一种新型抗艾滋病药物2'-β-氟-2',3'-二脱氧腺苷反相高效液相色谱分析的荧光衍生化方法评估。

Evaluation of a fluorogenic derivatization method for the reversed-phase HPLC analysis of 2'-beta-fluoro-2',3'-dideoxyadenosine, a new anti-AIDS drug.

作者信息

Zhang H, Ford H, Roth J S, Kelley J A

机构信息

Laboratory of Medicinal Chemistry, Division of Basic Sciences, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, MD 21702-1201, USA.

出版信息

J Pharm Biomed Anal. 2001 May;25(2):285-97. doi: 10.1016/s0731-7085(00)00496-9.

DOI:10.1016/s0731-7085(00)00496-9

PMID:11275436

Abstract

High sensitivity (10(-7) to 10(-9) M) reversed-phase high-performance liquid chromatography (HPLC) analysis of adenine nucleosides and nucleotides, especially in a biological matrix, is difficult using only ultraviolet detection. Derivatization coupled with fluorescence detection has been investigated as a means of enhancing sensitivity for the reversed-phase HPLC analysis of 2'-beta-fluoro-2',3'-dideoxyadenosine (F-ddA), an experimental, acid-stable, anti-AIDS drug. The reaction of chloroacetaldehyde with the adenine base has been employed to form fluorescent 1,N(6)-etheno derivatives of F-ddA and 5'-deoxyadenosine, which is used as an internal standard. These derivatives give an analytically useful fluorescence emission at 416 nm after excitation at 230, 265, or 275 nm. Derivatization, fluorescence detection and reversed-phase chromatography have been optimized for the analysis of nanomolar concentrations of F-ddA in human plasma. This method has potential for the measurement of F-ddA at low concentration and in limited volume samples from in vivo biological studies.

摘要

仅使用紫外检测对腺嘌呤核苷和核苷酸进行高灵敏度（10⁻⁷至10⁻⁹ M）的反相高效液相色谱（HPLC）分析很困难，尤其是在生物基质中。作为提高2'-β-氟-2'，3'-二脱氧腺苷（F-ddA，一种实验性的、酸稳定的抗艾滋病药物）反相HPLC分析灵敏度的一种方法，已经研究了衍生化结合荧光检测。氯乙醛与腺嘌呤碱的反应已被用于形成F-ddA和用作内标的5'-脱氧腺苷的荧光1，N⁶-乙烯基衍生物。这些衍生物在230、265或275 nm激发后，在416 nm处产生分析上有用的荧光发射。衍生化、荧光检测和反相色谱已针对人血浆中纳摩尔浓度的F-ddA分析进行了优化。该方法具有在体内生物学研究的低浓度和有限体积样品中测量F-ddA的潜力。