Improved sample preparation for CE-LIF analysis of plant N-glycans

In view of glycomics studies in plants, it is important to have sensitive tools that allow one to analyze and characterize the N-glycans present on plant proteins in different species. Earlier methods combined plant-based sample preparations with CE-LIF N-glycan analysis but suffered from background contaminations, often resulting in non-reproducible results. This publication describes a reproducible and sensitive protocol for the preparation and analysis of plant N-glycans, based on a combination of the 'in-gel release method' and N-glycan analysis on a multicapillary DNA sequencer. Our protocol makes it possible to analyze plant N-glycans starting from low amounts of plant material with highly reproducible results. The developed protocol was validated for different plant species and plant cells.     

Pressurized capillary electrochromatography in a screening for possible antioxidant molecules in Mallotus fingerprints: challenges, potentials and prospects

Because of its eminent high resolution potential and minimal solvent consumption, pressurized capillary electrochromatography (pCEC) may offer an interesting alternative to HPLC for screening applications that need to resolve complex samples. In this paper, its potential was assessed in a screening of plant extracts from Mallotus species to indicate compounds with possible antioxidant activities by means of a PLS model built from their pCEC fingerprints. The main aim of this research was to find out whether pCEC can have an added value for this application. To get a complete overview of the techniques potential for this application, it was also assessed whether the technique can meet the requirements in terms of precision, sensitivity and column robustness. Encountered benefits and downsides were reported. Fingerprints with satisfactory sensitivity and precision could be obtained by concentrating the sample 5-fold and using optimized rinsing procedures, respectively. From the generated pCEC fingerprints of 39 Mallotus samples and their respective DPPH radical scavenging activity test results, a three-component PLS model was being built. The model proved good predictive abilities and easily allowed the indication of possible antioxidant compounds in the fingerprints. Despite its much higher peak capacity, the performance of pCEC to fingerprint the majority of the Mallotus extracts did not surpass that of a custom HPLC method. This was also reflected in its comparable power to indicate possible antioxidant compounds in the fingerprints after modeling. Because of its low detection sensitivity and modest column robustness, the benefit of the lower solvent consumption was partly paid-off by the current need for more system maintenance, also limiting the sample throughput. For the considered screening application, pCEC may suit as a viable but no preferred alternative technique.     

Reduction of 4-(haloalkyl)azetidin-2-ones with LiAlH4 as a powerful method for the synthesis of stereodefined aziridines and azetidines

[reaction: see text] A new synthesis of stereodefined aziridines and azetidines, starting from 4-(1- or 2-haloalkyl)azetidin-2-ones, is described. Treatment of the latter compounds with LiAlH(4) gave 1,2-fission of the beta-lactam, followed by an intramolecular nucleophilic substitution of the halogen, giving rise to the formation of 2-(1-alkoxy-2-hydroxyethyl)aziridines in the case of 4-(1-haloalkyl)azetidin-2-ones and of 2-(1-alkoxy-2-hydroxyethyl)azetidines in the case of 4-(2-haloalkyl)azetidin-2-ones. The resulting 2-(1-alkoxy-2-hydroxyethyl)aziridines were transformed into the corresponding trans-3,4-substituted oxolanes via an intramolecular nucleophilic ring opening, triggered by AlCl(3).