中国临床药理学杂志Issue(3)：257-260,4.DOI:10.13699/j.cnki.1001-6821.2016.03.020
高效液相色谱法测定人红细胞6－硫鸟嘌呤核苷酸和6－甲基巯基嘌呤核苷酸的浓度
Simultaneously determination of 6-thioguanine nucleotides and 6-methlymercaptopurine nucleotides concentrations in human red blood cells by HPLC
摘要
Abstract
Objective To establish a high performance liquid chroma-tography ( HPLC ) method for determining thiopurine active metabolites 6-thioguanine nucleotides ( 6 -TGNs ) and 6 -methlymercaptopurine (6-MMP) simultaneously in red blood cells ( RBC ) of human.Methods The separation was performed on a Waters Xterra MS C 18 colum with mobile phase consisted of methanol -20 mmol · L-1 potas-sium dihydrogenphosphate -triethylamine ( pH adjusted to 3.2 using phosphoric acid ) ( 5∶95∶0.1 ) with the flow rate of 1.0 mL · min-1.The column temperature was 25 ℃.The detection wavelength was set at 348 nm for 6 -TGNs and 303 nm for 6 -MMP.Results The linear relationship range was 1.25 ×10 -2 -1.25 μg · mL-1 for 6 -TGNs ( r=0.999 6 ) and 0.04-5.00 μg· mL-1 for 6-MMP ( r=0.999 4 ).The lowest limit of quantification was 1.25 ×10 -2 μg · mL -1 for 6-TGNs and 0.04 μg · mL-1 for 6 -MMP.The RSDs of intra -day and inter-day of 6 -TGNs and 6 -MMP were less than 8.9%.The extraction recoveries were 60.99%-72.04% for 6-TGNs, 85.62%-95.13%for 6-MMP.The RSDs of 6-TGNs and 6-MMP stability were less than 6.08%.Conclusion The method is simple , rapid , sensitive , specific and stable , which could be applied to monitor thiopurine metabolites concentrations in red blood cells from inflammatory bowel disease and provide reference for personal medicine.关键词
硫嘌呤/6-硫鸟嘌呤核苷酸/6-甲基巯基嘌呤/高效液相色谱法/血药浓度Key words
thiopurine/6-thioguanine nucleotides/6-methlymercaptopurine/high performance liquid chromato-graphy/blood concentration分类
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朱霞,王雪丁,晁康,黄梓诚,郑洪,高翔,黄民..高效液相色谱法测定人红细胞6－硫鸟嘌呤核苷酸和6－甲基巯基嘌呤核苷酸的浓度[J].中国临床药理学杂志,2016,(3):257-260,4.基金项目
科技部“十二五”重大新药创制资助项目（2012ZX09506001－004）；国家自然科学基金项目 （）
