通过超滤测定抗生素的游离血清浓度：对实验变量的广泛评估。

IF 1.9 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2024-01-01 Epub Date: 2024-07-23 DOI:10.1080/17576180.2024.2365526

Constantin Lier, Alexander Dejaco, Alexander Kratzer, Martin G Kees, Frieder Kees, Christoph Dorn

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引用次数: 0

摘要

目的：评估实验条件对多种抗生素中通过超滤和 HPLC-DAD 分析测定的游离血清浓度的影响。材料与方法：改变相对离心力（RCF）、温度、pH 值和缓冲液，并将结果与标准方案（磷酸盐缓冲液，pH 值 7.4，37°C，1000×g）进行比较。结果：一般来说，在 10,000 × g 的条件下，未结合部分（fu）随着分子量的增加而减少，在 22°C 时更低。在非缓冲血清中，氟氯西林或丙戊酸的未结合部分（fu）增加，碱性或两性药物的未结合部分（fu）明显减少。除形成金属螯合物的药物外，使用磷酸盐或 HEPES 缓冲液也能得到相似的结果。结论保持生理 pH 值比严格保持体温更为重要。

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Free serum concentrations of antibiotics determined by ultrafiltration: extensive evaluation of experimental variables.

Aim: To assess the impact of experimental conditions on free serum concentrations as determined by ultrafiltration and HPLC-DAD analysis in a wide range of antibiotics.Materials & methods: Relative centrifugation force (RCF), temperature, pH and buffer were varied and the results compared with the standard protocol (phosphate buffer pH 7.4, 37°C, 1000 × g).Results: Generally, at 10,000 × g the unbound fraction (f_u) decreased with increasing molecular weight, and was lower at 22°C. In unbuffered serum, the f_u of flucloxacillin or valproic acid was increased, that of basic or amphoteric drugs considerably decreased. Comparable results were obtained using phosphate or HEPES buffer except for drugs which form metal chelate complexes.Conclusion: Maintaining a physiological pH is more important than strictly maintaining body temperature.

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.