Optimization of the Multienzyme-Assisted Extraction Procedure of Bioactive Compounds Extracts from Common Buckwheat (Fagopyrum esculentum M.) and Evaluation of Obtained Extracts

Round 1

Reviewer 1 Report

The authors did a good work from an experimental point of view and I recommend the article for publication after some minor revisions.

More specific:

L47: Replace the ‘’quercetin-3-rutinosid’’ with ‘’quercetin-3-rutinoside’’.

L53: Add the ref. Zapata-Morales et al. [18]. No dot between et al.

L195: Provide some product details from the manufacturer, such as nutritional facts, organic or not, product name, etc.

Author Response

Thank you for Your comments

Author Response File: Author Response.docx

Reviewer 2 Report

I would suggest that the authors correct the title of the manuscript. Since the meaning of it reads as "buckwheat optimization". The same expression is found in the abstract. It would be more logical to replace it with "Optimization of the extraction procedure using ..."

 

The separate parts of the introduction are not logically connected. In addition, the introduction contains too general information.

 

Line 55 Recent studies also determined the technological properties of rutin and soy protein interaction due to stabilized foam formation [19].

In this case, a link to several sources is required.

 

The introduction does not quite clearly lead readers to the stated goal of the study. In addition, the introduction lacks the hypothesis of the study. Would recommend that authors add this paragraph to the introduction.

 

"Non-starch polysaccharides enzyme mixture"

Are there any known components of such a mixture, in addition to the fact that it contains cellulases and xylanases. This would allow the authors to better discuss obtained results.

 

There is no regression equation. When presenting results as response surfaces, the regression equation must be presented.

 

Despite the fact that the chapter is stated as “results and discussion”, there is actually no discussion.

In particular, as one of the elements that should be reflected in the discussion, for example a comparison of the presented method with other methods of extraction of phenolic compounds that are used in the food industry (for example, ultrasonic micellar extraction, combined extraction approaches etc).

 

Figure 1

Coarse figure caption. The figure at least lacks the designation of the main parameters, which are coded with letters in the figure. Not very good resolution of figures, small symbols of values on the axes.

The maximum yield (Figure 1, a) can be observed at the minimum temperature (60) and the maximum processing time (24 hours). However, this point is not the surface apex. This may indicate that the parameters were chosen incorrectly. For example, a lower extraction temperature should have been used and processed for a longer time.

 

Figure 2

Rutin concentration scale. Leave only integer values.

The character of the distribution of letters is not clear when assessing the reliability of differences.

The figure captions are not informative.

The boxes around the histograms look superfluous.

You can use one legend for all three graphs.

There is a certain confusion in the abscissa axis notation, for example, what is the difference between control and 0.

On the time axis, it would be possible to leave only 3 significant points.

 

Figure 3.

Near the corners there are images that do not carry a semantic load, they look like artifacts. If they are still meaningful, it is necessary to increase the resolution, if not, remove them from the photographs.

There is no scale bar.

The enumeration of the experimental variants themselves (such as “Control sample solid fraction”) can be transferred to the figure caption so as not to overload the graphic image with additional information.

 

The division into groups (I, II, III) is not entirely clear. What is the difference between them.

Author Response

Thank you for Your comments. Please see the attachment

Author Response File: Author Response.docx

Reviewer 3 Report

Dear Authors, 

I recommend your manuscript to be published at present form

Author Response

Thank you 

Reviewer 4 Report

The work is interesting but some corrections are needed

• sentence in lines 47-48 - is unclear.
• Figure 2 is opaque and unclear. the primary y-axis should also be an integer, also the legend below or below the graph will give a clearer view of the results, and complete ambiguity is introduced by the solid line / time. The time cannot be shown by a line if it is not continuous, and it is not because the control time and NPS is 25 h and these two times cannot be connected by a line! It is also unclear why the x-axis is NPS, when NSP is mentioned in the materials ?!
• Figure 3. the sign "°ˇis mising on the first control sample. The sign for angle must stand next to the number

Sincerely 

Author Response

Thank you for Your comments. Please see the attachment

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Title «evaluation of physicochemical characteristics ...

In my opinion, "of obtained extracts" should be added.

The introduction still does not spell out the hypothesis of the study, although such recommendations were made in the first round review.

Please show the results of statistical analysis (Tukey's multiple comparison test.). Please report the p-value obtained by ANOVA for each measured parameter in Supplementary.

Whether the adequacy of the developed model has been assessed? Is there a functional relationship between the experimental factors and the response variable?

Have the coefficients of determination (R ^ 2), adjusted R2 (R ^ 2 (adj).)) and predicted R ^ 2 (R2 (predicted)) calculated to validate model fit. These data should be presented in the manuscript.

I suppose that the equations in the text of the manuscript are formatted in a different way (not like figures). Please check the template.

Remove frames around graphs

As before, the part related to the discussion of the results obtained is very weak.

1,2,3 are Arabic numerals, not Roman (as stated in the review response). Write down in the text of the sub-chapter the conditions for obtaining extracts before the analysis, since they are mentioned only in the table.

Author Response

Thank you for your comments, please see the attachment below

Author Response File: Author Response.docx

Reviewer 4 Report

My previous comment related to previous figure 2, not figure 3 was: The "time" cannot be shown by a line if it is not continuous, and it is not because the control time and NPS is 25 h and these two times cannot be connected by a line! 

- but it is still presented in a for of a line (Fig 3). 

it may be an oversight, but the reader of your paper will think it is correct and the error will be passed on.

Still are present markings on the x-axes that are NSP and NPS?

 

Please correct it.

 

Author Response

Thank you for your comments, please see the attachment 

Author Response File: Author Response.docx

Round 3

Reviewer 2 Report

Figure 2
I still do not understand the nature of letters distribution which indicate the significant difference between samples. There should be some regularity. For example, the letter "a" can denote the variant or variants with the highest values of the concentration of rutin.

Figure 3
or 4 (in the text under the figure, it is indicated this way). There is a lack of a full-fledged legend for this drawing. In addition, it is extremely untidy. I took it upon myself to prepare an approximate version of such a drawing (see the attached file). I would like to draw the attention of the authors to some details.
-The distance between the individual figures must be the same.
- The frame with the designation of the angle, in all figures, should be in the same place and of the same size. It is enough to indicate only degrees there.
- It is necessary to provide a scale bar in the figure.
- Make the lines for marking the angle clear.
- Assign a corresponding indicator to each figure, which corresponds to the sample variant

Comments for author File: Comments.pdf

Author Response

Thank you for your comments, please see the attachment

Author Response File: Author Response.docx

Reviewer 4 Report

Your engagement in corrections is very commendable, resulting in a very meaningful presentation of your valuable study. Well done!

Author Response

Thank you!

Round 4

Reviewer 2 Report

The authors have done a great job to improve the quality of the manuscript.
One last note: Figure 3, draw a segment that characterizes the scale bar, and it is better that this value is a multiple of 5 (for example, 10 mm). Otherwise, it is not clear what 6 mm means: this white square or the size of a drop on the surface.