The use of proton transfer reaction mass spectrometry for high throughput screening of terpene synthases.

In this work, we introduce the application of proton transfer reaction mass spectrometry (PTR-MS) for the selection of improved terpene synthase mutants. In comparison with gas chromatography mass spectrometry (GC-MS)-based methods, PTR-MS could offer advantages by reduction of sample preparation steps and analysis time. The method we propose here allows for minimal sample preparation and analysis time and provides a promising platform for the high throughput screening (HTS) of large enzyme mutant libraries. To investigate the feasibility of a PTR-MS-based screening method, we employed a small library of Callitropsis nootkatensis valencene synthase (CnVS) mutants. Bacterial cultures expressing enzyme mutants were subjected to different growth formats, and headspace terpenes concentrations measured by PTR-Qi-ToF-MS were compared with GC-MS, to rank the activity of the enzyme mutants. For all cultivation formats, including 96 deep well plates, PTR-Qi-ToF-MS resulted in the same ranking of the enzyme variants, compared with the canonical format using 100 mL flasks and GC-MS analysis. This study provides a first basis for the application of rapid PTR-Qi-ToF-MS detection, in combination with multi-well formats, in HTS screening methods for the selection of highly productive terpene synthases.

Portraying and tracing the impact of different production systems on the volatile organic compound composition of milk by PTR-(Quad)MS and PTR-(ToF)MS.

The aim of this study was to discover the unique volatile compositional traits of retail milk from different production systems. Forty-four retail milk samples were analyzed, including organic milk (n=10), conventional milk (n=14) and pasture milk (n=20) from winter (n=22) and summer (n=22). Proton transfer reaction quadrupole mass spectrometry (PTR-(Quad)MS) was utilized to obtain the mass-resolved fingerprints (76 masses per sample) of volatile organic compounds (VOCs). Principal component analysis (PCA) and analysis of variance (ANOVA) were performed to evaluate the differences between the groups. The production systems were characterized by six masses, while season showed larger differences, with twenty-two masses discriminating between the milks. For 2 masses, a significant interaction of systems and seasons was observed. The chemical formula of these VOC masses were tentatively identified by Proton Transfer Reaction Time-of-Flight Mass Spectrometric (PTR-(ToF)MS). These results illustrate that the type of feed is reflected in the VOC composition of milks.

Investigation of the aroma of commercial peach (Prunus persica L. Batsch) types by Proton Transfer Reaction-Mass Spectrometry (PTR-MS) and sensory analysis.

The aim of this study was to investigate the aroma and sensory profiles of various types of peaches (Prunus persica L. Batsch.). Forty-three commercial cultivars comprising peaches, flat peaches, nectarines, and canning peaches (pavías) were grown over two consecutive harvest years. Fruits were assessed for chemical aroma and sensory profiles. Chemical aroma profile was obtained by proton transfer reaction-mass spectrometry (PTR-MS) and spectral masses were tentatively identified with PTR-Time of Flight-MS (PTR-Tof-MS). Sensory analysis was performed at commercial maturity considering seven aroma/flavor attributes. The four types of peaches showed both distinct chemical aroma and sensory profiles. Flat peaches and canning peaches showed most distinct patterns according to discriminant analysis. The sensory data were related to the volatile compounds by partial least square regression. γ-Hexalactone, γ-octalactone, hotrienol, acetic acid and ethyl acetate correlated positively, and benzeneacetaldehyde, trimethylbenzene and acetaldehyde negatively to the intensities of aroma and ripe fruit sensory scores.

Proton-transfer reaction mass spectrometry (PTR-MS) for the authentication of regionally unique South African lamb.

The volatile fingerprints of South African lamb meat and fat were measured by proton-transfer mass spectrometry (PTR-MS) to evaluate it as an authentication tool. Meat and fat of the Longissimus lumborum (LL) of lambs from six different regions were assessed. Analysis showed that the volatile fingerprints were affected by the origin of the meat. The classification of the origin of the lamb was achieved by examining the calculated and recorded fingerprints in combination with chemometrics. Four different partial least squares discriminant analysis (PLS-DA) models were fitted to the data to classify lamb meat and fat samples into "region of origin" (six different regions) and "origin" (Karoo vs. Non-Karoo). The estimation models classified samples 100% correctly. Validation of the first two models gave 42% (fat) and 58% (meat) correct classification of region, while the second two models performed better with 92% (fat) and 83% (meat) correct classification of origin.

Characterization of Retail Conventional, Organic, and Grass Full-Fat Butters by Their Fat Contents, Free Fatty Acid Contents, and Triglyceride and Fatty Acid Profiling.

In the Netherlands, butter is produced from milk originating from three different production systems: conventional, organic, and grass-fed cows. The aim of the current study was to characterize these types of butters, and pinpoint distinct compositional differences. Retail conventional (n = 28), organic (n = 14), and grass (n = 12) full-fat butters were collected during the winter and summer seasons. Samples were analyzed for their fat content, free fatty acid (FFA) content, and triglyceride (TG) and fatty acid (FA) profiles. The fat content was significantly lower in conventional butters than in organic butters and the FFA content was significantly lower in conventional butters compared with grass butters. Also, organic butters differed significantly from their conventional counterparts with regard to their TG and FA profiles. The TG profiles of the organic and grass butters did not differ significantly. The FA profiles of grass butters were less distinct, since only a few FAs differed significantly from conventional (six FAs) and organic (eight FAs) butters.

Authentication of geographical origin and crop system of grape juices by phenolic compounds and antioxidant activity using chemometrics.

The main goal of this work was to propose an authentication model based on the phenolic composition and antioxidant and metal chelating capacities of purple grape juices produced in Brazil and Europe in order to assess their typicality. For this purpose, organic, conventional, and biodynamic grape juices produced in Brazil (n = 65) and in Europe (n = 31) were analyzed and different multivariate class-modeling and classification statistical techniques were employed to differentiate juices based on the geographical origin and crop system. Overall, Brazilian juices, regardless of the crop system adopted, presented higher contents of total phenolic compounds and flavonoids, total monomeric anthocyanins, proanthocyanidins, flavonols, flavanols, cyanidin-3-glucoside, delphinidin-3-glucoside, and malvidin-3,5-glucoside. No differences were observed for trans-resveratrol, malvidin-3-glucoside, and pelargonidin-3-glucoside between countries and among crop systems. A total of 91% of Brazilian and 97% of European juices were adroitly classified using partial least squares discriminant analysis when the producing region was considered (92% efficiency), in which the free-radical scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl, content of total phenolic compounds, gallic acid, and malvidin-3-glucoside were the variables responsible for the classification. Intraregional models based on soft independent modeling of class analogy were able to differentiate organic from conventional Brazilian juices as well as conventional and organic/biodynamic European juices.

Geographical provenancing of purple grape juices from different farming systems by proton transfer reaction mass spectrometry using supervised statistical techniques.

BACKGROUND: Organic, biodynamic and conventional purple grape juices (PGJ; n = 79) produced in Brazil and Europe were characterized by volatile organic compounds (m/z 20-160) measured by proton transfer reaction mass spectrometry (PTR-MS), and classification models were built using supervised statistical techniques. RESULTS: k-Nearest neighbours and soft independent modelling of class analogy (SIMCA) models discriminated adequately the Brazilian from European PGJ (overall efficiency of 81% and 87%, respectively). Partial least squares discriminant analysis (PLSDA) classified 100% European and 96% Brazilian PGJ. Similarly, when samples were grouped as either conventional or organic/biodynamic, the PLSDA model classified 81% conventional and 83% organic/biodynamic juices. Intraregional PLSDA models (juices produced in the same region - either Europe or Brazil) were developed and were deemed accurate in discriminating Brazilian organic from conventional PGJ (81% efficiency), as well as European conventional from organic/biodynamic PGJ (94% efficiency). CONCLUSIONS: PGJ from Brazil and Europe, as well as conventional and organic/biodynamic PGJ, were distinguished with high efficiency, but no statistical model was able to differentiate organic and biodynamic grape juices. These data support the hypothesis that no clear distinction between organic and biodynamic grape juices can be made with respect to volatile organic compounds.

Seafood and Water Management.

Seafood is an important food source for many. Consumers should be entitled to an informed choice, and there is growing concern about correct composition labeling of seafood. Due to its high price, it has been shown to be vulnerable to adulteration. In the present study, we focus on moisture levels in seafood. Moisture and crude protein contents of chilled and frozen cod, pangasius, salmon, shrimp and tilapia purchased from various retail outlets in the Netherlands were examined by reference methods and the values of which were compared with the reported data from other studies in literature. Significant differences in proximate composition were determined for different species and between chilled and frozen products of the same species. Pangasius products showed the highest moisture contents in general (86.3 g/100 g), and shrimp products revealed the largest differences between chilled and frozen products. Comparison with literature values and good manufacturing practice (GMP) standards exposed that, generally, chilled pangasius, frozen pangasius and frozen shrimp products presented considerably higher moisture and lower crude protein/nitrogen contents than those found in other studies. From the GMP standards, extraneous water was estimated on average at 26 g/100 g chilled pangasius product, and 25 and 34 g/100 g product for frozen shrimp and pangasius products, respectively.

Typicality and geographical origin markers of protected origin cheese from The Netherlands revealed by PTR-MS.

Volatile fingerprints of 30 cumin cheese samples of artisanal farmers' cheese of Leiden with EU Protected Designation of Origin (PDO) and 29 cumin cheese samples of varying commercial Dutch brands without PDO protection were used to develop authentication models. The headspace concentrations of the volatiles, as measured with high sensitivity proton-transfer mass spectrometry, were subsequently subjected to partial least-squares discriminant analysis (PLS-DA). Farmers' cheese of Leiden showed a distinct volatile profile with 27 and 9 out of the 60 predominant ions showing respectively significantly higher and lower concentrations in the headspace of the cheese in comparison to the other cumin cheeses. The PLS-DA prediction models developed classified in cross-validation 96% of the samples of PDO protected, artisanal farmers' cheese of Leiden correctly, against 100% of commercial cumin cheese samples. The characteristic volatile compounds were tentatively identified by PTR-time-of-flight-MS. A consumer test indicated differences in appreciation, overall flavor intensity, creaminess, and firmness between the two cheese groups. The consumers' appreciation of the cumin cheese tested was not influenced by the presence of a name label or PDO trademark.