Evaluating the Baylis-Hillman reaction of cyclic enones using surfactants in water

Conjugated cyclic enones react smoothly in water with a variety of aldehydes (Baylis-Hillman reaction) in the presence of surfactants above their critical micelle concentrations (CMC).

HPLC analyses of flavanols and phenolic acids in the fresh young shoots of tea (Camellia sinensis) grown in Australia

As part of a 4-year project to study phenolic compounds in tea shoots over the growing seasons and during black tea processing in Australia, an HPLC method was developed and optimised for the identification and quantification of phenolic compounds, mainly flavanols and phenolic acids, in fresh tea shoots. Methanol proved to be the most suitable solvent for extracting the phenolic compounds, compared with chloroform, ethyl acetate and water. Immediate analysis, by HPLC, of the methanol extract showed higher separation efficiency than analyses after being dried and redissolved. This method exhibited good repeatability (CV 3-9%) and recovery rate (88-116%). Epigallocatechin gallate alone constituted up to 115 mg/g, on a dry basis, in the single sample of Australian fresh tea shoots examined. Four catechins (catechin, gallocatechin, epicatechin and epigallocatechin) and six catechin gallates (epigallocatechin gallate, catechin gallate, epicatechin gallate, gallocatechin gallate, epicatechin digallate and epigallocatechin digallate) have been identified and quantified by this HPLC method. In addition, two major tea alkaloids, caffeine and theobromine, have been quantified, while five flavonol glycosides and six phenolic acids, including quinic acids and esters, were identified and quantified. (C) 2003 Elsevier Ltd. All rights reserved.

Seasonal variations of phenolic compounds in Australia-grown tea (Camellia sinensis)

Seasonal variations of phenolic compounds in fresh tea shoots grown in Australia were studied using an HPLC method. Three principal tea flavanols [epigallocatechin gallate (EGCG), epicatechin gallate (ECG), and epigallocatechin (EGC)] and four grouped phenolics [total catechins (Cs), total catechin gallates (CGs), total flavanols (Fla), and total polyphenols (PPs)] in fresh tea shoots were analyzed and compared during the commercial harvest seasons from April 2000 to May 2001. The levels of EGCG, ECG, and CGs in the fresh tea shoots were higher in the warm months of April 2000 (120.52, 34.50, and 163.75 mg/g, respectively) and May 2000 (128.63, 44.26, and 183.83 mg/g, respectively) and lower during the cool months of July 2000 (91.39, 35.16, and 132.30 mg/g, respectively), August 2000 (91.31, 31.56, and 128.64 mg/g, respectively), and September 2000 (96.12, 33.51, and 136.90 mg/g, respectively). Thereafter, the levels increased throughout the warmer months from October to December 2000 and remained high until May 2001. In the warmer months, the levels of EGCG, ECG, and CGs were in most cases significantly higher (P < 0.05) than those in the samples harvested in the cooler months. In contrast, the levels of EGC and Cs were high and consistent in the cooler months and low in the warmer months. The seasonal variations of the individual and grouped catechins were significant (P < 0.05) between the cooler and warmer months. This study revealed that EGCG and ECG could be used as quality descriptors for monitoring the seasonal variations of phenolics in Australia-grown tea leaves, and the ratio (EGCG + ECG)/EGC has been suggested as a quality index for measuring the differences in flavanol levels in fresh tea shoots across the growing seasons. Mechanisms that induce seasonal variations in tea shoots may include one or all three of the following environmental conditions: day length, sunlight, and/or temperature, which vary markedly across seasons. Therefore, further studies under controlled conditions such as in a greenhouse may be required to direct correlate flavonoid profiles of green tea leaves with their yields and also to with conditions such as rainfall and humidity.

A rapid and sensitive microscale HPLC method for the determination of indomethacin in plasma of premature neonates with patent ductus arteriousus

Indomethacin (IND) is the drug of choice for the closure of a patent ductus arteriosus (PDA) in neonates. This paper describes a simple, sensitive, accurate and precise microscale HPLC method suitable for the analysis of IND in plasma of premature neonates. Samples were prepared by plasma protein precipitation with acetonitrile containing the methyl ester of IND as the internal standard (IS). Chromatography was performed on a Hypersil C-18 column. The mobile phase of methanol, water and orthophosphoric acid (70:29.5:0.5, v/v, respectively), was delivered at 1.5 mL/min and monitored at 270 nm. IND and the IS were eluted at 2.9 and 4.3 min, respectively. Calibrations were linear (r > 0.999) from 25 to 2500 mu g/L. The inter- and intra-day assay imprecision was less than 4.3% at 400-2000 mu g/L, and less than 22.1% at 35 mu g/L. Inaccuracy ranged from -6.0% to +1.0% from 35 to 2000 mu g/L. The absolute recovery of IND over this range was 93.0-113.3%. The IS was stable for at least 36 h when added to plasma at ambient temperature. This method is suitable for pharmacokinetic studies of IND and has potential for monitoring therapy in infants with PDA when a target therapeutic range for IND has been validated. (c) 2005 Elsevier B.V. All rights reserved.

Phosphorus uptake from green manures and phosphate fertilizers applied in an acid tropical soil

Quantifying the relative contribution of different phosphorus (P) sources to P uptake can lead to greater understanding of the mechanisms that increase available P in integrated P management systems. The P-32-P-33 double isotope labeling technique was used to determine the relative contribution of green manures (GMs) and P fertilizers to P uptake by Setaria grass (Setaria sphacelata) grown in an amended tropical acid soil (Bungor series) in a glasshouse study. The amendments were factorial combinations of GMs (Calopogonium caeruleum , Gliricidia sepium and Imperata cylindrica) and P fertilizers [phosphate rocks (PRs) from North Carolina (NCPR), China (CPR) and Algeria (APR), and triple superphosphate (TSP)]. Dry matter yield, P uptake, and P utilization from the amendments were monitored at 4, 8, and 15 weeks after establishment (WAE). The GMs alone or in combination with P fertilizers contributed less than 5% to total P uptake in this soil, but total P uptake into Setaria plants in the GM treatments was three to four times that of the P fertilizers because the GMs mobilized more soil P. Also, the GMs markedly increased fertilizer P utilization in the combined treatments, from 3% to 39% with CPR, from 6-9% to 19-48% with reactive PRs, and from 6% to 37% with TSP in this soil. Both P GM and the other decomposition products were probably involved in reducing soil P-retention capacity. Mobilization of soil P was most likely the result of the action of the other decomposition products. These results demonstrate the high potential of integrating GMs and PRs for managing P in tropical soils and the importance of the soil P mobilization capacity of the organic components. Even the low-quality Imperata GM enhanced the effectiveness of the reactive APR more than fourfold.

RP-HPLC detection of a sulphiting-induced artefact from paeoniflorin in dried roots of Paeonia lactiflora

Paeoniflorin is one of the bioactive ingredients of the roots of Paeonia lactiflora (Paeoniaceae). A comparative study of processed and non-processed commercial samples of dried roots of P. lactiflora indicated a very low level of paeoniflorin in the processed sample and the formation of a new more polar component, sodium paeoniflorin sulphonate, during treatment of the roots with sulphiting agents. Copyright (c) 2006 John Wiley & Sons, Ltd.

Taxonomy of Australian Funnel-web spiders using rp-HPLC/ESI-MS profiling techniques

The complex nature of venom from spider species offers a unique natural source of potential pharmacological tools and therapeutic leads. The increased interest in spider venom molecules requires reproducible and precise identification methods. The current taxonomy of the Australian Funnel-web spiders is incomplete, and therefore, accurate identification of these spiders is difficult. Here, we present a study of venom from numerous morphologically similar specimens of the Hadronyche infensa species group collected from a variety of geographic locations in southeast Queensland. Analysis of the crude venoms using online reversed-phase high performance liquid chromatography/electrospray ionisation mass spectrometry (rp-HPLC/ESI-MS) revealed that the venom profiles provide a useful means of specimen identification, from the species level to species variants. Tables defining the descriptor molecules for each group of specimens were constructed and provided a quick reference of the relationship between one specimen and another. The study revealed that the morphologically similar specimens from the southeast Queensland region are a number of different species/species variants. Furthermore, the study supports aspects of the current taxonomy with respect to the H. infensa species group. Analysis of Australian Funnel-web spider venom by rp-HPLC/ESI-MS provides a rapid and accurate method of species/species variant identification. (c) 2006 Elsevier Ltd. All rights reserved.