High-performance Liquid Chromatography Diode Research Articles

Development and Validation of a High-Performance Liquid Chromatography Diode Array Detector Method to Measure Seven Neonicotinoids in Wheat.

Neonicotinoids (NEOs), used as insecticides against aphids, whiteflies, lepidopterans, and beetles, have numerous detrimental impacts on human health, including chronic illnesses, cancer, infertility, and birth anomalies. Monitoring the residues in food products is necessary to guarantee public health and ecological balance. The present work validated a new method to measure seven neonicotinoid insecticides (acetamiprid ACT, clothianidin CLT, dinotefuran DNT, imidacloprid IMD, nitenpyram NTP, thiacloprid TCP, and thiamethoxan THT) in wheat. The analytical procedure was based on simple and fast wheat sample cleanup using solid-phase extraction (SPE) to remove interferents and enrich the NEOs, alongside the NEOs' separation and quantification by reverse-phase chromatography coupled with a diode array detector (DAD). The validation process was validated using the accuracy profile strategy, a straightforward decision tool based on the measure of the total error (bias plus standard deviation) of the method. Our results proved that, in the future, at least 95% of the results obtained with the proposed method would fall within the ±15% acceptance limits. The test's cost-effectiveness, rapidity, and simplicity suggest its use for determining the levels of acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam in routine analyses of wheat.

Application of continuous magnetic solid phase extraction in the extraction of tetracycline antibiotics from honey samples

This study aimed to develop a continuous magnetic solid phase extraction method in a narrow bore tube for the extraction of tetracycline antibiotics from honey samples before their determination with high performance liquid chromatography–diode array detector. In this work, a magnetic covalent organic framework is used as the sorbent. The extraction method is performed in a narrow bore tube by adding 25 mg of a magnetic sorbent prepared from melamine, barbituric acid, and Fe3O4 into the diluted honey sample solution. By using a magnet, the sorbent is moved up and down through the narrow bore tube for five times to improve extraction efficiency. After adsorption the analytes, they are eluted from the sorbent surface using 2,2–dimethoxypropane under vortexing. Then, by using a magnet, the eluent containing the extracted analytes are taken and evaporated with continuous flow of nitrogen. Finally, the residue containing the enriched analytes is reconstituted in microliter–level of mobile phase and injected into chromatographic system for quantitative analysis. Based on the outcomes, the method demonstrated good linearity (0.37–1000 ng g−1), high enrichment factors (175–203), good extraction recoveries (70–81 %), low LODs (0.11–0.18 ng g−1) and LOQs (0.37–0.60 ng g−1). The proposed method is effectively applied for quantification of tetracyclines in twenty–four honey samples. Considering the results, none of the honey samples was contaminated with the studied antibiotics. Regarding to the above–mentioned analytical advantages of the proposed method, its simplicity and rapidity, and green synthesis of the magnetic sorbent particles, the suggested method can be used as a routine method for the extraction of tetracyclines from honey samples.

Reaction of cyclamate with hypochlorous acid in vitro and formation of chlorcyclohexylamine and cyclohexylamine

In this work, we investigated the reaction of cyclamate with hypochlorous acid (HOCl) in simulated gastric juice. The reaction products were detected by high-performance liquid chromatography diode array detection (HPLC-DAD) and ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). We also explored the changes in product concentration as a function of reaction time, cyclamate and HOCl concentrations. Cyclamate reacted with hypochlorous acid instantly in the simulated gastric fluid. N, N-dichlorcyclohexylamine and cyclohexylamine were both detected when the HOCl concentration was at millimole. Cyclohexylamine can only be found when HOCl concentration was at micromole. N, N-dichlorcyclohexylamine and cyclohexylamine concentrations both increased when cyclamate concentration increased under the millimole level of HOCl. As an important reactive oxygen species, hypochlorous acid (HClO) is produced in various physiological processes. The abnormal rise of the HClO level is associated with many inflammatory diseases. Chronic gastritis associated with Helicobacter pylori is a multistep, progressive, life-long inflammation. So, chronic gastritis infected with H. pylori may cause cyclamate metabolizing into cyclohexylamine in vivo.

Analyzing the presence of ten synthetic colors in selected food and drink samples using ultra high-performance liquid chromatography diode array detection analysis

AbstractColoring agents in foods and drinks have been popular for centuries. This study aims to analyze the presence of ten synthetic colors (namely, (allura red (E129), amaranth (E123), sunset yellow (E110), tetrazine (E102), fast green (E143), ponceau 4R (New Coccine) (E124), erythrosin B (E127), brilliant blue FCF (E133), brilliant black (E151) and carmoisine (E122))) in food and drink samples using ultra-high-performance liquid chromatography diode array detection (UHPLC-DAD). The present analytical method was carried out using Agilent Poroshell 120 HPH-C18 column, 3 × 100 mm, 2.7 µm, and a mobile phase consisting of 10 mM Na2HPO4, pH 7, mixed with methanol as a time-increment gradient solution until the time was 20 min, then decreased with time until the time was 26 min. The pH was set by orthophosphoric acid at 7 and 5 μL injection volume, 0.50 mL flow rate, and the elution systems were monitored at 428 nm for E102, 518 nm for E124, E110, E129, E122, 530 nm for E151, E127, 622 nm for E143, and E133, respectively. The limit of detection and quantification for all colors ranged from 0.017 to 0.025 and 0.057 and 0.082 mg L−1, respectively. The correlation coefficient values ranged between 0.9991 and 1.0. The selectivity of the assay revealed no interference from other components in the analyzed samples. The percent recovery and precision (intra- and inter-day) of the spiked samples were within the acceptable limits of the ICH guidelines. Five analytical parameters were employed, and the results showed a new, novel, and robust method according to ICH guidelines for analyzing these colors. While most of the investigated food and drinks fell within the accepted range, some fell outside. The current sample preparation and analytical methods are comprehensive and universal for extracting and measuring synthetic colors in various food and drink samples.

Simultaneous Quantification Method of Flavonoids in Jeju Native Citrus from Different Harvest Times Using a High-Performance Liquid Chromatography–Diode Array Detector (HPLC–DAD)

In Jeju-native Citrus, flavonoids are the main contributors to the various types of biological activity, such as antioxidant, antitumor, and anti-inflammatory activity. Thus, we developed simultaneous quantification methods for the analysis of ten bioactive flavonoids in Jeju Citrus fruits (Dangyuja, Gamja, Jigak, Sadugam, and Soyuja) harvested at six different time points using a high-performance liquid chromatography–diode array detector (HPLC-DAD). Separation was performed using a flow rate of 0.8 mL/min, a column temperature of 40 °C, a mobile phase buffer of 0.5% acetic acid, and a detection wavelength of 278 nm. The established analytical method showed good linearity (R2 ≥ 0.9997), precision (inter-day < 0.599%, intra-day < 0.055%), and accuracy (recoveries 92.30–108.80%). The HPLC–DAD method was subsequently applied to analyze flavonoids in Citrus samples. Overall, the quantification results indicated that the compositions and content of flavonoids differed for each Citrus species. The harvesting period also influenced the changes in flavonoid content within each Citrus species. The analytical results with chemometrics revealed that higher flavonoid levels in early-harvested Citrus were derived from the improved fruit size and reduced flavonoid synthesis during maturation. This study provides a practical and reliable method for the analysis of ten flavonoids that can be further utilized in the quality assessment of Jeju Citrus.

An Investigation into Crithmum maritimum L. Leaves as a Source of Antioxidant Polyphenols

Crithmum maritimum, commonly known as rock samphire, is a plant species with a long history of use in traditional medicine and cuisine, especially in the Mediterranean region. Despite its potential therapeutic and commercial applications, the number of studies on this plant species are scanty and sparse. The aim of this study was to optimize the solid–liquid extraction of bioactive compounds from C. maritimum leaves through response surface methodology (RSM) and to comprehensively analyze the resulting extracts. Experimental parameters including liquid-to-solid ratio, solvent composition, extraction time, and temperature were investigated. The results of the RSM revealed a notable variation in the values of the extracted bioactive compounds (polyphenols, carotenoids, and chlorophylls), indicating the selection of optimum extraction conditions. Partial least squares analysis showed that solvent composition and extraction temperature had a significant impact on the extraction of the bioactive compounds. Consequently, these conditions required a 145-min extraction at 80 °C, using a solvent consisting of 45% v/v ethanol, and a 40:1 liquid-to-solid ratio. The optimum extract was analyzed using a range of assays to determine their polyphenol content, their content in chlorophylls and carotenoids, and their antioxidant potential via FRAP and DPPH assays. The total polyphenol content was measured at 15.11 mg gallic acid equivalents (GAE)/g. High-performance liquid chromatography–diode array detector (HPLC-DAD) analysis revealed that chlorogenic acid was the most prevalent polyphenol (9.35 mg/g) in C. maritimum leaves. Kaempferol 3-glycoside (1.81 mg/g), naringin (1.24 mg/g), and hesperidin (0.79 mg/g) were some polyphenolic compounds that had also been quantified. Total carotenoids (0.32 μg/g) and total chlorophylls (0.62 μg/g) were also measured. Multivariate correlation analysis and principal component analysis revealed that total carotenoids and chlorophylls had a strong negative correlation with total polyphenol content. The reducing power (FRAP assay) of the optimized extract had a value of 85.52 μmol ascorbic acid equivalents (AAE)/g, whereas the antiradical activity (DPPH assay) was measured at 25.57 μmol AAE/g. Given the high quantity of polyphenols, these results highlight the potential use of C. maritimum as a source of bioactive compounds in the food and pharmaceutical industries.

Expression analysis of transcription factor (CBF/DREB) gene and biochemical response of Cannabis sativa to cadmium toxicity and foliar application of molybdenum

Plants respond to heavy metals stress by the activation of stress responsive genes to establish biochemical defense mechanisms against the stress. We have investigated the molecular and biochemical response of Cannabis sativa to cadmium (Cd) stress in the presence of molybdenum (Mo) foliar treatments. For this purpose, C. sativa plants were allowed to grow in pots containing Cd polluted soil, for two month duration. Aqueous solutions of ammonium molybdate pentahydrate (0.5, 1.0 and 2.0 ppm) were foliarly sprayed on the plants, at two weeks intervals. Various parameters were inter-correlated i.e., Plant biomass, Cd phyto-accumulation, transcript level/expression of CBF/DREB genes, concentrations of free proline, polyphenolics, and chlorophyll contents. Phenolic compounds were analyzed by High Performance Liquid Chromatography Diode Array Detector (HPLC-DAD) and Cd contents were analyzed by Atomic Absorption Spectrophotometer. Biomass and chlorophyll contents decreased while the concentrations of free proline and polyphenolics increased under Cd stress. Dry biomass of the all the plant parts (roots (1.87 ± 0.22 g), stem (3.28 ± 0.39 g) and leaves (3.54 ± 0.87 g) and free proline in roots (69.00 ± 8.28 µg/g) and leaves (40.56 ± 4.87 µg/g) were most significantly increased with foliar spray of 2.00 ppm Mo. Nineteen (19) different polyphenolic compounds were identified and quantified in leaves and most of them were highly increased in concentration under Cd stress. The highest concentration was noted in Caffeoyl tyramine (1111 µg/g) in plants treated with 2.00 ppm Mo. Transcript level of CBF/DREB genes in control plants were less as compared to Mo treated plants, where a high increase in transcript level of these genes were found. Expression of CBF /DREB genes showed significant positive correlations with Cd accumulation (R2= 0.751) and concentrations of free proline (R2 = 0.62) and polyphenolics (R2 = 0.65) in leaves. The results suggested that Mo increased the expression of CBF/DREB genes and subsequently the concentration of phenolic compounds and free proline to combat plant against the toxicity of high concentration of Cd on plants.

Adjuvant Effects on Pyraclostrobin and Boscalid Residues, Systemic Movement, and Dietary Risk in Garlic under Field Conditions

Adjuvants are supplemental substances added to pesticide tank mixtures to enhance their efficacy by altering the dispersing, emulsifying, spreading, sticking, and wetting properties of the spray mixture. The goals of this study were an assessment of the effectiveness of pyraclostrobin and boscalid against garlic rust; the fungicide’s dissipation and translocation within plant tissue in the presence of commercial adjuvants Dash® HC EC; and the validation of an analytical method for concurrently determining boscalid and pyraclostrobin in green garlic and garlic bulbs. Pyraclostrobin and boscalid retention in green garlic and the dietary exposure risks were also evaluated. The adjuvant Dash® HC EC is an emulsifiable concentrate containing fatty acids, methyl esters, and alkoxylated alcohol–phosphate esters. The test plots were sprayed with the maximum recommended dosage of 400.5 g a.i. ha−1 boscalid and 100.5 g a.i. ha−1 pyraclostrobin. The residues of pyraclostrobin and boscalid in garlic bulb and green garlic samples were determined using a mini-Luke multi-residue solvent extraction method paired with a high-performance liquid chromatography diode array detector (HPLC–DAD). This method is based on the extraction of a homogeneous sample with acetone, petroleum ether, and dichloromethane. Boscalid and pyraclostrobin recoveries in green garlic and garlic bulbs ranged from 71% to 107% on average, with a relative standard deviation (RSD) of 6.8% to 18.1%, and limits of quantification were 0.02 mg/kg. In green garlic and garlic bulbs, boscalid residues ranged from 0.14 to 3.51 mg/kg and pyraclostrobin from lower than LOQ to 1.43 mg/kg, during the experiment. The adjuvant increased the fungicidal efficacy of the fungicides and their retention on green garlic. Although boscalid is considered a nonsystemic fungicide, it showed higher translocation from garlic leaves to garlic bulbs than pyraclostrobin. Regarding the dietary risk assessment, the use of the adjuvant in a tank mixture with boscalid and pyraclostrobin is considered safe 14 days after application, which agrees with the recommended PHI.

A Critical Assessment of Extraction Methodologies for the Valorization of Agricultural Wastes: Polyphenolic Profile and Bioactivity

Different extraction techniques were used to exploit fruit processing residues for their use as a source of phenolic compounds. Three different extraction methods, namely microwave-assisted extraction (MAE), subcritical water extraction (SWE), and maceration (M), were assessed to gauge their respective efficacies. Total phenolic content (TPC) and total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and radical scavenging activity ABTS assay were evaluated. High-performance liquid chromatography was used to assess the polyphenolic profile. MAE was the extraction technique that allowed the highest recovery of polyphenolic compounds. Concerning the fruit by-products analyzed, the extract of pomegranate peels obtained using M60C and MAE had the highest TPC (313 ± 24 mg GAE/g dry weight (dw)) and TFC (36.0 ± 2.8 mg EE/g dw), respectively, and the highest antioxidant activity (FRAP = 740 ± 67 mg AAE/g dw and ABTS (628 ± 27 mg TE/g dw) corresponded to M60C. The phenolic composition obtained for this sample using high-performance liquid chromatographic–diode array detection (HPLC–DAD) showed that gallic acid, protocatechuic acid, β-resorcylic acid, (+)-cathechin, and rutin were the main phenolics found. The findings underscore the capacity of agricultural by-products to act as a source of phenolic compounds. This offers a feasible solution to enhance the nutritional content in food while simultaneously minimizing environmental waste.

Fast surface floating organic droplets based dispersive liquid‐liquid microextraction for trace enrichment of multiclass pesticide residues from different fruit juice samples followed by high performance liquid chromatography–diode array detection analysis

AbstractThis study was designed to enable the development of a simple, fast, and environmentally friendly analytical technique utilizing dispersive liquid‐liquid microextraction based on surface floating organic droplets for selective and quantitative enrichment of trace level pesticide contaminants from different fruit juice samples for subsequent detection by high performance liquid chromatography, combined with a diode array detector. The selective extraction was necessitated in order to isolate the seven multiclass pesticide residues frequently occurring in fruit juice samples. The effects of experimental parameters such as pH of sample solution, type and volume of extraction and dispersive solvents, ionic strength and extraction time were optimized. The optimized method was validated using spiked blank sample and satisfactory results for accuracy, with recoveries ranging from 87.23% to 99.45%, with %relative standard deviation between 1.37 and 8.39, precision in terms of %relative standard deviation ≤ 10.78 and linearity at concentration levels from 3 to 1500 ng/ml, which corresponded with correlation coefficients ≥ 0.998. The limits of detection and the limits of quantification were ranged from 1.3×10−2 to 5.3×10−2 and 4.2×10−2 to 1.8×10−1 μg/L, respectively. At the end, the method was successfully applied to analyze real fruit juice samples and target analytes were not detected in real samples.

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.

Magnet Integrated Fabric Phase Sorptive Extraction for the Extraction of Resin Monomers from Human Urine Prior to HPLC Analysis

In this work, a method for the simultaneous determination of four resin monomers: Bisphenol A, bisphenol A methacrylate glycidate, triethyleneglycol-dimethacrylate, and urethane dimethacrylate, from human urine using magnet integrated fabric phase sorptive extraction (MI-FPSE), followed by high performance liquid chromatography (HPLC) diode array detection (HPLC-DAD), is presented. MI-FPSE is a novel configuration of FPSE that incorporates the stirring and extraction mechanism into one device, resulting in an improved extraction kinetic factor. FPSE is a green sample preparation technique that uses a flexible surface, such as cellulose, coated with a polymeric material using sol–gel technology. Poly(tetrahydrofuran) (PTHF) material was selected, due to its higher efficiency in terms of recovery rate among the studied MI-FPSE membranes. Optimization of the extraction process was performed based on several extraction and elution parameters. The method was validated for its linearity, selectivity, accuracy, precision, and stability of the samples. For the four compounds, the LOD and LOQ were 0.170 ng/μL and 0.050 ng/μL, respectively. The relative standard deviation of the method was less than 9.8% and 11.9%, for the within-day and between-day precision, respectively. The relative recoveries were between 85.6 and 105.2% in all cases, showing a good accuracy. The effectiveness of the proposed method was confirmed through successful application to the bioanalysis of real urine samples.

Dispersive Liquid - Liquid Micro extraction Technique Coupled with High Performance Liquid Chromatography Diode Array Detector for the Determination of Sulfonamides in Water Samples

A dispersive liquid-liquid microextraction combined with high-performance liquid chromatography with diode array detection (DLLME-HPLC-DAD) has been developed for the determination of 15 sulfonamides in water samples. The effects of experimental parameters on the performance of the method such as the type and volume of extraction and disperser solvents, pH of the solution, effect of salt, and centrifugation time were evaluated, and optimum conditions were established. Under these optimum conditions, linearity was found in the range of 2 - 1 000 μg L-1 with regression coefficients better than 0.9990. The limit of detection (LOD) and limit of quantification (LOQ) values were in the range of 0.6 - 7.8 μg L-1 and 1.5 - 8.3 µg L-1, respectively. Intra-day and inter-day precision ranged from 1.5 - 9.7% and from 3.1 - 9.9%, respectively. The accuracy of the method were also evaluated in samples from three different sources of water (river water, groundwater, and tap water) were found to range from 70.5 to 103.9% with %RSD values ranging from 0.5 to 9.8%, with the exception of sulfaguanidine in river water and groundwater where the percentage recovery ranged from 54.3 - 84.9% at the two higher concentration levels (100 and 400 µg L-1).Furthermore, the effect of sample matrix on the extraction of target compounds was evaluated using the relative recovery (%RR) of the samples from three different water sources relative to the ultrahigh purity (UHP) water sample spiked with target analytes, and found in the range of 70.3 - 106.1%.The proposed method was successfully applied to wastewater treatment plant samples and Sulfisoxazole (SSO) was detected in the influent samples in the range of 14.8 - 17.8 µg L-1 with corresponding %RSD, (n = 5) value of 1.6% to 1.9%.The results indicated that the proposed method is effective for the extraction and determination of the sulfonamides in water samples.

Stability of Important Veterinary Antibiotics Amoxicillin, Sulfadiazine, and Trimethoprim in Practice-Relevant Model Solutions

Due to the frequent use of veterinary drugs in animal husbandry, it is important to know their environmental behavior. In this context, little attention has been paid to the stability of the active ingredients in solutions prepared for administration. This is particularly problematic for antibiotics that trigger resistance when administered subtherapeutically. In order to investigate a possible influence of the preparation and storage of veterinary drugs on compound stability, three widely used antibiotics (amoxicillin, sulfadiazine, trimethoprim) were prepared in different model solutions. Depending on their individual stabilities, the incubation period lasted up to 70 days. Samples were analyzed at regular intervals by high-performance liquid chromatography-diode array detection and ultraviolet spectrophotometry. Following official recommendations, the investigations covered various parameters, e.g., pH, buffer substances, influence of light, and temperature. Sulfadiazine was incubated together with trimethoprim at concentrations of 120 mg L-1 and 80 mg L-1 for 70 days. Both compounds proved to be very stable under all experimental conditions and between 92 and 100% of the active ingredients remained. In 0.1% formic acid, a transformation product was found with less than 5% of the parent substance. In contrast, amoxicillin (500 mg L-1) was instable in almost all solutions under investigation. Within 17 days, the concentration of AMO decreased to 72% in ultrapure water. With the exception of a physiological saline solution, the amount of amoxicillin dropped below 10% or even below the detection limit. Thus, a physiological saline solution is best suited for the storage of dissolved amoxicillin for later administration.

Preparation of Bioactive De-Chlorophyll Rhein-Rich Senna alata Extract.

Senna alata leaves display various biological activities as a result of their rhein and phenolic composition. The objective of this study was to develop bioactive de-chlorophyll rhein-rich S. alata extracts. The rhein content was quantified using a validated high-performance liquid chromatography-diode array detection (HPLC-DAD) method. The best process parameters for maximizing rhein were established using ultrasound-assisted extraction (UAE). The optimal conditions for the parameters were determined using the Box-Behnken design (BBD); 95% v/v ethanol was used as the extraction solvent at 59.52 °C for 18.4 min with a solvent-to-solid ratio of 25.48:1 (mL/g) to obtain the predicted value of rhein at 10.44 mg/g extract. However, the color of the rhein-rich extract remained dark brown. For the removal of chlorophyll, liquid-liquid extraction with vegetable oils and adsorption with bleaching agents were employed. The bleaching agents were significantly more effective at removing chlorophyll and had less of an effect on the reduction in rhein content than vegetable oils. The presence of rhein and phenolics in the de-chlorophyll extracts might be responsible for their antioxidant, anti-inflammatory, and antibacterial activities. These findings indicate that rhein-rich extract and its de-chlorophyll extracts possess sufficient biological activities for the further development of cosmeceuticals and pharmaceuticals.

Composition characterization and biological activity study of Achillea vermicularis extract and extract-loaded nanoparticles.

Antiproliferative activity of Achillea vermicularis extracts was calculated on glial (C6) and keratinocyte (HaCaT) cell lines using XTT assay. It was observed that all extracts of A. vermicularis at the determined concentration were not cytotoxic in HaCaT cell lines. The nanoparticles (NPs) of the extract with the best cytotoxic activity was prepared, and necessary characterization studies were performed. Results showed that NP containing the extract has a lower IC50 value and more cytotoxic activity in C6 cells compared to the only extract. Furthermore, the antiepileptic potentials of these substances were explored in this study. The effect of A. vermicularis extracts on the enzyme activities of carbonic anhydrase I and II isoenzymes (hCA I and hCA II) was measured using spectrophotometry to achieve this goal. A. vermicularis extracts demonstrated high inhibitory activities compared to standard inhibitor (acetazolamide, AAZ), with IC50 values in the range of 5.04-10.8μg/ml for hCA I, and 5.40-9.22μg/ml for hCA II. High-performance liquid chromatography diode array detector (HPLC-DAD) was used in this investigation to assess the main chemicals found in the extract and NPs. The results showed that the ethanol extract (157.636μg/mg extract) and NPs (4.631μg/mg extract) had a significant amount of the 8-hydroxy salvigenin component.

Anticancer and Antioxidant Activity of Water-Soluble Polysaccharides from Ganoderma aff. australe against Human Osteosarcoma Cells.

Wild mushrooms have gained great importance for being a source of biologically active compounds. In this work, we evaluate the anticancer and antioxidant activity of a water-soluble crude polysaccharide extract isolated from the fruiting bodies of the Ganoderma aff. australe (GACP). This mushroom was collected in San Mateo (Boyacá, Colombia) and identified based on macroscopic and microscopic characterization. GACP was characterized by UV-Vis spectroscopy, Fourier-transform infrared spectroscopy, high-performance liquid chromatography-diode array detector, and nuclear magnetic resonance. The antiradical and antioxidant activity were evaluated by different methods and its anticancer activity was verified in the osteosarcoma MG-63 human cell line. Chemical and spectroscopic analysis indicated that GACP consisted of β-D-Glcp-(1→, →3)-β-D-Glcp-(1→ and α-D-Glcp-(1→ residues. The results of the biological activity showed that GACP exhibited high antioxidant activity in the different methods and models studied. Moreover, the results showed that GACP impaired cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay) and cell proliferation (clonogenic assay) in a dose-response manner on MG-63 cells. The findings of this work promote the use of mushroom-derived compounds as anticancer and antioxidant agents for potential use in the pharmaceutical and food industries.

Nanoporous Ethylenediamine-Functionalized Metal–Organic Framework MIL-101 for the Removal of Proteins and Antipsychotic Drugs from Serum Samples

The development of functional materials for efficient protein removal and antipsychotic drug extraction in biological samples is attracting great interest. In this study, a nanoscale ethylenediamine-functionalized chromium-based metal–organic framework (MOF) (MIL-101-ED) was designed, synthesized, and applied for the one-step removal of proteins and the extraction of antipsychotic drugs in serum samples. The introduction of ethylenediamine in the MIL-101 nanoparticle (NP), in which amino and carbon chains can enhance the hydrogen-bond interaction and π–π interaction between functionalized MOFs and antipsychotic drugs, i.e., risperidone, etc. The antipsychotic drugs were ionized and amino groups were protonated at pH = 3, due to electrostatic interaction, and the negatively charged drugs were adsorbed on the surface or even entered the pores of MOFs. The adsorption efficiencies of the six kinds of drugs ranged from 80.1 to 101.4%. Owing to the exclusion action, proteins were excluded from MIL-101-ED and the exclusion rate ranged from 91.9 to 94.0%. Furthermore, MIL-101-ED can be easily regenerated and reused four times without a significant reduction in the protein exclusion rate and adsorption capacity. Coupled with high-performance liquid chromatography–diode array detection (HPLC–DAD) strategy, the application of MIL-101-ED for extracting the antipsychotic drugs and their metabolites in serum samples was developed. The recoveries of antipsychotic drugs and their metabolites in serum ranged from 83.2 to 110.8%, and the related detection limits were all <5.55 ng mL–1. Compared to the traditional extraction methods, the pretreatment steps of MIL-101-ED have the advantages of easy operation and reducing the environmental pollution. Here, we propose that MIL-101-ED, as a kind of restricted access media nanophase material, can be applied to extract antipsychotic drugs in serum samples.

Qualitative and quantitative phytochemicals of leaves, bark and roots of Antiaris toxicaria Lesch., a promising natural medicinal plant and source of pesticides

Antiaris toxicaria Lesch. of the Moraceae family is a tree that grows endemic to Indonesia and has a height of about 20-30 m. This study aimed to screen the phytochemical constituents of leaf, bark and its root. The plant materials were collected from the Samarinda Botanical Garden. Latex of this species in Indonesia is known as a source of blowpipe poison. In other countries Antiaris sp. plant parts (leaves, bark and seeds) are used in ethnobotany practice as raw material for traditional medicine. The leaves, bark and seeds of this plant are used in the treatment of syphilis, leprosy, cancer and used as laxatives for sore throats. Screening of the phytochemical constituents of the samples began by tracing the macromolecules of alkaloids, steroids, tannins, phenolic compounds, flavonoids and saponins with various tests. Alkaloids, saponins, tannins, phlobatnins, flavonoids and terpenoids were detected in the chemical analysis. High performance liquid chromatography diode array detection (HPLC-DAD) was conducted. HPLC screening of A. toxicaria extracts revealed the presence of galic acid, catechins, chlorogenic acid, caffeic acid, ellagic acid, epigallocatechin, routine, isoquercitrin, quercitrin, quercetin and kaempferol. The study revealed the array of secondary metabolites present in the plant that can be used in medicinal preparations and will be candidate species for developing a natural insecticide.

Combination of mixed mode dispersive solid phase extraction with magnetic ionic liquids based dispersive liquid–liquid microextraction for the extraction of anticoagulant drugs from urine samples

In this report, combination of a mixed mode dispersive solid phase extraction with dispersive liquid–liquid microextraction has been performed in order to the effective isolation of three anticoagulant drugs (anagrelide, betrixaban, and apixaban) from urine sample. The extracted drugs were quantified by high performance liquid chromatography–diode array detector. Firstly, analytes were adsorbed onto a mixture of octadecylsilane, graphitized carbon black, and primary secondary amine sorbents which their composition was obtained by a simplex centroid design. To increase the adsorption efficiency, sonication of the solution and sorbents mixture was performed. The loaded analytes onto the sorbent surface were washed employing a water–miscible organic solvent which was utilized as a disperser in microextraction process. The microextraction step was done using a magnetic ionic liquid (as an extractant) and the droplets were sedimented at the bottom of tube with the aid of an external magnetic field. Satisfactory analytical results such as wide linear ranges (0.21–250 µg/L), high enrichment factors (223–247) and extraction recoveries (76–84 %), and low limits of detection (0.04–0.06 µg/L) and quantification (0.16–0.21 µg/L) were obtained using the optimum experimental situations. Lastly, the offered procedure was utilized for the quantification of the opted analytes in urine samples.