•Techniques of different sensitivities were combined to identify/quantify bioactive compounds.•Identification/quantification of bioactive structural isomers in salvia, oregano, thymus and apple.•Good ...correlation of quantitative results obtained by HPTLC-Vis and HPTLC-1H qNMR-PULCON.•Comparison of HPTLC-1H qNMR-PULCON versus 1H deconvoluted versus 1H–13C HSQC.•Low solvent volume consumed compared to bioactivity-guided fractionation.
Differing sensitivity is the main obstacle for a direct combination of HPTLC with NMR spectroscopy. A sufficient amount of the isolated compound zone must be provided by HPTLC for subsequent offline NMR detection (HPTLC//NMR). To fill the gap, a straightforward procedure was developed using the same analytical HPTLC system for both bioprofiling and isolation of bioactive zones from multicomponent mixtures. The HPTLC-effect-directed analysis (EDA) revealed several bioactive compounds in five botanical extracts, i.e. Salvia officinalis, Thymus vulgaris and Origanum vulgare, all Lamiaceae, and peels of red and green apples (Jonagored and Granny Smith, respectively), both Rosaceae. A tricky case study was designed to show how to deal with potentially coeluting bioactive structural isomers, e.g., ursolic (UA), oleanolic (OA) and betulinic acids (all C30H48O3), which are most difficult to identify and assign. A multipotent bioactive HPTLC zone showed the same hRF value and mass signal in HPTLCHRMS, though containing the coeluting structural isomers UA and OA. After zone isolation from the HPTLC plate, first the 1H NMR spectrum allowed to distinguish distinct allylic H-18 protons, i.e. 2.20 ppm for UA and 2.85 ppm for OA, and at the same time, to quantify the two isomers by using the PUlse Length-based CONcentration methodology (HPTLC//1H qNMR-PULCON). In case of a partial overlap of the diagnostic signal with that of the matrix, results were corroborated with those obtained by using the 1H deconvoluted or 2D 1H–13C Heteronuclear Single Quantum Coherence spectra. The comparison of the quantitative results showed a good correlation (R2 = 0.9718) between the two orthogonal methods HPTLC-Vis and HPTLC//1H qNMR-PULCON. A sufficient zone isolation from the HPTLC plate (mean isolation rate of 82%) for both UA and OA (0.27 - 4.67 mM) was achieved for HPTLC//qNMR, comparing the isolated bioactive compound zone with the respective zone in the botanical extract via HPTLC-Vis densitometry. The HPTLC-EDA-Vis//1H qNMR-PULCON procedure for bioprofiling and quantification/identification/confirmation of bioactive compounds in botanical extracts is considered as straightforward, eco-friendly (only 16 mL solvent required), simple (NMR calibration used over weeks) and reliable new alternative to the status quo of bioactivity-guided fractionation.
Essential oils from oregano (Origanum vulgare L.), mint (Mentha piperita L.), fennel (Foeniculum vulgare Mill.), and pine (Abies alba Mill.) needles and cones, and their active substances thymol, ...carvacrol, menthol, and anisaldehyde were tested for antifungal activity against Penicillium verrucosum. The lowest minimal inhibitory concentrations (MICs) were achieved for essential oil of oregano, followed by carvacrol, thymol, and menthol. These antifungal components were further investigated, as the main aim of our study was to elucidate the effect of natural antifungals on ochratoxin A production. During 21 days of exposure, the growth of P. verrucosum, and subsequently the production of ochratoxin A, was fully inhibited by thymol at ½ MIC (0.0625 mg mL
), but menthol at ¼ and ½ MIC (0.1875 and 3750 mg mL
) showed no growth inhibition. After 21 days of incubation, the greatest inhibitory effect on ochratoxin production (inhibition was 96.9 %) was also achieved with thymol at ¼ MIC (0.0313 mg mL
). Essential oil of oregano (¼ MIC, 0.2930 μL mL
) and carvacrol (½ MIC, 0.1953 μL mL
) stimulate production of ochratoxin A at 13.9 % to 28.8 %, respectively. The observed antifungal effects depended on the agent, the concentration used, and the time of interaction between the agent and P. verrucosum. Our results indicate the possibility of using oregano essential oil as a substitute for artificial preservatives in certain foods, but further research is needed.
Eterična olja origana (Origanum vulgare L.), mete (Mentha piperita L.), janeža (Foeniculum vulgare Mill.), in borovih (Abies alba Mill.) iglic in storžev, ter njihove sestavine timol, karvakrol, mentol in anisaldehid smo testirali kot snovi z protiglivno aktivnostjo proti plesnim vrste Penicillium verrucosum. Najnižje minimalne inhibitorne koncentracije (MICs) smo določili eteričnemu olju origana, ter karvakrolu, timolu, in mentolu. Rast plesni vrste P. verrucosum je bila popolnoma inhibirana po 21 dneh inkubacije v prisotnosti ½ MIC timola (0,0625 mg mL-1), medtem ko so eterično olje origana (½ MIC, ¼ MIC), timol (¼ MIC) in karvakrol (½ MIC, ¼ MIC) inhibirali rast plesni od 8,3 % do 70.8 %. Dodatek mentola v koncentracijah ¼ MIC in ½ MIC (0.1875 mg mL
in 0.3750 mg mL
) pa ni imel inhibitornega vpliva na rast plesni vrste P. verrucosum. Popolno inhibicijo tvorbe ohratoksina A smo določili le v primeru, ko je bila tudi rast teh plesni popolnoma inhibirana (pri ½ MIC timola). Največjo inhibicijo tvorbe ohratoksina A (96,9 %) po 21 dnevih inkubacije smo določili pri ¼ MIC timola (0.0313 mg mL
). Pospešeno tvorbo ohratoksina A (od 13,9 % do 28,8 %) pa smo določili pri ¼ MIC eteričnega olja origana (0.2930 mg mL
) in ½ MIC karvakrola (0.1953 mg mL
). Protiglivni vplivi so bili odvisni od testirane snovi, njene koncentracije ter časa interakcij med protiglivno snovjo in plesnimi vrste P. verrucosum. Rezultati nakazujejo možnosti uporabe eteričnega olja origana v živilih namesto kemijskih konzervansov. Ker pa so živila kompleksna okolja v katerih je potrebno upoštevati več dejavnikov, so potrebne nadaljnje raziskave.
This study aimed to determine the effects of different inclusion levels of oregano into the diet on daily feed consumption, feed conversion ratio, egg production, hatchability, egg palatability, egg ...weight, external and internal egg quality, live weight increase, serum cholesterol and yolk cholesterol level in laying quails.
A total of 216 laying quails at 42 days of age were divided into 36 cages, 6 birds in each (4 females and 2 males), 6 cages per thesis. The 1
st
group was left as control (OR0) without any supplementation of oregano. The other five experimental groups were OR10, OR20, OR30, OR40, and OR50 which were fed with the diets supplemented with 10, 20, 30, 40, and 50 g/kg of oregano, respectively. The experiment lasted for 63 days until the quails reached 105 days of age.
As a result of this study, 50 g/kg oregano supplementation appeared to be the best as respects taste (P<0.002). The inclusion level of 20 g/kg increased fertility (P<0.02) however, above that level fertility was reduced. This peculiarity of oregano may be taken into account by the poultry breeder companies. For the other segments of the poultry industry, the usage of oregano up to 50 g/kg seemed to be appropriate.
Oregano essential oil with and without the addition of EDTA was tested against Escherichia coli. The amount of essential oil, the addition of EDTA and the incubation temperature of the medium ...influenced the maximum growth rate (micron sub(max)) and the lag phase of Escherichia coli O157:H7. In particular, the presence of oregano essential oil at the time of inoculation caused an increase in the lag phase, a decrease in the maximum growth rate and a reduced final population size. The inhibitory effect on E. coli O157:H7 is probably associated only with the oregano essential oil, since there was no antibacterial action in broth samples inoculated with E. coli O157:H7 and supplemented alone with EDTA. However, the combination of the two was found to increase the inhibition dramatically. The minimum bactericidal concentration of oregano essential oil on cells of E. coli O157:H7 with and without EDTA affected the dense cellular materials and coagulated the cytoplasmic constituents, while the membrane structure tended to be irregular and rough
E' stato saggiato su Escherichia coli l'effetto inibitorio dell'olio essenziale di origano e dell'EDTA. La quantita' di olio essenziale, l'aggiunta di EDTA e la temperatura di incubazione del mezzo hanno influenzato la velocita' massima di crescita (micron inf(max)) e la fase di latenza di Escherichia coli O157:H7. In particolare, la presenza di olio essenziale di origano al momento dell'inoculo ha causato un incremento della fase di latenza e una diminuzione della velocita' massima di crescita ed una riduzione della dimensione finale della popolazione microbica. L'effetto inibitorio su E. coli O157:H7 e' probabilmente associato al solo olio essenziale di origano, poiche' non si e' evidenziata alcuna attivita' antibatterica in campioni inoculati con E. coli O157:H7 additivati con EDTA. D'altronde, la combinazione dei due principi attivi ha mostrato un aumento spiccato dell'inibizione. La concentrazione battericida minima dell'olio essenziale di origano su E. coliO157:H7 (con o senza EDTA) ha influenzato il materiale cellulare denso e ha coagulato i costituenti citoplasmatici, mentre la struttura della membrana e' divenuta irregolare e rugosa
Antioxidant activity of oregano (Origanum vulgare L.) leaves Bendini, A; Gallina Toschi, T; Lercker, G. (Bologna Univ. (Italy). Dipartimento di Scienze degli Alimenti)
Italian journal of food science,
(2002), 2002, Volume:
14, Issue:
1
Journal Article
Peer reviewed
Oregano leaves (Origanum vulgare L.) were extracted with ethanol, diethyl ether, n-hexane:2-propanol 4:1 (v/v) and n-pentane. The highest yield was obtained with ethanol and the ethanolic extract ...showed the strongest inhibitory effect on lipid oxidation when added (amounts from 0.02 to 5% w/w) to a model lipid system (made up of refined bleached peanut oil), subjected to forced dynamic oxidation, using the Oxidative Stability Instrument (OSI). The hydrolyzed ethanolic extract showed a more powerful antioxidant activity than the untreated extract. Total polyphenols in this extract, expressed as gallic acid, were quantified by spectrophotometric analysis at 280 nm. The ethanolic extract was also analyzed by TLC (thin layer chromatography) and showed the presence of flavones, flavanones and dihydroflavonols, whether glycosilated or not
Le foglie di origano (Origanum vulgare L.) sono state estratte con etanolo, etere dietilico, n-esano:isopropanolo 4:1 (v/v) e n-pentano. La resa piu' elevata e' stata ottenuta utilizzando l'etanolo e l'estratto etanolico ha mostrato anche il piu' forte effetto inibitorio nei confronti dell'ossidazione lipidica, quando addizionato (in quantita' variabili dallo 0,02 al 5% p/p) ad un sistema lipidico modello (costituito da olio di arachide raffinato e decolorato), sottoposto ad ossidazione dinamica forzata, mediante l'Oxidative Stability Instrument (OSI). L'estratto etanolico idrolizzato ha mostrato una maggiore attivita' antiossidante rispetto all'estratto non trattato. I polifenoli totali di questo estratto, espressi come acido gallico, sono stati quantificati attraverso analisi spettrofotometrica, a 280 nm. L'estratto etanolico e' stato anche analizzato in TLC (cromatografia su strato sottile) e ha mostrato la presenza di flavoni, flavanoni e diidroflavonoli, sia liberi, sia glicosilati
Raw and blanched oregano (Origanum vulgare L. ssp. prismaticum Gaudin) were air-dried at 35 deg C and 50 deg C in two drier pilot plants; raw oregano was also dried at room temperature in a shady ...place. Oregano leaves were analysed for volatile phenolic compounds by GC/MS-SIM, for chlorophyll by HPLC and for colour by reflected-light colorimetry. The highest contents of thymol and carvacrol were found in unblanched oregano dried at 35 deg C, followed by oregano dried at room temperature and by both samples dried at 35 deg C and 50 deg C after blanching. The highest chlorophyll content was identified in oregano dried at room temperature, followed by unblanched oregano dried at 50 deg C and blanched oregano dried at 35 deg C. All unblanched, air-dried oregano leaves had colour attributes similar to the raw ones, while blanched-dried leaves were greener. The least damaging treatments were air-drying at 35 and 50 deg C without blanching
Origano (Origanum vulgare L. ssp. prismaticum Gaudin) fresco o scottato è stato essiccato sia in essiccatore a circolazione di aria forzata a 50 gradi C, sia in essiccatore ad aria deumidificata riciclata a 35 gradi C. Una parte di origano fresco è stata inoltre essiccata naturalmente all'aria a temperatura ambiente. Nelle foglie di origano fresco ed essiccato sono stati determinati i componenti fenolici volatili per via GC/MS-SIM, le clorofille mediante HPLC e il colore con colorimetro a riflettanza. L'origano essiccato a 35 gradi C senza scottatura presenta il maggior contenuto di carvacrolo e timolo, seguito dall'origano essiccato all'aria a temperatura ambiente e dai campioni essiccati dopo scottatura. La massima ritenzione in clorofilla si ha nell'origano essiccato all'aria, seguito dagli essiccati a 35 e 50 gradi C, con e senza scottatura rispettivamente. Tutti gli essiccati non scottati hanno i parametri di colore più vicini a quelli dell'origano fresco; negli essiccati dopo scottatura si ha uno spostamento della tinta verso il verde scuro. I trattamenti meno lesivi sono gli essiccamenti a 35 e a 50 gradi C senza scottatura
The effect of the industrial treatment with microwaves on the antioxidant activity of herbs and spices was evaluated. Samples of black pepper (Piper nigrum L.), oregano (Origanum vulgare L.), basil ...(Ocimum basilicum L.) and sage (Salvia officinalis L.), obtained before and after the microwave treatment, were extracted with an acetone-water mixture (70:30). The extracts were analyzed for their phenolic content. The antioxidant activity was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) and deoxyribose methods and the prooxidant activity was determined by evaluating the reducing power and the iron chelating capacity. The treatment did not modify the antioxidant properties of these herbs and spices, as the majority of the differences between treated and non treated samples were not statistically significant
E' stata determinata l'attività antiossidante di estratti di alcune erbe aromatiche e spezie per valutare l'effetto di un trattamento di sanificazione a microonde a cui vengono sottoposte nel corso della lavorazione industriale. Sono stati analizzati campioni di pepe nero (Piper nigrum L.), origano (Origanum vulgare L.), basilico (Ocimum basilicum L.) e salvia (Salvia officinalis L.), prelevati prima e dopo il trattamento di sanificazione. L'estratto in acetone/acqua 70 ottenuto da ciascun campione è stato analizzato per valutare innanzitutto il contenuto di fenoli totali mediante il noto metodo di Folin-Ciocalteau; sugli stessi estratti sono state quindi valutate l'attività antiossidante mediante i metodi del DPPH (2,2-difenil-l-picrilidrazile) e del desossiribosio e la capacità proossidante mediante la valutazione delle capacità riducente e chelante. Il trattamento a microonde sembra solo marginalmente influenzare l'attività antiossidante; infatti la maggior parte delle differenze riscontrate sono risultate statisticamente non significative
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