Abstract: This study employed UHPLC-QTOF-MS based metabolomics to investigate the effect of different extraction solvents and chromatography separation modes on the metabolomic analysis of mango pulp, providing a comprehensive and high-throughput analytical method mango pulp. Four extraction solvents—H₂O, H₂O with 0.1% acetic acid (H₂O+A), 50% MeOH (Me), and 50% MeOH with 0.1% acetic acid (Me+A)—were utilized alongside two chromatography modes, HILIC and RPLC. The results indicated that the addition of 0.1% acetic acid significantly enhanced the quantity and peak area of MS1 and MS2 particularly in the high-response region (peak area>10⁷). Annotation results revealed that the number of annotated components in the H₂O+A group was 1.39 times greater than that of the H₂O group, and the Me+A group annotated 1.15 times more than that of the Me group. Additionally, MS2 spectra from the H₂O+A and Me+A groups were cleaner, with less noise interference, leading to more accurate annotation results. The combination of (H₂O+A)+HILIC and (Me+A)+RPLC exhibited strong complementarity. This strategy resulted in the annotation of 311 metabolites across mango pulp samples, nearly doubling quantity and expanding the polarity range by 2.4 times compared to the conventional method of (Me+A)+RPLC alone. Combining (H₂O+A)+HILIC with (Me+A)+RPLC significantly enhanced metabolite coverage and annotation accuracy. It provides a more comprehensive analytical method for the analysis of components in fruit and vegetable products such as mango pulp.