离子对反相HPLC测定小鼠骨髓BFU-E、CFU-E中PRPP合成酶活性  

Estimation of Phosphoribosylpyrophosphate(PRPP) Synthetase Activity in BFU-Es and CFU-Es from Mice Bone Marrows by Ion pair rHPLC

作　　者：刘立鹏[1] 卢义钦[1] 刘俊凡[1] 

机构地区：[1]湖南医科大学血液生化研究室,中国湖南长沙410078

出　　处：《生命科学研究》2000年第2期124-129,共6页Life Science Research

基　　金：国家自然科学基金资助项目!(39670908)

摘　　要：采用离子对反相高效液相色谱法 ( IPr HPLC)的单液等度洗脱 ,测定了小鼠骨髓红系爆增性集落形成单位 ( BFU- Es)与红系集落形成单位 ( CFU- Es)的 PRPP合成酶活性 .方法学研究表明 ,加入离子对试剂硫酸氢四丁基铵 ( TBAHS)后 ,ADP谱峰分离完全 ,其反应增加量易于计算 .本法的批内变异系数平均值为 - 2 .30 %～ - 1 .0 6%与 1 .0 6%～ 2 .30 % ,平均相对偏差为 0 .81 %～ 1 .74% ,回收率为 93.9%～ 98.5% ,能达到操作简便、灵敏和准确的要求 .采用本法测得 2 0只正常昆明小鼠的 BFU- E和 CFU- E中该酶活性各为 ( 0 .563± 0 .0 2 7)μmol/( min·g· m L- 1)与 ( 0 .41 6± 0 .0 2 6) μmol/( min·g·m L- 1) .The PRPP synthetase activity in BFU Es and CFU Es from bone marrows of mice was estimated by ion pair reversed phase HPLC (IPrHPLC),using isocratic elution. The results indicated that ADP peaks were completely separated from those of AMP and ATP after addition of the ion pair reagent tetrabutylammonium hydrogen sulfate (TBAHS),therefore the increments of the former could be easily calculated.By this method,the intrabatch coefficient of variation was -2.30%～-1.06% and 1.06%～ 2.30% on average;the mean relative deviation was 0.81%～1.74%,with a recovery rate of 93.9%～98.5%,fulfilling the requirements of simplicity, reproducibility and accuracy. The mean values of PRPP synthetase activities in BFU Es and CFU Es from bone marrows of 20 healthy Kunming mice were (0.563±0.027) μmol/(min·g·mL -1 ) and (0.416±0.026) μmol/(min·g·mL -1 ) respectively.

关 键 词：红系祖细胞 PRPP合成酶 IPrHPLC 核苷酸代谢 

分 类 号：Q559.103[生物学—生物化学] Q591.3