Use of Unmalted and Malted Buckwheat in Brewing

Round 1

Reviewer 1 Report

Reviewer’s Comments:

The manuscript “Use of unmalted and malted buckwheat in brewing” is very interesting work. This paper investigates the new assortments are created using different combinations of ingredients or changes in the brewing process. In recent years, research studies on the use of buckwheat in brewing, especially gluten-free beer, have intensified considering the increase in the number of people suffering from celiac disease. This paper proposed the use of malted and unmalted buckwheat in brewing, the results obtained being compared with beer made from 100% malted barley. In order to improve the production yield, the technological process of mashing was optimized by adding commercial enzymes. Buckwheat use was facile, the technological process was similar to barley and the finished product more or less resembled typical beer in many ways, which makes buckwheat suitable for habitual beer consumers who seek a traditional and well-established beer properties. Beer - finished product presented physico-chemical characteristics close to those of beer obtained from 100% barley malt.  However, the following issues should be carefully treated before publication.

1. In abstract, the author should add more scientific findings.

2. Keywords: the synthesized system is missing in the keywords. So, modify the keywords.

3. In the introduction part, the introduction part is not well organized and cited references should cite the recently published articles such as 10.1016/j.inoche.2022.109449 and 10.3390/bioengineering9100486

4. Introduction part is not impressive and systematic. In the introduction part, the authors should elaborate the scientific issues in the malted buckwheat research.

5. Evaluation of the quality of buckwheat and buckwheat malt …, The author should provide reason about this statement “The thousand corn weight is much lower, by 30.78%, in buckwheat compared to barley, due to the size of the grains”.

6. The authors should explain regarding the recent literature why “Energy and germination ability recorded higher values for buckwheat than for barley, so, in conclusion, the analyzed buckwheat can be subjected to the malting process”.

7. Experimental brewing with buckwheat malt and buckwheat. The author should explain the latest literature “The time from reaching the temperature of 72°C until the decomposition of the starch (negative iodine test) is called saccharification time (rate). This indicator is a general measure of starch breakdown, dependent on the activity of amylolytic enzymes in the mash”.

8. The author should provide reason about this statement, “The total nitrogen content of the beer will depend on both the raw materials used during brewing and the parameters of the brewing process”.

9. Comparison of the present results with other similar findings in the literature should be discussed in more detail. This is necessary in order to place this work together with other work in the field and to give more credibility to the present results.

10. The conclusion part is very week. Improve by adding the results of your studies.

Author Response

Response to Reviewer 1

Dear Referee,  

We would like to thank the referee for the close reading and for the proper suggestions.

We hope that we provide all the answers to the reviewer’s comments.

Thank you very much for the recommendations to publish our paper entitled “Use of unmalted and malted buckwheat in brewing”.

The present version of the paper has been revised according to the reviewer’s suggestions.       

We uploaded the corrected version of the article for which we used the red colour for the addition text.

Kindly take notice of these changes made to the manuscript's text. The red colour indicates the answer to your suggestions, while the green and blue colours indicate the responses to the recommendations of the other three reviewers. We regret not include the responses to your suggestions in this letter. The paper (V2) has been attached with all the revisions made per the suggestions of the four reviewers.

Point 1: In abstract, the author should add more scientific findings.

Response 1: First of all, we would like to thank the referee for the close reading and for all the given comments suitable for improving the manuscript.

We made the changes according to the referee suggestions.

Point 2: Keywords: the synthesized system is missing in the keywords. So, modify the keywords.

Response 2: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions.

Point 3: In the introduction part, the introduction part is not well organized and cited references should cite the recently published articles such as 10.1016/j.inoche.2022.109449 and 10.3390/bioengineering9100486

Response 3: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions.

Point 4: Introduction part is not impressive and systematic. In the introduction part, the authors should elaborate the scientific issues in the malted buckwheat research.

Response 4: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions. We added and other scientific reports about unmalted and malted buckwheat beer.

Point 5: Evaluation of the quality of buckwheat and buckwheat malt …, The author should provide reason about this statement “The thousand corn weight is much lower, by 30.78%, in buckwheat compared to barley, due to the size of the grains”.

Response 5: We would like to thank to the referee for her/his remarks. We corrected it by excluding this characteristic from the text.

Point 6: The authors should explain regarding the recent literature why “Energy and germination ability recorded higher values for buckwheat than for barley, so, in conclusion, the analyzed buckwheat can be subjected to the malting process”.

Response 6: We would like to thank to the referee for her/his remarks. We corrected it by excluding these characteristics from the text.

Point 7: Experimental brewing with buckwheat malt and buckwheat. The author should explain the latest literature “The time from reaching the temperature of 72°C until the decomposition of the starch (negative iodine test) is called saccharification time (rate). This indicator is a general measure of starch breakdown, dependent on the activity of amylolytic enzymes in the mash”.

Response 7: We would like to thank to the referee for her/his remarks. We added the explanation in the article.

Point 8: The author should provide reason about this statement, “The total nitrogen content of the beer will depend on both the raw materials used during brewing and the parameters of the brewing process”.

Response 8: We would like to thank to the referee for her/his remarks. A mistake was made here, it is not about the nitrogen content of the beer but the nitrogen content of the wort. We corrected it.

Point 9: Comparison of the present results with other similar findings in the literature should be discussed in more detail. This is necessary in order to place this work together with other work in the field and to give more credibility to the present results.

Response 9: We would like to thank to the referee for her/his remarks. We have also added other bibliographic references.

Point 10: The conclusion part is very weak. Improve by adding the results of your studies.

Response 10: We would like to thank to the referee for her/his remarks. We have improved this part of article.

Author Response File: Author Response.pdf

Reviewer 2 Report

The submitted work is quite interesting for the readership of Applied Sciences.

I suggest minor corrections on English language and shortening of the paper, the length is far too long on comparison with the significance of the paper. 

Author Response

Dear Referee,  

We would like to thank the referee for the close reading and for the proper suggestions.

We hope that we provide all the answers to the reviewer’s comments.

Thank you very much for the recommendations to publish our paper entitled “Use of unmalted and malted buckwheat in brewing”.

The present version of the paper has been revised according to the reviewer’s suggestions.       

We uploaded the corrected version of the article for which we used the red color for the addition text.

 Reviewer 2: I suggest minor corrections on English language and shortening of the paper, the length is far too long on comparison with the significance of the paper. 

 Response 1: First of all, we would like to thank the referee for the close reading and for all the given comments suitable for improving the manuscript.

We made the changes according to the referee suggestions.

Reviewer 3 Report

The aim of the investigation is unclear.

Abstract

13- craft and functional beer have revived

26 – I do not recall any data on the ‘purity’ of taste, nor ‘tingling’

28-29 – this is not in line with your results. The use of more unmalted buckwheat lead to faster filtration (table 6)

24 – “beer – finished product”, you could use “final beer” instead.

24 – Could you be more specific? For example, mention which characteristics are similar, or which

characteristics differ

The aim of the investigation is unclear: ‘this paper proposed the use of malted and unmalted buckwheat’ -> this has been proposed before, what is the scope of this investigation? From this abstract, it cannot be deduced that different proportions/combinations of malted and unmalted buckwheat were being compared. It seems here that you are going to compare 100% malted buckwheat, 100% unmalted buckwheat and 100% barley malt. I think it would be useful to clearly state the scale of the experiments since it appears that brewing with buckwheat on a larger scale 20- 60-500 L is not always obvious.

1. Introduction

36 – citations [2-5] are not relevant as those studies are not focused on the beer consumption

39 – you could use “beer industry” or “brewers” instead of “beer industry processors”

43 – cereals and pseudocereals, as buckwheat is a pseudocereal.

51 – In a brewing context I would use “adjunct” instead of “adjuvant”

55 – I do not think citation [3] mentions this.

58 – Did the literature show that buckwheat has outstanding results for enzymatic activity (you state it otherwise in line 72)? As I recall it, the use of exogenous enzymes is recommended in the literature. Also, in that case, enzymes would not be needed during your experiments. E.g. [33, 19,21, 27, 32, 38,…] and you can also refer to Myncke, E.; Laureys, D.; Baillière, J.; Vanderputten, D.; De Clippeleer, J. Mashing with Unmalted Buckwheat: Influence of Proportion, Grind Size, and Exogenous Enzymes on Small-Scale Lautering Performance. MBAA Technical Quarterly 2022, 59(2), 55-62.

73 – it might be better to refer to more specific research on the enzymatic activity of buckwheat vs barley like Wijngaard et al. and not to a review.

75 – do you mean low levels of fermentable sugar in malt or wort?

77 – also to improve filtration, E.g. Myncke et al.

80 – Adjunct instead of adjuvant

87 – there are already quite some studies on the optimization you could refer to.

89 – there are already quite some studies on the physicochemical and sensory properties you could refer to.

However, mainly for malted buckwheat.

2. Materials and Methods

2.1 Materials

94 – which variety of buckwheat (malt) was used?

100 – Which enzymatic activity does Termamyl Classic have?

2.2 Obtaining beer in laboratory conditions

2.2.1 Mashing

Were the experiments performed in duplicate, triplicate, … ? Please mention this.

108 - Citation [26], use the correct format, as indicated by Analytica EBC (https://brewup.eu/ebcanalytica/about): For example, if you wish to cite Method "4.3.1 - Total Nitrogen of Malt: Kjeldahl Method(IM)" that you have accessed on 11 January 2021, please cites as follows: "EBC Analytica, 4.3.1 - Total Nitrogen of Malt: Kjeldahl Method (IM), 2004, 22 October 2018, accessed on 11 January 2021, https://brewup.eu/ebcanalytica/malt/total-nitrogen-of-malt-kjeldahl-method-im/4.3.1"

109 – Change “laboratory mill” to “laboratory disc mill” and mention the gap between the grinding discs (should be 0.20 mm).

111 - The term "mash beaker” or “mash cup" is perhaps more forthcoming than vats.

112 – Use “mashing bad” instead of “device”, and mashing program instead of flotation program.

117 – Clarify how you determined the duration of saccharification, I assume a simple starch–iodine test.

121 – 126 You might want to explain in the introduction why we need a modified Congress method for buckwheat and unmalted buckwheat. “the sample” did not saccharify, which sample -> 100% buckwheat malt, 100% unmalted buckwheat, … ? Why would the addition of 1 g barley malt lead to saccharification?

The usage of different buckwheat malt/flour proportions or the use of enzymes is not mentioned. Please add this to your materials and methods.

2.2.2 Mash filtration

128 – The term "mash beaker" is perhaps more forthcoming than a flask.

128 – The obtained wort (2.0-2.2 L) is boiled together. When is the content of the mashing cups (and how many beakers) brought together: before or after mash filtration? Please mention this. Also, were the quality characteristics of the wort determined on a mixture of the different mashing beakers, or on each individual wort?

129 - By a pleated filter, don't you rather mean a folded filter? Can you also give some more specifications of this filter (brand, pore size, diameter, ...) and on the fermentation vessels (material, shape, volume, brand,..)?

2.2.3 Wort boiling

How much bitterness (IBU) was aimed at?

133 – The obtained wort (2.0-2.2 L)…

136 – a capacity of 2 L…. (space between 2 and L)

2.3.1 Analysis of raw materials:

150 – “the protein content, and the starch content, were determined for the barley and buckwheat samples”

152 – Citation [25]: Make sure that the analyses, described in lines 149-152 section are not published yet as results. Otherwise, you need to refer to citation [25] and table 1 when describing the used materials (section 2.1). In that case, move Table 1 from "Results and Discussion" to section 2.1, or incorporate it in the introduction.

160 – Citation [26], same comment as before on Analytica EBC.

2.3.2 Analysis of beer:

150 – “O2 content (EBC 9.37.1), and caloric content”

168 – Citation [26], same comment as before.

Was this also done in technical triplicate as the analysis of raw materials and wort? Please mention this.

165- the turbidity Is not measured by EBC 9.43.2, -> 9.43.2 - Specific Gravity of Beer using a Density Meter.

What is the difference between turbidity and turbidity 90/25?

2.3.4 Statistical analysis

181 – it is unclear to me which three values were taken to calculate the mean: in line 146 you state that all laboratory brewing experiments had been repeated three times (biological triplicate) and in line 160 you state that there were three determinations for each sample (technical triplicate). Did you make 3 beers for each variant 1-8 and CS-B4??

181 - Mean +- standard deviation

182 – you did not mention the various levels of (malted) buckwheat used in the mashing experiments. Please mention clearly in section 2.2.1 that the amount of buckwheat (malt) varies and which % are used.

3.1 Evaluation of the quality of buckwheat and buckwheat malt

188 – Rephrase the first sentence.

189 - Something is wrong with the sentence structure: just add ‘unmalted’ to buckwheat or barley

Table 1. Add unmalted to the figure caption

198 – “buckwheat - raw material”, why the “-“?

201 – Are these differences significant? If so, on which level (alfa= ?)?

208 – even higher levels of starch 74% were found by Jeroen Baillière et al (citation [16]); this depends on variety, cultivation region, harvest year, …

205 – “Energy and germination ability” Do you mean germination energy and -capacity?

The data in table 2: Did you determine the quality characteristics of the wort as a mixture of the different mashing beakers, or on each individual wort? Is the mean and standard deviation presented, calculated as the mean of: the different mashing beakers (3x450g) (biological); the at least three times repeated laboratory brewing experiments (3x 2 L scale, line 146), or the three determinations from the same sample in parallel (line 160)? Please clarify this more in the Materials and Methods.

221 – The recommendation for the use of enzymes was also made several times in the literature. E.g. [33,19,21, 27, 32, 38,…] and Myncke, E.; Laureys, D.; Baillière, J.; Vanderputten, D.; De Clippeleer, J. Mashing with Unmalted Buckwheat: Influence of Proportion, Grind Size, and Exogenous Enzymes on Small-Scale Lautering Performance. MBAA Technical Quarterly 2022, 59(2), 55-62. The need for enzymes for filtration and extract issues should be addressed in the introduction.

Overall, you compare buckwheat malt quality to barley malt in table 2, but the used mashing scheme (Congress vs modified Congress) to determine extract, and saccharification time, … differs. You state (216) that the quality indices of buckwheat malt are acceptable, but on what basis do you conclude this, given for instance the long filtration and saccharification times.

234 – the citation to [13] is redundant

3.2 Experimental brewing with buckwheat malt and buckwheat

Table 3, and some more explanation on the buckwheat malt/flour proportion and enzyme addition should be added to section 2.2.1. How are the buckwheat fractions combined? Did you mix the grains in the desired proportions and then mill them? Or did you mill the fractions separately and then mix them together in the desired proportions? If the latter, particle classification will occur in the grist meaning that different proportions of husk and fines will be allocated (systemic error). When are the enzymes added, which enzymes are active in Termanyl classic enzyme preparation and on what did you base your assessment of the range (2-5%) of enzyme addition?

Clearly state the aim of this first experiment. The recipe variants seem quite random, explain why these conditions were selected.

244 – Not only amylolytic enzymes as you are using unmalted buckwheat. Without cell wall degradation and proteolysis during malting, amylolytic enzymes are not able to hydrolyse starch. In the case of unmalted grains, this has to happen during the mashing. The saccharification time will therefore also depend on protease and hemicellulase activities. (read Myncke et al for more info)

247 – The gelatinization temperatures are very important to mention indeed. You might want to add some Tgel for unmalted buckwheat ( citation [16] for instance) and compare these with barley malt. Do you expect any issues when comparing these temperatures with barley malt?

Table 4. – please adjust the significant digits of the mean values to the std. deviation. E.g. time 5±0.58 min -> 5.00±0.58 min

- What does this data represent? Each variant is made 3 times or did you determine each value 3 times on the same variant (unclear from materials and methods)

- Don’t use “beer wort” in the figure caption, it is either beer or wort. What do you mean by “the first experiment”? I see that you refer to the second experiment in table 6, but until now it is unclear that these experiments are some preliminary experiments, and what they aim to show us.

- You could use “unmalted buckwheat” instead of buckwheat flour

251-253 – The sentence structure is not right, rephrase

253 – Use variant 8 instead of last variant

How can it be that, using the same amount of exogenous enzymes, the saccharification time of 100% malted buckwheat exceeds that of 90% malted buckwheat with 10% unmalted buckwheat? You would expect that more unmalted material means less enzymatic activities and longer saccharification times.

255 – “cooling to 20°C”, “450_g”

For the effect on filtration, please read and refer to Myncke et al.

257 – Much more than Beta-glucans alone: other fibres (arabinoxylans, …) but also high molecular weight proteins, starches, …

258 – You often use “adjuvant” what is the difference between adjunct and adjuvant?

260 – You are right to highlight the importance of Beta-glucans for filtration. However, this is mainly the case for unmalted barley. Recent studies [16, Myncke et al.] show very low levels of Beta-glucans for alternative cereals and pseudocereals.

256 – 262 you discuss the effect of composition, … on filtration, but what about your results? What is the effect of proportion unmalted or exogenous enzymes? The filtration times are a lot higher for variant 8, this is again unexpected, as more unmalted material means less enzymatic activities and more long-chained polymers (starches, protein, fibre). I miss some more discussion of the results. The same goes for extract. If you do not discuss your results, section 3.2 is obsolete.

271 – I am not familiar with “must” in a brewing context, do you mean wort?

272 – Do not use “EBC-unit” for colour, just EBC

272 – Pay attention when making the comparison between barley malt and buckwheat (malt). Keep in mind/mention that the wort colour is not only influenced by the raw material but also influenced by the mashing scheme (barley malt wort in Table 2 was produced by Congress and not modified Congress).

The effect of the proportion of buckwheat on pH is also described by Myncke et al.

277 – Some of the effects of lowering the wort pH Some, apply to the mash pH and not the wort pH. (viscosity of mash, enzyme activity, …)

276 – 280 citations missing?

Also, you mention the effect of pH on beer quality, but what may we expect at the resulting pH values for variants 1 – 8, I am missing a discussion of the results

281 – “Phiarais et al. (2010) [38], obtained a beer from 100% buckwheat malt with characteristics like wheat beer in terms of pH, nitrogen content, degree of fermentation and alcohol content. “ This sentence is a bit out of place here, rather something for in the introduction.

I am missing the added value of section 3.2 Experimental brewing with buckwheat malt and buckwheat. What specifically did you want to examine through this experiment? Different parameters are adjusted (% buckwheat, and enzyme) why were these different variants compared? Make it clear that you are doing this to ascertain the correct enzyme dosage, and build the discussion of your results accordingly. Discuss which parameters you will look at to assess enzyme activity (filterability, saccharification, extract, yield). Note that variant 8 reacts very unexpectedly and then conclude that the amount of enzyme dosage does not matter much. Then, also compare this to a variant where no enzymes are administered to show that enzyme use does have an effect.

3.3 Study on the production of beer from buckwheat and buckwheat malt under laboratory conditions

Table 5 on the different variants should be discussed in the materials and methods (like note in table 3)

Why not 100% malted as well? What information are you trying to obtain using these specific recipe variants?

297 – Barley malt is the control sample, but make sure it is clear that another mashing scheme (Congress vs adapted congress) was used compared to buckwheat (malt).

301 – Sentence structure is not correct

301 – Can’t you use statistics to determine if these values are significantly different from the reference CS, instead of “close values”?

303 – Significant difference on which p-level?

306 – Double citation, leave the [18] as you cite to Cela et al. (2022) at the beginning.

Table 6. The results for saccharification time show that (as we would expect) a higher % of malt (more enzymatic activity) results in faster saccharification. This is in contrast to the results in Table 4 where 100% malted buckwheat has the longest sacch. times. The filtration time (unexpectedly) increases when more unmalted buckwheat is added. I miss a more in-depth discussion on the results shown in table 6. What is the effect of the increasing unmalted buckwheat/ buckwheat malt proportion on the different wort parameters, and how to explain these effects? How do these results compare to barley malt?

319 – remove the “in all samples taken in the study”

322 –“ Compared to the wort obtained from barley malt, where the content in the extract increased after boiling to the value of 11°P, all four wort samples had the content in the extract lower, between 11 and 12°P”

Reformulate for sentence structure and comprehensibility.

325 – Similar double citation

326 – 7-8 EBC to 11-12 EBC

Table 8. – for one parameter use the same amount of significant digits for the standard deviation e.g. Real extract: CS: 11.05 ± 0.40 as the standard deviation for the others B1-B4 has 2 sig. digits. Change all “,” to “.”

333 – insignificant on which level? -> it might be opportune to show significant differences from the CS by an * in your tables

337 – “Obtaining different results of this part led to a lower energy value of beer - finished product.” Rephrase

339 – only 1 study not studies; remove the double citations

341 – 344 same, remove double citations

343 adjunct instead of adjuvant

346 which IBU did you aim for when hop was added? First, you state that the IBU decreases by using buckwheat (I would not necessarily conclude this from this data), and then you state that the isohumulone content is similar among the four varieties… However, CS and B1 have both a bitterness around 25 IBU.

348 – What do you mean by “plain” extract -> apparent extract?

We need more discussion of the results from Table 8

-The ethanol content is affected by using buckwheat malt and unmalted buckwheat -> Ok, but was this expected? Why is it affected? Is there also a difference between malted and unmalted buckwheat? .....

- The colour is a result of Maillard, has buckwheat more FAN/reducing sugars compared to barley? What is the effect of malting on colour? Malted buckwheat should add more colour than unmalted buckwheat… is this the case? What is the effect of mashing on the colour, you compare barley with buckwheat but another mashing scheme was used.

Figure 1. Blurred frame lines are visible around some of the graphs. The units are missing in both axes.

Figures general: I am not convinced that these types of figures are best to represent the possible trends. I wonder if 4 data points (4 combinations of unmalted and malted buckwheat) are sufficient to predict the whole range of combinations. A 2D representation with unmalted buckwheat as the x-axis would suffice since the proportion of malted buckwheat is (1 - proportion unmalted buckwheat). From these figures, it is also unclear to me why you do not see/discuss trends e.g. with colour, pH, and oxygen, but you do with the other parameters, for example. Some representations do not make much sense, e.g. CO2: a higher proportion of buckwheat would lead to more CO2, but a higher proportion of unmalted buckwheat would as well.

353 – Which ‘extract’ -> real, apparent, or both?

It is unexpected that the extract content decreases when more malt is used isn't it?

355 – 358 -> Missing citation. It would have been useful to compare the protein and starch content of the used buckwheat (Table1.) and buckwheat malt (not determined?).

358 – “barely losses some amounts of its content which is necessary for embryo development” -> too vaguely

361 – “beer wort” is not a thing, it is either beer or wort. Rephrase for readability (3 times fermentation/fermentable in 1 sentence)

361 – 367 It is unclear what you are trying to say. How can your real extract in Table 1 be higher than the Original extracts in Table 7? I think you use the wrong terminology in table 8 and figure 1. It does not make sense that a higher real extract (= extract remaining in the beer) would lead to higher ethanol content (fewer sugars are fermented, but more ethanol is formed?) However, this goes for the original extract (the real extract BEFORE fermentation) as more sugars are available to produce ethanol.

And what about density? You would not expect density and alcohol content to show a similar trend, as an increase in alcohol content and the reduction in fermentable sugars would lead to a lower density.

368 – “and carbon dioxide level”

Figure 2 Not all axis have units. Blurred frame lines are visible around some of the graphs. It also looks like some data points are missing/coincide in Figures 2 b,c and d?

375 – don’t use “buckwheat malt flour”, until now you distinguish malt and flour. In general, I think it is better to talk about malted and unmalted buckwheat for ease of understanding.

What is the difference between beer turbidity on the one hand, and turbidity S25/90 on the other? You refer to EBC 9.42.1 but this is not correct as stated before. The trend visible in figure 2a is not the same as in figure 2b or 2c but this is not discussed? This makes the following discussion unclear.

377 – Nice hypothesis, and rather easy to verify… you could use the literature (a lot of buckwheat malting research available by Wijngaard) or determine the protein content of the used malted buckwheat yourself to compare it with the unmalted buckwheat (as stated before).

The following discussion is unclear. First, you mention 375-380 higher protein, polyphenolic tannins and soluble nitrogen for unmalted buckwheat, which will lead to haze. Next, you state (381-384) that higher amounts of unmalted buckwheat will prevent haze formation due to a decrease in low-molecular nitrogen and protein.

385 – which “high” level? Be more precise.

384 -389 Isn’t the given explanation contradicting your results? The lowest value in Table 8 is 0.36 for B3 and not the value for B4. Moreover, consider the amount of CO2 formed during fermentation, not only the amount retained. Less fermentable wort will lead to less CO2 formation.

391 – “the various proportions of unmalted and malted buckwheat used”

396 – significantly on which level?

400 – the raw materials used can have a significant effect on the pH…

Figure 3 – be consistent: you use colour (USA) and colour (UK)

418 (and further) Put the descriptive score (like very much, like moderately, ...) in italics or in quotation marks to improve readability

Table 1. Have the various items under "Foreign matter, total, or which %," indented (right-aligned) by the same amount.

420 – remove double citation (also 424). It now seems that your research has also shown that the buckwheat beer shows notes of toasted sunflower seeds, but I don't think this is the case.

428 – Are these results in agreement? Do you have a paler beer with more malted buckwheat? Your results show the opposite when comparing CS to B1-4.

432 – Your data (B1 better than the other B2-B4, or malted buckwheat better than unmalted) is not in agreement with Dezellak et al. as they compared malted buckwheat to quinoa and barley.

460 – Don’t you mean the original extract?

470 – fermentable

479 – analytically determined bitterness instead of physical

480-481 Unclear formulation

484 – Aroma, mouthfeel and bitterness do not determine beer taste. Beer taste is determined by bitterness, salinity, sweetness and sourness. Taste is a part of beer flavour, together with aroma, odour and mouthfeel.

3.4 PCA of the physicochemical and sensory characteristics of beer samples

441-443 Isn't it fairly obvious that the sensory parameters to be surveyed will determine the general acceptability surveyed?

From Figure 5 I would assume B2 is more alike to CS than B1 or B4 (closer to PC2 and PC1)?

3.5 Conclusions

The information 491-494 has not been indicated by your results, or cited/mentioned before and should therefore not be part of your overall conclusions. No new information to the literature has been added to your conclusion.

Author Response

Response to Reviewer 3

Dear Referee,  

We would like to thank the referee for the close reading and for the proper suggestions.

We hope that we provide all the answers to the reviewer’s comments.

Thank you very much for the recommendations to publish our paper entitled “Use of unmalted and malted buckwheat in brewing”.

The present version of the paper has been revised according to the reviewer’s suggestions.       

We uploaded the corrected version of the article for which we used the blue colour for the addition text.

We thought you put a great deal of effort into reviewing the article, and it amply demonstrates your expertise in the research topic.

I gained a lot of knowledge from this evaluation, such as the necessity for more zealous research planning and analysis etc.

We feel that by making the necessary modifications, we have addressed the majority of your requirements. A portion of the errors could also have been brought on by the young PhD candidate's desire to learn more about this brand-new industry being explored in our country: buckwheat beer.

Kindly take notice of these changes made to the manuscript's text. The blue colour indicates the answer to your suggestions, while the red and green colours indicate the responses to the recommendations of the other three reviewers. We regret not include the responses to your suggestions in this letter. The paper (V2) has been attached with all the revisions made per the suggestions of the four reviewers.

Author Response File: Author Response.pdf

Reviewer 4 Report

After carefully reading the manuscript entitled: "Use of unmalted and malted buckwheat in brewing" it can be concluded that the authors spent a lot of time and effort in conducting experiments and writing an article. The topic is very attractive. However, many things need to be worked through and explained. Below are remarks, clarification requests, and suggestions.

1.     Extensive editing of the English language and style is required.

2.     The sentences are too long, and very often, it is difficult to understand the meaning and hard to follow.

3.     Text marked in yellow and red should be corrected and/or checked for data accuracy.

4.     It is necessary to restructure the entire text and separate the methods from the results and conclusions. In the current version, conclusions can be found in the material and method of work. In the results, we can find material and method, etc...

5.     The method of beer production is not clearly defined. The procedures in the material and method are not clear. The procedure described in the material and method differs from the one described in the results.

6.     Quite a lot of results are contradictory and not following literature data. Moreover, it is not in line with the results presented at the beginning of the article.

7.     Self-plagiarism is present to a certain extent. The article: Buckwheat and Amaranth as Raw Materials for Brewing, a Review in Plants contains identical sentence parts. Also, identical sentences can be found in some other articles written by the same authors.

8.     There are already published results in other journals by the same authors listed here as a new one. The article: Comparative evaluation of the physico-chemical characteristics of buckwheat malt and barley malt published in December 2021 have identical data in tables. Some parts of the text are identical, table 1 and 3, parts of table 2, etc. Furthermore, Research on obtaining some types of beer from unconventional raw materials contain already published results.

9.     The graphical representations (Fig.1-Fig.3). they are not clear, some correlations are contradictory, and their purpose is unclear. There is a possibility that it will confuse the reader more than help them understand.

10.  Some cited references are not the latest, even though the text states otherwise. Sample reference 26. Analytica EBC, 1998. European Brewery Convention. Verlag Hans Carl Getr¨anke-Fachverlag, Nürnberg, Germany

Based on all of the above, the paper needs major revision to be published in the journal Applied Sciences (ISSN 2076-3417). it is in the best interest of the author and the journal

Comments for author File: Comments.zip

Author Response

Response to Reviewer 4

Dear Referee,  

We would like to thank the referee for the close reading and for the proper suggestions.

We hope that we provide all the answers to the reviewer’s comments.

Thank you very much for the recommendations to publish our paper entitled “Use of unmalted and malted buckwheat in brewing”.

The present version of the paper has been revised according to the reviewer’s suggestions.       

We uploaded the corrected version of the article for which we used the green colour for the addition text.

Kindly take notice of these changes made to the manuscript's text. The green color indicates the answer to your suggestions, while the red and blue colours indicate the responses to the recommendations of the other three reviewers. We regret not include the responses to your suggestions in this letter. The paper (V2) has been attached with all the revisions made per the suggestions of the four reviewers.

Point 1: Extensive editing of the English language and style is required.

Response 1: First of all, we would like to thank the referee for the close reading and for all the given comments suitable for improving the manuscript. We made the English language corrections throughout the manuscript (in green).

Point 2: The sentences are too long, and very often, it is difficult to understand the meaning and hard to follow.

Response 2: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions.

Point 3: Text marked in yellow and red should be corrected and/or checked for data accuracy.

Response 3: We would like to thank to the referee for her/his remarks. We checked and corrected the entire manuscript.

For example, Table 3 (Table 1):

The protein content of the raw materials is important for the nutrition of the yeast during the fermentation stage.  These are the values that were obtained through our laboratory determinations.

In the specialized literature, I found these intervals for the starch content of the two raw materials: “The availability of starch varies between 70–91%, an important aspect for the use of buckwheat as a raw material in the beer industry”, “ content in starch for barley (52.10–69.08%”. The values for the starch content of the two raw materials are the ones we found through the determinations made, respectively for barley, 62.00%, and for buckwheat, 67%.

Table 4 (Table 2). These are the values we found on the two types of malt, values that were compared and explained the differences with those in the specialized literature. Yes, indeed for buckwheat malt brewing the blend of amylases, endo-protease, beta-glucanases, and hemicellulasesenzymes have to be used. For these researches we only had this commercial enzyme preparation Termamyl classic at our disposal.

Point 4: It is necessary to restructure the entire text and separate the methods from the results and conclusions. In the current version, conclusions can be found in the material and method of work. In the results, we can find material and method, etc.

Response 4: We would like to thank to the referee for her/his remarks. We revised the entire manuscript according to your instructions.

Point 5: The method of beer production is not clearly defined. The procedures in the material and method are not clear. The procedure described in the material and method differs from the one described in the results.

Response 5: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions.

Point 6: Quite a lot of results are contradictory and not following literature data. Moreover, it is not in line with the results presented at the beginning of the article.

Response 6: We would like to thank to the referee for her/his remarks. Your recommendations were taken into consideration, and the results and discussions section was restructured.

Point 7: Self-plagiarism is present to a certain extent. The article: Buckwheat and Amaranth as Raw Materials for Brewing, a Review in Plants contains identical sentence parts. Also, identical sentences can be found in some other articles written by the same authors.

Response 7: We would like to thank to the referee for her/his remarks. We made this adjustment. We just mentioned that we had done study on buckwheat in the beer malt field; we left out the self-plagiarism part.

Point 8: There are already published results in other journals by the same authors listed here as a new one. The article: Comparative evaluation of the physico-chemical characteristics of buckwheat malt and barley malt published in December 2021 have identical data in tables. Some parts of the text are identical, table 1 and 3, parts of table 2, etc. Furthermore, Research on obtaining some types of beer from unconventional raw materials contain already published results.

Response 8: We would like to thank to the referee for her/his remarks. We made this adjustment. we left out the self-plagiarism part.

Point 9: The graphical representations (Fig.1-Fig.3). they are not clear, some correlations are contradictory, and their purpose is unclear. There is a possibility that it will confuse the reader more than help them understand.

Response 9: We would like to thank to the referee for her/his remarks. We made the changes according to the referee suggestions. We made an attempt to more clearly describe the statistical analysis of the data for the final beer.

Point 10: Some cited references are not the latest, even though the text states otherwise. Sample reference 26. Analytica EBC, 1998. European Brewery Convention. Verlag Hans Carl Getr¨anke-Fachverlag, Nürnberg, Germany

Response 10: We would like to thank to the referee for her/his remarks. To the methods section, we added all the new EBC references.

Round 2

Reviewer 4 Report

Dear all,

I am generally satisfied with the modifications which were done. In the pdf file, I have highlighted in yellow and red what has to be corrected. These corrections are minor.

Kind regards

Comments for author File: Comments.pdf

Author Response

Response to Reviewer 4

Dear Referee,  

We would like to thank the referee for the close reading and for the proper suggestions.

We hope that we provide all the answers to the reviewer’s comments.

Thank you very much for the recommendations to publish our paper entitled “Use of unmalted and malted buckwheat in brewing”.

The present version of the paper has been revised according to the reviewer’s suggestions.       

We uploaded the corrected version of the article for which we used the green colour for the addition text.

Kindly take notice of these changes made to the manuscript's text. The green color indicates the answer to your suggestions, while the red and blue colours indicate the responses to the recommendations of the other three reviewers.

We regret not include the responses to your suggestions in this letter.

The paper (V3) has been attached with all the revisions made per the suggestions of the four reviewers.