Gas chromatography‐mass spectrometry of nitriles, isothiocyanates and oxazolidinethiones derived from cruciferous glucosinolates

Spencer, Gayland F.; Daxenbichler, M. E.

doi:10.1002/jsfa.2740310406

Cited by 150 publications

(78 citation statements)

References 5 publications

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“…Briefly, 100 mg of 4-week-old rosette tissue was homogenized in 1 mL of MES acid buffer (pH 6.0) and 0.4 mmol of allyl glucosinolate and incubated for 5 min. Glucosinolate activation products were extracted with dichloromethane for analysis by gas chromatography as described using a gas chromatographymass spectral detector (Agilent HP 6890 with an Agilent 5973N mass spectral detector), and peak identities for nitrile and isothiocyanate were established by comparison with published mass spectra (Spencere and Daxenbichler, 1980;Lambrix et al, 2001). Differences between the genotypes were tested using Student's t tests.…”

Section: Glucosinolate Structural Specificitymentioning

confidence: 99%

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

et al. 2009

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We identified an Arabidopsis (Arabidopsis thaliana) ethyl methanesulfonate mutant, modified vacuole phenotype1-1 (mvp1-1), in a fluorescent confocal microscopy screen for plants with mislocalization of a green fluorescent protein-d tonoplast intrinsic protein fusion. The mvp1-1 mutant displayed static perinuclear aggregates of the reporter protein. mvp1 mutants also exhibited a number of vacuole-related phenotypes, as demonstrated by defects in growth, utilization of stored carbon, gravitropic response, salt sensitivity, and specific susceptibility to the fungal necrotroph Alternaria brassicicola. Similarly, crosses with other endomembrane marker fusions identified mislocalization to aggregate structures, indicating a general defect in protein trafficking. Map-based cloning showed that the mvp1-1 mutation altered a gene encoding a putative myrosinase-associated protein, and glutathione S-transferase pull-down assays demonstrated that MVP1 interacted specifically with the Arabidopsis myrosinase protein, THIOGLUCOSIDE GLUCOHYDROLASE2 (TGG2), but not TGG1. Moreover, the mvp1-1 mutant showed increased nitrile production during glucosinolate hydrolysis, suggesting that MVP1 may play a role in modulation of myrosinase activity. We propose that MVP1 is a myrosinase-associated protein that functions, in part, to correctly localize the myrosinase TGG2 and prevent inappropriate glucosinolate hydrolysis that could generate cytotoxic molecules.

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Section: Glucosinolate Structural Specificitymentioning

confidence: 99%

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

et al. 2009

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“…Indirect methods involve the measurement of the released products, such as glucose [26] isothiocyanates and nitriles [1,2,14]. GC, LC and HPLC techniques for determination of glucosinolates and glucosinolate degradation products require in most cases more or less time consuming sample preparation steps prior to the chromatographic separation [3,5,14,[27][28][29]. Micellar electrokinetic capillary chromatography (MECC) has also been developed as a method of analysis for determination of glucosinolates, desulphoglucosinolates [30,31] and some of the glucosinolate degradation products, such as OZTs and nitriles [32][33][34].…”

Section: Introductionmentioning

confidence: 99%

Micellar electrokinetic capillary chromatography—Synchronous monitoring of substrate and products in the myrosinase catalysed hydrolysis of glucosinolates

Bellostas¹,

Sørensen²,

Sørensen³

2006

Journal of Chromatography A

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A micellar electrokinetic capillary chromatography (MECC) method has been developed for monitoring the myrosinase catalysed hydrolysis of 2-hydroxy substituted glucosinolates and the simultaneous formation of the corresponding degradation products (oxazolidine-2-thiones (OZTs) and nitriles). Glucosibarin ((2R)-2-hydroxy-2-phenylethylglucosinolate) was chosen as the model glucosinolate owing to the difficulties in determining hydrolysis rates of this type of substrates in traditional UV-assays. The method was afterwards validated with glucobarbarin ((2S)-2-hydroxy-2-phenylethylglucosinolate) and progoitrin ((2R)-2-hydroxybut-3-enylglucosinolate). Aromatic glucosinolates without a 2-hydroxy group in their side chains, such as glucotropaeolin (benzylglucosinolate) and gluconasturtiin (phenethylglucosinolate) were also tested. Formation of the glucosinolate hydrolysis products was monitored simultaneously at 206 nm and 230 nm. This allowed estimation of the extinction coefficient of the OZT derived from glucosibarin, which was found to be 18,000 M −1 cm −1 and 12,000 M −1 cm −1 at 206 nm and 230 nm, respectively. The developed method has limit of detection of 0.04 mM and 0.06 mM and limit of quantification of 0.2 mM and 0.3 mM for the glucosibarin derived OZT and nitrile, respectively. Linearity of the glucosinolate concentration was examined at six concentration levels from 2.5 mM to 100 mM and at 206 nm a straight line (R 2 = 0.9996) was obtained. The number of theoretical plates (N) at the optimal system conditions was 245,000 for the intact glucosibarin, 264,000 for the OZT and 252,000 for the nitrile.

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“…Injector and detector temperatures were 250 and 280˚C, respectively. Isopropyl, allyl, 4-pentenyl, and phenethyl isothiocyanates were identified by GC/MS with authentic samples and data of a reference (Spencer & Daxenbichler, 1980).…”

Section: Methodsmentioning

confidence: 99%

Changes in Pungent Components of Two Wasabia japonica MATSUM. Cultivars during the Cultivation Period

Hara

Mochizuki

Kaneko

et al. 2003

FSTR

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Japanese horseradish, i.e. wasabi (Wasabia japonica MATSUM.), cultivars are classified as green-stem and red-stem types. It is believed that pungency differs slightly between the types. We analyzed isothiocyanate levels and myrosinase activity in two wasabi cultivars, Maruichi and Mazuma, a green-stem and a red-stem type, respectively. Time-course experiments were performed monthly from May 2001 to October 2002, separating plants into leaves, petioles, and rhizomes. Allyl isothiocyanate production induced by maceration occurred in whole plants in both cultivars, though the major organ producing the isothiocyanate was the rhizome. Levels of myrosinase activity in the rhizome of Maruichi increased in November and December 2001, when allyl isothiocyanate production also increased in Maruichi's rhizome. Although levels of allyl isothiocyanate did not differ between the two cultivars, Mazuma produced isopropyl isothiocyanate but Maruichi produced little. These results suggest that the difference in pungency between Maruichi and Mazuma may be derived from the isothiocyanate component.

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Gas chromatography‐mass spectrometry of nitriles, isothiocyanates and oxazolidinethiones derived from cruciferous glucosinolates

Cited by 150 publications

References 5 publications

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

MODIFIED VACUOLE PHENOTYPE1 Is an Arabidopsis Myrosinase-Associated Protein Involved in Endomembrane Protein Trafficking

Micellar electrokinetic capillary chromatography—Synchronous monitoring of substrate and products in the myrosinase catalysed hydrolysis of glucosinolates

Changes in Pungent Components of Two Wasabia japonica MATSUM. Cultivars during the Cultivation Period

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