Chemical Analysis and Biological Potential of Cotton Lavender Ethanolic Extract (Santolina chamaecyparissus L., Asteraceae)

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The authors studied the “Chemical analysis and biological potential of cotton lavender ethanolic extract (Santolina chamaecyparissus L., Asteraceae)”. Their approach is interesting but it has flaws that make this version unacceptable for publication. While I have no objection against publishing the data, I have some issues that need addressing:

1. Statistical methods in results are needed. Information about statistical methods obtained in the experiments must be provided and detailed in methods section.

2. In results, chemical analysis of plant extract needs to be checked by qualitative methods using HLPC/DAD (Authors do not explain how plant compounds were identified).

3. I am concerned about the poorly elements in the discussion; authors must be provided a good debate for results interpretation.

A few points:

L.30: Keywords should be in alphabetic order. Also, keywords serve to widen the opportunity to be retrieved from a database. To put words that already are into title and abstracts makes KW not useful. Please choose terms that are neither in the title nor in abstract.

Ls.34-35: Revise this sentence to eliminate rewordiness.

Ls.53-65: 

L.65: Also, a hypothesis in needed.

Ls.82-102: How do you identify the compounds? Did you do a qualitative analysis?

L.102: Define “d.e.”

Ls.137-139: What statistical methods do I use to analyze the data from each experiment?

Ls.172-178: The IC50 values should be estimated by probit analysis.

Ls.203-217: This information should be in results section (in annex) or delete.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors,

This study explores the chemical composition and biological potential of the ethanolic extract of cotton lavender (Santolina chamaecyparissus), with a focus on its relevance in phytotherapy and the antioxidant and antihyperglycemic activity of its main compounds. The introduction is well-structured, with objectives clearly presented and justified by relevant bibliographic references. Since you mention the primary compounds in the volatile oil, it would enhance clarity to also include the main antioxidant compounds referenced in the cited studies.

In the Materials and Methods section, there is no dedicated subchapter for reagents, which could improve clarity. Additionally, it is not evident if calibration curves were generated or which compounds were specifically calibrated. Within the subsection on statistical evaluation, specifying the statistical tests used would be beneficial. Currently, it appears that only the standard deviation was calculated.

Regarding the Results, there are some issues with the HPLC methods:

1. The chromatograms lack clarity; peaks are not integrated, and there are no associated retention times. Please consider addressing these issues.
2. It is unclear how the results were obtained. If calibration curves were made, the relevant data should be provided, including calibration curve equations, correlation coefficients, and the limit of quantification.

In the Discussion section:

1. The authors compare the polyphenol and flavonoid content of their cotton lavender extract with previous studies, noting that total polyphenol levels are higher in methanol extracts than in their ethanolic extract. However, beyond this quantitative comparison, much of the discussion centers on the findings from other studies, without a detailed interpretation of their own results. Although cited research highlights the importance of solvent choice and shows variable yields, the connection to the authors' findings is somewhat limited. The discussion could better integrate comparisons of their results on phenolic and flavonoid content, both qualitatively and quantitatively, with other studies.
2. Since molecular docking studies were conducted to investigate potential mechanisms of α-glucosidase inhibition, it would be advantageous to include structural comparisons with standard synthetic inhibitors (such as acarbose) or with natural antihyperglycemic compounds. Furthermore, the antihyperglycemic effects of both rutin and quercetin are already well-documented in numerous studies (a systematic review is available here: https://link.springer.com/article/10.1007/s13659-023-00424-w), suggesting that this aspect of the article may not offer new insights. While you highlight significant amounts of ferulic acid, further discussion of this compound would be valuable, as literature also reports antidiabetic effects for ferulic acid.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

ABSTRACT

Lines 13 – 16: The aim of this study............ specifically the content of phenolic and flavonoid compounds using high-performance liquid chromatography (HPLC), antioxidant activity.......

Change to: The aim of this study............ specifically the content of phenolic and flavonoid compounds using high-performance liquid chromatography (HPLC). Lines 16 18: The wording needs to be changed!

INTRODUCTION

Line 34: Aromatic plants were the first plants whose ..... Change to  Aromatic plants were the first whose .........

2. Materials and Methods   - Line 60

2.1. Plant material and extract preparation

Lines 72 – 73: ....of the e Institute of Field and Vegetable Crops Novi Sad (IFVCNS), ........

......Novi Sad (IFVCNS),.....  Question:  From where?

Line 75: University of 75 Novi Sad (Voucher No 2-1446).    Question:  From where?

RESULTS    -   Line 140

Lines 146 to 152: Based on the obtained results,...................... - rutin, in the examined extract was low.

Question: The paragraph does not have any discussion that explains, based on the chromatogram in Figure 2, how to understand the high ferulic acid content and the low rutin content!

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors have incorporated all suggestions and reviewer comments into the latest version, now the manuscript seems much clear. There are minor points to be corrected:

L.336: Statistical data should be in results section. Insert.

L.343: Statistical data should be in results section. Insert.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

I appreciate you making corrections and additions to the written material. However, in terms of chromatographic separation, interpreting the information presented in chromatograms is challenging. The chromatograms should have been enlarged to clearly see the separation, integration, baseline and resolution between the peaks. Peaks that are not properly separated, despite having large areas, are of limited relevance unless they represent compounds of interest. If they correspond to compounds of interest, such poorly resolved peaks can provide, at best, identification without providing additional analytical value.

Author Response

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Author Response File: Author Response.pdf