摘要
目的:建立一种快速、灵敏、高效的方法,检测儿童急性淋巴细胞白血病红细胞中硫鸟嘌呤核苷酸浓度。方法:应用反相高效液相色谱(RT-HPLC)技术,测定急性淋巴细胞白血病患儿红细胞内6-巯嘌呤(6-MP)及其代谢产物[硫鸟嘌呤核苷酸、6-硫代黄嘌呤(6-TX)、6-甲基巯基嘌呤(6-MMP)]浓度。色谱柱为Resolve C18,柱温40℃。流动相A为0.2%的乙酸溶液,流动相B为甲醇。梯度洗脱程序为在16min内流动相B的浓度从0上升到80%。流速1.0mL/min。检测波长为0~12.5min用290nm检测6-硫鸟嘌呤(6-TG)、6-MP和6-TX,12.5min时切换为340nm检测6-MMP。结果:进样体积为10μL时,6-TG和6-TX的线性范围为0~20nmol/mL(r=0.9993,r=0.9980)。硫鸟嘌呤核苷酸浓度与测定后第9天的白细胞计数呈负相关(P<0.01)。结论:RT-HPLC法简便、准确检测6-MP代谢物浓度,有助于个体化的治疗。
Objective To establish a HPLC method to measure thioguanine nucleotides(TGNs) concentration in children with acute lymphoblastie leukemia(ALL). Method The concentration of 6-MP(6-mereaptopurine), 6-TGNs and 6-TX in RBC were determined using reversed-phase high performance liquid chromatography (RP-HPLC) method with resolve C18 columns (250 mm× 4.6 mm,5μm) at 40℃. The mobile phase A was 0.2% ethanoie acid and mobile B was methanol. The gradient elution procedure was as follows : the mobile B concentration from 0 to 80% in 16 min with a flow rate of 1.0 mL/min.The detection wave-length was set at 290 nm from 0 to 12.5 rain to detect 6-TG, 6-MP and 6-TX , and 340 nm at 12.5 rain to detect 6-MMP. Result The good linearity was shown on 6-TG and 6-TX concentration range of 0 - 20 nmol/mL(r = 0.999 3, r = 0.998 0). In children with ALL, the TGNs level did show a negative correlation with the numbers of WBC 16 days after TGNs assay (P 〈 0.01 ). Conclusion This method is simple, rapid and suitable for clinical drug monitoring assay in individualizing 6-MP chemotherapy for children with ALL.
出处
《实用医学杂志》
CAS
北大核心
2009年第21期3688-3690,共3页
The Journal of Practical Medicine
基金
广东省医学科学技术研究基金(编号:2005288)
广州医学院博士启动基金(编号:0706067)