Opuntisines, 14-membered cyclopeptide alkaloids from fruits of Opuntia stricta var. dillenii isolated by high-performance countercurrent chromatography.

Extracts of Opuntia stricta var. dillenii fruits were fractionated by semi-preparative high-performance countercurrent chromatography (HPCCC) to study the secondary metabolite formation, whereby HPCCC showed a superior separation capacity to fractionate minor metabolites compared to HPLC. A family of new peptides was detected in semi-polar fractions when monitoring the HPCCC separation by off-line injections of fractions to ESI-MS/MS. Planar structures of the major compounds, two 14-ring-membered cyclopeptide alkaloids, which were named opuntisines A and B, were elucidated by 1D- and 2D-NMR spectroscopy and HR-ESI-MS/MS spectrometry, while a combination of chemical derivatisation and degradation revealed the stereo-configurations. Specifically, the methods of Marfey and Mosher indicated l-Glu, l-Ile, l-Phe and 1S-configurations, respectively; ROESY correlations revealed 8S, 9S. The novel opuntisine A showed moderate activity against the Gram-negative bacterium Escherichia coli, but no further antibacterial, antifungal nor cytotoxic effects. This bioactive natural product class is reported for the first time in the plant family Cactaceae.

Profiling of polar metabolites in fruits of Opuntia stricta var. dillenii by ion-pair high-performance countercurrent chromatography and off-line electrospray mass-spectrometry injection.

Electrospray mass spectrometry-profiling guided the metabolome investigation of a C18 reversed phase adsorbate of Opuntia stricta var. dillenii fruits following analytical, and semi-preparative high-performance countercurrent chromatography (HPCCC) fractionation, and visualization of molecular weight elution profiles based on selected single ion-traces. Experimental partition-ratio values KD, and peak widths for detected metabolites were determined. Structural characterization of metabolites, and co-elution effects were monitored in the scan range m/z 150-2200. The polar ion-pair activated solvent system tert.-butylmethylether - n-butanol - acetonitrile - water (0.7% trifluoroacetic acid) [2:2:1:5]) was used for partition-ratio KD improvement of ionic betalains. HPCCC operations were in the head-to-tail mode using the elution-extrusion approach. Selected ESI-MS ion traces visualized the elution of fourty-three metabolites, whereas twenty-one were identified as known betalain pigments, and their classic degradation products, and chlorinated betacyanin artefacts. Potentially, novel fruit metabolites of Opuntia were recognized by unknown molecular weights, and MS/MS fragmentations. Off-line ESI-MS fraction monitoring determined peak elution windows, and resulted in KD-based chromatographic scales. Detectable metabolites were compared by separation- α, and resolution-factors RS revealing a better performance of the analytical HPCCC experiment. Experimental metabolite KD-values from analytical and semi-preparative HPCCC runs were widely consistent, and confirmed the reproducibility of the technique based on the used sample material.

Technological characteristics and selected bioactive compounds of Opuntia dillenii cactus fruit juice following the impact of pulsed electric field pre-treatment.

Selected technological characteristics and bioactive compounds of juice pressed directly from the mash of whole Opuntia dillenii cactus fruits have been investigated. The impact of pulsed electric fields (PEF) for a non-thermal disintegration on the important juice characteristics has been evaluated in comparison to microwave heating and use of pectinases. Results showed that the cactus juice exhibited desirable technological characteristics. Besides, it also contained a high amount of phenolic compounds being the major contributors to the overall antioxidant activity of juice. HPLC-DAD/ESI-MS(n) measurements in the fruits' peel and pulp showed that isorhamnetin 3-O-rutinoside was determined as the single flavonol found only in the fruit's peel. Treating fruit mash with a moderate electric field strength increased juice yield and improved juice characteristics. Promisingly, the highest release of isorhamnetin 3-O-rutinoside from fruit's peel into juice was maximally achieved by PEF.

Influence of cultivar and origin on the flavonol profile of fruits and cladodes from cactus Opuntia ficus-indica.

Flavonols are hypothesized to be the most important polyphenolic antioxidants. This present study aimed at investigating flavonols in cactus Opuntia ficus-indica fruits from two Egyptian cultivars in comparison to three common Sicilian cultivars, and two further cultivars from South Africa. Besides, cactus O. ficus-indica cladodes from Egyptian cultivars have been investigated as well. HPLC-DAD analyses showed that cactus O. ficus-indica fruits and cladodes are mainly characterized by isorhamnetin glycosides. These flavonols were found only in the fruit's peel and the cladodes, but not in the pulp. However, all analyzed cultivars exhibited the same flavonol profile which might therefore serve as a chemical fingerprint with regard to genuineness of cactus O. ficus-indica fruits and cladodes or even food products containing whole cactus fruits as ingredients. The findings obtained have been confirmed both by enzymatic hydrolysis of flavonol glycosides following analysis of the aglycons and by HPLC-ESI-MSn. The concentration of total flavonols ranged from 2.2 to 4.1 and 6.3 to 7.6mg/g (dw) in the fruit peels and the cladodes, respectively. Further, fruit peels and cladodes exhibited high total phenolic contents and antioxidant activities compared to those of the fruit pulps. These investigations valorize cactus fruit's peel and cladode in comparison to fruit's pulp, and may further provide additional data for using flavonols in chemotaxonomic studies of cactus Opuntia spp. and authenticity of cactus O. ficus-indica products.