Rapid quantitation of globotriaosylceramide in human plasma and urine: a potential application for monitoring enzyme replacement therapy in Anderson-Fabry disease.
A method for measuring globotriaosylceramide (Gb3, or GL3) levels in plasma and urine of humans affected by Anderson-Fabry disease has been developed. The analyses are performed using flow injection analysis-electrospray ionization-tandem mass spectrometry (FIA-ESI-MS/MS). The method is rapid, sensitive and hence suitable for high-throughput analyses, requiring only a simple 50-fold dilution for the preparation of plasma and urine samples. The detection of the analytes of interest was achieved using a triple quadrupole instrument operating in the multiple reaction monitoring mode. The linearity of the calibration standard responses, the intra- and inter-assay precision, the accuracy and the detection limit of the method were evaluated. The proposed method allows a rapid and accurate assessment of globotriaosylceramide in biological samples. Data obtained from healthy volunteers and Anderson-Fabry affected subjects suggest a potential role for this technique in monitoring the effectiveness of Anderson-Fabry disease therapy. The results obtained in two actual cases treated with enzyme replacement therapy are reported and discussed.
Centro Interdipartimentale di Spettrometria di Massa, Università di Firenze, Viale G. Pieraccini 6, 50139 Firenze, Italy.
Francesca
F
Boscaro
Giuseppe
G
Pieraccini
Giancarlo
G
la Marca
Gianluca
G
Bartolucci
Cristina
C
Luceri
Francesca
F
Luceri
Gloriano
G
Moneti
D000795:Fabry Disease / Q000097:blood / Q000188:drug therapy* / Q000652:urine; D005260:Female; D006801:Humans; D008297:Male; D011994:Recombinant Proteins / Q000627:therapeutic use; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D013696:Temperature; D014281:Trihexosylceramides / Q000032:analysis* / Q000097:blood / Q000652:urine; D000519:alpha-Galactosidase / Q000627:therapeutic use*
D011994:Recombinant Proteins; D014281:Trihexosylceramides; C018549:globotriaosylceramide; D000519:alpha-Galactosidase