Atmospheric pressure chemical ionization mass spectrometry of retro-carotenoids.

RATIONALE: The single and double bonds of the polyene chain of the studied retro-carotenoids are located at the neighboring positions compared to those of regular carotenoids. Our mass spectrometry approach targeted at facilitating the characterization of retro-carotenoids as their structural diversity in nature is not yet fully elucidated. Moreover, extended π-electron systems endow several retro-carotenoids with exceptional colors from golden-orange to vibrant red that stimulate the food industry's interest. METHODS: Atmospheric pressure chemical ionization-quadrupole time-of-flight-high-resolution mass spectrometry (APCI-QTOF-HRMS) experiments of the three structurally related retro-carotenoids rhodoxanthin, eschscholtzxanthone, and eschscholtzxanthin were performed to elucidate the formation of specific ion species compared to those of the common carotenoids lutein and zeaxanthin. Mass fragmentations of the aforementioned retro-carotenoids were unraveled using APCI-tandem mass spectrometry (MS/MS) in the negative and positive ion modes. RESULTS: Abundant in-source fragment ions [M + H - H2 O]+ of eschscholtzxanthin and eschscholtzxanthone were formed in the positive ion mode owing to the loss of water at the hydroxylated ε-rings. Eliminations of the ε-rings at the characteristic exocyclic double bonds at C-6,7 and C-6',7' were observed after the resonance-stabilized loss of water. Distinct product ions were yielded for all retro-carotenoids assessed because of the cleavage at their typical central single bond at C-15,15'. CONCLUSIONS: Detailed APCI-QTOF-HRMS analyses enabled a highly accurate detection of the most abundant ion species and respective signal intensity ratios of retro-carotenoids, facilitating their further screening and reliable identification in natural sources. Mass fragmentations of the studied retro-carotenoids were found to be substantially impacted by the extraordinary configuration of their polyene backbone.

Comparative Evaluation of Juices from Red-Fleshed Apples after Production with Different Dejuicing Systems and Subsequent Storage.

In this work, two vintages (2019 and 2020) of red-fleshed 'Weirouge' apples were processed with the innovative spiral filter press technology to investigate juice production in an oxygen-reduced atmosphere. After pressing, a more brilliant red color and appreciably higher amounts of oxidation-sensitive constituents (ascorbic acid, anthocyanins, and colorless (poly)phenols) were seen in spiral filter pressed juices compared to those produced with conventional systems (horizontal filter press and decanter). In a subsequent stability study (24 weeks storage at 4, 20, and 37 °C), the color and phenolic compounds were monitored and differences in the juices produced with the different pressing-systems were widely maintained during the storage period. The analyses of the anthocyanins and colorless (poly)phenols were conducted by UHPLC-DAD-ESI-QTOF-HR-MS/MS and UHPLC-DAD. The spiral filter press emerged as a promising technology for the production of juices with a more attractive color and a better retention of oxidation-sensitive constituents during processing and storage compared to conventional juices.

Studies into the phenolic patterns of different tissues of pineapple (Ananas comosus [L.] Merr.) infructescence by HPLC-DAD-ESI-MS (n) and GC-MS analysis.

In a comprehensive study, more than 60 phenolic compounds were detected in methanolic extracts from different tissues of pineapple infructescence by high-performance liquid chromatography with diode array detection and electrospray ionisation multiple-stage mass spectrometry (HPLC-DAD-ESI-MS (n) ) as well as by gas chromatography-mass spectrometry (GC-MS). The analytical workflow combining both methods revealed numerous compounds assigned for the first time as pineapple constituents by their mass fragmentations. Pineapple crown tissue was characterised by depsides of p-coumaric and ferulic acid. In contrast, major phenolic compounds in pineapple pulp extracts were assigned to diverse S-p-coumaryl, S-coniferyl and S-sinapyl derivatives of glutathione, N-L-γ-glutamyl-L-cysteine and L-cysteine, which were also identified in the peel. The latter was additionally characterised by elevated concentrations of p-coumaric, ferulic and caffeic acid depsides and glycerides, respectively. Two peel-specific cyanidin hexosides were found. Elevated concentrations of isomeric N,N'-diferuloylspermidines may be a useful tool for the detection of fraudulent peel usage for pineapple juice production. Mass fragmentation pathways of characteristic pineapple constituents are proposed, and their putative biological functions are discussed.

Analytical characterization of flavan-3-ol-rich apple juices produced with the innovative spiral filter press technology.

The retention of flavan-3-ols and other phenolic compounds during apple juice production at pilot plant scale (200 kg, cv. Boskoop) was investigated. An oxygen-excluding spiral filter press and a conventional decanter were used with and without pectinase mash treatment. Phenolic compounds were comprehensively identified and quantitated by RP-UHPLC and HILIC, both coupled to DAD-FLD and DAD-ESI(-)-QTOF-HR-MS/MS. These techniques combined with using a NIST cocoa flavan-3-ol standard allowed for the first time an individual quantification of flavan-3-ol fractions (DP 1-7) in apple juices. Spiral filter-pressed juices were exposed to less oxidation and exhibited four times higher total phenolic compound levels than decanter-made juices (1016 vs. 262 mg/L). Apple juices derived from pectinase-treated mashes had lower total phenolic compound levels than their non-treated counterparts. However, those made by spiral filter press still retained significantly higher concentrations (780 vs. 104 mg/L). Flavan-3-ols were especially well retained by spiral filter press processing, reaching unprecedentedly high concentrations of up to 713 mg/L. A 280 mL serving of non-treated spiral filter-pressed juice would therefore suffice to provide the daily intake of 200 mg flavan-3-ols, equaling the dose of cocoa flavan-3-ols associated with an authorized European health claim for healthy blood flow.

Characterisation of acylated anthocyanins from red cabbage, purple sweet potato, and Tradescantia pallida leaves as natural food colourants by HPLC-DAD-ESI(+)-QTOF-MS/MS and ESI(+)-MSn analysis.

Anthocyanins in red cabbage, sweet potato, and Tradescantia pallida leaves were characterised. A total of 18 non-, mono-, and diacylated cyanidins was identified in red cabbage by high performance liquid chromatography-diode array detection coupled to high-resolution and multi-stage mass spectrometry. Sweet potato leaves contained 16 different cyanidin- and peonidin glycosides being predominantly mono- and diacylated. In T. pallida leaves, the tetra-acylated anthocyanin tradescantin prevailed. The large proportion of acylated anthocyanins resulted in a superior thermal stability during heating of aqueous model solutions (pH 3.0) coloured with red cabbage and purple sweet potato extracts as compared to that of a commercial Hibiscus-based food dye. However, their stability was still outperformed by that of the most stable Tradescantia extract. Comparing vis spectra from pH 1-10, the latter had an additional, uncommon absorption maximum at approx. 585 nm at slightly acidic to neutral pH values, yielding intensely red to purple colours.

Effects of a plant-derived biostimulant application on quality and functional traits of greenhouse cherry tomato cultivars.

A greenhouse experiment was conducted to study the effects of the application of a plant-derived biostimulant (Bioup® TF) on fruit quality and composition of two clusters (cluster II and cluster VI) of the cherry tomato cultivars 'Eletta', 'Kaucana', and 'Top Stellina'. The biostimulant application promoted fruit yield by 12% (up to 1.3 kg m-2 in 'Kaucana') and increased the concentrations of important functional constituents like phytoene, γ-tocopherol and ß-tocopherol by up to 16, 25, and 23%, respectively. Fruits from late-ripe cluster VI showed higher fruit weights, D-fructose, and total sugar contents than those from early-ripe cluster II (by 15, 7 and 5%, respectively), but reduced concentrations of acyclic carotenoids (phytoene and lycopene) and tocochromanols (mainly γ-tocopherol, -44%). 'Top Stellina' showed the highest responsiveness to the biostimulant, as particularly (all-E)-ß-carotene, phytofluene, and γ-tocopherol concentrations increased, indicating a genotype-dependent effect of the treatment. However, fruits of all treated genotypes showed a contextual decrease in D-fructose and total sugars in response to the biostimulant (on average by 7 and 10%, respectively), indicating a metabolic load burdening the accumulation of lipophilic antioxidants in cherry tomatoes at the expense of their taste-related C pool.

Characterization of chromoplasts and carotenoids of red- and yellow-fleshed papaya (Carica papaya L.).

Chromoplast morphology and ultrastructure of red- and yellow-fleshed papaya (Carica papaya L.) were investigated by light and transmission electron microscopy. Carotenoid analyses by LC-MS revealed striking similarity of nutritionally relevant carotenoid profiles in both the red and yellow varieties. However, while yellow fruits contained only trace amounts of lycopene, the latter was found to be predominant in red papaya (51% of total carotenoids). Comparison of the pigment-loaded chromoplast ultrastructures disclosed tubular plastids to be abundant in yellow papaya, whereas larger crystalloid substructures characterized most frequent red papaya chromoplasts. Exclusively existent in red papaya, such crystalloid structures were associated with lycopene accumulation. Non-globular carotenoid deposition was derived from simple solubility calculations based on carotenoid and lipid contents of the differently colored fruit pulps. Since the physical state of carotenoid deposition may be decisive regarding their bioavailability, chromoplasts from lycopene-rich tomato fruit (Lycopersicon esculentum L.) were also assessed and compared to red papaya. Besides interesting analogies, various distinctions were ascertained resulting in the prediction of enhanced lycopene bioavailability from red papaya. In addition, the developmental pathway of red papaya chromoplasts was investigated during fruit ripening and carotenogenesis. In the early maturation stage of white-fleshed papaya, undifferentiated proplastids and globular plastids were predominant, corresponding to incipient carotenoid biosynthesis. Since intermediate plastids, e.g., amyloplasts or chloroplasts, were absent, chromoplasts are likely to emerge directly from proplastids.

Carotenoid profiles of red- and yellow-colored arils of cultivars of Taxus baccata L. and Taxus × media Rehder.

The botanical delimitation of Taxus species and cultivars may be facilitated by characterizing the pigment profiles of their red- and yellow-fleshed arils. Therefore, we determined genuine carotenoid profiles of differently colored arils of seven defined cultivars of Taxus baccata L. and Taxus × media Rehder. In-depth HPLC-DAD-ESI/APCI-MSn analyses revealed the presence of 43 carotenoids. Exceptional retro-carotenoids dominated the profiles of all samples assessed. Rhodoxanthin (E/Z)-isomers were predominant in the red-colored arils, resulting in a rather unusual abundance of carotenoid isomers as expressed by ratios of up to 1.3:2.0:0.9 between (all-E)-, (6Z)-, and (6Z,6'Z)-rhodoxanthin, respectively. By contrast, the uncommon yellow arils of Taxus baccata L. 'Lutea' mainly contained eschscholtzxanthin (E/Z)-isomers and esters. Total carotenoid concentrations ranged from 17.00 to 58.78 µg/g fresh weight across all samples assessed. Highest total rhodoxanthin concentrations of 51.33 ± 0.46 µg/g fresh weight were obtained from the red arils of Taxus × media Rehder 'Hicksii'. Overall, Taxus arils represent a promising source of carotenoids and, in particular, of retro-carotenoids with exceptional molecular structures and extraordinary absorption properties.

Coumarins, psoralens, and quantitative 1H-NMR spectroscopy for authentication of lemon (Citrus limon [L.] Burm.f.) and Persian lime (Citrus × latifolia [Yu.Tanaka] Tanaka) juices.

Mutual adulterations of lemon and lime juices may be detected using coumarins and psoralens as markers. Poor manufacturing practices or legal but mechanically intense processing of lemons were recently suspected to lead to false accusations of deliberate adulterations with lime juices due to potentially unspecific markers. Therefore, we studied coumarin and psoralen profiles in carefully dissected flavedo, albedo, and endocarp of lime and lemon as well as in juices produced under variable mechanical stresses at laboratory and pilot plant scale. Although the marker herniarin was detectable in juices from lime and harshly extracted lemons at low levels, isopimpinellin, bergapten and the herein proposed, tentatively assigned 5-geranyloxy-8-methoxypsoralen represented unambiguously lime-specific markers. Coumarin and psoralen data also allowed differentiating juices produced at differing degrees of mechanical stress. The latter was also possible using quantitative 1H-NMR spectroscopy, which yielded best results when combined with HPLC data on coumarins and psoralens. In the future, the reported approach may be used for establishing a robust database prior to being used in industrial practice.

Characterization of the Pale Yellow Petal/Xanthophyll Esterase gene family in citrus as candidates for carotenoid esterification in fruits.

In orange-pigmented citrus fruits, the xanthophyll esters are the predominant carotenoids, but their biosynthetic origin is currently unknown. In this work, seven PYP/XES (Pale Yellow Petal/ Xanthophyll esterase) genes were identified in Citrus genomes, but only PYP1-4 and 6 contained the structural domains essential for activity. The PYP/XES expression profiles in sweet orange and in other Citrus species such as lemon, mandarin and pummelo with marked differences in fruit pigmentation and content of xanthophylls esters, showed the upregulation of PYP1,2 and 6 genes during ripening only in orange-pigmented fruits. Moreover, transcript levels of PYP1, 2 and 6 genes in peel and pulp of sweet orange were accompanied by the accumulation of xanthophyll esters during ripening. This work reports for the first time the PYP/XES gene family in Citrus and strongly suggests its involvement in xanthophyll esterification in citrus fruit tissues and its influence in carotenoid accumulation and fruit pigmentation.

Evaluating the role of orange juice, HESPERidin in vascular HEALTH benefits (HESPER-HEALTH study): protocol for a randomised controlled trial.

INTRODUCTION: Although epidemiological studies associate the consumption of sugary beverages with adverse health effects, human experimental studies have demonstrated substantially different metabolic responses when 100% fruit juices are compared with artificial beverages. Fruit juices do not just provide sugars and associated calories, but they are also rich in bioactive compounds. Flavanones are bioactives specifically and abundantly found in citrus foods, with hesperidin as the major representative in sweet oranges. Flavanone intake has been associated with a lower incidence of mortality from cardiovascular disease (CVD). However, clinical evidence are too scarce to confirm the vasculoprotective effects of 100% orange juice (OJ) presumably mediated by flavanones and thereby do not allow firm conclusions to be drawn about their efficacy. METHODS AND ANALYSIS: The HESPER-HEALTH study aims to assess the efficacy of OJ in improving vascular function and the contribution of hesperidin to these effects. This double-blind, randomised, controlled, crossover study will be carried out in 42 volunteers predisposed to CVD, based on age and on overweight. It includes three 6-week periods of consumption of 330 mL/d of OJ versus control drinks with and without hesperidin at a dose in agreement with a daily OJ serving (approx. 200-215 mg). The primary outcome is endothelial function, assessed by flow mediated dilation, with measurements performed at fasting and postprandially in response to a challenge meal. The secondary outcomes include bioavailability and metabolism of flavanones, changes in other markers of vascular function, systemic biomarkers of cardiovascular risk, endothelial dysfunction and inflammation, vitamin C and carotenoids status, anthropometry and body composition, gut microbiota composition, nutrigenomic response and in oxylipin profiling. ETHICS AND DISSEMINATION: This ongoing study was approved by the Ethics committee Sud-Est III, Bron, France on 17 November 2020. The trial is registered on ClinicalTrials.gov. The results will be disseminated in peer-reviewed journals. TRIAL REGISTRATION NUMBER: NCT04731987; Pre-results.

Investigations into the Natural Occurrence of 1-Phenylethyl Acetate (Styrallyl Acetate).

So far, the occurrence of the flavor constituent 1-phenylethyl acetate in a natural source has not been unambiguously confirmed. The present work provides the detailed identification of 1-phenylethyl acetate from clove (Syzygium aromaticum (L.) Merr. & L.M. Perry) buds. In addition, headspace solid-phase microextraction-gas chromatography/mass spectrometry (GC/MS) analysis revealed further occurrence of 1-phenylethyl acetate in cocoa pulp and grape hyacinth flowers. A total of 15.2 g of essential oil was recovered from 7.2 kg of clove buds by simultaneous distillation-extraction followed by vacuum distillation. The distillate obtained was fractionated by silica column chromatography, whereby a significant enrichment of 1-phenylethyl acetate was achieved. The fraction containing the target analyte was further purified by preparative high-performance liquid chromatography, resulting in a final purity of ∼93.0%, yielding a total of 1 to 2 mg of 1-phenylethyl acetate. Identification of the isolated compound was achieved by GC/MS, infrared spectroscopy, enantioselective GC, isotope ratio MS, and nuclear magnetic resonance spectroscopy. Enantioselective GC/MS analysis revealed an enantiomeric excess of 60% (1S)-(-)-1-phenylethyl acetate in the isolate. The δ13CV-PDB value of -32.5 ± 0.5‰ was in accordance with that of C3-plants and other constituents found in genuine clove extracts.

(Poly)phenols, Carotenoids, and Tocochromanols in Corn (Zea mays L.) Kernels As Affected by Phosphate Fertilization and Sowing Time.

Corn (Zea mays L.) growth and development is often limited by the availability of phosphate. We thus hypothesized that phosphate fertilization may increase the contents of (poly)phenols, carotenoids, and tocochromanols (vitamin E) in corn grains. Corn plants cultivated on a soil fertilized with 44 kg phosphorus/ha were compared to plants grown on soil with low plant-available phosphate (1.6 mg CAL-P/100 g of soil), each sown early (April) and late (May) in a randomized field experiment. HPLC-DAD-(HR)-ESI-MSn revealed 19 soluble and 10 insoluble (poly)phenols, comprising phenolic acids, phenolic amines, diferulic, and triferulic acids in corn grains. Contents of individual (poly)phenols, carotenoids, and tocochromanols in whole grains were significantly (p < 0.05) increased by sowing time, but not by phosphate fertilization. In conclusion, low phosphate availability did not impair the biosynthesis of (poly)phenols, carotenoids, and tocochromanols in corn grains.

HPLC-DAD-APCI-MSn analysis of the genuine carotenoid pattern of pineapple (Ananas comosus [L.] Merr.) infructescence.

The genuine carotenoid pattern of pineapple infructescence was assessed by HPLC-DAD-APCI-MSn analysis. Prevailing pigments in the shell of 'MD2' (syn. "Extra Sweet") fruit were (all-E)-lutein and (all-E)-ß-carotene, in addition to chlorophylls a and b. The edible flesh contained (all-E)-violaxanthin, (all-E)-ß-carotene, and diverse esters of (9Z)-violaxanthin with caprylic, capric, lauric, and myristic acid. The latter esters have been reported for the first time as pineapple constituents. Total carotenoid concentrations in the edible fractions of the four varieties 'Sugar Loaf', 'Smooth Cayenne', 'MD2', and 'Queen Victoria' cultivated in Ghana ranged between 29 and 565 µg/100 g of fresh weight (FW). Total carotenoids in the flesh of fully ripe 'MD2' fruit exported by air freight amounted to 302 µg/100 g of FW, those in green ripe samples dispatched by sea freight to 359-432 µg/100 g of FW. All yellow fleshed cultivars exhibited a highly similar qualitative carotenoid profile.

Genuine Carotenoid Profiles in Sweet Orange [Citrus sinensis (L.) Osbeck cv. Navel] Peel and Pulp at Different Maturity Stages.

The carotenogenesis in the endocarp and flavedo of Navel oranges over four consecutive maturity stages was assessed by high-performance liquid chromatography-diode array detection-atmospheric pressure chemical ionization-multistage mass spectrometry. After optimization of the extraction method, 77 carotenoids, including 26 monoesters and 33 diesters of violaxanthin, ß-citraurin, and antheraxanthin, were characterized. Whereas chloroplast-specific pigments, such as (all-E)-lutein and (all-E)-ß-carotene, predominated in the flavedo of green-ripe fruit, a highly complex pattern of xanthophyll esters was found in the mature oranges. Total carotenoid contents of flavedo were approximately 9-fold higher [12 605 µg/100 g of fresh weight (FW)] than those in the endocarp (1354 µg/100 g of FW) at the fully mature stage. The mature endocarp abundantly contained violaxanthin mono- and diesters, in addition to diverse antheraxanthin esters, which were exclusively detected in this fruit fraction. Likewise, ß-citraurin esters were found to be unique flavedo constituents of mature fruit. Therefore, they may support the detection of fraudulent use of peel fractions during orange juice production.

Fatty acids, triacylglycerols, and thermal behaviour of various mango (Mangifera indica L.) kernel fats.

Mango processing generates high amounts of unexploited kernels. Lipid profiles and thermal behaviour of kernel fats from seven Mangifera indica L. cultivars originating from Latin America and Thailand were characterised. Total lipid contents ranged between 5.4 and 11.9%. Detailed GC-FID, GC-MS, and HPLC-DAD-ESI-MSn analyses revealed two fatty acids and five triacylglycerols as novel mango kernel constituents. Owing to the prevailing saturated fatty acids and triacylglycerols, 'Maha Chanook' fat had a significantly higher melting point, the highest solid fat indices at all temperatures, and densely packed crystals as shown by DSC and light microscopy, respectively. In contrast, 'Falan' exhibited high proportions of polyunsaturated fatty acids and triacylglycerols with low equivalent carbon numbers (40-46), resulting in lower melting and crystallisation temperatures and a loosened crystal network. 'Keitt' and 'Palmer' fats showed high proportions of triacylglycerols with medium equivalent carbon numbers (44-52). Mango kernels represent a sustainable source for liquid to semi-solid edible fats suitable for numerous potential applications, e.g., in food and cosmetics.

GC-MS profiling, descriptive sensory analysis, and consumer acceptance of Costa Rican papaya (Carica papaya L.) fruit purees.

Volatiles of papaya purees from four Costa Rican cultivars were analysed by solid-phase microextraction and gas chromatography-mass spectrometry. A total of 83 volatiles was assigned in the purees, of which 19 were detected for the first time as papaya constituents. As revealed by multivariate statistics, i.e., principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), the purees may be allocated to terpene- and lactone-rich ('SH-5'), ester-containing ('Criolla'), and intermediate chemotypes ('Pococí' and 'Silvestre'). Descriptive sensory analysis and a consumer acceptance test were additionally performed. Floral, fruity, and honey-like notes were perceived at significantly higher intensities in 'SH-5' puree. The latter descriptors strongly correlated with volatiles discriminating 'SH-5' in the PCA and PLS-DA, respectively. Consumer acceptance of the papaya purees differed significantly. 'Pococí' and 'SH-5' purees appear to be suitable for improving the nutritional value of blended fruit juices without impairing their sensory quality.

HPLC-DAD-APCI/ESI-MSn analysis of carotenoids and α-tocopherol in Costa Rican Acrocomia aculeata fruits of varying maturity stages.

Carotenoids and tocopherols were characterised in the meso- and exocarp of wild-growing Costa Rican Acrocomia aculeata fruits. Comprehensive profiling of these lipophilic micronutrients in fruits of three varying maturity stages was conducted for the first time. A method for the simultaneous extraction and quantitation of carotenoids and α-tocopherol was developed and validated. Detailed HPLC-DAD-APCI/ESI-MSn analyses enabled the identification of α-tocopherol and 25 carotenoids. The latter comprised antheraxanthin, ß-carotene, lutein, luteoxanthin, neoxanthin, phytoene, phytofluene, violaxanthin, zeaxanthin, and several (Z)-isomers of the aforementioned compounds. Quantitation by HPLC-DAD/FLD revealed total carotenoid concentrations of 872±178 and 3075±407µg/100g fresh weight in the meso- and exocarp of fully ripe fruits, respectively. In both fruit fractions, progressing maturation resulted in the accumulation of phytoene, phytofluene, (all-E)-zeaxanthin, (all-E)-antheraxanthin, and (all-E)-violaxanthin. Carotenoid profiling was supported by multivariate data analysis. Carotenoid precursors and xanthophyll cycle pigments characterised Macauba fruits of full maturity.

Characterization of Mesocarp and Kernel Lipids from Elaeis guineensis Jacq., Elaeis oleifera [Kunth] Cortés, and Their Interspecific Hybrids.

Morphological traits, total lipid contents, and fatty acid profiles were assessed in fruits of several accessions of Elaeis oleifera [Kunth] Cortés, Elaeis guineensis Jacq., and their interspecific hybrids. The latter featured the highest mesocarp-to-fruit ratios (77.9-78.2%). The total lipid contents of both E. guineensis mesocarp and kernel were significantly higher than for E. oleifera accessions. Main fatty acids comprised C16:0, C18:1n9, and C18:2n6 in mesocarp and C12:0, C14:0, and C18:1n9 in kernels. E. oleifera samples were characterized by higher proportions of unsaturated long-chain fatty acids. Saturated medium-chain fatty acids supported the clustering of E. guineensis kernels in multivariate statistics. Hybrid mesocarp lipids had an intermediate fatty acid composition, whereas their kernel lipids resembled those of E. oleifera genotypes. Principal component analysis based on lipid contents and proportions of individual fatty acids permitted clear-cut distinction of E. oleifera, E. guineensis, and their hybrids.

Authentication of pineapple (Ananas comosus [L.] Merr.) fruit maturity stages by quantitative analysis of γ- and δ-lactones using headspace solid-phase microextraction and chirospecific gas chromatography-selected ion monitoring mass spectrometry (HS-SPME-GC-SIM-MS).

Headspace solid phase microextraction and chirospecific gas chromatography-mass spectrometry in selected ion monitoring mode (HS-SPME-GC-SIM-MS) allowed quantitative determination of δ-lactones (δ-C8, δ-C10) and γ-lactones (γ-C6, γ-C8, γ-C10). A stable isotope dilution assay (SIDA) with d7-γ-decalactone as internal standard was used for quantitative analysis of pineapple lactones that was performed at three progressing post-harvest stages of fully ripe air-freighted and green-ripe sea-freighted fruits, covering the relevant shelf-life of the fruits. Fresh pineapples harvested at full maturity were characterised by γ-C6 of high enantiomeric purity remaining stable during the whole post-harvest period. In contrast, the enantiomeric purity of γ-C6 significantly decreased during post-harvest storage of sea-freighted pineapples. The biogenetical background and the potential of chirospecific analysis of lactones for authentication and quality evaluation of fresh pineapple fruits are discussed.