高效液相色谱法测定体内与脱氧核糖核酸结合的阿糖胞苷浓度

Determination of 1-β-D-arabinofuranosylcytosine incorporation into DNA in vivo by HPLC

目的：建立一种定量测定阿糖胞苷（Ara-C）治疗期间体内的白血病细胞中与脱氧核糖核酸（DNA）结合的Ara-C含量，且无需放射性元素标记的方法。方法：从患者幼稚细胞中提取DNA，DNA的酶消化和高效液相色谱法测定Ara-C的量。结果：测定15例接受常规剂量Ara-C持续静脉榆注24h时的急性髓系白血病患儿外周血标本，与DNA结合的Ara-C的量为3．23～96．73ng·μg^-1 DNA。结论：这种方法敏感、简易、稳定可靠，可被用于测量体内与白血病细胞DNA结合的Ara-C的浓度。

OBJECTIVE To developed a quantitative procedure to determine incorporation of Ara-C into leukemic cell DNA in vivo during Ara-C therapy. METHODS The study included DNA isolation from patient myeloblasts, enzymatic digestion of the DNA,and determination of Ara-C in the digestion solution by HPLC. RESULTS Myeloblasts from 15 AML children receiving conventional-dose continuous-infusion Ara-C therapy incorporated 3.23 - 96. 73 ng·μg^-1 DNA during 24 h of therapy. CONCLUSION These findings suggest that this method is sensitive,simple and reliable,and it can be used to monitor the in vivo incorporation of Ara-C into leukemic cell DNA.