2003
DOI: 10.1016/s0021-9673(03)00607-1
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Separation and purification of glucosinolates from crude plant homogenates by high-speed counter-current chromatography

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Cited by 80 publications
(41 citation statements)
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“…All reagents used were analytic or high-performance liquid chromatography (HPLC) grade. Glucosinolate standards used for HPLC were isolated and identified according to published methods (30)(31)(32).…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…All reagents used were analytic or high-performance liquid chromatography (HPLC) grade. Glucosinolate standards used for HPLC were isolated and identified according to published methods (30)(31)(32).…”
Section: Methodsmentioning
confidence: 99%
“…The hot aqueous extracts were subsequently analyzed by HPLC according to methods developed for separation of intact glucosinolates (30). Confirmation of glucosinolate identities was done by comparison of retention times with previously obtained glucosinolate standards and by mass spectrometry (31,32). Infusions were also analyzed following digestion with added myrosinase and ascorbic acid as described by Shapiro et al (33).…”
Section: Methodsmentioning
confidence: 99%
“…When broccoli sprouts are chipped, mechanically damaged or attacked by pest, glucosinolates directly bond with myrosinase and are mainly converted into isothiocyanates (ITCs), such as sulforaphane, erucin and iberin (Grubb and Abel 2006;Latté et al 2011). Myrosinase-catalysed hydrolysis of glucosinolates initially involves cleavage of the thioglucoside linkage, yielding D-glucose and an unstable thiohydroximate-O-sulfonate that is spontaneously rearranged, which results in production of sulfate and a wide range of possible reaction products (Fahey et al 2003;Angelino and Jeffery 2014). Sulforaphane (4-methylsulfinylbutyl isothiocyanate) is a naturally occurring, sulfur-containing, ITCs with excellent capacity to induce phase 2 enzymes, which are formed from glucosinolate (Nakagawa et al 2006;Yeh and Yen 2009).…”
Section: Introductionmentioning
confidence: 99%
“…We have used this method to identify and quantify glucosinolates in broccoli (Brassica oleracea var. italica), as well as in other plants [13][14][15][16][17][18][19][20]. The HILIC columns utilized (polyhydroxyethyl aspartamide) have short life-spans (only a few hundred injections) compared to C18 columns, and the mobile phase required to effect such separation does not well tolerate slight changes in composition to resolve certain pairs or groups of co-eluting glucosinolates over others.…”
Section: Introductionmentioning
confidence: 99%
“…For development and scale-up of a novel (counter-current chromatographic) method for purifying GR (glucoraphanin) from broccoli seeds [14,22], we used HILIC to evaluate the degree of separation of GR from its most persistent "contaminant", glucoriberin (GI). Since these separations were being performed both in Europe and in the USA, we evaluated alternative HILIC column chemistries (that were readily available from companies with worldwide distribution networks), as well as altering HPLC variables such as pH, organic acids and water activity.…”
Section: Introductionmentioning
confidence: 99%