Essential Oil of Citrus aurantium L. Leaves: Composition, Antioxidant Activity, Elastase and Collagenase Inhibition
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Round 1
Reviewer 1 Report
The manuscript entitled "Essential Oil of Citrus aurantium L. Leaves: Composition, Antioxidant activity, Elastase and Collagenase inhibition" is a short but comprehensive one. Address a restricted topic. This manuscript may be published after a short revision:
In my opinion, too many unjustified bibliographic sources are used in the introduction. For a statement we have 12 bibliographic sources. It is an exaggeration, especially since these sources are not from the last 10 years.
Table 1 lacks statistical analysis
The conclusion is too short, but the main idea is presented.
The list of references must be updated, on such a topic I recommend that at least 50% of the sources be from the last 10 years.
Author Response
Dear Reviewer,
Thank you for your valuable remarks.
the answers to you comments are below.
Best regards,
In my opinion, too many unjustified bibliographic sources are used in the introduction. For a statement we have 12 bibliographic sources. It is an exaggeration, especially since these sources are not from the last 10 years.
Table 1 lacks statistical analysis
Answer : Standard deviations were added.
The conclusion is too short, but the main idea is presented.
The list of references must be updated, on such a topic I recommend that at least 50% of the sources be from the last 10 years.
Answer : This was modified accordingly. In the present version there are 11/59 (less than 19%) references that are not from the last 10 years.
Reviewer 2 Report
The manuscript entitled “Essential Oil of Citrus aurantium L. Leaves: Composition, Anti-2 oxidant activity, Elastase and Collagenase inhibition.” is well-written and presented interesting data about the essential oil and biological activities of Citrus aurantium. This article could be accepted for publication in Agronomy after a Minor Revision
General comments
1. When it is written Citrus species, make the Citrus in italic type
2. Make the name of the compounds both in the text and the Table in lowercase letters.
3. The Experimental index of each compound which you found should be written in Table 1.
Author Response
Dear Reviewer,
Thank you for your judicious remarks.
The answers to the recommendations are below.
Best regards,
General comments
- When it is written Citrus species, make the Citrus in italic type
Answer : this was corrected
- Make the name of the compounds both in the text and the Table in lowercase letters.
Answer : this was modified accordingly.
- The Experimental index of each compound which you found should be written in Table 1.
Answer : This was added as requested
Reviewer 3 Report
In this article, Oulebsir et al. described the phytochemical analysis of Citrus aurantium L. leaves essential oil by GC-MS technique. Moreover, they analyzed the EO phenol content and its antioxidant activity founding a level lower than EO extracted from peels and flowers. To note, the EO from citrus leaves showed an interesting elastase and collagenase inhibition.
English language and style are fine/minor spell check required:
For example in line 46 put a space between “invaluable and source” words; line 107 delate the period after the [35] reference and so on.
Abstract:
Line 18: “traditional medicine” have to be changed or explained in the introduction. In particular, the authors have to indicate for which diseases and in what route of administration is used.
Line 26: spelling “GAE”
Introduction:
Lines 75-78: add a reference
Line 47: add an appropriate reference to describe the pharmaceutical use
Rephrase lines 58-59
Line 64: it would be interesting analyze the anti-elastase and anti-collagenase activity also for the EO extracted from citrus peels and flowers
Materials and Methods
Line 77: the authors have to explained the method to ground the leaves
Line 84: the formula reported weight of dry sample but the authors did not explained the drying process. When and how it was performed?
Lines 86-87: the authors have to explain because they dried the essential oil
Lines 100-104: why did not the authors use the standard to identify the volatile molecules?
Rephrase lines 116-117
Line 130: spelling ABTS
Lines 132-135: it seems that the ABTS-stock solution is PBS; clarify it
2.7 paragraph: the authors have to describe in detail also the collagenase and elastase assays
2.8 paragraph: The statistical analysis have to be performed for all the experiments and not only for collagenase and elastase tests
Results and discussion
Line 180: the authors reported 45 constituents, but in table they reported 43 elements; clarify
Rephrase lines 190-191
Rephrase lines 196-197: the authors reported “human health” but if the experiments are conducted in vitro so they have to change this sentence
Lines 201-202: add a reference
Lines 206-207: the antioxidant activity obtained in this paper cannot be compared with the [28] reference because the source and extraction methods of EO are different
Rephrase lines 208-219
Table2: replace the table with a figure. Explain in legend the “a, b and c” letters
Lines 245-253: add appropriate references
Lines 264-267: the authors described the use of EO to treat different diseases, it is necessary to describe also the route of administration and the dosage.
Author Response
Dear Reviewer,
Thank you for your judicious remarks.
The answers to the recommendations are below.
Best regards,
For example in line 46 put a space between “invaluable and source” words; line 107 delate the period after the [35] reference and so on.
Answer : Thank you for you remark. This was modiefied in whole manuscript
Abstract:
Line 18: “traditional medicine” have to be changed or explained in the introduction. In particular, the authors have to indicate for which diseases and in what route of administration Answer : this sentence was deleted in order to limit confusion
Line 26: spelling “GAE”
Answer : this was added
Introduction:
Lines 75-78: add a reference
These lines are in M&M. But in you mean L35 to 38, to references were added.
Line 47: add an appropriate reference to describe the pharmaceutical use
Answer : A right reference was added
Rephrase lines 58-59
Line 64: it would be interesting analyze the anti-elastase and anti-collagenase activity also for the EO extracted from citrus peels and flowers
Answer : This sentence was modified
Materials and Methods
Line 77: the authors have to explained the method to ground the leaves
Line 84: the formula reported weight of dry sample but the authors did not explained the drying process. When and how it was performed?
Answer : Thank you for your judicious remark. Indeed this information have been missed in the former version of the manuscript. The drying and grinding methods hzve been added in M&M section.
Lines 86-87: the authors have to explain because they dried the essential oil.
Answer : This step was added in order to improve EO conservation.
Lines 100-104: why did not the authors use the standard to identify the volatile molecules?
Rephrase lines 116-117
Answer : this sentence was clarified
Line 130: spelling ABTS
Answer : this was added
Lines 132-135: it seems that the ABTS-stock solution is PBS; clarify it
Answer : a text was inadvertently added. This text has been removed. The preparation method is now well described.
2.7 paragraph: the authors have to describe in detail also the collagenase and elastase assays
2.8 paragraph: The statistical analysis have to be performed for all the experiments and not only for collagenase and elastase tests
Answer : this was modified accordingly
Results and discussion
Line 180: the authors reported 45 constituents, but in table they reported 43 elements; clarify
Answer : this was corrected. Indeed 43 compounds were identified as presented in table 1.
Rephrase lines 190-191
Answer : this was modified accordingly
Rephrase lines 196-197: the authors reported “human health” but if the experiments are conducted in vitro so they have to change this sentence
Answer : this was modified accordingly
Lines 201-202: add a reference
Answer : two refernces were added
Lines 206-207: the antioxidant activity obtained in this paper cannot be compared with the [28] reference because the source and extraction methods of EO are different
Answer : Thank you for this valuable remark. We have underlined this point in the text. This point is not relly a comparison but information given to readers who are interested to have wide reports for more details.
Rephrase lines 208-219
Answer : this was rephrased as requested
Table2: replace the table with a figure. Explain in legend the “a, b and c” letters
Answer : this was modified accordingly
Lines 245-253: add appropriate references
Answer : references were added as requested
Lines 264-267: the authors described the use of EO to treat different diseases, it is necessary to describe also the route of administration and the dosage.
Answer : this information was added as requested
Reviewer 4 Report
the manuscript deals with the activities of EO extracted from leaves of Citrus aurantium (sour orange) . Only Eos were analyzed, not the true leaves. some activities as anti- elastase and collagenase are detected in Eos.
some parts need an improvements:
1- plant materials: how many plants were used for the collection of he material? do you know the variety of the sour orange?
2-- a comment of the author is required about the use of plastic bags to transport the plant materials. Plastic bags produce big amount of phtalate. 3- add in the supplementary material a chromatogram of the GC-MS in order to verify the absence of any phtalate.
4- table 1: an improvement with more details of the analysis is required in the legend. It is alo recommended to add at each compound the group (id Mh, MO, Sh, SO , non terpenes and other) and also a Linear retention time of a reference (ADAMS or NIST).
5- add at least 3measures and add the average and SD of each single compound.
6- statistical analysis: three replicates; are these biological samples of or technical replicates?
many typos in the text: as
line 60: antifongal instead of anti fungal
line 106: Folin instead of Follin.
line 107 And.
line 109 2ml instead of 2 ml and other similar.
line 201 werechosen
line 212 antoixidant
line 260 fromleaves
line 275 anti-elsatase
Author Response
Dear Reviewer,
Thank you for your valuable remarks.
The answers to the recommendations are below.
Best regards,
the manuscript deals with the activities of EO extracted from leaves of Citrus aurantium (sour orange) . Only Eos were analyzed, not the true leaves. some activities as anti- elastase and collagenase are detected in Eos.
some parts need an improvements:
1- plant materials: how many plants were used for the collection of he material? do you know the variety of the sour orange
Answer : the studied variety is Amara. This was added in M&M
2-- a comment of the author is required about the use of plastic bags to transport the plant materials. Plastic bags produce big amount of phtalate.
Answer : thank you for this interesting remark. The transport time from the samples harvest site to the laboratory is 10 minutes. Considering, the time of harvest and the time of transport we are at 1/2 hour. the samples were rinsed with water before drying. does this delay induce a high concentration of phthalates, it is not conclusive. The chromatogram does not show the presence of phtalates.
3- add in the supplementary material a chromatogram of the GC-MS in order to verify the absence of any phtalate.
Answer : A chromatogram was added as a supplementar material as requested
4- table 1: an improvement with more details of the analysis is required in the legend. It is alo recommended to add at each compound the group (id Mh, MO, Sh, SO , non terpenes and other) and also a Linear retention time of a reference (ADAMS or NIST).
Answer: Table 1 was reformatted and additional details were added as recommended.
5- add at least 3measures and add the average and SD of each single compound.
Answer: this was added.
6- statistical analysis: three replicates; are these biological samples of or technical replicates?
Answer: for the essential oil extraction, three biological samples. For antioxidant, antielastasse and anticolagenase three technical replication were performed for each sample of essential oil. As technical replicates were quite similar, statistical analyses were done on only of biological samples.
many typos in the text: as
line 60: antifongal instead of anti fungal
Answer : this was corrected
line 106: Folin instead of Follin.
Answer : this was modified
line 107 And.
Answer : this was modified accordingly
line 109 2ml instead of 2 ml and other similar.
Answer : this was modified in whole manuscript
line 201 werechosen
Answer : this was modified
line 212 antoixidant
Answer : this was corrected
line 260 fromleaves
Answer : this was changed
line 275 anti-elsatase
Answer : this was modified
Round 2
Reviewer 3 Report
The authors exhaustively replied to the requests
