Simultaneous quantitation of histamine and its major metabolite 1-methylhistamine in brain dialysates by using precolumn derivatization prior to HILIC-MS/MS analysis

Abstract

In nonclinical drug development targeting the central nervous system (CNS), the quantitative determination of extracellular brain concentrations of neurotransmitters is a key challenge. In some CNS disorders, the monitoring of the modified profile of neurotransmitter release such as that of histamine may explain the mechanism of action of the drug candidate. Microdialysis is a commonly used method for sampling extracellular levels of neurotransmitters/drug candidates in small laboratory animals. Detection and quantification of extracellular levels of neurotransmitters remain an analytical and technical challenge owing to the low concentrations of neurotransmitters collected, the small microdialysis sample size, and the high amount of inorganic salts. A precolumn derivatization strategy prior to hydrophilic interaction liquid chromatography (HILIC)-tandem mass spectrometry analysis is proposed to quantify histamine release after administration of a CNS research compound. Derivatization using propionic anhydride dissolved in organic solvent combined with the HILIC approach effectively eliminated three time-consuming steps, organic layer transfer, dry down, and reconstitution, all of which are required by traditional reversed-phase liquid chromatography. The formation of propionylated amides, performed under mild conditions, required no further sample cleanup. After a dual microdialysis probe implantation into the prefrontal cortex (neurotransmitters) and in the inferior vena cava of rat (drug candidate), microdialysate fractions were collected every 15 min for 8 h and stored frozen at -20 °C until analysis. The method was validated using 10 μL microdialysate, achieving low limits of quantitation of 83.4 and 84.5 pg.mL(-1) for histamine and 1-methylhistamine, respectively. These limits were suitable to assess kinetic release of neurotransmitters and are compatible with those obtained by microdialysis sampling. This method provided the required selectivity, sensitivity, accuracy, and precision to assess release kinetics of histamine and 1-methylhistamine in several hundred rat brain microdialysates after intravenous infusion of CNS drug candidates.

Histamine Stimulates Neurogenesis in the Rodent Subventricular Zone

“…Importantly, the real histamine cumulative released from microparticles was far smaller (×1,000 less) than the initial histamine concentrations tested for the single‐pulse experiments (100 μM). Several studies reported that in physiologic conditions histamine is present at nanomolar concentrations in the cerebrospinal fluid and brain parenchyma of humans and rodents [38–44]. Importantly, circulating levels of histamine and histaminergic innervations are increased following brain injury and degeneration or infection [39, 45‐49].…”

New insights into the role of histamine in subventricular zone-olfactory bulb neurogenesis

“…Histaminergic neurons, present in the tuberomammillary nucleus, project numerous ramifications throughout the entire adult brain, allowing histamine to be involved in a broad range of physiological functions, such as sleep-wake control, emotions, learning and memory (Panula and Nuutinen, 2013). Histamine is found at nanomolar levels in the healthy brain (Soya et al, 2008; Croyal et al, 2011; Bourgogne et al, 2012). However, several brain pathological conditions may be associated with an increased degranulation of mast cells in the choroid plexus , leading to a massive release of histamine in the CSF and the consequent increase of the blood brain barrier (BBB) permeability.…”

Development and validation of a liquid-chromatography tandem mass spectrometry method to determine in vitro and in vivo histamine release

“…As reversed-phase chromatographic analysis is a challenge due to the high polarity of histamine, HILIC chromatography was employed for enhanced retention of histamine. The LC-MS/MS assay validated in the present study was modified from Bourgogne et al, 2012 [22]. Significant matrix effect was observed from the cell culture media with cell lysates with the published LC gradient.…”

“…Because HILIC methods use high concentration of acetonitrile as the mobile phase, it is possible to directly inject strong solvents onto the column after appropriate sample preparation without introducing peak distortion from solvent effects [20,21]. Using a HILIC based approach, Bourgogne et al were able to detect derivatized histamine and its major metabolite from brain dialysates [22] The primary objective of the present study was to develop and validate a rapid and sensitive HILIC-MS/MS method to quantify chemically induced-histamine release in in vitro and in vivo models without the derivatization and dialysis step. In vitro release of histamine was induced by treating isolated rat peritoneal mast cells with Compound 48/80 or Teicoplanin, chemicals widely known to directly degranulate mast cells and cause histamine release [23,24].…”