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.