Investigating the Protective Role of Biochaga Drug on Structural Changes of Bovine Serum Albumin in the Presence of Methyl tert-butyl Ether.

BACKGROUND: The health benefits of natural products have a long history. Chaga (Inonotus obliques) is used in traditional medicine and is an essential antioxidant for protecting the body from oxidants. Reactive oxygen species (ROS) are produced routinely due to metabolic processes. However, environmental pollution factors such as methyl tert-butyl ether (MTBE) can increase oxidative stress in the human body. MTBE is widely used as a fuel oxygenator that can harm health. The widespread use of MTBE has posed significant threats to the environment by polluting environmental resources, including groundwater. This compound can accumulate in the bloodstream by inhaling polluted air, with a strong affinity for blood proteins. The primary mechanism of MTBE's harmful effects is ROS production. The use of antioxidants may help reduce MTBE oxidation conditions. The present study proposes that biochaga, as an antioxidant, can reduce MTBE damage in the bovine serum albumin (BSA) structure. METHODS AND RESULTS: This study investigated the role of different concentrations of biochaga in the structural change of BSA in the presence of MTBE by biophysical methods such as UV-Vis, fluorescence, FTIR spectroscopy, DPPH radical inhibition method, aggregation test, and molecular docking. Research at the molecular level is critical to investigate the structural change of proteins by MTBE and the protective effect of the ideal dose (2.5 µg/ml) of biochaga. CONCLUSION: the results of spectroscopic examinations showed that the concentration of 2.5 µg/ml of biochaga has the least destructive effect on the structure of BSA in the presence and absence of MTBE, and it can play as an antioxidant.

Bioremediation of gaseous methyl tert-butyl ether by combination of sulfuric acid modified bagasse activated carbon-bone biochar beads and Acinetobacter indicus screened from petroleum contaminated soil.

Combination of sulfuric acid modified bagasse activated carbon-bone biochar beads and Acinetobacter indicus screened from petroleum contaminated soil was the best condition for gaseous methyl tert-butyl ether (MTBE) removal. It was found that H2SO4 modified bagasse AC in powder form had higher adsorption capacity (989.33 mg g-1) than that in bead form (1.94 mg g-1). In addition, bone biochar in powder form (3.51 mg g-1) also had higher adsorption capacity than that in bead form (1.63 mg g-1). This was the fact that material beads contained high moisture content that inhibited the penetration of gaseous MTBE into the material. And a mixed material of H2SO4 modified bagasse AC-bone biochar beads had the highest adsorption capacity (2.22 mg g-1) compared to individual H2SO4 modified bagasse AC beads (1.94 mg g-1) and bone biochar beads (1.63 mg g-1) due to a mixed material had more rough surface and high surface area on its material. So, gaseous MTBE can penetrate through this material more easily. Although the maximum adsorption capacity of H2SO4 modified bagasse AC in powder form was the highest but microorganism cannot sustain and survive in this form for a long time. Therefore, the material beads were more suitable for microorganism to grow and degrade gaseous MTBE. Microorganism can degrade MTBE and caused no secondary wastes. Moreover, A. indicus was a novel strain for MTBE removal that has not been previously reported. Therefore, a combination of A. indicus-mixed material beads was a good choice for MTBE removal in a biofilter system.

Purification, crystallization and anticancer activity evaluation of the compound alternariol methyl ether from endophytic fungi Alternaria alternata.

AIMS: Medicinal plant-associated endophytic fungi are important sources of precious bioactive compounds, contributing more than 80% of the natural drugs for various ailments. The present study was aimed at evaluating the anticancer activity of the crystallized compound alternariol methyl ether (AME) against hepatocellular carcinoma (HCC) both in vitro and in vivo from an endophytic fungus residing in the medicinal plant Vitex negundo. METHODS AND RESULTS: The secondary metabolites from the endophytic fungus Alternaria alternata MGTMMP031 were isolated. Purification and characterization of the compound was performed and the potential compound was identified as AME. The crystal structure of AME was unambiguously confirmed by X-ray analysis. AME has been checked for its antibacterial and anticancer properties which showed its effectiveness against various bacteria and demonstrated marked anti-proliferative activity against the human HCC cells (HUH-7) both in vitro and in vivo. Mode of actions included cell cycle arrest, reducing the level of markers enzymes of liver cancer and preventing tumour growth. CONCLUSIONS: Alternariol methyl ether acts as a potential therapeutic target against HCC. The compound was isolated and the crystal structure was obtained for the first time from the endophytic fungus A. alternata MGTMMP031. In the present study, the crystallized structure of AME was obtained by slow evaporation technique. It can be concluded that AME acts as a potential therapeutic target against HCC. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic fungi residing in the medicinal plants have strong biological significance and bioactive compounds from these fungi provide better therapeutic targets against diseases.

Use of omic tools to assess methyl tert-butyl ether (MTBE) degradation in groundwater.

This study employed innovative technologies to evaluate multiple lines of evidence for natural attenuation (NA) of methyl tertiary-butyl ether (MTBE) in groundwater at the 22 Area of Marine Corps Base (MCB) Camp Pendleton after decommissioning of a biobarrier system. For comparison, data from the 13 Area Gas Station where active treatment of MTBE is occurring was used to evaluate the effectiveness of omic techniques in assessing biodegradation. Overall, the 22 Area Gas Station appeared to be anoxic. MTBE was detected in large portion of the plume. In comparison, concentrations of MTBE at the 13 Area Gas Station were much higher (42,000 µg/L to 2800 µg/L); however, none of the oxygenates were detected. Metagenomic analysis of the indigenous groundwater microbial community revealed the presence of bacterial strains known to aerobically and anaerobically degrade MTBE at both sites. While proteomic analysis at the 22 Area Gas Station showed the presence of proteins of MTBE degrading microorganisms, the MTBE degradative proteins were only found at the 13 Area Gas Station. Taken together, these results provide evidence for previous NA of MTBE in the groundwater at 22 Area Gas Station and demonstrate the effectiveness of innovative-omic technologies to assist monitored NA assessments.

Selective Inhibition of Human Monoamine Oxidase B by Acacetin 7-Methyl Ether Isolated from Turnera diffusa (Damiana).

The investigation of the constituents that were isolated from Turnera diffusa (damiana) for their inhibitory activities against recombinant human monoamine oxidases (MAO-A and MAO-B) in vitro identified acacetin 7-methyl ether as a potent selective inhibitor of MAO-B (IC50 = 198 nM). Acacetin 7-methyl ether (also known as 5-hydroxy-4', 7-dimethoxyflavone) is a naturally occurring flavone that is present in many plants and vegetables. Acacetin 7-methyl ether was four-fold less potent as an inhibitor of MAO-B when compared to acacetin (IC50 = 50 nM). However, acacetin 7-methyl ether was >500-fold selective against MAO-B over MAO-A as compared to only two-fold selectivity shown by acacetin. Even though the IC50 for inhibition of MAO-B by acacetin 7-methyl ether was ~four-fold higher than that of the standard drug deprenyl (i.e., SelegilineTM or ZelaparTM, a selective MAO-B inhibitor), acacetin 7-methyl ether's selectivity for MAO-B over MAO-A inhibition was greater than that of deprenyl (>500- vs. 450-fold). The binding of acacetin 7-methyl ether to MAO-B was reversible and time-independent, as revealed by enzyme-inhibitor complex equilibrium dialysis assays. The investigation on the enzyme inhibition-kinetics analysis with varying concentrations of acacetin 7-methyl ether and the substrate (kynuramine) suggested a competitive mechanism of inhibition of MAO-B by acacetin 7-methyl ether with Ki value of 45 nM. The docking scores and binding-free energies of acacetin 7-methyl ether to the X-ray crystal structures of MAO-A and MAO-B confirmed the selectivity of binding of this molecule to MAO-B over MAO-A. In addition, molecular dynamics results also revealed that acacetin 7-methyl ether formed a stable and strong complex with MAO-B. The selective inhibition of MAO-B suggests further investigations on acacetin 7-methyl as a potential new drug lead for the treatment of neurodegenerative disorders, including Parkinson's disease.

Biochemical significance of limonene and its metabolites: future prospects for designing and developing highly potent anticancer drugs.

Monocyclic monoterpenes have been recognized as useful pharmacological ingredients due to their ability to treat numerous diseases. Limonene and perillyl alcohol as well as their metabolites (especially perillic acid and its methyl ester) possess bioactivities such as antitumor, antiviral, anti-inflammatory, and antibacterial agents. These therapeutic properties have been well documented. Based on the aforementioned biological properties of limonene and its metabolites, their structural modification and development into effective drugs could be rewarding. However, utilization of these monocyclic monoterpenes as scaffolds for the design and developments of more effective chemoprotective agents has not received the needed attention by medicinal scientists. Recently, some derivatives of limonene metabolites have been synthesized. Nonetheless, there have been no thorough studies on their pharmacokinetic and pharmacodynamic properties as well as their inhibition against isoprenylation enzymes. In this review, recent research progress in the biochemical significance of limonene and its metabolites was summarized with emphasis on their antitumor effects. Future prospects of these bioactive monoterpenes for drug design and development are also highlighted.

NOx-smoke trade-off characteristics of minor vegetable oil blends synergy with oxygenate in a commercial CI engine.

The present study investigates the effect of blending oxygenate namely diethylene glycol dimethyl ether (diglyme) with minor vegetable oil namely rubber seed oil (RSO), babassu oil (BSO), and their blends in various proportions (R75B25, R50B50, and R25B75) on NOx-smoke trade-off and other engine characteristics. The tests were conducted on a commercial twin cylinder compression-ignition (CI) engine commonly used in tractors. The potential of the blends with diglyme is assessed based on performance, emission, and combustion characteristics of the engine at different load conditions. The tests were conducted at a constant speed of 1500 rpm maintaining the original injection timing and pressure. Compared to diesel, RSO, and BSO, and their blends exhibited inferior combustion due to poor physical properties like high viscosity and density. This resulted in a lower brake thermal efficiency with increase in HC, CO, and smoke emissions compared to diesel at all the load conditions. The augmented effect is observed with increase in BSO proportion for the blends and neat BSO. The poor combustion of minor vegetable oil and its blends lead to lower NOx emission as a result of lower in-cylinder temperature. To improve the performance and NOx-smoke trade-off, diglyme (DGM) was added with all the test fuels with the optimum share of 20% (by volume). Addition of DGM, increased brake thermal efficiency by 2-7% for all the test fuels due to improved combustion as a result of additional fuel bound oxygen in DGM and improved fuel blend properties. DGM addition reduced smoke, HC, and CO emission drastically with a slight increase in NOx emission compared to minor vegetable oil blends. The study shows that addition of DGM showed a promising note in NOx-smoke trade-off without affecting the other engine parameters.

Discovery of potent and selective acetylcholinesterase (AChE) inhibitors: acacetin 7-O-methyl ether Mannich base derivatives synthesised from easy access natural product naringin.

Naringin, as a component universal existing in the peel of some fruits or medicinal plants, was usually selected as the material to synthesise bioactive derivates since it was easy to gain with low cost. In present investigation, eight new acacetin-7-O-methyl ether Mannich base derivatives (1-8) were synthesised from naringin. The bioactivity evaluation revealed that most of them exhibited moderate or potent acetylcholinesterase (AChE) inhibitory activity. Among them, compound 7 (IC50 for AChE = 0.82 ± 0.08 µmol•L-1, IC50 for BuChE = 46.30 ± 3.26 µmol•L-1) showed a potent activity and high selectivity compared with the positive control Rivastigmine (IC50 for AChE = 10.54 ± 0.86 µmol•L-1, IC50 for BuChE = 0.26 ± 0.08 µmol•L-1). The kinetic study suggested that compound 7 bind to AChE with mix-type inhibitory profile. Molecular docking study revealed that compound 7 could combine both catalytic active site (CAS) and peripheral active site (PAS) of AChE with four points (Trp84, Trp279, Tyr70 and Phe330), while it could bind with BuChE via only His 20.

(R)-(-)-Aloesaponol III 8-Methyl Ether from Eremurus persicus: A Novel Compound against Leishmaniosis.

Leishmaniosis is a neglected tropical disease which affects several millions of people worldwide. The current drug therapies are expensive and often lack efficacy, mainly due to the development of parasite resistance. Hence, there is an urgent need for new drugs effective against Leishmania infections. As a part of our ongoing study on the phytochemical characterization and biological investigation of plants used in the traditional medicine of western and central Asia, in the present study, we focused on Eremurus persicus root extract in order to evaluate its potential in the treatment of leishmaniosis. As a result of our study, aloesaponol III 8-methyl ether (ASME) was isolated for the first time from Eremurus persicus root extract, its chemical structure elucidated by means of IR and NMR experiments and the (R) configuration assigned by optical activity measurements: chiroptical aspects were investigated with vibrational circular dichroism (VCD) and electronic circular dichroism (ECD) spectroscopies and DFT (density functional theory) quantum mechanical calculations. Concerning biological investigations, our results clearly proved that (R)-ASME inhibits Leishmania infantum promastigotes viability (IC50 73 µg/mL), inducing morphological alterations and mitochondrial potential deregulation. Moreover, it is not toxic on macrophages at the concentration tested, thus representing a promising molecule against Leishmania infections.

Fatty acid composition of achenes of Cirsium taxa (Asteraceae, Carduoideae) from Turkey.

The fatty acid compositions and total oil contents in achenes of 22 Cirsium taxa collected from different natural habitats in Turkey were investigated. The range of total fat in the taxa varied from 1.7% to 13.3%. The taxa contained palmitic (C16:0), stearic (C18:0), oleic (C18:1n-9), linoleic (C18:2n-6), and α-linolenic (C18:3n-3) acid. Polyunsaturated fatty acids had the highest level, ranging from 52.1% to 75.2% (C18:2n-6) and 0.5%-17.3% (C18:3n-3). Total saturated (6.9%-17.4%), monounsaturated (11.7%-28.9%), and polyunsaturated (57.4%-79.9%) fatty acids varied substantially, whereas total unsaturated fatty acids ranged from 70.6% to 91.3%. Considerable variation was observed (P<0.05) within each taxon for these fatty acids, especially α-linolenic acid (C18:3n-3). Fatty acid compositions of the 22 taxa were compared by cluster analysis (UPGMA) and principle component analysis (PCA) based on six major fatty acids. This is the first report on the lipid content and fatty acid composition of achenes of Cirsium taxa growing in different areas of Turkey. Fatty acid profiles, relative proportions and levels of fatty acids can be used as additional biochemical markers in the taxonomy of Cirsium.

The Effectiveness and Stability of a 20% Emulsified Sevoflurane Formulation for Intravenous Use in Rats.

BACKGROUND: Halogenated volatile anesthetics can be safely and rapidly administered to animals and humans using emulsion formulations. However, they must be administered simultaneously with a high dose of lipids. Increasing the concentration of volatile anesthetics may solve this clinical issue. Moreover, careful observation is needed when the emulsion is injected because anaphylactic reactions have been reported. METHODS: We prepared a 20% sevoflurane lipid emulsion and administered it to 69 male Sprague-Dawley rats via the tail vein. The median effective dose (ED50) for the loss of righting reflex and the median lethal dose (LD50) were determined. ED50 and LD50 values were calculated using nonlinear regression, and data were fitted with a cumulative Gaussian model using GraphPad Prism. Measurements of vital signs and evaluation of the presence of adverse effects associated with continuous infusion of emulsions were verified. Stability of the emulsion was assessed by measuring particle size at 365 days and sevoflurane concentrations after opening the vial at 180 minutes. RESULTS: The ED50 and LD50 were 0.47 mL/kg (95% confidence interval [CI], 0.46-0.48) and 1.13 mL/kg (95% CI, 1.07-1.18), respectively. The therapeutic index (LD50/ED50) was 2.41 (95 CI%, 2.23-2.59), which compares favorably with therapeutic index of a fluoropolymer-based emulsion of sevoflurane, propofol, and thiopental. There were no adverse effects associated with the continuous infusion of emulsions. Particle size of the emulsion at 365 days after preparation was 78.9 ± 3.8 nm (±SD), and sevoflurane concentration at 180 minutes after opening the vial was 19.0% ± 0.6% (±SD). CONCLUSIONS: We prepared a 20% sevoflurane lipid emulsion using caprylic triglyceride (i.e., medium-chain triglyceride). In rats, this emulsion was an effective anesthetic and was not associated with adverse events. The emulsion was stable after consecutive evaluation for 365 days and for 180 minutes after the vial was opened.

The Isolation of Lutein and Lutein 3'-methyl ether from Peristrophe lanceolaria.

Two carotenoids, lutein (1) and lutein 3'-methyl ether (2), have been isolated from the EtOAc fraction othe MeOH extract of Peristrophe lanceolaria, growing in Thailand. The structures of these compounds were elucidated from their ID and 2D NMR spectroscopic data and from comparisons made with the literature data. This is the first report of the isolation of lutein-3'-methyl ether as a natural product.

Development of a method to screen and isolate potential α-glucosidase inhibitors from Panax japonicus C.A. Meyer by ultrafiltration, liquid chromatography, and counter-current chromatography.

A new assay based on ultrafiltration, liquid chromatography and mass spectrometry was developed for the rapid screening and identification of the ligands for α-glucosidase from the extract of Panax japonicus. Six saponins were identified as α-glucosidase inhibitors. Subsequently, the specific binding ligands, namely, notoginsenoside R1 , ginsenoside Rb1 , chikusetsusaponin V, chikusetsusaponin IV, chikusetsusaponin IVa, and ginsenoside Rd (the purities were 94.18, 95.43, 96.09, 93.26, 94.50, 93.86%, respectively) were separated by counter-current chromatography using two-phase solvent systems composed of tert-butyl methyl ether, acetonitrile, 0.1% aqueous formic acid (3.8:1.0:4.4, v/v/v) and the solvent system composed of methylene chloride, isopropanol, methanol, 0.1% aqueous formic acid (5.8:1.0:6.0:2.2, v/v/v). The results demonstrate that ultrafiltration, liquid chromatography and mass spectrometry combined with high-speed counter-current chromatography might provide not only a powerful tool for screening and isolating α-glucosidase inhibitors in complex samples but also a useful platform for discovering bioactive compounds for the prevention and treatment of diabetes mellitus.

Three-phase solvent systems for the comprehensive separation of a wide variety of compounds from Dicranostigma leptopodum by high-speed counter-current chromatography.

A three-phase solvent system was efficiently applied for high-speed counter-current chromatography to separate secondary metabolites with a wide range of hydrophobicity in Dicranostigma leptopodum. The three-phase solvent system of n-hexane/methyl tert-butyl ether/acetonitrile/0.5% triethylamine (2:2:3:2, v/v/v/v) was selected for high-speed counter-current chromatography separation. The separation was initiated by filling the column with a mixture of intermediate phase and lower phase as a stationary phase followed by elution with upper phase to separate the hydrophobic compounds. Then the mobile phase was switched to the intermediate phase to elute the moderately hydrophobic compounds, and finally the polar compounds still retained in the column were fractionated by eluting the column with the lower phase. In this research, 12 peaks were eluted out in one-step operation within 110 min, among them, eight compounds with acceptable purity were obtained and identified. The purities of ß-sitosterol, protopine, allocryptopine, isocorydione, isocorydine, coptisine, berberrubine, and berberine were 94.7, 96.5, 97.9, 86.6, 98.9, 97.6, 95.7, and 92.8%, respectively.

Simultaneous determination of seven taxoids in rat plasma by UPLC-MS/MS and pharmacokinetic study after oral administration of Taxus yunnanensis extracts.

A rapid, sensitive and reliable method has been developed and validated for the simultaneous determination of seven taxoids including 10-deacetylbaccatin III (10-DAB III), baccatin III, 5-epi-canadensene, taxinine M, 10-deacetyltaxol (10-DAT), cephalomannine and paclitaxel in rat plasma using docetaxel as the internal standard (IS). The plasma samples were pretreated by liquid-liquid extraction with methyl tert-butyl ether. The chromatographic separation was achieved on a C18 column (50 mm × 2.1 mm, 1.8 µm, Waters, USA) with a gradient elution program consisting of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. Detection was performed under the selected reaction monitoring (SRM) scan using an electrospray ionization (ESI) in the positive ion mode. The mass transitions were as follows: m/z 567.4→444.9 for 10-DAB III, m/z 609.0→549.3 for baccatin III, m/z 617.4→496.9 for 5-epi-canadensene, m/z 709.6→649.3 for taxinine M, m/z 834.8→307.9 for 10-DAT, m/z 854.5→285.4 for cephalomannine, m/z 876.8→307.3 for paclitaxel and m/z 830.8→549.6 for IS, respectively. All calibration curves exhibited good linearity (r(2)>0.99) over a wide concentration range for all components. The intra-day and inter-day precisions at three different levels were both less than 14.3% in terms of relative standard deviation (RSD) and the accuracies ranged from -8.3% to 14.8% in terms of relative error (RE). The extraction recoveries of the seven compounds ranged from 62.5% to 100.5%. The developed method was successfully applied to the pharmacokinetic study of the seven taxoids in rat plasma after oral administration of the crude extract of the twigs and leaves of Taxus yunnanensis.

Reduction of diffusive contaminant emissions from a dissolved source in a lower permeability layer by sodium persulfate treatment.

Residual contamination contained in lower permeability zones is difficult to remediate and can, through diffusive emissions to adjacent higher permeability zones, result in long-term impacts to groundwater. This work investigated the effectiveness of oxidant delivery for reducing diffusive emissions from lower permeability zones. The experiment was conducted in a 1.2 m tall × 1.2 m wide × 6 cm thick tank containing two soil layers having 3 orders of magnitude contrast in hydraulic conductivity. The lower permeability layer initially contained dissolved methyl tert-butyl ether (MTBE) and benzene, toluene, ethylbenzene, and p-xylenes (BTEX). The treatment involved delivery of 10% w/w nonactivated sodium persulfate (Na2S2O8) solution to the high permeability layer for 14 days. The subsequent diffusion into the lower permeability layer and contaminant emission response were monitored for about 240 days. The S2O8(2-) diffused about 14 cm at 1% w/w into the lower permeability layer during the 14 day delivery and continued diffusing deeper into the layer as well as back toward the higher-lower permeability interface after delivery ceased. Over 209 days, the S2O8(2-) diffused 60 cm into the lower permeability layer, the BTEX mass and emission rate were reduced by 95-99%, and the MTBE emission rate was reduced by 63%. The overall treatment efficiency was about 60-110 g-S2O8(2-)delivered/g-hydrocarbon oxidized, with a significant fraction of the oxidant delivered likely lost by back-diffusion and not involved in hydrocarbon destruction.

Separation of nine compounds from Salvia plebeia R.Br. using two-step high-speed counter-current chromatography with different elution modes.

Nine compounds were successfully separated from Salvia plebeia R.Br. using two-step high-speed counter-current chromatography with three elution modes. Elution-extrusion counter-current chromatography was applied in the first step, while classical counter-current chromatography and recycling counter-current chromatography were used in the second step. Three solvent systems, n-hexane/ethyl acetate/ethanol/water (4:6.5:3:7, v/v), methyl tert-butyl ether/ethyl acetate/n-butanol/methanol/water (6:4:1:2:8, v/v) and n-hexane/ethyl acetate/methanol/water (5:5.5:5:5, v/v) were screened and optimized for the two-step separation. The separation yielded nine compounds, including caffeic acid (1), 6-hydroxyluteuolin-7-glucoside (2), 5,7,3',4'-tetrahydroxy-6-methoxyflavanone-7-glucoside (3), nepitrin (4), rosmarinic acid (5), homoplantaginin (6), nepetin (7), hispidulin (8), and 5,6,7,4'-tertrahydroxyflavone (9). To the best of our knowledge, 5,7,3',4'-tetrahydroxy-6-methoxyflavanone-7-glucoside and 5,6,7,4'-tertrahydroxyflavone have been separated from Salvia plebeia R.Br. for the first time. The purities and structures of these compounds were identified by high-performance liquid chromatography, electrospray ionization mass spectrometry, (1)H and (13)C NMR spectroscopy. This study demonstrates that high-speed counter-current chromatography is a useful and flexible tool for the separation of components from a complex sample.

[Simultaneous determination of seven constituents in Lamiophlomis rotata by HPLC].

OBJECTIVE: To establish a new method for simultaneous determination of shanzhiside methyl ester, chlorogenic acid, 8-O-acetyl shanzhiside methylester, forsythiaside B, rutin, acteoside and galuteolin in Lamiophlomis rotata. METHOD: Separation was performed on a Welchrom-C18 chromatographic column with acetonitrile-0.1% orthophosphoric acid as mobile phasewith gradient elution. The flow rate was 1.0 mL x min(-1). The column temperature was 30 degrees C, and the detection wavelength was set at 238 nm, 330 nm and 350 nm. RESULT: The seven compounds were well separated with good linear correlations. The mean recoveries of seven compounds were 96.47%-102.2% (RSD 0.70%-2.2%). CONCLUSION: There were good correlations among the seven compounds in the samples of aerial parts. The mean sum of shanzhiside methyl ester and 8-O-acetyl shanzhiside methylester in samples of aerial parts is 1.44%. The aerial parts have more kinds of composition and with higher content than that of underground parts in L. rotata, which was reasonable for the resonable use of the aerial part as medicinal part. The method was simple, repeatable and stable, which could be used for identification and quality evaluation of L. rotata.

Direct LSC method for measurements of biofuels in fuel.

Direct liquid scintillation counting (LSC) for quantification of biofuels content in fuels was implemented and validated on three liquid fossil fuel matrices-ethanol, gasoline and diesel. Fatty acid methyl esters (FAMEs), hydrogenated vegetable oils (HVO) and bio-ethanol were used as biofuels. The method is applicable in the range up to 100% for all tested combinations of bio components. The sensitivity and precision of the method are suitable for determination of bio component content in the blends which is appearing on the global market. The method does not require special equipment for sample preparation.

Stepwise elution of a three-phase solvent system in centrifugal partition extraction: a new strategy for the fractionation and phytochemical screening of a crude bark extract.

INTRODUCTION: Tree bark represents an interesting source of bioactive molecules for the discovery of new pharmaceutical agents. However, the detailed screening of secondary metabolites in crude bark extracts is often hampered by the presence of tannins, which are difficult to separate from other plant constituents. OBJECTIVE: In the present study, a new centrifugal partition extraction (CPE) method was developed in order to fractionate a crude bark extract of Anogeissus leiocarpus Guill. & Perr. (Combretaceae). METHODS: A three-phase solvent system composed of n-heptane, methyl tert-butyl ether, acetonitrile and water was optimised for the stepwise elution at 20 mL/min of different phytochemical classes according to their hydrophobicity. Onedimensional and two-dimensional NMR analyses of the simplified fractions were then performed in order to characterise potentially interesting metabolites. RESULTS: In one step, 5 g of the initial crude extract were efficiently fractionated to yield highly simplified fractions that contained triterpenes, ellagic acid derivatives, flavonoids and phenolic compounds. All undesired compounds, that is, the highly abundant water-soluble tannins (78.8%), were totally removed and each run was rapidly achieved in 90 min on a the multi-gram scale and with low solvent volumes. CONCLUSION: Centrifugal partition extraction in the elution mode using a three-phase solvent system can thus be proposed as an efficient and cost-effective alternative for a rapid fractionation of crude bark extracts and for an effective screening of potentially active secondary metabolites.