One of challenges for current glycomic analysis is that it often requires high quantity of initial sample,which is difficult to meet the needs of micro-sample analysis.In this study,a micro-sample N-glycan preparation technique(GPAT) was developed for the release,enrichment and analysis of N glycans in micro protein samples.By loading C18 and HILIC fillers separately in tips,a one stop platform was set up to perform protein digestion,N glycan release and enrichment in situ.Compared with other N-glycome analysis based on the filter aided sample preparation(FASP) strategy,N-glycome analysis with GPAT was able to detect 20 and 39 N glycans in 10 μg IgG and 10 μg HepG2 cell proteins,respectively,realizing almost the same identification effect with 90% decrease of the initial sample consumption.Otherwise,forty nine N glycans was also detected in 10 μg of urine protein extracted from the urine of 6 healthy people(3 males,3 females).Results show that GPAT realizes a simple and fast quantitative analysis for N-glycans in micro scale protein samples,and lays a foundation for further application of glycome researches.