Quantitation and Standardization of Lipid Internal Standards for Mass Spectroscopy

Quantitative analysis of sphingolipids for lipidomics using triple quadrupole and quadrupole linear ion trap mass spectrometers

“…The LIPID MAPS™ internal standard cocktail (initially Sphingolipid Mix I, catalog number LM-6002; later replaced by Sphingolipid Mix II, catalog number LM-6005 as explained under "Results") was provided by Avanti Polar Lipids (Alabaster, AL) in sealed ampules and certifi ed ( 35 ) to be over 95% pure and within 10% of the specifi ed amount (25 µM). It was composed of the 17-carbon chain length sphingoid base analogs C17-sphingosine, (2S,3R,4E)-2-aminoheptadec-4-ene-1,3-diol (d17:1-So) 3 ; C17-sphinganine, (2S,3R)-2-aminoheptadecane-1,3-diol (d17:0-Sa); C17-sphingosine 1-phosphate, heptadecasphing-4-enine-1-phosphate (d17:1-So1P); and C17-sphinganine 1-phosphate, heptadecasphinganine-1-phosphate (d17:0-Sa1P); and the C12-fatty acid analogs of the more complex sphingolipids C12-Cer, N-(dodecanoyl)-sphing-4-enine (d18:1/C12:0); C12-Cer 1-phosphate, N-(dodecanoyl)-sphing-4-enine-1-phosphate (d18:1/C12:0-Cer1P); C12-sphingomyelin, N-(dodecanoyl)-sphing-4-enine-1-phosphocholine (d18:1/C12:0-SM); C12-glucosylceramide, N-(dodecanoyl)-1-␤ -glucosyl-sphing-4-eine (d18:1/C12:0-GlcCer); and C12-lactosylceramide, N-(dodecanoyl)1-␤ -lactosyl-sphing-4-eine (d18:1/ The elution times for these analytes ( Fig.…”

Analysis of mammalian sphingolipids by liquid chromatography tandem mass spectrometry (LC-MS/MS) and tissue imaging mass spectrometry (TIMS)

“…A more practical approach has been introduced to identify internal standards that are similar in structure and physical properties, as well as ionization and fragmentation characteristics for the categories of compounds under investigation. To this end, an internal standard cocktail has been developed for the LIPID MAPS Consortium [5, 6] (which is commercially available from Avanti Polar Lipids) that contains uncommon chain-length sphingoid bases (C17) for sphingosine (So), sphinganine (Sa) and their 1-phosphates (S1P and Sa1P) and C12:0 fatty acid analogs of Cer, ceramide 1-phosphate (Cer1P), SM, and mono- and dihexosylCer (HexCer and diHexCer). The types of analytes can be expanded by supplementation with additional internal standards.…”

Lipidomics joins the omics evolution

“…Combined with the use of internal standards, these techniques allow for the identification and quantification of the lipids known to be present, but do not, by themselves, lead to the finding of novel or unexpected molecular species. Ejsing et al (5) made critical use of multiple lipid standards obtained from Avanti Polar Lipids which had been developed under LIPID MAPS auspices (14), but they also had to develop their own standards for some specific yeast phytosphingolipids. The yeast lipidome is characterized by the usual TAG, DAG, and phospholipid species found in mammalian cells, but in addition has unusual phytosphingolipid species in which the long-chain base and the fatty acyl amide moieties are characterized by hydroxylated species (e.g., phytoceramides) and an abundance of the sterol ergosterol rather than the mammalian cholesterol.…”