中文摘要：

目的：探讨离子交换高压液相色谱法（HPLC）在检测相关定性定量指标时的干扰因素,提高血红蛋白病的确诊率。方法：应用D-10Dual Program仪器的地中海贫血（地贫）模式,利用HPLC测定人全血中HbA2/HbF的百分含量,分析洗脱时间、峰形及其他影响测值的因素。结果：1样本前处理的影响：提取样品的血红蛋白后进行上样分析,发现血红蛋白抽提液（1∶200的比例稀释）的上样分析效果显著优于全血（1∶300的比例稀释）直接上样,能更好地使地贫模式的总面积控制在1 000 000～4 000 000μvolt·s,同时可避免进样管堵塞事件的发生。2年龄因素的影响：新生儿HbF的百分含量高,可致峰值过饱和而使测值不准确或测不出,需用碱变性试验进行验证。3疾病类型的影响：重症β地贫、HbF持续增高症等血红蛋白病也以HbF明显增高为主,同样可出现前述峰值过饱和现象,而致HbF测值不准确或测不出,需倍比稀释或梯度稀释样本再行测定或行碱变性法进行验证。4异常血红蛋白病峰形观察：异常血红蛋白病的峰形图会有特征性变化,比如血红蛋白H病会在洗脱时间0.37min左右出现unknown值,血红蛋白S病会出现S-Window值,血红蛋白C病等其他异常血红蛋白病会出现C-Window值等。结论：样本稀释度的高低会影响地贫模式下的峰形扫描总面积,从而影响HbA2/HbF测定值的准确性;对新生儿和重症地贫等样本并行碱变性试验可避免误诊和漏诊;对异常血红蛋白病峰形的观察为进行地贫基因分析作了很好的提示。

英文摘要：

Objective：To investigate interference factors in qualitative and quantitative analysis by high pressure liquid chromatography（HPLC）and to improve the diagnosis rate for hemoglobinopathy.Method：Using the D-10 Dual Program Instrument,HbA2/HbF in whole blood was detected by HPLC method in thalassemia mode.The elution time,the peak line diagram and other interfering factors were analyzed.Result：1Influence of the sample pretreatment：The results of using the extraction of hemoglobin from the sample and dilution in the ration of1∶200as the sample was much better than directly using the whole blood which was diluted in the ration of 1∶300automated by the D-10 Dual Program.The extraction of hemoglobin diluted in 1∶200could precisely control the test area from 1,000,000 to 4,000,000μvolt·s and avoid pipe plug.2Age of the patients：Neonatal fetal hemoglobin content was high in newborns,which can result in HbF over saturation and interference measurement.In this condition,alkaline denaturation was necessary for verification testing.3Disease types：Hemoglobinopathies such as the severeβ-thalassemia or fetal hemoglobin continued to increase syndrome also have high HbF content,which can result in HbF oversaturation and interference measurement.Samples from these patients also need multiple proportion dilution or alkaline denaturation.4Analysis the peak line diagram of the abnormal hemoglobinopathies：Abnormal hemoglobinopathies had unusual peak line in the diagram.For example,the HbH symptom had the unknown value in 0.37 min elution time,the HbS symptom had the value of S-Windowin,the HbC symptom and other abnormal hemoglobinopathies had the value of C-Windowin.Conclusion：The different ration of dilution of sample scan changes the test area in the program,which also affects the exact detection value of HbA2/HbF.Using alkaline denaturation to detect HbF in newborns and severe thalassemia patients can decrease misdiagnosis rate.Analysis the peak line diagram of the abnormal hemoglobinpathies could provid