Determination of sphingoid bases from hydrolyzed glucosylceramide in rice and wheat by online post-column high-performance liquid chromatography with O-phthalaldehyde derivatization.
J Oleo Sci
; 61(12): 681-8, 2012.
Article
em En
| MEDLINE
| ID: mdl-23196868
ABSTRACT
We developed a determination method for sphingoid bases using online post-column high-performance liquid chromatography (HPLC) with O-phthalaldehyde (OPA) derivatization. Good separation was achieved using a reversed-phase column and eluting with 50% acetonitrile containing formic acid and heptafluorobutyric acid. Using these conditions, an excellent linearity (R² > 0.999) was achieved using standard solutions of sphinganine (d180), sphingosine (d181(4t)), 4-hydroxy-sphinganine (t180), glucosylsphingosine (glc-d181(4t)), and galactosylsphingosine (gal-d181(4t)). Plant glucosylceramides were hydrolyzed with 1 M aqueous HCl in methanol for 18 h at 90°C, followed by extraction of sphingoid bases with diethyl ether in preparation for analysis using the proposed HPLC conditions. The glc-d181(4t) standard was also hydrolyzed and analyzed by HPLC using the same procedure, and the d181(4t) peak obtained from the hydrolyzed glc-d181(4t) standard was used as a reference for calculation. We also confirmed the applicability of this method to the analysis of sphingoid bases in rice and wheat, obtaining relative standard deviations of 8.0% for rice and 4.6% for wheat. The recoveries of spiked rice and wheat samples were 104% and 106%, respectively. Our proposed method enables the straightforward determination of sphingoid bases without expensive facilities, employing fluorescence detection of OPA derivatives.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Oryza
/
Esfingosina
/
Triticum
/
O-Ftalaldeído
/
Internet
/
Glucosilceramidas
Idioma:
En
Revista:
J Oleo Sci
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Japão