On-line identification of sugarcane (Saccharum officinarum L.) methoxyflavones by liquid chromatography-UV detection using post-column derivatization and liquid chromatography-mass spectrometry

Sugarcane (Saccharum officinarum L., Gramineae) bagasse and leaves were investigated for their flavonoid content and transgenic sugarcane ("Bowman-Birk" and "Kunitz") was compared with non-modified ("control") plants. Analyses were carried out by high-performance liquid chromatography coupled to diode array UV detection (LC/UV), also using post-column addition of shift reagents, and tandem MS (atmospheric pressure chemical ionization-MS/MS and collision-induced dissociation-MS). On-line UV and MS data demonstrated the presence of methoxyflavone glycosides and aglycones in a total of seven compounds. Three naturally occurring flavones glycosides and two unusual erythro- and threo-diastereoisomeric flavolignan 7-O-glucosides were identified together with their aglycones.     

Comparative study of the flavonoids of some Verbena species cultivated in Egypt by using high-performance liquid chromatography coupled with ultraviolet spectroscopy and atmospheric pressure chemical ionization mass spectrometry

Verbena rigida L., Verbena tenera Spreng. and Verbena venosa L. were investigated for their flavonoid content. Analysis was carried out by high-performance liquid chromatography coupled to diode array UV detection (LC–UV), using different techniques, also using post-column addition of shift reagents, afforded precise structural information about the position of the free hydroxyl groups in the flavonoid nucleus. LC–MS using atmospheric pressure chemical ionization (APCI) in the positive mode provided the molecular weight, the number of hydroxyl groups, the number of sugars and an idea about the substitution pattern of the flavonoid. On-line UV and MS data demonstrated the presence of orientin, vitexin, isovitexin, luteolin, luteolin 7-O-glucoside, apigenin 7-O-glucoside in addition to luteolin, chryseriol and apigenin aglycones in the three Verbena species with different concentrations. Quantitative determination of flavonoid content revealed the presence of 69.84 mg/g dry sample, 88.26 mg/g dry sample and 85.82 mg/g dry sample total flavonoid compounds in V. rigida L., V. tenera Spreng. and V. venosa L., respectively. The method developed for identification is useful for further chromatographic fingerprinting of plant flavonoids.     

Direct identification of phenolic constituents in Boldo Folium (Peumus boldus Mol.) infusions by high-performance liquid chromatography with diode array detection and electrospray ionization tandem mass spectrometry

A very simple and direct method was developed for the qualitative analysis of polyphenols in boldo (Peumus boldus Mol., Monimiaceae) leaves infusions by high-performance liquid chromatography with diode array detection (HPLC-DAD) and electrospray ionization tandem mass spectrometry (HPLC-MSⁿ). The phenolic constituents identified in infusions of the crude drug Boldo Folium were mainly proanthocyanidins and flavonol glycosides. In the infusions, 41 compounds were detected in male and 43 compounds in female leaf samples, respectively. Nine quercetin glycosides, eight kaempferol derivatives, nine isorhamnetin glycosides, three phenolic acids, one caffeoylquinic acid glycoside and twenty one proanthocyanidins were identified by HPLC-DAD and ESI-MS for the first time in the crude drug. Isorhamnetin glucosyl-di-rhamnoside was the most abundant flavonol glycoside in the male boldo sample, whereas isorhamnetin di-glucosyl-di-rhamnoside was the main phenolic compound in female boldo leaves infusion. The results suggest that the medicinal properties reported for this popular infusion should be attributed not only to the presence of catechin and boldine but also to several phenolic compounds with known antioxidant activity. The HPLC fingerprint obtained can be useful in the authentication of the crude drug Boldo Folium as well as for qualitative analysis and differentiation of plant populations in the tree distribution range.     

Characterisation of flavonols in broccoli (Brassica oleracea L. var. italica) by liquid chromatography-uV diode-array detection-electrospray ionisation mass spectrometry

The flavonoid composition of broccoli inflorescences has been studied by LC/UV-DAD/ESI-MSn. A large number of hydroxycinnamic acid esters of kaempferol and quercetin glucosides has been characterised. The structures of the flavonoid glycosides were analysed after alkaline hydrolysis, and were identified as 3-sophoroside/sophorotrioside-7-glucoside/sophoroside of kaempferol, quercetin and isorhamnetin (this last found in trace amount). These complex quercetin and isorhamnetin glucosides have not been previously characterised in nature. In addition, several less complex glucosides based on the same aglycones have been identified. The effect of sugar substitution and acylation on chromatographic mobility and ESI ionisation and fragmentation are discussed.     

Application of liquid chromatography with mass spectrometry combined with photodiode array detection and tandem mass spectrometry for monitoring pesticides in surface waters

Liquid chromatography with photodiode array detection (LC-DAD) and liquid chromatography with mass spectrometry (LC-MS) are two techniques that have been widely used in monitoring pesticides and their degradation products in the environment. However, the application of liquid chromatography with tandem mass spectrometry (LC-MS-MS) for such purposes, once considered too costly, is now gaining considerable ground. In this study, we compare these methods for the multi-residue analysis of pesticides in surface waters collected from the central and southeastern regions of France, and from the St. Lawrence River in Canada. Forty-eight pesticides belonging to eight different classes (triazine, amide, phenylurea, triazole, triazinone, benzimidazole, morpholine, phenoxyalkanoic), along with some of their degradation products, were monitored on a regular basis in the surface waters. For LC-MS, we used the electrospray ionization (ESI) interface in the negative ionization mode on acidic pesticides (phenoxyalkanoic, sulfonylurea), and the atmospheric pressure chemical ionization (APCI) interface in the positive ionization mode on the remaining chemicals. Different extraction techniques were employed, including liquid-liquid extraction with dichloromethane, and solid-phase extraction using C18-bonded silica and graphitized carbon black cartridges. Eleven of the target chemicals (desethylatrazine, desisopropylatrazine, atrazine, simazine, terbuthylazine, metolachlor, carbendazime, bentazone, penconazole, diuron and isoproturon) were detected by LC-MS at concentrations ranging from 20 to 900 ng/l in the surface waters from France, and six pesticides (atrazine, desethylatrazine, desisopropylatrazine, cyanazine, simazine and metolachlor) were detected by LC-MS and LC-MS-MS at concentrations ranging from 3 to 52 ng/l in the samples drawn from the St. Lawrence River. There was good correlation between the LC-DAD and LC-MS techniques for 60 samples. The slope of the curves expressing the relationship between the results obtained with LC-DAD versus those obtained by LC-MS was near 1, with a correlation coefficient (r) of over 0.93. The identification potential of the LC-MS technique, however, was greater than that of the LC-DAD; its mass spectra, mainly reflecting the pseudomolecular ion resulting from a protonation or a deprotonation of the molecule, was rich in information. The LC-MS-MS technique with ion trap detectors, tested against the LC-MS on 10 surface water samples, gave results that correlated well with the LC-MS results, albeit generating mass spectra that yielded far more information about the structure of unknown substances. The sensitivity of the LC-MS-MS was equivalent to the selected ion monitoring (SIM) acquisition mode in LC-MS. The detection limits of the target pesticides ranged from 20 to 100 ng/l for the LC-MS technique (under full scan acquisition), and from 2 to 6 ng/l for LC-MS-MS. These limits were improved by a factor of almost 10 by increasing the sample volume to 10 l.     

Identification of complex,naturally occurring flavonoid glycosides in kale (Brassica oleracea var. sabellica) by high‐performance liquid chromatography diode‐array detection/electrospray ionization multi‐stage mass spectrometry

Kale is a member of the Brassicaceae family and has a complex profile of flavonoid glycosides. Therefore, kale is a suitable matrix to discuss in a comprehensive study the different fragmentation patterns of flavonoid glycosides. The wide variety of glycosylation and acylation patterns determines the health‐promoting effects of these glycosides. The aim of this study is to investigate the naturally occurring flavonoids in kale. A total of 71 flavonoid glycosides of quercetin, kaempferol and isorhamnetin were identified using a high‐performance liquid chromatography diode‐array detection/electrospray ionization multi‐stage mass spectrometry (HPLC‐DAD/ESI‐MSⁿ) method. Of these 71 flavonol glycosides, 27 were non‐acylated, 30 were monoacylated and 14 were diacylated. Non‐acylated flavonol glycosides were present as mono‐, di‐, tri‐ and tetraglycosides. This is the first time that the occurrence of four different fragmentation patterns of non‐acylated flavonol triglycosides has been reported in one matrix simultaneously. In addition, 44 flavonol glycosides were acylated with p‐coumaric, caffeic, ferulic, hydroxyferulic or sinapic acid. While monoacylated glycosides existed as di‐, tri‐ and tetraglycosides, diacylated glycosides occurred as tetra‐ and pentaglycosides. To the best of our knowledge, 28 compounds in kale are reported here for the first time. These include three acylated isorhamnetin glycosides (isorhamnetin‐3‐O‐sinapoyl‐sophoroside‐7‐O‐D‐glucoside, isorhamnetin‐3‐O‐feruloyl‐sophoroside‐7‐O‐diglucoside and isorhamnetin‐3‐O‐disinapoyl‐triglucoside‐7‐O‐diglucoside) and seven non‐acylated isorhamnetin glycosides. Copyright © 2010 John Wiley & Sons, Ltd.     

Determination of Melamine and Derivatives in Foods by Liquid Chromatography Coupled to Atmospheric Pressure Chemical Ionization Mass Spectrometry and Diode Array Detection

Two liquid chromatographic methods based on atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS) and diode array detection (DAD) are evaluated for the rapid determination of melamine (MEL) and structurally related compounds, including ammeline (AMN), ammelide (AMD), and cyanuric acid (CA) in foods. Both procedures used ion-exchange LC and isocratic elution. Samples were extracted by homogenization with acetonitrile/water/diethylamine. Specificity was demonstrated for LC-MS by the retention characteristics and MS spectra, by comparing with commercial standards. Specificity was only demonstrated in the case of LC-DAD for MEL and AMN, considering the retention characteristics and UV spectra. The recoveries obtained for spiked samples were satisfactory for all the analytes with LC-MS. The proposed procedure, LC-APCI-MS, was successfully applied to the analysis of different baby foods, including infant formula and breakfast cereal, and samples of rice flour, potato starch, soya drink, and coconut drink.     

基于实验和密度泛函理论研究EGCG与Zn(II)的相互作用

用紫外-可见(UV-Vis)吸收光谱和1H核磁共振(NMR)谱研究了茶多酚类衍生物表没食子儿茶素没食子酸酯(EGCG)与Zn(Ⅱ)离子的相互作用,并用密度泛函理论(DFT)计算了EGCG与Zn(Ⅱ)离子络合前后的空间结构及其紫外和核磁共振谱.实验与理论研究结果表明:EGCG主要构象是其芳香B环以e键(平伏键)及芳香D环以a键(直立键)形式共同与C环链接.EGCG通过其芳香D环上酚羟基与Zn(Ⅱ)离子相互作用,生成稳定的Zn(Ⅱ)与EGCG摩尔比为1:1的Zn(Ⅱ)-EGCG四面体络合物.     

Integrated system for on-line gas and liquid chromatography with a single mass spectrometric detector for the automated analysis of environmental samples

An integrated system has been developed which combines liquid (LC) and gas (GC) chromatographic separation with a single mass spectrometer (MS). On-line solid-phase extraction (SPE) of 10–200 ml aqueous samples on a short (10 × 2.0 mm I.D.) precolumn packed with a styrene-divinylbenzene copolymer is used for analyte enrichment. The trace-enrichment procedure was automated by means of a PROSPEKT cartridge-exchange/solvent-selection/valve-switching unit. After sample loading, the precolumn is eluted on-line in two subsequent runs, first onto the GC-MS system and, next, onto the LC-MS system using a particle beam (PB) interface. Prior to entering the PB-MS, the LC eluent passes through the flow cell of a UV diode-array detector (DAD). Both GC-MS and LC-PB-MS generate classical electron ionisation (EI) and chemical ionisation (CI) spectra which are useful for the identification of low- and sub-μg/l concentrations of environmental pollutants covering a wide polarity and volatility range. The LC-DAD data provide additional means for quantitation and yield complementary spectral information. All three detection systems (GC-MS, LC-DAD, LC-PB-MS) and the trace-enrichment procedure are fully automated and controlled from the keyboard of the central computer. With such a ‘MULTIANALYSIS’ system GC-MS, LC-DAD and LC-MS data of the same sample can be obtained within 3 h. The system was optimised with nine chlorinated pesticides in drinking water as test mixture. With 100-ml samples detection limits in GC-MS were 0.0005−0.03 μg/l, and in LC-PB-MS 0.5–7 μg/l, both in the full-scan (EI) mode. Negative chemical ionisation (NCI) with methane as reagent gas improved the sensitivity of six halogenated compounds 3- to 30-fold and provided relevant information for structural elucidation of unknown compounds in real-world samples. LC-DAD detection limits varied from 0.01 to 0.05 μg/l. Relative standard deviations (R.S.D.) of retention times were less than 0.2% in all systems, R.S.D.s of peak areas were 5–15% for GC-MS and LC-PB-MS and less than 5% for LC-DAD. The ‘MULTIANALYSIS’ system was used to analyse surface water samples and river sediment extracts; several pollutants were detected and identified.     

Simultaneous quantification of terpenelactones and flavonol aglycones in hydrolyzed ginkgo biloba extract by liquid chromatography with inline ultraviolet and evaporative light scattering detection

We report here a liquid chromatography (LC) method with inline ultraviolet/evaporative light scattering (UV/ELS) detection for the simultaneous quantification of the terpenelactones and flavonol aglycones in a single sample of hydrolyzed Ginkgo biloba extract (GBE). The sample is hydrolyzed by a rapid and convenient oven heating method for 1 h at 90 degrees C with 10% hydrochloric acid. The 1 h hydrolysis was found to be equivalent to the 2.25 h reflux treatment for dry powder extract, where total flavonol glycosides were 28.4 and 28.1%, respectively. Acceptable precision was achieved for total terpenelactones [relative standard deviation (RSD) = 4.8%] by ELS detection, and total flavonol aglycones (RSD = 2.3%) by UV detection. The analytical range was 1.5 to 7.3% (w/w) for the individual terpenelactones (ELS) and 2.5 to 15.0% (w/w) for the individual glycosides (UV) calculated from the aglycones quercetin, kaempferol, and isorhamnetin. This improved method allows for the first time high throughput sample preparation coupled with the quantification of the predominant compounds generally used for quality control of GBE in a single assay.     

Determination of absolute configuration of axially chiral biaryls

Absolute configurations of axially chiral biaryls with hydroxyl, amino, and carboxyl groups can be easily determined by pmr spectra using MTPA derivatives and shift reagents.     

Nuclear magnetic resonance and liquid chromatography-mass spectrometry combined with an incompleted separation strategy for identifying the natural products in crude extract

NMR and LC-MS combined with an incompleted separation strategy were proposed to the simultaneous structure identification of natural products in crude extracts, and a novel method termed as NMR/LC-MS parallel dynamic spectroscopy (NMR/LC-MS PDS) was developed to discover the intrinsic correlation between retention time (Rt), mass/charge (m/z) and chemical shift (δ) data of the same constituent from mixture spectra by the co-analysis of parallelly visualized multispectroscopic datasets from LC-MS and ¹H NMR. The extracted ion chromatogram (XIC) and ¹H NMR signals deriving from the same individual constituent were correlated through fraction ranges and intensity changing profiles in NMR/LC-MS PDS spectrum due to the signal amplitude co-variation resulted from the concentration variation of constituents in a series of incompletely separated fractions. NMR/LC-MS PDS was applied to identify 12 constituents in an active herbal extract including flavonol glycosides, which was separated into a series of fractions by flash column chromatography. The complementary spectral information of the same individual constituent in the crude extract was discovered simultaneously from mixture spectra. Especially, two groups of co-eluted isomers were identified successfully. The results demonstrated that NMR/LC-MS PDS combined with the incompleted separation strategy achieved the similar function of on-line LC-NMR-MS analysis in off-line mode and had the potential for simplifying and accelerating the analytical routes for structure identification of constituents in herbs or their active extracts.     

Ultrasound-assisted extraction of Cordyceps cicadae polyphenols: Optimization,LC-MS characterization,antioxidant and DNA damage protection activity evaluation

In this study, ultrasound-assisted extraction of polyphenols from C. cicadae was optimized by response surface methodology (RSM). The optimized conditions were determined as extraction time of 39 min, liquid-to-solid ratio of 1:29 g/mL, extraction temperature of 69 °C and ethanol concentration of 55% with a yield of 21.9 mg gallic acid equivalent/g dry weight. Four resins were used for polyphenol purification. D101 resin had the highest ratio of adsorption and was further applied in polyphenol purification test. A total of 19 different phenolic compounds were identified by LC-MS, including 12 phenolic acids and 7 organic acids. In addition, C. cicadae polyphenols displayed higher antioxidant activity in vitro and anti-aging activity of C. elegans in vivo. Lastly, C. cicadae polyphenols showed the potential to protect DNA from oxidative damage. Overall, our results suggest that polyphenols from C. cicadae may be considered as novel sources of anti-oxidation, anti-aging and recommended as reagents to protect DNA from oxidative damage in food and pharmaceutical industries.     

Analysis of p-chlorobenzoic acid in water by liquid chromatography-tandem mass spectrometry

para-Chlorobenzoic acid (p-CBA) is typically used as a probe compound to indirectly quantify hydroxyl radicals formed during advanced oxidation processes used in drinking water and wastewater treatment. A method has been developed for the sensitive analysis of p-CBA in water using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A reporting limit in water of 100 ng/L was determined for the method, which is 40-fold lower than the 4.0 microg/L reporting limit of the widely used liquid chromatography with UV detection (LC-UV) method. The method was found to be robust in difficult matrices such as wastewater and highly selective, unlike LC-UV which relies on non-specific detection at 234 nm. The detection of p-CBA below 1 microg/L during bench-scale ozonation of wastewater after hydrogen peroxide addition was demonstrated. Duplicate samples were analyzed by LC-MS/MS and LC-UV and results were found to be comparable at concentrations quantifiable by both methods.     

Synthesis of imidazole‐containing conjugated polymers bearing phenol unit as side group and excited state intramolecular proton transfer‐mediated fluorescence

Dibromobenzimidazole and dibromoimidazole bearing hydroxyl group‐protected phenol unit ( 1 and 2 ) were prepared and they showed an intramolecular hydrogen bonding between ether oxygen and amino proton of imidazole. The palladium‐catalyzed Suzuki coupling polymerization of 1 and 2 with benzene bis(boronic acid) derivatives gave soluble polymers ( 3 and 4 ), where the molecular weights were limited probably due to the coordination ability of imidazole to palladium metal. The phenol hydroxyl groups were subsequently deprotected using BBr₃ to obtain 3 ′ and 4 ′. From the ¹H NMR spectra, the complete conversion to the hydroxyl group and the formation of another type of intramolecular hydrogen bonding between hydroxyl proton and imine nitrogen were confirmed. In the UV and PL spectra of 3 ′ and 4 ′, the excited state intramolecular proton transfer (ESIPT) occurred to shift the emission spectra toward lower energy region compared to 3 and 4 . Especially, the PL spectrum of 3 ′ demonstrated large stokes shift (145 nm) in THF solution. The ESIPT‐mediated fluorescence was influenced by the addition of methanol and trifluoroacetic acid, which inhibited the formation of intramolecular hydrogen bonding. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 4822–4829, 2009     

Kaempferol tri- and tetraglycosides from the flowers of Clematis cultivar

A new kaempferol glycoside, kaempferol 3-O-alpha-rhamnopyranosyl-(1 --> 2)-[alpha-rhamnopyranosyl-(1 --> 6)-beta-glucopyranoside]-7-O-beta-glucopyranoside (2) was isolated from the flowers of Clematis cultivar "Jackmanii Superba", together with a known kaempferol 3-O-alpha-rhamnopyranosyl-(1 --> 6)-beta-glucopyranoside-7-O-beta-glucopyranoside (1). The chemical structures of the isolated glycosides were established by UV, LC-MS, characterization of acid hydrolysates, and 1H and 13C NMR spectroscopy.     

油菜蜂花粉黄酮含量的HPLC测定

以95％乙醇为溶剂,采用索氏提取器提取青海产油菜蜂花粉中的黄酮类化合物,将黄酮提取物中的黄酮甙水解为黄酮甙元后,利用HPLC法测定其中槲皮素、山萘酚、异鼠李素含量。结果表明,青海油菜蜂花粉中槲皮素、山萘酚、异鼠李素的平均含量分别为0．928％、0．295％、0．0834％,换算成总黄酮含量为3．28％。     

Immobilized triazine-type dehydrocondensing reagents for carboxamide formation: ROMP-Trz-Cl and ROMP(OH)-Trz-Cl

New triazine-type dehydrocondensing reagents, such as ROMP-Trz-Cl and ROMP(OH)-Trz-Cl, were synthesized by a ring opening metathesis polymerization (ROMP) method, and these showed higher loading than conventional polymer-supported condensing reagents. These polymers effect the formation of amides in good yields by addition of a mixture of carboxylic acid, amine and NMM. ROMP(OH)-Trz-Cl, which contains hydroxyl groups in the polymer chain, gave amides in good yields even in MeOH.     

Combined mass spectrometric and chromatographic methods for in‐depth analysis of phenolic secondary metabolites in barley leaves

Structural analysis via HPLC‐ESI‐MSn, UPLC‐HESI‐MS/MS and NMR reported 152 phenolic secondary metabolites in spring barley seedlings (Hordeum vulgare L.). Flavonoids with various patterns of glycosylation and acylation, as well as hydroxycinnamic acid glycosides, esters and amides, were identified in methanolic extracts from leaves of nine varieties of barley originating from different regions of the world. Hordatines derivatives, flavones acylated directly on the aglycone, and hydroxyferulic acid derivatives deserve special attention. Preparative chromatography enabled characterization of a number of compounds at trace levels with the 6‐C‐[6″‐O‐glycosyl]‐glycosides and the 6‐C‐[2″,6″‐di‐O‐glycosides]‐glucoside structure of flavones. Derivatives of flavonols, quercetin and isorhamnetin were observed only in Syrian varieties. The ultra performance liquid chromatography profiles of UV‐absorbing secondary metabolites were used for chemotaxonomic comparison between nine varieties of barley from different climatic conditions. The hierarchical clustering of bred lines from the Fertile Crescent and European and American varieties indicates a great diversity of chemical phenotypes within barley species. Copyright © 2015 John Wiley & Sons, Ltd.     

Photooxidation of poly[methyl(phenyl)silylene] and effect of photostabilizers

Poly[methyl(phenyl)silylene] (PMPSi) samples were irradiated in air atmosphere using either a weatherometer ATLAS Ci 3000+ or a mercury discharge lamp HBO 200. The kinetics of photodegradation was determined by FTIR and UV spectroscopies. The absorption changes in the siloxane, carbonyl and hydroxyl regions were monitored and analyzed. It is assumed that photodegradation of neat PMPSi in air atmosphere is an irreversible dual photoprocess consisting of primary photolysis followed by the photooxidation to oxygenated products such as siloxanes and species containing hydroxyl and carbonyl groups. Degradative changes can be retarded by the addition of photostabilizers. Triazine-based phenolic UV absorber protecting the polymer by the excited-state intramolecular proton transfer mechanism, bifunctional stabilizer (consisting of UV absorber and HAS functions) and the combination of triazine-based phenolic UV absorber with HAS was rather effective in retardation of photodegradation of PMPSi.