血红蛋白异常对高效液相色谱法检测糖化血红蛋白的影响

Effect of hemoglobin variant on assay of glycohemoglobin HbA1c by high performance liquid chromatography

目的探讨血红蛋白(Hb)异常对高效液相色法谱检测糖化血红蛋白(HbA1c)的影响。方法使用国内常用的硼酸盐亲和层析高效液相色谱法(PDQ系统)及离子交换层析高效液相色谱法(D-10系统)进行含有不同(梯度)浓度HbF与正常人血液混合的标本的干扰实验及检测40例Hb异常的临床标本中的HbA1c。结果在干扰实验中,D-10系统在HbF在浓度超过40%时,可能无法获得检测结果,当HbF浓度在10%～40%时,结果高于实际值,HbF在浓度低于10%时,结果与实际值相似;PDQ系统在HbF浓度超过40%时,结果可能略低于实际值,HbF浓度低于40%时,可以获得好的结果。在40例Hb异常的临床标本中,D-10系统检测结果明显高于PDQ系统,差异具有统计学意义(P<0.05),其中有5例Hb异常总量超过35%的标本在D-10系统中无法获得检测结果,而PDQ检测获得可接受的结果;Hb异常总量低于10%时,两种方法检测结果有较好的一致性(P>0.05)。结论硼酸盐亲和层析抗Hb异常的干扰能力优于离子交换层析。

Objective To discuss the effect of hemoglobin variant on the assay of glycohemoglobin HbAlc by high performance liquid chromatography （HPLC）. Methods The results of hemoglobin HbAlc from 40 samples with hemoglobin variant were obtained using the Primus PDQ （based on Boronate -affinity chromatography） and the Bio-Rad D-10 （based on Ion-exchange chromatography） ; and an interference test was designed to detect the interference of HbF （in different concentration） in the assay of HbAlc by above-mentioned two HPLC. Results In the interference test, the results of HbA1c maybe not obtained by D-10, while they could be obtained acceptable, but under the true,by PDQ,when the concentration of HbF exceeded 40%. When the concentration of HbF was between 10% and 40% ,the results from D-10 exceeded the true,and the results from PDQ were nearly the true. When the concentration of HbF was under 10% ,the results from the two methods were in close agreement. In the 40 clinical samples with hemoglobin variant,the results from D-10 exceeded obviously the ones from PDQ,and they showed there was significant differences（P〈0. 05）between the two methods; and five samples, which were concentration of variant were above 40 %, were not obtained in the D-10, but could be obtained in PDQ. When the concentration of variant was under 10% ,the results from the two methods were in close agreement（P〉0.05）. Conclusion The capability of antiinterference to variant of boronate affinity chromatography is excelled that of ion-exchange chromatography.