High-resolution mass spectrometry assay for quantifying ceramides and dihydroceramides in the cerebrospinal fluid from patients who experienced intracranial infection after craniotomy

Abstract

Ceramides (CERs) and dihydroceramides (dhCERs) are biologically active lipids involved in cell proliferation, differentiation, and apoptosis, as well as associated with infectious diseases. The concentration of CERs and dhCERs in the cerebrospinal fluid (CSF) could potentially allow the distinction of patients with intracranial infection (ICI) after craniotomy from controls, but the quantification is limited by their ultralow concentrations. Therefore, a novel high performance liquid chromatography-triple-quadrupole/time-of-flight mass spectrometry (HPLC-QTOF-MS) with Sequential Window Acquisition of All Theoretical Fragment Ion Spectra (SWATH) was applied to measure CERs and dhCERs in CSF, since it possesses a higher sensitivity (LLOQ = 0.1 pmol/mL) than the multiple reaction monitoring (MRM) acquisition carried on triple quadrupole (QqQ) MS. This method was validated and applied to CSF samples from patients who experienced postoperative ICI (63 patients) and controls who did not experience it after surgery (62 patients). This assay was linear over the measuring range 0.1-100 pmol/mL for these CER and dhCER species with good accuracy and precision. Elevated CERs and dhCERs were observed in CSF from patients who experienced postoperative ICI. CER 16:0 was found with a clinical sensitivity of 93.65 % and specificity of 87.1 % in distinguishing the 63 patients with ICI from the 62 controls. Therefore, this method could be applied in the detection of CERs and dhCERs in CSF, which were correlated with the presence of ICI.

Keywords: Ceramide; Dihydroceramide; HRMS; Intracranial infection after craniotomy; SWATH.