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Home / Equine Research Bank / Electrophoresis / Enantioselective CE-MS analysis of ketamine metabolites in u...
Electrophoresis2022; 44(1-2); 125-134; doi: 10.1002/elps.202200175

Enantioselective CE-MS analysis of ketamine metabolites in urine.

Abstract: The chiral drug ketamine has long-lasting antidepressant effects with a fast onset and is also suitable to treat patients with therapy-resistant depression. The metabolite hydroxynorketamine (HNK) plays an important role in the antidepressant mechanism of action. Hydroxylation at the cyclohexanone ring occurs at positions 4, 5, and 6 and produces a total of 12 stereoisomers. Among those, the four 6HNK stereoisomers have the strongest antidepressant effects. Capillary electrophoresis with highly sulfated γ-cyclodextrin (CD) as a chiral selector in combination with mass spectrometry (MS) was used to develop a method for the enantioselective analysis of HNK stereoisomers with a special focus on the 6HNK stereoisomers. The partial filling approach was applied in order to avoid contamination of the MS with the chiral selector. Concentration of the chiral selector and the length of the separation zone were optimized. With 5% highly sulfated γ-CD in 20 mM ammonium formate with 10% formic acid and a 75% filling the four 6HNK stereoisomers could be separated with a resolution between 0.79 and 3.17. The method was applied to analyze fractionated equine urine collected after a ketamine infusion and to screen the fractions as well as unfractionated urine for the parent drug ketamine and other metabolites, including norketamine and dehydronorketamine.
© 2022 The Authors. Electrophoresis published by Wiley-VCH GmbH.
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Publication Date: 2022-12-09 PubMed ID: 36398998PubMed Central: PMC10108174DOI: 10.1002/elps.202200175Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers of this paper developed a method to analyze the various forms of a ketamine metabolite, hydroxynorketamine (HNK), using capillary electrophoresis with highly sulfated γ-cyclodextrin as a chiral selector in combination with mass spectrometry (MS). They specifically focused on the 6HNK stereoisomers, which are believed to have the strongest antidepressant effects among ketamine’s 12 stereoisomers.

Hydroxynorketamine and its importance in the antidepressant effects of ketamine

  • Ketamine, a chiral drug, is known for its rapid and lasting antidepressant effects and is helpful in treating patients who are resistant to other forms of antidepressant therapy.
  • It is metabolized in the body to produce a compound known as hydroxynorketamine (HNK).
  • HNK plays a significant role in ketamine’s antidepressant effects, and it is formed through a hydroxylation process which produces a total of 12 stereoisomers (3D arrangements of molecules and atoms).
  • Of these 12 HNK stereoisomers, the ones that exhibited the strongest antidepressant effects were the four 6HNK stereoisomers. Hence, the focus on this study is on these four 6HNK stereoisomers.

Development of a method to analyze HNK stereoisomers via capillary electrophoresis and mass spectrometry

  • The researchers used a scientific technique called capillary electrophoresis assisted by a chiral selector (γ-cyclodextrin) to separate and analyze the various HNK stereoisomers. This was complemented by mass spectrometry to identify and quantify these stereoisomers.
  • Importantly, they applied a ‘partial filling approach’ to prevent contamination of the mass spectrometer detector with the γ-cyclodextrin chiral selector. This approach allows for the sample and selector to be introduced separately into the capillary and to remain separate during the procedure.
  • Various parameters were optimized during the process, including the concentration of the γ-cyclodextrin and the length of the separation zone or capillary.

Results and Applications

  • The developed method allowed for the separation of the four 6HNK stereoisomers with a resolution ranging from 0.79 to 3.17.
  • The researchers were able to apply the method to analyze fractionated equine urine collected after a ketamine infusion.
  • They also used it to identify not just the 6HNK stereoisomers, but other metabolites as well. This included the parent drug ketamine itself, as well as norketamine and dehydronorketamine.

Cite This Article

APA
Sandbaumhüter FA, Aerts JT, Theurillat R, Andrén PE, Thormann W, Jansson ET. (2022). Enantioselective CE-MS analysis of ketamine metabolites in urine. Electrophoresis, 44(1-2), 125-134. https://doi.org/10.1002/elps.202200175

Publication

Electrophoresis
ISSN: 1522-2683
NlmUniqueID: 8204476
Country: Germany
Language: English
Volume: 44
Issue: 1-2
Pages: 125-134

Researcher Affiliations

Sandbaumhüter, Friederike A
  • Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
Aerts, Jordan T
  • Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
Theurillat, Regula
  • Clinical Pharmacology Laboratory, Institute for Infectious Diseases, University of Bern, Bern, Switzerland.
Andrén, Per E
  • Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
  • Science for Life Laboratory, Spatial Mass Spectrometry, Uppsala University, Uppsala, Sweden.
Thormann, Wolfgang
  • Clinical Pharmacology Laboratory, Institute for Infectious Diseases, University of Bern, Bern, Switzerland.
Jansson, Erik T
  • Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.

MeSH Terms

  • Animals
  • Horses
  • Ketamine
  • Stereoisomerism
  • Mass Spectrometry
  • Electrophoresis, Capillary / methods
  • Sulfates

Conflict of Interest Statement

The authors have declared no conflict of interest.

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