Long-term stability of GHB in post-mortem samples and samples from living persons, stored at −20°C, using fluoride preservatives

Fjeld, Bente; Burns, Margrete Larsen; Karinen, Ritva; Larssen, Bente; Smith-Kielland, Anne; Vindenes, Vigdis

doi:10.1016/j.forsciint.2012.04.033

Cited by 19 publications

(15 citation statements)

References 23 publications

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“…Blood samples stabilized with NaF (n = 27 living individuals) were stored at −20°C and measured for GHB. Long‐term stability could be demonstrated over a period of 7 years, and the deviation to the primary concentrations ranged between −32.4% and 21.0% 37 . A good stability could also be demonstrated for serum samples without stabilization.…”

Section: Resultsmentioning

confidence: 86%

Phase I metabolites (organic acids) of gamma‐hydroxybutyric acid–validated quantification using GC–MS and description of endogenous concentration ranges

Jarsiah

Roehrich

Wyczynski³

et al. 2020

Drug Testing and Analysis

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Gamma‐hydroxybutyric acid (GHB) is a sedative drug used in drug‐facilitated crimes. Its detection window is very short. GHB undergoes intensive phase I metabolism to organic acids (glycolic acid, succinic acid, dihydroxybutyric acids). These could be potential analytical targets to broaden the detection window. The aim of the present study was to enable the detection of endogenous levels of these metabolites in biological samples (blood and urine). A gas chromatographic–mass spectrometric method using liquid–liquid extraction and derivatization with N‐methyl‐N‐tri‐methylsilyltrifluoracetamide was developed for the quantification. Validation results were consistent with international guidelines, and the method was able to quantify endogenous levels of the substances in both urine and blood. Endogenous concentrations were shown to be <0.03–4.92 mg/L for glycolic acid, <0.03–1.28 mg/L for GHB, <0.28–18.1 mg/L for succinic acid, <0.12–1.38 mg/L for 2,4‐dihydroxybutyric acid, and <0.13–2.59 mg/L for 3,4‐dihydroxybutyric acid in serum samples of 101 volunteers. Urinary endogenous concentrations were shown to be 1.30–400 mg/L for glycolic acid, <0.03–1.94 mg/L for GHB, 1.17–2.73 mg/L for succinic acid, 0.72–26.2 mg/L for 2,4‐dihydroxybutyric acid, and 1.88–122 mg/L for 3,4‐dihydroxybutyric acid in urine samples of 132 volunteers. These endogenous concentrations represent a basis to which concentrations after the intake of GHB can be compared to in order to prove the intake of this substance.

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Section: Resultsmentioning

confidence: 86%

Phase I metabolites (organic acids) of gamma‐hydroxybutyric acid–validated quantification using GC–MS and description of endogenous concentration ranges

Jarsiah

Roehrich

Wyczynski³

et al. 2020

Drug Testing and Analysis

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“…The presence of fluoride is important and helps to stabilize GHB concentrations, such as whenever there is a long delay between voiding and performing the analysis. For long-term storage, the urine specimens should be stored frozen [144].…”

Section: Urinementioning

confidence: 99%

Interpreting γ-hydroxybutyrate concentrations for clinical and forensic purposes

Busardò

Jones

2018

Clinical Toxicology

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Introduction: c-Hydroxybutyric acid is an endogenous substance, a therapeutic agent, and a recreational drug of abuse. This psychoactive substance acts as a depressant of the central nervous system and is commonly encountered in clinical and forensic practice, including impaired drivers, poisoned patients, and drug-related intoxication deaths. Objective: The aim of this review is to assist clinical and forensic practitioners with the interpretation of c-hydroxybutyric acid concentrations in blood, urine, and alternative biological specimens from living and deceased persons. Methods: The information sources used to prepare this review were PubMed, Scopus, and Web-of-Science. These databases were searched using keywords c-hydroxybutyrate (GHB), blood, urine, alternative specimens, non-conventional biological matrices, saliva, oral fluid, sweat, hair, vitreous humor (VH), brain, cerebrospinal fluid (CSF), dried blood spots (DBS), breast milk, and various combinations thereof. The resulting 4228 references were screened to exclude duplicates, which left 1980 articles for further consideration. These publications were carefully evaluated by taking into account the main aims of the review and 143 scientific papers were considered relevant. Analytical methods: The analytical methods used to determine c-hydroxybutyric acid in blood and other biological specimens make use of gas-or liquid-chromatography coupled to mass spectrometry. These hyphenated techniques are accurate, precise, and specific for their intended purposes and the lower limit of quantitation in blood and other specimens is 0.5 mg/L or less. Human pharmacokinetics: GHB is rapidly absorbed from the gut and distributes into the total body water compartment. Only a small fraction of the dose (1-2%) is excreted unchanged in the urine. The plasma elimination half-life of c-hydroxybutyric acid is short, being only about 0.5-0.9 h, which requires timely sampling of blood and other biological specimens for clinical and forensic analysis. Endogenous concentrations of GHB in blood: GHB is both an endogenous metabolite and a drug of abuse, which complicates interpretation of the laboratory results of analysis. Moreover, the concentrations of GHB in blood and other specimens tend to increase after sampling, especially in autopsy cases. This requires the use of practical "cut-off" concentrations to avoid reporting false positive results. These cutoffs are different for different biological specimen types. Concentrations of GHB in clinical and forensic practice: As a recreational drug GHB is predominantly used by young males (94%) with a mean age of 27.1 years. The mean (median) and range of concentrations in blood from apprehended drivers was 90 mg/L (82 mg/L) and 8-600 mg/L, respectively. The concentration distributions in blood taken from living and deceased persons overlapped, although the mean (median) and range of concentrations were higher in intoxication deaths; 640 mg/ L (280 mg/L) and 30-9200 mg/L, respectively. Analysis of GHB in alternative specimens: All bi...

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“…Nevertheless, the stability of GHB has been only partially or not investigated during method validation in many previous studies [68,[82][83][84]. In previous studies, EnGHB was stable in blood or urine at −20 °C with fluoride preservation, while an increase in GHB was observed at a higher temperature (4 °C) and/or without preservatives [67,85]. In another study, the concentration of EnGHB increased by greater than 2fold at 4 °C for 7 or 14 days and to more than 110% at −20 °C for 7 or 14 days.…”

Section: Analytical Issues Of Ghb In Biological Samplesmentioning

confidence: 99%

Metabolic Alterations Associated with γ-Hydroxybutyric Acid and the Potential of Metabolites as Biomarkers of Its Exposure

et al. 2021

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γ-Hydroxybutyric acid (GHB) is an endogenous short chain fatty acid that acts as a neurotransmitter and neuromodulator in the mammalian brain. It has often been illegally abused or misused due to its strong anesthetic effect, particularly in drug-facilitated crimes worldwide. However, proving its ingestion is not straightforward because of the difficulty in distinguishing between endogenous and exogenous GHB, as well as its rapid metabolism. Metabolomics and metabolism studies have recently been used to identify potential biomarkers of GHB exposure. This mini-review provides an overview of GHB-associated metabolic alterations and explores the potential of metabolites for application as biomarkers of GHB exposure. For this, we discuss the biosynthesis and metabolism of GHB, analytical issues of GHB in biological samples, alterations in metabolic pathways, and changes in the levels of GHB conjugates in biological samples from animal and human studies. Metabolic alterations in organic acids, amino acids, and polyamines in urine enable discrimination between GHB-ingested animals or humans and controls. The potential of GHB conjugates has been investigated in a variety of clinical settings. Despite the recent growth in the application of metabolomics and metabolism studies associated with GHB exposure, it remains challenging to distinguish between endogenous and exogenous GHB. This review highlights the significance of further metabolomics and metabolism studies for the discovery of practical peripheral biomarkers of GHB exposure.

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Long-term stability of GHB in post-mortem samples and samples from living persons, stored at −20°C, using fluoride preservatives

Cited by 19 publications

References 23 publications

Phase I metabolites (organic acids) of gamma‐hydroxybutyric acid–validated quantification using GC–MS and description of endogenous concentration ranges

Phase I metabolites (organic acids) of gamma‐hydroxybutyric acid–validated quantification using GC–MS and description of endogenous concentration ranges

Interpreting γ-hydroxybutyrate concentrations for clinical and forensic purposes

Metabolic Alterations Associated with γ-Hydroxybutyric Acid and the Potential of Metabolites as Biomarkers of Its Exposure

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