The recovery of biophenols from wastewaters of olive oil production

In this article, the phenolic composition of wastewaters prepared from different cultivars of Olea europaea have been described. The main aim is the recovery of these compounds for technological utilization.     

Phenolic components of Olea europaea--isolation of tyrosol derivatives

A new tyrosol derivative, the oleic ester of tyrosol, was isolated from fruits of Olea europaea, Cassanese cultivar. Salidroside, the known glucosyl derivative of thyrosol, was isolated from the Carolea cultivar.     

UV-Vis光谱差减法测定油橄榄叶中橄榄苦苷含量初探

建立了紫外-可见(UV-Vis)分光光谱测定油橄榄叶中橄榄苦苷含量的方法.以芦丁和橄榄苦苷为对照品,分别采用NaNO2-Al(NO3)3-NaOH络合体系和AlCl3络合体系进行显色.以NaNO2-Al(NO3)3-NaOH体系显色后于510 nm处可测得油橄榄叶中总黄酮和橄榄苦苷的吸光度之和;而以AlCl3体系显色后...     

不同石斛枫斗中酚酸类活性成分的比较及杓唇石斛素和石斛酚含量的测定

建立了基于液相色谱-离子阱飞行时间串联质谱(LC-IT-TOF MS/MS)的酚酸类分析方法,对6种共18批次的石斛枫斗药材中的酚类化合物进行了研究,其中18种酚酸类化合物得到初步定性。在共有化合物中选择杓唇石斛素和石斛酚作为质量控制的目标化合物。在此基础上,采用响应面法(RSM)确定了提取溶剂为60%甲醇、提取时间为65 min的超声提取杓唇石斛素和石斛酚的最佳条件,建立了石斛枫斗中杓唇石斛素、石斛酚的高效液相色谱(HPLC)含量测定方法。定量线性优于0.999 8,检出限(LOD)分别为0.18 mg/L和0.09 mg/L,重复性的相对标准偏差(RSD)小于3%,加标回收率均值分别为97.1%和101.4%。这些结果表明本研究建立的方法可用于石斛枫斗药材中酚酸类化学成分研究及质量控制。     

Analysis of phenolic compounds in Epimedium plants using liquid chromatography coupled with electrospray ionization mass spectrometry

A new method based on HPLC coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) has been established for the analysis of phenolic compounds in Epimedium plants. The characteristic fragmentation pathways of 29 phenolic compounds were studied, and 126 compounds were identified or tentatively characterized based on their mass spectra from fourteen samples including the aerial samples and the underground parts of the seven species. Furthermore, the differences in phenolic compound profiles between different Epimedium plants and two parts of the seven species were reported for the first time. Due to their significant chemical differences, these Epimedium species should be used separately in clinical practice.     

Isolation and identification of radical scavengers in olive tree (Olea europaea) wood

Several extracts of Olea europaea wood (Picual olive cultivar) were obtained with solvents of different polarity and their antioxidant activities determined. The active compounds were detected in fractions of an ethyl acetate extract using HPLC with on-line radical scavenging detection. After applying different separation techniques, hydroxytyrosol, tyrosol, cycloolivil, 7-deoxyloganic acid, oleuropein and ligustroside were isolated and characterized. Hydroxytyrosol showed a higher activity than the natural antioxidant rosmarinic acid in scavenging the DPPH model radical. Cycloolivil and oleuropein showed stronger activities than the synthetic antioxidant BHT against the same radical. Ligustroside, tyrosol and 7-deoxyloganic acid showed little activity. The latter compound has not been previously identified in the genus Olea.     

A validated HPLC method for the quantitative analysis of trans-resveratrol and trans-piceid in Hungarian wines

A new, simple procedure for the determination of phenolic components of wines, such as resveratrol and piceid isomers, has been elaborated and validated. A set of 70 red wines and 3 white wines from two wineries (Polgar Winery and Bock Winery, Villany, Hungary) are analyzed by reversed-phase high-performance liquid chromatography. The samples are injected without pretreatment and UV-vis and mass spectrometric (MS) detection has been applied. The detection limit for trans-resveratrol and for trans-piceid is found to be 0.9 and 0.6 pmol for the UV-vis detection method and 0.3 and 0.2 pmol for the MS detection method. Trans-resveratrol and trans-piceid are found in red wines from 0.1 to 14.3 mg/L and from 3.8 to 16.4 mg/L concentrations, respectively.     

A protein extraction method compatible with proteomic analysis for the euhalophyte Salicornia europaea

Protein extraction from plants like the halophyte Salicornia europaea has been problematic using standard protocols due to high concentrations of salt ions in their cells. We have developed an improved method for protein extraction from S. europaea, which allowed us to remove interfering compounds and salt ions by including the chemicals borax, polyvinylpolypyrrolidone, and phenol. The comparative study of this method with several other protocols using NaCl-treated S. europaea shoots demonstrated that this method gave the best distinction of proteins on 2-DE gels. This protocol had a wide range of applications as high yields and good distinction of 1-DE gels for proteins isolated from twelve other plants were rendered. In addition, we reported results of 2-DE using the recalcitrant tissue of the S. europaea roots. We also demonstrated that this protocol is compatible with proteomic analysis as eight specific proteins generated by this method have been identified by MS. In conclusion, our newly developed protein extraction protocol is expected to have excellent applications in proteomic studies of halophytes.     

Screening non-colored phenolics in red wines using liquid chromatography/ultraviolet and mass spectrometry/mass spectrometry libraries

Liquid chromatography/ultraviolet (LC/UV) and mass spectrometry/mass spectrometry (MS/MS) libraries containing 39 phenolic compounds were established by coupling a LC and an ion trap MS with an electrospray ionization (ESI) source, operated in negative ion mode. As a result, the deprotonated [M-H]- molecule was observed for all the analyzed compounds. Using MS/MS hydroxybenzoic acid and hydroxycinnamic acids showed a loss of CO2 and production of a [M-H-44]- fragment and as expected, the UV spectra of these two compounds were affected by their chemical structures. For flavonol and flavonol glycosides, the spectra of their glycosides and aglycones produced deprotonated [M-H]- and [A-H]- species, respectively, and their UV spectra each presented two major absorption peaks. The UV spectra and MS/MS data of flavan-3-ols and stilbenes were also investigated. Using the optimized LC/MS/MS analytical conditions, the phenolic extracts from six representative wine samples were analyzed and 31 phenolic compounds were detected, 26 of which were identified by searching the LC/UV and MS/MS libraries. Finally, the presence of phenolic compounds was confirmed in different wine samples using the LC/UV and LC/MS/MS libraries.     

当归化学成份分析研究——毛细管气相色谱-质谱法鉴定当归根挥发油成份

当归具有活血化瘀、镇静、解痉等作用.分析其化学成份对研究其药理作用具有极重要的意义.自1924年开始,即有这方面的研究报导^[1-11].     

超高效液相色谱-四极杆-飞行时间质谱定性研究茶叶籽中的酚类化合物

采用超高效液相色谱(UHPLC)-四极杆-飞行时间质谱(Q-TOF/MS)技术定性分析茶叶籽中的酚类化合物。茶叶籽样品经乙醇水溶液提取后经反相色谱分离,通过Q-TOF/MS进行化合物的鉴定。基于山茶属及相关植物化学组成的文献,建立了一个含有106种酚类化合物的数据库。对UHPLC-Q-TOF/MS采集得到的一级质谱数据进行数据库检索,然后对检索到的化合物色谱峰进行二级质谱扫描,根据得到的碎片离子推断化合物的结构。初步推断出茶叶籽提取物中的24种酚类化合物,包括13种酚酸类、4种儿茶素类和7种黄酮类化合物,并通过与标准品比对,进一步确证了这些化合物。结果表明UHPLC-Q-TOF/MS技术可以用于对茶叶籽中酚类化合物进行快速、准确、可靠的定性分析,促进新化合物的发现与鉴别。     

Rhamnus pallasii subsp. sintenisii fruit,leaf, bark and root: Phytochemical profiles and biological activities

The genus Rhamnus has received a lot of interest as a source of phenolic chemicals. There have been no reports on the phytochemicals and biological activities of R. pallasii subsp. sintenisii various morphological components (fruit, leaf, bark, and root) in Iran to yet. Two crude ether petroleum (EP) and hydro-methanolic (HM) extracts were obtained from the separate parts. The antioxidant and antibacterial capabilities of the extracts, as well as their phytochemical screening (total phenolic, flavonoid, phenolic acid, and anthocyanin concentrations), were measured. Furthermore, the phytochemical profiles of EP and HM extracts were determined using GC–MS and LC-ESI–MS, respectively. LC-ESI-MS detected 59 chemicals in HM extracts, including flavonoids (62.71 %), phenolic acids (10.16 %), and anthraquinones (16.94 %). Furthermore, the predominant group components in EP extracts examined by GC–MS were fatty acids (58.82%), phenolic compounds (49.28%), and hydrocarbons (35.15 to 59.45 %). In terms of biological testing (DPPH radical scavenging and anti-bacterial activity), all examined extracts, particularly the fruit, had the highest activities in both assays (IC₅₀: 7.52 to 22.39 µg/ml and MIC: 0.39 to 3.12 mg/ml), owing to their high phenolic content. As a result, individual morphological elements of the species might be thought of as natural antioxidant and antibacterial agents.     

Phenolic compounds of mountain tea from the Balkans: LC/DAD/ESI/MSn profile and content

Twenty-one samples of Sideritis species (S. scardica, S. raeseri, S. taurica, S. syriaca and S. perfoliata) from various locations on the Balkan Peninsula were evaluated for their chemical constituents. Chemical analyses were focused on secondary metabolites, particularly phenolic compounds, which have several roles in the plant physiological processes and have demonstrated significant health beneficial effects. The occurrence of hydroxycinnamic acids, phenylethanoid glycosides and flavonoids has been investigated in taxonomically related taxa of the genus Sideritis. A systematic method for phenolic compounds identification was developed using tandem mass spectrometry coupled to high performance liquid chromatography with diode array detection. Scanning for precursor ions of commonly found phenolics in Sideritis species using LC/MS11 with an ion trap instrument permitted the specific determination of hydroxycinnamic acid derivatives, and phenylethanoid and flavonoid glycosides. Further characterization of each phenolic compound was performed using MS/MS product-ion analysis and common-neutral-loss analysis. This on-line technique allowed identification of three hydroxycinnamic acid derivatives, eight phenylethanoid glycosides, and twenty-four flavonoid glycosides. All the taxa analysed produced very similar phenolic patterns characterized by the presence of 5-caffeoylquinic acid, lavandulifolioside, verbascoside, hypolaetin 7-O-[6'-O-acetyl]-allosyl(1-->2)glucoside, apigenin 7-(4"-p-coumaroylglucoside), 4'-O-methylisoscutellarein 7-O-[6'-O-acetyl]-allosyl(1-->2)glucoside, and minor amounts of isoverbascoside, apigenin 7-O-allosyl(1-->2)glucoside, isoscutellarein 7-O-allosyl-(1-->2)-[6"-O-acetyl]-glucoside, hypolaetin 7-O-allosyl-(1-->2)-[6"-O-acetyl]-glucoside and 4'-O-methylhypolaetin 7-O-[6'-O-acetyl]-allosyl-(1-->2)-[6"-O-acetyl]-glucoside. These results show that the investigated species are systematically very closely related. Phenylethanoid glycosides and flavonoid acetylglycosides are dominant and constitute 90% of the total phenolic compounds compared with hydroxycinnamic acid and flavonoid 7-O-glycosides. Principal component analysis (PCA) was performed for the nature and content of the different compounds to be correlated to the particular Sideritis species and also to the locations.     

Structural differentiation of uronosyl substitution patterns in acidic heteroxylans by electrospray tandem mass spectrometry

The structures of two oligomers of acidic xylo-oligosaccharides (XOS) of the same molecular weight (634 Da), Xyl(2)MeGlcAHex and Xyl(2)GlcA(2) were differentiated by electrospray tandem mass spectrometry (ESI-MS/MS). These oligomers were present in a mixture of XOS obtained by acid hydrolysis of heteroxylans extracted from Eucalyptus globulus wood (Xyl(2)MeGlcAHex) and Olea europaea olive fruit (Xyl(2)GlcA(2)). In the ESI-MS spectra of the XOS, ions at m/z 657 and 652 were observed and assigned to [M + Na](+) and [M + NH(4)](+), respectively. The ESI-MS/MS spectrum of [M + Na](+) ion of Xyl(2)MeGlcAHex showed the loss of Hex residue from the reducing end followed by the loss of MeGlcA moiety. Simultaneously, the loss of a Xyl residue from either the reducing or the non-reducing ends was detected. On the other hand, the fragmentation of Xyl(2)GlcA(2) occurs mainly by the loss of one and two GlcA residues or by the loss of the GlcAXyl moiety, due to the glycosidic bond cleavage between the two Xyl residues. Loss of one and two CO(2) molecules was only observed for this oligomer, where the GlcA are in vicinal Xyl residues. The ESI-MS/MS spectra of [M + NH(4)](+) of both oligomers showed the loss of NH(3), resulting in the protonated molecule, where the presence of ions assigned as protonated molecules of aldobiuronic acid residues, [MeGlcA - Xyl + H](+) and [GlcA - Xyl + H](+), are diagnostic ions of the presence of MeGlcA and GlcA moieties in XOS. Since these structures occur in small amounts in complex acidic XOS mixtures and are very difficult, if possible, to isolate, tandem mass spectrometry revealed to be a powerful tool for the characterization of these types of substitution patterns present in heteroxylans.     

Production of triterpene acids by cell suspension cultures of Olea europaea

Olive (Olea europaea) contains large quantity of triterpene acids including oleanolic acid (6) as a major one. Varieties of biological activities exhibited by triterpene acids attracted our attentions, especially from pharmaceutical viewpoints. Cell culture of olive plant was induced and its triterpene constituents were studied. From the cell suspension cultures, six ursane type triterpene acids; ursolic acid (9), pomolic acid (10), rotundic acid (11), tormentic acid (12), 2alpha-hydroxyursolic acid (13) and 19alpha-hydroxyasiatic acid (14), and two oleanane type acids; oleanolic acid and maslinic acid (7), have been isolated. Quantity of ursane type triterpene acids produced by cell cultures was larger than that of oleanane type. Further, a multifunctional oxidosqualene cyclase (OSC) named OEA was cloned by homology based PCRs from the same cultured cells. Major product of OEA is alpha-amyrin (ursane skeleton), showing good accordance to higher content of ursane-type triterpene acids in the cultured cells, and strongly suggesting OEA to be a major contributor OSC for their production.     

β-D-吡喃型葡萄糖酚苷类天然类似物的合成及抗HCV病毒活性

以α-溴代四乙酰基葡萄糖(或β-D-葡萄糖五乙酸酯)和酚为原料,经2步反应合成了15个新的β-D-吡喃型葡萄糖酚苷类天然产物类似物,采用质谱(MS)及核磁共振(NMR)等手段对其结构进行了表征.生物活性测定结果表明,化合物8a和10a对丙型肝炎病毒(HCV)有一定的抑制作用,它们的EC50值分别为21和9.4μmol/L,CC50值分别为150.4和145.9μmol/L.     

Identification and quantitation of phenolic compounds in faecal matrix by capillary gas chromatography and nano-electrospray mass spectrometry

Very few relevant methods have been described for the detection and quantitation of phenolic compounds in faecal matrix. Extraction with conventional organic solvents such as chloroform/methanol (2:1, Folch reagent), methanol and ethanol (72%) showed high extraction efficiency for lipids and also gave good recovery of the major phenolic compounds present in the matrix. However, in comparison with a newly developed phosphate buffer method, the yield of minor phenolics was negligible when detected by these conventional methods. Conventional methods also lead to contamination of the ion source of the mass spectrometer and rapid deterioration of column performance mostly due to the high concentration of lipids. However, if the faecal matrix is initially extracted with phosphate buffer, and the extract acidified and re-extracted with diethyl ether, the range and yield of phenolic compounds are enhanced and the problem of lipid contamination is substantially alleviated. Following pilot studies and optimisation of the procedure, individual phenolic compounds (n = 29) were identified by nano-electrospray ionisation mass spectrometry (nano-ESI-MS), nano-ESI-tandem mass spectrometry (MS/MS) and gas chromatography/mass spectrometry (GC/EI-MS) and quantitated (n = 27) by GC/MS in subsets (n = 5) of faecal samples, collected during the European Agency for Cancer Prevention calcium/fibre intervention study from four European countries (Italy, Germany, Spain and Denmark). A range of phenolic compounds (mainly acids) was detected, dominated by phenylacetic, benzoic, phenylpropionic and m-hydroxyphenylpropionic acids, representing on average 9.91 (93%), 8.25 (92%), 9.45 (95%) and 11.05 (98%) mM in the Italian, German, Spanish and Danish samples, respectively. The new method should enable large epidemiologic, case-control and intervention studies on the relevance of phenolic antioxidants in the aetiology of colorectal cancer to be conducted in the future.     

Establishment of a UPLC-PDA/ESI-Q-TOF/MS-Based Approach for the Simultaneous Analysis of Multiple Phenolic Compounds in Amaranth (A. cruentus and A. tricolor)

We used ultraperformance liquid chromatography coupled with a photodiode-array detector and electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-PDA/ESI-Q-TOF/MS) to rapidly and accurately quantify 17 phenolic compounds. Then, we applied this method to the seed and leaf extracts of two Amaranthus species to identify and quantify phenolic compounds other than the 17 compounds mentioned above. Compounds were eluted within 30 min on a C18 column using a mobile phase (water and acetonitrile) containing 0.1% formic acid, and the specific wavelength and ion information of the compounds obtained by PDA and ESI-Q-TOF/MS were confirmed. The proposed method showed good linearity (r² > 0.990). Limits of detection and quantification were less than 0.1 and 0.1 μg/mL, respectively. Intra- and interday precision were less than 2.4% and 1.8%, respectively. Analysis of amaranth seed and leaf extracts using the established method showed that the seeds contained high amounts of 2,4-dihydroxybenzoic acid and kaempferol, and leaves contained diverse phenolic compounds. In addition, six tentatively new phenolic compounds were identified. Moreover, seeds potentially contained 2,3-dihydroxybenzaldehyde, a beneficial bioactive compound. Thus, our method was an efficient approach for the qualitative and quantitative analysis of phenolic compounds, and could be used to investigate phenolic compounds in plants.     

A general analytical strategy for the characterization of phenolic compounds in fruit juices by high-performance liquid chromatography with diode array detection coupled to electrospray ionization and triple quadrupole mass spectrometry

In the present study, a methodology based on liquid chromatography with diode array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer for the simultaneous identification of phenolic compounds in fruit juices has been developed. 72 available phenolic compound standards from diverse families present in fruits have been studied in order to analyze their fragmentation pattern. As a result, a general strategy for the characterization of unknown phenolic compounds in fruit juices was designed: (i) taking into account its UV–visible spectrum and elution order, assign the unknown polyphenol to a polyphenol class, (ii) identify the quasi-molecular ion using positive and negative MS spectra, being supported by adducts generated with solvent or sodium and molecular complexes, (iii) determinate the pattern of glycosylation in positive mode using ESI(+)-CID MS/MS product ion scan experiments, selecting the quasi-molecular ion as precursor ion, and finally, (iv) study the identity of the aglycone through ESI(+)-CID MS/MS product ion spectra from the protonated aglycone, [Y₀]⁺. This strategy was successfully employed for the characterization of known and unknown phenolic compounds in juices from 17 different fruits.     

Investigating tree bark as an air-pollution biomonitor by means of neutron activation analysis

The olive tree (Olea europaea) is an icon of southern Europe and awidespread evergreen in mainland Portugal. First results of a continuing studyon the ability of olive-tree bark to act as an air-pollution biomonitor arepresented and discussed here. Other than lower signals and an anticipatedsystemic control over some elements, there seems to be no a priori reasonfor ruling out the possibility of using bark in atmospheric trace-elementsurveys. In fact, nonparametric statistics show that, despite their relativemagnitude, the variation patterns of bark and lichen concentrations significantlyfollow one another all across the study area.