Erica Liberto

Quantitative analysis of volatiles from solid matrices of vegetable origin by high concentration capacity headspace techniques: determination of furan in roasted coffee.

The study compares standard addition (SA), stable isotope dilution assay (SIDA) and multiple headspace extraction (MHE) as methods to quantify furan and 2-methyl-furan in roasted coffee with HS-SPME-GC-MS, using CAR-PDMS as fibre coating, d(4)-furan as internal standard and in-fibre internal standardization with n-undecane to check the fibre reliability. The results on about 150 samples calculated with the three quantitation approaches were all very satisfactory, with coefficient of variation (CV) versus the U.S. Food and Drug Administration (FDA) method, taken as reference, almost always below the arbitrarily-fixed limit of 15%. Furan was detected in the 1-5 ppm range, 2-methyl-furan in the 4-20 ppm range. Moreover, experimental exponential slopes (Q) and linearity (r) of both furan and 2-methyl-furan MHE regression equation on 50 samples were very similar thus making possible to use the same average Q value for all samples of the investigated set and their quantitation with a single determination. This makes this approach very rapid and competitive in-time with SA and SIDA. A non-separative method (HS-SPME-MS) was also developed in view of possible application on-line monitoring of furan and 2-methyl-furan in a pilot-plant with the aim of optimizing the roasting process to reduce these compounds to a minimum. Sampling times of 20 and 5 min were tested, the latter enabling total analysis time to be reduced to about 9 min. The results on 105 samples with both SIDA and MHE approaches were again highly satisfactory most of the samples giving a CV% versus the conventional methods below 20%. In this case too average Q values for both furan and 2-methyl-furan were used for MHE. The separative method presented very good repeatability (RSD% always below 10%) and intermediate precision over three months (RSD% always below 15%); performance were similar for the non-separative method, with repeatability (RSD%) always below 12% and intermediate precision over three months (RSD%) always below 15%. The sensitivity of both separative and non-separative methods was also very good, LOD and LOQ being in the ppb range for both furan and 2-methyl-furan, i.e. well below the amounts present in the roasted coffee samples.

New medium-to-high polarity twister coatings for liquid and vapour phase sorptive extraction of matrices of vegetable origin☆

Stir Bar Sorptive Extraction (SBSE) is a solventless sampling technique first introduced to extract organic analytes from aqueous samples, in the following applied to headspace sampling (HeadSpace Sorptive Extraction - HSSE). In SBSE and HSSE, analytes are sorbed onto a thick film of polydimethylsiloxane (PDMS) coating a glass-coated magnet. However, PDMS is apolar and not highly effective in recovering polar analytes (i.e. with logK(o/w) below 2), making difficult their sampling in complex matrices. A new generation of medium-to-high polarity polymeric coatings for twisters i.e. polyethyleneglycol-modified silicone (EG) and polyacrylate/polyethyleneglycol (PA) has recently been introduced to overcome this limit. In this study, EG and PA twisters have been applied to SBSE and HSSE of a number of dedicated standard mixtures and real-world samples of vegetable origin to evaluate their capability to increase recovery of medium to highly polar analytes. The results obtained, expressed as percent concentration factor (CF%) versus PDMS twisters taken as reference, showed that analyte logK(o/w) is a key-factor driving the choice of the most effective coating. The new twisters showed to be successful for both SBSE and HSSE, although to a different extent. EG twisters gave high recoveries with analytes over a wide range of polarities and in particular with logK(o/w) below 2 and/or containing hydroxyl or carboxylic functional groups independently on their logK(o/w). On the other hand, PA twisters were selectively effective for highly polar compounds with logK(o/w) below 1.

Quantitative determination of some volatile suspected allergens in cosmetic creams spread on skin by direct contact sorptive tape extraction–gas chromatography–mass spectrometry

This study describes a method based on direct contact sorptive tape extraction followed by on-line thermal desorption gas chromatography-mass spectrometry (DC-STE-GC-MS) to detect and quantify a group of suspected volatile allergens on the European Union (E.U.) list and a related compound on the skin (the stratum corneum) of volunteers treated with a cream of known composition fortified with the reference allergens. The following compounds were tested: citronellol, Z-citral (neral), geraniol, cinnamaldehyde, anisyl alcohol, cinnamyl alcohol, eugenol, methyleugenol, coumarin, isoeugenol, alpha-isomethylionone, 2-(4-tert-butylbenzyl)propionaldehyde (lilial), alpha-amylcinnamaldehyde, alpha-hexylcinnamaldehyde. Sorptive tape extraction (STE) is a sorption-based sampling technique in which a flexible polydimethylsiloxane (PDMS) tape is used to recover analytes by direct contact with the surface of a solid matrix or from the headspace in equilibrium with it. The reliability of the method was confirmed by: (i) allergen recoveries varying from 52.3% for lilial to 95.7% for neral, (ii) linearity in the range 10-150ppm, with regression coefficient R(2) always above 0.97, (iii) repeatability of each analyte, RSD% never exceeding 10%, (iv) intermediate precision, always below 15%, and (v) LOD and LOQ in the ppb range, therefore fully compatible with E.U. prescriptions (ppm). Other parameters such as substantivity analyte, approximate permeation through skin and influence of different nature of stratum corneum on recovery were also investigated. The method was also successfully applied to five commercially available creams declared to contain some of the allergens in question spread on the skin of the same volunteers.

Conventional and narrow bore short capillary columns with cyclodextrin derivatives as chiral selectors to speed-up enantioselective gas chromatography and enantioselective gas chromatography–mass spectrometry analyses

The analysis of complex real-world samples of vegetable origin requires rapid and accurate routine methods, enabling laboratories to increase sample throughput and productivity while reducing analysis costs. This study examines shortening enantioselective-GC (ES-GC) analysis time following the approaches used in fast GC. ES-GC separations are due to a weak enantiomer-CD host-guest interaction and the separation is thermodynamically driven and strongly influenced by temperature. As a consequence, fast temperature rates can interfere with enantiomeric discrimination; thus the use of short and/or narrow bore columns is a possible approach to speeding-up ES-GC analyses. The performance of ES-GC with a conventional inner diameter (I.D.) column (25 m length x 0.25 mm I.D., 0.15 microm and 0.25 microm d(f)) coated with 30% of 2,3-di-O-ethyl-6-O-tert-butyldimethylsilyl-beta-cyclodextrin in PS-086 is compared to those of conventional I.D. short column (5m length x 0.25 mm I.D., 0.15 microm d(f)) and of different length narrow bore columns (1, 2, 5 and 10 m long x 0.10 mm I.D., 0.10 microm d(f)) in analysing racemate standards of pesticides and in the flavour and fragrance field and real-world-samples. Short conventional I.D. columns gave shorter analysis time and comparable or lower resolutions with the racemate standards, depending mainly on analyte volatility. Narrow-bore columns were tested under different analysis conditions; they provided shorter analysis time and resolutions comparable to those of conventional I.D. ES columns. The narrow-bore columns offering the most effective compromise between separation efficiency and analysis time are the 5 and 2m columns; in combination with mass spectrometry as detector, applied to lavender and bergamot essential oil analyses, these reduced analysis time by a factor of at least three while separation of chiral markers remained unaltered.

Room temperature ionic liquids: New GC stationary phases with a novel selectivity for flavor and fragrance analyses ☆

Ionic liquids (ILs) are of great interest as moderately polar to polar stationary phases for GC, because their selectivity differs markedly from that of conventionally used phases. In the flavor, fragrance and essential oil fields, analysts often deal with complex mixtures of compounds having similar structural and physical characteristics (e.g., mono- and sesquiterpenoids), therefore requiring an interactive combination between chromatographic and mass spectral data for correct identification. New GC stationary phases with different selectivity must therefore be continually tested. Performance and evolution over time of commercially available IL columns versus those commonly used in these fields are here evaluated, mainly in view of their routine use. Chromatographic and separative properties (efficiency, separation capability, inertness and/or activity) of commercially available IL columns were compared to those of columns coated with 5% phenyl-95% methylpolysiloxane, 14% cyanopropyl-86% polysiloxane, and polyethylene glycol, on different complexity samples, including standard mixtures of volatile suspected allergens and pesticides, and cornmint and vetiver essential oils. The results show that IL columns can successfully be used for a wide range of applications characteristic of these fields, mainly because of their unusual selectivity, in particular when separations based on functional groups are required. Moreover, the latest generation of IL columns (IL61 and IL60) presents chromatographic performance comparable to or only slightly lower than that of the conventional columns routinely used in these fields.

Development of fast enantioselective gas-chromatographic analysis using gas-chromatographic method-translation software in routine essential oil analysis (lavender essential oil).

The study aimed to find the best trade-off between separation of the most critical peak pair and analysis time, in enantioselective GC-FID and GC-MS analysis of lavender essential oil, using the GC method-translation approach. Analysis conditions were first optimized for conventional 25 m x 0.25 mm inner diameter (dc) column coated with 6(I-VII)-O-tert-butyldimethylsilyl-2(I-VII)-3(I-VII)-O-ethyl-beta-cyclodextrin (CD) as chiral stationary phase (CSP) diluted at 30% in PS086 (polymethylphenylpolysiloxane, 15% phenyl), starting from routine analysis. The optimal multi-rate temperature program for a pre-set column pressure was determined and then used to find the pressures producing the efficiency-optimized flow (EOF) and speed-optimized flow (SOF). This method was transferred to a shorter narrow-bore (NB) column (11 m x 0.10 mm) using method-translation software, keeping peak elution order and separation. Optimization of the enantioselective GC method with the translation approach markedly reduced the analysis time of the lavender essential oil, from about 87 min with the routine method to 40 min with an optimal multi-rate temperature program and initial flow with a conventional inner diameter column, and to 15 min with FID as detector or 13.5 min with MS with a corresponding narrow-bore column, while keeping enantiomer separation and efficiency.

High-quality Italian rice cultivars: Chemical indices of ageing and aroma quality

The volatile fractions of six Italian high-quality rice cultivars were investigated by HS-SPME-GC-MS to define fingerprinting and identify chemical markers and/or indices of ageing and aroma quality. In particular, four non-aromatic (Carnaroli, Carnise, Cerere and Antares) and two aromatic (Apollo and Venere) rices, harvested in 2010 and 2011, were monitored over 12months. Twenty-five aroma components were considered and, despite considerable inter-annual variability, some of them showed similar trends over time, including 2-(E)-octenal as a marker of ageing for all cultivars, and heptanal, octanal and 2-ethyl hexanol as cultivar-specific indicators. The area ratios 2-acetyl-1-pyrroline/1-octen-3-ol, for Venere, and 3-methyl-1-butanol/2-methyl-1-butanol, for Apollo, were also found to act as ageing indices. Additional information on release of key-aroma compounds was also obtained from quantitation and its dependence on grain shape and chemical composition. Heptanal/1-octen-3-ol and heptanal/octanal ratios were also defined as characterising the aroma quality indices of the six Italian rice cultivars investigated.

New asymmetrical per-substituted cyclodextrins (2-O-methyl-3-O-ethyl- and 2-O-ethyl-3-O-methyl-6-O-t-butyldimethylsilyl-β-derivatives) as chiral selectors for enantioselective gas chromatography in the flavour and fragrance field

Asymmetrically substituted 6(I-VII)-O-t-butyldimethylsilyl(TBDMS)-3(I-VII)-O-ethyl-2(I-VII)-O-methyl-beta-cyclodextrin (MeEt-CD) and 6(I-VII)-O-TBDMS-2(I-VII)-O-ethyl-3(I-VII)-O-methyl-beta-cyclodextrin (EtMe-CD) were synthesised to evaluate the role of the substitution pattern in positions 2 and 3 on the enantioselectivity, in particular in view of their application to routine analysis in fast enantioselective gas chromatography (Es-GC). The chromatographic properties and enantioselectivities of the new derivatives were tested by separating the enantiomers of a series of medium-to-high volatility racemates in the flavour and fragrance field, and compared to those of the corresponding symmetrically substituted 6(I-VII)-O-TBDMS-2(I-VII),3(I-VII)-O-methyl-beta-CD (MeMe-CD) and 6(I-VII)-O-TBDMS-2(I-VII),3(I-VII)-O-ethyl-beta-CD (EtEt-CD), and were then applied to analysis of real-world essential oil (e.o.) samples. A new synthetic process including the sonochemical approach to obtain synthetic reproducibility and significant yields of the per-substituted derivatives with acceptable reaction times was developed. The results show that asymmetrically substituted methyl/ethyl CDs compared to the methyl or ethyl symmetrical derivatives in general provide better enantioselectivity in terms of both enantiomer resolution and number of separated chiral compounds, and show how the substitution pattern in positions 2 and 3 of the CD ring can influence the separation. Moreover, these new CD derivatives with better enantioselectivity are also shown to be very useful in routine analysis for the exhaustive control of samples containing several chiral characterizing markers in a single run.

Volatile profiling of high quality hazelnuts (Corylus avellana L.): chemical indices of roasting.

The study proposes an investigation strategy to identify sensitive, robust and reliable chemical markers of hazelnut roasting. A fully-automated and validated analytical method, based on Headspace Solid Phase Microextraction (HS-SPME) coupled with Gas Chromatography-Mass Spectrometric detection (GC-MS), for effective off-line monitoring of changes in the volatile profile of high-quality hazelnuts was developed. Samples from two different harvests were submitted to roasting, following different time/temperature protocols and different technologies, enabling chemical changes to be correlated with technological processing and sensory quality. Chemical indices, expressed as analyte response ratio, were defined and their trend observed across roasting profiles. Reliability and robustness of chemical indices were also evaluated, in view of their application to on-line monitoring with Mass Spectrometry-based electronic nose technology (MS-nose). Experiments, simulating on-line chemical characterisation of the volatile fraction, were performed through a fully-automated system. The results confirmed: (a) the effectiveness of single process indicators of roasting selected by the separative method (5-methylfurfural, 1(H)-pyrrole, furfuryl alcohol, 1(H)-pyrrole-2-carboxaldehyde, 1-hydroxy-2-propanone, dihydro-2(3H)-furanone, 5-methyl-(E)-2-hepten-4-one, acetic acid, pyridine, furfural, pyrazine, and several alkyl-pyrazines); and, (b) the reliability of proposed chemical indices: 5-methylfurfural/2,5-dimethylpyrazine, 5-methylfurfural/2-methylpyrazine, 2,5-dimethylpyrazine/2,3-dimethylpyrazine; these maintained a consistent trend versus harvest and sampling/analysis technology.

Urinary metabolic fingerprinting of mice with diet-induced metabolic derangements by parallel dual secondary column-dual detection two-dimensional comprehensive gas chromatography

This study investigates the potential of a parallel dual secondary column-dual detection two-dimensional comprehensive GC platform (GC×2GC-MS/FID) for metabolic profiling and fingerprinting of mouse urine. Samples were obtained from a murine model that mimics a typical unhealthy Western diet featuring both high fat and sugar (HFHS) intake, which induces obesity, dyslipidemia, and insulin resistance. Urines collected at different steps of the study were used to obtain pivotal and comparative data on the presence and relative distributions of early markers of metabolic disease. The data elaboration and interpretation work-flow includes an advanced untargeted fingerprinting approach, with peak-region features to locate relevant features to be quantified by external standard calibration. The reliability of untargeted fingerprinting is confirmed by quantitative results on selected relevant features that showed percentages of variations consistent with those observed by comparing raw data quantitative descriptors (2D peak-region volumes and percent of response). Analytes that were up-regulated with % of variation ranging from 30 to 1000, included pyruvic acid, glycerol, fructose, galactose, glucose, lactic acid, mannitol and valine. Down-regulation was evidenced for malonic acid, succinic acid, alanine, glycine, and creatinine. Advanced fingerprinting also is demonstrated for effectively evaluating individual variations during experiments, thus representing a promising tool for personalized intervention studies. In this context, it is interesting to observe that informative features that were not discriminant for the entire population may be relevant for individuals.