Quantitative Analysis of Glucosinolates in Oilseed Rape Based on HPLC of Desulfoglucosinolates and HPLC of Intact Glucosinolates

Bjerg, Birthe; Sørensen, Hilmer

doi:10.1007/978-94-009-3615-7_10

Cited by 51 publications

(30 citation statements)

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“…GSLs were desulfurized with sulfatase (Sigma) according to the method of Bjerg and Sørensen (1987), and desulfo-GSLs were subjected to HPLC, as reported in our previous study (Ishida et al, 2012). HPLC was performed on an LC-20A chromatograph (Shimadzu) fitted with a 5C 18-MS-II column (150 mm 3 4.6 mm i.d., 5 mm; Nacalai Tesque).…”

Section: Extraction and Hplc Analysis Of Gslsmentioning

confidence: 99%

A 2-Oxoglutarate-Dependent Dioxygenase Mediates the Biosynthesis of Glucoraphasatin in Radish

et al. 2017

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Glucosinolates (GSLs) are secondary metabolites whose degradation products confer intrinsic flavors and aromas to Brassicaceae vegetables. Several structures of GSLs are known in the Brassicaceae, and the biosynthetic pathway and regulatory networks have been elucidated in Arabidopsis (Arabidopsis thaliana). GSLs are precursors of chemical defense substances against herbivorous pests. Specific GSLs can act as feeding blockers or stimulants, depending on the pest species. Natural selection has led to diversity in the GSL composition even within individual species. However, in radish (Raphanus sativus), glucoraphasatin (4-methylthio-3-butenyl glucosinolate) accounts for more than 90% of the total GSLs, and little compositional variation is observed. Because glucoraphasatin is not contained in other members of the Brassicaceae, like Arabidopsis and cabbage (Brassica oleracea), the biosynthetic pathways for glucoraphasatin remain unclear. In this report, we identified and characterized a gene encoding GLUCORAPHASATIN SYNTHASE 1 (GRS1) by genetic mapping using a mutant that genetically lacks glucoraphasatin. Transgenic Arabidopsis, which overexpressed GRS1 cDNA, accumulated glucoraphasatin in the leaves. GRS1 encodes a 2-oxoglutarate-dependent dioxygenase, and it is abundantly expressed in the leaf. To further investigate the biosynthesis and transportation of GSLs in radish, we grafted a grs1 plant onto a wild-type plant. The grafting experiment revealed a leaf-to-root long-distance glucoraphasatin transport system in radish and showed that the composition of GSLs differed among the organs. Based on these observations, we propose a characteristic biosynthesis pathway for glucoraphasatin in radish. Our results should be useful in metabolite engineering for breeding of high-value vegetables.

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Section: Extraction and Hplc Analysis Of Gslsmentioning

confidence: 99%

A 2-Oxoglutarate-Dependent Dioxygenase Mediates the Biosynthesis of Glucoraphasatin in Radish

et al. 2017

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“…The intact glucosinolates and myrosinase isoenzymes (B. carinata cv. BRK-147-A) were from the laboratory collection [5,34,[46][47][48].…”

Section: Chemicals and Suppliesmentioning

confidence: 99%

“…These allelochemicals are present in all plants of the order Capparales [4] and have as well been reported in some few other species [1,5]. Myrosinase isoenzymes (EC 3.2.1.147) co-exist with glucosinolates and catalyze the hydrolysis of the ␤-d-thioglucopyranoside bond releasing an aglucone that further rearranges to a variety of products depending on the parent glucosinolate and the environmental conditions [6][7][8][9][10].…”

Section: Introductionmentioning

confidence: 97%

“…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%

“…The pHSA, the DSA and the SCEA are used in routine analysis of myrosinase activity of Brassica extracts due to their being simple, fast and inexpensive. However, despite their accuracy in the quantitative determination of the amount of glucosinolate degraded in assays based on only one glucosinolate, they are neither able to determine differences in the speed of degradation of glucosinolate mixtures nor the lack of degradation of thioglucoside substituted glucosinolates [3,5]. Furthermore, these methods do not allow determining the types of products formed and in which proportions they are produced.…”

Section: Introductionmentioning

confidence: 99%

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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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Identification and quantification of inorganic and organic selenium compounds with highperformance liquid chromatography

Kölbl

Kalcher

Irgolic

et al. 1993

Applied Organom Chemis

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Selenium appears in the natural selenium cycle in the form of several organic and inorganic compounds. The biologically beneficial and detrimental effects of 'selenium' must be ascribed to particular selenium compounds. The identification and quantification of selenium compounds in biological and environmental samples is required for an understanding of the role of selenium. The highperformance liquid-chromatographic (HPLC) methods for the separation, identification and quantification of selenite, selenate, hydrogen selenide, methaneselenol, bis(organothi0) selenides, trimethylselenonium salts, selenoamino-acids, selenium derivatives of carbohydrates, selenoproteins, selenonucleosides and other miscellaneous selenium compounds are summarized (193 references) and pertinent detection modes discussed. Advantages and disadvantages of the methods are pointed out. The literature is covered since 1974, the year of the first publication in this field.

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Quantitative Analysis of Glucosinolates in Oilseed Rape Based on HPLC of Desulfoglucosinolates and HPLC of Intact Glucosinolates

Cited by 51 publications

References 4 publications

A 2-Oxoglutarate-Dependent Dioxygenase Mediates the Biosynthesis of Glucoraphasatin in Radish

A 2-Oxoglutarate-Dependent Dioxygenase Mediates the Biosynthesis of Glucoraphasatin in Radish

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

Identification and quantification of inorganic and organic selenium compounds with highperformance liquid chromatography

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