Characterization of phenolic extracts from olives (Olea europaea cv. Pisciottana) by electrospray ionization mass spectrometry

Olives are a potential source of natural anti-oxidants; unfortunately, a complete characterization of olive fruit phenolic profile is still to be achieved, as it is characterized by great complexity and variability.     

Investigation of proanthocyanidins by HPLC with electrospray ionization mass spectrometry

Received: 1 June 1999     

Antioxidant activity of Annona crassiflora: Characterization of major components by electrospray ionization mass spectrometry

The polar components of Annona crassiflora pulp, peel and seeds ethanolic extracts were investigated by direct infusion electrospray ionization mass spectrometry (ESI-MS) both in the negative ion mode. Characteristic ESI mass spectra with many diagnostic ions were obtained for the extracts, serving for fast and reliable information. The technique provided information of component structures revealing the presence of important bioactive components widely reported as potent antioxidants such as ascorbic acid, caffeic acid, quinic acid, ferulic acid, xanthoxylin, rutin, caffeoyltartaric acid, caffeoyl glucose and [quercetin+hexose+pentose−H]⁻¹ This is the first report on the composition by ESI-MS of araticum peel and seed ethanolic extracts demonstrating excellent antioxidant activity.     

Electrospray ionization mass spectrometry fingerprinting of essential oils: Spices from the Labiatae family

Polar components of the methanolic extracts of the essential oils of the spices Origanum dictamnus, Origanum vulgare, Origanum majorana and Rosmarinus officinalis, all four belonging to the Labiatae family, were investigated by direct infusion electrospray ionisation mass spectrometry (ESI-MS) both in the negative and positive ion modes. Characteristic ESI mass spectra with many diagnostic ions were obtained for the extracts of all four spices, serving for fast and reliable identification of these species. Tandem mass spectrometry (ESI-MS/MS), which often forms a series of fragment ions, and this additional MS dimension increases selectivity for authenticity and adulteration tests for spice essential oils. The MS technique also provides complementary information of component structures revealing the presence of important bioactive components.     

Antioxidant activity of Caryocar brasiliense (pequi) and characterization of components by electrospray ionization mass spectrometry

The Caryocar brasiliense known commonly as pequi is a tropical fruit of Brazilian Cerrado and is considered an important option of income and food for the populations living in this biome. Our previous study indicated that C. brasiliense had high total phenol content (209 g as gallic acid equivalent kg⁻¹) and excellent scavenging activity against 2,2-diphenyl-1-picrylhydrazyl radical (IC₅₀ of 9.44 μg ml⁻¹). In this study, we evaluated the highly efficient antioxidant activity of C. brasiliense using the biological relevant method of chemically induced lipid peroxidation. The half inhibition concentration did not exceed 0.8 μg ml⁻¹. In addition, polar components of pequi ethanolic extract were investigated by direct infusion electrospray ionization mass spectrometry (ESI-MS). The technique revealed the presence of important bioactive components widely reported as potent antioxidants such as gallic acid, quinic acid, quercetin, and quercetin 3-O-arabinose possibly explaining its higher antioxidant activity. This is the first report on the composition by ESI-MS of pequi extract demonstrating excellent antioxidant activity.     

Molecular mass distribution of dextran in Brazilian sugar and insoluble deposits of cachaça

The dextran molecular mass distribution profile in 77 sugar samples from Brazil and twelve insoluble deposits (alcoholic flocks) samples from sugared cachaças (Brazilian sugar cane spirit) is described in terms of number-average molecular mass M_n, weight-average molecular mass M_w, Z-average molecular mass M_z, and polydispersity. The analyses were performed by size-exclusion chromatography, using a refractive index detector. In most of the sugar samples, it was possible to identify two major groups of dextrans with M_w averages of 5 × 10⁶ and 5 × 10⁴ Da. Based on the evaluated parameters, the dextran distribution profile is about the same in samples analyzed over five seasons, and, therefore, it is likely that the Brazilian product pattern will not change very much over the years. In insoluble deposits from sugared cachaças, dextrans with M_w values in the order of the 10⁵ Da were the most frequent ones, being present in 58% of the samples.     

Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs

In this study, we have demonstrated an accurate and rapid small RNA analytical method with both sequence determination and detailed modification analysis by electrospray ionization-ion trap/time-of-flight mass spectrometry (ESI-IT/TOFMS). To develop this ideal method, we have examined the performance of ESI-IT/TOFMS using various chemically-synthesized model sequences of modified or unmodified microRNAs (miRNAs). The deconvoluted mass of a 22-nucleotide (nt) miRNA was obtained from a multiply charged precursor ion (MS(1)). The ion exhibited high mass accuracy (< 7 ppm) and high mass resolution (a value of m/Δm=10,000) and was therefore very useful in RNA composition assignment. The optimized MS(2) method using ion trap collision-induced dissociation, as well as automatic annotation analysis of product ions based on the accurate mass information, enabled the precise sequencing determination of intact miRNAs. Further, the detailed structural analysis of 3'-terminal modified nucleic acid in intact methylated miRNA was carried out using the MS(3) capability of the hybrid IT/TOFMS. The direct infusion method also provided a high throughput and good sensitivity because the analytical time and sample concentration needed in a series of experiments with reliable data were only 3 min and 100 nM, respectively. This study provides a novel approach for characterizing the intact chemically-synthesized small RNA without chemical and enzymatic digestions and would be widely applicable for the structural analysis of complicated modified small RNAs.     

Isolation and identification of antioxidative peptides from porcine collagen hydrolysate by consecutive chromatography and electrospray ionization–mass spectrometry

The porcine skin collagen was hydrolyzed by different protease treatments to obtain antioxidative peptides. The hydrolysate of collagen by cocktail mixture of protease bovine pancreas, protease Streptomyces and protease Bacillus spp. exhibited the highest antioxidant activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, metal chelating and in a linoleic acid peroxidation system induced by Fe²⁺. And degree of hydrolysis highly affected the antioxidant properties of the hydrolysates. Four different peptides showing strong antioxidant activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration chromatography, ion-exchange chromatography and high-performance liquid chromatography. The molecular masses and amino acid sequences of the purified antioxidant peptides were determined using electrospray ionization (ESI) mass spectrometry. One of the antioxidative peptides, Gln-Gly-Ala-Arg, was then synthesized and the antioxidant activities measured using the aforementioned methods. The results confirmed the antioxidant activity of this peptide, and adds further support to its feasibility as a provider of natural antioxidants from porcine skin collagen protein.     

C NMR and electrospray ionization mass spectrometric study of sucrose aqueous solutions at high pH: NMR measurement of sucrose dissociation constant

The ¹³C NMR technique is used for the measurement of the first dissociation constant of sucrose (HL) in highly alkaline solutions. In 1.0 M NaCl/NaOH medium and for 25 °C, the concentration dissociation constant (pK₁) was 13.1 ± 0.3; and, for 60 °C, pK₁ = 12.30 ± 0.05. The β-d-fructofuranosyl ring was found to be responsible for dissociation. The NMR data reveal no clear evidence of the second dissociation step below pH 14, either at 25 °C or at 60 °C. In the solutions with 4–10 mol dm⁻³ NaOH content the ¹³C NMR technique indicated the chemical shift changes, treated as the second dissociation step of sucrose and a sodium complex formation. A very rough estimation, for variable ionic strength, gives the value: pK₂ ∼ 15.8 ± 0.8. The anionic species L⁻ and NaH₋₁L⁻ have been registered by electrospray ionization time-of-flight mass spectrometry (ESI-ToF MS) for 0.01 M sucrose solutions with initial pH 13.     

Ethyl carbamate in pot still cachaças (Brazilian sugar cane spirits): Influence of distillation and storage conditions

Ethyl carbamate (EC), a known genotoxic carcinogen, was studied in 25 brands of pot still cachaças from 19 distilleries in Paraíba State, Brazil. A concentration range of 55–700 μg/l was found with most brands (∼70%) exceeding the international EC limit for spirits (150 μg/l). Brand characteristics (colour, distillation [single or double], and bottle colouration) showed no consistent connection with EC levels. However, when EC levels of yellowish (cask matured) and colourless single-distiled cachaças from the same distillery were compared, the yellowish type was much more heavily contaminated. Eleven distilleries were visited and information regarding the distillation scale, kettle heating system, kettle’s shape, and cooling system of the column was collected. A close connection between EC levels and cooling system was found, with the non-cooled and cooled columns dominating the brands with high (200–700 μg/l range) and low (55–100 μg/l range) contamination levels, respectively.     

Studies on structures and activities of initial Maillard reaction products by electrospray ionisation mass spectrometry combined with liquid chromatography in processing of red ginseng

Electrospray ionisation-mass spectrometry (ESI-MS) was applied to analyse the water-soluble extract of red ginseng (RG). Several new compounds were produced from the Maillard reaction during the steaming and drying process for preparing RG. Both the tandem electrospray ionisation (ESI-MS(n)) and Fourier transform ion cyclotron resonant mass spectrometric (FT-ICR-MS) data of these products proved that they were the initial Maillard reaction products (MRPs) of maltose with glutamic acid/aspartic acid, which were specific components in RG. In addition, their anti-diabetic and antioxidant activities were examined in vitro. The anti-diabetic activities were evaluated by studying the α-glucosidase inhibition using ultrafiltration LC-MS/MS techniques, while the antioxidant activities were investigated by UPLC-ESI-MS method. The results demonstrated that four initial MRPs in RG were identified as α-glucosidase inhibitors, and showed marked scavenging effect on the hydroxyl radical ((·)OH). Based on these studies, the processing method of RG was improved to generate more active compounds.     

Determination of diquat and paraquat in olive oil by ion-pair liquid chromatography–electrospray ionization mass spectrometry (MRM)

For the first time a method for determination of herbicides diquat (DQ) and paraquat (PQ) in olive oil was developed utilising liquid chromatography–electrospray ionization mass spectrometry (MRM). n-Hexane/10 mM HFBA aqueous solution partitioning was used as the extraction method. Separation was carried out in an Xterra C8 column (100 × 21 mm, 3 μm), using the gradient mode. Solvent A was a HFBA aqueous solution (5 mM, pH 2) and solvent B acetonitrile/methanol 75/25 (v/v). Peaks used for quantification were m/z = 157 (diquat) and m/z = 158 (paraquat). Detection limit found for both diquat and paraquat was 4 μg kg⁻¹. The method can also be applied for determination of chlormequat (CQ, quantification peak m/z = 58), the detection limit being 0.3 μg kg⁻¹. Such limits are clearly lower than the MCLs commonly applied to olive oil as reference criteria (5 times MCLs in olives). Good reproducibilities (day to day and run to run) were obtained.     

Short communication: Evaluation of MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci

This study evaluated MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci (CNS). A total of 861 CNS isolates were used in the study, covering 21 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 804) and the remainder were identified by sequencing of hsp60 (n = 56) and tuf (n = 1). The genotypic identification was considered the gold standard identification. Using a direct transfer protocol and the existing commercial database, MALDI-TOF mass spectrometry showed a typeability of 96.5% (831/861) and an accuracy of 99.2% (824/831). Using a custom reference spectra expanded database, which included an additional 13 in-house created reference spectra, isolates were identified by MALDI-TOF mass spectrometry with 99.2% (854/861) typeability and 99.4% (849/854) accuracy. Overall, MALDI-TOF mass spectrometry using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS.     

Geochemical and instrumental fundamentals for accurate and precise strontium isotope data of food samples: Comment on “Determination of the strontium isotope ratio by ICP-MS ginseng as a tracer of regional origin” ()

The strontium isotope composition of food samples has been successfully used to trace the provenance of both herbal and animal agricultural products. Adequately accurate and precise ⁸⁷Sr/⁸⁶Sr ratios, however, are key to unravel environmental and anthropogenic strontium contributions of specific growing areas. In a recently published analytical method in Food Chemistry by Choi et al. (2008) a demonstrably inadequate protocol for the determination of ⁸⁷Sr/⁸⁶Sr ratios of ginseng is presented. Based on well established geological and analytical fundamentals the ⁸⁷Sr/⁸⁶Sr ratios presented by Choi et al. (2008) are too inaccurate and imprecise to be used to trace the origin of the ginseng. Appropriate wet chemical separation techniques and mass spectrometric instrumentation enables the generation of ⁸⁷Sr/⁸⁶Sr data with very low expanded uncertainties (<0.05%, k = 2). Those high quality data allow the identification of even small strontium contributions of source reservoirs or physical and anthropogenic processes if diagnostic isotope compositions are present.     

High prevalence,low counts and uncommon serotypes of Listeria monocytogenes in linguiça, a Brazilian fresh pork sausage

Linguiça is a highly popular and appreciated pork product in Brazil, frequently consumed undercooked. Aiming at collection of data for a future risk assessment, this study evaluated the prevalence and counts of Listeria monocytogenes in linguiça samples collected at retail level in Sao Paulo, SP, Brazil. ISO methods were used for detection and enumeration of the pathogen (11290-1 and 11290-2, respectively). Isolates were submitted to Simplex-PCR for hlyA gene and those with biochemical features of L. monocytogenes and hlyA positive were serotyped using a Multiplex PCR. Ninety percent of the samples were positive for Listeria spp., and L. monocytogenes was detected in 42% of the samples, with counts below 10² CFU/g in all samples. A prevalence of uncommon serotypes 4a and 4c was observed.     

Surveillance study of a number of synthetic and natural growth promoters in bovine muscle samples using liquid chromatography-tandem mass spectrometry

A simple, new method permitting the simultaneous determination and confirmation of trace residues of 24 different growth promoters and metabolites using liquid chromatography-mass spectrometry was developed and validated. The compounds were extracted from bovine tissue using acetonitrile; sodium sulphate was also added at this stage to aid with purification. The resulting mixture was then evaporated to approximately 1?ml and subsequently centrifuged at high speed and an aliquot injected onto the LC-MS/MS system. The calculated CCα values ranged between 0.11 and 0.46?µg?kg^?1; calculated CCβ were in the range 0.19–0.79?µg?kg^?1. Accuracy, measurement of uncertainty, repeatability and linearity were also determined for each analyte. The analytical method was applied to a number of bovine tissue samples imported into Ireland from third countries. Levels of progesterone were found in a number of samples at concentrations ranging between 0.28 and 30.30?µg?kg^?1. Levels of alpha- and beta-testosterone were also found in a number of samples at concentrations ranging between 0.22 and 8.63?µg?kg^?1 and between 0.16 and 2.08?µg?kg^?1 respectively.     

Determination of pyrethroids in chicken egg samples: development and validation of a confirmatory analytical method by gas chromatography/mass spectrometry

The aim of this study was to develop and validate an efficient analytical method based on gas chromatography/tandem mass spectrometry (GC/MS/MS) for detection and quantification of six pyrethroids residues (Phenothrin, Permethrin, Cyfluthrin, Cypermethrin, Deltamethrin and Fenvalerate) in chicken eggs. The method was based on a preliminary liquid–liquid extraction of albumen‐free yolk samples, followed by a clean‐up by solid‐phase extraction. GC/MS/MS analyses were carried out in the selected reaction monitoring mode. Validation parameters such as specificity, detection capability, decision limit, precision, recovery, stability and ruggedness were determined, resulting in compliance with Decision 2002/657/EC. No complicated apparatus are required; moreover, low volumes of organic solvents and a nonintensive manual labour are required. These low costs and simple procedure, based on rapid and safe operations, may represent a useful tool in the routine analysis of pyrethroids pesticides, in the place of the currently used conventional techniques.     

Quantification of organophosphate insecticides and herbicides in vegetable samples using the “Quick Easy Cheap Effective Rugged and Safe” (QuEChERS) method and a high-performance liquid chromatography-electrospray ionisation-mass spectrometry (LC-MS/MS) technique

A Quick Easy Cheap Effective Rugged and Safe (QuEchERS) extraction was developed and followed by selective analysis using a liquid chromatography-mass spectrometry method for the quantification of eighteen pesticides in vegetable samples. This method was accurate (?99.5%), and it exhibited limits of detection and quantification values in the 0.006-0.091 and 0.020-0.314 μg kg⁻¹ ranges, respectively. Furthermore, the coefficients of variations (?0.9999) were less than 1% at the low μg kg⁻¹ end of the method. Mean recoveries ranged between 94% and 102%, and relative standard deviations were below 10%. Based on these results, the methodology was proven to be highly efficient, robust, and suitable for monitoring the maximum residue limits (MRL) compliance of a wide range of commodity/pesticide combination. This method was successfully applied to the analysis of vegetable samples that were collected from different government farmers’ markets and street shops in urban areas. The presence of target pesticides was found in the range of 0.016 and 50.85 μg kg⁻¹.     

Direct characterisation of phenolic antioxidants in infusions from four Mapuche medicinal plants by liquid chromatography with diode array detection (HPLC-DAD) and electrospray ionisation tandem mass spectrometry (HPLC-ESI-MS)

A simple, fast and direct method was developed for the qualitative analysis of phenolic constituents from infusions of Mapuche medicinal plants. Teas made of Linum chamissonis Schiede, Quinchamalium chilensis Mol., Adesmia emarginata Clos. and Escallonia illinita K. Presl. were analysed by high-performance liquid chromatography with diode array detector (HPLC-DAD) and electrospray mass spectrometry (ESI-MS). This technique allowed for the first time the tentative identification of 16 phenolic compounds in E. illinita, 27 in Q. chilensis, 10 in L. chamissonis and 19 in A. emarginata. The compounds were mainly phenolic acids, flavonoid glycosides, anthocyanins and tannins. The total phenolic and flavonoid content of the infusions was assessed as well as the free radical scavenging capacity measured by the bleaching of a solution of the 2,2-diphenyl-1-picrylhydrazyl (DPPH⁻) radical. From the four species, Q. chilensis exhibited the strongest antioxidant activity with highest total phenolic and flavonoid content.     

Single cell-based analysis of torenia petal pigments by a combination of ArF excimer laser micro sampling and nano-high performance liquid chromatography (HPLC)-mass spectrometry

The molecular constituents of the petal pigments of the Torenia plant (Torenia hybrida) were analyzed on a single-cell basis by a combination of newly developed laser-microsampling and nano-flow liquid chromatography-electro spray ionization mass spectrometry (LC-ESIMS) techniques. Our method should provide a facile method for obtaining precise metabolic profiles of each cell in a single plant tissue.