体积吸收微取样首次与电膜萃取相结合：全血样本中多柔比星及其代谢物的案例分析

IF 5.7 2区化学 Q1 CHEMISTRY, ANALYTICAL

Analytica Chimica Acta Pub Date : 2024-11-22 DOI:10.1016/j.aca.2024.343459

Adam Reguli , Hana Bavlovič Piskáčková , Olga Lenčová-Popelová , Petra Kollárová-Brázdová , Martin Štěrba , Stig Pedersen-Bjergaard , Petra Štěrbová-Kovaříková

{"title":"体积吸收微取样首次与电膜萃取相结合：全血样本中多柔比星及其代谢物的案例分析","authors":"Adam Reguli , Hana Bavlovič Piskáčková , Olga Lenčová-Popelová , Petra Kollárová-Brázdová , Martin Štěrba , Stig Pedersen-Bjergaard , Petra Štěrbová-Kovaříková","doi":"10.1016/j.aca.2024.343459","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>Microsampling of biological fluids followed by innovative sample pre-treatment reflects trends in bioanalytical chemistry. Volumetric absorptive microsampling (VAMS) enables exact whole blood volume collection and reduces the impact of hematocrit on the assay. In animal studies, it complies with the 3R principles (refine, reduce, replace). It allows for a gentle bleeding technique and a reduction in the number of laboratory animals by enabling ethically acceptable repeated blood collection from a single animal. Treating VAMS tips with electromembrane extraction (EME) in 96-well format offers a smart combination of non-invasive, low-volume blood collection with effective, environmentally friendly sample clean-up.</div></div><div><h3>Results</h3><div>This study introduces the first application of EME in 96-well format for direct isolation of analytes from 10 μL of whole blood collected onto a VAMS device. Doxorubicin, a clinically used anticancer drug also utilized in cancer/cardio-oncology research involving rodents, where microsampling offers important advantages, and its metabolite doxorubicinol, were selected as relevant analytes. The optimized EME yielded reproducible recoveries for both analytes regardless of hematocrit levels, different anticoagulants, or free multivalent ions in the sample. Compared to conventional VAMS tips treatment, EME reduced matrix effects, increased throughput, and an environmental friendliness of the extraction. The EME followed by the UHPLC-MS/MS assay was validated for both analytes in whole blood absorbed onto VAMS tips. The same protocol was implemented to treat plasma to determine the blood-to-plasma ratio of the analytes in the same experiments. The practical utility was demonstrated by analyzing real samples collected from the doxorubicin-treated nude mice.</div></div><div><h3>Significance</h3><div>The study offers a novel assay combining whole blood microsampling and sample clean-up in microextraction scale for preclinical pharmacokinetic studies with doxorubicin in rodents and for pharmacokinetic/pharmacodynamic modeling. This advancement in bioanalytical chemistry promotes scalable environmentally friendly procedures compatible with the 3R ethical principles in animal studies. Moreover, the concept of direct VAMS tips treatment with EME may also be easily translatable to clinical settings.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1335 ","pages":"Article 343459"},"PeriodicalIF":5.7000,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Volumetric absorptive microsampling meets electromembrane extraction for the first time: Case example of doxorubicin and its metabolite in whole blood samples\",\"authors\":\"Adam Reguli , Hana Bavlovič Piskáčková , Olga Lenčová-Popelová , Petra Kollárová-Brázdová , Martin Štěrba , Stig Pedersen-Bjergaard , Petra Štěrbová-Kovaříková\",\"doi\":\"10.1016/j.aca.2024.343459\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><div>Microsampling of biological fluids followed by innovative sample pre-treatment reflects trends in bioanalytical chemistry. Volumetric absorptive microsampling (VAMS) enables exact whole blood volume collection and reduces the impact of hematocrit on the assay. In animal studies, it complies with the 3R principles (refine, reduce, replace). It allows for a gentle bleeding technique and a reduction in the number of laboratory animals by enabling ethically acceptable repeated blood collection from a single animal. Treating VAMS tips with electromembrane extraction (EME) in 96-well format offers a smart combination of non-invasive, low-volume blood collection with effective, environmentally friendly sample clean-up.</div></div><div><h3>Results</h3><div>This study introduces the first application of EME in 96-well format for direct isolation of analytes from 10 μL of whole blood collected onto a VAMS device. Doxorubicin, a clinically used anticancer drug also utilized in cancer/cardio-oncology research involving rodents, where microsampling offers important advantages, and its metabolite doxorubicinol, were selected as relevant analytes. The optimized EME yielded reproducible recoveries for both analytes regardless of hematocrit levels, different anticoagulants, or free multivalent ions in the sample. Compared to conventional VAMS tips treatment, EME reduced matrix effects, increased throughput, and an environmental friendliness of the extraction. The EME followed by the UHPLC-MS/MS assay was validated for both analytes in whole blood absorbed onto VAMS tips. The same protocol was implemented to treat plasma to determine the blood-to-plasma ratio of the analytes in the same experiments. The practical utility was demonstrated by analyzing real samples collected from the doxorubicin-treated nude mice.</div></div><div><h3>Significance</h3><div>The study offers a novel assay combining whole blood microsampling and sample clean-up in microextraction scale for preclinical pharmacokinetic studies with doxorubicin in rodents and for pharmacokinetic/pharmacodynamic modeling. This advancement in bioanalytical chemistry promotes scalable environmentally friendly procedures compatible with the 3R ethical principles in animal studies. Moreover, the concept of direct VAMS tips treatment with EME may also be easily translatable to clinical settings.</div></div>\",\"PeriodicalId\":240,\"journal\":{\"name\":\"Analytica Chimica Acta\",\"volume\":\"1335 \",\"pages\":\"Article 343459\"},\"PeriodicalIF\":5.7000,\"publicationDate\":\"2024-11-22\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analytica Chimica Acta\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0003267024012601\",\"RegionNum\":2,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytica Chimica Acta","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0003267024012601","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}

引用次数: 0

摘要

背景对生物液体进行微采样，然后进行创新的样品预处理，这反映了生物分析化学的发展趋势。体积吸收式微采样（VAMS）可实现精确的全血量采集，减少血细胞比容对测定的影响。在动物实验中，它符合 3R 原则（完善、减少、替代）。它允许采用温和的放血技术，并通过从单个动物重复采血来减少实验动物的数量，这在伦理上是可以接受的。用 96 孔格式的电膜萃取（EME）技术处理 VAMS 顶端，实现了无创、低容量血液采集与有效、环保的样品清理的巧妙结合。结果本研究首次将 96 孔格式的 EME 应用于从收集到 VAMS 设备上的 10 μL 全血中直接分离分析物。临床上常用的抗癌药物多柔比星及其代谢物多柔比星醇被选为相关分析物，多柔比星也被用于涉及啮齿类动物的癌症/心脑肿瘤研究中，微量取样在这方面具有重要优势。无论血细胞比容水平、不同的抗凝剂或样品中的游离多价离子如何变化，优化后的 EME 对这两种分析物的回收率都是可重复的。与传统的 VAMS 提示处理相比，EME 减少了基质效应，提高了通量，并使萃取过程更加环保。在 EME 之后进行的超高效液相色谱-质谱/多质谱检测对吸附在 VAMS吸头上的全血中的两种分析物进行了验证。在相同的实验中，采用相同的方案处理血浆，以确定血液与血浆中分析物的比率。通过分析从多柔比星处理过的裸鼠身上采集的真实样本，证明了该方法的实用性。该研究提供了一种新型测定方法，它结合了全血微量取样和微萃取规模的样本净化，可用于啮齿动物多柔比星的临床前药代动力学研究以及药代动力学/药效学建模。生物分析化学的这一进步促进了符合动物研究 3R 伦理原则的可扩展环境友好型程序。此外，用 EME 直接进行 VAMS 提示治疗的概念也很容易应用于临床。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Volumetric absorptive microsampling meets electromembrane extraction for the first time: Case example of doxorubicin and its metabolite in whole blood samples

查看原文本刊更多论文

Volumetric absorptive microsampling meets electromembrane extraction for the first time: Case example of doxorubicin and its metabolite in whole blood samples

Background

Microsampling of biological fluids followed by innovative sample pre-treatment reflects trends in bioanalytical chemistry. Volumetric absorptive microsampling (VAMS) enables exact whole blood volume collection and reduces the impact of hematocrit on the assay. In animal studies, it complies with the 3R principles (refine, reduce, replace). It allows for a gentle bleeding technique and a reduction in the number of laboratory animals by enabling ethically acceptable repeated blood collection from a single animal. Treating VAMS tips with electromembrane extraction (EME) in 96-well format offers a smart combination of non-invasive, low-volume blood collection with effective, environmentally friendly sample clean-up.

Results

This study introduces the first application of EME in 96-well format for direct isolation of analytes from 10 μL of whole blood collected onto a VAMS device. Doxorubicin, a clinically used anticancer drug also utilized in cancer/cardio-oncology research involving rodents, where microsampling offers important advantages, and its metabolite doxorubicinol, were selected as relevant analytes. The optimized EME yielded reproducible recoveries for both analytes regardless of hematocrit levels, different anticoagulants, or free multivalent ions in the sample. Compared to conventional VAMS tips treatment, EME reduced matrix effects, increased throughput, and an environmental friendliness of the extraction. The EME followed by the UHPLC-MS/MS assay was validated for both analytes in whole blood absorbed onto VAMS tips. The same protocol was implemented to treat plasma to determine the blood-to-plasma ratio of the analytes in the same experiments. The practical utility was demonstrated by analyzing real samples collected from the doxorubicin-treated nude mice.

Significance

The study offers a novel assay combining whole blood microsampling and sample clean-up in microextraction scale for preclinical pharmacokinetic studies with doxorubicin in rodents and for pharmacokinetic/pharmacodynamic modeling. This advancement in bioanalytical chemistry promotes scalable environmentally friendly procedures compatible with the 3R ethical principles in animal studies. Moreover, the concept of direct VAMS tips treatment with EME may also be easily translatable to clinical settings.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Analytica Chimica Acta 化学-分析化学

CiteScore

10.40

自引率

6.50%

发文量

1081

审稿时长

38 days

期刊介绍： Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.