Candida Biofilm Disrupting Ability of Di-rhamnolipid (RL-2) Produced from Pseudomonas aeruginosa DSVP20

Biosurfactant produced from Pseudomonas aeruginosa DSVP20 was evaluated for its potential to disrupt Candida albicans biofilm formed on polystyrene (PS) surfaces in this investigation. P. aeruginosa DSVP20 exhibited optimum production of biosurfactant (5.8 g?L^?1) after 96 h of growth with an ability to reduce surface tension of the aqueous solution from 72 to 28 mN?m^?1. Analysis of purified biosurfactant with FT-IR, ¹H and ¹³C NMR and MALDI-TOF MS revealed it to be di-rhamnolipid (RL-2) in nature. Biofilm disrupting ability of RL-2 (0.16 mg?mL^?1) on Candida cells when checked using XTT reduction assay revealed that about 50 % of the cells remain adhered to 96-well plate after 2 h of treatment, while up to 90 % reduction in pre-formed C. albicans biofilm on PS surface was observed with RL-2 (5.0 mg?mL^?1) in a dose-dependent manner. Microscopic analyses (SEM and CLSM) further confirm the influence of RL-2 on disruption of Candida biofilm extracellular matrix on PS surface which can be exploited as a potential alternative to the available conventional therapies.     

Chemical Composition,Antifungal and Anti-Biofilm Activities of Volatile Fractions of Convolvulus althaeoides L. Roots from Tunisia

The antifungal drugs currently available and mostly used for the treatment of candidiasis exhibit the phenomena of toxicity and increasing resistance. In this context, plant materials might represent promising sources of antifungal agents. The aim of this study is to evaluate for the first time the chemical content of the volatile fractions (VFs) along with the antifungal and anti-biofilm of Convolvulus althaeoides L. roots. The chemical composition was determined by gas chromatography coupled to a flame ionization detector and mass spectrometry. In total, 73 and 86 chemical compounds were detected in the n-hexane (VF1) and chloroform (VF2) fractions, respectively. Analysis revealed the presence of four main compounds: n-hexadecenoic acid (29.77%), 4-vinyl guaiacol (12.2%), bis(2-ethylhexyl)-adipate (9.69%) and eicosane (3.98%) in the VF extracted by hexane (VF1). n-hexadecenoic acid (34.04%), benzyl alcohol (7.86%) and linoleic acid (7.30%) were the main compounds found in the VF extracted with chloroform (VF2). The antifungal minimum inhibitory concentrations (MICs) of the obtained fractions against Candida albicans, Candida glabrata and Candida tropicalis were determined by the micro-dilution technique and values against Candida spp. ranged from 0.87 to 3.5 mg/mL. The biofilm inhibitory concentrations (IBF) and sustained inhibition (BSI) assays on C. albicans, C. glabrata and C. tropicalis were also investigated. The VFs inhibited biofilm formation up to 0.87 mg/mL for C. albicans, up to 1.75 mg/mL against C. glabrata and up to 0.87 mg/mL against C. tropicalis. The obtained results highlighted the synergistic mechanism of the detected molecules in the prevention of candidosic biofilm formation.     

Solid phase extraction of trace amounts of Pb(II) in opium, heroin, lipstick, plants and water samples using modified magnetite nanoparticles prior to its atomic absorption determination

A new, simple, fast and reliable solid-phase extraction method has been developed for separation/preconcentration of trace amounts of Pb(II) using dithizone/sodium dodecyl sulfate-immobilized on alumina-coated magnetite nanoparticles, and its determination by flame atomic absorption spectrometry (FAAS) and graphite furnace atomic absorption spectrometry (GFAAS) after eluting with 4.0?mol?L^?1 HNO₃. Optimal experimental conditions including pH, sample volume, eluent concentration and volume, and co-existing ions have been studied and established. Under the optimal experimental conditions, the preconcentration factor, detection limit, linear range and relative standard deviation of Pb(II) using FAAS technique were 280 (for 560?mL of sample solution), 0.28?ng?mL^?1, 1.4?C70?ng?mL^?1 and 4.6% (for 10?ng?mL^?1, n?=?10), respectively. These analytical parameters using GFAAS technique were 300 (for 600?mL of sample solution), 0.002?ng?mL^?1, 0.006?C13.2?ng?mL^?1 and 3.1% (for 5?ng?mL^?1, n?=?10), respectively. The presented procedure was successfully applied for determination of Pb(II) content in opium, heroin, lipstick, plants and water samples.     

Simultaneous Determination of Buprenorphine, Norbuprenorphine and Naloxone in Human Plasma by LC-MS-MS

A novel sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method simultaneously determined buprenorphine (BUP) and its active metabolite, norbuprenorphine (NBUP), and a coformulant, naloxone was developed, validated and applied successfully in humans. Buprenorphine-d ₄ and norbuprenorphine-d ₃ were used as the internal standard. The analysis was performed on a silica column, and the mobile phase was isocratic and composed of acetonitrile:2 mM ammonium formate in H2O (82:18, v/v). Mass spectrometry employed multiple reaction monitoring modes with transitions of m/z 468.1?C55.2 for BUP, 414.2?C101.2 for NBUP, 328.3?C310.3 for naloxone, 472.1?C59.2 for buprenorphine-d ₄ and 417.2?C101.2 for norbuprenorphine-d ₃. Lower limit of quantification (LLOQ) of the analytical method was 0.05 ng mL^?1 for BUP, 0.1 ng mL^?1 for NBUP and 0.025 ng mL^?1 for naloxone, respectively. The standard calibration curves of BUP, NBUP and naloxone were linear over the concentration range of 0.05?C20 ng mL^?1, 0.1?C20 ng mL^?1 and 0.025?C20 ng mL^?1, respectively. The precisions (RSD) and accuracies (RE) of LLOQ and other QC samples were in acceptable range, with RSD < 20% and RE ± 20% for LLOQ and RSD < 15% and RE within ±15% for QC samples. The method was accurate, precise and specific, and was applied to the pharmacokinetic study of buprenorphine in healthy volunteers.     

Antifungal activity,DNA and lysozyme binding affinity of Pd(II) and Pt(II) complexes bearing N,N-pyridylbenzimidazole ligand

Abstract

The lysozyme- and DNA-binding affinities of the biologically active cisplatin analogues, Pd(II) (1) and Pt(II) (2) complexes of functionalized N,N-pyridylbenzimidazole ligand, are reported. The electronic transitions were studied by time-dependent density functional theory. Complex 1 exhibits interesting antifungal activity against Candida albicans (MIC?=?32?μg?mL^?1; 64?nM) and Cryptococcus neoformans (MIC?=?16?μg?mL^?1, 32?nM). Complex 1 is covalently bound to DNA and lysozyme via the elimination of the chloride ligand(s). When complex 2 interacts with lysozyme, two adduct peaks are observed in the mass spectrum corresponding to binding of Pt(II) ion to lysozyme.     

Simultaneous Determination of Hydrocodone, and Its Two Metabolites in Human Plasma by HPLC�CMS�CMS

A rapid, sensitive and specific assay method has been developed to simultaneously determine human plasma concentrations of hydrocodone and its metabolites, norhydrocodone, hydromorphone, using high-performance liquid chromatography with an electrospray ionization (ESI) tandem mass spectrometry (HPLC?CMS?CMS). Hydrocodone, its metabolites, and internal standard, hydrocodone-d ₃, norhydrocodone-d ₃, hydromorphone-d ₃, were separated from human plasma using solid-phase extraction (Empore MPC-SD Solid Phase Extraction Disk). The eluate was dried, reconstituted and injected into the LC?CMS?CMS system. Chromatographic separation was performed on a Kromasil 100-5SIL-Dimensions C₁₈ column (100 × 2.1 mm, 5.0 ??m, Thermo Hypersil-Keystone, USA) using a gradient mobile phase with 20 mmol L^?1 ammonium formate in water with 0.2% formic acid and 0.1% formic acid in acetonitrile. Detection and quantitation were performed by MS/MS using electrospray ionization and multiple reactions monitoring in the positive ion mode. The calibration curves were linear over the concentration ranges 0.05?C50 ng mL^?1 for hydrocodone (r ² = 0.9991) and norhydrocodone (r ² = 0.9990), and 0.01?C10 ng mL^?1 for hydromorphone (r ² = 0.9990). The limit of quantification was 0.05 ng mL^?1 for hydrocodone and norhydrocodone, and 0.01 ng mL^?1 for hydromorphone. The extraction recovery was above 64.36, 68.51 and 71.78% for hydrocodone, norhydrocodone and hydromorphone. The accuracy was higher than 99.06, 97.70 and 100.07% for hydrocodone, norhydrocodone and hydromorphone. The intra- and inter-day precisions were <5.80, 5.90 and 3.02% for hydrocodone, norhydrocodone and hydromorphone. The method was accurate, sensitive and simple and was successfully applied to a pharmacokinetic study after a single oral administration of hydrocodone bitartrate at a dose of 5 mg in 12 healthy Chinese volunteers.     

Synthesis and biological study of a new series of bifunctional organoiron thio‐ and seleno‐terephthalate derivatives (C5H5)Fe(CO)2ECO(C6H4)COX (E = S,X = R2N,RNH, NH2, OH,Cl; E = Se,X = RNH,RS, RCOO,NH2, OH,Cl)

A new series of bifunctional organoiron thio‐ and seleno‐terephthalate complexes — (η‐C₅H₅)Fe(CO)₂ECO(C₆H₄)COX [E = S; X = C₆H₁₁NH, (C₂H₅)₂N; and E = Se; X = P? CH₃? C₆H₄? NH, C₆H₅? C₂N₂O? S, m? NO₂? C₆H₄? CH?CH? COO] — has been synthesized via the organic transformation reactions of the terephthaloyl chloride precursors η‐(C₅H₅)Fe(CO)₂ECO(C₆H₄)COCl with the desired nucleophiles. These new complexes were characterized by elemental analysis, IR and ¹H NMR spectra. The above complexes, in addition to some other selected analogues, were tested for their antifungal, antibacterial and mutagenic activity. Our results show that all the selenium‐containing compounds have antifungal activity on Candida albicans and antibacterial effects against Bacillus subtilis and Staphylococcus aureus. Four of the six selenium‐containing derivatives exhibited growth inhibitory effects against Pseudomonas aeruginosa and/or Escherichia coli. Sulfur‐containing derivatives elicited activity against C. albicans, and each one of them showed activity against at least one of the bacterial strains that have been used in this investigation. Two selenium‐ and two sulfur‐containing derivatives showed mutagenic activity against one or more than one strain of the Salmonella typhimurium using the Ames test. Copyright © 2001 John Wiley & Sons, Ltd.     

Effectiveness of the Photoactive Dye Methylene Blue versus Caspofungin on the Candida parapsilosis Biofilm in vitro and ex vivo

This research studied the effectiveness of the photoactive compound methylene blue (MB) activated with red LED light (576–672 nm) compared to that of caspofungin (CAS) on 1 Candida albicans and 3 Candida parapsilosis strains. Results were evaluated in terms of SMIC₅₀ for CAS or in PDI (photodynamic inactivation)‐SMIC₅₀ for MB (minimal inhibitory concentration inhibiting sessile biofilm to 50% in comparison to the control without CAS or after irradiation in comparison to the control without MB). While all strains were susceptible to CAS in planktonic form, the SMIC₅₀ was determined to be >16 μg mL^?1 when CAS was added to a 24 h biofilm. However, PDI‐MIC_50s (1.67 mW cm^?2, fluence 15 J cm^?2) were 0.0075–0.03 mmol L^?1. For biofilm, PDI‐SMIC_50s were in the range from 0.7 to 1.35 mmol L^?1. MB concentration of 1 mmol L^?1 prevented a biofilm being formed ex vivo on mouse tongues after irradiation regardless of the application time, in contrast to CAS, which was only effective at a concentration of 16 μg mL^?1 when it was added at the beginning of biofilm formation. PDI seems to be a promising method for the prevention of microbial biofilms that do not respond significantly to conventional drugs.     

In vitro antifungal activity of extracts obtained from Hypericum perforatum adventitious roots cultured in a mist bioreactor against planktonic cells and biofilm of Malassezia furfur

Xanthone-rich extracts from Hypericum perforatum root cultures grown in a Mist Bioreactor as antifungal agents against Malassezia furfur.

Extracts of Hypericum perforatum roots grown in a bioreactor showed activity against planktonic cells and biofilm of Malassezia furfur. Dried biomass, obtained from roots grown under controlled conditions in a ROOTec mist bioreactor, has been extracted with solvents of increasing polarity (i.e. chloroform, ethyl acetate and methanol). The methanolic fraction was the richest in xanthones (2.86 ± 0.43 mg g^? 1 DW) as revealed by HPLC. The minimal inhibitory concentration of the methanol extract against M. furfur planktonic cells was 16 μg mL^? 1. The inhibition percentage of biofilm formation, at a concentration of 16 μg mL^? 1, ranged from 14% to 39%. The results show that H. perforatum root extracts could be used as new antifungal agents in the treatment of Malassezia infections.     

Synthesis and Antifungal Activity of Chimonanthus praecox Derivatives

To search for efficient agricultural antifungal lead compounds, 39 Chimonanthus praecox derivatives were designed, synthesized, and evaluated for their antifungal activities. The structures of target compounds were fully characterized by ¹H NMR, ¹³C NMR, and MS spectra. The preliminary bioassays revealed that some compounds exhibited excellent antifungal activities in vitro. For example, the minimum inhibitory concentration (MIC) of compound b15 against Phytophthora infestans was 1.95 µg mL⁻¹, and the minimum inhibitory concentration (MIC) of compound b17 against Sclerotinia sclerotiorum was 1.95 µg mL⁻¹. Therefore, compounds b15 and b17 were identified as the most promising candidates for further study.     

Flavonoid dihydromyricetin-mediated silver nanoparticles as potential nanomedicine for biomedical treatment of infections caused by opportunistic fungal pathogens

Dihydromyricetin-mediated silver nanoparticles (DMY-AgNPs) were synthesized and their efficacy against fungal pathogens tested in vitro. The shape of DMY-AgNPs appeared to be spherical with size of ~34 nm. Fourier-transform infrared (FT-IR) analysis indicated that –OH and C=O groups were involved in nanoparticle formation. The XRD pattern of DMY-AgNPs showed strong peaks at 38°, 44°, and 64°, corresponding to reflection from (111), (200), and (220) planes. Five opportunistic fungal pathogens, namely Aspergillus fumigatus, Aspergillus niger, Paecilomyces formosus, Candida albicans, and Candida parapsilosis, were isolated from patients suffering from respiratory tract infections. Growth of each fungal strain was inhibited by DMY-AgNPs. The zone of inhibition of DMY-AgNPs against A. fumigatus, A. niger, P. formosus, C. albicans, and C. parapsilosis was 17.6, 19.2, 22.2, 15.8, and 18.5 mm. The minimal inhibitory concentration was found to be 0.83, 0.73, 0.67, 0.95, and 0.89 µg mL^?1, respectively. This is the first report on DMY-AgNPs as an effective antifungal agent. DMY-AgNPs are a potential alternative to commercially available antifungal fungicidals.     

Guanidinylated Amphiphilic Polycarbonates with Enhanced Antimicrobial Activity by Extending the Length of the Spacer Arm and Micelle Self‐Assembly

Nine guanidinylated amphiphilic polycarbonates are rationally designed and synthesized. Each polymer has the same biodegradable backbone but different side groups. The influence of the hydrophobic/hydrophilic effect on antimicrobial activities and cytotoxicity is systematically investigated. The results verify that tuning the length of the spacer arm between the cationic guanidine group and the polycarbonate backbone is an efficient design strategy to alter the hydrophobic/hydrophilic balance without changing the cationic charge density. A spacer arm of six methylene units (CH₂)₆ shows the best antimicrobial activity (minimum inhibitory concentration, MIC = 40 µg mL^?1 against Escherichia coli, MIC = 20 µg mL^?1 against Staphylococcus aureus, MIC = 40 µg mL^?1 against Candida albicans) with low hemolytic activity (HC₅₀ > 2560 µg mL^?1). Furthermore, the guanidinylated polycarbonates exhibit the ability to self‐assemble and present micelle‐like nanostructure due to their intrinsic amphiphilic macromolecular structure. Transmission electron microscopy and dynamic light scattering measurements confirm polymer micelle formation in aqueous solution with sizes ranging from 82 to 288 nm.     

Antioxidant Activities of Polyphenols Extracted from Olive (Olea europaea) of Chamlal Variety

The antioxidant properties of phenolic compounds from olive pulp (PCO) of chamlal variety and those of individual phenolic compounds were evaluated and compared with that of vitamin C (Vit C). The antioxidant activity was measured by the tests of iron reduction and scavenging hydrogen peroxide (H₂O₂). Results showed that all the substances tested exhibit a reducing power. The PCO present activities of iron reduction and H₂O₂ scavenging higher than those of Vit C. The protective effect of PCO against oxidation of lipids and proteins from erythrocyte membranes was studied. The measurement of malondialdehyde generated under oxidative stress conditions induced by hydroxyl radicals generating system revealed that PCO have the most significant protective activity against lipid peroxidation (IC₅₀?=?49.27?±?1.91 μg mL^?1). Paradoxically, Vit C revealed a pro-oxidant effect. Proteins oxidation was evaluated using the H₂O₂/FeSO₄ system and electrophoresis. In the presence of PCO at 1 mg mL^?1, proteins of erythrocyte membranes were protected contrary to those treated with Vit C at the same concentration.     

Synthesis,characterization and antimicrobial activity of a Zn(II) complex with 1-(1H-benzoimidazol-2-yl)-ethanone thiosemicarbazone

A new Zn(II) complex with 1-(1H-benzoimidazol-2-yl)-ethanone thiosemicarbazone [Zn(NO₃)(H₂O)(C₁₀H₁₁N₅S)]NO₃ was prepared and characterized by elemental analyses, FT-IR, ¹H NMR spectroscopy, thermogravimetric analysis (TGA), X-ray diffraction (XRD), and single-crystal X-ray diffraction analysis. The coordination geometry of the pentacoordinated zinc is a distorted square pyramid. The antimicrobial activity of the complex was evaluated using a broth micro-dilution method against a panel of human pathogenic Gram positive, Gram negative bacteria and the yeast Candida albicans. The best inhibitory effect was observed against Enterobacter aerogenes (MIC = 0.031 mg mL^?1).     

Optimal glucose and inoculum concentrations for production of bioactive molecules by Paenibacillus polymyxa RNC-D

The production of antimicrobial metabolites by Paenibacillus polymyxa RNC-D was assessed. Two process variables, glucose and inoculum concentrations, were evaluated at different levels (5?C40 g L^?1, and at ?? _r = 2.5?C5.0 %, respectively), and their effects on biomass formation, minimal inhibitory concentration (MIC) against Escherichia coli, and surface tension reduction (STR) were studied. When the fermentation process was carried out under non-optimised conditions, the biomass, MIC, and STR achieved the following values: 0.6 g L^?1, 1 g L^?1, and 18.4 mN m^?1, respectively. The optimum glucose (16 g L^?1) and inoculum volume ratio (?? _r = 5.0 %) were defined in order to maximise the biomass formation, with a low value of MIC and high STR of extract. The experiments carried out under optimal conditions showed the following values for the dependent variables: biomass concentration 2.05 g L^?1, MIC 31.2 ??g mL^?1, and STR 10.7 mN m^?1, which represented improvement of 241.7 %, 96.9 %, and 41.9 % for the responses of biomass, MIC, and STR, respectively. This is the first recorded study on the optimisation of culture conditions for the production of antimicrobial metabolites of P. polymyxa RNC-D, and constitutes an important step in the development of strategies to modulate the production of antimicrobial molecules by this microorganism at elevated levels.     

Ocular Pharmacokinetic Study of l-Carnosine and N-Acetyl-l-carnosine in Rabbit by Microdialysis Coupled with UPLC-MS-MS

In order to provide useful information for rational drug design, the ocular pharmacokinetics of l-carnosine (CAR) and its acetylized prodrug N-acetyl-l-carnosine (NAC) were investigated. The in vivo microdialysis sampling coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) was developed for continuously simultaneous monitoring of CAR and NAC in rabbit aqueous humor. The measured in vitro recoveries of the probe were 61.3% for CAR and 65.8% for NAC, while in vivo recoveries decreased to 43.1% for CAR and 43.0% for NAC, respectively. The method was sensitive with LLOQ 20.5 ng mL^?1 for CAR and 20.4 ng mL^?1 for NAC. The initial data indicated that the value of C _max and AUC_(0?C??) of NAC were higher than these of CAR (C _max 2305 vs. 1,802 ng mL^?1), (AUC_(0?C??) 1,337 vs. 1,891 ng h mL^?1), which indicated that the NAC exhibited better ocular bioavailability and duration. The method was rapid, specific and sensitive for continuously monitoring of aqueous humor and it was successfully applied to pharmacokinetic studies of CAR and NAC.     

First report of anti-inflammatory chromenyl derivatives from the spineless cuttlefish Sepiella inermis

Abstract

The spineless cuttlefish Sepiella inermis encompasses a major share in the marine fisheries sector, and represents as a culinary delicacy in many cultures. Bioactivity-guided fractionation of methanol:ethyl acetate (MeOH:EtOAc, 1:1) extract of the edible parts of the species ensued in identification of two hexahydro chromenyl analogues namely, methyl 7-ethyl-hexahydro-8a-methyl-2H-chromene-4-carboxylate (1) and methyl 1-acetoxy-hexahydro-3-methyl-3-propyl-1H-isochromene-4-carboxylate (2). The isolated metabolites were checked for their radical scavenging and anti-inflammatory potentials by selective in vitro models. The isochromenyl derivative exhibited potential 2,2-diphenyl-1-picryl-hydrazil and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (IC₅₀?<?0.45?mg mL^?1) radical-scavenging capacities along with pro-inflammatory cyclooxygenase-2 (COX-2) (IC₅₀ 0.75?mg mL^?1) and 5-lipoxygenase (5-LOX) (IC₅₀ 0.77?mg mL^?1) inhibitory activities. The titled compounds displayed the selectivity indices (IC₅₀ anti-COX-1/IC₅₀ anti-COX-2) greater than 1.25, in comparison with synthetic anti-inflammatory drug ibuprofen (0.44), which attributed to their greater selectivity towards inducible pro-inflammatory enzyme COX-2.     

Chlovalicin B,a Chlorinated Sesquiterpene Isolated from the Marine Mushroom Digitatispora marina

As part of our search for bioactive metabolites from understudied marine microorganisms, the new chlorinated metabolite chlovalicin B (1) was isolated from liquid cultures of the marine basidiomycete Digitatispora marina, which was collected and isolated from driftwood found at Vannøya, Norway. The structure of the novel compound was elucidated by spectroscopic methods including 1D and 2D NMR and analysis of HRMS data, revealing that 1 shares its molecular scaffold with a previously isolated compound, chlovalicin. This represents the first compound isolated from the Digitatispora genus, and the first reported fumagillin/ovalicin-like compound isolated from Basidiomycota. Compound 1 was evaluated for antibacterial activities against a panel of five bacteria, its ability to inhibit bacterial biofilm formation, for antifungal activity against Candida albicans, and for cytotoxic activities against malignant and non-malignant human cell lines. Compound 1 displayed weak cytotoxic activity against the human melanoma cell line A2058 (~50% survival at 50 µM). No activity was detected against biofilm formation or C. albicans at 50 µM, or against bacterial growth at 100 µM nor against the production of cytokines by the human acute monocytic leukemia cell line THP-1 at 50 µM.     

Effect of seasonal variation on the chemical composition and antioxidant and antifungal activities of Convolvulus althaeoides L. leaf extracts

The composition of polyphenols, chlorophylls and carotenoids of eight extracts of Convolvulus althaeoides L. leaves, harvested in two different seasons, winter and spring, and extracted by hot extraction method using four solvents (dichloromethane, chloroform, ethyl acetate and ethanol) with increasing polarity, were evaluated along with their antioxidant and antifungal activities. Qualitative and quantitative variations were observed in the composition based on two different high performance liquid chromatography systems, liquid chromatography-photodiode array detection coupled to either atmospheric pressure chemical ionization mass spectrometry or to electrospray ionization mass spectrometry, permitting the identification of 22 polyphenols, 11 chlorophyll derivatives and 10 carotenoid compounds. Polyphenolic compounds were predominant in extracts from leaves collected in winter, whereas pigments were predominant in the spring collections. Antioxidant activities of the extracts were determined by DPPH radical scavenging method, revealing a half inhibition concentration (IC₅₀) ranging from 0.1369 ± 0.0272 mg g⁻¹ to 0.432 ± 0.0018 mg g⁻¹, with no correlation to seasonal fluctuation. Concerning antifungal assays, ethyl acetate and ethanol extracts have been shown to be the most active against dermatophytes (T. rubrum, T. menthagrophytes, M. canis), with inhibiting percentages reaching 100% with 50 mg mL⁻¹. Moreover, ethyl acetate and ethanol extracts showed a maximum inhibition potential with minimum inhibitory/fungicidal concentrations ranging from 0.78 to 6.25 mg mL⁻¹ on Candida spp. cultures. The winter collect of these extracts showed an inhibitory effect of 90% on Candida albicans germ tubes formation, at a concentration of 3.1 mg mL⁻¹. In conclusion, seasonality seems to influence the quality and the quantity of natural substances from leaves of C. althaeoides L., which have major importance on the antioxidant and the antifungal effectiveness.     

HPLC-ELSD Determination of Kryptofix 2.2.2 in the Preparations of 2-Deoxy-2-[18F]fluoro-d-glucose and other Radiopharmaceuticals

A new and simple high-performance liquid chromatography with evaporative light scattering detector method for the determination of Kryptofix 2.2.2 (K-222) in the radiopharmaceuticals of 2-deoxy-2-[¹⁸F]fluoro-d-glucose ([¹⁸F]FDG) and 3′-deoxy-3′-[¹⁸F]fluorothymidine ([¹⁸F]FLT) was developed. A C₁₈ column was used and the mobile phase was 10?% (v/v) methanol and 90?% (v/v) water (0.1?% trifluoroacetic acid, v/v) at a flow rate of 0.2?mL?min^?1. The drift tube temperature was 40?°C. The pressure of nebulizing gas (N₂) was 3.0?bar. The gain was 10. Good separation of K-222 from main related substances could be achieved. Excellent linearity (r ²?=?0.9995) was obtained over the range of 5–100?μg?mL^?1. The precision ranged from 0.68 to 5.16?% (RSD) and the accuracy ranged from ?3.05 to 2.62?% (RE). The limit of detection was 2?μg?mL^?1. This method offers simple, rapid and quantitative detection of K-222, thus making it acceptable for routine determination.