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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 erythrocytic burst forming units (BFU-Es) and erythrocytic colony forming units (CFU-Es) in mouse bone marrow were measured by single-liquid isocratic elution using ion-pair reversed-phase high performance liquid chromatography ) Of the PRPP synthase activity methodological studies showed that the addition of ion-pair reagent tetrabutylammonium hydrogen sulfate (TBAHS), ADP peak separation is complete, the increase in the amount of reaction easy to calculate.The average variation coefficient of this method was - 2.30% ~ - 1.06% and 1.06% ~ 2.30%, the average relative deviation was 0.81% ~ 1.74%, the recovery rate was 93.9% ~ 98.5%, to achieve the operation Simple, sensitive and accurate.Using this method, the enzyme activity of BFU-E and CFU-E in 20 normal Kunming mice was (0.563 ± 0.027) μmol / (min · g · m L- 1) and (0.41 6 ± 0 .0 26) μmol / (min · g · m L -1).