当前位置: X-MOL 学术J. Chromatogr. B › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
A targeted ultra performance liquid chromatography – Tandem mass spectrometric assay for tyrosine and metabolites in urine and plasma: Application to the effects of antibiotics on mice
Journal of Chromatography B ( IF 3 ) Pub Date : 2020-12-22 , DOI: 10.1016/j.jchromb.2020.122511
Marine P.M. Letertre , Antonis Myridakis , Luke Whiley , Stéphane Camuzeaux , Matthew R. Lewis , Katie E. Chappell , Annie Thaikkatil , Marc-Emmanuel Dumas , Jeremy K. Nicholson , Jonathan R. Swann , Ian D. Wilson

Tyrosine plays a key role in mammalian biochemistry and defects in its metabolism (e.g., tyrosinemia, alkaptonuria etc.) have significant adverse consequences for those affected if left untreated. In addition, gut bacterially-derived p-cresol and its metabolites are of interest as a result of various effects on host xenobiotic metabolism. A fit-for-purpose quantitative ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay was developed to target and quantify tyrosine and eleven metabolites in urine and plasma. Dansylation, using dansyl chloride, was used to improve chromatographic and mass spectral properties for tyrosine and nine phenolic metabolites, with detection using positive electrospray ionisation (ESI). The sulfate and glucuronide conjugates of p-cresol, where the phenol group was blocked, were quantified intact, using negative ESI via polarity switching during the same run. Sample preparation for urine and plasma involved deproteinization by solvent precipitation (of acetonitrile:isopropyl alcohol (1:1 v/v)) followed by in situ dansylation in 96 well plates. To minimize sample and solvent usage, and maximize sensitivity, analysis was performed using microbore reversed-phase gradient UPLC on a C8 phase with a 7.5 min. cycle time. The coefficients of variation obtained were <15%, with lower limits of quantification ranging from 5 to 250 nM depending upon the analyte. The method was applied to plasma and urine samples obtained from mice placed on a high tyrosine diet with one subgroup of animals subsequently receiving antibiotics to suppress the gut microbiota. Whilst plasma profiles were largely unaffected by antibiotic treatment clear reductions in the amount of p-cresol sulfate and p-cresol glucuronide excreted in the urine were observed for these mice.



中文翻译:

靶向超高效液相色谱-串联质谱法测定尿液和血浆中的酪氨酸和代谢物:在抗生素对小鼠的影响中的应用

酪氨酸在哺乳动物的生物化学中起着关键作用,如果不加以治疗,其代谢缺陷(例如酪氨酸血症,链蛋白尿症等)会对受影响的人产生重大不利后果。另外,由于对宿主异种生物代谢的各种作用,肠内细菌衍生的甲酚及其代谢物也受到关注。开发了适用于目的的定量超高性能液相色谱串联质谱分析(UPLC-MS / MS),以靶向和定量尿液和血浆中的酪氨酸和11种代谢物。丹磺酰化,使用丹磺酰氯,用于改善酪氨酸和9种酚类代谢物的色谱和质谱性质,并使用正电喷雾电离(ESI)进行检测。硫酸盐和葡萄糖醛酸结合物p甲酚,其中苯酚基团被封,进行定量完好,使用相同的运行期间经由极性切换负ESI。尿液和血浆的样品制备涉及通过溶剂沉淀(乙腈:异丙醇(1:1 v / v))进行去蛋白,然后原位进行在96孔板中进行丹磺酰化。为了最大程度地减少样品和溶剂的使用并最大程度地提高灵敏度,使用微孔反相梯度UPLC在C8相上进行了7.5分钟的分析。周期。获得的变异系数<15%,定量下限取决于分析物,范围为5至250 nM。该方法适用于血浆和尿液样品,这些血浆和尿液样品来自高酪氨酸饮食的小鼠,其中一小组动物随后接受抗生素抑制肠道菌群。虽然血浆分布在很大程度上不受抗生素治疗的影响,但对这些小鼠观察到尿中对-甲酚硫酸盐和对-甲酚葡萄糖醛酸苷的排泄量明显减少。

更新日期:2021-01-16
down
wechat
bug