Analysis of Essential Oil of Salix babylonica Collected in Vietnam: Phytochemical Components and Antibacterial and Anticancer Activity

Round 1

Reviewer 1 Report

In the present report, Van and Phu have tried to delineate the chemical composition of essential oils from Salix babylonica. Furthermore, the authors have also tried to elucidate the anti-cancer and antimicrobial effects of the extracted oil using suitable in vitro models. Although the topic of this investigation is currently of importance nevertheless, I believe that this manuscript requires rigorous re-editing and several assays should be incorporated before it can be accepted for publication in this journal. The comments/suggestions are as follows:

1. The title of this manuscript should be changed. The title appears to be much from a master's thesis and is quite vague. 

2. The abstract needs to be rewritten by including the observations from the analysis. "Essential oils of this plant have demonstrated anti-bacterial activity against the tested bacteria, and anti-cancer against three cancer cell lines." This appears to be an unacceptable way of presenting the findings in an abstract.

3. Do the authors have previously read any research articles??? I am saying this because a three-paragraph introduction is a rare phenomenon in reviewing a research article. The authors are supposed to have given more information on the Plant being studied, its composition and pharmacological aspects of various parts of this plant should also be included in the Introduction. Previous reports of this plant or members of the family should also be stated to provide a comprehensive introduction and evidence for doing this study. The introduction should also state why the essential oils from this plant are important as anticancer or antimicrobial agents.

4. The manuscript is full of syntax and grammatical errors. I suggest the authors get this manuscript proofread for its grammar by a native English speaker and the proof of the same should be provided during the submission of the revised manuscript. 

5. Extraction process should be discussed in more detail for better reproducibility.

6. I believe that in the method section, heading no. 2.3 and 2.4 should be merged. Why the authors have used different headings?

7. In section 2.5, the strain number or accession number should be stated for providing more authenticity to the author's statement/s.

8. Standard name of the assay should be stated. Why the authors have chosen  25 g/mL, 50 g/mL, and 100 g/mL concentrations for disc diffusion assay?

9. Why the authors did not include the images of the disc diffusion assay in the manuscript? It should be included.

10. What do the authors understand by inhibitory assay?

11. In my little knowledge database, I have heard the term "adenocarcinomic" for the first time. I would like to be enlightened by the authors on what it means and whether is it suitable to use such terms in scientific communication.

12. If the authors have used MTT stain, technically the heading should have been cytotoxicity assessment of ....... OR simple MTT assay.

13. media-related specifications should have been included in the material section. 

14. The MTT section needs to be rewritten by including the steps. Readers will face some serious difficulties when they read this section. Information is missing regarding the number of each cancer cell used, wavelength and absorbance of MTT, for how long the cells were incubated with oils, etc.

15. Why the authors have used 12% FBS in the media?

16. The steps for calculating IC50 need to be stated. It is highly recommended that authors should first read some papers related to their work published in the journal before resubmitting their revision.

17. The information related to triplicate should be provided in the Statistical analysis section.

18. Why two-way Anova was chosen for statistical inferences?

19. Line no.94 what is RI? It should be explained for the first time.

20. Line no. 112 and 113 need to cite proper references.

21. Authors should discuss the pharmacological properties of α-Humulene, Pentacosane, Trans-Carvone oxide, Trans- 110 Caryophyllene. And it should also be linked with their present study.

22. Figure legends should be rewritten by properly giving sub numbers to the figures Fig.1(A) and ...... not like up and down as written in line numbers 137 and 138.

23.  table captions should be rewritten to provide complete information.

24. Data Availability Statement: Not applicable. Why this statement has been chosen?

The authors should get the manuscript re-edited by a professional English speaker and a certificate of the same should be provided if they are asked to resubmit this manuscript.

Author Response

REBUTTAL LETTER

          Firstly, our research team would like to thank the very useful comments of the Editors and Reviewers who contributed to our manuscript.

         We would like to send to the Editors and Reviewers the group's responses to the issues that the Editors and Reviewers have raised.

          Our team is looking forward to receiving more Editor's and Reviewers’s comments.

REVIEWER #1

1. The title of this manuscript should be changed. The title appears to be much from a master's thesis and is quite vague. 

Reply: We rewrite title as: “Analysis of essential oil of Salix babylonica collected in Vietnam: phytochemicals components and antibacterial and anticancer activity”

2. The abstract needs to be rewritten by including the observations from the analysis. "Essential oils of this plant have demonstrated anti-bacterial activity against the tested bacteria, and anti-cancer against three cancer cell lines."This appears to be an unacceptable way of presenting the findings in an abstract.

Reply: We revised in MS

3. Do the authors have previously read any research articles??? I am saying this because a three-paragraph introduction is a rare phenomenon in reviewing a research article. The authors are supposed to have given more information on the Plant being studied, its composition and pharmacological aspects of various parts of this plant should also be included in the Introduction. Previous reports of this plant or members of the family should also be stated to provide a comprehensive introduction and evidence for doing this study. The introduction should also state why the essential oils from this plant are important as anticancer or antimicrobial agents.

Reply: We revised in MS

4. The manuscript is full of syntax and grammatical errors. I suggest the authors get this manuscript proofread for its grammar by a native English speaker and the proof of the same should be provided during the submission of the revised manuscript. 

Reply: We revised in MS

5. Extraction process should be discussed in more detail for better reproducibility.

Reply: We added more detail for better reproducibility in Extraction process

6. I believe that in the method section, heading no. 2.3 and 2.4 should be merged. Why the authors have used different headings?

Reply: We revised in MS

7. In section 2.5, the strain number or accession number should be stated for providing more authenticity to the author's statement/s.

Reply: We revised in MS

8. Standard name of the assay should be stated. Why the authors have chosen  25 g/mL, 50 g/mL, and 100 g/mL concentrations for disc diffusion assay?

Reply: We revised in MS name of the assay as disc diffusion assay. We have chosen  25 g/mL, 50 g/mL, and 100 g/mL concentrations for disc diffusion assay because these concentrations usually chose for disc diffusion assay

9. Why the authors did not include the images of the disc diffusion assay in the manuscript? It should be included.

Reply: We added images of the disc diffusion assay in the manuscript

10. What do the authors understand by inhibitory assay?
11. In my little knowledge database, I have heard the term "adenocarcinomic" for the first time. I would like to be enlightened by the authors on what it means and whether is it suitable to use such terms in scientific communication.

Reply: Adenocarcinoma, it means that you have cancer in the glands that line your organs. This type of cancer can affect several different areas of your body, including the:

• Breast.
• Prostate.
• Pancreas.
• Esophagus.
• Colon/rectum.
• Stomach.
• Lungs.

12. If the authors have used MTT stain, technically the heading should have been cytotoxicity assessment of ....... OR simple MTT assay.

Reply: We revised as cytotoxicity assay

13. media-related specifications should have been included in the material section. 

Reply: We revised as in the material section

14. The MTT section needs to be rewritten by including the steps. Readers will face some serious difficulties when they read this section. Information is missing regarding the number of each cancer cell used, wavelength and absorbance of MTT, for how long the cells were incubated with oils, etc.

Reply: We revised in cytotoxicity assay

15. Why the authors have used 12% FBS in the media?

Reply: We revised as in the this protocol as 10% FBS in the media

16. The steps for calculating IC50 need to be stated. It is highly recommended that authors should first read some papers related to their work published in the journal before resubmitting their revision.

 Reply: We revised in cytotoxicity assay paragraph

17. The information related to triplicate should be provided in the Statistical analysis section.

Reply: We revised in cytotoxicity Statistical analysis section

18. Why two-way Anova was chosen for statistical inferences?

Reply: Because this is the method that allows for the most consistent statistical inference with studies of this type

19. Line no.94 what is RI? It should be explained for the first time.

 Reply: We revised in MS in the 2.3

20. Line no. 112 and 113 need to cite proper references.

Reply: We revised in MS

21. Authors should discuss the pharmacological properties of α-Humulene, Pentacosane, Trans-Carvone oxide, Trans- 110 Caryophyllene. And it should also be linked with their present study.

Reply: We revised in MS

22. Figure legends should be rewritten by properly giving sub numbers to the figures Fig.1(A) and ...... not like up and down as written in line numbers 137 and 138.

Reply: We revised in MS

23. Table captions should be rewritten to provide complete information.

Reply: We revised in MS

24. Data Availability Statement: Not applicable. Why this statement has been chosen?

Reply: Because the manuscript has provided enough images of the original data of the study. Please

Reviewer 2 Report

Authors conducted a comprehensive study on the chemical composition and potential biological activities of essential oils extracted from Salix babylonica in Vietnam. They utilized gas chromatography-mass spectrometry (GC-MS) to identify the chemical compounds present in the essential oils and discovered a significant number of compounds, with some being identified for the first time in this plant species. Additionally, the essential oils displayed promising antibacterial and anti-cancer activities against tested microorganisms and cancer cell lines.

Although the research concept is good, but the manuscript is not written well. The manuscript lack some technical part and it has many flaws. Some specific comments are given below.

Line 23: willowis??

Line 27: inner heat Leaves???.

Line 28: overturning, overturning??

Line 37-41: should be rewritten. Example: chemical composition and biological activity is repeated with three times in one para

Line 43: There is no methodology for how much bark Salix babylonica that they collected and used for oil extraction

Line 45: Faculty Of Biology ?why capitals for Of?

Line 48: Whether the oil containing moisture removed, if yes explain.

Line 50-56: The oven and MS parameters for compounds separations and detection are missing.  

Line 63: Is Pseudomonas aeruginosa-PA Gram-positive strain???

Line 72 : Why this step is important “1h, 4oC”.

Line 73: the test solutions replace with The test solutions!!

Line 81: what is MTT method?

Line 86: Logarit, it thinks it is Logarithm??

Line 94-98: repeated sentences

Line 99-107: as the author did not used the standard, is unacceptable to agree the compounds presence with mentioned quantity.

Line 117-118: what you want to specify about solvent extraction and steam distillation? Which one have more advantage in the level of metabolites extraction?

Why some of the comopound name in table 2 are red colour?

Line 150: what is DD value? In table 3.

In some places the scientific name of the plant is not ilalized. E.g., line 154.

Line 165: lần lượt [????

Authors conducted a comprehensive study on the chemical composition and potential biological activities of essential oils extracted from Salix babylonica in Vietnam. They utilized gas chromatography-mass spectrometry (GC-MS) to identify the chemical compounds present in the essential oils and discovered a significant number of compounds, with some being identified for the first time in this plant species. Additionally, the essential oils displayed promising antibacterial and anti-cancer activities against tested microorganisms and cancer cell lines.

Although the research concept is good, but the manuscript is not written well. The manuscript lack some technical part and it has many flaws. Some specific comments are given below.

Line 23: willowis??

Line 27: inner heat Leaves???.

Line 28: overturning, overturning??

Line 37-41: should be rewritten. Example: chemical composition and biological activity is repeated with three times in one para

Line 43: There is no methodology for how much bark Salix babylonica that they collected and used for oil extraction

Line 45: Faculty Of Biology ?why capitals for Of?

Line 48: Whether the oil containing moisture removed, if yes explain.

Line 50-56: The oven and MS parameters for compounds separations and detection are missing.  

Line 63: Is Pseudomonas aeruginosa-PA Gram-positive strain???

Line 72 : Why this step is important “1h, 4oC”.

Line 73: the test solutions replace with The test solutions!!

Line 81: what is MTT method?

Line 86: Logarit, it thinks it is Logarithm??

Line 94-98: repeated sentences

Line 99-107: as the author did not used the standard, is unacceptable to agree the compounds presence with mentioned quantity.

Line 117-118: what you want to specify about solvent extraction and steam distillation? Which one have more advantage in the level of metabolites extraction?

Why some of the comopound name in table 2 are red colour?

Line 150: what is DD value? In table 3.

In some places the scientific name of the plant is not ilalized. E.g., line 154.

Line 165: lần lượt [????

Author Response

REBUTTAL LETTER

          Firstly, our research team would like to thank the very useful comments of the Editors and Reviewers who contributed to our manuscript.

         We would like to send to the Editors and Reviewers the group's responses to the issues that the Editors and Reviewers have raised.

          Our team is looking forward to receiving more Editor's and Reviewers’s comments.

REVIEWER #2

Line 23: willowis??

Reply: We revised in MS as replacement by Salix babylonica 

Line 27: inner heat Leaves???.

 Reply: We revised in MS

Line 28: overturning, overturning??

 Reply: We revised in MS

Line 37-41: Should be rewritten. Example: chemical composition and biological activity is repeated with three times in one para

 Reply: We revised in MS

Line 43: There is no methodology for how much bark Salix babylonica that they collected and used for oil extraction

 Reply: We revised in MS

Line 45: Faculty Of Biology ?why capitals for Of?

 Reply: We revised in MS

Line 48: Whether the oil containing moisture removed, if yes explain.

Reply: We revised in MS, oil containing moisture removed by using Na₂SO₄ dry.

Line 50-56: The oven and MS parameters for compounds separations and detection are missing.  

Reply: We revised in MS, Please

Line 63: Is Pseudomonas aeruginosa-PA Gram-positive strain???

Reply: We revised in MS, Please

Line 72 : Why this step is important “1h, 4oC”.

 Reply: Because it takes time to diffuse to the agar plate

Line 73: the test solutions replace with The test solutions!!

Reply: We revised in MS, Please

Line 81: what is MTT method?

Reply: The MTT assay is a colorimetric assay for assessing cell metabolic activity. NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present. These enzymes are capable of reducing the tetrazolium dye MTT, which is chemically 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its insoluble formazan, which has a purple color.

Line 86: Logarit, it thinks it is Logarithm??

 Reply: We revised in MS, Please

Line 94-98: repeated sentences

Reply: We revised in MS, Please

Line 99-107: as the author did not used the standard, is unacceptable to agree the compounds presence with mentioned quantity.

Reply: We revised in MS, Please

Line 117-118: what you want to specify about solvent extraction and steam distillation? Which one have more advantage in the level of metabolites extraction?

Reply: We revised in MS, Please

Why some of the comopound name in table 2 are red colour?

Reply: We revised in MS, Please

Line 150: what is DD value? In table 3.

Reply: We revised in MS, Please

In some places the scientific name of the plant is not ilalized. E.g., line 154.

Reply: We revised in MS, Please

Line 165: lần lượt [????

Reply: We revised in MS, Please

Reviewer 3 Report

The manuscript "Identification of phytochemical compositions and inhibition activities of the essential oil of Salix babylonica" must be improved in all sessions.

The title needs improvement by specifying the type of inhibition and the location where the plant was gathered.

Here's a title suggestion:

Analysis of essential oil of Salix babylonica collected in Vietnam: phytochemicals components and antibacterial and anticancer activity

Abstracts

lines 11-13 must be remade and include the word respectively:

A total of 28 compounds and 31 compounds were present in leaves and bark essential oils, respectively. Among them, chemical compositions are the first time in this plant.

Could you provide some examples of compounds that are present in this plant for the first time, as mentioned in the abstract?

Introduction

This section requires further elaboration, including the mention of compounds associated with biological activity.

lines 26-28 It is necessary to provide references for every use.

Materials and Methods

Line 47. Considering replacing 3000ml with 3 L. If not, a space must be included between value and unit (3000 mL, instead of 3000ml) and unit milliliter as mL instead of ml.

Line 58. It might be more common to use Retention Time (RT) rather than Retention indices (RI). Perhaps a revision is worth considering.

Line 64 

Could you kindly remove the species abbreviation from the phrase?

"...gram-negative strains include Escherichia coli – E. coli, ..."

Lines 70

What is the initial concentration of microbial suspension? Spreading 0.1 mL of 10⁸ CFU/mL means a spread of 10⁷ CFU/mL?

Line 74

I believe there may be an error in the unit of essential oil concentration. The current measurements are 25 g/mL, 50 g/mL, and 100 g/mL. However, I think it should be 25 mg/mL instead. 

I noticed that ampicillin was used as a positive control at a concentration of 50 mg/mL. However, it seems like this may be too high. I suggest consulting the reference for guidance at https://doi.org/10.1016/j.cmi.2018.05.021

When using 50 μL of 50 mg/ml, the disc test yields 2.5 mg of ampicillin.

2.6. Inhibitory assay consider a change to:

2.6. Inhibitory assay against cancer cells 

Results and discussion

This section must be improved by rewriting.

Table 1 and Table 2. indicate the unit or RI

I find Table 3 to be quite confusing and suggest that it be reorganized. Specifically, I recommend that the unit of measurement for the diameter of inhibition growth be placed within the table itself, rather than in line 151. This will improve the clarity and organization of the information presented.

The table indicates the concentration of compounds instead of mass that was applied to the disc. Newly I highlighted the possible excessive mass of ampicillin and essential oil. DMSO indicates whether was used 50 μL in the disc or not.

Must be revised.

Author Response

REBUTTAL LETTER

          Firstly, our research team would like to thank the very useful comments of the Editors and Reviewers who contributed to our manuscript.

         We would like to send to the Editors and Reviewers the group's responses to the issues that the Editors and Reviewers have raised.

          Our team is looking forward to receiving more Editor's and Reviewers’s comments.

REVIEWER #3

Here's a title suggestion:

Analysis of essential oil of Salix babylonica collected in Vietnam: phytochemicals components and antibacterial and anticancer activity

Abstracts

lines 11-13 must be remade and include the word respectively:

A total of 28 compounds and 31 compounds were present in leaves and bark essential oils, respectively. Among them, chemical compositions are the first time in this plant.

Could you provide some examples of compounds that are present in this plant for the first time, as mentioned in the abstract?

Reply: We revised in MS, Please

Introduction

This section requires further elaboration, including the mention of compounds associated with biological activity.

Reply: We revised in MS, Please

lines 26-28 It is necessary to provide references for every use.

Reply: We revised in MS, Please

Materials and Methods

Line 47. Considering replacing 3000ml with 3 L. If not, a space must be included between value and unit (3000 mL, instead of 3000ml) and unit milliliter as mL instead of ml.

Reply: We revised in MS, Please

Line 58. It might be more common to use Retention Time (RT) rather than Retention indices (RI). Perhaps a revision is worth considering.

Reply: In studying the chemical composition of essential oils by GC/MS method, the RI value is more interested than the RT value, because RT f depends on the experimental conditions, and the RI value changes little. Please.

Line 64 

Could you kindly remove the species abbreviation from the phrase?

"...gram-negative strains include Escherichia coli – E. coli, ..."

Reply: We revised in MS, Please

Lines 70

What is the initial concentration of microbial suspension? Spreading 0.1 mL of 10⁸ CFU/mL means a spread of 10⁷ CFU/mL?

Reply: Initial concentration of microbial suspension as 10⁸ CFU/mL, we used 0,1 mL of 10⁸ CFU/mL for starting experiment.

Line 74

I believe there may be an error in the unit of essential oil concentration. The current measurements are 25 g/mL, 50 g/mL, and 100 g/mL. However, I think it should be 25 mg/mL instead. 

Reply: We revised in MS, unit of essential oil concentration is incorrect. Please

I noticed that ampicillin was used as a positive control at a concentration of 50 mg/mL. However, it seems like this may be too high. I suggest consulting the reference for guidance at https://doi.org/10.1016/j.cmi.2018.05.021

When using 50 μL of 50 mg/ml, the disc test yields 2.5 mg of ampicillin.

2.6. Inhibitory assay consider a change to:

2.6. Inhibitory assay against cancer cells 

Reply: We revised in MS

Results and discussion

This section must be improved by rewriting.

Table 1 and Table 2. indicate the unit or RI

I find Table 3 to be quite confusing and suggest that it be reorganized. Specifically, I recommend that the unit of measurement for the diameter of inhibition growth be placed within the table itself, rather than in line 151. This will improve the clarity and organization of the information presented.

The table indicates the concentration of compounds instead of mass that was applied to the disc. Newly I highlighted the possible excessive mass of ampicillin and essential oil. DMSO indicates whether was used 50 μL in the disc or not.

Reply: We revised in MS

Reviewer 4 Report

Reviewers Comments: Based on the given feedback, there are several concerns raised about the manuscript's suitability for publication in the journal. The reviewer acknowledges that the work includes characterization data but questions its novelty and contribution to the existing literature. They mention the abundance of reports on inhibitory activities of Salix babylonica essential oils from VietNam implying that this work may not bring significant new insights. But the author clam that the work is first time.  The reviewer also criticizes the organization and structure of the manuscript. They point out that the title does not align with the content and that the keywords fail to represent the work adequately. The introduction is deemed inadequate in conveying the paper's central idea. Furthermore, The antibacterial activity against the tested bacteria, and anti-cancer against three cancer cell lines are chosen randomly. Is there is any relation to chose the cancer cell line HepG2 cell (human 170 hepatocarcinoma) and MCF-7 cell (human breast carcinoma), and A549 cell line? Overall, the reviewer perceives the research data as scattered and finds the main idea of the paper unclear. They conclude that, in their opinion, the manuscript is not suitable for publication in the journal (Process). Based on this feedback, the authors can revise the manuscript by addressing the concerns raised, providing additional characterization data (cell viability images, Antibacterial activity images 1HNMR and C13NMR) as suggested, improving the organization and clarity of the paper, and emphasizing the novelty and significance of their work compared to the existing literature

English may be improved 

Author Response

Based on the given feedback, there are several concerns raised about the manuscript's suitability for publication in the journal. The reviewer acknowledges that the work includes characterization data but questions its novelty and contribution to the existing literature. They mention the abundance of reports on inhibitory activities of Salix babylonica essential oils from VietNam implying that this work may not bring significant new insights. But the author clam that the work is first time.  The reviewer also criticizes the organization and structure of the manuscript. They point out that the title does not align with the content and that the keywords fail to represent the work adequately. The introduction is deemed inadequate in conveying the paper's central idea. Furthermore, The antibacterial activity against the tested bacteria, and anti-cancer against three cancer cell lines are chosen randomly. Is there is any relation to chose the cancer cell line HepG2 cell (human 170 hepatocarcinoma) and MCF-7 cell (human breast carcinoma), and A549 cell line? Overall, the reviewer perceives the research data as scattered and finds the main idea of the paper unclear. They conclude that, in their opinion, the manuscript is not suitable for publication in the journal (Process). Based on this feedback, the authors can revise the manuscript by addressing the concerns raised, providing additional characterization data (cell viability images, Antibacterial activity images 1HNMR and C13NMR) as suggested, improving the organization and clarity of the paper, and emphasizing the novelty and significance of their work compared to the existing literature

 Reply: Thank you so much Sir/madam due to many useful comments for our MS. We revised in MS. Thank you so much

Reviewer 5 Report

Here are points of improvement for the ‘‘Identification of phytochemical compositions and inhibition activities of the essential oil of Salix babylonica’

For the abstract, clarify the sentence for instance, the phrase "Salix babylonica essential oils from VietNam for the first time" can be changed to "Salix babylonica essential oils from Vietnam."Combine similar points. For instance, stating that many compositions are identified for the first time can be mentioned once. Specify which strains of bacteria and cancer cell lines were used without getting overly detailed.

For introduction, Ensure consistent use of terminology. For instance, either use "Salix plants" throughout or the scientific name. Check for grammatical errors. "Salix plants are also used as a herb..." should be "Salix plants are also used as herbs..." Avoid colloquial terms like "thought out" – perhaps you meant "throughout."

For methodology, Clarify the extraction process. For instance, what's the purpose of distilling with sterile water for 6 hours? Ensure that the methods used are described in detail. For example, the conditions for GC/MS analysis should be presented clearly. For the antibacterial activity section, mention what is being measured: e.g., "The diameter of the inhibition zones around the wells was measured." Consistency in terms used: If you've used "essential oils" in one part, avoid switching to "test solutions" in another unless you've clarified they're the same. When mentioning standard methodologies (like MTT assay), it's beneficial to provide a brief overview so readers understand the purpose. Make sure references are used consistently. For instance, "[40]" appears in the methodology but it's unclear what this refers to without a references section.

For Results and Discussion Section, Line 188: Clearly state the differences in the chemical composition between the two provinces to provide a context for comparison. Line 195-199 & 220-226: Instead of listing the compounds in a paragraph format, consider presenting them in a table or a bullet-point format for improved readability. Line 200: The term "affordance of conditional growth" is not clear. Consider revising to clearly describe the influence of growth conditions on chemical composition. Line 206-213: The comparison with other studies is beneficial, but make sure to succinctly state the implications. Does this mean S. babylonica's composition changes based on geography? Line 227-230: While the point about the geographical influence on essential oil composition is reiterated, this can be merged with the earlier discussion on the same topic to avoid redundancy. The tables (e.g., Tables 1, 2, and 4) are extensive and might be more readable if any less relevant data is moved to supplementary information.

Dive deeper into the implications of the results. For example, what does the antibacterial and anti-cancer activity mean in the broader context of potential therapeutic uses of the oil?

In the conclusions Section, the conclusion succinctly restates the findings. However, it could delve a little deeper into the potential implications of these findings. State the practical applications or future studies that could stem from this research. Consider discussing the limitations of the study and any potential directions for future research.

In conclusion, the study offers valuable insights into the topic. However, by addressing the above points, the article can be made more impactful and reader-friendly.

Comments for author File: Comments.pdf

Minor editing of English language required

Author Response

For the abstract, clarify the sentence for instance, the phrase "Salix babylonica essential oils from VietNam for the first time" can be changed to "Salix babylonica essential oils from Vietnam."Combine similar points. For instance, stating that many compositions are identified for the first time can be mentioned once. Specify which strains of bacteria and cancer cell lines were used without getting overly detailed.

We revised in MS

For introduction, Ensure consistent use of terminology. For instance, either use "Salix plants" throughout or the scientific name. Check for grammatical errors. "Salix plants are also used as a herb..." should be "Salix plants are also used as herbs..." Avoid colloquial terms like "thought out" – perhaps you meant "throughout."

We revised in MS

For methodology, Clarify the extraction process. For instance, what's the purpose of distilling with sterile water for 6 hours? Ensure that the methods used are described in detail. For example, the conditions for GC/MS analysis should be presented clearly. For the antibacterial activity section, mention what is being measured: e.g., "The diameter of the inhibition zones around the wells was measured." Consistency in terms used: If you've used "essential oils" in one part, avoid switching to "test solutions" in another unless you've clarified they're the same. When mentioning standard methodologies (like MTT assay), it's beneficial to provide a brief overview so readers understand the purpose. Make sure references are used consistently. For instance, "[40]" appears in the methodology but it's unclear what this refers to without a references section.

We revised in MS

For Results and Discussion Section, Line 188: Clearly state the differences in the chemical composition between the two provinces to provide a context for comparison. Line 195-199 & 220-226: Instead of listing the compounds in a paragraph format, consider presenting them in a table or a bullet-point format for improved readability. Line 200: The term "affordance of conditional growth" is not clear. Consider revising to clearly describe the influence of growth conditions on chemical composition. Line 206-213: The comparison with other studies is beneficial, but make sure to succinctly state the implications. Does this mean S. babylonica's composition changes based on geography? Line 227-230: While the point about the geographical influence on essential oil composition is reiterated, this can be merged with the earlier discussion on the same topic to avoid redundancy. The tables (e.g., Tables 1, 2, and 4) are extensive and might be more readable if any less relevant data is moved to supplementary information.

Dive deeper into the implications of the results. For example, what does the antibacterial and anti-cancer activity mean in the broader context of potential therapeutic uses of the oil.

We revised in MS

In the conclusions Section, the conclusion succinctly restates the findings. However, it could delve a little deeper into the potential implications of these findings. State the practical applications or future studies that could stem from this research. Consider discussing the limitations of the study and any potential directions for future research.

 We revised in MS

In conclusion, the study offers valuable insights into the topic. However, by addressing the above points, the article can be made more impactful and reader-friendly.

 We revised in MS

Round 2

Reviewer 2 Report

NO

Author Response

Thank you so much Sir/madam