Chapter title |
A Simple, High-Throughput Method for Analysis of Ceramide, Glucosylceramide, and Ceramide Trihexoside in Dried Blood Spots by LC/MS/MS
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Chapter number | 28 |
Book title |
Clinical Applications of Mass Spectrometry in Biomolecular Analysis
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Published in |
Methods in molecular biology, January 2016
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DOI | 10.1007/978-1-4939-3182-8_28 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3181-1, 978-1-4939-3182-8
|
Authors |
Wei-Lien Chuang, Joshua Pacheco, Kate Zhang |
Abstract |
A unique monophasic extraction system coupled with LC/MS/MS to reduce matrix effects for sphingolipid analysis was developed. A solvent mixture of methanol, acetonitrile, and water was identified to simultaneously extract multiple sphingolipids with broad polarity range. To reduce matrix effects, the targeted sphingolipids were analyzed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The extraction solvent was used as an isocratic mobile phase in chromatographic separation to eliminate solvent exchange steps and enable high-throughput multiple lipid assay. The assay is linear for ceramide from 0.6 to 9 μg/mL with bias <15 %. The intra-assay coefficient of variation is less than 10 % for concentrations from 1.2 to 9 μg/mL, and less than 25 % for concentrations below 1.2 μg/mL. For glucosylceramide and ceramide trihexoside the linear range is 0.05-3 μg/mL with biases <10 % and <20 %, respectively. The intra-assay coefficient of variation for these analytes is less than 10 % at concentrations from 0.4 to 3 μg/mL, and less than 25 % for concentrations below 0.4 μg/mL. |
Mendeley readers
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Demographic breakdown
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Other | 3 | 27% |
Student > Doctoral Student | 2 | 18% |
Student > Bachelor | 1 | 9% |
Student > Ph. D. Student | 1 | 9% |
Student > Postgraduate | 1 | 9% |
Other | 0 | 0% |
Unknown | 3 | 27% |
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Unknown | 4 | 36% |