Simultaneous Determination of Isothiocyanates, Indoles, and Oxazolidinethiones in Myrosinase Digests of Rapeseeds and Rapeseed Meal by HPLC

Abstract

HPLC has been used for the analysis of isothiocyanates, indoles, and oxazolidinethiones in rapeseeds and rapeseed meal. The samples were treated with myrosinase and the released hydrolysis products extracted with dichloromethane. The separation was performed on an RP-18 column using a gradient system with acetonitrile and water. Use of a programmable UV detector permitted the detection of the compounds at their absorption maxima of 210 and 240 nm, respectively. Response factors of eight standard compounds were calculated for 240 nm. The contents of glucosinolates calculated with the results of this method showed a significant linear correlation (r = 0.9995; P < 0.005) with the contents of glucosinolates evaluated with the results of the HPLC method of desulfoglucosinolates. Keywords: HPLC; indoles; isothiocyanates; oxazolidinethiones; rapeseed

Characterization of recombinant nitrile-specifier proteins (NSPs) of Arabidopsis thaliana: Dependency on Fe(II) ions and the effect of glucosinolate substrate and reaction conditions

“…Analysis was performed by HPLC-DAD (Agilent 1200 Series; Agilent Technologies, Santa Clara, USA) based on previously published protocols (Burow et al, 2008;Matthäus and Fiebig, 1996). The separation was carried out on a Supelcosil LC18 column (250 Â 2.1 mm, 5 lm) (Supelco, Bellefonte, PA, USA) with 10 lL injection volume and a flow rate of 0.5 mL.min À1 , at room temperature.…”

“…The following gradient program of solvent A (Milli-Q-filtered water) and solvent B (acetonitrile) was used: 0-5 min, 15% (v/v) B; 5-20 min, 15-75% (v/v) B; 20-25 min, 75-100% (v/v) B; 25-27 min, 100% (v/v) B; 27-27.10 min, 100-15% (v/v) B; 27.10-32.10 min, 15% (v/v) B. Products were detected with a diode array detector (DAD) at 240 nm (Matthäus and Fiebig, 1996). The data analysis was performed using Agilent Chemstation for LC 3D system software (Agilent Technologies, Santa Clara, USA).…”

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

“…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].…”

Determination of two intact glucosinolates in vegetables and Chinese herbs

“…While some glucosinolates and their breakdown products are found to have anti-nutritional effects in cattle [6], others have exhibited marginal anti-cancer activity [7,8,9]. Moreover, they have been shown to have activity-increasing effects on phase II detoxification enzymes that are related to cancer prevention [4].…”