Abstract: Objective:To establish a green and environmentally friendly HPLC fingerprint analysis method for Morchella. Methods:An ionic liquid(1-octyl-3-methylimidazolium hexafluorophosphate)-water solution was as extracted-solution. The effects of experimental conditions(the type of ionic liquid,ionic liquid carbon chain length,ionic liquid mass fraction of solution,extraction time,and material-liquid ratio)on the extraction amount were optimized through the single factor test,the final extraction condition was established as 40%OMIMPF₆ aqueous solution(material-liquid ratio 1:10)wet grinding sample for 10 min. Then green HPLC was used to analyze Morchella samples. The separation was performed on a Agilent ZORBAX SB-AQ C₁₈ column(4.6 mm×150 mm,5 μm)with gradient elution by 0.2% acetic acid-ethanol as a green mobile phase system at 1.0 mL/min,column temperature was 40℃,and detection wavelength was at 260 nm. The "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition)"was used to evaluate the similarity of 20 batches of samples(14 batches of Morchella sextelata,6 batches of various edible medicinal fungi). And the cluster analysis and principal component analysis were performed on the samples by SPSS software. Results:HPLC fingerprint analysis approach of Morchella was established,calibrating 10 common peaks,and 5 nucleoside components(uracil,cytidine,uridine,guanosine,and adenosine)were identified. The similarity of 14 batches Morchella sextelata was above 0.9,and the similarity of the other 6 batches samples was among 0.25~0.65. The clustering and principal component analysis also showed that the chemical compositions between Morchella and other edible medicinal fungi were obviously different under this experimental condition. Conclusion:A convenient,green and rapid HPLC fingerprint analysis approach for Morchella was established,which would help to improve the comprehensive quality evaluation level of Morchella.