microscopy, Scanning Electron Microscopy, Energy Dispersive Xray Analysis and micro Fourier Transformed InfraRed Analysis techniques to identify the archaeo‐metallurgic casting and ornamentation procedure of a decorated disk‐butted axe, which was discovered recently east of the Carpathian mountains, in the Moldavian Plateau. There are few known axes of that type found (A1, according to the usual typologies), as they are specific to the Middle Bronze Age period west of the Carpathians—the Wietenberg, Suciu de Sus, and Otomani‐Füzesabony cultures. The experimental data on the item under study revealed the fact that after casting it in molds made from porous silicone‐based stone, the object was coated with a thin layer, by immersing it in a lightly fusible tin alloy, whose main alloy component was copper and arsenic and iron as secondary components. After refining the shiny white layer, they applied a beautiful decoration pattern made by incision and engraving. This battle axe was an indication of higher status, such items usually being owned by community leaders. This important fact proves that the ancient metallurgic craftsmen were able to elaborate and manufacture various alloys from which they made beautiful objects and it also offers a new insight into the social and symbolic function of certain antique bronze items. Microsc. Res. Tech. 77:918–927, 2014. © 2014 Wiley Periodicals, Inc.     

A multimodal microcharacterisation of trace‐element zonation and crystallographic orientation in natural cassiterite by combining cathodoluminescence,EBSD, EPMA and contribution of confocal Raman‐in‐SEM imaging

In cassiterite, tin is associated with metals (titanium, niobium, tantalum, indium, tungsten, iron, manganese, mercury). Knowledge of mineral chemistry and trace‐element distribution is essential for: the understanding of ore formation, the exploration phase, the feasibility of ore treatment, and disposal/treatment of tailings after the exploitation phase. However, the availability of analytical methods make these characterisations difficult. We present a multitechnical approach to chemical and structural data that includes scanning electron microscopy (SEM)‐based imaging and microanalysis techniques such as: secondary and backscattered electrons, cathodoluminescence (CL), electron probe microanalyser (EPMA), electron backscattered diffraction (EBSD) and confocal Raman‐imaging integrated in a SEM (RISE). The presented results show the complementarity of the used analytical techniques. SEM, CL, EBSD, EPMA provide information from the interaction of an electron beam with minerals, leading to atomistic information about their composition, whereas RISE, Raman spectroscopy and imaging completes the studies with information about molecular vibrations, which are sensitive to structural modifications of the minerals. The correlation of Raman bands with the presence/absence of Nb, Ta, Fe (heterovalent substitution) and Ti (homovalent substitution) is established at a submicrometric scale. Combination of the different techniques makes it possible to establish a direct link between chemical and crystallographic data of cassiterite.     

Authentication of the 31 species of toxic and potent Chinese Materia medica (T/PCMM) by microscopic technique, part 1: three kinds of toxic and potent animal CMM

Microscopic authentication is an effective method for quality control of Chinese Materia Medica (CMM) because of its speed, convenience and low cost. However, the application of modern microscopic technique in quality evaluation of Toxic and Potent Chinese Materia Medica (T/PCMM) of animal origin is seldom reported. This gap in published knowledge is increasingly serious because confusion in T/PCMM has led to serious medical problems in China and other countries in recent years. To ensure the safe and effective use of T/PCMM, an accurate and convenient method, based on macroscopic and microscopic techniques, was developed for the authentication of animal T/PCMM. The color microscopic photos of the crude drug were acquired with the light microscope, and from these their morphological and microscopic characters were described. The present method was successfully applied in the analysis of 31 T/PCMM including 17 samples originating from plants, 3 from animals, 9 from minerals, and the remaining 2 from secreta. The macro- and microscopic characters of three animal T/PCMM, namely Mylabris (Mylabris phalerata or Mylabris cichorii), Huechys (Huechys sanguinea), and Lytta (Lytta caraganae) were conclusively determined and are here presented. The results demonstrated that it was feasible to use the present microscopic characters for the authentication of the above three animal T/PCMM.     

Tillandsia stricta Sol (Bromeliaceae) leaves as monitors of airborne particulate matter—A comparative SEM methods evaluation: Unveiling an accurate and odd HP‐SEM method

Airborne particulate matter (PM) has been included among the most important air pollutants by governmental environment agencies and academy researchers. The use of terrestrial plants for monitoring PM has been widely accepted, particularly when it is coupled with SEM/EDS. Herein, Tillandsia stricta leaves were used as monitors of PM, focusing on a comparative evaluation of Environmental SEM (ESEM) and High‐Pressure SEM (HPSEM). In addition, specimens air‐dried at formaldehyde atmosphere (AD/FA) were introduced as an SEM procedure. Hydrated specimen observation by ESEM was the best way to get information from T. stricta leaves. If any artifacts were introduced by AD/FA, they were indiscernible from those caused by CPD. Leaf anatomy was always well preserved. PM density was determined on adaxial and abaxial leaf epidermis for each of the SEM proceedings. When compared with ESEM, particle extraction varied from 0 to 20% in air‐dried leaves while 23–78% of particles deposited on leaves surfaces were extracted by CPD procedures. ESEM was obviously the best choice over other methods but morphological artifacts increased in function of operation time while HPSEM operation time was without limit. AD/FA avoided the shrinkage observed in the air‐dried leaves and particle extraction was low when compared with CPD. Structural and particle density results suggest AD/FA as an important methodological approach to air pollution biomonitoring that can be widely used in all electron microscopy labs. Otherwise, previous PM assessments using terrestrial plants as biomonitors and performed by conventional SEM could have underestimated airborne particulate matter concentration.     

Limit analysis of strain-hardening viscoplastic cylinders under internal pressure by using the velocity control: Analytical and numerical investigation

The paper aims to assess plastic limit loads of thick-walled hollow cylinders of strain-hardening viscoplastic materials under internal pressure. Particularly, the problem concerned features in the interaction between strengthening and weakening behavior during the deformation process. Therefore, the relating onset of instability and the stability condition also deserve to be further investigated. Analytical and finite-element limit analysis efforts are both made for complete and comparative investigation. By the concept of sequential limit analysis, the plastic limit loads were acquired by solving a sequence of limit analysis problems via computational optimization techniques. Applying the velocity control as a computational strategy to simulate the action of pressure, the paper investigates analytically and numerically the plastic limit load, the onset of instability and the stability condition of plane-strain circular cylinders. Especially, analytical solutions of the onset of instability were solved explicitly by the fixed point iteration. Validation of the present analytical and finite-element efforts was made completely with good agreement between the analytical solutions and the numerical results.     

Authentication of the 31 species of toxic and potent Chinese materia medica by microscopic technique assisted by ICP-MS analysis, part 4: four kinds of toxic and potent mineral arsenical CMMs

Toxic and Potent Chinese Materia Medica (T/PCMM) is a special and very important category of Chinese medicines. They have long been used in traditional medical practice and are being used more and more widely throughout the world in recent years. As there may be many fatal toxic effects caused by misusing or confusion of T/PCMM, their quality and safety control arouse increasing attention internationally. Researches on the accurate identification to ensure the safe use of T/PCMM are acquired; however, there are few reports on authentication. We are carrying out a series of studies on 31 T/PCMM originating from plants, animals, minerals, and secreta. In our previous studies, we proved that modern microscopic authentication is a simple, fast, effective, low cost, and less toxic method for identifying animal, seed, and flower T/PCMM. In the present study, we focused on the authentication of four kinds of mineral arsenicals, including orpiment (mainly containing As₂S₃), realgar (mainly containing As₄S₄), arsenolite, and arsenic trioxide (mainly containing As₂O₃). We examined the macroscopic and microscopic characteristics of the above minerals and found that they all can be easily identified and authenticated by using light microscopy coupled with polarized microscopy. Moreover, the authentication results for arsenolite and arsenic trioxide are confirmed by ICP‐MS analysis. We are sure that the morphological and microscopic characteristics indicated here are indispensable to establishing standards for these four mineral T/PCMMs. Microsc. Res. Tech. 74:1‐8, 2011. © 2010 Wiley‐Liss, Inc.     

Authentication of the 31 species of Toxic and Potent Chinese Materia Medica (T/PCMM) by microscopic technique, part 2: Three species of seed T/PCMM

Toxic and Potent Chinese Materia Medica (T/PCMM) are being used more and more in the treatment of various diseases. In view of their toxic side effects and to ensure their safe use, accurate and reliable authentication is indispensable. However, identifying characteristics of T/PCMM are seldom reported, even though modern microscopy can provide ample, unique identifying characteristics from cells found in transverse sections and powders. In particular, no systematic authentication studies on seed T/PCMM have been conducted. In the course of our study on 31 T/PCMM originating from plants, animals, minerals, and secreta, an accurate and convenient method, based on microscopic techniques, has been developed and reported for the authentication of animal T/PCMM. The present study deals with detailed investigations on three species of seed T/PCMM, namely Semen Hyoscyami (Hyoscyamus niger L.), Semen Euphorbiae (Euphorbia lathyris L.), and Semen Strychni (Strychnos nux-vomica L.). The macroscopic characters are here described in detail, and the microscopic characters were conclusively determined by common and polarized light microscopy. Results showed that these three T/PCMM can be easily identified by the present method even when powdered and combined. Thus, the microscopic method is applicable for authentication of the earlier three T/PCMM, and the morphological and microscopic characteristics described here are proposed as parameters to establish the authenticity of these three T/PCMM.     

Accurate and precise determination of isotopic ratios by MC‐ICP‐MS: A review

For many decades the accurate and precise determination of isotope ratios has remained a very strong interest to many researchers due to its important applications in earth, environmental, biological, archeological, and medical sciences. Traditionally, thermal ionization mass spectrometry (TIMS) has been the technique of choice for achieving the highest accuracy and precision. However, recent developments in multi‐collector inductively coupled plasma mass spectrometry (MC‐ICP‐MS) have brought a new dimension to this field. In addition to its simple and robust sample introduction, high sample throughput, and high mass resolution, the flat‐topped peaks generated by this technique provide for accurate and precise determination of isotope ratios with precision reaching 0.001%, comparable to that achieved with TIMS. These features, in combination with the ability of the ICP source to ionize nearly all elements in the periodic table, have resulted in an increased use of MC‐ICP‐MS for such measurements in various sample matrices. To determine accurate and precise isotope ratios with MC‐ICP‐MS, utmost care must be exercised during sample preparation, optimization of the instrument, and mass bias corrections. Unfortunately, there are inconsistencies and errors evident in many MC‐ICP‐MS publications, including errors in mass bias correction models. This review examines “state‐of‐the‐art” methodologies presented in the literature for achievement of precise and accurate determinations of isotope ratios by MC‐ICP‐MS. Some general rules for such accurate and precise measurements are suggested, and calculations of combined uncertainty of the data using a few common mass bias correction models are outlined. © 2009 Crown in the right of Canada. Published by Wiley Periodicals, Inc., Mass Spec Rev 28:990–1011, 2009