Supercritical fluid chromatography as a method of analysis for the determination of 4-hydroxybenzylglucosinolate degradation products.
In the present study analytical and preparative supercritical fluid chromatography (SFC) were used for investigation of myrosinase catalysed degradation of 4-hydroxybenzylglucosinolate (sinalbin). Sinalbin occurs as a major glucosinolate in seeds of Sinapis alba L., in various mustards and other food products. The degradation products were identified and quantified by analysis based on a developed SFC method using a bare silica column. Determinations comprised transformation products of sinalbin, produced both during degradation of isolated sinalbin, and during autolysis of meal from S. alba seeds. The conditions in the developed SFC method were used as basis for the preparative SFC procedure applied for isolation of the components prior to their identification by nuclear magnetic resonance (NMR) spectroscopy. Myrosinase catalysed sinalbin hydrolysis resulted in the reactive 4-hydroxybenzyl isothiocyanate as an initial product at pH values from 3.5 to 7.5 whereas 4-hydroxybenzyl cyanide was one of the major products at low pH values. 4-Hydroxybenzyl isothiocyanate was found to disappear from the aqueous reaction mixtures in a few hours, as it reacted easily with available nucleophilic reagents. 4-Hydroxybenzyl alcohol was found as the product from reaction with water, and with ascorbic acid, 4-hydroxybenzylascorbigen was produced.
Buskov
S
S
Chemistry Department, Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871, Frederiksberg C, Denmark.
Hasselstrøm
J
J
Olsen
C E
CE
Sørensen
H
H
Sørensen
J C
JC
Sørensen
S
S
D000097:Acetonitriles / Q000302:isolation & purification; D001205:Ascorbic Acid / Q000302:isolation & purification; D001592:Benzyl Alcohols / Q000302:isolation & purification; D002794:Choline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification; D002845:Chromatography / Q000295:instrumentation* / Q000379:methods*; D005961:Glucosinolates / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D017879:Isothiocyanates / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D009149:Mustard Plant / Q000737:chemistry; D010936:Plant Extracts / Q000302:isolation & purification; D010946:Plants, Medicinal; D012639:Seeds / Q000737:chemistry; D013997:Time Factors; D014269:Trifluoroacetic Acid / Q000737:chemistry
C097342:4-hydroxybenzylglucosinolate; D000097:Acetonitriles; D001592:Benzyl Alcohols; D005961:Glucosinolates; D017879:Isothiocyanates; D010936:Plant Extracts; C026914:4-hydroxybenzylcyanide; C018966:4-hydroxybenzyl alcohol; C031364:4-hydroxybenzyl isothiocyanate; D014269:Trifluoroacetic Acid; D002794:Choline; D001205:Ascorbic Acid; C003313:sinalbin