Postmortem distribution of ropivacaine and its metabolite in human body fluids and solid tissues by GC–MS/MS using standard addition method

IF 2.8 4区医学 Q2 TOXICOLOGY Forensic Toxicology Pub Date : 2024-09-16 DOI:10.1007/s11419-024-00695-z

Xiaolong Zhang, Shuyun Wang, Yuxuan Chen, Jie Gu, Mengchao Wang, Yinyin Dai, Kundi Zhao, Yue Wang, Amin Wurita, Koutaro Hasegawa

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Abstract

Purpose

An analytical method was developed for determining ropivacaine and its main metabolite, 3-hydroxyropivacaine in biomedical samples using gas chromatography-tandem mass spectrometry (GC–MS/MS). Then, this established method was applied to investigate the distribution of ropivacaine and its metabolite in human fluids and solid tissues obtained from an authentic case ropivacaine involved.

Methods

The fluid sample was added acetonitrile, and solid tissue was homogenized using a freezer mill and then added into acetonitrile. Then, an internal standard solution was added to the mixtures. The mixture was centrifuged at 12,000 × g for 5 min, and the upper layer of acetonitrile was transferred to magnesium sulfate and octadecyl silica (C18) gel for cleaning up the sample. After centrifugation, the upper layer was then evaporated to dryness with nitrogen, and dissolved with methanol, then injected into the GC–MS/MS system.

Results

The coefficients of determination (r²) of constructed calibration curves were all greater than 0.999. The limits of detection for ropivacaine and 3-hydroxyropivacaine in target samples were 15 ng/mL and 10 ng/mL, respectively. The recovery rates of ropivacaine and 3-hydroxyropivacaine ranged from 97.6% to 103% and from 96.5% to 104%, respectively. The inter-day precision values of ropivacaine and 3-hydroxyropivacaine were not greater than 6.25% and 7.98%, respectively, and the inter-day trueness values were not greater than 6.90% and 8.33%, respectively; the intra-day precision and trueness values of ropivacaine and 3-hydroxyropivacaine were not greater than 3.20%, 6.78%, 7.84% and 8.99%, respectively.

Conclusions

GC–MS/MS method for simultaneous detection and quantification of ropivacaine and 3-hydroxyropivacaine in biological samples was successfully developed. The method could also be applied to samples obtained from an authentic case; their distribution among tested fluids and solid tissues were also measured. This is the first report on the distribution of ropivacaine and its major metabolite 3-hydroxyropivacaine in a human case.

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采用标准添加法，通过气相色谱-质谱/质谱分析罗哌卡因及其代谢物在人体液和固体组织中的尸后分布情况

目的建立了一种气相色谱-串联质谱（GC-MS/MS）测定生物医学样品中罗哌卡因及其主要代谢物3-羟基丙哌卡因的分析方法。方法液体样品加入乙腈，固体组织用冷冻粉碎机匀浆后加入乙腈。然后在混合物中加入内标溶液。将混合物在 12,000 × g 转速下离心 5 分钟，然后将上层乙腈转移到硫酸镁和十八烷基硅胶（C18）凝胶上进行样品净化。结果构建的校正曲线的测定系数（r2）均大于 0.999。目标样品中罗哌卡因和 3-hydroxyropivacaine 的检出限分别为 15 ng/mL 和 10 ng/mL。罗哌卡因和 3-羟基丙哌卡因的回收率分别为 97.6% 至 103% 和 96.5% 至 104%。罗哌卡因和 3-羟基丙哌卡因的日间精密度值分别不大于 6.25% 和 7.98%，日间真实度值分别不大于 6.90% 和 8.33%；罗哌卡因和 3-羟基丙哌卡因的日内精密度值和真实度值分别不大于 3.结论成功建立了同时检测和定量生物样品中罗哌卡因和3-羟基丙哌卡因的GC-MS/MS方法。该方法还可应用于从真实病例中获得的样本，并测定了它们在被测体液和固体组织中的分布。这是首次报道罗哌卡因及其主要代谢物 3-羟基丙哌卡因在人体病例中的分布情况。

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来源期刊

Forensic Toxicology TOXICOLOGY-

CiteScore

5.80

自引率

9.10%

发文量

审稿时长

3 months

期刊介绍： The journal Forensic Toxicology provides an international forum for publication of studies on toxic substances, drugs of abuse, doping agents, chemical warfare agents, and their metabolisms and analyses, which are related to laws and ethics. It includes original articles, reviews, mini-reviews, short communications, and case reports. Although a major focus of the journal is on the development or improvement of analytical methods for the above-mentioned chemicals in human matrices, appropriate studies with animal experiments are also published. Forensic Toxicology is the official publication of the Japanese Association of Forensic Toxicology (JAFT) and is the continuation of the Japanese Journal of Forensic Toxicology (ISSN 0915-9606).