A new method of analysis based on normal phase supercritical fluid chromatography (SFC) hasbeen developed for investigation of ascorbigens [2-C-(indol-3-ylmethyl)-
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-
L-xylo-3-hexulofuranosonicacid
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-lactone derivatives]. This method has been adapted to preparative isolation and quantitativedeterminations of individual ascorbigens comprising ascorbigen, neoascorbigen, and 4-methoxyascorbigen. The structures of these compounds have been revealed from 1D (
1H,
13C) and 2D (COSY,HMQC, HMBC) NMR experiments. The developed SFC method had an acceptable linearity for theascorbigens with correlation coefficients (
R2) > 0.9995 (
n = 10) in the range of 0.13-4.9 nmol injected,detection limits were below 13 pmol, retention time stabilities were excellent, and relative responsefactors have been determined. The SFC method has been used for determination of ascorbigensproduced during autolysis of indol-3-ylmethylglucosinolates in various
Brassica vegetables andrapeseed seedlings. Generally, 30-60% of the indol-3-ylmethylglucosinolates in the plants weretransformed into ascorbigens, with the concentration in autolysates varying from 0.51 ± 0.002 to3.72 ± 0.21
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mol/g of dry weight (DW) for ascorbigen, from 0.05 ± 0.01 to 2.42 ± 0.23
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mol/g of
DWfor neoascorbigen, and from 0.03 ± 0.002 to 0.84 ± 0.07
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mol/g of DW for 4-methoxyascorbigen.Keywords: Brassicaceae; Brassica; glucosinolate; ascorbigens; supercritical fluid chromatography