Quantitative chiral and achiral determination of ketamine and its metabolites by LC–MS/MS in human serum, urine and fecal samples

Hasan, Mahmoud; Hofstetter, R.; Fassauer, Georg M.; Link, Andreas; Siegmund, Werner; Oswald, Stefan

doi:10.1016/j.jpba.2017.02.035

Cited by 46 publications

(26 citation statements)

References 31 publications

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“…14 Intriguingly, clinical studies suggest that the conversion of (R,S)-Ket may be enantioselective, as significantly higher plasma concentrations of (2R,6R)-HNK relative to (2S,6S)-HNK are observed post-ketamine administration. 42,44 One study in rats reports that sub-chronic ketamine dosing increased the clearance of (R,S)-Ket and (R,S)-NK, while producing 42 and Zarate et al 43 significant increases in the plasma concentrations of (2S,6S;2R,6R)-HNK (4.5-fold), compared to a single administration. 45 Therefore, (2S,6S;2R,6R)-HNK is a major metabolite of ketamine with a considerably longer half-life, and thus, its potential contribution to ketamine's clinical effects is a very appropriate line of research.…”

Section: Pharmacokinetic Considerationsmentioning

confidence: 99%

Hydroxynorketamine: Implications for the NMDA Receptor Hypothesis of Ketamine’s Antidepressant Action

2017

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The prevailing hypothesis of ketamine's unique antidepressant effects implicates N-methyl-d-aspartate receptor (NMDAR) inhibition-dependent enhancement of a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor-mediated transmission, activation of intracellular signalling pathways and increased synaptogenesis. Recently, however, a seminal study by Zanos et al. directly challenged the NMDAR hypothesis of ketamine with the claim that an active ketamine metabolite, (2R,6R)-hydroxynorketamine, devoid of NMDAR binding properties or key side effects of its parent compound, is both necessary and sufficient for ketamine's antidepressant effects in rodents. However, following these encouraging initial findings, one preclinical study failed to replicate the antidepressant effects of (2R,6R)-hydroxynorketamine (HNK), while others have questioned the metabolite's contribution to ketamine's therapeutic effects or argued against rejecting the NMDAR hypothesis of ketamine action. In light of these potentially paradigm-shifting, but highly controversial, findings, this review will summarise and critically evaluate the evidence for and against the NMDA receptor hypothesis of ketamine action, with a particular focus on (2R,6R)-HNK and the implications of its discovery for understanding ketamine's mechanism of action in depression. Ultimately, uncovering the molecular mechanisms underlying the therapeutic effects of ketamine and possibly (2R,6R)-HNK, will aid the development of novel and more efficacious antidepressant agents so urgently needed to address a major public health concern, and could hold potential for the treatment of other stress-related psychopathologies, including bipolar disorder, post-traumatic stress disorder and suicidality.

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Section: Pharmacokinetic Considerationsmentioning

confidence: 99%

Hydroxynorketamine: Implications for the NMDA Receptor Hypothesis of Ketamine’s Antidepressant Action

2017

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“…We previously reported on the challenges associated with the HPLC bioanalysis of ketamine metabolites, which entailed one achiral C18-phase (XTerra MS ) and two CSPs that were based on protein (Chiral-AGP for K, NK and DHNK) and polysaccharides (Lux Amylose-2 for HNK) [20]. When operated in SFC mode, the same polysaccharide-based CSP afforded more versatility, as it enantioselectively resolved all metabolites but the parent drug (K), which could not be fully separated even after optimization [19].…”

Section: Resultsmentioning

confidence: 99%

Subcritical Fluid Chromatography at Sub-Ambient Temperatures for the Chiral Resolution of Ketamine Metabolites with Rapid-Onset Antidepressant Effects

et al. 2019

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Chiral metabolites of ketamine exerting rapid-onset yet sustained antidepressant effects may be marketed directly in the future, but require chemo- and enantio-selective chromatographic methods for quality assurance and control. The chromatographic behavior of S-/R-ketamine, S-/R-norketamine, S-/R-dehydronorketamine, and (2R,6R)-/(2S,6S)-hydroxynorketamine in supercritical fluid chromatography (SFC) was investigated computationally and experimentally with the aim of identifying problematic pairs of enantiomers and parameters for chiral resolution. Retention on three different polysaccharide-based chiral stationary phases (Lux Amylose-2, i-Amylose-3, and i-Cellulose-5) provided new information on the significance of halogen atoms as halogen bond donors and hydrogen bond acceptors for enantioselectivity, which could be corroborated in silico by molecular docking studies. Modifiers inversely affected enantioselectivity and retention. Methanol yielded lower run times but superior chiral resolution compared to 2-propanol. Lower temperatures than those conventionally screened did not impair phase homogeneity but improved enantioresolution, at no cost to reproducibility. Thus, sub-ambient temperature subcritical fluid chromatography (SubFC), essentially low-temperature HPLC with subcritical CO2, was applied. The optimization of the SubFC method facilitated the chiral separation of ketamine and its metabolites, which was applied in combination with direct injection and online supercritical fluid extraction to determine the purity of pharmaceutical ketamine formulations for proof of concept.

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“…Method validation was performed in accordance to similar reported analytical approaches [13,17]. The following parameters were studied: selectivity, linearity, sensitivity, accuracy and precision, as well as recovery and matrix effects.…”

Section: Validation Of Ht-baµe-µld/lvi-gc-ms(sim) Methodologymentioning

confidence: 99%

“…In a group of presumed recreational KET users, urine concentrations of KET and NKET were 6-7744 µg L −1 and 7-7986 µg L −1 , respectively [8]. Several analytical methods have been described in the literature for the determination of KET or NKET in urine samples, mostly using high performance liquid chromatography with ultraviolet/visible detection (HPLC-UV) [9], gas chromatography (GC) coupled to either flame ionization detector (FID) [10], mass spectrometry (MS) [3,7,11], Tandem mass spectrometry (MS/MS) [12] or even liquid chromatography (LC) coupled to MS [7,8] or MS/MS [13,14]. Sample preparation techniques used in combination with these chromatographic or hyphenated systems include conventional liquid-liquid extraction (LLE) [8] or solid-phase extraction (SPE) [7,14,15], but also miniaturized techniques such as solid-phase microextraction (SPME) [16], stir bar sorptive extraction [9], hollow-fiber liquid-phase microextraction [3,10] or microextraction by packed sorbent (MEPS) [12].…”

Section: Introductionmentioning

confidence: 99%

A Fast and Validated High Throughput Bar Adsorptive Microextraction (HT-BAµE) Method for the Determination of Ketamine and Norketamine in Urine Samples

et al. 2020

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We developed, optimized and validated a fast analytical cycle using high throughput bar adsorptive microextraction and microliquid desorption (HT-BAμE-μLD) for the extraction and desorption of ketamine and norketamine in up to 100 urine samples simultaneously, resulting in an assay time of only 0.45 min/sample. The identification and quantification were carried out using large volume injection-gas chromatography-mass spectrometry operating in the selected ion monitoring mode (LVI-GC-MS(SIM)). Several parameters that could influencing HT-BAµE were assayed and optimized in order to maximize the recovery yields of ketamine and norketamine from aqueous media. These included sorbent selectivity, desorption solvent and time, as well as shaking rate, microextraction time, matrix pH, ionic strength and polarity. Under optimized experimental conditions, suitable sensitivity (1.0 μg L−1), accuracy (85.5–112.1%), precision (≤15%) and recovery yields (84.9–105.0%) were achieved. Compared to existing methods, the herein described analytical cycle is much faster, environmentally friendly and cost-effective for the quantification of ketamine and norketamine in urine samples. To our knowledge, this is the first work that employs a high throughput based microextraction approach for the simultaneous extraction and subsequent desorption of ketamine and norketamine in up to 100 urine samples simultaneously.

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Quantitative chiral and achiral determination of ketamine and its metabolites by LC–MS/MS in human serum, urine and fecal samples

Cited by 46 publications

References 31 publications

Hydroxynorketamine: Implications for the NMDA Receptor Hypothesis of Ketamine’s Antidepressant Action

Hydroxynorketamine: Implications for the NMDA Receptor Hypothesis of Ketamine’s Antidepressant Action

Subcritical Fluid Chromatography at Sub-Ambient Temperatures for the Chiral Resolution of Ketamine Metabolites with Rapid-Onset Antidepressant Effects

A Fast and Validated High Throughput Bar Adsorptive Microextraction (HT-BAµE) Method for the Determination of Ketamine and Norketamine in Urine Samples

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