Review Reports

Round 1

Reviewer 1 Report

  The MS “Inter-varietal variation in phenolic profile, sugar contents, antioxidant, antiproliferative and antibacterial activities of selected Brassica species” has some interests in measuring the nutritional variation between five varieties of Brassica genus, and useful for the local communities. Whereas I have some concerns as follows.

1. Figure 1 and figure 2 were not required to present in the main text.
2. Why one curve is ascorbic acid and the other five are varieties, it is misleading. And there is no variance in it. How many times are repeated about these data.
3. Some references are old, please replace those with news in recent years
4. I can not find the discussion part in the text.

Author Response

Reviewer 1:

Comment: Figure 1 and figure 2 were not required to present in the main text.

Response: Both figures have been deleted as suggested by reviewer 1.

 

 

Comment: Why one curve is ascorbic acid and the other five are varieties, it is misleading. And there is no variance in it. How many times are repeated about these data.

 

Response: In reducing potential assay, the extracts of all the five species were compared with the standard antioxidant (Ascorbic acid) and results are presented in Figure 2. The experiment was performed in triplicate and the data is presented as mean.

 

 

Comment: Some references are old, please replace those with news in recent years

 

Response: References are now updated as suggested by reviewer 1.

 

 

Comment: I cannot find the discussion part in the text.

 

Response: The section is revised, and the discussion is added along with results.

Reviewer 2 Report

In the study, five Brassica samples including cauliflower, broccoli, red cabbage, white cabbage and Chinese cabbage were selected to dertermine the TPC, TFC, phenolic profile, sugar composition, antioxidant activity, antiproliferative activity and antibacterial activity to compare the Inter-varietal variation.

 

However, some parts of the manuscript make me very confused.

 

1. The introduction section was illogical and repetitive.

 

2. There were so many basic error occured throughout the manuscript like mistakes in italics of latin names (Line 51-52, ...), misusing of brackets (Line25, 67, 74,...), missing of commas (Line 21,42,...), and so on.

 

3. Was there MDA-MB231 cells or human A549 lung cancer cells used in 2.10.1?

 

What was the concentration of plant extract used to treat the cells in MTT assay? 40 µg/mL or “40 µg/mL of plant extract, from which 0.5 µL was picked up and diluted in 100 µL cells”?

 

Did the data in Tab. 5 support the conclusion “A pronounced antiproliferative efficacy was showed by cauliflower, Broccoli, red cabbage, white cabbage and Chinese cabbage at (40 µg/mL extract concentration) against the human A549 cancer cell line.(Line 325-327)”?

 

Was it possible that the concentration of plant extract can inhibit not only A549 lung cancer cells but also normal cells?

 

4. The data in Tab. 1 showed that TPC of red cabbage was about 33 mg/g dry material, and the data in Tab. 2 showed that total amount of phenolic componds of red cabbage was about 3.99 mg/g, only 12.1% of TPC.

 

Was it appropriate to state “Catechine was the major flavonoids while gallic acid, p-coumaric acid, chlorogenic acid and benzoic acid were the major phenolic acids found in these Brassica species.”?

 

5. Was the sugar 98.99 g of 100g of dry weight of plant material in red cabbage in Tab. 3?

 

If yes, how can the extract exert the antibacterial activity? 

Author Response

Reviewer 2:

Comment: The introduction section was illogical and repetitive.

 Response: The introduction section is revised and efforts were made to make it logical and comprehensive. The repetition is also managed. The changes are marked with colored font.

 

Comment: There were so many basic error occured throughout the manuscript like mistakes in italics of latin names (Line 51-52, ...), misusing of brackets (Line25, 67, 74,...), missing of commas (Line 21,42,...), and so on.

Response: The manuscript is revised carefully, and all the spelling and grammatical mistakes are now removed from modified version of manuscript.

 

Comment: Was there MDA-MB231 cells or human A549 lung cancer cells used in 2.10.1?

 Response: It is Human A549 lung cancer cells.

 

Comment: What was the concentration of plant extract used to treat the cells in MTT assay? 40 µg/mL or “40 µg/mL of plant extract, from which 0.5 µL was picked up and diluted in 100 µL cells”?

 Response: Stock solution of 40 mg/mL was prepared and from which 0.5 µL was picked. The section is revised to make it clear in revised version of manuscript.

 

Comment: Did the data in Tab. 5 support the conclusion “A pronounced antiproliferative efficacy was showed by cauliflower, Broccoli, red cabbage, white cabbage and Chinese cabbage at (40 µg/mL extract concentration) against the human A549 cancer cell line.(Line 325-327)”?

 Response: The conclusion section is revised as per suggestion.

 

Comment: Was it possible that the concentration of plant extract can inhibit not only A549 lung cancer cells but also normal cells?

 Response: It may be possible, but in the present study, only A549 lung cancer cell line is used. It is good suggestion, and it will be considered in the future work where our team is working on deep down mechanism of anticancer activity.

Comment: The data in Tab. 1 showed that TPC of red cabbage was about 33 mg/g dry material, and the data in Tab. 2 showed that total amount of phenolic compounds of red cabbage was about 3.99 mg/g, only 12.1% of TPC.

 Response: Data in table 1 showed the amount of total phenolic acids present in the plant extracts, as measure spectrophotometrically. Well reported Folin reagent method is used to quantify the total phenolic compounds.

However, the extracts were further subjected to HPLC analysis to quantify the concentration of specific phenolic acids present in each extract. This method did not show the amount of total phenolic acids but very few phenolid acids (11) were separated and detected on High Performance Liquid Chromatography and reported.  The sentence is revised to address the confusion.

 

Comment: Was it appropriate to state “Catechine was the major flavonoids while gallic acid, p-coumaric acid, chlorogenic acid and benzoic acid were the major phenolic acids found in these Brassica species.”?

 Response:

We appreciate reviewer for highlighting these important points related to major phytoconstituents. In revised version of manuscript, it has been mentioned that Catechine and quercetin were the major flavonoids while gallic acid, p-coumaric acid, chlorogenic acid and benzoic acid were the major phenolic acids found in different Brassica species.

Comment: Was the sugar 98.99 g of 100g of dry weight of plant material in red cabbage in Tab. 3?

 If yes, how can the extract exert the antibacterial activity? 

Response: Author admits this mistake and was reported by mistake. The sugar composition was analyzed on HPLC and the results are reported using normalization technique. The results showed the composition of specific sugar (e.g. fructose) in the total detected sugar components. The section and table is revised and results are made clear in revised version of manuscript.

Reviewer 3 Report

The manuscript entitled “Inter-varietal variation in phenolic profile, sugar contents, antioxidant, antiproliferative and antibacterial activities of selected Brassica species” aims to evaluate the variation in nutritional profile, antioxidant activity and free radical scavenging capacity in selected species of the Brassica genus. The manuscript is written well overall. Still, I have some major concerns. The polyphenol and antioxidant profile of Brassica species are well documented in scientific literature, so this manuscript has not much novelty. The authors should emphasize more what is new in their research. I see the strength of this paper is a thorough comparison of these species profiles.

Comments:

-The names and the surnames of some authors are written together.

-Functional food is not a new term.

-“Functional foods save the mankind from the oxidative stress and several diseases like cancer, hemorrhoids, heart diseases, acid reflux, diabetes and stomach ulcer” – the statement is too strong.

-The authors should provide the HPLC profile of the plant extracts.

-Also, some figures would be good to have some kind of evidence for antibacterial and antiproliferative activity.

Author Response

Reviewer 3:

Comment 1:  The names and the surnames of some authors are written together.

Response: Corrections have been made

 

Comment 2: Functional food is not a new term.

Response: Introduction section is revised as per suggestion of other reviewers as well.

 

Comment: “Functional foods save the mankind from the oxidative stress and several diseases like cancer, hemorrhoids, heart diseases, acid reflux, diabetes and stomach ulcer” – the statement is too strong.

Response: Introduction section is revised as per suggestion of other reviewers as well.

Comment: The authors should provide the HPLC profile of the plant extracts.

Response: The Typical HPLC profile showing the separation of phenolic acids and flavonoids is added in the manuscript as Figure 1.

Comment: Also, some figures would be good to have some kind of evidence for antibacterial and antiproliferative activity.

Response: The manuscript now contained 5 tables and 2 figures. The antibacterial and antiproliferative data is presented in tables.

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Please check the manuscript carefully to eliminate undeserved mistakes. 

Author Response

Reviewer 2

Comment: Please check the manuscript carefully to eliminate undeserved mistakes. 

Response: The manuscript has been revised carefully and more improvements have been made and highlighted with blue font

Reviewer 3 Report

The authors made some improvements, but they still need to address the novelty issue, the most important comment. Also, the figure with antibacterial activity results needs to be added.

 

Author Response

Reviewer 3

Comment: The authors made some improvements, but they still need to address the novelty issue, the most important comment.

Response:

Although some work has been conducted separately on different varieties of Brassica species but the present study was aimed to investigate inter-varietal variation in the total phenolic and total flavonoid contents, antioxidant and free radical scavenging potentials, anti-proliferative, antibacterial activities of some cauliflower, red cabbage, Chinese cabbage, white cabbage and broccoli. No such data is present in the single paper. Moreover, the identification and quantification of phenolic acids and flavonoids of all selected species varieties was carried out in a single run by reverse phase High performance liquid chromatography (RP-HPLC). Therefore in the present manuscript, the sugar and phenolic profiles, antioxidant, antibacterial and anti-proliferative activities of five locally grown Brassica species, have been presented1st time.  

 

Comment: The figure with antibacterial activity results needs to be added.

Response: Typical figures have been added in the revised manuscript as Figure 3.

Author Response File: Author Response.docx