Abstract | BACKGROUND: METHODS: LPCs are extracted from 3-mm DBS punch with methanol containing an isotopically labeled LPC as internal standard. This extract is transferred to a 96-well plate, evaporated and then reconstituted in mobile phase for flow injection analysis tandem mass spectrometry (FIA-MS/MS) in selected reaction monitoring mode for measurement of four different LPCs (C20, C22, C24, C26) and the internal standard (d4-C26-LPC). Analysis time is 1.5min per sample. RESULTS: The mean CVs from the intra- and inter-assay experiments for LPCs were 6.3-15.1% for C20-LPC, 4.4-18.6% for C22-LPC and 4.5-14.3% for C24-LPC. Limits of detection were determined for C20-LPC (LOD=0.03μg/mL), C22-LPC (0.03μg/mL), C24-LPC (0.03μg/mL) and C26-LPC (0.01μg/mL). Reference ranges were established from DBS of 130 newborns and 20 adults. Samples of patients with X-ALD (n=16), peroxisomal biogenesis disorders (n=8), and X-ALD carriers (n=12) were analyzed blindly and all were correctly identified. CONCLUSION: Analysis of LPC species by FIA-MS/MS is a fast, simple and reliable method to screen for X-ALD and other peroxisomal disorders in DBS. To maximize specificity, abnormal results can be verified by a 2nd tier assay using LC-MS/MS.
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Authors | Coleman T Turgeon, Ann B Moser, Lars Mørkrid, Mark J Magera, Dimitar K Gavrilov, Devin Oglesbee, Kimiyo Raymond, Piero Rinaldo, Dietrich Matern, Silvia Tortorelli |
Journal | Molecular genetics and metabolism
(Mol Genet Metab)
Vol. 114
Issue 1
Pg. 46-50
(Jan 2015)
ISSN: 1096-7206 [Electronic] United States |
PMID | 25481105
(Publication Type: Journal Article, Validation Study)
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Copyright | Copyright © 2014 Elsevier Inc. All rights reserved. |
Chemical References |
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Topics |
- Adrenoleukodystrophy
(blood)
- Adult
- Chromatography, Liquid
- Dried Blood Spot Testing
- High-Throughput Screening Assays
- Humans
- Infant, Newborn
- Limit of Detection
- Lysophosphatidylcholines
(blood)
- Neonatal Screening
(methods)
- Peroxisomal Disorders
(blood)
- Reference Values
- Tandem Mass Spectrometry
(methods)
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