Yupeng He , Paul Miggiels , Amy Harms , Yvonne Rijksen , Renata M.C. Brandt , Wilbert P. Vermeij , Bert Wouters , Thomas Hankemeier
{"title":"人血浆和小鼠肌肉组织中酰基肉碱的全自动、高通量电萃取和分析工作流程","authors":"Yupeng He , Paul Miggiels , Amy Harms , Yvonne Rijksen , Renata M.C. Brandt , Wilbert P. Vermeij , Bert Wouters , Thomas Hankemeier","doi":"10.1016/j.aca.2025.344224","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>The labor-intensive and time-consuming nature of sample preparation poses significant challenges for bioanalysis, especially for large-scale samples characterized by limited volumes/mass, and low analyte abundance. Additionally, manual sample processing can compromise reproducibility. To overcome these limitations, automation and high-throughput methodologies are essential, highlighting the need for an automated, high-throughput sample preparation and analysis workflow.</div></div><div><h3>Results</h3><div>This study presents a fully automated, high-throughput electro-extraction (EE) platform integrated with a CTC PAL3 autosampler and liquid chromatography–mass spectrometry analyzer. The integrated platform underwent qualification, followed by optimization of EE parameters using a Design of Experiment approach. Ten acylcarnitines were selected as model analytes. The optimization models exhibited strong fits (p < 0.006, R<sup>2</sup> > 0.91). The optimized platform achieved an enrichment factor of up to 400 (an extraction recovery of up to 99 %) in designed academic samples, and was effectively implemented and evaluated using 20 μL of spiked human plasma samples. To test clinically relevant materials, the platform was utilized to study the effects of muscle tissue isolation speed on acylcarnitine stability, and to examine acylcarnitine abundance across muscle types in progeria (sarcopenia) mouse muscle. We found that the speed of muscle isolation does not affect measured levels of acylcarnitines, and detected higher acylcarnitine abundances are consistent with literature.</div></div><div><h3>Significance</h3><div>This study provides an automated, high-throughput, and cost-effective workflow enabling extraction and analysis of 120 samples per day, with a cost of <0.1 Euro per sample. It presents a significant stride towards the creation of fully-automated, high-throughput bioanalysis workflows for large-scale studies involving biomass limited samples in the foreseeable future.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1364 ","pages":"Article 344224"},"PeriodicalIF":5.7000,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A fully automated, high-throughput electro-extraction and analysis workflow for acylcarnitines in human plasma and mouse muscle tissues\",\"authors\":\"Yupeng He , Paul Miggiels , Amy Harms , Yvonne Rijksen , Renata M.C. Brandt , Wilbert P. Vermeij , Bert Wouters , Thomas Hankemeier\",\"doi\":\"10.1016/j.aca.2025.344224\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><div>The labor-intensive and time-consuming nature of sample preparation poses significant challenges for bioanalysis, especially for large-scale samples characterized by limited volumes/mass, and low analyte abundance. Additionally, manual sample processing can compromise reproducibility. To overcome these limitations, automation and high-throughput methodologies are essential, highlighting the need for an automated, high-throughput sample preparation and analysis workflow.</div></div><div><h3>Results</h3><div>This study presents a fully automated, high-throughput electro-extraction (EE) platform integrated with a CTC PAL3 autosampler and liquid chromatography–mass spectrometry analyzer. The integrated platform underwent qualification, followed by optimization of EE parameters using a Design of Experiment approach. Ten acylcarnitines were selected as model analytes. The optimization models exhibited strong fits (p < 0.006, R<sup>2</sup> > 0.91). The optimized platform achieved an enrichment factor of up to 400 (an extraction recovery of up to 99 %) in designed academic samples, and was effectively implemented and evaluated using 20 μL of spiked human plasma samples. To test clinically relevant materials, the platform was utilized to study the effects of muscle tissue isolation speed on acylcarnitine stability, and to examine acylcarnitine abundance across muscle types in progeria (sarcopenia) mouse muscle. We found that the speed of muscle isolation does not affect measured levels of acylcarnitines, and detected higher acylcarnitine abundances are consistent with literature.</div></div><div><h3>Significance</h3><div>This study provides an automated, high-throughput, and cost-effective workflow enabling extraction and analysis of 120 samples per day, with a cost of <0.1 Euro per sample. 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A fully automated, high-throughput electro-extraction and analysis workflow for acylcarnitines in human plasma and mouse muscle tissues
Background
The labor-intensive and time-consuming nature of sample preparation poses significant challenges for bioanalysis, especially for large-scale samples characterized by limited volumes/mass, and low analyte abundance. Additionally, manual sample processing can compromise reproducibility. To overcome these limitations, automation and high-throughput methodologies are essential, highlighting the need for an automated, high-throughput sample preparation and analysis workflow.
Results
This study presents a fully automated, high-throughput electro-extraction (EE) platform integrated with a CTC PAL3 autosampler and liquid chromatography–mass spectrometry analyzer. The integrated platform underwent qualification, followed by optimization of EE parameters using a Design of Experiment approach. Ten acylcarnitines were selected as model analytes. The optimization models exhibited strong fits (p < 0.006, R2 > 0.91). The optimized platform achieved an enrichment factor of up to 400 (an extraction recovery of up to 99 %) in designed academic samples, and was effectively implemented and evaluated using 20 μL of spiked human plasma samples. To test clinically relevant materials, the platform was utilized to study the effects of muscle tissue isolation speed on acylcarnitine stability, and to examine acylcarnitine abundance across muscle types in progeria (sarcopenia) mouse muscle. We found that the speed of muscle isolation does not affect measured levels of acylcarnitines, and detected higher acylcarnitine abundances are consistent with literature.
Significance
This study provides an automated, high-throughput, and cost-effective workflow enabling extraction and analysis of 120 samples per day, with a cost of <0.1 Euro per sample. It presents a significant stride towards the creation of fully-automated, high-throughput bioanalysis workflows for large-scale studies involving biomass limited samples in the foreseeable future.
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.