•A new HPLC-MS/MS method for analysis of 36 phenolics in berry fruits and jams.•Blueberry exhibited higher contents of anthocyanins, flavonols and phenolic acids.•Strawberry showed higher amounts of ...flavan-3-ols, dihydrochalcones and flavanones.•Higher phenolic content in the blueberry justified greater antioxidant activity.•This HPLC-MS/MS method is reliable for quality control analyses of fruit products.
Berry fruits consumption has increased in recent years because they are rich sources of polyphenols with reported health benefits. The aim of the present work was to develop a new comprehensive and fast HPLC-MS/MS method for simultaneous determination of 36 phenolic compounds (7 anthocyanins, 9 flavonols, 4 flavan-3-ols, 2 dihydrochalcones, 2 flavanones and 12 phenolic acids) present in blueberry, strawberry, and their fruit jam. Blueberry fruits showed higher contents of anthocyanins, flavonols and phenolic acids, while strawberry fruits exhibited higher contents of flavan-3-ols, dihydrochalcones and flavanones. Anthocyanins were the main phenolic constituents in both berries. Furthermore, the higher total phenolic content in the blueberry fruit and jam justified their greater antioxidant capacity measured by DPPH free radical assay, compared to strawberry. In conclusion, this new HPLC-MS/MS method is useful and reliable for quality control and authentication analyses of blueberry and strawberry fruits and their commercial food products, such as jams.
•The antioxidant activity of XHP was evaluated.•XHP were separated based on molecular weight and polarity differences.•Peptide with the strongest activity had 74.45% scavenging effect on DPPH.•The ...sequence of main antioxidative peptide was identified as Asp-Leu-Glu-Glu.
This study mainly focused on the purification and identification of antioxidative peptides generated in dry-cured Xuanwei ham. Based on scavenging effect on free radicals and ferrous ion, the antioxidant activity of crude peptides from Xuanwei ham was assessed. From the scavenging effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl (OH) radicals and superoxide anion (O2−), it was suggested that XHP generated during the ripening period had a strong antioxidant activity. By using size exclusion chromatography, anion exchange column and reversed-phase HPLC, fractions with a strong antioxidative activity were separated based on their molecular weight and polarity differences. The fraction with strong antioxidant effect was further characterized by LC–MS/MS. The results suggest that antioxidative peptides are produced during the long processing of Xuanwei ham among which the tetrapeptide Asp-Leu-Glu-Glu could be one of the main peptides that play key role in the antioxidant activity.
The distribution of betalains in peel, flesh, and petioles of yellow and red beetroot cultivars has been investigated using an High‐performance liquid chromatography (HPLC) system with electrospray ...mass spectrometry. Differences in the levels of betacyanins and betaxanthins between different colored cultivars were individually determined for 3 plant parts. The content of almost all analyzed compounds decreased in the following order: peel > flesh > petiole. Betanin/isobetanin pigments comprised a major portion of the relative peak area measured in red beetroot peel. Isobetanin relative peak areas were also high in leaf petioles (68.94% to 74.16%) of red colored cultivars. However, betacyanins were completely absent from the extracts of all analyzed parts of yellow beet. Glutamine‐bx represented a very high relative peak area (59.54% to 64.18%) in flesh of red‐colored cultivars analyzed in the study. Our results indicate that red beet cultivars can be utilized as a potential source of red and yellow natural colorants. However, differences in pigment composition among different beetroot parts must be considered and in order to maximize the pigment yields petioles can also be used as a source rich in specific betalain compounds.
Practical Application
The studies on plant material with high betalain levels can provide new opportunities for the use of these healthy pigments in the food and pharmaceutical industries. Further investigations on highly concentrated betalains in various cultivars/plants parts can help elucidate their biological activity.
•A low-sample-volume SPE-HPLC–MS/MS method was developed.•Antibiotics, metabolites and tracers were separated by RP and HILIC.•The method was established for long-term monitoring of an urban sewage ...treatment plant.•Recovery rates ranged between 51% and 111%.•The method is simple, fast and easily applicable to other sewage monitorings.
A rapid analytical method was developed for the application of a long-term monitoring (>one year) of the most prescribed and often in hospitals used antibiotics in diverse wastewaters of an urban sewage treatment plant (STP). Additionally to the selected multi-class antibiotics amoxicillin, penicillin V and piperacillin (penicillins), cefotaxime and cefuroxime (cephalosporins), azithromycin, clarithromycin and roxithromycin (macrolids), ciprofloxacin and levofloxacin–ofloxacin (fluoroquinolones), clindamycin (lincosamide), doxycycline (tetracycline), sulfamethoxazole (sulfonamide) and trimethoprim (dihydrofolate reductase inhibitor), the bioactive metabolite clindamycin-sulfoxide, the reserve antibiotic vancomycin (glycopeptide) and as tracer of the STP the anticonvulsant carbamazepine and the antifungal fluconazole were involved. The analytical method combines a low-sample-volume solid phase extraction (SPE), followed by a chromatographic separation using a reversed phase (RP) and hydrophilic interaction liquid chromatography (HILIC) technique, respectively, coupled to a triple quadrupole mass spectrometer. Detection was performed with multiple reaction monitoring (MRM) measured with positive electrospray ionization (ESI+). The extraction efficiency of different SPE cartridges and optimized pH-values of the preparation procedure were tested. Finally, the extraction of antibiotics was realized with the Oasis HLB cartridge and a pH adjustment at 3.5. An external calibration curve in diluted blank urine was used for quality control of the sample set of daily composite samples of the STP for the duration of one year monitoring. The squared coefficient of determination (r2) in the concentration range (20–20,000ng/L or 100–100,000ng/L) of the calibration curves for the method was higher than 0.99 for all determined substances. The limit of quantification (LoQ) ranged between 0.8ng/L (azithromycin) and 245.1ng/L (vancomycin). Furthermore, a standard addition was used for quantification in wastewater samples. The process efficiencies ranged from 20% (doxycycline) to 134% (cefuroxime) in influent samples and from 31% (doxycycline) to 171% (cefuroxime) in effluent samples of the STP. All selected substances have been found in wastewater samples. Cefuroxime, doxycycline, levofloxacin, piperacillin, sulfamethoxazole and carbamazepine showed highest concentrations up to 6.2μg/L.
Characterization of highly glycosylated biopharma-ceuticals by mass spectrometry is challenging because of the huge chemical space of coexistent glycoforms present. Here, we report the use of an ...array of HPLC-mass spectrometry–based approaches at different structural levels of released glycan, glycopeptide, and hitherto unexplored intact glycoforms to scrutinize the biopharmaceutical Myozyme, containing the highly complex lysosomal enzyme recombinant acid α-glucosidase. The intrinsic heterogeneity of recombinant acid α-glucosidase glycoforms was unraveled using a novel strong anion exchange HPLC-mass spectrometry approach involving a pH-gradient of volatile buffers to facilitate chromatographic separation of glycoforms based on their degree of sialylation, followed by the acquisition of native mass spectra in an Orbitrap mass spectrometer. Upon considering the structures of 60 different glycans attached to seven glycosylation sites in the intact protein, the large set of interdependent data acquired at different structural levels was integrated using a set of bioinformatic tools and allowed the annotation of intact glycoforms unraveling more than 1,000,000 putative intact glycoforms. Detectable isoforms also included several mannose-6-phosphate variants, which are essential for directing the drug toward its target, the lysosomes. Finally, for the first time, we sought to validate the intact glycoform annotations by integrating experimental data on the enzymatically dissected proteoforms, which reduced the number of glycoforms supported by experimental evidence to 42,104. The latter verification clearly revealed the strengths but also intrinsic limitations of this approach for fully characterizing such highly complex glycoproteins by mass spectrometry.
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•Hybrid mass spectrometry approach to characterize Myozyme at the different structural levels.•Intact Myozyme analysis by a novel strong anion exchange-HPLC-mass spectrometry approach enabling the separation of proteoforms based on their sialylation degree and acquisition of native mass spectra in a semi-automated fashion.•Enzymatic dissection with peptide:N-glycosidase F and sialidase to reduce spectral complexity at intact protein level.•Multilevel data integration using the software MoFi to annotate intact protein mass spectra.•Application of the approach to calculate the percentage of the biologically relevant phosphorylated glycoforms.
Compared to the total number of mammalian genes, the structural diversity of glycoproteins as a consequence of posttranslational glycosylation is enormous. We propose an integrated HPLC-mass spectrometry approach to explore the highly glycosylated protein recombinant acid alpha-glucosidase (Myozyme). The glycosylation complexity of this protein is reflected in a huge chemical space of more than 40,000 glycoforms, which were experimentally revealed through HPLC-mass spectrometry analyses and bioinformatic data integration at different structural levels of released glycans, glycopeptides, and intact glycoforms.
HPLC-MS-based methods for the study of metabonomics Wilson, Ian D; Plumb, Robert; Granger, Jennifer ...
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences,
03/2005, Volume:
817, Issue:
1
Journal Article
Peer reviewed
The development and use of HPLC-MS for the study of metabonomics is reviewed. To date the technique has been applied to the analysis of urine samples obtained from studies in rodents in ...investigations of physiological variation (e.g., factors such as strain, gender, diurnal variation, etc.) and toxicity. Examples are provided of the use of conventional HPLC, capillary methods and the recently introduced high-resolution systems based on a combination of high pressure and small particle size ("UPLC"). Comparison is also made of the use of 1H NMR spectroscopy and HPLC-MS for the analysis of biofluid samples and the advantages and limitations of the two approaches are assessed. Likely future developments are considered.
•Optimization of chestnut shells extraction using response surface methodology.•Subcritical water extraction is a clean method for antioxidants extraction.•Optimum extraction conditions: temperature ...220 °C and extraction time 30 min.•Caffeoylquinic acid isomers were the main phenolic compounds.
The objective of this study was to evaluate the optimal Subcritical Water Extraction (SWE) conditions of antioxidants and polyphenols from chestnut shells using Response Surface Methodology (RSM). A central composite design (CCD) was conducted to analyse the time (6–30 min) and temperature (51–249 °C) effects in antioxidant activity (ABTS, DPPH and FRAP) and Total Phenolic Compounds (TPC). TPC ranged from 315.21 to 496.80 mg gallic acid equivalents (GAE)/g DW; the DPPH from 549.23 to 1125.68 mg Trolox equivalents (TE)/g DW; ABTS varied between 631.16 and 965.45 mg ascorbic acid equivalents (AAE)/g DW and FRAP from 2793.95 to 11393.97 mg ferrous sulphate equivalents (FSE)/g DW. The optimal extraction conditions were 30 min/220 °C, revealing excelling scavenging efficiencies against HOCl (IC50 = 0.79 µg/mL) and O2− (IC50 = 12.92 µg/mL) without toxicity on intestinal cells (0.1 µg/mL). The phenolic composition revealed high amounts of pyrogallol and protocatechuic acid. SWE can be a useful extraction technique for the recovery of polyphenolics from chestnut shells.
•Green synthesis of bimetallic Fe/Pd NPs was firstly reported using the one-step method.•98.0% of Orange II was removed by Fe/Pd NPs, but only 16.0% by Fe NPs.•Fe/Pd NPs with a diameter ranging from ...10 to 100nm were observed.•Removing Orange II using Fe/Pd NPs involved both adsorption and catalytic degradation.
To reduce cost and enhance reactivity, bimetallic Fe/Pd nanoparticles (NPs) were firstly synthesized using grape leaf aqueous extract to remove Orange II. Green synthesized bimetallic Fe/Pd NPs (98.0%) demonstrated a far higher ability to remove Orange II in 12h compared to Fe NPs (16.0%). Meanwhile, all precursors, e.g., grape leaf extract, Fe2+ and Pd2+, had no obvious effect on removing Orange II since less than 2.0% was removed. Kinetics study revealed that the removal rate fitted well to the pseudo-first-order reduction and pseudo-second-order adsorption model, meaning that removing Orange II via Fe/Pd NPs involved both adsorption and catalytic reduction. The remarkable stability of Fe/Pd NPs showed the potential application for removing azo dyes. Furthermore, Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) confirmed the changes in Fe/Pd NPs before and after reaction with Orange II. High Performance Liquid Chromatography–Mass Spectrum (HPLC–MS) identified the degraded products in the removal of Orange II, and finally a removal mechanism was proposed. This one-step strategy using grape leaf aqueous extract to synthesize Fe/Pd NPs is simple, cost-effective and environmentally benign, making possible the large-scale production of Fe/Pd NPs for field remediation.
•The anthocyanin composition six Chilean small berries were investigated.•Analysis was performed using high resolution mass analysis (HR-ToF-ESI-MS).•Compounds were mostly branched 3-O-glycosides of ...cyanidin and malvidin.•Thirty one anthocyanins, three phenolic acids and five flavonols were identified.•Calafate showed the highest antioxidant activity followed by blueberry and arrayán.
The HPLC profiles of six fruits endemic of the VIII region of Chile were investigated using high resolution mass analysis (HR-ToF-ESI-MS). The anthocyanin fingerprints generated for the fruits were compared and the antioxidant capacities measured by the scavenging of the DPPH radical, the ferric reducing antioxidant power (FRAP), the superoxide anion scavenging activity assay (SA), and correlated with the inhibition of lipid peroxidation in human erythrocytes (LP) and total content of phenolics, flavonoids and anthocyanins measured by spectroscopic methods. Several anthocyanins were identified, including 3-O-glycosides derivatives of delphinidin, cyanidin, petunidin, peonidin and malvidin. Three phenolic acids (feruloyl-quinic acid, chlorogenic acid, and neochlorogenic acid) and five flavonols (hyperoside, isoquercitrin, quercetin, rutin, myricetin and isorhamnetin) were also identified. Calafate fruits showed the highest antioxidant activity. However, the highest LP activity was found for Chilean blueberries (>95%) followed by calafate fruits (91.27%) and luma (83.4%).
Highlights: • Antioxidant activity of skin and flesh of different coloured onions were studied. • Phenolic content of onion skin was higher than flesh; Pearl skin containing the highest. • ...Antioxidant assays showed a similar trend; Pearl and Red skin being the best. • HPLC analysis showed quercetin glucoside, quercetin and kaempferol in onion extracts. Abstract: The antioxidant activity of phenolic constituent of skin and selected flesh of different coloured onions (Pearl, Red, Yellow and White) were determined. The green shoot obtained after sprouting of the red onion was also analyzed to study the changes in the phenolic constituents during germination. For the first time, all tests were carried out separately for the free, esterified and insoluble-bound phenolic constituents of onion samples. The content of phenolics extracted from onion skins was approximately six times higher than that of their flesh counterparts. Among onion varieties, pearl onion skin showed the highest phenolic content (26.4 mg quercetin eq/g freeze dried sample). A similar trend was observed for free radical scavenging activity of the tested samples. Extracts from edible part of onion showed lower activity in all antioxidant tests carried out. The HPLC–MS analysis showed that quercetin 3,4′-diglucoside, quercetin, and kaempferol were the predominant phenolics in all onion extracts tested.
