Comparison of Ion Chromatography Conductivity Detection (IC-CD) and Ion Chromatography Inductively Coupled Plasma Mass Spectrometry (IC-ICP-MS) for the Determination of Phosphonic Acid in Grapevine Plant Parts, Wine, and Soil.

Potassium dihydrogen phosphonate had been allowed as a plant strengthener in organic viticulture in the European Union only until 2013, supporting the control of grapevine downy mildew. Therefore, low or nondetectable levels are a prerequisite for marketing of organic wines and, consequently, validated analytical methods are of major interest. Herein, two methods based on ion chromatography conductivity detection (IC-CD) or ion chromatography inductively coupled plasma mass spectrometry (IC-ICP-MS) for the determination of phosphonic acid (H3PO3) from 14 different plant matrices of Vitis vinifera L., wine, and soil were developed, validated, and compared. Extraction recoveries ranged from 95.1 to 99.3%. Limits of quantification (LOQ) ranged in liquid and solid samples from 3.8 to 16.8 µg/kg and 0.08 to 2.41 mg/kg for ICP-MS detection and from 39.9 to 593.7 µg/kg and 3.51 to 58.7 mg/kg for CD, respectively. Data on a current anonymized selection of 100 conventionally and 30 organically produced wines are briefly presented to demonstrate the suitability of the method.

1H NMR spectrometry for methanol quantification in apple wines and ciders as optimised by comparison to SIDA-HS-GC-MS.

Methanol, a hepato- and neurotoxic compound, is present in apple-based beverages as a by-product of the enzymatic degradation of pectin. Stable isotope dilution assay (SIDA) employing gas chromatography-mass spectrometry (GC-MS) is the state-of-the-art method for methanol determination in beverages. Despite higher initial investment costs, quantitative 1H NMR spectroscopy (qNMR) is a simpler and faster analytical technique to quantify numerous analytes in liquid foods. Beyond targeted analyses, qNMR spectral fingerprints in the product might be used for non-targeted analytical goals, such as adulteration and contamination detections. Here, an existing 1H-qNMR method used for wine profiling was optimised for methanol quantification in apple-based products, including cross-validation against a SIDA-HS-GC-MS method and reference values from interlaboratory trials. The optimisation involved a pivotally important estimation of a correction factor by an external calibration approach, making qNMR results comparable to SIDA-HS-GC-MS. The optimised qNMR method is suggested to be an alternative for methanol quantification in beverages.

Influence of processing steps on the fate of ochratoxin A, patulin, and alternariol during production of cloudy and clear apple juices.

Mycotoxins are frequently found in fruits and fruit juices. However, data about occurrence and fate of mycotoxins along the fruit juice processing chain are currently insufficient. Herein, a liquid chromatographic/tandem mass spectrometric (LC-MS/MS) multi-mycotoxin method was developed and applied to investigate the effect of technological unit operations on the fate of three of the most relevant mycotoxins along the processing chain for cloudy and clear apple juice, namely patulin (PAT), ochratoxin A (OTA), and alternariol (AOH). Raw juice obtained directly after dejuicing was spiked with the aforementioned mycotoxins at pilot-plant scale prior to subjecting it to different technological unit operations. Regarding clear apple juice production treatment with a pectinolytic enzyme preparation, and pasteurization were insignificant for mycotoxin reduction, but fining with subsequent filtration was effective, although the mycotoxins showed different affinity towards the tested agents. The most effective fining agent was activated charcoal/bentonite in combination with ultrafiltration, which removed OTA (54 µg/L) and AOH (79 µg/L) to not quantifiable amounts (limit of quantification (LOQ) 1.4 and 4.6 µg/L, respectively), while PAT was reduced only by 20% (from 396 to 318 µg/L). Regarding cloudy apple juice production, all studied processing steps such as centrifugation and pasteurization were ineffective in reducing mycotoxin levels. In brief, none of the common steps of clear and cloudy apple juice production represented a fully effective safety step for minimizing or even eliminating common mycotoxins. Thus, ensuring the sole use of sound apples should be of utmost importance for processors, particularly for those manufacturing cloudy juices.

Filter aid selection allows modulating the vanadium concentration in beverages.

This study reports a clear-cut relationship of the technological use of specific filter aids with highly variable vanadium levels in beer, wine, and fruit juices. First, the previously reported broad range of vanadium levels was confirmed in 68 commercial beverages by ICP-MS. Since cloudy apple juices exhibited significantly lower vanadium amounts than clear apple juices, filter aids used for clarification were analyzed and found to contain substantial and highly variable amounts of vanadium, particularly in all analyzed diatomite (38-368 mg vanadium per kg filter aid), but not in perlite products (<4 mg/kg). Subsequent pilot-scale precoat filtration experiments (170 L/batch) proved vanadium to be released from diatomite (Kieselguhr), increasing its levels from ca. 2.1-2.6 µg/kg unfiltered to 27-201 µg/kg filtered juice, depending on the use of diatomites high or low in vanadium. Thus, filter aid selection was shown to modulate the vanadium concentrations in clarified beverages.

Methylerythritol and mevalonate pathway contributions to biosynthesis of mono-, sesqui-, and diterpenes in glandular trichomes and leaves of Stevia rebaudiana Bertoni.

The biosynthesis of the diterpenoid steviol glycosides rebaudioside A and stevioside in nonrooted cuttings of Stevia rebaudiana was investigated by feeding experiments using the labeled key precursors [5,5-(2)H2]-mevalonic acid lactone (d2-MVL) and [5,5-(2)H2]-1-deoxy-d-xylulose (d2-DOX). Labeled glycosides were extracted from the leaves and stems and were directly analyzed by LC-(-ESI)-MS/MS and by GC-MS after hydrolysis and derivatization of the resulting isosteviol to the corresponding TMS-ester. Additionally, the incorporation of the proffered d2-MVL and d2-DOX into volatile monoterpenes, sesquiterpenes, and diterpenes in glandular trichomes on leaves and stems was investigated by headspace-solid phase microextraction-GC-MS (HS-SPME-GC-MS). Incorporation of the labeled precursors indicated that diterpenes in leaves and monoterpenes and diterpenes in glandular trichomes are predominately biosynthesized via the methylerythritol phosphate (MEP) pathway, whereas both the MEP and mevalonate (MVA) pathways contribute to the biosynthesis of sesquiterpenes at equal rates in glandular trichomes. These findings give evidence for a transport of MEP pathway derived farnesyl diphosphate precursors from plastids to the cytosol. Contrarily, the transport of MVA pathway derived geranyl diphosphate and geranylgeranyl diphosphate precursors from the cytosol to the plastid is limited.

Biosynthesis of sesquiterpenes in grape berry exocarp of Vitis vinifera L.: evidence for a transport of farnesyl diphosphate precursors from plastids to the cytosol.

The participation of the mevalonic acid (MVA) and 1-deoxy-d-xylulose 5-phosphate/2-C-methyl-d-erythritol-4-phosphate (DOXP/MEP) pathways in sesquiterpene biosynthesis of grape berries was investigated. There is an increasing interest in this class of terpenoids, since the oxygenated sesquiterpene rotundone was identified as the peppery aroma impact compound in Australian Shiraz wines. To investigate precursor supply pathway utilization, in vivo feeding experiments were performed with the deuterium labeled, pathway specific, precursors [5,5-(2)H2]-1-deoxy-d-xylulose and [5,5-(2)H2]-mevalonic acid lactone. Head Space-Solid Phase Micro Extraction-Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) analysis of the generated volatile metabolites demonstrated that de novo sesquiterpene biosynthesis is mainly located in the grape berry exocarp (skin), with no detectable activity in the mesocarp (flesh) of the Lemberger variety. Interestingly, precursors from both the (primarily) cytosolic MVA and plastidial DOXP/MEP pathways were incorporated into grape sesquiterpenes in the varieties Lemberger, Gewürztraminer and Syrah. Our labeling data provide evidence for a homogenous, cytosolic pool of precursors for sesquiterpene biosynthesis, indicating that a transport of precursors occurs mostly from plastids to the cytosol. The labeling patterns of the sesquiterpene germacrene D were in agreement with a cyclization mechanism analogous to that of a previously cloned enantioselective (R)-germacrene D synthase from Solidago canadensis. This observation was subsequently confirmed by enantioselective GC-MS analysis demonstrating the exclusive presence of (R)-germacrene D, and not the (S)-enantiomer, in grape berries.