Liquid Chromatography Fractionation Research Articles

Purification of mineral oil aromatic hydrocarbons and separation based on the number of aromatic rings using a liquid chromatography silica column. An alternative to epoxidation.

The analysis of mineral oil aromatic hydrocarbons (MOAH) in vegetable oils is currently associated with high uncertainty due to various factors ranging from sample preparation to data interpretation. One significant factor is the coelution of biogenic compounds of terpenic origin with the MOAH fraction during chromatographic analysis. The common purification method is epoxidation, a chemical reaction that changes the polarity of the interferences, allowing their separation from MOAH. However, this reaction is non-selective and can lead to losses of both MOAH and internal standards. This variation is especially noticeable when using epoxidation procedures with higher reaction kinetics, such as those employing performic acid. MOAH losses also vary depending on their composition, which is unpredictable. Furthermore, 2-methylnaphthalene (2MN) and 1,3,5-tri‑tert-butylbenzene (TBB), which are the common quantification standards, are lost at different rates. As a consequence, the final MOAH quantity will vary both depending on the standard used, and on its initial composition. This work presents a new purification approach based on liquid chromatography fractionation on silica using the same column and eluents as for the usual MOSH/MOAH fractionation. This approach efficiently removes squalene, carotenes, and derivatives, without inducing inconsistent losses between the internal standards and MOAH as epoxidation does. On average, MOAH recovery using this new method was 94% (± 8%) in coconut, palm, sunflower, and olive oil, using different MOAH sources and concentrations. Additionally, perspectives are presented regarding the separate analysis and quantification of mono-/diaromatic MOAH and other polyaromatic MOAH. This is of particular interest as these two sub-fractions are associated with different toxicological properties.

Chiral supercritical fluid chromatography-mass spectrometry with liquid chromatography fractionation for the characterization of enantiomeric composition of fatty acid esters of hydroxy fatty acids

Chiral supercritical fluid chromatography-mass spectrometry with liquid chromatography fractionation for the characterization of enantiomeric composition of fatty acid esters of hydroxy fatty acids

Macrophage foam cell-derived mediator promotes spontaneous fat lipolysis in atherosclerosis models.

Ectopic lipid accumulation in macrophages is responsible for the formation of macrophage foam cells (MFCs) which are involved in the crosstalk with the perivascular adipose tissue (PVAT) of the vascular wall that plays a pivotal role in the progression of atherosclerosis. However, the interrelationship between MFCs and PVAT implementing adipocyte dysfunction during atherosclerosis has not yet been established. We hypothesized that MFC-secreted mediator(s) is causally linked with PVAT dysfunction and the succession of atherosclerosis. To test this hypothesis, MFCs were cocultured with adipocytes, or the conditional media of MFCs (MFC-CM) were exposed to adipocytes and found a significant induction of fat lipolysis in adipocytes. The molecular filtration followed by the high-performance liquid chromatography (HPLC) fractionation and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis of MFC-CM revealed a novel mediator fetuin-A (FetA) that significantly augments toll-like receptor 4 (TLR4)-dependent fat lipolysis in adipocytes. Mechanistically, MFC-derived FetA markedly increased TLR4-dependent c-Jun N-terminal kinases (JNK)/extracellular signal-regulated kinases (ERK) activation that causes spontaneous fat lipolysis implementing adipocyte dysfunction. Thus, the present study provides the first evidence of MFC-derived FetA that induces adipocyte dysfunction by the stimulation of spontaneous fat lipolysis. Therefore, targeting the crosstalk between MFCs and adipocytes could be a newer approach to counter the progression of atherosclerosis.

Aktivitas Antioksidan Fraksi Larut Etil Asetat dari Ekstrak Akar Tabar Kedayan (Aristolochia papilifolia Ding Hou)

Kedayan root is a medicinal plant that is traditionally used by the community as medicine, especially as an antidote to poison. This research aimed to determine the antioxidant activity of the ethyl acetate soluble fraction of Tabar Kedayan root extract (Aristolochia papillfolia Ding hou). The fractionation method used is solid-liquid extraction using ethyl acetate solvent to obtain a soluble fraction of ethyl acetate, then vacuum liquid chromatography fractionation using the eluent ratio n-hexane: ethyl acetate and obtain fraction A, fraction B, fraction C, fraction D, fraction E, fraction F. Testing of antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method spectrophotometrically. The data obtained was analyzed using linear regression to determine IC50. The results of the C fraction test on the ethyl acetate soluble fraction of Tabar Kedayan roots (A. papillfolia) have antioxidant activity with an IC50 value of 139.11 ppm.

Exploiting Soybean and Flaxseed Meal Byproducts as Safe Weed Management Approaches in Onion Field

The intensive and repeated use of chemical herbicides has led to the emergence of herbicide-resistant weeds, which, in addition to their environmental impacts, also pose significant threats to human and animal health. This study aimed to explore the potential of oilseed industrial wastes, specifically soybean and flaxseed meals, as safe and environmentally friendly bioherbicides for controlling weeds associated with onion crops. Two field experiments were conducted along two successive winter of 2020/21 and 2021/22. Treatments involved foliar spray of soybean and flaxseed meals in three different concentrations (15, 30 and 45%), mulching of seed meals, oxyfluorfen herbicide, two hand hoeing and untreated weedy check. The findings demonstrated that all weed control treatments significantly reduced weed density, biomass and nutrient uptake. Two hand hoeing, oxyfluorfen herbicide and the mulching of soybean and flaxseed meals alternated in the top rank for weed control, showing minimal significant differences among them. Following these effective treatments, soybean meal extracts at 45 and 30% exhibited notable weed control compared to the weedy check. The greatest enhancement in onion growth, yield characteristics and bulb quality was observed with the application of hand hoeing, soybean and flaxseed meal mulching treatments, with no significant differences between them. High-Performance Liquid Chromatography (HPLC) fractionation of both meals identified various phenolic acids at different concentrations. Practically, these safe efficient treatments proved progress on chemical herbicide. Hence, onion farmers are advised to apply soybean and flaxseed meals mulching safe treatments as alternative to harmful chemical herbicides under all experimental conditions.

Evaluation of Antimicrobial Activity and Phytoconstituents of Stem Bark Extract and Vacuum Liquid Chromatographic Fractions of Spondias mombin (Linn)

Evaluation of Antimicrobial Activity and Phytoconstituents of Stem Bark Extract and Vacuum Liquid Chromatographic Fractions of Spondias mombin (Linn)

Cytotoxic Potential of Compounds Isolated from Non-Polar Fractions of Sungkai Plant Leaves (Peronema canescens Jack) Against Artemia salina Leach Larvae

The sungkai plant (Peronema canescens Jack), belonging to the Lamiaceae family, is a plant that is traditionally used as medicine, including toothache, malaria, and fever medicine. In this research, isolation was carried out with vacuum liquid chromatography (VLC), solid and liquid fractions were obtained. The solid fraction was further separated using column chromatography to obtain the isolated compound as a white solid (amorphous) weighing 10 mg (melting point 140˚C-142˚C). The results of the UV spectrum data show that there are no conjugated double bonds. The results of the IR spectrum show the presence of C-H groups at wave numbers 2921,49 cm-1 and 2856,94 cm-1, C=O groups at wave numbers, C=C groups at wave numbers 1641,51 cm-1, and dimethyl germinal which is characteristic of triterpenoid compounds at wave numbers 1456.68 cm-1 and 1372.41 cm-1. Meanwhile, the isolated oil was analyzed for chemical components using GC-MS. It was discovered that there were 83 chemical compound components contained therein with 4 main compound components, namely pentadecanoic acid (16.65%), 9,12-octadecanoic acid (16.12%), propyl palmitate (7.89%), and hexadecenoic acid, methyl ester (5.59%). A cytotoxic test was carried out on both fractions using the Brine Shrimp Lethality Test (BSLT) method. The results showed that the isolated compound was non-toxic with an LC50 value of 190214.2807 mg/L and the isolated oil was very toxic with an LC50 value of 34.2452 mg/L.

Liquid chromatographic fractionation of bio-oil from sugarcane bagasse: influence of heating rate on bio-oil yield and quality

Brazil stands out in the field of using biomass as a source of energy and biomaterials, due to its territorial extension, biodiversity and climatic conditions. In this sense, one can highlight the high potential of bio-products generated from biomass. This is the case of sugarcane bagasse, produced in large quantities allied to the sugar and alcohol industry. In Brazil, much research has been done to improve its amount of fiber, aiming at the production of alternative fuels and generating the so-called "energy cane". Thus, it was studied the use of sugarcane bagasse for the production of bio-oil through pyrolysis and the isolation of fractions for industrial application. A comparison was also made between two types of sugarcane, a commercial variety (Saccharum sp.) and a variety with some genetic improvement (Erianthus arundinaceus). The final pyrolysis temperature was set at 500 ° C by varying the heating rates (25, 45, and 65 ° C min-1). The bio-oils were fractionated using preparative liquid chromatography and their fractions were analyzed by gas chromatography coupled to mass spectrometry. The fractionation of bio-oils increased the number of compounds identified by about 50%, besides allowing the isolation of apolar compounds. In addition, it was found that the genetic improved sugarcane presented higher bio-oil content, with higher hydrocarbon content, when compared to commercial sugarcane, demonstrating that the improvement process was efficient. Among the compounds identified were phenols, furfural derivatives and hydrocarbons, which indicates the potential use of bio-oil not only as bio-fuels, but also for industrial purposes.

Optimized Lipidomics Extraction of Sphingosine and Sphinganine from Optic Nerve for Signaling Studies.

Interconvertible sphingolipid metabolites represent germane constituents of eukaryotic membranes and are vital in the regulation of cellular homeostasis, proliferation, survival, and induction of autophagy. This protocol describes a step-by-step method for extractions of sphingosine and sphinganine from mammalian tissue samples, particularly from the murine optic nerve. These lipids are partitioned into a binary mixture of chloroform and methanol in a modified Bligh and Dyer method. This is followed with reverse phase ultrahigh-performance liquid chromatography fractionation with a C18+ column and subsequent tandem mass spectrometry (UHPLC-MS-MS) analysis of the biological abundance. These free sphingoid bases dissociate to form structurally distinctive carbocation product ions that can be confirmed with annotations of lipidomic databases or in-house fragmentation software.

Bile acids as putative social signals in Mozambique tilapia (Oreochromis mossambicus)

Chemical cues provide potential mates with information about reproductive status and resource-holding potential. In the Mozambique tilapia (Oreochromis mossambicus), males can distinguish female reproductive status through chemical cues, and accessibility of males to females depends on their position in the hierarchy, determined in part by chemical cues. Here, we hypothesized that tilapia faecal cues are attractive to conspecifics once released into the water. C18 solid-phase extracts of faeces from dominant males and pre-ovulatory females evoked stronger olfactory epithelium electrical responses (EOG) than, respectively, subordinate males and post-spawning females. Mass spectrometry of the reverse-phase C18 high-performance liquid chromatography fractions of these extracts with highest EOG, identified by amino acids and bile acids. Faeces from pre-ovulatory females contain significantly higher concentrations of cholic acid (CA) and taurocholic acid (TCH) than both post-spawning females and males. A pool of amino acids had no effect on aggression or attraction in males. However, males were attracted to the scent of pre-ovulatory female faeces, as well as CA and TCH, when applied separately. This attraction was accompanied by increased digging behaviour compared to the odour of post-spawning females. CA and TCH exert their action through separate receptor mechanisms. These findings are consistent with a role for faeces – and bile acids therein – in chemical communication in this species, acting as an attractant for males to reproductive females.

Large-Scale Identification of Lysine Crotonylation Reveals Its Potential Role in Oral Squamous Cell Carcinoma.

Lysine crotonylation, an emerging posttranslational modification, has been implicated in the regulation of diverse biological processes. However, its involvement in oral squamous cell carcinoma (OSCC) remains elusive. This study aims to reveal the global crotonylome in OSCC under hypoxic conditions and explore the potential regulatory mechanism of crotonylation in OSCC. Liquid-chromatography fractionation, affinity enrichment of crotonylated peptides, and high-resolution mass spectrometry were employed to detect differential crotonylation in CAL27 cells cultured under hypoxia. The obtained data were further subjected to bioinformatics analysis to uncover the involved biological processes and pathways of the dysregulated crotonylated proteins. A site-mutated plasmid was utilized to investigate the effect of crotonylation on Heat Shock Protein 90 Alpha Family Class B Member 1 (HAP90AB1) function. A large-scale crotonylome analysis revealed 1563 crotonylated modification sites on 605 proteins in CAL27 cells under hypoxia. Bioinformatics analysis revealed a significant decrease in histone crotonylation levels, while up-regulated crotonylated proteins were mainly concentrated in non-histone proteins. Notably, glycolysis-related proteins exhibited prominent up-regulation among the identified crotonylated proteins, with HSP90AB1 displaying the most significant changes. Subsequent experimental findings confirmed that mutating lysine 265 of HSP90AB1 into a silent arginine impaired its function in promoting glycolysis. Our study provides insights into the crotonylation modification of proteins in OSCC under hypoxic conditions and elucidates the associated biological processes and pathways. Crotonylation of HSP90AB1 in hypoxic conditions may enhance the glycolysis regulation ability in OSCC, offering novel perspectives on the regulatory mechanism of crotonylation in hypoxic OSCC and potential therapeutic targets for OSCC treatment.

Determining factors of durum wheat bread loaf volume and alveograph characteristics under optimal, drought and heat stress conditions

Durum wheat is an important crop, especially in Mediterrannean countries; around 25% of durum wheat world-wide, and 70–90% in the Middle East, is used for bread making. Climate change is causing increased periods of drought stress and higher temperatures which are affecting wheat quality. Alveograph extensibility is important for durum wheat loaf volume (LFV), as is the relationship between dough tenacity and extensibility. This study was done to identify the factors determining LFV and alveograph properties under optimal and stress conditions. Six durum wheat cultivars with the same glutenin composition were grown for two seasons under optimal, drought and heat stress conditions. Good LFV was consistently related with higher dough extensibility, irrespective of production conditions. A number of the LMW glutenin fractions, and the γ-gliadins were significantly reduced and β-gliadins were increased under both heat and drought stress. The large and total unextractable polymeric proteins and unextractable HMW glutenins were significantly reduced by heat stress. In a multiple regression of LFV (without alveograph values), solvent retention capacity and sodium dodecyl sulphate sedimentation were the best predictors of LFV under all conditions. Size-exclusion high-performance liquid chromatography fractions combined with flour protein content were good predictors of LFV only under optimal conditions.

Synergistic enhancement of glucagon-like peptide-1 release by γ-aminobutyric acid and L-phenylalanine in enteroendocrine cells-searching active ingredients in a water extract of corn zein protein.

This study investigated the glucagon-like peptide-1 (GLP-1)-releasing activity of an aqueous extract (ZeinS) from corn zein protein and aimed to identify the active compounds responsible for this activity. Glucagon-like peptide-1-releasing activity was evaluated using a murine enteroendocrine cell line (GLUTag). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on purified fractions of ZeinS to identify active molecules. ZeinS stimulated more GLP-1 secretion from GLUTag cells compared to zein hydrolysate. Fractions displaying biological activity were determined by solid-phase extraction and high-performance liquid chromatography (HPLC) fractionation. Subsequent LC-MS/MS analysis identified several amino acids in the active fractions of ZeinS. In particular, γ-aminobutyric acid (GABA) exhibited significant GLP-1-releasing activity both alone and synergistically with L-phenylalanine (Phe). Moreover, ZeinS-induced GLP-1 secretion was attenuated by antagonists for the GABA receptor and calcium sensing receptor. These results demonstrate that GABA and Phe identified in ZeinS synergistically stimulate GLP-1 secretion in enteroendocrine cells.

Ultra-high-performance liquid chromatography-tandem high-resolution elevated mass spectrometry profiling of anti-methicillin-resistant Staphylococcus aureus metabolites from the endophytic bacteria collected from the weeds of a previous dumpsite

The culturable endophytic bacteria from the weeds Cleome rutidosperma of the family Cleomaceae and Digitaria sanguinalis of the family Poaceae obtained from a previous dumpsite in Pampanga, Philippines have been assessed for their anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and the analytes with such activity should be identified. However, due to the limited amounts collected from the isolation process, 1.8 mg yield of compound 1 from the endophyte of C. rutidosperma and 1.2 mg of a mixture from the endophyte of D. sanguinalis were selected for LC-MSE analysis. The production of compounds from the culturable endophytic bacteria Pseudomonas aeruginosa— determined by gene-sequencing, an untargeted and data-independent analysis (DIA) by ultra-high performance liquid chromatography-high resolution-elevated energy mass spectrometry (UHPLC-HR-MSE) technique was employed to profile the metabolites present in the two high-performance liquid chromatography (HPLC) fractions. The analytes present from P. aeruginosa detected by UHPLC-HR-MSE isolated from C. rutidosperma was phenazine-1-carboxylic acid (1), and for D. sanguinalis were chamigrenal (2), dialkyl resorcinol (3), and a pyoverdine elicitor (4). This study proves that UHPLC-HR-MSE could identify the anti-MRSA constituents in P. aeruginosa from commensal weeds C. rutidosperma and D. sanguinalis. The UHPLC-HR-MSE could help strengthen metabolomics antibacterial research and its related applications from a future perspective. Application of metabolomics research using UHPLC-HR-MSE could enhance the rehabilitation of dumpsites by the microbial community present.

Screening of secondary metabolites produced by a mangroove-derived Nigrospora species an endophytic fungus isolated from Rhizophora racemosa for antioxidant and antimicrobial properties

Marine fungi have shown large chemo-diversity of unharnessed important secondary metabolites required for drug development. In this study, endophytic fungi Nigrospora species (5S3) was isolated from the fresh stem of Rhizophora racemosa from a mangrove forest in Lagos Nigeria and axenic colonies were fermented on sterile rice medium for 21 days. The antimicrobial and antioxidant activities of the extract were evaluated. Chemical analyses such as High Performance Liquid Chromatography Diode Array Detector (HPLC-DAD) and Nuclear magnetic Resonance Imaging (NMR) analyses of selected Vacuum Liquid Chromatography (VLC) fractions revealed the presence of ten known compounds with established biological activities. Tested fractions of the extract exhibited antimicrobial activities against at least one Gram positive and Gram negative bacteria with MIC values that ranged between 0.13 to 1 mg/mL. FE2 and FE3 fractions of the extract demonstrated poor antioxidant activity of less than 10%. Septicine, Aureonitol, Papuamine, Di-iso-octylphtalat, 1-(2,4 Dihydroxy-3,5-Dimethylphenyl)-Ethanone, Cladosporin, Tetrabenzofuran, Dihydrophthalate, 9-Octadecaenoic acid and Eicosane were the compounds detected in the extracts. Our findings reveal that Nigrospora spp an endophytic fungi possesses unique chemo diversity of bioactive secondary metabolites that could be utilized for development of new drugs.

Therapeutic Potential of Peucedanum japonicum Thunb. and Its Active Components in a Delayed Corneal Wound Healing Model Following Blue Light Irradiation-Induced Oxidative Stress.

Blue light is reported to be harmful to eyes by inducing reactive oxygen species (ROS). Herein, the roles of Peucedanum japonicum Thunb. leaf extract (PJE) in corneal wound healing under blue light irradiation are investigated. Blue-light-irradiated human corneal epithelial cells (HCECs) show increased intracellular ROS levels and delayed wound healing without a change in survival, and these effects are reversed by PJE treatment. In acute toxicity tests, a single oral administration of PJE (5000 mg/kg) does not induce any signs of clinical toxicity or body weight changes for 15 days post-administration. Rats with OD (oculus dexter, right eye) corneal wounds are divided into seven treatment groups: NL (nonwounded OS (oculus sinister, left eye)), NR (wounded OD), BL (wounded OD + blue light (BL)), and PJE (BL + 25, 50, 100, 200 mg/kg). Blue-light-induced delayed wound healing is dose-dependently recovered by orally administering PJE once daily starting 5 days before wound generation. The reduced tear volume in both eyes in the BL group is also restored by PJE. Forty-eight hours after wound generation, the numbers of inflammatory and apoptotic cells and the expression levels of interleukin-6 (IL-6) largely increase in the BL group, but these values return to almost normal after PJE treatment. The key components of PJE, identified by high-performance liquid chromatography (HPLC) fractionation, are CA, neochlorogenic acid (NCA), and cryptochlorogenic acid (CCA). Each CA isomer effectively reverses the delayed wound healing and excessive ROS production, and their mixture synergistically enhances these effects. The expression of messenger RNAs (mRNAs) related to ROS, such as SOD1, CAT, GPX1, GSTM1, GSTP1, HO-1, and TRXR1, is significantly upregulated by PJE, its components, and the component mixture. Therefore, PJE protects against blue-light-induced delayed corneal wound healing via its antioxidative, anti-inflammatory, and antiapoptotic effects mechanistically related to ROS production.

Identification of a Bitter Peptide Contributing to the Off-Flavor Attributes of Pea Protein Isolates

The aversive bitter taste of pea protein ingredients limits product acceptability. Compounds contributing to the bitter perception of pea protein isolates were investigated. Off-line multi-dimensional sensory-guided preparative liquid chromatography fractionation of a 10% aqueous PPI solution revealed one main bitter compound that was identified by Fourier transform ion cyclotron resonance mass spectrometry and de novo tandem mass spectrometry (MS/MS) sequencing as the 37 amino acid peptide PA1b from pea albumin and further confirmed by synthesis. Quantitative MS/MS analysis reported that the concentration of the bitter peptide was 129.3 mg/L, which was above the determined bitter sensory threshold value of 3.8 mg/L and in agreement with the perceived bitter taste of the sample.

Combining offline high performance liquid chromatography fractionation of peptides and intact proteins to enhance proteome coverage in bottom-up proteomics

Offline peptide separation (PS) using high-performance liquid chromatography (HPLC) is currently used to enhance liquid chromatography–tandem mass spectrometry (LC–MS/MS) detection of proteins. In search of more effective methods for enhancing MS proteome coverage, we developed a robust method for intact protein separation (IPS), an alternative first-dimension separation technique, and explored additional benefits that it offers. Comparing IPS to the traditional PS method, we found that both enhance detection of unique protein IDs to a similar magnitude, though in diverse ways. IPS was especially effective in serum, which has a small number of extremely high abundance proteins. PS was more effective in tissues with fewer dominating high-abundance proteins and was more effective in enhancing detection of post-translational modifications (PTMs). Combining the IPS and PS methods together (IPS+PS) was especially beneficial, enhancing proteome detection more than either method could independently. The comparison of IPS+PS versus six PS fractionation pools increased total number of proteins IDs by nearly double, while also significantly increasing number of unique peptides detected per protein, percent peptide sequence coverage of each protein, and detection of PTMs. This IPS+PS combined method requires fewer LC-MS/MS runs than current PS methods would need to obtain similar improvements in proteome detection, and it is robust, time- and cost-effective, and generally applicable to various tissue and sample types.

Modeling a pesticide remediation strategy for preparative liquid chromatography using high-performance liquid chromatography

BackgroundCannabis sativa L. also known as industrial hemp, is primarily cultivated as source material for cannabinoids cannabidiol (CBD) and ∆9-tetrahydrocannabinol (∆9-THC). Pesticide contamination during plant growth is a common issue in the cannabis industry which can render plant biomass and products made from contaminated material unusable. Remediation strategies to ensure safety compliance are vital to the industry, and special consideration should be given to methods that are non-destructive to concomitant cannabinoids. Preparative liquid chromatography (PLC) is an attractive strategy for remediating pesticide contaminants while also facilitating targeted isolation cannabinoids in cannabis biomass.MethodsThe present study evaluated the benchtop-scale suitability of pesticide remediation by liquid chromatographic eluent fractionation, by comparing retention times of 11 pesticides relative to 26 cannabinoids. The ten pesticides evaluated for retention times are clothianidin, imidacloprid, piperonyl butoxide, pyrethrins (I/II mixture), diuron, permethrin, boscalid, carbaryl, spinosyn A, and myclobutanil. Analytes were separated prior to quantification on an Agilent Infinity II 1260 high performance liquid chromatography with diode array detection (HPLC-DAD). The detection wavelengths used were 208, 220, 230, and 240 nm. Primary studies were performed using an Agilent InfinityLab Poroshell 120 EC-C18 3.0 × 50 mm column with 2.7 μm particle diameter, using a binary gradient. Preliminary studies on Phenomenex Luna 10 μm C18 PREP stationary phase were performed using a 150 × 4.6 mm column.ResultsThe retention times of standards and cannabis matrices were evaluated. The matrices used were raw cannabis flower, ethanol crude extract, CO2 crude extract, distillate, distillation mother liquors, and distillation bottoms. The pesticides clothianidin, imidacloprid, carbaryl, diuron, spinosyn A, and myclobutanil eluted in the first 3.6 min, and all cannabinoids (except for 7-OH-CBD) eluted in the final 12.6 min of the 19-minute gradient for all matrices evaluated. The elution times of 7-OH-CBD and boscalid were 3.44 and 3.55 min, respectively.Discussion7-OH-CBD is a metabolite of CBD and was not observed in the cannabis matrices evaluated. Thus, the present method is suitable for separating 7/11 pesticides and 25/26 cannabinoids tested in the six cannabis matrices tested. 7-OH-CBD, pyrethrins I and II (RTA: 6.8 min, RTB: 10.5 min), permethrin (RTA: 11.9 min, RTB: 12.2 min), and piperonyl butoxide (RTA: 8.3 min, RTB: 11.7 min), will require additional fractionation or purification steps.ConclusionsThe benchtop method was demonstrated have congruent elution profiles using preparative-scale stationary phase. The resolution of pesticides from cannabinoids in this method indicates that eluent fractionation is a highly attractive industrial solution for pesticide remediation of contaminated cannabis materials and targeted isolation of cannabinoids.

Euphorbia Graminea Jacq. (Euphorbiaceae): A Comparative Antimicrobial Evaluations of Stem and Root Extracts

Antimicrobial resistance is a global issue. Euphorbia plants are used locally to treat microbial infections. This study examined the antimicrobial potential of Euphorbia graminea stem and root extracts. The stems and roots extracts of E .graminea were extracted using 80% methanol and tested for antimicrobial activity at concentrations between 4.69-300mg/mL against non-clinical isolates (S. aureus, E. coli, P. aeruginosa, C. albican, A. niger). The active roots extract was fractionated using vacuum liquid chromatographic fractionations (VLC) and the resulting fractions bulked and tested against the organisms at 6.25-100mg/mL. The MIC of extracts and vlc bulked fractions were tested at 0.39-6.25 mg/mL. The root extract recorded higher antimicrobial activities over the stem extract especially against S. aureus and E.coli, hence was fractionated. Among the vlc sub-fractions of the roots extract, fractions A (2) recorded no activity against the test organisms while fractions C (9-10) recorded 7.50 and 3.50 mm against S. aureus and E. coli only at the maximum concentration of 100mg/mL. However, fractions B (3-8) conspicuously gave zones of inhibitions far higher than the other fractions. This study has shown that the roots extract of E. graminea has higher antimicrobial activities more than the stems, further justifying the ethno-botanical potentials of the plant in treating skin infections.