Use of SPE and LC/TIS/MS/MS for rapid detection and quantitation of ketamine and its metabolite, norketamine, in urine

Wang, Kaung-Chaun; Shih, Tuo‐Shi; Cheng, Sheaw-Guey

doi:10.1016/j.forsciint.2004.03.031

Cited by 55 publications

(35 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Most assays for ketamine and metabolites in biosamples are based on chromatographic techniques [9][10][11][12], including those for enantioselective resolution [13][14][15][16][17][18], and are targeted to ketamine, norketamine and in certain cases also 5,6-dehydronorketamine. Only a few studies are available reporting the determination of hydroxylated ketamine and norketamine metabolites in biosamples.…”

Section: Introductionmentioning

confidence: 99%

CE provides evidence of the stereoselective hydroxylation of norketamine in equines

et al. 2009

View full text Add to dashboard Cite

CE provides evidence of the stereoselective hydroxylation of norketamine in equinesCE with multiple isomer sulfated-CD as selector was used for the simultaneous analysis of the stereoisomers of ketamine, norketamine, 5,6-dehydronorketamine and hydroxylated metabolites of norketamine in liquid/liquid extracts of (i) in vitro incubations with ketamine or norketamine and equine liver microsomes and (ii) plasma and urine of ponies receiving a target-controlled infusion of ketamine under isoflurane anesthesia. Hydroxynorketamine metabolites with the hydroxy group at the cyclohexanone ring could be shown to be formed stereoselectively both in vitro and in vivo. Due to the lack of standard compounds, urinary extracts were fractionated by HPLC followed by characterization of the collected fractions with CE and LC-MS n with 0.7 mmu mass discrimination. Comparison of LC-MS n data obtained with the fractions, an in vitro microsomal sample, and both pony urine and hydrolyzed pony urine led to the identification of four hydroxylated norketamine metabolites with hydroxylation at the cyclohexanone ring, two with hydroxylation at the aromatic ring and four hydroxylated metabolites of ketamine. Due to the lower detection sensitivity, only the four hydroxynorketamine metabolites with hydroxylation at the cyclohexanone ring were observed by CE. The data suggest that demethylation of ketamine followed by hydroxylation of norketamine at the cyclohexanone ring is the major metabolic pathway in equine species and that the ketamine metabolism is highly stereoselective.

show abstract

Section: Introductionmentioning

confidence: 99%

CE provides evidence of the stereoselective hydroxylation of norketamine in equines

et al. 2009

View full text Add to dashboard Cite

show abstract

“…This has led to several groups reporting the use [9,10] and need [11,12] for highly sensitive assays that can provide sufficient retrospective detection for ketamine together with other classes of fast acting sedative drugs. Many procedures have been described for detecting ketamine and its active metabolite norketamine in human urine, with varying limits of detection, ranging from 0.5 ng/mL to 25 ng/mL for both compounds [13][14][15][16][17][18][19] but only one group describe a method with an LOD of 0.05 ng/mL for norketamine, which was used to assay samples following intravenous infusion of ketamine for therapeutic purposes [13]. Despite these investigations on the analysis of ketamine, there is a paucity of data regarding the urinary elimination of ketamine and norketamine following oral administration.…”

Section: Introductionmentioning

confidence: 99%

Detection of ketamine and its metabolites in urine by ultra high pressure liquid chromatography–tandem mass spectrometry

Parkin

Turfus

Smith

et al. 2008

Journal of Chromatography B

View full text Add to dashboard Cite

“…It is also used recreationally as a "club drug" (Degenhardt et al, 2005;Kenyon et al, 2005;Leong et al, 2005), and there is a concern that ketamine may be used to facilitate sexual assault. Methods developed to analyze ketamine and norketamine in various biological matrices are widespread (Bolze and Boulieu, 1998;Yanagihara et al, 2003;Lua and Lin, 2004;Adamowicz and Kala, 2005;Leong et al, 2005;Negrusz et al, 2005;Wang et al, 2005;Apollonio et al, 2006;Xiang et al, 2006) but the analysis of the secondary metabolite dehydronorketamine, and hydroxylated metabolite (both intact and conjugated), is much less common (Adams et al, 1981;Bolze and Boulieu, 1998;Hijazi et al, 2001). At the time of this work, the lack of commercial standards has hindered analysis of these metabolites, whose identification requires custom synthesis (Huang et al, 2005;Wu et al, 2007).…”

mentioning

confidence: 99%

Use of Human Microsomes and Deuterated Substrates: An Alternative Approach for the Identification of Novel Metabolites of Ketamine by Mass Spectrometry

et al. 2009

View full text Add to dashboard Cite

ABSTRACT:In vitro biosynthesis using pooled human liver microsomes was applied to help identify in vivo metabolites of ketamine by liquid chromatography (LC)-tandem mass spectrometry. Microsomal synthesis produced dehydronorketamine, seven structural isomers of hydroxynorketamine, and at least five structural isomers of hydroxyketamine. To aid identification, stable isotopes of the metabolites were also produced from tetra-deuterated isotopes of ketamine or norketamine as substrates. Five metabolites (three hydroxynorketamine and two hydroxyketamine isomers) gave chromatographically resolved components with product ion spectra indicating the presence of a phenolic group, with phenolic metabolites being further substantiated by selective liquid-liquid extraction after adjustments to the pH. Two glucuronide conjugates of hydroxynorketamine were also identified. Analysis by LCcoupled ion cyclotron resonance mass spectrometry gave unique masses in accordance with the predicted elemental composition. The metabolites, including the phenols, were subsequently confirmed to be present in urine of subjects after oral ketamine administration, as facilitated by the addition of deuterated metabolites generated from the in vitro biosynthesis. To our knowledge, phenolic metabolites of ketamine, including an intact glucuronide conjugate, are here reported for the first time. The use of biologically synthesized deuterated material as an internal chromatographic and mass spectrometric marker is a viable approach to aid in the identification of metabolites. Metabolites that have particular diagnostic value can be selected as candidates for chemical synthesis of standards.

show abstract

Use of SPE and LC/TIS/MS/MS for rapid detection and quantitation of ketamine and its metabolite, norketamine, in urine

Cited by 55 publications

References 16 publications

CE provides evidence of the stereoselective hydroxylation of norketamine in equines

CE provides evidence of the stereoselective hydroxylation of norketamine in equines

Detection of ketamine and its metabolites in urine by ultra high pressure liquid chromatography–tandem mass spectrometry

Use of Human Microsomes and Deuterated Substrates: An Alternative Approach for the Identification of Novel Metabolites of Ketamine by Mass Spectrometry

Contact Info

Product

Resources

About