运用甘油磷酸氧化酶法测定冰冻红细胞解冻后残留甘油的含量  

Application of the GPO-PAP method for measuring the residual glycerol of deglycerolized RBC units

作　　者：夏红英[1] 石丽萍[1] 龚晓燕[1] 陈立[1] 杨玲玲[1] 任芙蓉[1] 

机构地区：[1]北京市红十字血液中心,100088

出　　处：《北京医学》2016年第4期353-356,共4页Beijing Medical Journal

摘　　要：目的建立测定冰冻红细胞解冻洗涤后残留甘油含量的酶学方法测定工艺。方法进行甘油磷酸氧化酶法的线性范围测定、重复性、准确性（回收试验）、游离血红蛋白干扰试验,并比较不同样本处理方法（钨酸钠裂解法、三氯醋酸裂解法及制备后2 h直接取上清液法）对酶法检测甘油的影响;进行系列浓度甘油红细胞酶法测定结果与传统过碘酸钠滴定法测定结果比对。结果用GPO-PAP法检测甘油浓度在0.05~2.0 g/L时有较好的线性关系,r2=0.9994,Y=0.942 X＋0.0239;甘油含量为0.2 g/L、1.0 g/L的样本,批内CV值分别为2.68%、0.81%（n=20）,日间CV值分别为4.59%、4.37%（n=20）;甘油浓度从0.29~2.09 g/L间其回收率97.1%~100.8%,样本的平均回收率为99.0%;10 g/L游离血红蛋白对GPO-PAP法检测结果无干扰;3种样本处理方法所测得的系列浓度甘油红细胞样品及解冻去甘油红细胞制品（n=18）酶法测定结果一致（P〉0.05）;系列浓度甘油红细胞测定结果显示酶法测定比滴定法更准确,滴定法测定值偏高。结论甘油磷酸氧化酶法可用于冰冻解冻去甘油红细胞制品中残留甘油含量的测定,方法简便、快速、准确、重复性好。3种样本处理方法对GPO-PAP法检测甘油含量无影响。Objective To establish the procedure of an enzymatic method for measuring the residual glycerol concentration of cryopreserved, deglycerolized RBC units, using the partial detection principle of the enzymatic method（GPOPAP） for triglyceride. Methods Validation of the glycerol phosphate oxidase method（GPO-PAP） consisted of determining the linearity, precision and accuracy（recovery experiments）experiments and free- hemoglobin interference tests. The effects of different sampling process（sodium wolframate treatment, trichloroacetic acid treatment and directly sampling supernatant from deglycerolized RBC units 2h after preparation） on the accuracy of glycerol were compared. Results of series concentrations of glycerol of RBC measured by enzymatic method were compared with results by the traditional sodium periodate titration method. Results The GPO-PAP assay kit was found to be linear in the range of 0.05~2.0 g/L glycerol, with r2= 0.9994, Y=0.942 X ＋ 0.0239; Within-run imprecision was 0.74%（0.2 g/L glycerol） and 0.81%（1.0 g/L glycerol）（n=20）, and day-to-day imprecision was 4.59% and 4.37 %（n=20）. The recovery rate was from 97.1% to 100.8% when the Glycerin concentration was from 0.29 to 2.09 g/L, with the mean recovery 99.0%. 10 g/L free-hemoglobin did not interfere the assay. Different sampling processes had no effect on the glycerol results of washed RBC with series glycerol concentration and of cryopreserved, deglycerolized RBC units（n=18）（P 0.05）. Results Series concentrations of glycerol of RBC showed that enzymatic method is more accurate than titration method（Ttitration method gave higher values than its real content）. Conclusion GPO-PAP assay is a simple, rapid, accurate, precise method for measuring the residual glycerol concentration of cryopreserved, deglycerolized RBC units. Three sampling treatments do not affect the assays.

关 键 词：甘油 酶法 红细胞裂解液 解冻去甘油红细胞 

分 类 号：R446.11[医药卫生—诊断学]