Introduction: The genus Clusia L. is mostly recognised for the production of prenylated benzophenones and tocotrienol derivatives.Objectives: The objective of this study was to map metabolome variation within Clusia minor organs at different developmental stages.Material and Methods: In total 15 organs/stages (leaf, flower, fruit, and seed) were analysed by UPLC‐MS and 1H‐ and heteronuclear multiple‐bond correlation (HMBC)‐NMR‐based metabolomics.Results: This work led to the assignment of 46 metabolites, belonging to organic acids(1), sugars(2) phenolic acids(1), flavonoids(3) prenylated xanthones(1) benzophenones(4) and tocotrienols(2). Multivariate data analyses explained the variability and classification of samples, highlighting chemical markers that discriminate each organ/stage. Leaves were found to be rich in 5‐hydroxy‐8‐methyltocotrienol (8.5 μg/mg f.w.), while flowers were abundant in the polyprenylated benzophenone nemorosone with maximum level detected in the fully mature flower bud (43 μg/mg f.w.). Nemorosone and 5‐hydroxy tocotrienoloic acid were isolated from FL6 for full structural characterisation. This is the first report of the NMR assignments of 5‐hydroxy tocotrienoloic acid, and its maximum level was detected in the mature fruit at 50 μg/mg f.w. Seeds as typical storage organ were rich in sugars and omega‐6 fatty acids.Conclusion: To the best of our knowledge, this is the first report on a comparative 1D‐/2D‐NMR approach to assess compositional differences in ontogeny studies compared with LC‐MS exemplified by Clusia organs. Results derived from this study provide better understanding of the stages at which maximal production of natural compounds occur and elucidate in which developmental stages the enzymes responsible for the production of such metabolites are preferentially expressed.

Amino acids (AAs), important constituents of natural moisturizing factors (NMFs) of the skin are decreased in diseased conditions such as psoriasis and atopic dermatitis. No study so far investigated the uptake of AAs into isolated corneocytes (COR). The present study was performed using 19 AAs, including taurine (TAU), to measure their amount diffused into the COR and binding of these AAs to keratin. Incubation of alanine, aspartic acid, asparagine, glutamine, glutamic acid, histidine, proline, serine and TAU with the isolated COR showed uptake after 24 h of 51.6, 95.4, 98.6, 94.1, 95.6, 90.1, 94.6, 72.9 and 57.8 %, respectively, into the COR but no binding with keratin. Uptake of TAU was validated by time dependent in-vitro diffusion models 'without COR and 'with COR'. The time dependent curve fitting showed that in in-vitro diffusion model 'without COR' there was no change in the total concentration of TAU until 72 hours, while in diffusion model 'with COR' the total conc. decreased to 37.8 % after 72 hours. The Pearson's correlation coefficient 'r' between the conc. curves of both in-vitro diffusion models was -0.54 that was an evidence of significant amount of TAU uptake by the COR. AAs as part of the NMFs have a great potential to be diffused into the COR. This property of the AAs can be employed in further dermatological research on diseased or aged skin conditions with NMFs deficiency.

Hydnora abyssinica A.Br. (Hydnoraceae), a holoparasitic herb, is for the first time recorded for Abyan governorate of South Yemen. Flowers of this species were studied for their ethnobotanical, biological and chemical properties for the first time. In South Yemen, they are traditionally used as wild food and to cure stomach diseases, gastric ulcer and cancer. Phytochemical analysis of the extracts showed the presence of terpenes, tannins, phenols, and flavonoids. The volatile components of the air-dried powdered flowers were identified using a static headspace GC/MS analysis as acetic acid, ethyl acetate, sabinene, α-terpinene, (+)-D-limonene and γ-terpinene. These volatile compounds that characterize the odor and taste of the flowers were detected for the first time in a species of the family Hydnoraceae. The flowers were extracted by n-hexane, dichlormethane, ethyl acetate, ethanol, methanol and water. With exception of the water extract all extracts demonstrated activities against Gram-positive bacteria as well as remarkable radical scavenging activities in DPPH assay. Ethyl acetate, methanol and water extracts exhibited good antifungal activities. The cytotoxic activity of the extracts against FL cells, measured in neutral red assay, was only weak (IC50 > 500 μg /mL). The results justify the traditional use of the flowers of Hydnora abyssinica in South Yemen.

The β ‐carboline alkaloids harmane (1 ) and norharmane (2 ) were isolated from fruiting bodies of Hygrophorus eburneus (Bull.) Fr. as well as brunnein A (3 ) from Hygrophorus hyacinthinus Quél. (Tricholomataceae, Agaricales) for the first time. Their occurrence within the genus was investigated using liquid chromatography/electrospray ionisation tandem mass spectrometric methods, especially by selected reaction monitoring. Based on these results their chemotaxonomical relevance is discussed.

Chemical investigation of Stephania rotunda Lour. growing in Viet Nam led to the isolation and structural elucidation of three new alkaloids, 5-hydroxy-6,7-dimethoxy-3,4-dihydroisoquinolin-1(2H)-one (1), thaicanine 4-O-β-L-glucoside (6), as well as (–)-thaicanine N-oxide (4-hydroxycorynoxidine) (8), along with 23 known alkaloids. These structures were determined on the basis of MS and NMR spectroscopic data.

A simple method involving polyamide column chromatography in combination with HPLC‐PAD and HPLC‐ESI[sol ]MS for isolating and identifying two kinds of lignans, arctiin and arctigenin, in the leaves of burdock (Arctium lappa L.) has been established. After extraction of burdock leaves with 80% methanol, the aqueous phase of crude extracts was partitioned between water and chloroform and the aqueous phase was fractionated on a polyamide glass column. The fraction, eluting with 100% methanol, was concentrated and gave a white precipitate at 4°C from which two main compounds were purified by semi‐preparative HPLC. In comparison with the UV and ESI‐MS spectra and the HPLC retention time of authentic standards, the compounds were determined to be arctiin and arctigenin. The extraction[sol ]separation technique was validated using an internal standard method. Copyright © 2005 John Wiley & Sons, Ltd.

2,4-Dimethoxy-2-methyl-6H-pyran-3-one (1), a hitherto unknown natural product, and the calcium salt of rehmapicroside (2) have been isolated from rhizomes of the Vietnamese variety of Rehmannia glutinosa Libosch together with a series of known compounds: norcarotenoids (3–5), 2-formyl-5-hydroxymethylfurane (6), the iridoid rehmaglutin D (7), iridoid glycosides (8–12) and phenylethyl alcohol glycosides (13–17). Their structures were determined by mass and NMR spectroscopy.

Natural isothiocyanates, produced during plant tissue damage from methionine‐derived glucosinolates, are potent inducers of mammalian phase 2 detoxification enzymes such as quinone reductase (QR). A greatly simplified bioassay for glucosinolates based on induction and colorimetric detection of QR activity in murine hepatoma cells is described. It is demonstrated that excised leaf disks of Arabidopsis thaliana (ecotype Columbia) can directly and reproducibly substitute for cell‐free leaf extracts as inducers of murine QR, which reduces sample preparation to a minimum and maximizes throughput. A comparison of 1 and 3 mm diameter leaf disks indicated that QR inducer potency was proportional to disk circumference (extent of tissue damage) rather than to area. When compared to the QR inducer potency of the corresponding amount of extract, 1 mm leaf disks were equally effective, whereas 3 mm disks were 70% as potent. The QR inducer potency of leaf disks correlated positively with the content of methionine‐derived glucosinolates, as shown by the analysis of wild‐type plants and mutant lines with lower or higher glucosinolate content. Thus, the microtitre plate‐based assay of single leaf disks provides a robust and inexpensive visual method for rapidly screening large numbers of plants in mapping populations or mutant collections and may be applicable to other glucosinolate‐producing species.

After application of [17‐D2]‐GA34 to seedlings of Phaseolus coccineus L. cv. Prizewinner, both metabolically formed conjugates [17‐D2]‐GA34‐2‐O‐glucoside and [17‐D2]‐GA34‐glucosyl ester could be established structurally. The identification was based on data from GC‐MS as well as LC‐ESI‐tandem MS (negative ions) techniques (daughter ion scan, parent ion scan, selected ion monitoring, selected reaction monitoring), and NMR studies.

Conjugates of 3,7‐trans cucurbic acid isomers with either (S )‐ or (R )‐isoleucine were synthesised from the diastereomeric conjugate of (3R , 7R )‐jasmonic acid and (3S , 7S )‐jasmonic acid by sodium borohydride reduction. The resulting diastereomers were characterised by nuclear magnetic resonance spectra. The authentic substances were subjected to liquid chromatography using a reversed‐phase C18 matrix and the chiral stationary phase Chiralpak AS. For all (6RS )‐hydroxy epimeric pairs, a baseline separation could be observed. The elution sequences obtained indicate that the 3,6‐trans oriented epimers with (S )‐isoleucine elute prior to the 3,6‐cis configured individuals independent of the mode of chromatography. In contrast, the elution pattern of the conjugates containing (R )‐isoleucine was reversed on the chiral stationary phase. The epimers with a 3,6‐cis configured hydroxy group and the acid side chain eluted faster than those with the 3,6‐trans configuration. The chromatographic conditions described are suitable for resolving mixtures of isomeric N‐(cucurbinoyl)‐isoleucines in order to assign their stereochemistry and to obtain chiral reference materials on a preparative scale.