In Vitro Assessment of Organic and Residual Fractions of Nematicidal Culture Filtrates from Thirteen Tropical Trichoderma Strains and Metabolic Profiles of Most-Active

The nematicidal properties of Trichoderma species have potential for developing safer biocontrol agents. In the present study, 13 native Trichoderma strains from T. citrinoviride, T. ghanense (2 strains), T. harzianum (4), T. koningiopsis, T. simmonsii, and T. virens (4) with nematicidal activity were selected and cultured in potato dextrose broth to obtain a culture filtrate (CF) for each. Each CF was partitioned with ethyl acetate to obtain organic (EA) and residual filtrate (RF) fractions, which were then tested on second-stage juveniles (J2s) of the nematodes Meloidogyne javanica and M. incognita in a microdilution assay. The most lethal strains were T. harzianum Th43-14, T. koningiopsis Th41-11, T. ghanense Th02-04, and T. virens Th32-09, which caused 51–100% mortality (%M) of J2s of both nematodes, mainly due to their RF fractions. Liquid chromatography–diode array detector-electrospray-high resolution mass spectrometry analysis of the most-active fractions revealed sesquiterpene and polyketide-like metabolites produced by the four active strains. These native Trichoderma strains have a high potential to develop safer natural products for the biocontrol of Meloidogyne species.

Phytochemical Characterization for Quality Control of Phyllostachys pubescens Leaves Using High-Performance Liquid Chromatography Coupled with Diode Array Detector and Tandem Mass Detector

Phyllostachys pubescens leaves are cultivated in a number of Asian countries and have been used for antipyretic and diuretic effects since ancient times, especially in Korea. The purpose of this study was to develop and validate of analytical method for quality control of P. pubescens leaves using high-performance liquid chromatography with diode array detector (HPLC–DAD) and liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection. HPLC–DAD analysis was conducted with a Gemini C18 column, and distilled water–acetonitrile (both with 0.1% (v/v) formic acid) mobile-phase system. For the LC–MS/MS analysis, all markers were separated with a Waters ACQUITY UPLC BEH C18 column and gradient flow system of distilled water containing 0.1% (v/v) formic acid and 5 mM ammonium formate–acetonitrile. In both method, major components were detected at 2.13–11.63 mg/g (HPLC–DAD) and 0.12–19.20 mg/g (LC–MS/MS). These methods were validated with respect to linearity (coefficient of determination >0.99), recovery (95.22–118.81%), accuracy (90.52–116.96), and precision (<4.0%), and were successfully applied for the quantitative analysis of P. pubescens leaves.

Characterization of Pigments from Malus domestica Leaves for Wool Dyeing

The study will present results of chemical analysis of mass spectrometry, evaluating the content of pigments in M. domestica leaves dyeing solution and in dyed woollen yarn nowadays. Yarn was dyed with fresh leaves of M. domestica tree. Before dyeing the yarn was treated with potassium aluminium sulphate mordant. Dyestuffs from the yarn were extracted in compliance with modified hydrochloric method of hydrolysis. Extracts were analysed by ultra high-performance liquid chromatography coupled with a diode array detector and mass spectrometry (UHPLC-DAD-MS) for identification of the pigments. Phytochemical screening of dyeing solution of the M. domestica leaves showed presence of dihydrochalcones (phloretin, phlorizin), flavan-3-ols (dimeric B type proanthocyanidin, catechin, epicatechin, gallocatehin, epigallocatechin), flavanols (quercetin derivatives: hyperoside, rutin, quercetrin), phenolic acids and other compounds.

Quantification of aniline and N-methylaniline in indigo

Aniline and N-methylaniline are common contaminants in commercially produced indigo. It is known, that commercially produced indigo contains up to 0.6% aniline and 0.4% N-methylaniline by weight and indigo dye shows a small mutagenic effect, most probably due to the presence of these contaminants. The present work describes a new and powerful analytical method to determine the concentration of these contaminants in indigo. This method is based on the transformation of water insoluble indigo into soluble leucoindigo and allows therefore the acidic extraction of the aromatic contaminants. This transformation step is essential, because the main part of these contaminants are strongly included in the indigo crystals. The amount of extracted aniline and N-methylaniline from the leucoindigo solution was quantified with high performance liquid chromatography (HPLC, combined with a photo diode array detector). A possible accumulation of the aromatic amines at the indigo crystal surface was investigated using FTIR and by adsorption studies. Therefore this method allows an accurate monitoring of these toxic by-products in the indigo dye, which is important for an economic and environmental assessment of the denim production.

Development and Validation of RP-HPLC Method for Determation of Acyclovir in Ointment

The analytical method was developed and validated for determination of acyclovir in ointment by High performance liquid chromatography. The separation was carried out on Luna C18 column (250 × 4.6mm × 5µ). The mobile phase consists of water: acetonitrile in the ratio 88:12 at flow rate 0.8ml/min with diode array detector wavelength at 254nm.The column temperature was adjusted at 30ºC±40ºC with injection volume 20µl.The retention time of acyclovir was 4.747min. The linearity of the calibration curve was linear over the concentration range 80-120µg/ml (r2=0.9996). The validation was carried out as per ICH guidelines. The development method was easy, rapid, linear, precise, accurate and consistent.

Clitorienolactones and Isoflavonoids of Clitorea ternatea Roots Alleviate Stress-Like Symptoms in a Reserpine-Induced Zebrafish Model

Clitorea ternatea has been used in Ayurvedic medicine as a brain stimulant to treat mental illnesses and mental functional disorders. In this study, the metabolite profiles of crude C. ternatea root extract (CTRE), ethyl acetate (EA), and 50% aqueous methanol (50% MeOH) fractions were investigated using ultrahigh-performance liquid chromatography–diode array detector–tandem mass spectrometry (UHPLC–DAD–MS/MS), while their effect on the stress-like behavior of zebrafish, pharmacologically induced with reserpine, was investigated. A total of 32 compounds were putatively identified, among which, a series of norneolignans, clitorienolactones, and various flavonoids (flavone, flavonol, isoflavone, and isoflavanone) was found to comprise the major constituents, particularly in the EA and 50% MeOH fractions. The clitorienolactones, presently unique to the species, were present in both the free and glycosylated forms in the roots. Both the EA and 50% MeOH fractions displayed moderate effects on the stress-induced zebrafish model, significantly decreasing freezing duration and elevating the total distance travelled and average velocity, 72 h post-treatment. The results of the present study provide further evidence that the basis for the use of C. ternatea roots in traditional medicine to alleviate brain-related conditions, such as stress and depression, is attributable to the presence of clitorienolactones and the isoflavonoidal constituents.