Betaines in fruits of Citrus genus plants

Numerous compounds, many of them osmolytes, were quantified in natural juices and in frozen concentrate juices from fruits of plants of the Citrus genus. L-proline, N-methyl-L-proline (hygric acid), N,N-dimethyl-L-proline (stachydrine), 4-hydroxy-L-prolinebetaine (betonicine), 4-hydroxy-L-proline, γ-aminobutyric acid (Gaba), 3-carboxypropyltrimethylammonium (GabaBet), N-methylnicotinic acid (trigonelline), and choline in the fruit juices of yellow orange, blood orange, lemon, mandarin, bitter orange (Citrus aurantium), chinotto (Citrus myrtifolia), and grapefruit were analyzed by sensitive HPLC-ESI-tandem mass spectrometry procedure. It was found that the most represented osmolytes in the juices, that is, L-proline, stachydrine, and betonicine, can be quantified with minimal sample preparation and short analysis time (about 1 min) also by flow injection analysis (FIA) ESI-MS/MS with the same results as obtained by HPLC ESI-MS/MS. In all of the juices, discrete amounts of choline and trigonelline were present. Conversely, GabaBet was always below detection limits. Notably, N-methyl-L-proline and 4-hydroxy-L-prolinebetaine, which were discovered for the first time in the juice of bergamot (Citrus bergamia Risso et Poit), are also present in all of the citrus juices examined.     

Essential oil composition of Citrus meyerii Y. Tan. and Citrus medica L. cv. Diamante and their lemon hybrids

In this paper we report the volatile fraction composition of Citrus meyerii Y. Tan. and Citrus medica L. cv. Diamante and two new lemon hybrids obtained by cross-breeding them with the tetraploid Citrus limon Burm. cv. Cavone. Both parent and hybrid oils were laboratory-extracted from the peel fruits and analyzed by HRGC-MS and HRGC-FID. Sixty-three components were fully characterized by mass spectra, linear retention indices, and injection of standards. The average composition as single components for all the oils analyzed is reported. Moreover, the data obtained were statistically analyzed. Since limonene is by far the main component of all the essential oils examined, analysis of variance and multivariate analysis gave interesting information on the similarities and differences between the oils analyzed. The new hybrid oils analyzed have potential commercial value because they could be an acceptable alternative to the valuable lemon oil.     

Characterization of anthocyanins and pyranoanthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juice

Anthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juices were isolated and purified by means of high-speed countercurrent chromatography and preparative high-performance liquid chromatography. Structures of the pigments were then elucidated by electrospray ionization multiple mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. The major anthocyanins of the juice were characterized as cyanidin 3-glucoside and cyanidin 3-(6"-malonylglucoside). Furthermore, six minor anthocyanins were detected and identified as cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, delphinidin 3-(6"-malonylglucoside), peonidin 3-(6"-malonylglucoside), and cyanidin 3-(6"-dioxalylglucoside). The occurrence of the latter compound in blood oranges is reported here for the first time, together with full NMR spectroscopic data. Further investigations revealed the presence of four anthocyanin-derived pigments, which are formed through a direct reaction between anthocyanins and hydroxycinnamic acids during prolonged storage of the juice. These novel pyranoanthocyanins were identified as the 4-vinylphenol, 4-vinylcatechol, 4-vinylguaiacol, and 4-vinylsyringol adducts of cyanidin 3-glucoside through comparison of their mass spectrometric and chromatographic properties with those of synthesized reference compounds.     

Composition and seasonal variation of the essential oils from two mandarin cultivars of southern Brazil

Cold-pressing and hydrodistilled peel oils of two Brazilian mandarin cultivars, commonly called Cai and Montenegrina (Citrus deliciosa Tenore), were obtained from fruits collected on mandarin trees submitted to the same pedoclimatic and cultural conditions. Their chemical composition and seasonal variation of the main volatile constituents were investigated by capillary GC and GC-MS, and the results were submitted to statistical analysis. To better characterize the oils, the enantiomeric distribution ratio of seven components (alpha-pinene, sabinene, beta-pinene, limonene, linalool, terpinen-4-ol, and alpha-terpineol) was determined by multidimensional gas chromatography (MDGC). The similarities found between both C. deliciosa cultivars are strong evidence for the hypothesis that Montenegrina originated from Cai as previously reported from agronomic studies.     

基于低场核磁共振的柑橘汁胞粒化评级

为更加准确、客观、快速地评定柑橘汁胞粒化程度,改变目前主要依靠肉眼感官评级的现状。该文进行了机器打分研究,并与感官评价进行了相关分析。该研究将低场核磁成像及磁弛豫时间测试结果与柑橘汁胞含水率及汁胞粒化程度的感官分级等指标相对应,应用于柑橘汁胞粒化程度的人工智能鉴定和识别。相关分析、F检验和多重比较等数学分析表明,利用低场核磁横向弛豫时间的响应值可有效反应汁胞粒化程度。横向弛豫时间3个峰的响应值与不同汁胞粒化程度柑橘的感官评价等级及汁胞含水率、果实失重率等存在显著相关(P0.05)或极显著相关(P0.01)。经成对t检验分析,基于低场核磁建立的汁胞粒化评级模型与评价员得到感官平均评分的评价结果间没有显著差异(P=0.4982t=0.7143)。研究结果为柑橘汁胞粒化程度感官分级的理化技术替代研究提供了有益尝试。     

生长期喷钙提高锦橙果实品质及延长贮藏期

探讨钙对柑橘果实贮藏品质及衰老的影响,为合理调控柑橘钙素营养、延长果实的贮藏保鲜期提供理论和技术支撑。通过在北碚447锦橙(Citrus sinensis Osbeck cv.Jincheng)生长的不同时期进行树体补钙,研究钙对果实贮藏品质及酶活性的影响。结果表明,在锦橙的生长期喷钙能提高果实钙含量,抑制果实贮藏过程中维生素C等物质的氧化分解,提高果实可溶性固形物含量和糖酸比,改善果实品质。不同时期喷钙均能提高果实中抗氧化酶(CAT、SOD)活性,降低细胞壁水解酶(PG、CX)、过氧化物酶(POD)活性,减轻脂质过氧化程度,果胶的分解转化速度减慢,丙二醛、可溶性果胶的含量明显降低,从而维持果皮具有一定的强度,延缓了果实的衰老,降低烂果的发生率,延长了果实的贮藏保鲜期。其中以幼果期喷钙在提高果实钙含量、延长贮藏期等方面的效果较好,其次是果实膨大期喷钙,成熟期喷钙效果最差。在幼果期和果实膨大期喷钙是提高果实品质、延长采后果实贮藏保鲜期的重要措施。     

Identification and quantification of carotenoids, by HPLC-PDA-MS/MS, from Amazonian fruits

The major and minor carotenoids from six fruits, buriti (Mauritia vinifera), mamey (Mammea americana), marimari (Geoffrola striata), peach palm (Bactrys gasipaes), physalis (Physalis angulata), and tucuma (Astrocaryum aculeatum), all native to the Amazonia region, were determined by high-performance liquid chromatography-photodiode array detector-mass spectrometry detector (HPLC-PDA-MS/MS), fulfilling the recommended criteria for identification. A total of 60 different carotenoids were separated on a C30 column, all-trans-beta-carotene being the major carotenoid found in all fruits. The presence of apo-10'-beta-carotenol, found in mamey, was not previously reported in foods. In addition, this is the first time that the identification of beta-zeacarotene in natural sources is supported by MS data. The total carotenoid content ranged from 38 microg/g in marimari to 514 microg/g in buriti. All fruits analyzed can be considered good sources of provitamin A, especially buriti, with 7280 RE/100 g.     

Occurrence of pipecolic acid and pipecolic acid betaine (homostachydrine) in Citrus genus plants

The presence of pipecolic acid and pipecolic acid betaine, also known as homostachydrine, is herein reported for the first time in Citrus genus plants. Homostachydrine was found in fruits, seeds, and leaves of orange, lemon, and bergamot (Citrus bergamia Risso et Poit). As homostachydrine was not commercially available, as a comparative source, extracts of alfalfa leaves ( Medicago sativa L.) were used, in which homostachydrine is present at high concentration. Then, the results where confirmed by comparison with an authentic standard synthesized and purified starting from pipecolic acid. The synthesized standard was characterized by a ESI-MS/MS study using a 3D ion-trap mass spectrometer. When subjected to MS/MS fragmentation in positive ion mode, homostachydrine, unlike its lower homologue proline betaine (also known as stachydrine), showed a pattern of numerous ionic fragments that allowed unambiguous identification of the compound. For the quantitation in the plant sources, high sensitivity and specificity were achieved by monitoring the transition (158 → 72), which is absent in the fragmentation patterns of other major osmolytes commonly used as markers for studies of abiotic stress. As for the metabolic origin of homostachydrine, the occurrence in citrus plants of pipecolic acid leads to the hypothesis that it could act as a homostachydrine precursor through direct methylation.     

柑桔皱皮果与果皮及其细胞壁矿质元素关系的研究

对红 1 7甜橙、红江橙及红江橙正常果与皱皮果、红江橙阴面果与阳面果的果皮及其细胞壁N、P、K、Ca、Mg的含量进行了分析 ,探讨这些矿质元素与柑桔成熟期皱皮果形成的关系。结果发现红 1-7甜橙皱皮果发生率较低 ,仅为 7.34% ,而红江橙皱皮果发生率高达 28.06% ;皱皮现象 86%以上出现在果实的阴面果皮。果皮N、P含量与柑桔皱皮果的形成没有相关性 ,K在皱皮果中的含量比正常果高 ,而在阳面果皮比阴面果皮高 ;Mg在皱皮果和阴面果皮及其细胞壁的含量都比在正常果和阳面果皮稍低 ,但Mg的含量与皱皮果率的相关性不显著 ;Ca在正常果和阳面果皮及其细胞壁的含量都明显高于皱皮果和阴面果皮 ,皱皮果率与Ca的含量呈显著的负相关     

Composition of the essential oil of Citrus tamurana Hort. ex Tanaka (Hyuganatsu)

The composition of the essential oil of Citrus tamurana Hort. ex Tanaka (Hyuganatsu), isolated by the cold-pressing method, was investigated by capillary GC and GC-MS. The effects of harvesting time, degree of freshness, and size of fruits on the composition of Hyuganatsu peel oils were also determined. A total of 126 volatile constituents were confirmed in the Hyuganatsu oils. The Hyuganatsu oils contained hydrocarbons (95.95-96.95%), aldehydes (0.33-0.62%), alcohols (1.91%-2.64%), ketones (0.40-0.62%), esters (0.28-0.39%), oxides (0.04-0.06%), acids (0.01%), and trace amounts of fugenol methyl ether. Monoterpene hydrocarbons were predominant. Limonene (80.35-82.39%), gamma-terpinene (7.71-9.03%), myrcene (2.11-2.28%), linalol (1.37-2.01%), and alpha-pinene (1.17-1.43%) were the most abundant components in Hyuganatsu oils. The principal sesquiterpene hydrocarbon was trans-beta-farnesene (0.60-1.04%), and its content in Hyuganatsu oils was higher than in oils of other citrus fruits. The number of ketones and the content of l-carvone in Hyuganatsu oils were higher than in other citrus oils.     

Changes in the levels of polymethoxyflavones and flavanones as part of the defense mechanism of Citrus sinensis (cv. Valencia Late) fruits against Phytophthora citrophthora

Phytophthora citrophthora causes serious losses in Citrus fruits through brown rot lesion. The effect of infection with P. citrophthora on Citrus sinensis (cv. Valencia Late) fruits was studied, with particular reference to the levels of the flavanones hesperidin and isonaringin and the polymethoxyflavones sinensetin, nobiletin, tangeretin, and heptamethoxyflavone, because flavonoids are most probably involved as natural defense or resistance mechanisms in this genus. Changes in the levels of these flavonoids were detected after infection. The hesperidin and isonaringin contents fell by 13 and 67%, respectively, whereas the contents of their corresponding aglycons, hesperetin and naringenin, increased, suggesting the hydrolyzing effect of this fungus on the glycosylated flavanones. The heptamethoxyflavone, nobiletin, sinensetin, and tangeretin levels increased by 48, 28, 26, and 24%, respectively. The in vitro study revealed that these compounds acted as antifungal agents, the most active being the aglycons (naringenin and hesperetin), followed by the polymethoxyflavones and flavanone glycosides. The participation of these flavonoids in the defense mechanism of this Citrus species is discussed.     

Use of nootkatone as a senescence indicator for Rouge La Toma Cv. grapefruit (Citrus paradisi Macf.)

The objective of this research was to study the usefulness of nootkatone as a senescence indicator for Rouge La Toma cv. grapefruit (Citrus paradisi Macf.), simulating different treatments that included the normal postharvest handling of citrus fruits: temperature conditioning, cold storage, shipment periods to overseas markets such as Japan and the U.S., marketing conditions, and storage at nonchilling temperature (control treatments). The highest nootkatone levels, determined by GLC-MS analyses, were detected in fruits subjected to control treatments. No significant differences were observed in nootkatone levels between treatments either with or without temperature conditioning prior to the start of the cold storage. Levels of nootkatone increased throughout time for all assayed treatments. The linear regressions of nootkatone levels showed correlation coefficients of 0.80 and 0.83 with storage time (29 and 42 days, respectively). Therefore, nootkatone appears to be a good indicator of senescence for Rouge La Toma grapefruit.     

Stable isotope and trace element profiling combined with classification models to differentiate geographic growing origin for three fruits: effects of subregion and variety

Classifications of geographic growing origin of three fresh fruits combining elemental profiles with various modeling approaches were determined. Elemental analysis (Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, V, and Zn) of strawberry, blueberry, and pear samples was performed using inductively coupled plasma argon atomic emission spectrometer. Bulk stable carbon and nitrogen isotope analyses in pear were performed using mass spectrometry as an alternative fingerprinting technique. Each fruit, strawberry (Fragaria x ananassa), blueberry (Vaccinium caesariense/corymbosum), and pear (Pyrus communis), was analyzed from two growing regions: Oregon vs Mexico, Chile, and Argentina, respectively. Principal component analysis and canonical discriminant analysis were used for data visualization. The data were modeled using linear discriminant function, quadratic discriminant function, neural network, genetic neural network, and hierarchical tree models with successful classification ranging from 70 to 100% depending on commodity and model. Effects of Oregon subregional and variety classification were investigated with similar success rates.     

沙田柚开花座果期子房内源激素变化研究

对开花座果期不同授粉处理的沙田柚子房内源吲哚乙酸、玉米素、脱落酸、赤霉素进行了定量测定。结果表明,不同授粉处理．子房内源激素的变化出现明显差异,在酸柚异花授粉的子房中,生长促进物质的含量水平明显高于自花授粉的子房,而生长抑制物质的变化则相反,认为这种激素的动态变化差异,是沙田柚自花不实的主要原因。     

Characterization and quantitation of antioxidant constituents of sweet pepper (Capsicum annuum L.)

Sweet peppers (Capsicum annuum L.) cv. Vergasa have been studied at four maturity stages (immature green, green, immature red, and red). The individual phenolics (hydroxycinnamic acids and flavonoids), vitamin C (ascorbic acid and dehydroascorbic acid), and individual carotenoids were characterized and quantified. Five hydroxycinnamic derivatives and 23 flavonoids were characterized and quantified from the pericarp of sweet pepper by high-performance liquid chromatography-diode array detection-electrospray ionization-mass spectrometry. Identification was carried out by their UV spectra, chromatographic comparisons with authentic markers, identification of hydrolysis products, and tandem mass spectrometry analysis. Hydroxycinnamic derivatives, O-glycosides of quercetin, luteolin, and chrysoeriol, and a large number of C-glycosyl flavones have been characterized. Some of these compounds were found for the first time in nature. Clear differences in the individual and total phenolic content were detected between the different maturity stages. Immature green pepper had a very high phenolic content while green, immature red, and red ripe peppers showed a 4-5-fold reduction. Ascorbic acid was the main form of vitamin C, and its content increased as the pepper reached maturity. The red ripe stage had a relevant impact on the carotenoids content. Thus, immature green peppers showed the highest content of polyphenols, while red ripe fruits had the highest content of vitamin C and provitamin A.     

Characterization of the geographical origin of Pecorino Sardo cheese by casein stable isotope ((13)c/(12)c and (15)n/(14)n) ratios and free amino acid ratios

The stable isotope ratios ((13)C/(12)C and (15)N/(14)N) of casein measured by isotope ratio mass spectrometry (IRMS) and some free amino acid ratios (His/Pro, Ile/Pro, Met/Pro, and Thr/Pro) determined by HPLC in samples of ewes' milk cheese from Sardinia, Sicily, and Apulia were found to be parameters independent of ripening time. Multivariate data treatments performed by applying both unsupervised (principal component analysis and cluster analysis) and supervised [linear discriminant analysis (LDA)] methods revealed good discrimination possibilities for the cheeses according to place of origin. In this respect, particularly significant were the variables Ile/Pro, Thr/Pro, (13)C/(12)C, and (15) N/(14)N ratios on which basis 100% discrimination and classification of the samples by LDA was obtained.     

动物源食品中兽药残留高通量快速分析检测技术

随着养殖业的迅猛发展,动物源食品兽药残留问题日益成为食品安全领域的重要内容。动物源食品基质复杂,而其中残留的兽药含量甚微,传统的样品制备及检测方法大多存在检测样品基质种类单一、检测兽药种类范围小、耗时长、重现性差等问题,缺乏一定的通用性和准确性,已不能满足当前社会发展的需要。近年来,随着QuChERS法和高分辨质谱等先进样品制备和检测技术在兽药残留检测分析领域的应用,一批高通量、自动化乃至可视化的快速高效分析检测方法也随之而起,该文即对这些高通量快速样品制备和检测方法进行综述,同时对动物源食品中兽药残留的检测和监控等工作提出建议并进行了展望。     

Compound specific plant amino acid δ15N values differ with functional plant strategies in temperate grassland

The distribution and natural abundance isotopic (δ¹⁵N) content of whole tissue and individual amino acids in plants in a temperate grassland were determined using ion chromatography (IC), continuous flow‐isotope ratio mass spectrometry (CF‐IRMS), and gas chromatography‐combustion‐isotope ratio mass spectrometry (GC‐C‐IRMS). The results showed that the selected plants (Lolium perenne, Juncus effusus, and Brachythecium rutabulum) differed in their amino acid content and distribution from the parent grassland soil. Bulk and individual amino acid δ¹⁵N isotope signatures were different between the plants, which concurred with their functional strategy in relation to the relative acquisition of available N sources. The individual amino acid δ¹⁵N values of histidine and phenylalanine could be used to differentiate between the three plant species.     

Nutrient Indexing in “‘Kinnow” Mandarin (Citrus deliciosia Lour.× Citrus nobilis Tanaka) Grown in Indogangetic Plains

Diagnosis and remediation of nutrient constraints in perennial fruit crop like citrus are the two important pillars of an effective nutrient management program. Efforts were made to develop nutrient indexing (NI) criteria based on generated leaf and soil analysis dataset for “Kinnow” mandarin (Citrus deliciosia Lour. × Citrus nobilis Tanaka) grown on illitic soils of Indogangetic plains (Entisol, Inceptisol, and Aridisol). NI through diagnosis and recommendation integrated system (DRIS) using leaf analysis data showed optimum value of leaf nutrient concentration as 2.22–2.32% nitrogen (N), 0.11–0.15% phosphorus (P), 1.10–1.41% potassium (K), 2.32–2.79% calcium (Ca), 0.38–0.61% magnesium (Mg), 22.4–58.3 ppm iron (Fe), 26.3–56.2 ppm manganese (Mn), 4.2–7.2 ppm copper (Cu), and 21.3–26.9 ppm zinc (Zn) vis-à-vis a fruit yield of 32.4–56.1 kg tree^?1. Using these NI criteria, Zn was observed as most deficient (64.7%) followed by Fe (61.5%), Mn (57.6%), N (96.1%), and P (38.5%) using percentage of orchards as basis. While, optimum NI (mg kg^?1) using soil analysis data was determined as 114.3–121.2 potassium permanganate (KMnO₄-N), 7.8–12.3 Olsen-P, 96.4–131.3 ammonium acetate (NH₄OAc)-K, 189.4–248.6 NH₄OAc-Ca, 72.3–89.9 NH₄OAc-Mg, 5.8–11.1, DTPA-Fe, 4.3–6.9 diethylenetriaminepentaacetic acid (DTPA)-Mn, 0.45–0.69 DTPA-Cu, and 21.3–26.9 DTPA-Zn for the optimum yield of 32.4–56.1 kg tree^?1. Soil analysis-based NIs displayed a good complementary with leaf analysis-based NIs evident from the diagnoses indicating Mn (52.2%) as most dominant constraint Zn (61.2%) followed by Mn (48.3%), N (41.2%), and P (35.6%). The recommended DRIS-based NIs would lay a scientific basis in formulating citrus fertilization program.     

Chemometric characterization of fruit juices from Spanish cultivars according to their phenolic compound contents: I. Citrus fruits

The data set composed by phenolic compound profiles of 83 Citrus juices (determined by HPLC-DAD-MS/MS) was evaluated by chemometrics to differentiate them according to Citrus species (sweet orange, tangerine, lemon, and grapefruit). Cluster analysis (CA) and principal component analysis (PCA) showed natural sample grouping among Citrus species and even the Citrus subclass. Most of the information contained in the full data set can be captured if only 15 phenolic compounds (concentration ≥10 mg/L), which can be quantified with fast and accurate methods in real samples, are introduced in the models; a good classification which allows the confirmation of the authenticity of juices is achieved by linear discriminant analysis. Using this reduced data set, fast and routine methods have been developed for predicting the percentage of grapefruit in adulterated sweet orange juices using principal component regression (PCR) and partial least-squares regression (PLS). The PLS model has provided suitable estimation errors.