Hegedus L, Ford H, Hartman N R, Kelley J A
Laboratory of Medicinal Chemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4255, USA.
J Chromatogr B Biomed Sci Appl. 1997 Apr 25;692(1):169-79. doi: 10.1016/s0378-4347(96)00464-1.
Cyclopentenyl cytosine (CPE-C) is a synthetic carbocyclic nucleoside that possesses diverse antitumor and antiviral activity. CPE-C has been studied extensively at the preclinical level and has been evaluated in a Phase I clinical trial involving patients with solid tumors. A narrow-bore, reversed-phase HPLC method that has been developed for the sensitive measurement of CPE-C in plasma and urine in order to carry out these studies is described. Covalent solid-phase extraction based on an immobilized phenylboronic acid ligand is employed to isolate both CPE-C and endogenous ribonucleosides from the biological matrix selectively and efficiently. This is followed by isocratic elution of the extract with pH 5.0, 0.1 M ammonium formate buffer at 0.150 ml/min on a tandem, switchable, C18 narrow-bore (2.1 mm I.D.) column system in which the precolumn is automatically backflushed to eliminate late-eluting components. UV detection at 278 nm provides a limit of quantitation of 0.1 microM for CPE-C in rat and human plasma with a precision better than 4% for the range 1-20 microM in rat plasma. Application of this assay to the determination of the bolus dose plasma kinetics and disposition of 2 mg/kg CPE-C in rats is illustrated. This method is amenable to partial automation and is well-suited for the analysis of clinical samples.
环戊烯基胞嘧啶（CPE-C）是一种合成的碳环核苷，具有多种抗肿瘤和抗病毒活性。CPE-C已在临床前水平进行了广泛研究，并已在一项涉及实体瘤患者的I期临床试验中进行了评估。本文描述了一种为灵敏测定血浆和尿液中的CPE-C而开发的窄孔反相高效液相色谱法，以便开展这些研究。基于固定化苯基硼酸配体的共价固相萃取用于从生物基质中选择性、高效地分离CPE-C和内源性核糖核苷。然后，在串联、可切换的C18窄孔（内径2.1 mm）柱系统上，用pH 5.0、0.1 M甲酸铵缓冲液以0.150 ml/min的流速对提取物进行等度洗脱，在此系统中，预柱会自动反冲以消除晚洗脱成分。在278 nm处进行紫外检测，大鼠和人血浆中CPE-C的定量限为0.1 μM，在大鼠血浆中1-20 μM范围内精密度优于4%。本文举例说明了该分析方法在测定大鼠静脉推注剂量2 mg/kg CPE-C后的血浆动力学和处置情况中的应用。该方法适合部分自动化，非常适合临床样本分析。
