TY - JOUR
T1 - Ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) for determination of GHB, precursors and metabolites in different specimens
T2 - Application to clinical and forensic cases
AU - Busardò, Francesco Paolo
AU - Kyriakou, Chrystalla
AU - Marchei, Emilia
AU - Pacifici, Roberta
AU - Pedersen, Daniel Sejer
AU - Pichini, Simona
N1 - Copyright © 2017 Elsevier B.V. All rights reserved.
PY - 2017/4/15
Y1 - 2017/4/15
N2 - Gamma-hydroxybutyric acid (GHB) acts as a precursor and metabolite of the inhibitory central nervous system (CNS) neurotransmitter gamma-aminobutyric acid (GABA). Sodium salt of GHB has been used as a medication for narcolepsy and alcohol withdrawal. Moreover, GHB and its precursor gamma-butyrolactone (GBL), are illegal recreational drugs of abuse. A procedure based on ultra-high-performance liquid chromatography tandem mass spectrometry has been developed and validated in plasma, urine, cerebrospinal fluid and hair for acute and chronic exposure to GHB and in seized preparations coming from black market. In biological matrices, GHB was investigated together with its glucuronide (GHB-Gluc) as a potential marker of exposure, GABA as endogenous precursor and metabolite and GBL as eventual exogenous precursor. GBL was sought together with GHB in illegal preparations. Chromatographic separation was achieved at ambient temperature using a reverse-phase column and an isocratic elution with two solvents: 0.1% formic acid in water and pure methanol. Multiple reaction monitoring (MRM) was used. The method was linear for all analytes under investigation from limit of quantification (LOQ) to 500μgmL-1 plasma, urine and cerebrospinal fluid, from LOQ to 100ngmg-1 hair and from LOQ to 10mgmL-1 illicit preparations with good correlation coefficients (r2=0.99) for all substances. Recovery of analytes under investigation was always higher than 75% and intra-assay and inter-assay precision and accuracy were always better than 15%. The validated method was then successfully applied to real specimens from either forensic (one post-mortem urine sample taken from a GHB fatal intoxication case) or clinical cases (cerebrospinal fluid, plasma and hair samples collected from narcoleptic patients under sodium oxybate treatment). Finally, illicit preparations, seized by police forces were also checked for GHB amount and eventual presence of prodrug GBL.
AB - Gamma-hydroxybutyric acid (GHB) acts as a precursor and metabolite of the inhibitory central nervous system (CNS) neurotransmitter gamma-aminobutyric acid (GABA). Sodium salt of GHB has been used as a medication for narcolepsy and alcohol withdrawal. Moreover, GHB and its precursor gamma-butyrolactone (GBL), are illegal recreational drugs of abuse. A procedure based on ultra-high-performance liquid chromatography tandem mass spectrometry has been developed and validated in plasma, urine, cerebrospinal fluid and hair for acute and chronic exposure to GHB and in seized preparations coming from black market. In biological matrices, GHB was investigated together with its glucuronide (GHB-Gluc) as a potential marker of exposure, GABA as endogenous precursor and metabolite and GBL as eventual exogenous precursor. GBL was sought together with GHB in illegal preparations. Chromatographic separation was achieved at ambient temperature using a reverse-phase column and an isocratic elution with two solvents: 0.1% formic acid in water and pure methanol. Multiple reaction monitoring (MRM) was used. The method was linear for all analytes under investigation from limit of quantification (LOQ) to 500μgmL-1 plasma, urine and cerebrospinal fluid, from LOQ to 100ngmg-1 hair and from LOQ to 10mgmL-1 illicit preparations with good correlation coefficients (r2=0.99) for all substances. Recovery of analytes under investigation was always higher than 75% and intra-assay and inter-assay precision and accuracy were always better than 15%. The validated method was then successfully applied to real specimens from either forensic (one post-mortem urine sample taken from a GHB fatal intoxication case) or clinical cases (cerebrospinal fluid, plasma and hair samples collected from narcoleptic patients under sodium oxybate treatment). Finally, illicit preparations, seized by police forces were also checked for GHB amount and eventual presence of prodrug GBL.
KW - 4-Butyrolactone
KW - Cerebrospinal Fluid
KW - Chromatography, High Pressure Liquid
KW - Forensic Sciences
KW - Hair
KW - Humans
KW - Hydroxybutyrates
KW - Male
KW - Methanol
KW - Middle Aged
KW - Plasma
KW - Sodium Oxybate
KW - Street Drugs
KW - Tandem Mass Spectrometry
KW - gamma-Aminobutyric Acid
KW - Journal Article
U2 - 10.1016/j.jpba.2017.01.022
DO - 10.1016/j.jpba.2017.01.022
M3 - Journal article
C2 - 28110168
SN - 0731-7085
VL - 137
SP - 123
EP - 131
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
ER -