Sensitive UHPLC-MS/MS and enantioselective SFC-MS method for determination of phar-makokinetic profiles of methoxphenidine and its metabolite in biological matrices

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22310%2F24%3A43929554" target="_blank" >RIV/60461373:22310/24:43929554 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60461373:22330/24:43929554 RIV/60461373:22340/24:43929554

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Sensitive UHPLC-MS/MS and enantioselective SFC-MS method for determination of phar-makokinetic profiles of methoxphenidine and its metabolite in biological matrices

Popis výsledku v původním jazyce

Dissociative anaesthetics (particularly ketamine) have potential as next generation antidepressants, especially, for treatment of patients with treatment refractory depression. Increasing evidence accumulates that some metabolites of ketamine contribute considerably to this plausible effect. Moreover, the effect usually varies for the R- or S enantiomer thereby putting drug stereochemistry into the spotlight.1,2 On the other hand, ketamine is abused as new psychoactive substance (NPS). What’s worse, the number of his falsifications increased. One of them, methoxphenidine (MXP) has been embracing on the black market in 2013, and MXP intoxications have been reported with at least three deaths since then. Analytical methods for the simultaneous determination of MXP and its metabolite normethoxphenidine - norMXP are a prerequisite for future clinical investigations.MethodsSensitive UHPLC-MS/MS analyses were performed using LC system Dionex Ultimate 3000 (Thermo-Fisher) and the chromatographic separation was performed using column Poroshell 120, Phenyl Hexyl (Agilent). The analytes were detected by QTRAP 6500+ (AB Sciex) with a IonDrive Turbo V ion source operating in positive electrospray mode. Moreover, an enantioselective SFC-MS method was developed on Acquity UltraPerformance Convergence ChromatographyTM (UPC2) coupled to a single quadrupole (QDa) with electrospray ionization (Waters). A chiral polysaccharide stationary phase Alcyon SFC CSP Amylose-SA (YMC) was used. Fifty-six Wistar rats were subcutaneously administered 20 mg/kg of racemic MXP. Serum (2 mL) and tissues from eight animals were collected per each time point (0, 0.5, 1, 2, 4, 8 and 24 hours after administration).Preliminary DataTwo different chromatographic methods were evaluated in this study. All validation parameters met the guidelines from the European Medicines Agency (EMA). The UHPLC-MS/MS mobile phase consisted of formic acid, ammonium formate in water and acetonitrile in methanol. A gradient elution, flow rate of 0.4 mL/min and column temperature at 35°C were used. The SFC-MS mobile phase composed CO2, ethanol and ammonia in a gradient mode. The method was set with back pressure regulator on 2500 psi, flow rate at 1 mL/min and temperature at 35°C. Consequently, we have elaborated extraction methods for MXP and norMXP from Wistar rat serum and brain tissue. Protein precipitation was chosen as the best extraction method for processing blood serum samples. Due to the complex matrix, salting-out liquid-liquid extraction (SALLE) was chosen for preparation brain tissue. A wide concentration range was evaluated in case of serum (0.25 ng/mL to 2000 ng/mL) and brain (0.6 ng/g to 15000 ng/g).Following the administration of racemic MXP to Wistar rats, serum profile of MXP and norMXP was generated. The maximum concentration of MXP occurred with 0.5 and 1 hours after administration. The major metabolite, norMXP was observed in high concentrations at 1 hour after administration. There was no significant difference in concentration of MXP enantiomers per each time point. Brain profile of MXP will by also generated. Our preliminary data predicted the maximum concentration of MXP and norMXP at 0.5, 1 or 2 hours after administration. Collected data will be discussed and correlated with data from behavioral study of the National Institute of Mental Health.Novel AspectWe presented pharmacokinetic profiles of ketamine related MXP in rat serum and brain samples. The enantiomeric ratio was also determined.Conflict of Interest DisclosureThe authors declare no competing financial interest.

Název v anglickém jazyce

Sensitive UHPLC-MS/MS and enantioselective SFC-MS method for determination of phar-makokinetic profiles of methoxphenidine and its metabolite in biological matrices

Popis výsledku anglicky

Dissociative anaesthetics (particularly ketamine) have potential as next generation antidepressants, especially, for treatment of patients with treatment refractory depression. Increasing evidence accumulates that some metabolites of ketamine contribute considerably to this plausible effect. Moreover, the effect usually varies for the R- or S enantiomer thereby putting drug stereochemistry into the spotlight.1,2 On the other hand, ketamine is abused as new psychoactive substance (NPS). What’s worse, the number of his falsifications increased. One of them, methoxphenidine (MXP) has been embracing on the black market in 2013, and MXP intoxications have been reported with at least three deaths since then. Analytical methods for the simultaneous determination of MXP and its metabolite normethoxphenidine - norMXP are a prerequisite for future clinical investigations.MethodsSensitive UHPLC-MS/MS analyses were performed using LC system Dionex Ultimate 3000 (Thermo-Fisher) and the chromatographic separation was performed using column Poroshell 120, Phenyl Hexyl (Agilent). The analytes were detected by QTRAP 6500+ (AB Sciex) with a IonDrive Turbo V ion source operating in positive electrospray mode. Moreover, an enantioselective SFC-MS method was developed on Acquity UltraPerformance Convergence ChromatographyTM (UPC2) coupled to a single quadrupole (QDa) with electrospray ionization (Waters). A chiral polysaccharide stationary phase Alcyon SFC CSP Amylose-SA (YMC) was used. Fifty-six Wistar rats were subcutaneously administered 20 mg/kg of racemic MXP. Serum (2 mL) and tissues from eight animals were collected per each time point (0, 0.5, 1, 2, 4, 8 and 24 hours after administration).Preliminary DataTwo different chromatographic methods were evaluated in this study. All validation parameters met the guidelines from the European Medicines Agency (EMA). The UHPLC-MS/MS mobile phase consisted of formic acid, ammonium formate in water and acetonitrile in methanol. A gradient elution, flow rate of 0.4 mL/min and column temperature at 35°C were used. The SFC-MS mobile phase composed CO2, ethanol and ammonia in a gradient mode. The method was set with back pressure regulator on 2500 psi, flow rate at 1 mL/min and temperature at 35°C. Consequently, we have elaborated extraction methods for MXP and norMXP from Wistar rat serum and brain tissue. Protein precipitation was chosen as the best extraction method for processing blood serum samples. Due to the complex matrix, salting-out liquid-liquid extraction (SALLE) was chosen for preparation brain tissue. A wide concentration range was evaluated in case of serum (0.25 ng/mL to 2000 ng/mL) and brain (0.6 ng/g to 15000 ng/g).Following the administration of racemic MXP to Wistar rats, serum profile of MXP and norMXP was generated. The maximum concentration of MXP occurred with 0.5 and 1 hours after administration. The major metabolite, norMXP was observed in high concentrations at 1 hour after administration. There was no significant difference in concentration of MXP enantiomers per each time point. Brain profile of MXP will by also generated. Our preliminary data predicted the maximum concentration of MXP and norMXP at 0.5, 1 or 2 hours after administration. Collected data will be discussed and correlated with data from behavioral study of the National Institute of Mental Health.Novel AspectWe presented pharmacokinetic profiles of ketamine related MXP in rat serum and brain samples. The enantiomeric ratio was also determined.Conflict of Interest DisclosureThe authors declare no competing financial interest.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/VJ01010043" target="_blank" >VJ01010043: Pharmacrime – forenzní identifikace prohormonů a padělků léčiv</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů