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Thanks for addressing all comments!
[# PeerJ Staff Note - this decision was reviewed and approved by Sonia Oliveira , a PeerJ Section Editor covering this Section #]
Please address all reviewer´s comments.
**PeerJ Staff Note:** Please ensure that all review and editorial comments are addressed in a response letter and that any edits or clarifications mentioned in the letter are also inserted into the revised manuscript where appropriate.
**Language Note:** The review process has identified that the English language must be improved. PeerJ can provide language editing services - please contact us at [email protected] for pricing (be sure to provide your manuscript number and title). Alternatively, you should make your own arrangements to improve the language quality and provide details in your response letter. – PeerJ Staff
The study is an original and comprehensive study. The English grammar should be checked again. Discussion is very poor; it should with compared with the literature.
Experimental design is good. However, standard deviation may be added with more replication. Methods are sophisticated.
Validity may be enriched with more replications.
This study presents a detailed untargeted metabolomics approach, featuring extensive compound annotation and the validation of six Q-markers for biological relevance. The integration of statistical analysis, bioactivity screening, and structural considerations enhances the manuscript. However, several methodological and reporting aspects require clarification to ensure the robustness, reproducibility, and biological relevance of the proposed markers.
The English is clear, and the literature cited is sufficient. However, not all raw data have been shared. It is recommended to include the raw data as a supplementary file or to deposit the raw LC-MS/MS data and associated metadata in a recognized public repository such as MetaboLights, GNPS, or the Metabolomics Workbench.
Additionally, for Figure 6—Heatmap of the 6 Q-markers—the figure caption should be more descriptive, clearly specifying what the values in the heatmap represent.
The selection of Q-markers based on VIP scores, p-values, structural uniqueness, and traditional bioactivity is reasonable. However, this approach may overlook other biologically relevant metabolites. The authors should justify why these specific criteria were chosen and clarify whether additional metabolites were considered or excluded, and why independent validation or functional assays were not performed.
The manuscript should include the identification confidence levels (e.g., MSI levels) of the six Q-markers.
The author did not provide details on peak picking and missing value handling.
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