Nano-LC–MS/MS for the quantitation of ceramides in mice cerebrospinal fluid using minimal sample volume

A new nano-liquid chromatography–ESI–MS/MS method has been developed for the sensitive quantitation of C8:0, C16:0, C18:0, C18:1, C20:0, C24:1 and C24:0 ceramide in cerebrospinal fluid of mice using minimal sample volume. Volumes of 2µL CSF were undertaken a simple, fast extraction procedure involvi...

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Vydáno v:Talanta (Oxford) Ročník 116; s. 912 - 918
Hlavní autoři: Thomas, D., Eberle, M., Schiffmann, S., Zhang, D.D., Geisslinger, G., Ferreirós, N.
Médium: Journal Article
Jazyk:angličtina
Vydáno: Netherlands Elsevier B.V 15.11.2013
Témata:
Animals
Calibration
Ceramide
ceramides
Ceramides - cerebrospinal fluid
Ceramides - classification
Cerebrospinal fluid
Chromatography, Liquid
Female
Limit of Detection
methanol
Methanol - chemistry
Mice
Mice, Inbred C57BL
Nano-LC–MS/MS
Nanotechnology - instrumentation
Nanotechnology - methods
Protein Denaturation
Reference Standards
Reproducibility of Results
reversed-phase liquid chromatography
Specimen Handling
spectrometers
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Ceramide
Nano-LC–MS/MS
Cerebrospinal fluid
ISSN:0039-9140, 1873-3573, 1873-3573
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Shrnutí:A new nano-liquid chromatography–ESI–MS/MS method has been developed for the sensitive quantitation of C8:0, C16:0, C18:0, C18:1, C20:0, C24:1 and C24:0 ceramide in cerebrospinal fluid of mice using minimal sample volume. Volumes of 2µL CSF were undertaken a simple, fast extraction procedure involving protein precipitation with methanol and dilution. Ceramides were separated by nano-liquid chromatography with a reversed phase C8 column and detected with a triple quadrupole mass spectrometer. C17:0 ceramide was used as internal standard. The method has been validated in terms of linearity, lower limit of quantitation, precision, accuracy and autosampler stability. Calibration curves covered a range of 2.25–120pg/µL for most ceramides (7.5–120pg/µL for C24:0 ceramide). The lower limits of quantitation determined for C8:0, C16:0, C18:1, C18:0, C20:0 and C24:1 ceramide were 0.225pg on column (2.25pg/µL) and that for C24:0 ceramide 0.750pg on column (7.5pg/µL). Intra- and interday precision and accuracy values, expressed as relative standard deviation and relative error, respectively, were lower than 15% in all cases. Autosampler stability for calibration standards and CSF samples was proven for at least 24h for all analytes. The suitability of the method has been demonstrated by quantifying the analytes, except the non-endogenous C8:0 ceramide, in cerebrospinal fluid samples of 12 mice. Calculated concentrations ranged from 3 to 120pg/µL in cerebrospinal fluid for all analytes, except for C24:0 ceramide, which could not be detected in any of the analyzed samples. •A very high sensitive method for ceramides in cerebrospinal fluid was developed.•Only 2µL sample volume was required.•Validation according to the FDA guidelines was performed.•First approach using nano-LC–MS/MS for quantitation of ceramides.
Bibliografie:http://dx.doi.org/10.1016/j.talanta.2013.07.057
ObjectType-Article-1
SourceType-Scholarly Journals-1
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content type line 23
ISSN:0039-9140
1873-3573
1873-3573
DOI:10.1016/j.talanta.2013.07.057