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KMID : 0370220070510020096
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Yakhak Hoeji
2007 Volume.51 No. 2 p.96 ~ p.102
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Method Development for the Profiling Analysis of Urine Globotriaosylceramide (Gb3) for the Screening of Fabry Disease by Tandem Mass Spectrometry
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Yoon Hye-Ran
Kwon Young-Joo
Jeong Choon-Sik
Lee Yong-Soo
Kang Seong-Woo
Cho Kyunghee
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Abstract
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Measurement of globotriaosylceramide (Gb3, ceramide trihexoside) in urine has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme {alpha}-galactosidase A ({alpha}-Gal A). The lack of {alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. A simple, rapid, and highly sensitive analytical method for Gb3 in urine was developed without labor-extensive pre-treatment by electrospray ionization MS/MS (ESI-MS/MS). Only simple 5-fold dilution of urine is necessary for the extraction and isolation of Gb3 in urine. Gb3 in diluted urine was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Eight isoforms of Gb3 were completely resolved from urine matrix. C24:0 Gb3 occupied 50% of total Gb3 as a major component in urine. Linear relationship for Gb3 isoforms was found in the range of 0.005{sim}5.0 {mu}g/ml. The limit of detection (S/N=5) was 0.005 {mu}g/ml and limit of quantification was 0.05 {mu}g/ml for C24:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9598 to 0.9975. This method could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.
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KEYWORD
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quantification, globotriaosylceramide (Gb3), MS/MS, fabry disease, screening
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