Sustainable Characterization of Some Extracts of Origanum vulgare L. and Biosafety Evaluation Using Allium cepa Assay
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe article focuses on the study of Origanum vulgare extracts to determine their potential implications for Romanian traditional medicine. The authors utilized a sustainable Allium cepa assay method which is sensitive to cytotoxic and genotoxic substances. The authors have identified 7 physicochemical parameters, the analytical results are well-presented. The statistical methods (Pearson correlation coefficient matrix of the measured parameters ; Schoeller-Berkaloff diagram ; ) used are relevant.
It is important to highlight the added value of their study compared to similar studies already published (Bulgarian study, reference 34). The HPTLC technique could have been described in greater detail, in particular in relation to standards chosen from the literature or a broader reference database.
The article is written clearly with very few typos (example line 192 : apig-191 enin-7-O-glucoisde). The cited publications have been carefully selected to highlight the methods used and provide an overview of similar studies, although there is a need to standardize the formatting of references (year in bold, pages, etc..)
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to review this manuscript, for your comments and suggestions.
Please find the detailed responses in the attached document and the corresponding revisions/corrections highlighted in the re-submitted manuscript.
Best regards!
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe comments are as follows:
1. The title should be changed to better represent the aim of the work and the results obtained.
2. Statistical analysis should be included in Table 1 and 4.
3. The authors should improve the discussion and comparison with scientific literature.
4. Moisture content of dried sample material has to be included in subsection 3.1. Was the sample material ground?
5. Calibration curve and correlation factor should be provided for TPC.
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to review this manuscript, for your comments and suggestions.
Please find the detailed responses in the attached document and the corresponding revisions/corrections highlighted in the re-submitted manuscript.
Best regards!
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsIn Table 1, the results presented have many decimals. Are they necessary? How many extracts’ replicates were made? Is it possible to do a statistical analysis, to be sure about the differences among the samples?? Analysis of variance (ANOVA)? The same question for Table 3.
In the footnote of Table 1: *Calculated as the sum of the absorbance at 420 (yellow), 520 (red), and 620 (blue) nm (values registered for each absorbance not shown). Why in Material and Methods, there is the following sentence “As well, by scanning the oregano extracts in a range of 190-440 nm, the fingerprint of the UV spectra was recorded.”?
The procedure is usually used for the chromatic characteristics of wine [50] and recently extended to the color evaluation of different beverages [50,51]. After reading these references, the samples were red, I am not sure if this is appropriate to use this method for doing the color of oregano extracts.
Figure 1 can be taken off as well as the sentence: In addition, the logarithmic representation of these data (Figure 1) allows a better visual comparison, highlighting the relationship between the seven indicators selected for monitoring the influence of the extraction conditions on the chemical profile of the extracts, as well as the therapeutic potential.. It does not provide important information.
The results observed in section 2.1.1. need to be discussed. This is absent; the authors only describe the results, without any type of discussion.
In the Conclusion, the authors have written:
The present study has allowed the in vivo evaluation of cytotoxicity and genotoxicity potential of different oregano extracts using Allium cepa assay, a fast and easy-to-handle method, with reliable results, minimal cost, and environmentally friendly.
The authors allow me to disagree with some points of the sentence: a fast and easy-to-handle method, because:
onion bulbs were placed in tap water for 72 hours. The next day, a series of 3 bulbs were maintained in the four oregano extracts as such, without any dilution in controlled conditions for 24 hours in the dark, at 22 °C…. Then: To make the cytogenetic preparations, the roots were subjected to processing steps. Roots were fixed in Farmer's solution, for 18 hours in the refrigerator…..; The roots were immersed in carbol-fuchsin staining solution of 10% leading to a red-violet coloration of the chromosomes. The samples were kept in the refrigerator for 72 hours…. Finally for counting: For mitotic index calculation and chromosomal aberration, 30 microscopic fields were examined for each variant. Three replicates were evaluated for each variant and scoring was performed for two 434 roots per replicate (an average of 3500-4500 cells)…..
I think the authors should describe in the Abstract and the Conclusions the compounds identified by the method used. Such is important.
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to review this manuscript, for your comments and suggestions.
Please find the detailed responses in the attached document and the corresponding revisions/corrections highlighted in the re-submitted manuscript.
Best regards!
Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for Authorshorticulturae-2968862-peer-review-v1
Article : Sustainable Characterization of Some Extracts of Origanum Vulgare L. and Biosafety Evaluation Using Allium Cepa Assay
The work report the potential cytotoxic and genotoxic effects of aqueous and hydroalcoholic extracts of Origanum vulgare L. using the Allium cepa assay.
1- Introduction
1.1- The introduction should be improved, additional references should be incorporated on the uses of these extracts in traditional medicine, justifying the need to develop this study.
1.2- The cytotoxic and genotoxic effects of Origanum vulgare L (extracts and essential oils) as well as the Allium cepa assay have been reported by numerous authors, this gives a lack of novelty to the manuscript.
1.3- A paragraph explaining or highlighting the difference between these results and the numerous trials reported should be included.
2- Table 1. Physico-chemical parameters, total phenolic content and color intensity of oregano aqueous extracts (I01; D02) and hydroethanolic extracts (E03; E04).
It is necessary to apply a simple statistical treatment to these data, to analyze significant differences between the different extraction methods in the different parameters evaluated.
3- Section 1.2. Phytochemical screening of Origanum vulgare L extracts
This section is very preliminary, and is also not very new considering that this plant species has been studied in depth and through solid characterization methods.
Parra, C., Muñoz, P., Bustos, L., Parra, F., Simirgiotis, M. J., & Escobar, H. (2021). UHPLC-DAD characterization of Origanum vulgare L. from Atacama desert Andean region and antioxidant, antibacterial and enzyme inhibition activities. Molecules, 26(7), 2100.
4- A discussion section should be included in the manuscript.
The manuscript in its current state is not appropriate for the Journal.
Lack of novelty and too preliminary
Comments on the Quality of English Language
Minor revision
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to review this manuscript, for your comments and suggestions.
Please find the detailed responses in the attached document and the corresponding revisions/corrections highlighted in the re-submitted manuscript.
Best regards!
Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report
Comments and Suggestions for AuthorsTables 1 and 3:
I am sorry but I do not understand when the authors wrote: Values are significantly different according to one-way ANOVA (*p < 0.05; **p < 0.001). Are they statistically different in relation to what? I deduce that the comparison must be between the averages that are on the same line in the table. But then I do not understand the comparison that is being made. Is it among them? For example, are 0.36 and 0.34 statistically different for p < 0.001?
It is important to enlighten this. The same for Table 3.
I apologize for confusing the authors with two questions, but the main one I had was regarding 420 (yellow), 520 (red), and 620 (blue) nm. However, the new sentence's explanation for the reading range of 190–440 nm was done quite nicely.
Regarding the wavelengths: Calculated as sum of the absorbance at 420 (yellow), 520 (red) and 620 (blue) nm (values registered for each absorbance not shown).
My question was about these readings because the references cited in the article (50, 51) reported reading for wines and different beverages and after reading these references, the samples were red, I am not sure if this is appropriate to use this method for doing the colour of oregano extracts. In the reviewed version there are references 53 and 54 and in both the samples were red or blue… Regarding reference 55, I did not see any reference to the 420 (yellow), 520 (red) and 620 (blue) nm
Concerning the Figure 10, I would remove it, it is not necessary to add it.
Concerning the: a fast and easy-to-handle method, with reliable results, minimal cost, and environmentally friendly.
The authors wrote in the reply: If necessary, we can add to our manuscript the 2 mentioned articles.
It is not necessary, the justification was well done.
Regarding Figure 1, I do agree with the justification made by the authors.
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to read the revised manuscript, for all your comments and suggestions.
Please find the detailed responses in the below attached file.
Best regards!
Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsThe authors have incorporated the suggestions made and justified the questions. The manuscript should be accepted in its current state
Comments on the Quality of English Language
Minor editing of English language required
Author Response
Dear Madame/Sir,
Thank you very much for taking the time to read the revised manuscript, for all your comments and suggestions.
We are glad that we managed to answer your questions clearly.
The manuscript was carefully read and some changes related to the English language were made as mentioned in the attached file.
Best regards!
Author Response File: Author Response.pdf
