Rapid analysis of endogenous LVV-hemorphin-7 in cerebrospinal fluid by size-exclusion chromatography and electrospray ionization mass spectrometry

Abstract

LVV-hemorphin-7 is a peptide with opioid-like activity which is recognized by μ and σ receptors. The sequence, derived from β-, γ-, δ- or ϵ- chains of human hemoglobin has been found in cerebrospinal fluid (CSF) of patients suffering from cerebrovascular bleedings, but not in CSF from healthy individuals. Procedures for isolation and identification of neuropeptides in body fluids and tissue extracts are often laborious and time-consuming. Additionally, extraction yield tends to be low after several chromatographic steps. In this study, we developed a rapid technique to analyze LVV-hemorphin-7 in CSF fluid from a patient with cerebrovascular bleedings using a combination of size-exclusion chromatography and electrospray ionization mass spectrometry. The methodology described utilizes small quantities of CSF (0.3–0.5 ml) and can be completed within a few hours. The proposed strategy opens possibilities for direct measurement of endogenous peptides in small volumes of body fluids.