Hans-Georg Schmarr

Coupled liquid chromatography-gas chromatography for the rapid analysis of γ-oryzanol in rice lipids

An approach based on on-line coupled liquid chromatography-gas chromatography (LC-GC) was developed for the rapid analysis of gamma-oryzanol in rice. Total lipids were extracted from rice and subjected to LC-GC without any prior purification. gamma-Oryzanol was pre-separated by HPLC from rice lipids and transferred on-line to GC analysis in order to separate its major constituents. 24-methylenecycloartanyl ferulate, cycloartenyl ferulate, campesteryl ferulate, beta-sitosteryl ferulate and campestanyl ferulate. The identities of the compounds were confirmed by off-line GC-MS analysis. Total gamma-oryzanol content could be quantified by HPLC-UV detection and the distribution of gamma-oryzanol constituents could be determined by on-line coupled GC analysis. The proposed methodology paves the way for high-throughput investigations providing information on natural variations in gamma-oryzanol content and its composition in different rice varieties.

Analysis of steryl esters in cocoa butter by on-line liquid chromatography–gas chromatography

On-line liquid chromatography-gas chromatography (LC-GC) has been applied to the analysis of steryl esters in cocoa butter. Separation of the steryl esters was achieved after on-line transfer to capillary GC. HPLC removes the large amount of triglycerides and pre-separates the components of interest, thus avoiding time-consuming sample preparation prior to GC analysis. The identities of the compounds were confirmed by GC-MS investigation of the collected HPLC fraction and by comparison of the mass spectra (chemical ionization using ammonia as ionization gas) to those of synthesized reference compounds. Using cholesteryl laurate as internal standard, steryl esters were quantified in commercial cocoa butter samples, the detection limit being 3 mg/kg and the quantification limit 10 mg/kg, respectively. Only slight differences in percentage distributions of steryl esters depending on the geographical origin of the material were observed. The patterns were shown to remain unchanged after deodorization. The method described might be a valuable tool for authenticity assessment of cocoa butter.

Analysis of carbonyl compounds via headspace solid-phase microextraction with on-fiber derivatization and gas chromatographic–ion trap tandem mass spectrometric determination of their O-(2,3,4,5,6-pentafluorobenzyl)oxime derivatives

An improved method for the analysis of carbonyls is described utilizing a headspace solid-phase microextraction (HS-SPME) step and on-fiber derivatization with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) hydrochloride. Thermal desorption of the oxime derivatives formed on the fiber is followed by gas chromatographic separation coupled to an ion trap tandem mass spectrometer (GC-ITMS). Selecting specific fragment ions within the electron ionization (EI(+)) mass spectra of these oxime derivatives as precursor ions for MS-MS fragmentation provides a suitable method for the target analysis of individual carbonyl classes, such as alkanals, (E)-2-alkenals, (E,E)-2,4-alkadienals, and others. Retention indices on polar as well as on apolar stationary phases along with EI(+) mass spectra patterns are presented for a large set of oxime derivatives, giving valuable information needed for unambiguous assignment of substances in complex sample matrices. The fast sample preparation and derivatization step via HS-SPME can be automated and is applicable to a variety of biological samples and foodstuffs, allowing rapid and sensitive screening analyses of important aldehydic biomarkers and aroma active compounds.

Pitfalls encountered during quantitative determination of 3-alkyl-2-methoxypyrazines in grape must and wine using gas chromatography–mass spectrometry with stable isotope dilution analysis. Comprehensive two-dimensional gas chromatography–mass spectrometry and on-line liquid chromatography-multidimensional gas chromatography–mass spectrometry as potential loopholes

The analysis of 3-alkyl-2-methoxypyrazines in Vitis vinifera grape must or wine at the low nanogram per liter level failed in several situations when applying a one-dimensional gas chromatographic analysis with mass spectrometric detection (GC-MS). Sample preparation methods such as headspace solid phase microextraction or solid phase extraction were convenient procedures, however lacking extraction selectivity for complex matrices. Analysis by comprehensive two-dimensional gas chromatography with mass spectrometric detection clearly demonstrated the potential for co-elution in such matrices and the risk for erroneous results when applying one-dimensional GC-MS. In one example, matrix problems would have been a challenge even for a comprehensive two-dimensional chromatographic approach with MS detection (GC×GC-MS). A solution to matrix problems was found by protonating the 3-alkyl-2-methoxypyrazines in acidic pH and sample clean-up using solid phase extraction with a mixed-mode polymeric cation-exchange sorbent. Quantification was performed by a stable isotope dilution assay, following analysis by on-line coupled high performance liquid chromatography with multidimensional gas chromatography and detection with mass spectrometry (on-line LC-MDGC-MS). This new approach allowed trace-level analysis of 3-alkyl-2-methoxypyrazines in grape musts and wines and is described for V. vinifera Sauvignon blanc, following 3-alkyl-2-methoxypyrazines concentrations during ripening and in the processed wines.

Analysis of aldehydes via headspace SPME with on-fiber derivatization to their O-(2,3,4,5,6-pentafluorobenzyl)oxime derivatives and comprehensive 2D-GC-MS

A method for the analysis of the homologous series of alkanals, (E)-2-alkenals, and (E,E)-2,4-alkadienals is described utilizing a headspace solid-phase microextraction (HS-SPME) step and on-fiber derivatization with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) hydrochloride. Oxime derivatives formed on the fiber are desorbed in the gas chromatographic injector and analyzed by comprehensive 2-D GC coupled to quadrupole MS (GC x GC-qMS). Selecting specific fragment ions within the electron impact mass spectra of the oxime derivatives provides a suitable method for the target analysis of these aldehyde classes, which furthermore benefits from the increased separation efficiency by GC x GC. The analysis of higher molecular weight aldehydes is described in wine and grape seed oil as examples. Quantification of the aldehydes utilizes a stable isotope dilution analysis (SIDA) assay with octan-d(16)-al as isotopomeric internal standard. Besides the selectivity and sensitivity of aldehyde analysis using PFBHA derivatives, critical aspects on background level contamination and repeatability of the sample preparation method are discussed. Optimization of GC x GC-qMS parameters allowed a considerable saving of the cryogenic medium, involving additional (unmodulated) conditioning runs, rendering the method more amenable to routine analysis.

Rapid detection of vegetable oils in milk fat by on-line LC-GC analysis of β-sitosterol as marker

On-line liquid chromatography-gas chromatography (LC-GC) has been applied to the detection of vegetable oils in milk fat using β-sitosterol as marker. The method involves transesterification of the fat, pre-separation of the sterol fraction from other lipid constituents and on-line transfer to the capillary GC system. The on-line approach avoids time-consuming sample preparation steps prior to GC analysis. The suitability of this analytical approach was tested with model mixtures of milk fat with cotton and rapeseed oil. The method allows the detection of adulterations at low levels. Simultaneous quantification of cholesterol in milk fat is also possible. Considering the rapid sample preparation, the described method can be applied for screening of large sample numbers.

Two-dimensional gas chromatographic profiling as a tool for a rapid screening of the changes in volatile composition occurring due to microoxygenation of red wines.

Microoxygenation (MOX) is a widely applied technique to deliver continuously trace amounts of oxygen to red wine during vinification and ageing in order to improve color stability and sensory properties. Proven by sensory means, the added oxygen modifies not only tannin structure and color of the wines, but also their composition of volatiles. In this study different microoxygenation treatments prior and after malolactic fermentation were carried out for Pinot noir, Cabernet Sauvignon and Dornfelder wines of the 2007 vintage. Volatile components of subsequent wines were analyzed using headspace-solid phase microextraction coupled to comprehensive two-dimensional gas chromatography-quadrupole mass spectrometry (GC x GC-qMS). Quantitative data were retrieved from two-dimensional images obtained from GC x GC chromatograms of volatile compounds applying a software package, which is commonly used in the field of proteomics for two-dimensional electrophoresis gels. This approach revealed a discrimination of the applied treatments by multivariate statistics based on volatiles alone, such as the clear distinction among wines treated before or after malolactic fermentation in case of Cabernet Sauvignon and Dornfelder or the effect of different oxygen doses. Besides the differentiation of MOX treatments from the untreated control, specific varietal and technological effects could be distinguished. The image processing of the GC x GC data offered valuable tools which were able to identify those areas in the 2D images that were most responsible for discrimination among different MOX treatments. Based on the loadings of individual aroma compounds a set of markers for the MOX-induced modifications of volatiles could be suggested.

Stereodifferentiation of some chiral monoterpenes using multidimensional gas chromatography

SummaryHeptakis (2,3,6-tri-O-alkyl)-β-cyclodextrins are used as chiral stationary phases for the resolution of α-pinene (1), β-pinene (2), limonene (3), α-terpineol (4), linalool (5) and the furanoid cis/trans linalool oxides (6) by chirospecific CGC-analysis. The first investigations on the direct chirality evaluation of the monoterpenoid grape and wine flavour compounds 4–6 are reported using enantioselective multidimensional gas chromatography (MDGC) and chiral main columns, coated with peralkylated β-cyclodextrins. The fresh grape juice, the corresponding must, the young and the bottle maturated wine of muscat cultivars are investigated.

Trace level analysis of corky off-flavor compounds: development of a new analytical method based on solid phase extraction and analysis by multidimensional gas chromatography with mass spectrometric detection.

This work describes the development of a trace level (<1 ngL(-1)) analysis of haloanisoles in complex wine matrix. The suggested method involves sample preparation based on solid phase extraction, a clean-up to remove acidic compounds, concentration of the haloanisole fraction and large volume on-column injection into a multidimensional GC-MS system. Mass spectrometric detection in the selected ion mode allowed reliable quantification of 2,4,6-trichloroanisole (TCA) or 2,4,6-tribromoanisole (TBA), via their highly deuterated ([(2)H5]) isotopologues as internal standards (stable isotope dilution analysis; SIDA), which had prior been synthesized in house. The development of this new method had become necessary, as a one-dimensional HS-SPME-GC-ECD method, routinely applied for analysis of TCA in cork soaks, had to be extended for TeCA and TBA determination, but failed due to co-elutions within wine matrices. The newly developed SPE//MDGC-MS method provided detection limits well below olfactory thresholds of the analytes with 0.05 ngL(-1) (LOD), 0.19 ngL(-1) (LOQ) for TCA, 0.06 ngL(-1) (LOD), 0.21 ngL(-1) (LOQ) for TeCA, and 0.09 ngL(-1) (LOD), 0.34 ngL(-1) (LOQ) for TBA.

Simultaneous stereoanalysis of 2-alkyl-branched acids, esters and alcohols using a selectivity-adjusted column system in multi-dimensional gas chromatography

Abstract The direct and simultaneous stereodifferentiation of 2-methylbutanoic acid, 2-methylbutanoic acid methyl and ethyl esters and the corresponding alcohol 2-methylbutane-1-ol from complex matrices is achieved, using selectivity-adjusted multi-dimensional gas chromatography with perethylated β-cyclodextrin as the chiral stationary phase.