Development of HPLC-MS/MS Method for Simultaneous Detection of Esketamine and Norketamine: Application to Pharmacokinetics Drug Interactions Affected by Dexmedetomidine.

IF 5.1 2区医学 Q1 CHEMISTRY, MEDICINAL

Drug Design, Development and Therapy Pub Date : 2025-09-22 eCollection Date: 2025-01-01 DOI:10.2147/DDDT.S553389

Wei Zhou, Zhe Guo, Xianghan Zhang, Wenjiong Wang, Zhongfeng Zheng, Xaingjun Qiu

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引用次数: 0

Abstract

Purpose: Dexmedetomidine (DEX) and esketamine (ESK) are often used together during anesthesia. This study aimed to establish a sensitive and reliable HPLC-MS/MS method for simultaneous quantification of ESK and its active metabolite norketamine (NORK) in beagle dog plasma and to investigate the pharmacokinetic drug-drug interactions (DDIs) between DEX and ESK/NORK.

Methods: A simple protein precipitation method using acetonitrile was applied for plasma sample preparation. After chromatographic separation, analytes were detected by HPLC-MS/MS in positive ion mode using multiple reaction monitoring (MRM). The mass transitions were m/z 238.10→125.10 for ESK, m/z 224.10→125.10 for NORK, and m/z 354.20→209.00 for the internal standard (proadifen). Six beagle dogs were intramuscularly administered 1 mg/kg ESK alone in the first period (ESK group). After a washout, the same dogs received intravenous DEX (2 µg/kg) for 7 consecutive days, followed by co-administration of ESK (DEX+ESK group). The pharmacokinetic parameters of ESK and NORK were calculated using DAS software. Independent-sample t test was used to compare the differences of pharmacokinetic parameters between ESK group and DEX+ESK group, and P < 0.05 indicated a statistically significant difference.

Results: Both ESK and NORK exhibited good linearity within the concentration range of 1-400 ng/mL, and the methodological validation met the requirements. When ESK was used in combination with DEX, the main pharmacokinetic parameters of ESK and NORK changed, the C_max, AUC_(0-24) and AUC_(0-∞) of ESK increased, the C_max, of NORK decreased, and the AUC_(0-12) and AUC_(0-∞) of NORK increased too.

Conclusion: A novel HPLC-MS/MS method was developed and validated and successfully applied to simultaneously quantify ESK and NORK in beagle dog plasma. The pharmacokinetic DDI results indicate that DEX could inhibit the metabolism of ESK, alter pharmacokinetic characteristics of ESK and its metabolite NORK, and significantly increase the systemic exposure of both ESK and NORK.

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HPLC-MS/MS同时检测艾氯胺酮和诺氯胺酮的方法建立：右美托咪定影响药物相互作用的药代动力学研究

目的：右美托咪定（DEX）和艾氯胺酮（ESK）常用于麻醉。本研究旨在建立一种灵敏可靠的HPLC-MS/MS同时定量比格犬血浆中ESK及其活性代谢物诺氯胺酮（NORK）的方法，并探讨DEX与ESK/NORK之间的药动学相互作用（ddi）。方法：采用简单的乙腈蛋白沉淀法制备血浆样品。色谱分离后，采用多反应监测（MRM），在正离子模式下进行HPLC-MS/MS检测。ESK的质量转变为m/z 238.10→125.10，NORK的质量转变为m/z 224.10→125.10，内标（proadifen）的质量转变为m/z 354.20→209.00。6只beagle犬在第一期（ESK组）单独肌肉注射1 mg/kg ESK。洗脱期结束后，连续7天静脉注射DEX(2µg/kg)，然后联合给药ESK （DEX+ESK组）。采用DAS软件计算ESK和NORK的药动学参数。采用独立样本t检验比较ESK组与DEX+ESK组药代动力学参数的差异，P < 0.05为差异有统计学意义。结果：ESK和NORK在1 ~ 400 ng/mL浓度范围内线性良好，方法学验证符合要求。ESK与DEX联用时，ESK和NORK的主要药动学参数发生变化，ESK的Cmax、AUC（0-24）和AUC（0-∞）升高，NORK的Cmax降低，NORK的AUC（0-12）和AUC（0-∞）也升高。结论：建立了一种高效液相色谱-质谱联用（HPLC-MS/MS）定量比格犬血浆中ESK和NORK的方法。药代动力学DDI结果表明，DEX可抑制ESK的代谢，改变ESK及其代谢物NORK的药代动力学特征，并显著增加ESK和NORK的全身暴露量。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Drug Design, Development and Therapy CHEMISTRY, MEDICINAL-PHARMACOLOGY & PHARMACY

CiteScore

9.00

自引率

0.00%

发文量

382

审稿时长

>12 weeks

期刊介绍： Drug Design, Development and Therapy is an international, peer-reviewed, open access journal that spans the spectrum of drug design, discovery and development through to clinical applications. The journal is characterized by the rapid reporting of high-quality original research, reviews, expert opinions, commentary and clinical studies in all therapeutic areas. Specific topics covered by the journal include: Drug target identification and validation Phenotypic screening and target deconvolution Biochemical analyses of drug targets and their pathways New methods or relevant applications in molecular/drug design and computer-aided drug discovery* Design, synthesis, and biological evaluation of novel biologically active compounds (including diagnostics or chemical probes) Structural or molecular biological studies elucidating molecular recognition processes Fragment-based drug discovery Pharmaceutical/red biotechnology Isolation, structural characterization, (bio)synthesis, bioengineering and pharmacological evaluation of natural products** Distribution, pharmacokinetics and metabolic transformations of drugs or biologically active compounds in drug development Drug delivery and formulation (design and characterization of dosage forms, release mechanisms and in vivo testing) Preclinical development studies Translational animal models Mechanisms of action and signalling pathways Toxicology Gene therapy, cell therapy and immunotherapy Personalized medicine and pharmacogenomics Clinical drug evaluation Patient safety and sustained use of medicines.