Preparative separation of flavonoids from the medicinal plant Davilla elliptica St. Hill. by high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) is a major tool for the fast separation of natural products from plants. It was used for the preparative isolation of the flavonoid monoglucosides present in the aerial parts of the Davilla elliptica St. Hill. (Dilleniaceae). This species is used in Brazilian folk medicine for the treatment of gastric disorders. The optimum solvent system used was composed of a mixture of ethyl acetate-n-propanol-water (140:8:80, v/v/v) and led to a successful separation of quercetin-3-O-alpha-L-rhamnopyranoside and myricetin-3-O-alpha-L-rhamnopyranoside in approximately 3.0 hours with purity higher than 95%. Identification was performed by ¹H NMR, 13C NMR and HPLC-UV-DAD analyses.

Preparative separation and purification of sesquiterpenoids from tussilago farfara l. by high-speed counter-current chromatography

Preparative high-speed counter-current chromatography (HSCCC) was successfully applied for separation and purification of sesquiterpenoids from an extract of Tussilago farfara L. with a two-phase solvent system composed of n-hexane-ethyl acetate- methanol-water (1:0.5:1.1:0.3, v/v/v/v). The separation produced a total of 32 mg of tussilagone, 18 mg of 14-acetoxy-7β-(3'-ethyl cis-crotonoyloxy)-lα-(2'-methyl butyryloxy)-notonipetranone and 21 mg of 7β-(3'-ethyl cis-crotonoyloxy)-lα-(2'- methyl butyryloxy)-3,14-dehydro-Z-notonipetranone from 500 mg of the crude extract in one step separation with the purity of 99.5, 99.4 and 99.1%, respectively, as determined by HPLC. The structures of these compounds were identified by ESI-MS, ¹H-NMR and 13C-NMR.

Extraction and isolation of dictamnine, obacunone and fraxinellone from Dictamnus dasycarpus Turcz. by supercritical fluid extraction and high-speed counter-current chromatography

Supercritical fluid extraction was used to extract active compounds from the Chinese traditional medicinal D. dasycarpus under the pressure of 30 MPa and temperature of 45 ºC. Further separation and purification was established by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:0.8:1.3:0.9, volume ratio). The separation yielded a total of 47 mg of dictamnine, 24 mg of obacunone and 83 mg of fraxinellone from 1.0 g of the crude extract in one step separation with the purity of 99.2, 98.4 and 99.0%, respectively, as determined by HPLC. The chemical structures of these compounds were identified by ESI-MS, IR, ¹H-NMR and 13C-NMR.

Preparative separation and purification of bufadienolides from ChanSu by high-speed counter-current chromatography combined with preparative HPLC

Eight bufadienolides were successfully isolated and purified from ChanSu by high-speed counter-current chromatography (HSCCC) combined with preparative HPLC (prep-HPLC). First, a stepwise elution mode of HSCCC with the solvent system composed of petroleum ether - ethyl acetate - methanol - water (4:6:4:6, 4:6:5:5, v/v) was employed and four bufadienolides, two partially purified fractions were obtained from 200 mg of crude extract. The partially purified fractions III and VI were then further separated by prep-HPLC, respectively, and another four bufadienolides were recovered. Their structures were confirmed by ESI-MS and ¹H-NMR spectra.

Preparative separation of polyphenols from artichoke by polyamide column chromatography and high-speed counter-current chromatography

An efficient method for the rapid separation and purification of polyphenols from artichoke by polyamide column chromatography in combination with high-speed counter-current chromatography (HSCCC) was successfully built. The crude ethanol extracts from dry artichoke were first pre-separated by polyamide column chromatography and divided in two parts as sample 1 and sample 2. Then, the samples were further separated by HSCCC and yielded 7.8 mg of chlorogenic acid (compound I), 24.5 mg of luteolin-7-O-β-D-rutinoside (compound II), 18.4 mg of luteolin-7-O-β-D-glucoside (compound III), and 33.4 mg of cynarin (compound IV) with purity levels of 92.0%, 98.2%, 98.5%, and 98.0%, respectively, as determined by high-performance liquid chromatography (HPLC) method. The chemical structures of these compounds were identified by electrospray ionization-mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR).

Separation and purification of three stilbenes from the radix of Polygonum cillinerve (Nakai) Ohwl by macroporous resin column chromatography combined with high-speed counter-current chromatography

An effective method for the rapid separation and purification of three stilbenes from the radix of Polygonum cillinerve (Nakai) Ohwl by macroporous resin column chromatography combined with high-speed counter-current chromatography (HSCCC) was successfully established. In the present study, a two-phase solvent system composed of chloroform-n-butanol-methanol-water (4:1:4:2, v/v/v/v) was used for HSCCC separation. A one-step separation in 4 h from 150 mg of crude extract produced 26.3 mg of trans-resveratrol-3-O-glucoside, 42.0 mg of pieceid-2"-O-gallate, and 17.9 mg of trans-resveratrol with purities of 99.1%, 97.8%, and 99.4%, respectively, as determined by high-performance liquid chromatography (HPLC). The chemical structures of these compounds were identified by nuclear magnetic resonance (NMR) spectroscopy.

Performance of spray nozzles in land applications with high speed

The aim of this study was to evaluate different spray nozzles for land applications in high speed on the coverage and deposit in soybean plants pulverization. It was evaluated the AXI 110 04 plane jet nozzles operated at speed of 4.17m.s-1 (control), the grey APE and the AXI 110 08 plane jets, and the TD HiSpeed 110 06 and AXI TWIN 120 06 twin jets, at speed of 9.72m.s-1. The application volume was fixed in 120L ha-1. The application efficiency was evaluated by two different methods: analysis of the coverage area using fluorescent pigment and UV light and analysis of deposits through the recovery and quantification of FD&C N°1 brilliant blue marker by spectrophotometry. Both analyses were done in samples collected from top, middle and bottom parts of the plants. The spray nozzles showed differences in coverage and deposit pattern, so in the top part, the coverage was increased with smaller drops and the deposits were increased with medium drops. In the other parts of the plants, there were no statistical differences between the treatments for both coverage and deposits. The displacement speed did not influence the application efficiency for nozzles with the same drop pattern, and the obtained spray coverage and deposits at the medium and bottom parts of the plants were less than 50% of that found at the top of the soybean plants.

Curving simulation and stability of a creep-controlled wheelset for high speed rail-vehicles

Higher travel speeds of rail vehicles will be possible by developing sophisticated top performance bogies having creep-controlled wheelsets. In this case the torque transmission between the right and the left wheel is realized by an actively controlled creep coupling. To investigate hunting stability and curving capability the linear equations of motion are written in state space notation. Simulation results are obtained with realistic system parameters from industry and various controller gains. The advantage of the „creep-controlled wheelset" is discussed by comparison the simulation results with the dynamic behaviour of the special cases „solid-axle wheelset" and „loose wheelset" (independent rotation of the wheels). The stability is also investigated with a root-locus analysis.

On the development of an unstructured grid solver for inert and reactive high speed flow simulations

An unstructured grid Euler solver for reactive compressible flow applications is presented. The method is implemented in a cell centered, finite volume context for unstructured triangular grids. Three different schemes for spatial discretization are implemented and analyzed. Time march is implemented in a time-split fashion with independent integrators for the flow and chemistry equations. The capability implemented is tested for inert flows in a hypersonic inlet and for inert and reactive supersonic flows over a 2-D wedge. The results of the different schemes are compared with each other and with independent calculations using a structured grid code. The strengths and the possible weaknesses of the proposed methods are discussed.

A modified contour error controller for a high speed XY table

This article deals with a contour error controller (CEC) applied in a high speed biaxial table. It works simultaneously with the table axes controllers, helping them. In the early stages of the investigation, it was observed that its main problem is imprecision when tracking non-linear contours at high speeds. The objectives of this work are to show that this problem is caused by the lack of exactness of the contour error mathematical model and to propose modifications in it. An additional term is included, resulting in a more accurate value of the contour error, enabling the use of this type of motion controller at higher feedrate. The response results from simulated and experimental tests are compared with those of common PID and non-corrected CEC in order to analyse the effectiveness of this controller over the system. The main conclusions are that the proposed contour error mathematical model is simple, accurate, almost insensible to the feedrate and that a 20:1 reduction of the integral absolute contour error is possible.

Serodiagnosis of Babesia equi in horses submitted to exercise stress

A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.

Methylene blue vital staining for Trypanosoma cruzi trypomastigotes and epimastigotes

The morphological identification of Trypanosoma cruzi is currently considered to have a high specificity, but its sensitivity, which depends on the volume of the sample examined, is rather low. Trypanosome developmental stages suspended in blood, reduviid feces, and culture media are routinely searched for by means of fresh film examination (about 2 µL). High speed centrifugation of blood samples separates the buffy coat, where most trypomastigotes concentrate. As the parasites are transparent and colorless, their detection is mostly dependent on their motility. The fluorescent vital stain acridine orange has been used to enhance image contrast, as exemplified by the QBC (Quantitative Buffy Coat) technique. Staining blood, buffy coat, reduviid feces, and culture media samples with methylene blue (also a vital dye) is a means of producing sharp, well contrasted images of motile or non-motile T. cruzi developmental stages, only standard laboratory microscopes being required. Slides previously coated with a thin layer of methylene blue are used to stain fresh blood films. Photomicrographs exemplify the results of methylene blue staining applied to living and fixed parasites.

Rapid detection by transmission electron microscopy of mycoplasma contamination in sera and cell cultures

Transmission electon microscopy has been employed for the rapid detection of mycoplasma in sera and cell cultures. High speed centrifugation of sera or low speed centrifugation of cell debris, followed by negative staining of the resuspended pellet, detected mycoplasma contamination more frequently than a culture method followed by direct fluorescence (DAPI), which was used as a control procedure. The appearance of the mycoplasma cell border and content gives some information about particle viability.

Simulium (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of São Paulo, Brazil

The larva, pupa, male, and female of Simulium bifenestratum n. sp. are described and illustrated. The pupae of the new species have 10 gill filaments, thick at their base and arranged in a three-dimensional way, surrounding the head and thorax. Its pupal cocoon is peculiar, not found in any of the known Brazilian black-fly species; it is very thick and hard with two openings in the anterior region. S. bifenestratum n. sp. was collected in one stream in the Bocaina mountain chain, Atlantic forest, in São José do Barreiro county, state of São Paulo, in a high (1500 m) natural grassland. Larvae and pupae were collected on the edges of small waterfalls and in places with-high speed laminar water flow, attached to the bedrock.

Sistema automático para determinação seqüencial de cianeto livre e total empregando eletrodo tubular íon-seletivo de membrana homogênea

This study presents an automated system for potentiometric determination of free and total cyanide which employs a homogeneous membrane tubular ion-selective electrode. After the electrode is assembled, it is connected to a system composed of 3 three-way solenoid valves, sample line, carrier line, acid stream, and gas diffusion chamber. A Turbo Pascal® computer program, developed specifically for this task, automatically performs all the steps involved in data acquisition and processing. The proposed analytical procedure offers operational simplicity, since detection is performed by a tubular electrode, whose assembly is fast and easy. The system has shown reproducibility (r.s.d. < 0.5%, n=6) and high speed (30 readings/hour); it is efficient for determination of free and total cyanide in waste waters of starch processing plants. The detection limit was 1.2x10-5 and 1.5x10-5 mol L-1, for determination of free and total cyanide, respectively. The linear response range was between 1.2x10-5 and 1.0x10-2 mol L-1 for free cyanide and between 1.5x10-5 and 1.0x10-2 for total cyanide.