Study on Chemical Diversity, Antioxidant and Antibacterial Activities, and HaCaT Cytotoxicity of Camphora tenuipilis (a Traditional Aromatic Plant from Xishuangbanna)

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1) In its current way, this manuscript is preliminary. I encourage the authors to perform more experiments with the bioactive compounds found in the essential oils and perform pharmacodynamic studies evaluating the molecular mechanism of action. This manuscript should present the results, not subsequent works.

2) The aim of the study in the abstract section is not clear.

3) Lines 24-26. "This study provides the first scientific evidence supporting the traditional use of C. tenuipilis in "Duo Sheng," highlighting its potential to extend food shelf life and reduce consumption risks." Lines 454-456. "To assess safety, we evaluated the cytotoxicity of these essential oils on human keratinocytes (HaCaT), finding IC50 values ranging from 0.163 to 0.847 mg/mL, indicating low toxicity at certain concentrations." The cytotoxicity of these essential oils on non-tumorigenic cells is high. The IC50 values are low. Are the authors sure about this statement? Acute toxicity tests in rodents are necessary in this work.

4) The introduction section lacks ethnomedicinal information about this plant species.

5) Is there a clear justification for performing the biological assays with the essential oils of this plant species?

6) A reference drug (positive control) is missing in all the biological assays. How can the authors compare the potential of the essential oils without this control?

7) A vehicle group is missing in the cytotoxicity assay.

8) Section 4.1 should indicate a voucher number.

9) What was the yield of extraction for each essential oil?

10) The cytotoxicity of these essential oils should be evaluated on other non-tumorigenic cell lines.

11) The format of some references is not homogeneous (e.g., 26).

12) The conclusion section needs to be rewritten.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

Accept in current form

Author Response

Reviewer 2 has recommended "Accept in current form", thus there are no revision comments here.

Reviewer 3 Report

Comments and Suggestions for Authors

The article comprehensively discusses the chemical diversity, antioxidant and antibacterial activities, and cytotoxicity of essential oils extracted from Camphora tenuipilis leaves in HaCaT cells. The study provides scientific support for both traditional use (the "Duo Sheng" dish) and potential food, cosmetic, and pharmaceutical applications. The identification of five different chemotypes using GC-MS analysis and their correlation with biological activities make the study unique. The article is generally well-structured, methodologically detailed, and the results are discussed with the literature. However, there are some methodological, linguistic, and structural shortcomings.

1- While the introduction is comprehensive, some statements are repetitive and the focus shifts. The problem definition, hypothesis, and research gap need to be highlighted more clearly.

2- Only one person (Mr. Zhou Shishun) is cited in the identification of the plant material, but no voucher number/herbarium reference is provided.

3- Information on positive controls (e.g., standard antibiotics) is missing in antibacterial tests.

4- The concentration ranges and number of replicates used in cytotoxicity experiments should be more clearly specified.

5- Tables and figures contain ample information, but in some sections, the graphic and table descriptions are insufficient (e.g., Figure 2 and Figure 4).

6- Antibacterial test results (MIC, MBC) need more robust comparative interpretations.

7- The discussion section is long, but some points are repetitive. Comparisons with the literature should be more critical, not only supporting but also explaining the differences.

8- The risk-size assessment of cytotoxicity findings in terms of potential applications should be more detailed.

9- Some sentences are long and complex; the language should be more fluent.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 4 Report

Comments and Suggestions for Authors

The authors reported a study of the chemical diversity and biological activities investigation of Camphora tenuipilis. A few points need to be addressed for clarification.

1. Page 3 Line 92. Introduction. What is the traditional seasoning method?

2. 2.1. Chemical composition analysis of five C. tenuipilis essential oils. The authors categorized 5 chemotypes. However, there are also only 5 sample in this study. In order to categorize certain chemotypes, there should be more extracts to compare the similarities and then classify different types.

3. Page 3 Line 122. “the essential oils BX07, FH01, FH07, YC02, and YC04 can be categorized as five distinct chemotypes of C. tenuipilis leaf essential oils: linalool-type, citral-type, geraniol-type, elemicin-type, and methyl cinnamate-type, respectively.” BX07, FH01, FH07, YC02, and YC04 represent different Camphora tenuipilis samples. The sentence should be revised to avoid confusion. Same revision should be made to Table 2 title.

4. Figure 1 and Figure S1. The resolution should be increased. What do BX07-1, BX07-2, BX07-3 represent? The information should be added to the method section if the same samples were analyzed in triplicate.

5. Table 2. Full name and abbreviation of TP may also be mentioned in the legend. The meaning for letters a, b, c, d, e should also be added. Same applies to Table 3.
z
6. Page 8 Line 194. “Citral and linalool, components of the essential oils, demonstrated effective antibacterial properties against the four bacteria.” A reference should be added if this statement is supported by other articles, or a demonstration will be needed regarding how these two compounds were identified as key components to these activates.

7. Page 12 Line 329. “Dried samples from five species of C. tenuipilis leaves” All five samples are from the same species. The sentence should be revised, the manuscript may be thoroughly checked to avoid similar confusions.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

In its current way, this manuscript is preliminary. I encourage the authors to perform more experiments with the bioactive compounds found in the essential oils and perform pharmacodynamic studies evaluating the molecular mechanism of action. This manuscript should present the results, not subsequent works. The authors did not perform these corrections. This information is essential for improving the manuscript.

The authors did not perform any additional toxicity study to guarantee the safe use of the plant extract. If this essential oil is used in traditional medicine, it does not guarantee its lack of toxicity.

There is no clear justification for performing the biological assays with the essential oils of this plant species. The authors did not show ethnomedicinal evidence that supports the use of this plant species.

I could not locate any reference drug in the cytotoxic or antioxidant assays.

There is no vehicle group in the cytotoxic and antioxidant assays.

 

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 4 Report

Comments and Suggestions for Authors

Regarding the response to Q2. “Chemical composition analysis of five C. tenuipilis essential oils. The authors categorized 5 chemotypes. However, there are also only 5 sample in this study. In order to categorize certain chemotypes, there should be more extracts to compare the similarities and then classify different types.”

It is still confusing since all the tables and figures only list 5 samples instead of 15 samples. The response to Q3 also indicated that there are five essential oil samples. Section 4.2 need to be revised to clarify that there are in total of 15 samples, not 5 samples with triplicates.

Comments on the Quality of English Language

4.2. Isolation of essential oils need to be revised.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Round 3

Reviewer 1 Report

Comments and Suggestions for Authors

In its current way, this manuscript is preliminary. I encourage the authors to perform more experiments with the bioactive compounds found in the essential oils and perform pharmacodynamic studies evaluating the molecular mechanism of action. This manuscript should present the results, not subsequent works. Once more again, the authors did not perform these corrections. This information is essential for improving the manuscript.

Figures 2 and 4 do not indicate a vehicle group or a reference drug.

If Camphora tenuipilis is an edible plant, the authors should perform experiments and analysis related to its nutritional content. There is no need to perform other biological experiments with an edible plant.

Once again, the authors did not conduct any additional toxicity studies to ensure the safe use of the plant extract.

Author Response

Please see the attachment.

Author Response File: Author Response.docx