Abstract
The orthogonal heart-cutting liquid chromatography (LC) modes coupled to high-resolution tandem mass spectrometry (HRMS/MS) provide a number of possibilities to enhance selectivity and sensitivity for the determination of targeted compounds in complex biological matricies. Here we report the development of a new fast 2D-LC-(HRMS/MS) method and its successful application for quantitative determination of the level of plasma and brain N,N-dimethyltriptamine (DMT) using α-methyltryptamine (AMT) as internal standard in an experimental model of cerebral ischemia/reperfusion using DMT administration. The 2D-LC separation was carried out by a combination of hydrophilic interaction liquid chromatography (HILIC) in the first dimension followed by second-dimensional reversed-phase (RP) chromatography within a total run time of 10 min. The enrichment of HILIC effluent of interest containing DMT was performed using a C18 trapping column. During method development several parameters of sample preparation procedures, chromatographic separation and mass spectrometric detection were optimised to achieve high DMT recovery (plasma: 90 %, brain: 88 %) and sensitivity (plasma: 0.108 ng/mL of LOD, brain: 0.212 ng/g of LOD) applying targeted analytical method with strict LC and HRMSMS confirmatory criteria.Concerning rat plasma sample, the concentration of DMT before hypoxia (49.3–114.3 ng/mL plasma) was generally higher than that after hypoxia (10.6–96.1 ng/mL plasma). After treatment, the concentration of DMT in brain was elevated up to the range of 2–6.1 ng/g.Overall, our analytical approach is suitable to detect and confirm the presence of DMT administered to experimental animals with therapeutic purpose in a reliable manner.
Highlights
DMT is an endogenous hallucinogen and trace amine found in various tissues such as the brain, pineal gland and lung, and body fluids such as urine, cerebrospinal fluid and blood plasma in mammals [1,2,3,4]
DMT is produced by somatic cells under physiological conditions [1,3], and its concentration has been found to increase in response to pathophysiological homeostatic challenges [1,3]
N,N-Dimethyltryptamine (DMT) standard with purity >98.5 % was obtain from Lipomed AG (Arlesheim, Switzerland). ␣Methyltryptamine (AMT) with purity >99 % was kindly provided by the Department of Forensic Medicine, Faculty of Medicine, University of Szeged, Hungary, and used as internal standard (IS)
Summary
DMT is an endogenous hallucinogen and trace amine found in various tissues such as the brain, pineal gland and lung, and body fluids such as urine, cerebrospinal fluid and blood plasma in mammals [1,2,3,4]. DMT is produced by somatic cells under physiological conditions [1,3], and its concentration has been found to increase in response to pathophysiological homeostatic challenges (e.g. hypoxia or oxidative stress) [1,3]. DMT has been implicated in neuroprotection at high concentration, either synthesised in the nervous tissue in response to hypoxia or oxidative stress, or originating from the lung and reaching the brain via the blood stream [1,6]. DMT administered at supraphysiological concentration to experimental rodents resulted in a smaller cerebral infarct size [7] and promoted cell survival in a
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