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The current quality control(QC)of Radix Isatidis(R.Isatidis)by the fingerprint analysis is dominantly relied on reversed-phase liquid chromatography(RPLC),which may fail to provide sufficient information of many polar components.In this study,hydrophilic interaction liquid chromatography(HILIC)was introduced to promote the QC of R.Isatidis.We first analyzed 16 batches of samples with ultra-performance liquid chromatography tandem mass spectrometer,comparing traditional RPLC with HILIC.The resulting data of the two fingerprints were analyzed using analysis of similarity and retention behavior,showing that HILIC can increase the fmgerprint coverage for RPLC analysis.Then,an anti-viral assay ofR.isatidis was conducted to evaluate its efficacy.The fingerprint-efficacy relationships between the two fingerprints and anti-viral activity were investigated with principal component regression analysis.The results showed that the anti-viral activity correlated better with HILIC fingerprint(R=0.9638,P=0.016)than with RPLC fingerprint(R=0.7804,P=0.0003).This study indicated that HILIC fingerprint could better reflect the quality ofR.Istidis,which might be used to promote the QC of herbal medicines.