Isolation of Intact Glucosinolates by Column Chromatography and Determination of Their Purity

Bjerg, Birthe; Sørensen, Hilmer

doi:10.1007/978-94-009-3615-7_6

Cited by 17 publications

(12 citation statements)

References 7 publications

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“…The crude extract was evaporated to dryness and redisolved in 5 mL of Milli‐Q water. Desulphoglucosinolates were produced following the standard procedure6 and analytically determined by micellar electrokinetic capillary chromatography (MECC) 30. The MECC data have been found to give the same results as HPLC29,31 when both sets of data are corrected with the same response factors published (ISO 9167‐1) 32.…”

Section: Methodsmentioning

confidence: 98%

“…Freeze‐dried plant material (300–500 mg, composite of three replicate plants) was extracted following the method described elsewhere 6. Two extractions (2 min each) of the material were conducted in 3 mL of 70% boiling methanol, with a solution of glucotropaeolin and glucosibarin (100 µL of 7 µmol L −1 each) added as internal standards before starting the extractions 29.…”

Section: Methodsmentioning

confidence: 99%

“…Glucosinolates are alkyl‐ N ‐hydroximine sulphate esters with a β‐ D ‐thioglucopyranoside group attached to the hydroximine carbon in Z ‐configuration to the sulphate group 1–3. These natural products are biosynthetically produced from amino acids and are present in all plants of the order Capparales and in a few other plants 1,4–6. Most glucosinolate‐containing genera are clustered within the families Capparaceae, Caricaceae and Brassicaceae, the latter including the genus Brassica .…”

Section: Introductionmentioning

confidence: 99%

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Profiling glucosinolates in vegetative and reproductive tissues of four Brassica species of the U‐triangle for their biofumigation potential

Bellostas

Sørensen

2007

J Sci Food Agric

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Glucosinolates are amino acid derived allelochemicals present in all plants of the order Capparales. These compounds are degraded by myrosinase isoenzymes, releasing a series of biologically active products defined by the parent glucosinolate and the reaction conditions. Species within the Brassicaceae are found to differ in their glucosinolate profile and glucosinolate concentrations. Different tissues within a single plant also show such variations, which are further influenced by the growth stage and environmental conditions. In the experiments described in this paper, four Brassica species of the U-triangle (B. carinata, B. nigra, B. juncea and B. rapa) were compared with respect to their glucosinolate profiles in roots, stems, leaves and reproductive organs at different developmental stages. The glucosinolate profile of corresponding ripe seeds was also determined. Prop-2-enylglucosinolate was identified as the major glucosinolate in the three mustards, where it represented over 90% of the total glucosinolate concentration of ripe seeds and over 50% of green tissues. The relative concentration of this glucosinolate increased in all tissues during plant growth. Brassica rapa showed a different glucosinolate profile than the three mustards, with higher concentrations of but-3-enylglucosinolate, 2-hydroxybut-3-enylglucosinolate and 2-hydroxypent-4-enylglucosinolate. The concentration of indol-3-ylmethylglucosinolates was also higher in B. rapa than in the mustard plants, with 4-hydroxyglucobrassicin representing 30% of the total glucosinolate concentration in ripe seeds. The total glucosinolate concentration of the species studied varied with growth stage and the mustards achieved a maximum towards the end of the period monitored. Glucosinolate concentration decreased in roots and leaves but increased in reproductive tissues. The determined glucosinolate profiles are an initial step in assessing the biofumigation potential of these species of the Brassicaceae family.

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Section: Methodsmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Profiling glucosinolates in vegetative and reproductive tissues of four Brassica species of the U‐triangle for their biofumigation potential

Bellostas

Sørensen

2007

J Sci Food Agric

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“…Benzylglucosinolate and sinalbin were from the laboratory collection [Bjerg & Sørensen, 1987] as well as other pro analyses chemicals . …”

Section: Chemicalsmentioning

confidence: 99%

Effects of Novel Processing Techniques on Glucosinolates and Membrane Associated Myrosinases in Broccoli

Frandsen¹,

Markedal²,

Martı́n-Belloso³

et al. 2014

Pol. J. Food Nutr. Sci.

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longed shelf-life by inactivation of micro-organisms and enzymes. HP/HT can at temperatures above 50 o C lead to pasteurization using pressure levels from 250-500 MPa [Wilson et al., 2008], while sterilization can be obtained with HP at temperatures higher than 70 o C . Focus is thus on the application of HP in combination with high temperatures (HT), which may lead to tissue compression, cell wall breakage and cell membrane disruption, depending on the HP and HT levels as well as the type of vegetable or fruit that are treated [Michel & Autio, 2002]. In the present study, where broccoli has been processed, the food quality is particularly connected with the enzyme-substrate (myrosinase--glucosinolate) system, where the different types of non-enzymatic and enzymatic catalyzed hydrolysis products give rise to effects from being health promoting to affecting the taste and odor [Hansen et al., 1995[Hansen et al., , 1997 Jeffery & Aray, 2009].PEF is considered a non-thermal treatment, which can lead to pasteurization at ambient temperatures using electrical field strengths of 20-30 kV/cm, and short pulses of 1-10 µs [Mosqueda-Melgar et al., 2008]. Application of short electric pulses may result in pore formation in cell membranes, and depending on the treatment intensity, time, electric field and pulse energy the membrane permeabilisation High pressure/high temperature (HP/HT) and pulsed electric field (PEF) treatment of food are among the novel processing techniques considered as alternatives to conventional thermal food processing. Introduction of new processing techniques with fast and gentle processing steps may reveal new possibilities for preservation of healthy bioactive compounds in processed food. However, effects on various food components due to autolysis and fast reactions prior to the applied HP/HT or PEF need to be considered as the total contribution of processing steps affects the obtained food quality. The present experiments were performed on broccoli (Brassica oleracea var. Italica) florets, purée and juice. Specific focus was given to effects of HP/HT and PEF processing on the content of glucosinolates and activities of myrosinase isoenzymes (EC.3.2.1.147) in the broccoli preparations. Certain conditions applied in HP/HT processing of broccoli florets were able to maintain a high level of intact glucosinolates. Treatment at 700 MPa and 20°C for 10 min was found to inactivate myrosinase activity, but also pressure treatments at 300 MPa and 20°C were able to maintain a high level of intact glucosinolates present in the untreated broccoli florets. PEF processing of broccoli purée and juice showed that the myrosinase activities resulted in nearly total glucosinolate transformations as result of autolysis during puréeing and juice making prior to the PEF processing. These data demonstrated that insight into potential effects on myrosinase activities from application of PEF processing implies specific focus on the sample steps preceding the PEF processing. Processing of broccoli and autolysis reaction...

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“…As an internal standard, 0.3 μmol of sinigrin (Sigma‐Aldrich, Stockholm, Sweden) or glucotropeaolin (Merck KgaA, Darmstadt, Germany) was added. The samples were centrifuged for 5 min at 4 000 rpm (2 880 g ) (Megafuge 1.0, Heraeus Sepatech, Osterode, Germany) and the entire supernatant was subsequently added to a DEAE Sephadex A‐25 column (Pharmacia, Uppsala, Sweden) and desulphorylated by incubating with Helix pomatia sulphatase (Sigma‐Aldrich) over night, according to Bjerg and Sørensen (1987). After elution from the column using 3×300 μl sterile water, the samples were concentrated to dryness followed by resuspension in 40 μl of water.…”

Section: Methodsmentioning

confidence: 99%

B‐genome derived resistance to Leptosphaeria maculans in near isogenic Brassica napus lines is independent of glucosinolate profile

Wretblad

Dixelius

2000

Physiologia Plantarum

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The role of the glucosinolate‐myrosinase system in resistance to Leptosphaeria maculans was studied by monitoring changes in glucosinolate profiles in leaf tissue surrounding the site of inoculation. Susceptible Brassica napus cv. Hanna, resistant B. nigra and near isogenic lines derived from interspecific hybrids between the two species were compared. Expression of myrosinase binding protein and presence of genetic markers were also assayed. No correlation between degree of resistance and amount of sinigrin or other aliphatic glucosinolates was found. However, in time course experiments the glucosinolate profile of the L. maculans inoculated plants differed significantly from the water‐inoculated control plants in the amount of 4‐methoxy‐glucobrassicin observed. Five to eight days post‐inoculation an increased level of 4‐methoxy glucobrassicin, ranging from 30% to 47% on average, was found in the inoculated plants, whilst controls varied between 7.6% and 9.2%. This increase was seen both in susceptible and resistant material. Other changes observed could mainly be assigned as effects of wounding. Although inoculation with L. maculans elicited changes in the leaf indolyl glucosinolate profiles, in the material studied, these changes could not be correlated to resistance against the fungus.

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Isolation of Intact Glucosinolates by Column Chromatography and Determination of Their Purity

Cited by 17 publications

References 7 publications

Profiling glucosinolates in vegetative and reproductive tissues of four Brassica species of the U‐triangle for their biofumigation potential

Profiling glucosinolates in vegetative and reproductive tissues of four Brassica species of the U‐triangle for their biofumigation potential

Effects of Novel Processing Techniques on Glucosinolates and Membrane Associated Myrosinases in Broccoli

B‐genome derived resistance to Leptosphaeria maculans in near isogenic Brassica napus lines is independent of glucosinolate profile

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