Samples Of Plant Material Research Articles

First European Erwinia amylovora Lytic Bacteriophage Cocktails Effective in the Host: Characterization and Prospects for Fire Blight Biocontrol.

Fire blight, caused by the plant-pathogenic bacterium Erwinia amylovora, is a highly contagious and difficult-to-control disease due to its efficient dissemination and survival and the scarcity of effective control methods. Copper and antibiotics are the most used treatments but pose environmental and human health risks. Bacteriophages (phages) constitute an ecological, safe, and sustainable fire blight control alternative. The goal of this study was to search for specific E. amylovora phages from plant material, soil, and water samples in Mediterranean environments. A collection of phages able to specifically infect and lyse E. amylovora strains was generated from former fire blight-affected orchards in Eastern Spain. Following in vitro characterization, assays in immature fruit revealed that preventively applying some of the phages or their combinations delayed the onset of fire blight symptoms and reduced the disease's severity, suggesting their biocontrol potential in Spain and other countries. The morphological and molecular characterization of the selected E. amylovora phages classified them as members of the class Caudoviricetes (former Myoviridae family) and genus Kolesnikvirus. This study reveals Mediterranean settings as plausible sources of E. amylovora-specific bacteriophages and provides the first effective European phage cocktails in plant material for the development of sustainable fire blight management measures.

ASSESSMENT OF THE POTENTIALLY TOXIC ELEMENTS CONTAMINATION AND THEIR UPTAKE BY AMBROSIA ARTEMISIIFOLIA L. IN SOILS IN THE NORTHERN PART OF BOSNIA AND HERZEGOVINA

This research focused on the determination of potentially toxic elements (PTEs) distribution in different agricultural soils and Ambrosia atremisiifolia L. (ragweed) at seven, different locations in the northern part of Bosnia and Herzegovina (BiH). Quantification of PTEs was done using atomic absorption spectrophotometry provided after acid digestion of the air-dried samples of soil and plant material. Determined content of elements in the soils increased as follows: Cd<Pb<Cu<Zn<Cr<Ni<Mn<Fe and were under the allowed maximum for unpolluted soils, except for Ni and Cr. However, transfer of Ni and Cr from the soil to the ragweed as well as their bioaccumulation was not intensive. Established metal contents ratio from plant tissues (root and shoot) to soil for both elements, represented their bioaccumulation factors (BAF), which were extremely low in the roots, as well as in the shoots, both in average less than 0.2. On the other hand, Zn showed a tendency to accumulate in ragweed shoots (BAF=2.07). The study showed that uptake and accumulation of PTEs in the ragweed was mainly influenced by their content in the soil, as well as specific characteristics and biological role of each element. Hence, content of PTEs in the ragweed tissue could be used as the soil contamination degree indicator.

Compliance Testing of Hemp (Cannabis sativa L.) Cultivars for Total Delta-9 THC and Total CBD Using Gas Chromatography with Flame Ionization Detection.

The United States Agriculture Improvement Act passed in December of 2018 legalized the growing of Cannabis sativa containing not more than 0.3% total Delta-9 tetrahydrocannabinol (THC) in the country. While Cannabis sativa has been cultivated for hundreds of years, the illegal status of the plant in the United States, and elsewhere, has hindered the development of plant cultivars that meet this legal definition. To assess sampling strategies, and conformance to the THC limit, 14 cultivars of hemp were grown and tested by using gas chromatography with flame ionization detection for total delta-9 THC and total cannabidiol (CBD) during 2020, 2021 and 2022. Each year, samples of fresh plant material were collected from each cultivar weekly, beginning in mid-August and ending in late October, to examine the rate of increase in THC and CBD for different cultivars and select individual plants. The sampling demonstrated that both CBD and THC increase rapidly over a 1-2-week time frame with maximum concentrations (about 16% and 0.6%, respectively) around late September to early October. The testing of individual plants on the same day for select cultivars showed that while the ratio of CBD to THC remains constant (about 20:1 in compliant hemp) during the growing season, the individual plants are highly variable in concentration. Whereas previous studies have shown cultivar-dependent variability in THC production, this study demonstrated a novel plant-to-plant variability in the levels of THC within the same hemp cultivar. Understanding variability within and between hemp cultivars is useful to determine field sampling strategies and to assess the risk of crop embargoes to growers by compliance regulators.

Chemical elements in Elaeis guineensis materials and derived oil

The production of oil palm (Elaeis guineensis) in Southeast Asia is vital to the economies of Indonesia and Malaysia. Both fertilisers and pesticides used in palm production can contain elevated concentrations of Trace Elements (TEs) which may accumulate in soils and leaf tissues of plants. We hypothesised that leaves from oil palms may be deficient in essential elements, while containing elevated concentrations of non-essential TEs commonly found in agrichemicals. Samples of plant materials (leaves and fruitlets) were collected from active and former plantations in Sumatra, Indonesia, and analysed for essential and non-essential elements. Indonesian palm oil samples were sourced in New Zealand and their elemental concentrations determined. Leaf materials from both active and abandoned production sites were deficient in N, K, S and Mo, while leaf materials from abandoned sites were deficient in P. These deficiencies may have been a contributing factor to the abandonment of production at these sites. Concentrations of non-essential elements were below or comparable to average plant concentrations and no evidence of contamination was found in plant tissues. Palm oil contained low concentrations of TEs, which did not pose any toxicity risks. However, Na and Al were present in concentrations of 1198 and 159 mg kg−1 respectively, which were higher than have been previously reported. Tropical oil palm production could benefit from the determination of bioaccumulation factors for fertiliser contaminants in E. guineensis, to limit the transfer of contaminants to plants and products if increased fertiliser applications were used to correct nutrient deficiencies.

Detection of saffron adulteration by other plant species using SPME-GC-HRMS

Saffron, a spice obtained by drying the scars of saffron flowers (Crocus sativus L.), is by far the most valuable spice in the world, and, not surprisingly, it often becomes an object of adulteration. In this study, non-target screening of volatile compounds in 38 authentic saffron samples and 25 samples of plant materials, potential saffron adulterants (safflower, calendula, capsicum and turmeric) was performed using headspace solid-phase microextraction for sample extraction followed by gas chromatography coupled to high-resolution mass spectrometry (HS-SPME-GC-HRMS) for non-target volatiles screening. The chemometric analysis of generated data by principal component analysis (PCA) and partial least squares discriminative analysis (PLS-DA) showed good separation of authentic saffron from potential plant adulterants. Significant compounds (‘markers’) for each group of these plants, as well as for saffron were tentatively identified. The target screening of selected ‘markers’ in model admixtures enabled simple and reliable detection levels as low as 2% w/w of safflower, calendula, capsicum or turmeric present in saffron.

Analysis of phenolic compounds in black walnut (Juglans nigra L.) fruit extract using high-performance liquid chromatography

Black walnut (Juglans nigra L.), which belongs to the Nut family (Juglandaceae), is a valuable raw material for the production of phytopreparations. The extract from the fruits of Juglans nigra L. has immunomodulating and antioxidant properties, which is primarily due to the presence of a large amount of phenolic compounds in the plant. But the phytochemical composition and pharmacological properties of Juglans nigra L. have not been sufficiently studied.
 The aim of our work was to study the optimal conditions for the extraction of Juglans nigra L. fruits, followed by the identification of its quantitative and qualitative composition.
 The main criterion for evaluating optimal extraction conditions was the quantitative content of phenolic compounds, which was determined spectrophotometrically by the Folin–Chocalteu method. The analysis of polyphenolic compounds was carried out by the method of high performance liquid chromatography.
 On the basis of experimental studies, the optimal conditions for extracting the fruits of Juglans nigra L. by maceration method conditions were selected. According to the results of the comparative analysis, it is shown that for the maximum extraction of phenolic compounds, it is advisable to use a 70% water-ethanol mixture as an extractant with a raw material to solvent ratio of 1:9. The method of high-performance liquid chromatography showed that the total content of phenolic compounds in the fruits of Juglans nigra L. is 2 327.09 μg/ml. Analysis of the extract of Juglans nigra L. by high-performance liquid chromatography method showed the presence of such phenolic compounds in plant material samples as catechins, catechin-like substances, flavonols (rutin, quercetin, myricetin), flavanones (naringin, naringenin) and phenolic acids. It should be noted, that the presence of such substances as chlorogenic acid, caffeic acid, kaempferol, hesperidin, genistein and daidzein glycosides, luteolin and apigenin glycosides, luteolin and apigenin in the extract of Juglans nigra L. was not detected.
 A high content of compounds of polyphenolic nature in the fruits of Juglans nigra L. was established. Therefore, this medicinal plant material is promising for further research, in particular, for the development of medicines based on the extract with potential anti-inflammatory, analgesic and antioxidant activity.

Antibacterial Activities of Extracts from four Wild Food Fruits

Aims: This study aimed to assess the in vitro antibacterial effects of extracts from four wild food fruits: Balanites aegyptiaca, Saba senegalensis, Ziziphus mauritiana, and Raphia sudanica.
 Place and Duration of Study: The samples of plant material were collected at Banamba and Sikasso, Mali between January and May 2018. The bacterial strains were collected at Research Centre for Biological Food and Nutritional Sciences (CRSBAN), University Professor Joseph Ki-Zerbo; Ouagadougou, Burkina Faso. The experimental parts were also carried out at CRSBAN from October 2019 to January 2020.
 Methodology: The fruit extracts were screened for antibacterial activity against Escherichia coli, Salmonella typhi, S. aureus, B. cereus, and L. monocytogenes strains. The diameters of the inhibition zones (ID), the minimum inhibitory concentrations (MIC) as well as the minimum bactericidal concentrations (MBC) were evaluated using agar diffusion method.
 Results: The findings revealed that these parameters have varied as a function of fruit species and/or their zones of provenances. All fruit extracts showed significant growth reducing effect against all the tested bacteria. The extracts from R. sudanica have exhibited the strongest growth-inhibiting activity specifically against E. coli (ID = 15.33±0.58 mm) and Salmonella typhi (ID = 18.00±1.00 mm) with lower MIC (from 2.08±1.44 to 5.83±1.44 mg/mL). Moreover, the MBC/MIC ratios revealed that the extracts from the studied fruits possess mainly bacteriostatic effects towards the tested strains.
 Conclusion: These findings support local therapeutics properties attributed to these fruits. They also demonstrate that, in addition to their nutritional values, these edible fruits could be used for developing antibiotics to treat infectious diseases and food poisoning.

STUDYING THE FEATURES OF ACCUMULATION OF FLAVONOIDS IN WORMWOOD HERB OF BITTER SYNANTROPIC FLORA OF THE ROSTOV REGION

Anthropogenic impact on medicinal plants is an important factor that affects the synthesis of secondary metabolites. With an increase in anthropogen-ic load, additional synthesis of secondary metabolites increases, primarily flavonoids, which play an important and obvious role in protecting plants from stress. The aim of the study was to study the features of the accumulation of flavonoids in the herb wormwood synanthropic flora of the Rostov region. Material and methods. Samples of medicinal plant material of wormwood herb collected in Morozovsky district of Rostov region were studied. Wormwood herb was collected and dried during the beginning of flowering (end of June - beginning of July 2021). The content of the sum of flavonoids in terms of rutin in selected samples of wormwood herb was determined three times by the standard pharmacopoeial method on a spectrophotometer SF-2000. Results. The study of vegetable raw materials harvested in the roadside zone of a slow road (IV category) with slow traffic and a railway showed a di-rect dependence on a decrease in the content of flavonoids with an increase in the distance from highways. Conclusion. According to the studies, it can be concluded that moderate anthropogenic impact, which is characterized by the release of various eco-toxicants into the environment, can induce the biosynthesis of flavonol compounds. However, under conditions of complex toxic stress (near the cate-gory IA highway with high transport traffic), the plant antioxidant system may be inhibited, which is expressed by the suppression of the production of polyphenols.

Etiology of Colletotrichum diseases on Sastuma mandarin in Croatia

During the last decade anthracnose has become a major disease of Satsuma mandarin, the most important citrus crop in Croatia. The aim of this study was to determine Colletotrichum species associated with different symptoms and to identify the origin of inoculum. From 2013 to 2016, 437 samples of plant material were collected. Colletotrichum spp. was isolated from 93% of dried twigs, 35% of dropped flowers, 89% of leaf spots, all fruit (100%) with anthracnose or calyx-end rot symptoms, 12% of fruit with post-harvest soft rot and from 40% of fruit showing spots remaining on trees after harvest. Out of 258 Colletotrichum isolates, 253 has been morphologically identified as C. gloeosporioides (Penz.) Penz. & Sacc. species complex. Twenty-seven representative isolates were selected for phylogenetic analysis. Sequencing the inter-spacer gene region of ribosomal DNA confirmed the identity of the species. Artificial inoculation of flowers led to more than 2-fold higher young fruit drop compared to control. Pathogenicity tests on green fruit induced typical anthracnose symptoms on 82% of inoculated fruit two months after inoculation. Inoculation of mature fruit caused the appearance of typical anthracnose symptoms on 87% of inoculated fruit. These results showed that C. gloeosporioides species complex is responsible for different disease types on Satsuma mandarin, and that the fungus is present throughout the year on different plant organs.

The Use of Flow Cytometry for Estimating Genome Sizes and DNA Ploidy Levels in Plants.

Flow cytometry has emerged as a uniquely flexible, accurate, and widely applicable technology for the analysis of plant cells. One of its most important applications centers on the measurement of nuclear DNA contents. This chapter describes the essential features of this measurement, outlining the overall methods and strategies, but going on to provide a wealth of technical details to ensure the most accurate and reproducible results. The chapter is aimed to be equally accessible to experienced plant cytometrists as well as those newly entering the field. Besides providing a step-by-step guide for estimating genome sizes and DNA-ploidy levels from fresh tissues, special attention is paid to the use of seeds and desiccated tissues for such purposes. Methodological aspects regarding field sampling, transport, and storage of plant material are also given in detail. Finally, troubleshooting information for the most common problems that may arise during the application of these methods is provided.

Decomposition of thiamethoxam in its combined use with hydroxycinnamic acids on potato and currant

The experiments were conducted in the city of Moscow, on the territory of the Educational, Scientific, and Production Center named after V. I. Edelstein “Michurinsky Garden” FSBEI HE “RSAU-Moscow Agricultural Academy named after K.A. Timiryazev” and Field experimental station in the period 2021-2022 on the black currant and potato plantings. Laboratory analysis of plant material samples for the study of the content of residual amounts and determination of the dynamics of pesticides destruction was carried out in the Educational, Scientific, and Consulting Center “Agroecology of pesticides and agrochemicals”. The following preparations were used in the tests: insecticide − Actara, VDG (250 g/kg of thiamethoxam) and growth regulator − Zircon, R (0.1 g/l of hydroxycinnamic acids). In the conditions of the experiments, a separate application of the preparation Actara, VDG and its combined use with the preparation Zircon, R to establish the level of residual amounts of the insecticide Actara, VDG active substance, sampling was carried out within 21 days from the moment of spraying on all experiment options. The selected samples were analyzed under laboratory conditions using the high-performance liquid chromatography method. As a result of the study, it was found that the use of Actar, VDG together with the preparation Zircon, R on black currant plantings caused an acceleration of the destruction of the insecticide active substance in the leaves. Nevertheless, the combined use of the studied preparations did not reduce the level of thiamethoxam intake in the forming black currant berries. The use of the preparation Actara, VDG together with the double use of the growth regulator Zircon, R on potato plantings led to accelerated decay of thiamethoxam in the tops and less of its accumulation in tubers.

Robust statistical analysis to predict and estimate the concentration of the cannabidiolic acid in Cannabis sativa L.: A comparative study

Growing awareness of the medicinal and therapeutic benefits of cannabis has resulted in extensive research, increasing numbers of commercial growers, and the provision of commercial testing. For commercial production, the importance of monitoring for consistent cannabinoid composition is essential. Gas chromatography and/or liquid chromatography methods are effective methods. Unfortunately, they are not easily scaled up for frequent or large-scale testing. Precision cultivation for medicinal cannabis requires the industry to test large volumes of plants frequently during different stages of the growth cycle. This cannot be achieved with destructive testing alone. A statistical method with 1H nuclear magnetic resonance (NMR) is proposed and investigated to explore its potential for a cost-effective mass-screening method of plant material samples. In addition, hyperspectral imaging (HSI) is employed for frequent non-destructive measurements. This allows increasing the frequency of observations during the growth of the plant. The cannabidiolic acid (CBDA) concentration is determined by the application of both HSI and NMR spectroscopy and validated against liquid chromatography-mass spectrometry (LCMS). The paper proposes a multivariate statistical regression algorithm that automatically determines the CBDA concentration directly from bucket integration of the NMR spectrum. The algorithm successfully predicted the CBDA concentration of 7 unknown samples from data selected from 4 known samples, with an average R2 value of 0.98. The proposed statistical method was applied to the data collected for HSI. It showed that while the hyperspectral dataset can be correlated with CBDA concentration, it was subject to high variance. However, HSI retains the spatial information of the actual plant structure, allowing the CBDA prediction to be mapped back to the original spatial location in the plant while providing visual information on CBDA concentration within a flower or leaf without requiring the plant component to be destroyed in the process.

Characterization of Fusarium spp. causal agent of wilt in pecan

One of the limiting factors for pecan production is the incidence of disease caused by Fusarium spp. which are a threat to orchards in the south of the country. Therefore, the study aimed to confirm the pathogenicity and evaluate the morphophysiological and molecular characteristics of Fusarium spp. associated with pecan in Rio Grande do Sul and Paraná. For this, samples of symptomatic plant material were collected, and potentially pathogenic isolates were evaluated for pathogenicity, by immersing the roots of the plants in a spore suspension of Fusarium spp. In addition, the severity of the disease was assessed using a rating scale according to the symptoms expressed by the plants. The isolates were also analyzed for morphophysiological variability, through evaluations of mycelial growth, colony and aerial mycelium pigmentation, and characteristics of reproductive structures. Molecular characterization was performed which amplify the region of the 1-alpha elongation factor and sequencing. Nineteen isolates were obtained, which were considered pathogenic, however, there was variability in their aggressiveness and morphophysiological characteristics. In addition, the sequencing was used to identify different species. Therefore, species of Fusarium are responsible for the occurrence of wilt in pecan in southern Brazil.

Bauhinia forficata Link Infusions: Chemical and Bioactivity of Volatile and Non-Volatile Fractions.

This study aimed to evaluate Bauhinia forficata infusions prepared using samples available in Rio de Janeiro, Brazil. As such, infusions at 5% (w/v) of different brands and batches commercialized in the city (CS1, CS2, CS3, and CS4) and samples of plant material botanically identified (BS) were evaluated to determine their total phenolic and flavonoid contents (TPC and TFC), antioxidant capacity (ABTS•+, DPPH•, and FRAP assays), phytochemical profile, volatile compounds, and inhibitory effects against the α-amylase enzyme. The results showed that infusions prepared using BS samples had lower TPC, TFC and antioxidant potential than the commercial samples (p < 0.05). The batch averages presented high standard deviations mainly for the commercial samples, corroborating sample heterogeneity. Sample volatile fractions were mainly composed of terpenes (40 compounds identified). In the non-volatile fraction, 20 compounds were identified, with emphasis on the CS3 sample, which comprised most of the compounds, mainly flavonoid derivatives. PCA analysis demonstrated more chemical diversity in non-volatile than volatile compounds. The samples also inhibited the α-amylase enzyme (IC50 value: 0.235–0.801 mg RE/mL). Despite the differences observed in this work, B. forficata is recognized as a source of bioactive compounds that can increase the intake of antioxidant compounds by the population.

Quantitative evaluation of the composition of synthetic cannabinoid receptor agonists (SCRA) in infused papers and herbal materials seized in Brazilian prisons

This study describes the identification and quantification of synthetic cannabinoid receptor agonists (SCRA) in infused paper and herbal material samples, seized in prisons located in São Paulo state, Brazil. The selected samples (23 infused paper and 3 herbal material samples) were previously analyzed by the São Paulo State Police, in a qualitative analysis by gas chromatography–mass spectrometry (GC–MS) using full-scan and library search for identification. Afterward, these samples were analyzed by liquid chromatography-tandem mass spectrometry in a triple quadrupole (LC-MS/MS) system operating in MRM mode for determination of the majority and the possible minority SCRA present in the samples. For the LC-MS/MS analysis the samples were extracted with methanol and diluted in a proportion of 1:1000 and 1:10,000. All substances identified by GC-MS were also identified by LC-MS/MS, but not the opposite, showing GC-MS full-scan method employed were unable to detect those SCRA presented in lower concentration levels. The presence of a single SCRA in the samples was observed in 11 samples (27.5%), while in 29 samples (72.5%) two or more SCRA were identified. The SCRA most frequently detected in the samples were MDMB-4en-PINACA and ADB-BUTINACA, present in 72% and 70% of the samples, respectively. The concentration of SCRA shown great variability, ranging from 0.2 μg/infused paper to 3000 μg/infused paper. In cases with more than one SCRA detected, the minority compounds did not exceed the concentration of 50 μg/infused paper. Comparing two different samples of a single seizure it was possible to identify the same substances but in different concentration levels, or different compositions (mixtures). The highest concentration of SCRA present in the infused papers was for the cases involving MDMB-4en-PINACA as the major compound, reaching until 3000 μg/infused paper, but this SCRA was also found as minority in other cases, with concentrations as low as 0.9 μg/infused paper. Among all samples, 4F-MDMB-BUTICA was detected in only one case, at a concentration of 125 μg/infused paper. For the three samples of plant material, ADB-BUTINACA appears as the major compound, with the presence of other minor SCRA ranging from 0.2 to 5 μg/mg. These results shows that the studied samples did not present homogeneity in the composition and concentration levels, even when comparing a single case, but it seems that some SCRA are commonly found together. The presence of different SCRA in the same sample may contribute to the increase in the toxicity of these compounds, which reinforces the need to obtain quantitative and qualitative data from the seized samples.

Greenness of procedures using NADES in the preparation of vegetal samples: Comparison of five green metrics

The principles of green analytical chemistry have led to the development of analytical procedures that are increasingly sustainable. Different metrics have been created for the evaluation of greenness, although determination of the green nature of new analytical methods remains challenging, including for extraction methods that involve the use of natural deep eutectic solvents (NADES). In this study, the following five chemical metrics for the evaluation of greenness were considered: National Environmental Methods Index (NEMI), Green Analytical Procedure Index (GAPI), Analytical Eco-Scale, Analytical GREEnness (AGREE), and White Analytical Chemistry (WAC). These methods were applied in evaluation of the environmental and sustainability characteristics of three different methods for the preparation of plant material samples: microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and microwave-assisted acid digestion (MW-AD). These methods employed different NADES as extraction solvents, as well as dilute nitric acid as an oxidizing agent, for the determination of As, Cd, Pb, and V by inductively coupled plasma mass spectrometry (ICP-MS). The NEMI metric found no differences between the MAE-NADES and UAE-NADES methods. The GAPI metric found differences between the MAE-NADES and UAE-NADES methods and identify the disadvantageous aspects of each step of the methods. The Analytical Eco-Scale and AGREE identified the MAE-NADES method as the greenest, while WAC-12 RGB indicated the UAE-NADES method as the greenest procedure. A detailed discussion is provided of the application of each metric, together with their differences and advantages.

Carbon from above-ground biomass and litter accumulated in an Atlantic Forest fragment

This work aimed to quantify carbon (C) stocks from above-ground biomass (AGB) and accumulated litter in a fragment of the Atlantic Forest biome (edge and interior) in Northeastern Brazil. The study was carried out in areas with native vegetation, in a fragment of Atlantic tropical forest, municipality of Sirinhaém, Pernambuco, Brazil. AGB was quantified using the allometric equation proposed for tropical rainforests. The samples of plant material were dried at 60°C, weighed and ground, and then taken to the laboratory for analysis of organic C contents by dry combustion. AGB stocks were not different between the fragment edge and interior environments. The AGB stratification into diameter classes showed that large trees are important for the storage of biomass in trees. Litter biomass stocks also did not differ between edge and interior. The branch compartment showed higher levels of C. The C-AGB stocks were not different between edge and interior, similar to the C-litter. C stocks did not differ between interior and edge among tree species, except for Protium heptaphyllum, which had the highest C stocks at edges. With this study, we hope to contribute to improving C estimates in fragmented forests to support forest conservation policies, support REDD+ projects, and access to carbon credits

Potency testing of up to twenty cannabinoids by liquid chromatography diode array detector with optional electrospray ionization time-of-flight mass spectrometry

The growing popularity of Cannabis sativa L. and its widespread use for medical and recreational purposes have created an urgent demand of accurate and reliable analytical methods to identify and quantify a growing number of cannabinoids. To meet this demand, a liquid chromatography diode array detector (LC-DAD) method has been developed, validated, and applied in analysis of cannabinoids in nine samples of plant materials of marijuana, six samples of marijuana cigarettes, five samples of hemp flowers, one sample of hemp cigarette, and two samples of Δ8-tetrahydrocannabinol (Δ8-THC) fortified hemp flowers. The method has achieved significant improvements over published methods, which was characterized of 20 targeted cannabinoids, 18 quantified cannabinoids, baseline resolution among quantified cannabinoids, a low limit of quantification (0.02 μg mL−1), a wide linear range (0.02–25 μg mL−1 or 0.04–50% (w/w)), a unique experiment to track the recovery of sample preparation in real time by spiking abnormal cannabidiol (CBD), and a well-assessed specificity by electrospray ionization time-of-flight mass spectrometry (ESI/TOFMS). Precision and accuracy were assessed using quality control (QC) samples at three concentration levels, i.e., 0.02, 0.5, and 12.5 μg mL−1, in triplicates with inter-day and intra-day precision of less than 15% relative standard deviation (RSD) and accuracy of less than ±15% relative error, therefore meeting the requirements by the ISO 17025 standards. Additionally, ESI/TOFMS has discovered seven unknown cannabinoids, including one structural isomer of cannabigerol (CBG), one structural isomer of cannabinolic acid (CBNA), four structural isomers of Δ9-THC, and one structural isomer of Δ9-THC acetate. Furthermore, it uncovered that one of the two samples of Δ8-THC fortified hemp flowers contained 5.16% (w/w) Δ9-THC, an alarm to the current Δ8-THC craze by the public.

Simultaneous Determination of Multi-Class Pesticide Residues and PAHs in Plant Material and Soil Samples Using the Optimized QuEChERS Method and Tandem Mass Spectrometry Analysis.

New analytical approaches to the simultaneous identification and quantification of 94 pesticides and 13 polycyclic aromatic hydrocarbons (PAHs) in five representative matrices (pepper, apple, lettuce, wheat, and soil) were developed. The analyses were based on gas chromatography coupled with triple quadrupole tandem mass spectrometry (GC-MS/MS). The procedure was optimized by changing the solvent used during the extraction, from acetonitrile to the acetone: n-hexane mixture at a volume ratio of 1:4 (v/v), as well as the use of a reduced amount of water during the extraction of compounds from cereals. An additional modification was the use of florisil instead of GCB in the sample cleanup step. A full method validation study was performed, at two concentration levels (LOQ and 1000 × LOQ), which showed satisfactory results for all analytes from the PAHs group, with recoveries ranging from 70.7–115.1%, and an average RSD of 3.9%. Linearity was tested in the range of 0.001–1.000 mg/kg and showed coefficients of determination (R2) ≥ 0.99 for all PAHs. Satisfactory recovery and precision parameters (LOQ and 100 × LOQ) were achieved for almost all analytes from the pesticide group in the range of 70.1–119.3% with the mean RSD equal to 5.9%. The observed linearity for all analytes in the concentration range of 0.005–1.44 mg/kg was R2 ≥ 0.99, with the exception of famoxadone, chizalofop-p-ethyl, prothioconazole, spirodiclofen, tefluthrin, and zoxamid. The extended uncertainties were estimated, using a top-down approach of 9.9% (average) and 15.3% (average) for PAHs and pesticides samples, respectively (the coverage factor k = 2, the 95% confidence level). Ultimately, the method was successfully applied to determine pesticide residues in commercial samples of fruit, vegetables and grain, and soil samples for PAHs, which were collected from selected places in the Podkarpacie region. A total of 38 real samples were tested, in which 10 pesticides and 13 PAHs were determined. Proposed changes allow us to shorten the sample preparation time (by 20%) and to reduce the consumption of organic solvents (by 17%). The use of florisil for sample cleanup, instead of GCB, improves the recovery of compounds with flat particles.

Can plants indicate where a corpse is buried? Effects of buried animal tissues on plant chemistry: Preliminary study

The bodies of murder or accident victims may be concealed below ground to cover up the crime by the perpetrator. During decomposition, nutrient-rich substances that may affect the surrounding environment enter the soil. The logical effect of local changes in soil biogeochemistry should be an impact on the vegetation occurring in a cadaver decomposition island (CDI). Nitrogen, phosphorus, and potassium compounds, which are common plant fertilizers, have received most of the attention in studies using animal remains, but many other elements such as trace metals enter the environment during decomposition of carcass. Furthermore, the decomposition of carcasses can locally affect environmental parameters such as humidity, soil pH, and oxygenation. This study examined whether animal tissues buried in soil could affect vegetation. The experiment was conducted in an open space in southern Poland, using limbs from Sus scrofa domestica. Four test plots were set up: I – clean soil (control plot), II – control plot + urine (potential nitrogen source), III – animal tissues buried at 20 cm depth and IV – animal tissues buried at 10 cm depth. Seedlings of Begonia semperflorens were planted in all four plots. After the end of one full growing season, plant measurements were obtained, and soil and plant material samples were taken. The presence of limbs, regardless of depth, had a very positive effect on plant development. Chemical analyses were performed to determine which compounds could have affected the vegetation. In addition to the standard analyses for nitrogen, phosphorus or potassium, levels of copper, iron, manganese, zinc and magnesium were also determined. The elemental content in the soil and plant material from different plots varied. Interestingly, despite the more intense vegetation, lower levels of elements were found in the soil and leaves of plants from the experimental than from the control plots. In many cases, there were clear differences in the contents of elements between the experimental plots. This could indicate, for example, different phases of animal tissue decomposition, which could have been caused by the different depths at which the remains were buried. As concentrations of elements in soil and plants can change significantly over time, material will need to be collected at regular intervals in any future studies. However, it is still unknown what exactly is responsible for the enhanced plant growth within a CDI and whether there are markers that could be helpful in identifying the burial sites of carcasses on the basis of comprehensive vegetation analysis.