Abstract: Objective To study the effect of different vacuum blood sampling tubes on whole blood glycohemoglobin（HbAlc） detected by high performance liquid chromatography（HPLC）. Methods Venous blood samples were collected from 40 outpatients with 4 kinds of vacuum blood sampling tubes.The samples were frozen and then thawed for measuring HbA1c with the RIO-RAD VariantⅡHPLC analyzer. Results The HbA1c was 7.88%±1.2%,7.96%±1.2%,7.91%±1.2%,and 7.95%±1.2% in samples collected with purple hat（K2-EDTA）,gray hat（sodium fluoride/heparin sodium）,green hat（heparin lithium） and blue hat（sodium citrate） respectively,which was not significantly different from those collected with vacuum blood sampling tubes（P＞0.05）.After 3 times of freezing and thawing,the HbA1c in samples collected with purple,gray and green hats was significantly different from those collected with different blood sampling tubes（P＜0.05）. Conclusion Vacuum blood sampling tubes can be in collecting whole blood HbA1c samples.Over three times of freezing and thawing of the sample can influence the detection of HbA1c.