Zhang Xiaolong, Wang Shuyun, Chen Yuxuan, Gu Jie, Wang Mengchao, Dai Yinyin, Zhao Kundi, Wang Yue, Wurita Amin, Hasegawa Koutaro
Department of Legal Medicine, College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, China.
Durg Control Detachment Hohhot Publ Secur Bur, Hohhot, 010050, People's Republic of China.
Forensic Toxicol. 2025 Jan;43(1):163-171. doi: 10.1007/s11419-024-00695-z. Epub 2024 Sep 16.
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.
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.
The coefficients of determination (r) 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.
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.
建立一种采用气相色谱 - 串联质谱法（GC-MS/MS）测定生物医学样本中罗哌卡因及其主要代谢产物3-羟基罗哌卡因的分析方法。然后，将该方法应用于研究罗哌卡因及其代谢产物在涉及罗哌卡因真实案例的人体体液和实体组织中的分布情况。
向液体样本中加入乙腈，使用冷冻研磨机将实体组织匀浆后加入乙腈。然后，向混合物中加入内标溶液。混合物在12,000×g下离心5分钟，将上层乙腈转移至硫酸镁和十八烷基硅烷（C18）凝胶中进行样品净化。离心后，上层溶液用氮气蒸发至干，用甲醇溶解，然后注入GC-MS/MS系统。
构建的校准曲线的测定系数（r）均大于0.999。目标样本中罗哌卡因和3-羟基罗哌卡因的检测限分别为15 ng/mL和10 ng/mL。罗哌卡因和3-羟基罗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-羟基罗哌卡因的日间精密度值分别不大于6.25%和7.98%，日间真实性值分别不大于6.90%和8.33%；罗哌卡因和3-羟基罗哌卡因的日内精密度和真实性值分别不大于3.20%、6.78%、7.84%和8.99%。
成功建立了GC-MS/MS法同时检测和定量生物样本中罗哌卡因和3-羟基罗哌卡因的方法。该方法也可应用于真实案例获得的样本；还测定了它们在受试体液和实体组织中的分布。这是关于罗哌卡因及其主要代谢产物3-羟基罗哌卡因在人体案例中分布的首次报道。
