Sherry Cox, Madeline Duncan, Riley Golias, Julia Cutchin, Nora Springer, Aaron Bloom, Joan Bergman
{"title":"Development and Validation of a High-Performance Liquid Chromatography Method for the Detection of Rose Bengal in Canine Plasma","authors":"Sherry Cox, Madeline Duncan, Riley Golias, Julia Cutchin, Nora Springer, Aaron Bloom, Joan Bergman","doi":"10.1007/s10337-025-04435-2","DOIUrl":null,"url":null,"abstract":"<div><p>A straightforward and precise technique for the analysis of Rose Bengal concentrations in canine plasma was established and validated using HPLC. Following a simple protein precipitation method with methanol, separation occurred on an XBridge Phenyl column with a mobile phase of 10 mM ammonium phosphate and acetonitrile (65:35, v/v). Rose Bengal and meloxicam, the internal standard, were confirmed by monitoring UV–visible wavelengths at 549 nm and 370 nm, respectively. The lower limit of quantification (LLOQ) was determined to be 25 ng mL<sup>−1</sup> in a 100 μL sample. The recovery was greater than 90% while intra-assay variability was less than 5%. Inter-assay variability was less than 7%. This method could be useful in pharmacokinetic studies.</p></div>","PeriodicalId":518,"journal":{"name":"Chromatographia","volume":"88 10","pages":"771 - 777"},"PeriodicalIF":1.3000,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chromatographia","FirstCategoryId":"92","ListUrlMain":"https://link.springer.com/article/10.1007/s10337-025-04435-2","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
A straightforward and precise technique for the analysis of Rose Bengal concentrations in canine plasma was established and validated using HPLC. Following a simple protein precipitation method with methanol, separation occurred on an XBridge Phenyl column with a mobile phase of 10 mM ammonium phosphate and acetonitrile (65:35, v/v). Rose Bengal and meloxicam, the internal standard, were confirmed by monitoring UV–visible wavelengths at 549 nm and 370 nm, respectively. The lower limit of quantification (LLOQ) was determined to be 25 ng mL−1 in a 100 μL sample. The recovery was greater than 90% while intra-assay variability was less than 5%. Inter-assay variability was less than 7%. This method could be useful in pharmacokinetic studies.
期刊介绍：
Separation sciences, in all their various forms such as chromatography, field-flow fractionation, and electrophoresis, provide some of the most powerful techniques in analytical chemistry and are applied within a number of important application areas, including archaeology, biotechnology, clinical, environmental, food, medical, petroleum, pharmaceutical, polymer and biopolymer research. Beyond serving analytical purposes, separation techniques are also used for preparative and process-scale applications. The scope and power of separation sciences is significantly extended by combination with spectroscopic detection methods (e.g., laser-based approaches, nuclear-magnetic resonance, Raman, chemiluminescence) and particularly, mass spectrometry, to create hyphenated techniques. In addition to exciting new developments in chromatography, such as ultra high-pressure systems, multidimensional separations, and high-temperature approaches, there have also been great advances in hybrid methods combining chromatography and electro-based separations, especially on the micro- and nanoscale. Integrated biological procedures (e.g., enzymatic, immunological, receptor-based assays) can also be part of the overall analytical process.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
